METHODS FOR ALTERING BODY COMPOSITION BY ADMINISTERING A GDF8 INHIBITOR AND AN ACTIVIN A INHIBITOR

Abstract

The present invention relates to compositions and methods for altering body composition in a subject, wherein the alteration of body composition is an increase in muscle mass and a reduction of fat mass simultaneously. The present invention also relates to compositions and methods for reducing fat mass in a subject. The compositions and methods also increase muscle volume and lean body mass in the subject. The present invention also relates to compositions that comprise a GDF8 inhibitor and an Activin A inhibitor and the use of such compositions to treat diseases and disorders characterized by increased fat mass, and/or decreased muscle volume.

Description

CROSS REFERENCES TO RELATED APPLICATIONS

[0001] This application is a continuation of U.S. Non-Provisional application Ser. No. 16/290,287, filed 1 Mar. 2019, which claims the benefit of U.S. Provisional Application No. 62/637,017, filed 1 Mar. 2018, the entire contents of which are incorporated herein by reference.

SEQUENCE LISTING

[0002] This application includes a Sequence Listing in electronic format entitled "Sequence-Listing-40848-091USC1", which was created on 26 Feb. 2019 and which has a size of 288 kilobytes (KB) (295,202 bytes). The contents of txt file "Sequence-Listing-40848-091USC1" are incorporated by reference herein.

FIELD OF THE INVENTION

[0003] The present invention relates to compositions and methods for altering body composition in a subject. The compositions and methods also reduce fat mass in a subject. The compositions and methods also increase muscle volume and/or lean body mass in the subject. More specifically, the invention relates to compositions that comprise a GDF8 inhibitor and an Activin A inhibitor and the use of such compositions to treat diseases and disorders characterized by increased fat mass, and/or decreased muscle volume or decreased lean body mass.

BACKGROUND

[0004] Growth and differentiation factor-8 (GDF8, also known as myostatin), is a secreted ligand belonging to the transforming growth factor-.beta. (TGF-.beta.) superfamily of growth factors. GDF8 plays a central role in the development and maintenance of skeletal muscle, acting as a negative regulator of myogenesis and skeletal muscle mass. Myostatin mutations (including knockout) translate into phenotypes that are predominantly increased muscle mass, but can be made up of variations in muscling (more muscle fibers), muscle fiber composition (greater cross-sectional area of muscle fibers), increased protein/DNA ratio, and other.

[0005] Antibodies to GDF8 and therapeutic methods are disclosed in, e.g., U.S. Pat. No. 8,840,894. Anti-GDF8 antibodies are also mentioned in, e.g., U.S. Pat. Nos. 6,096,506; 7,320,789; 7,261,893; 7,807,159; 7,888,486; 7,635,760; 7,632,499; in US Patent Appl. Publ. Nos. 2006/0263354; 2007/0178095; 2008/0299126; 2010/0166764; 2009/0148436; and International Patent Appl. Publ. Nos. WO2004/037861; WO2007/047112; WO 2010/070094.

[0006] Activins belong to the transforming growth factor-beta (TGF-.beta.) superfamily and exert a broad range of biological effects on cell proliferation, differentiation, metabolism, homeostasis, and apoptosis, as well as immune response and tissue repair. Activin A is a disulfide-linked homodimer (two beta-A chains) that binds to and activates heteromeric complexes of a type I (Act RI-A and Act RI-B) and a type II (Act RII-A and Act RII-B) serine-threonine kinase receptor.

[0007] Antibodies to Activin A and uses thereof are disclosed in, e.g., U.S. Pat. Nos. 8,309,082; 9,718,881; and International Patent Appl. Publ. No. WO2008/031061.

[0008] Compositions comprising an anti-GDF8 antibody and an anti-Activin A antibody and therapeutic methods are disclosed in, e.g., U.S. Pat. No. 8,871,209.

[0009] Obesity is a global problem for over a third of the world population. In the United States of America, the average obesity rate is over 20%. The costs of obesity-related illness are staggering, amounting to $190.2 billion, roughly 21% of annual medical costs in the U.S. Obesity is an epidemic disease characterized by chronic low-grade inflammation associated with dysfunctional (elevated) fat mass. In Framingham Heart Study participants, abdominal adiposity was associated with incident cardiovascular disease (CVD) after adjustments for clinical risk factors and overall adiposity. Britton JACC 2013 62; 921. Abdominal visceral fat accumulation was positively associated with the progression of coronary noncalcified plaque. Imai Atherosclerosis 2012. Because high fat mass is associated with such serious conditions as congestive heart failure, high blood pressure/hypertension, pulmonary embolism, osteoarthritis, lymphedema, gastro-esophageal reflux disease, chronic renal failure, cancer, fatty-liver disease, and even depression, there remains a need for therapies that reduce total fat and/or android fat mass in subjects.

BRIEF SUMMARY OF THE INVENTION

[0010] In one aspect, the invention is directed to a method for altering body composition in a subject, i.e., increase muscle mass and decreasing fat mass, comprising administering a first composition comprising an effective amount of a GDF8 inhibitor and a second composition comprising an effective amount of an Activin A inhibitor to the subject. In another aspect, the invention is directed to a method for inducing a reduction in fat mass in a subject comprising administering an effective amount of a composition comprising a GDF8 inhibitor and an Activin A inhibitor to the subject.

[0011] In another aspect, the invention is directed to a method for treating a disease or disorder characterized by or associated with increased fat mass, the method comprising administering to a subject in need thereof a first composition comprising an effective amount of a GDF8 inhibitor and a second composition comprising an effective amount of an Activin A inhibitor. In another aspect, the invention is directed to a method for treating a disease or disorder characterized by or associated with increased fat mass, the method comprising administering to a subject in need thereof an effective amount of a composition comprising a GDF8 inhibitor and an Activin A inhibitor. In one aspect, the invention is directed to a method for altering body composition in a subject, i.e., increase muscle mass and decreasing fat mass, comprising administering an effective amount of a GDF8 inhibitor and an effective amount of an Activin A inhibitor to the subject, wherein there is an insignificant change in total body mass. Thus, in one aspect of the invention, a subject administered an effective amount of a GDF8 inhibitor and an effective amount of an Activin A inhibitor will experience an increase in muscle mass concurrently with a decrease in fat mass, leading to a minimal and/or insignificant change in total body mass.

[0012] In one aspect, the invention is directed to a use of a GDF8 inhibitor and/or an Activin A inhibitor in the preparation of a medicament for achieving a reduction in fat mass in a subject. In another aspect, the invention is directed to a use of a GDF8 inhibitor and/or an Activin A inhibitor in the preparation of a medicament for treating a disease or disorder associated with increased fat mass in a subject.

[0013] In some embodiments, a GDF8 inhibitor is provided for use in a method for treating a disease or disorder characterized by increased fat mass, wherein the method comprises administrating to a subject the GDF8 inhibitor and an Activin A inhibitor.

[0014] In some embodiments, an Activin A inhibitor is provided for use in a method for treating a disease or disorder characterized by increased fat mass, wherein the method comprises administrating to a subject the Activin A inhibitor and a GDF8 inhibitor.

[0015] In some embodiments, a non-therapeutic method is provided for decreasing fat mass in a subject, the method comprising administering to the subject an Activin A inhibitor and a GDF8 inhibitor.

[0016] In one embodiment of a method according to the invention, the effective amount of a GDF8 inhibitor comprises a dosing regimen selected from the group consisting of at least 0.1 mg/kg to about 10 gm/kg, 1 mg/kg to about 1 gm/kg, and 10 mg/kg to 100 mg/kg. In a further embodiment of a method according to the invention, the effective amount of a GDF8 inhibitor comprises a dosing regimen selected from the group consisting of a single dose of about 0.01 to about 20 mg/kg body weight, about 0.1 to about 10 mg/kg body weight, and about 0.1 to about 5 mg/kg body weight.

[0017] In another embodiment of a method according to the invention, the effective amount of an Activin A inhibitor comprises a dosing regimen selected from the group consisting of at least 0.1 mg/kg to about 10 gm/kg, 1 mg/kg to about 1 gm/kg, and 10 mg/kg to 100 mg/kg. In a further embodiment of a method according to the invention, the effective amount of an Activin A inhibitor comprises a dosing regimen selected from the group consisting of a single dose of about 0.01 to about 20 mg/kg body weight, about 0.1 to about 10 mg/kg body weight, and about 0.1 to about 5 mg/kg body weight.

[0018] In one embodiment of a method according to the invention, the effective amount of a GDF8 inhibitor is 6 mg/kg body weight and the effective amount of an Activin A inhibitor is 3 mg/kg body weight. In one embodiment of a method according to the invention, the effective amount of a GDF8 inhibitor is 6 mg/kg body weight and the effective amount of an Activin A inhibitor is 10 mg/kg body weight.

[0019] In one embodiment of a method according to the invention, the first composition is formulated for intravenous, subcutaneous, or oral administration. In another embodiment of a method according to the invention, the second composition is formulated for intravenous, subcutaneous, or oral administration. In certain embodiments of a method according to the invention, the first and second compositions are administered concurrently or sequentially to the subject.

[0020] In one embodiment of a method according to the invention, the first and second compositions are combined into a third composition prior to administration. In a further embodiment, the third composition is formulated for intravenous, subcutaneous, or oral administration.

[0021] In one embodiment, a method according to the invention further comprises measuring total fat mass in the subject before administration. In another embodiment, a method according to the invention further comprises measuring total fat mass in the subject after administration, and administering the first and second composition until the subject has a reduction in total fat mass of at least 2% to 8%, 2.5% to 6%, 3% to 4%, or at least 2.0%, at least 2.5%, at least 3.0%, or at least 3.5%, or more.

[0022] In one embodiment, a method according to the invention further comprises measuring android fat mass in the subject before administration. In another embodiment, a method according to the invention further comprises measuring android fat mass in the subject after administration, and administering the first and second composition until the subject has a reduction in android fat mass of at least 2% to 8%, 2.5% to 6%, 3% to 4%, or at least 2.0%, at least 2.5%, at least 3.0%, or at least 3.5%, or more.

[0023] In one embodiment, a method according to the invention further comprises measuring subcutaneous adipose tissue volume in the subject before administration. In another embodiment, a method according to the invention further comprises measuring subcutaneous adipose tissue volume in the subject after administration, and administering the first and second composition until the subject has a reduction in android fat mass of at least 2% to 8%, 2.5% to 6%, 3% to 4%, or at least 2.0%, at least 2.5%, at least 3.0%, or at least 3.5%, or more.

[0024] In some embodiments, a method is provided comprising administering to a subject in need thereof an effective amount of a GDF8 inhibitor and an effective amount of a Activin A inhibitor, wherein the GDF8 inhibitor and the Activin A inhibitor are co-administered within 48 hours or less, 24 hours or less, 12 hours or less, 6 hours or less, 3 hours or less, or 1 hour or less.

[0025] In some embodiments, a method is provided comprising administering to a subject in need thereof an effective amount of a GDF8 inhibitor and an effective amount of a Activin A inhibitor, wherein the subject exhibits a decrease in total fat mass, android fat mass, and/or subcutaneous adipose tissue volume.

[0026] In some embodiments, a method is provided comprising administering to a subject in need thereof an effective amount of a GDF8 inhibitor and an effective amount of a Activin A inhibitor, wherein the subject exhibits a decrease in total fat mass, android fat mass, and/or subcutaneous adipose tissue volume after 4 weeks or more, or 8 weeks or more, following administration.

[0027] In some embodiments, a method is provided comprising administering to a subject in need thereof an effective amount of a GDF8 inhibitor and an effective amount of a Activin A inhibitor, wherein the subject exhibits a decrease in total fat mass, android fat mass, and/or subcutaneous adipose tissue volume, wherein the subject does not exhibit reduced thigh intramuscular adipose tissue volume.

[0028] In some embodiments, a method is provided comprising administering to a subject in need thereof an effective amount of a GDF8 inhibitor and an effective amount of a Activin A inhibitor, wherein the subject exhibits a decrease in total fat mass, android fat mass, and/or subcutaneous adipose tissue volume, wherein the subject does not exhibit reduced thigh intramuscular adipose tissue volume after 4 weeks or more, or 8 weeks or more, following administration.

[0029] In some embodiments, a kit is provided comprising a first container containing an effective amount of a GDF8 inhibitor and a second container containing an effective amount of a specific Activin A inhibitor.

[0030] In some embodiments, the GDF8 inhibitor is an isolated antibody or an antigen-binding fragment thereof that specifically binds to GDF8.

[0031] In one embodiment of a method according to the invention, the GDF8 inhibitor is an antibody or an antigen-binding fragment thereof that specifically binds to GDF8. In another embodiment, the antibody or antigen-binding fragment that specifically binds GDF8 comprises the heavy chain complementarity determining regions (HCDRs) of a heavy chain variable region (HCVR) comprising SEQ ID NO:360, and the light chain complementarity determining regions (LCDRs) of a light chain variable region (LCVR) comprising SEQ ID NO:368. In still another embodiment, the antibody or antigen-binding fragment that specifically binds GDF8 comprises three HCDRs comprising SEQ ID NO:362, SEQ ID NO:364, and SEQ ID NO:366, and three LCDRs comprising SEQ ID NO:370, SEQ ID NO:372, and SEQ ID NO:374.

[0032] In some embodiments, the Activin A inhibitor is an isolated antibody or an antigen-binding fragment thereof that specifically binds to Activin A.

[0033] In one embodiment of a method according to the invention, the Activin A inhibitor is an antibody or antigen-binding fragment thereof that specifically binds Activin A. In another embodiment, the antibody or antigen-binding fragment that specifically binds Activin A comprises the heavy chain complementarity determining regions (HCDRs) of a heavy chain variable region (HCVR) comprising SEQ ID NO:553, and the light chain complementarity determining regions (LCDRs) of a light chain variable region (LCVR) comprising SEQ ID NO:537. In still another embodiment, the antibody or antigen-binding fragment that specifically binds Activin A comprises three HCDRs comprising SEQ ID NO:555, SEQ ID NO:557, and SEQ ID NO:559, and three LCDRs comprising SEQ ID NO:539, SEQ ID NO:541, and SEQ ID NO:543.

[0034] In one embodiment of a method according to the invention, the effective dose of the Activin A inhibitor is selected from the group comprising between 100% to 200% of the effective dose of the GDF8 inhibitor, between 100% and 250% of the effective dose of the GDF8 inhibitor, between 100% and 300% of the effective dose of the GDF8 inhibitor, and between 100% and 400% by weight of the effective dose of the GDF8 inhibitor.

[0035] In another embodiment of a method according to the invention, the weight ratio of the effective dose of the Activin A inhibitor to effective dose of the GDF8 inhibitor is from 10:1 to 1:10, 8:1 to 1:8, 6:1 to 1:6, 3:1 to 1:3, or about 2:1 to 1:2. In another embodiment of a method according to the invention, the weight ratio of the effective dose of the Activin A inhibitor is about 1.5 to 2.0 times as large by weight as the amount the GDF8 inhibitor.

[0036] In one embodiment of a method according to the invention, the GDF8 inhibitor is a bispecific antibody or antigen-binding fragment thereof that specifically binds GDF8 and also specifically binds Activin A. In another embodiment, the Activin A inhibitor is a bispecific antibody or antigen-binding fragment thereof that specifically binds Activin A and also specifically binds GDF8.

[0037] In one embodiment of a method according to the invention, the reduction of fat mass in the subject is a reduction in total fat mass as measured by DXA (Dual-energy X-ray absorptiometry). In another embodiment of a method according to the invention, the reduction of fat mass in the subject is a reduction in android fat mass as measured by DXA (Dual-energy X-ray absorptiometry).

[0038] In one embodiment of a method according to the invention, the reduction of fat mass in the subject is a reduction in subcutaneous adipose tissue volume as measured by MRI (Magnetic Resonance Imaging).

[0039] In one embodiment of a method according to the invention, the subject experiences an increase in muscle volume. The muscle volume may be thigh muscle tissue volume, for example, as measured by MRI. In some embodiments, the muscle volume may be thigh muscle tissue volume, for example, as measured by MRI. In some embodiments, the thigh muscle volume may be thigh muscle tissue volume including intramuscular adipose tissue and large vessels, or thigh muscle tissue volume excluding intramuscular adipose tissue and large vessels, for example, as measured by MRI.

[0040] In one embodiment of a method according to the invention, the subject experiences an increase in total lean mass. The total lean mass may be measured by DXA (dual x-ray absorptiometry.

[0041] In one embodiment of a method according to the invention, the subject experiences an increase in appendicular lean body mass. The appendicular lean body mass may be measured by DXA, and, for example, calculated by aLBM equation.

[0042] In one embodiment of a method according to the invention, the subject experiences a decrease in total fat mass, for example, as measured by DXA.

[0043] In one embodiment of a method according to the invention, the subject experiences a decrease in android fat mass, for example, as measured by DXA.

[0044] In one embodiment of a method according to the invention, the subject experiences a decrease in subcutaneous adipose tissue volume, for example, as measured by DXA.

[0045] In one embodiment of a method according to the invention, the subject experiences a decrease in sum of fat mass of arms and legs, for example, as measured by DXA.

[0046] In one embodiment of the invention, the subject does not exhibit a decrease in thigh intramuscular adipose tissue volume, for example, as measured by MRI.

[0047] In one embodiment of the invention, the subject does not exhibit a decrease in total bone mineral density (BMD) mass, for example, as measured by DXA.

[0048] In one embodiment of the invention, the subject does not exhibit a decrease in total bone mineral content (BMC) mass, for example, as measured by DXA.

[0049] In one embodiment of the invention, the subject exhibits an increase in total bone mineral content (BMC) mass, for example, as measured by DXA.

[0050] In another embodiment of a method according to the invention, the subject does not have a muscle wasting condition or disease.

[0051] In some embodiments, a kit is provided for use in altering body composition, decreasing fat mass, increasing lean mass, or treating a disease or disorder characterized by or associated with increased fat mass, the kit comprising a first container comprising a composition comprising an effective amount of a GDF8 inhibitor and a second container comprising a second composition comprising an effective amount of an Activin A inhibitor.

[0052] In some embodiments, a GDF8 inhibitor is provided for use in manufacture of a first composition for use as a medicament in a kit for altering body composition, decreasing fat mass, increasing lean mass, or treating a disease or disorder characterized by or associated with increased fat mass, the kit further comprising a second composition comprising an Activin A inhibitor.

[0053] In some embodiments, an Activin A inhibitor is provided for use in manufacture of a first composition for use as a medicament in a kit for altering body composition, decreasing fat mass, increasing lean mass, or treating a disease or disorder characterized by or associated with increased fat mass, the kit further comprising a second composition comprising a GDF8 inhibitor.

[0054] In some embodiments, a GDF8 inhibitor is provided for use in manufacture of a first composition for use in a kit for altering body composition, decreasing fat mass, or increasing lean mass in a subject, the kit further comprising a second composition comprising an Activin A inhibitor.

[0055] In some embodiments, an Activin A inhibitor is provided for use in manufacture of a first composition for use in a kit for altering body composition, decreasing fat mass, or increasing lean mass in a subject, the kit further comprising a second composition comprising a GDF8 inhibitor.

[0056] In some embodiments, a first composition comprising a GDF8 inhibitor is provided for use in altering body composition, decreasing fat mass, increasing lean mass, or treating a disease or disorder characterized by or associated with increased fat mass or decreased lean mass in a subject, wherein the subject has received a second composition comprising an Activin A inhibitor.

[0057] In some embodiments, a first composition comprising an Activin A inhibitor is provided for use in altering body composition, decreasing fat mass, increasing lean mass, or treating a disease or disorder characterized by or associated with increased fat mass or decreased lean mass in a subject, wherein the subject has received a second composition comprising a GDF8 inhibitor.

[0058] In some embodiments, a first composition comprising a GDF8 inhibitor is provided for use in method for altering body composition, decreasing fat mass, increasing lean mass, or treating a disease or disorder characterized by or associated with increased fat mass or decreased lean mass, the method further comprising administering a second composition comprising an Activin A inhibitor.

[0059] In some embodiments, a first composition comprising an Activin A inhibitor is provided for use in method for altering body composition, decreasing fat mass, increasing lean mass, or treating a disease or disorder characterized by or associated with increased fat mass or decreased lean mass, the method further comprising administering a second composition comprising an GDF8 inhibitor.

[0060] In some embodiments, a composition is provided comprising an Activin A inhibitor and a GDF8 inhibitor for use in altering body composition, decreasing fat mass, increasing lean mass, or treating or preventing a disease or disorder characterized by or associated with increased fat mass or decreased lean mass.

[0061] Other embodiments of the present invention will become apparent from a review of the ensuing detailed description.

BRIEF DESCRIPTION OF THE FIGURES

[0062] FIG. 1 shows a bar graph depicting results of clinical studies in sarcopenia patients 70 years of age or older receiving anti-GDF8 antibody REGN1033 alone after 12 weeks as the percent change from baseline of total lean mass in LS mean and SE compared to placebo. Patients receiving REGN1033 exhibited significantly increased total lean body mass at 12 weeks at each of three dosing regimens when compared to placebo (n=65). Patients receiving 100 mg anti-GDF8 antibody REGN1033 Q4W S.C. exhibited a difference vs. placebo of 1.66% total lean mass (n=62, P=0.0077). Patients receiving 300 mg anti-GDF8 Q4W SC exhibited a difference vs. placebo of 1.78% total lean mass (n=64, P=0.0043). Patients receiving 300 mg Q2W SC exhibited a difference vs. placebo of 2.29% total lean mass (n=59, P=0.0004).

[0063] FIG. 2A shows Table 1 with the ascending dose panels used in the study of 48 healthy post-menopausal women according to Example 2. A single intravenous dose of anti-GDF8 antibody REGN1033 and/or anti-Activin A antibody REGN2477 was utilized. In the primary analyses, the placebo and high dose combination groups were pooled across panels, yielding 12 subjects on placebo and 12 on the high dose combination, as shown by boxed regions in the table.

[0064] FIG. 2B shows a bar graph depicting thigh muscle volume (measured via MRI) % change at week 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women. A significant increase in thigh muscle volume % change from placebo was exhibited by anti-GDF8 (6 mg/kg), anti-GDF8 (6 mg/kg)+mid dose anti-Activin A (3 mg/kg), and anti-GDF8 (6 mg/kg)+high dose anti-Activin A (10 mg/kg) groups. (*nominal p<0.5 vs. placebo, **** nominal p<0.0001 vs. placebo).

[0065] FIG. 2C shows a bar graph depicting total fat mass (measured via DXA) % change at week 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women. The numbers show changes from placebo. A significant decrease in total fat mass % change was exhibited by the anti-GDF8 (6 mg/kg)+high dose anti-Activin A (10 mg/kg) group. (*nominal p<0.05 vs. placebo).

[0066] FIG. 3 shows a line graph depicting LS mean (SE) percent change from baseline in thigh muscle volume by MRI at weeks 0, 4, and 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women. At 4 weeks after single I.V. dose, a significant increase in thigh muscle volume (measured by MRI excluding intramuscular adipose tissue) % change from placebo was exhibited by each of anti-GDF8 (6 mg/kg), anti-GDF8 (6 mg/kg)+low dose anti-Activin A (1 mg/kg), anti-GDF8 (6 mg/kg)+mid dose anti-Activin A (3 mg/kg), and anti-GDF8 (6 mg/kg)+high dose anti-Activin A (10 mg/kg) groups. At 8 weeks after single I.V. dose, a significant increase in thigh muscle volume % change from placebo was exhibited in anti-GDF8 (6 mg/kg)+mid dose anti-Activin A (3 mg/kg), and anti-GDF8 (6 mg/kg)+high dose anti-Activin A (10 mg/kg) groups. (*nominal p<0.05, **nominal p <0.001). N values for each group are shown in FIG. 2A.

[0067] FIG. 4 show line graphs depicting individual data for percent change (from baseline) in thigh muscle volume by MRI at weeks 0, 4, and 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women as % change from baseline in placebo, anti-GDF8 (6 mg/kg), high dose anti-Activin A (10 mg/kg), anti-GDF8(6 mg/kg)+low dose anti-Activin A(1 mg/kg), anti-GDF8(6 mg/kg)+mid dose anti-Activin A(3 mg/kg), and anti-GDF8(6 mg/kg)+high dose anti-Activin A (10 mg/kg) groups for multiple individuals. Increases in thigh muscle volume were consistently observed in individual subjects following treatment with REGN2477+REGN1033 in combination. Within each treatment group, different lines indicate different individuals.

[0068] FIG. 5 shows a line graph depicting appendicular lean (body) mass (i.e., sum of lean tissue in the arms and legs) at weeks 0, 4, and 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women as % percent change from baseline LS (least-squares) mean (SE) in six groups. N values for each group are shown in FIG. 2A. After 4 weeks and 8 weeks, each of the three combination dose groups, including anti-GDF8(6 mg/kg)+low dose anti-Activin A(1 mg/kg), anti-GDF8(6 mg/kg)+mid dose anti-Activin A(3 mg/kg), and anti-GDF8(6 mg/kg)+high dose anti-Activin A (10 mg/kg) groups, exhibited significantly increased % change LS mean difference in appendicular lean mass compared to placebo (* nominal p<0.05, ** nominal p<0.001). N values for each group are shown in FIG. 2A.

[0069] FIG. 6 shows a line graph depicting mean (SE=standard errors) total fat mass percent (as measured by DXA) change at weeks 0, 4, and 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women in placebo, anti-GDF8 (6 mg/kg), high dose anti-Activin A (10 mg/kg), anti-GDF8(6 mg/kg)+low dose anti-Activin A(1 mg/kg), anti-GDF8(6 mg/kg)+mid dose anti-Activin A(3 mg/kg), and anti-GDF8(6 mg/kg)+high dose anti-Activin A (10 mg/kg) groups. N values for each group are shown in FIG. 2A. The high dose group, (anti-GDF8(6 mg/kg)+high dose anti-Activin A (10 mg/kg), exhibited a significant reduction in total fat mass as percent change LS Mean difference compared to placebo at week 4 and week 8 (*nominal p<0.05). Blockade of both Activin A and GDF8 led to reductions in total fat mass, as assessed by DXA.

[0070] FIG. 7 shows a line graph depicting mean (SE) percent change in android fat mass at weeks 0, 4, and 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women in six groups including placebo, anti-GDF8(6 mg/kg), high dose (10 mg/kg) anti-Activin A, anti-GDF8(6 mg/kg)+low dose (1 mg/kg) anti-Activin A, anti-GDF8(6 mg/kg)+mid dose (3 mg/kg) anti-Activin A, and anti-GDF8 (6 mg/kg)+high dose (10 mg/kg) anti-Activin A groups. The high dose REGN1033+REGN2477 group exhibited significantly reduced percent change LS Mean difference in android fat mass by DXA compared to placebo at week 4 and week 8 (*nominal p<0.05). N values for each group are shown in FIG. 2A. Blockade of both Activin A and GDF8 was also associated with decreases in android fat mass, as assessed by DXA.

[0071] FIG. 8 shows a line graph depicting LS mean percent change with SE in thigh muscle volume (excluding intramuscular adipose tissue and large vessels) at weeks 0, 4, and 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women in six groups including placebo, anti-GDF8(6 mg/kg), high dose (10 mg/kg) anti-Activin A, anti-GDF8 (6 mg/kg)+low dose (1 mg/kg) anti-Activin A, anti-GDF8(6 mg/kg)+mid dose (3 mg/kg) anti-Activin A, and anti-GDF8(6 mg/kg)+high dose (10 mg/kg) anti-Activin A groups. N values for each group are shown in FIG. 2A. Compared with placebo, REGN2477+REGN1033 medium and high groups exhibited significantly increased mean % change in thigh muscle volume at 4 weeks and 8 weeks. (*nominal p<0.05, **nominal p<0.001).

[0072] FIG. 9 shows a line graph depicting LS mean percent change with SE in total lean mass at weeks 0, 4, and 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women for six groups including placebo, anti-GDF8(6 mg/kg), high dose (10 mg/kg) anti-Activin A, anti-GDF8 (6 mg/kg)+low dose (1 mg/kg) anti-Activin A, anti-GDF8 (6 mg/kg)+mid dose (3 mg/kg) anti-Activin A, and anti-GDF8(6 mg/kg)+high dose (10 mg/kg) anti-Activin A groups. N values for each group are shown in FIG. 2A. Compared with placebo, REGN2477+REGN1033 medium and high dose groups exhibited significantly increased % change in total lean mass compared to placebo at 4 weeks and 8 weeks. (*nominal p<0.05).

[0073] FIG. 10 shows a line graph depicting LS mean percent change with SE in appendicular lean body mass (calculated via aLBM equation), in kg, at weeks 0, 4, and 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women in six groups including placebo, anti-GDF8, high dose (10 mg/kg) anti-Activin A, anti-GDF8+low dose (1 mg/kg) anti-Activin A, anti-GDF8+mid dose (3 mg/kg) anti-Activin A, and anti-GDF8+high dose (10 mg/kg) anti-Activin A groups. N values for each group are shown in FIG. 2A. REGN2477+REGN1033 treatment resulted in significantly increased % change in appendicular lean body mass calculated by aLBM equation)(kg) at each low, medium and high dose groups at 4 weeks and 8 weeks compared with placebo (* p<0.05, ** p<0.001).

[0074] FIG. 11 shows a line graph depicting LS mean percent change with SE in total fat mass, in kg, at weeks 0, 4, and 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women in six groups including placebo, anti-GDF8 (6 mg/kg), high dose (10 mg/kg) anti-Activin A, anti-GDF8+low dose (1 mg/kg) anti-Activin A, anti-GDF8(6 mg/kg)+mid dose (3 mg/kg) anti-Activin A, and anti-GDF8 (6 mg/kg)+high dose (10 mg/kg) anti-Activin A groups. N values for each group are shown in FIG. 2A. The high dose combination REGN2477+REGN1033 treatment group exhibited significantly decreased % total fat mass: -3.92% (high dose group) compared with placebo -0.65% at 8 weeks. (*nominal p<0.05).

[0075] FIG. 12 shows a line graph depicting LS mean percent change with SE in thigh muscle volume, in cm.sup.3, (including intramuscular adipose tissue and large vessels) at weeks 0, 4, and 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women in six groups including placebo, anti-GDF8, high dose (10 mg/kg) anti-Activin A, anti-GDF8+low dose (1 mg/kg) anti-Activin A, anti-GDF8+mid dose (3 mg/kg) anti-Activin A, and anti-GDF8+high dose (10 mg/kg) anti-Activin A groups. N values for each group are shown in FIG. 2A. The high and medium dose REGN2477+REGN1033 treatment groups exhibited significantly increased % change in thigh muscle volume, including intramuscular adipose tissue and large vessels in medium and high treatment groups at 4 weeks and 8 weeks compared to placebo (*nominal p<0.05, **nominal p<0.001).

[0076] FIG. 13 shows a line graph depicting LS mean percent change with SE in appendicular lean mass (sum of lean mass of arms and legs), in kg, at weeks 0, 4, and 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women in six groups including placebo, anti-GDF8, high dose (10 mg/kg) anti-Activin A, anti-GDF8+low dose (1 mg/kg) anti-Activin A, anti-GDF8+mid dose (3 mg/kg) anti-Activin A, and anti-GDF8+high dose (10 mg/kg) anti-Activin A groups. N values for each group are shown in FIG. 2A. Appendicular lean mass was significantly increased as mean percent change from placebo in each REGN2477+REGN1033 low, medium and high treatment groups at 4 and 8 weeks (*nominal p<0.05, *nominal* p<0.001).

[0077] FIG. 14 shows a line graph depicting LS mean percent change with SE in android fat mass, in kg, at weeks 0, 4, and 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women in six groups including placebo, anti-GDF8, high dose (10 mg/kg) anti-Activin A, anti-GDF8+low dose (1 mg/kg) anti-Activin A, anti-GDF8+mid dose (3 mg/kg) anti-Activin A, and anti-GDF8+high dose (10 mg/kg) anti-Activin A groups. N values for each group are shown in FIG. 2A. The high dose REGN1033+REGN2477 group exhibited significantly reduced % change in android fat mass by DXA compared to placebo at week 4 and week 8 (*nominal p<0.05).

[0078] FIG. 15 shows a line graph depicting LS mean percent change with SE in thigh intramuscular adipose tissue volume, in cm.sup.3, at weeks 0, 4, and 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women in six groups including placebo, anti-GDF8, high dose (10 mg/kg) anti-Activin A, anti-GDF8+low dose (1 mg/kg) anti-Activin A, anti-GDF8+mid dose (3 mg/kg) anti-Activin A, and anti-GDF8+high dose (10 mg/kg) anti-Activin A groups. N values for each group are shown in FIG. 2A. The high dose REGN1033+REGN2477 group exhibited increased thigh intramuscular adipose tissue volume as mean % change compared to placebo at 8 weeks. (*nominal p<0.05).

[0079] FIG. 16 shows a line graph depicting LS mean percent change with SE in sum of intramuscular and perimuscular adipose tissue (IMAT) at weeks 0, 4, and 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women in six groups including placebo, anti-GDF8, high dose (10 mg/kg) anti-Activin A, anti-GDF8+low dose (1 mg/kg) anti-Activin A, anti-GDF8+mid dose (3 mg/kg) anti-Activin A, and anti-GDF8+high dose (10 mg/kg) anti-Activin A groups. N values for each group are shown in FIG. 2A. The combination REGN1033+REGN2477 treatment groups were not significantly different than placebo in LS mean percent change with SE in sum of intramuscular and perimuscular adipose tissue (IMAT) at weeks 4 and 8.

[0080] FIG. 17 shows a line graph depicting LS mean percent change with SE in subcutaneous adipose tissue volume, in cm.sup.3, at weeks 0, 4, and 8 after a single I.V. dose of anti-Activin A antibody REGN2477 and/or anti-GDF8 antibody REGN1033 in healthy post-menopausal women in six groups including placebo, anti-GDF8, high dose (10 mg/kg) anti-Activin A, anti-GDF8+low dose (1 mg/kg) anti-Activin A, anti-GDF8+mid dose (3 mg/kg) anti-Activin A, and anti-GDF8+high dose (10 mg/kg) anti-Activin A groups. N values for each group are shown in FIG. 2A. The REGN1033+REGN2477 low and medium treatment groups exhibited significantly decreased % change in subcutaneous adipose tissue volume compared to placebo at 8 weeks (* nominal p<0.05).

DETAILED DESCRIPTION

[0081] Before the present invention is described, it is to be understood that this invention is not limited to particular compositions, methods, and experimental conditions described, as such compositions, methods, and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0082] As used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, a reference to "a method" includes one or more methods, and/or steps of the type described herein and/or which will become apparent to those persons skilled in the art upon reading this disclosure.

[0083] As used herein, the term "about," when used in reference to a particular recited numerical value, means that the value may vary from the recited value by no more than 1%. For example, as used herein, the expression "about 100" includes 99 and 101 and all values in between (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0084] Although any methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, the preferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference in their entirety.

Antibodies and Antigen-Binding Fragments of Antibodies

[0085] The term "antibody", as used herein, is intended to refer to immunoglobulin molecules comprising four polypeptide chains, two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, as well as multimers thereof (e.g., IgM). Each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or V.sub.H) and a heavy chain constant region. The heavy chain constant region comprises three domains, C.sub.H1, C.sub.H2 and C.sub.H3. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or V.sub.L) and a light chain constant region. The light chain constant region comprises one domain (C.sub.L1). The V.sub.H and V.sub.L regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR). Each V.sub.H and V.sub.L is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In different embodiments of the invention, the FRs of the antibodies (or antigen-binding portion thereof) may be identical to the human germline sequences, or may be naturally or artificially modified. An amino acid consensus sequence may be defined based on a side-by-side analysis of two or more CDRs.

[0086] Antibodies may be referred to herein according to the following nomenclature: Fc prefix (e.g. "H1M," "H2aM," "H4H"), followed by a numerical identifier (e.g. "10446"), followed by a "P," "P2" or "N" suffix. Thus, according to this nomenclature, an antibody may be referred to herein as, e.g., "H4H10446P2". The H1M, H2M and H4H prefixes on the antibody designations used herein indicate the particular Fc region isotype of the antibody. For example, an "H2aM" antibody has a mouse IgG2a Fc, whereas an "H4H" antibody has a human IgG4 Fc. As will be appreciated by a person of ordinary skill in the art, an antibody having a particular Fc isotype can be converted to an antibody with a different Fc isotype (e.g., an antibody with a mouse IgG2a Fc can be converted to an antibody with a human IgG4, etc.), but in any event, the variable domains (including the CDRs) will remain the same, and the binding properties are expected to be identical or substantially similar regardless of the nature of the Fc domain.

[0087] The term "antibody," as used herein, also includes antigen-binding fragments of full antibody molecules. The terms "antigen-binding portion" of an antibody, "antigen-binding fragment" of an antibody, and the like, as used herein, include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds an antigen to form a complex. Antigen-binding fragments of an antibody may be derived, e.g., from full antibody molecules using any suitable standard techniques such as proteolytic digestion or recombinant genetic engineering techniques involving the manipulation and expression of DNA encoding antibody variable and optionally constant domains. Such DNA is known and/or is readily available from, e.g., commercial sources, DNA libraries (including, e.g., phage-antibody libraries), or can be synthesized. The DNA may be sequenced and manipulated chemically or by using molecular biology techniques, for example, to arrange one or more variable and/or constant domains into a suitable configuration, or to introduce codons, create cysteine residues, modify, add or delete amino acids, etc.

[0088] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units consisting of the amino acid residues that mimic the hypervariable region of an antibody (e.g., an isolated complementarity determining region (CDR) such as a CDR3 peptide), or a constrained FR3-CDR3-FR4 peptide. Other engineered molecules, such as domain-specific antibodies, single domain antibodies, domain-deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g. monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains, are also encompassed within the expression "antigen-binding fragment," as used herein.

[0089] An antigen-binding fragment of an antibody will typically comprise at least one variable domain. The variable domain may be of any size or amino acid composition and will generally comprise at least one CDR that is adjacent to or in frame with one or more framework sequences. In antigen-binding fragments having a V.sub.H domain associated with a V.sub.L domain, the V.sub.H and V.sub.L domains may be situated relative to one another in any suitable arrangement. For example, the variable region may be dimeric and contain V.sub.H-V.sub.H, V.sub.H-V.sub.L or V.sub.L-V.sub.L dimers. Alternatively, the antigen-binding fragment of an antibody may contain a monomeric V.sub.H or V.sub.L domain.

[0090] In certain embodiments, an antigen-binding fragment of an antibody may contain at least one variable domain covalently linked to at least one constant domain. Non-limiting, exemplary configurations of variable and constant domains that may be found within an antigen-binding fragment of an antibody used in the present invention include: (i) V.sub.H-C.sub.H1; (ii) V.sub.H-C.sub.H2; (iii) V.sub.H-C.sub.H3; (iv) V.sub.H-C.sub.H1-C.sub.H2; (v) V.sub.H-C.sub.H1-C.sub.H2-C.sub.H3; (vi) V.sub.H-C.sub.H2-C.sub.H3; (vii) V.sub.H-C.sub.L; (viii) V.sub.L-C.sub.H1; (ix) V.sub.L-C.sub.H2; (x) V.sub.L-C.sub.H3; (xi) V.sub.L-C.sub.H1-C.sub.H2; (xii) V.sub.L-C.sub.H1-C.sub.H2-C.sub.H3; (xiii) V.sub.L-C.sub.H2-C.sub.H3; and (xiv) V.sub.L-C.sub.L. In any configuration of variable and constant domains, including any of the exemplary configurations listed above, the variable and constant domains may be either directly linked to one another or may be linked by a full or partial hinge or linker region. A hinge region may consist of at least 2 (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids which result in a flexible or semi-flexible linkage between adjacent variable and/or constant domains in a single polypeptide molecule. Moreover, an antigen-binding fragment of an antibody used in the present invention may comprise a homo-dimer or hetero-dimer (or other multimer) of any of the variable and constant domain configurations listed above in non-covalent association with one another and/or with one or more monomeric V.sub.H or V.sub.L domain (e.g., by disulfide bond(s)).

[0091] Full antibody molecules and antigen-binding fragments may be monospecific or multispecific (e.g., bispecific). A multispecific antibody or antigen-binding fragment of an antibody will typically comprise at least two different variable domains, wherein each variable domain is capable of specifically binding to a separate antigen or to a different epitope on the same antigen. Any multispecific antibody format may be adapted for use in the context of an antibody or antigen-binding fragment described herein using routine techniques available in the art.

[0092] The antibodies used in the compositions and methods of the invention may function through complement-dependent cytotoxicity (CDC) or antibody-dependent cell-mediated cytotoxicity (ADCC). "Complement-dependent cytotoxicity" (CDC) refers to lysis of antigen-expressing cells by an antibody of the invention in the presence of complement. "Antibody-dependent cell-mediated cytotoxicity" (ADCC) refers to a cell-mediated reaction in which nonspecific cytotoxic cells that express Fc receptors (FcRs) (e.g., Natural Killer (NK) cells, neutrophils, and macrophages) recognize bound antibody on a target cell and thereby lead to lysis of the target cell. CDC and ADCC can be measured using assays that are well known and available in the art. (See, e.g., U.S. Pat. Nos. 5,500,362 and 5,821,337, and Clynes et al., Proc. Natl. Acad. Sci. (USA) 95:652-656 (1998)).

[0093] The antibodies described herein may comprise or consist of human antibodies and/or recombinant human antibodies, or fragments thereof. The term "human antibody", as used herein, includes antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Human antibodies may nonetheless include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo), for example in the CDRs and in particular CDR3. However, the term "human antibody", as used herein, is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.

[0094] The antibody molecules described herein may comprise or consist of recombinant human antibodies or antigen-binding fragments thereof. The term "recombinant human antibody", as used herein, is intended to include all human antibodies that are prepared, expressed, created or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell, antibodies isolated from a recombinant, combinatorial human antibody library, antibodies isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes (see e.g., Taylor et al. (1992) Nucl. Acids Res. 20:6287-6295) or antibodies prepared, expressed, created or isolated by any other means that involves splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. In certain embodiments, however, such recombinant human antibodies are subjected to in vitro mutagenesis (or, when an animal transgenic for human Ig sequences is used, in vivo somatic mutagenesis) and thus the amino acid sequences of the V.sub.H and V.sub.L regions of the recombinant antibodies are sequences that, while derived from and related to human germline V.sub.H and V.sub.L sequences, may not naturally exist within the human antibody germline repertoire in vivo. Recombinant antibodies may be used in an additional embodiment of the compositions and methods of the invention.

[0095] An "isolated antibody," as used herein, means an antibody that has been identified and separated and/or recovered from at least one component of its natural environment. For example, an antibody that has been separated or removed from at least one component of an organism, tissue or cell in which the antibody naturally exists or is naturally produced is an "isolated antibody" for purposes of the invention. An isolated antibody also includes an antibody in situ within a recombinant cell, as well as an antibody that has been subjected to at least one purification or isolation step. According to certain embodiments, an isolated antibody may be substantially free of other cellular material and/or chemicals. Isolated antibodies may be used in an additional embodiment of the compositions and methods of the invention.

[0096] A protein or polypeptide is "substantially pure," "substantially homogeneous" or "substantially purified" when at least about 60 to 75% of a sample exhibits a single species of polypeptide. The polypeptide or protein may be monomeric or multimeric. A substantially pure polypeptide or protein will typically comprise about 50%, 60, 70%, 80% or 90% w/w of a protein sample, usually about 95%, and preferably over 99% pure. Protein purity or homogeneity may be indicated by a number of means well known in the art, such as polyacrylamide gel electrophoresis of a protein sample, followed by visualizing a single polypeptide band upon staining the gel with a stain well known in the art. For certain purposes, higher resolution may be provided by using HPLC or other means well known in the art for purification.

[0097] The term "polypeptide analog or variant" as used herein refers to a polypeptide that is comprised of a segment of at least 25 amino acids that has substantial identity to a portion of an amino acid sequence and that has at least one of the following properties: (1) specific binding to GDF8 under suitable binding conditions, or (2) ability to block the biological activity of GDF8. Typically, polypeptide analogs or variants comprise a conservative amino acid substitution (or insertion or deletion) with respect to the naturally occurring sequence. Analogs typically are at least 20 amino acids long, at least 50, 60, 70, 80, 90, 100, 150 or 200 amino acids long or longer, and can often be as long as a full-length naturally-occurring polypeptide.

[0098] Preferred amino acid substitutions are those which: (1) reduce susceptibility to proteolysis, (2) reduce susceptibility to oxidation, (3) alter binding affinity for forming protein complexes, (4) alter binding affinities, and (4) confer or modify other physicochemical or functional properties of such analogs. Analogs can include various mutations of a sequence other than the naturally-occurring peptide sequence. For example, single or multiple amino acid substitutions (preferably conservative amino acid substitutions) may be made in the naturally occurring sequence (preferably in the portion of the polypeptide outside the domain(s) forming intermolecular contacts. A conservative amino acid substitution should not substantially change the structural characteristics of the parent sequence (e.g., a replacement amino acid should not tend to break a helix that occurs in the parent sequence, or disrupt other types of secondary structure that characterizes the parent sequence). Examples of art-recognized polypeptide secondary and tertiary structures are described in Proteins, Structures and Molecular Principles (Creighton 1984 W. H. Freeman and Company, New York; Introduction to Protein Structure (Branden & Tooze, eds., 1991, Garland Publishing, NY); and Thornton et at. 1991 Nature 354:105, which are each incorporated herein by reference.

[0099] Non-peptide analogs are commonly used in the pharmaceutical industry as drugs with properties analogous to those of the template peptide. These types of non-peptide compound are termed "peptide mimetics" or "peptidomimetics" (see, for example, Fauchere (1986) J. Adv. Drug Res. 15:29; and Evans et al. (1987) J. Med. Chem. 30:1229, which are incorporated herein by reference. Systematic substitution of one or more amino acids of a consensus sequence with a D-amino acid of the same type (e.g., D-lysine in place of L-lysine) may also be used to generate more stable peptides. In addition, constrained peptides comprising a consensus sequence or a substantially identical consensus sequence variation may be generated by methods known in the art (Rizo et al. (1992) Ann. Rev. Biochem. 61:387, incorporated herein by reference), for example, by adding internal cysteine residues capable of forming intramolecular disulfide bridges which cyclize the peptide.

[0100] As applied to polypeptides, the term "substantial identity" or "substantially identical" means that two peptide sequences, when optimally aligned, such as by the programs GAP or BESTFIT using default gap weights, share at least about 80% sequence identity, at least about 90%, at least about 95%, at least about 98% or at least about 99% sequence identity. Preferably, residue positions that are not identical differ by conservative amino acid substitutions. A "conservative amino acid substitution" is one in which an amino acid residue is substituted by another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). In general, a conservative amino acid substitution will not substantially change the functional properties of a protein. In cases where two or more amino acid sequences differ from each other by conservative substitutions, the percent sequence identity or degree of similarity may be adjusted upwards to correct for the conservative nature of the substitution. Means for making this adjustment are well-known to those of skill in the art. See, e.g., Pearson (1994) Methods Mol. Biol. 24:307-331, herein incorporated by reference. Examples of groups of amino acids that have side chains with similar chemical properties include 1) aliphatic side chains: glycine, alanine, valine, leucine and isoleucine; 2) aliphatic-hydroxyl side chains: serine and threonine; 3) amide-containing side chains: asparagine and glutamine; 4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; 5) basic side chains: lysine, arginine, and histidine; and 6) sulfur-containing side chains are cysteine and methionine. Preferred conservative amino acids substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamate-aspartate, and asparagine-glutamine. Alternatively, a conservative replacement is any change having a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al. (1992) Science 256:1443-45, herein incorporated by reference. A "moderately conservative" replacement is any change having a nonnegative value in the PAM250 log-likelihood matrix.

[0101] Sequence similarity for polypeptides, which is also referred to as sequence identity, is typically measured using sequence analysis software. Protein analysis software matches similar sequences using measures of similarity assigned to various substitutions, deletions and other modifications, including conservative amino acid substitutions. For instance, GCG contains programs such as "Gap" and "Bestfit" which can be used with default parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species of organisms or between a wild type protein and a mutein thereof. See, e.g., GCG Version 6.1. Polypeptide sequences also can be compared using FASTA using default or recommended parameters, a program in GCG Version 6.1. FASTA (e.g., FASTA2 and FASTA3) provides alignments and percent sequence identity of the regions of the best overlap between the query and search sequences (Pearson (2000), supra). Another preferred algorithm when comparing a sequence of the invention to a database containing a large number of sequences from different organisms is the computer program BLAST, especially blastp or tblastn, using default parameters. See, e.g., Altschul et al. (1990) J. Mol. Biol. 215:403-410 and Altschul et al. (1997) Nucleic Acids Res. 25:3389 402, each of which is herein incorporated by reference.

[0102] The length of polypeptide sequences compared for homology will generally be at least about 16 amino acid residues, at least about 20 residues, at least about 24 residues, at least about 28 residues, or at least about 35 residues. When searching a database containing sequences from a large number of different organisms, it is preferable to compare amino acid sequences.

[0103] In some embodiments, the invention relates to a method for altering the body composition of a subject comprising administering a first composition comprising an effective amount of a GDF8 inhibitor and a second composition comprising an effective amount of an Activin A inhibitor to the subject.

[0104] In some embodiments, the invention relates to a method for inducing a reduction in fat mass in a subject comprising administering a first composition comprising an effective amount of a GDF8 inhibitor and a second composition comprising an effective amount of an Activin A inhibitor to the subject.

[0105] In some embodiments, the invention relates to a method for inducing an increase in muscle mass in a subject, the method comprising administering to a subject in need thereof a first composition comprising an effective amount of a GDF8 inhibitor and a second composition comprising an effective amount of an Activin A inhibitor.

[0106] In some embodiments, the invention relates to a method for altering the body composition of a subject comprising administering a composition comprising an effective amount of a GDF8 inhibitor and an effective amount of an Activin A inhibitor to the subject.

[0107] In some embodiments, the invention relates to a method for inducing a reduction in fat mass in a subject comprising administering a composition comprising an effective amount of a GDF8 inhibitor and an effective amount of an Activin A inhibitor to the subject.

[0108] In some embodiments, the invention relates to a method for inducing an increase in muscle mass in a subject, the method comprising administering to a subject in need thereof a composition comprising an effective amount of a GDF8 inhibitor and an effective amount of an Activin A inhibitor.

[0109] In some embodiments, the subject is a human subject. The human subject may be an adult human subject. The subject may be a male or female subject. The subject may be a healthy subject. The subject may be suffering from unwanted fat mass. The subject may be suffering from, or at risk of suffering from, a disease and disorder characterized by increased fat mass, and/or decreased muscle volume or decreased lean body mass. The subject may be a post-menopausal female subject. The subject may be a male subject or female subject 40 years of age or older, 50 years of age or older, 60 years of age or older, or 70 years of age or older.

[0110] In some embodiments, the invention relates to a method comprising administering a composition comprising a GDF8 inhibitor and a composition comprising an Activin A inhibitor to a subject in need thereof for treating or preventing diseases and disorders characterized by increased fat mass, and/or decreased muscle volume or decreased lean body mass.

[0111] In some embodiments according to the invention, the subject has at least one disease or disorder that may be associated with increased fat mass. In some embodiments, the disease or disorder may be selected from the group consisting of obesity, metabolic syndromes, nutritional disorders, high cholesterol, dyslipidemia, cardiovascular disease, cellulitis, cancer (including of the colon, esophagus, kidney, pancreas, gallbladder, breast, or endometrium), polycystic ovarian syndrome, gout, gallbladder disease, sleep apnea, respiratory disorder, asthma, osteoarthritis, cataract, congestive heart failure, enlarged heart, high blood pressure/hypertension, pulmonary embolism, lymphedema, gastro-esophageal reflux disease, hernia, chronic renal failure, urinary incontinence, connective tissue diseases, and fatty-liver disease. In another embodiment, the disease or disorder may be sarcopenia.

GDF8 Inhibitors

[0112] The present invention includes methods for altering body composition, inducing a reduction in fat mass, and increasing lean mass in a subject, and methods for treating a disease or disorder characterized by increased fat mass in a subject, comprising administering a composition comprising an effective amount of a GDF8 inhibitor to the subject.

[0113] The term "GDF8" (also referred to as "growth and differentiation factor-8" and "myostatin") means the protein having the amino acid sequence of SEQ ID NO:340 (mature protein). According to the present invention, GDF8-specific binding proteins specifically bind GDF8 but do not bind other ActRIIB ligands such as GDF3, BMP2, BMP4, BMP7, BMP9, BMP10, GDF11, Activin A, Activin B, Activin AB, Nodal, etc.

[0114] As used herein, a "GDF8 inhibitor" is any agent that binds to or interacts with human GDF8 and interferes with or inhibits the normal biological function of GDF8 in vitro or in vivo. Non-limiting examples of categories of GDF8 inhibitors include small molecule GDF8 antagonists, nucleic acid-based inhibitors of GDF8 expression or activity (e.g., siRNA or antisense), peptide-based molecules that specifically interact with GDF8 (e.g., peptibodies), receptor molecules that specifically interact with GDF8, GDF8-binding scaffold molecules, proteins comprising a ligand-binding portion of a receptor that specifically binds GDF8, and anti-GDF8 aptamers or portions thereof. In a preferred embodiment, a GDF8 inhibitor that can be used in the context of the present invention is an anti-GDF8 antibody or antigen-binding fragment thereof that specifically binds human GDF8. Anti-GDF8 antibodies include neutralizing and/or blocking antibodies. The inhibition caused by anti-GDF8 neutralizing and/or blocking antibodies need not be complete, as long as it is detectable using appropriate assays.

[0115] As used herein, the expression "anti-GDF8 antibody" also includes multispecific antigen-binding molecules (e.g., bispecific antibodies), wherein at least one binding domain (e.g., "binding arm") of the multispecific antigen-binding molecule specifically binds GDF8.

[0116] Exemplary anti-GDF8 antibodies that can be used in the compositions and methods of the invention include, e.g., the fully-human anti-GDF8 antibody H4H1657N2, also known as REGN1033, (e.g., an anti-GDF8 antibody comprising the heavy and light chain variable regions having amino acid sequences SEQ ID NO: 360 and SEQ ID NO: 368, respectively, as set forth in U.S. Pat. No. 8,840,894). Other GDF8 antagonists that can be used in the compositions and methods of the invention include anti-GDF8 antibodies (e.g., the antibody designated 2_112_1, e.g., having ATCC deposit designation PTA-6574, or e.g., 2_112_K, e.g., having HCVR/LCVR amino acid sequences SEQ ID NOs: 620 and 621) as set forth in US 2006/0263354 and U.S. Pat. No. 7,807,159; anti-GDF8 antibodies (e.g., 12A5-5, e.g., having HCVR/LCVR amino acid sequences of SEQ ID NO: 622 and 623) as set forth in U.S. Pat. No. 8,999,343 and US Publication No. 2013/0209489; anti-GDF8 antibodies (e.g., 10B3H8L5, e.g., having HCVR/LCVR amino acid sequences of SEQ ID NO:624 and 625, and 10B3H8L5-Fc-disabled) as set forth in US Publication No. 2013/0142788; anti-GDF8 antibodies (e.g., stamulumab/MYO-29, e.g., having HCVR/LCVR amino acid sequences of SEQ ID NOs: 626 and 627) as set forth in U.S. Pat. Nos. 8,940,874 and 7,261,893; anti-GDF8 antibodies (e.g., RK22/PF-0625616, e.g., having HCVR/LCVR amino acid sequences of SEQ ID NO: 628 and 629) as set forth in U.S. Pat. No. 8,415,459; anti-GDF8 antibodies (e.g., JA-16, e.g., having CDRs of HCVR amino acid sequence of SEQ ID NO: 630) as set forth in U.S. Pat. No. 7,731,961; anti-GDF8 antibodies (e.g., RK35, e.g., having HCVR/LCVR amino acid sequences of SEQ ID NO: 631 and 632) as set forth in U.S. Pat. No. 8,496,934 or 7,888,486, anti-GDF8 antibodies (e.g., OGD1.0.0, e.g., having HCVR/LCVR amino acid sequences of SEQ ID NO: 633 and 634) as set forth in U.S. Pat. No. 8,992,913; anti-GDF8 Fab molecules as set forth in European Patent No. 1 773 041 B1, and anti-GDF8 antibodies (e.g., C12, e.g., having HCVR/LCVR amino acid sequences of SEQ ID NOs: 635 and 636, C12-N93H, and/or 510C2 having HCVR/LCVR amino acid sequences of SEQ ID NOs: 637 and 638) as set forth in, e.g., U.S. Pat. Nos. 7,635,760 and 8,063,188, anti-GDF8 antibodies (e.g. 41C1E4/landogrozumab/LY2495655, e.g., having HCVR/LCVR amino acid sequences of SEQ ID NO: 639 and 640) as set forth in U.S. Pat. No. 7,632,499. In some embodiments, the anti-GDF8 antibody may have the full length heavy chain and full length light chain amino acid sequences of landogrozumab, e.g., SEQ ID Nos 641 and 642, respectively. In some embodiments, the anti-GDF8 antibody may comprise three heavy chain CDRs (HCDRs) and three light chain CDRs (LCDRs) of landogrozumab, for example, by Chothia definition, according to SEQ ID Nos: 643/644/645/646/647/648, respectively.

[0117] In one embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (HCVR) having an amino acid sequence selected from the group consisting of SEQ ID NO:2, 18, 34, 50, 66, 82, 98, 114, 130, 146, 162, 178, 194, 210, 226, 242, 258, 274, 290, 306, 360, and 376, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity.

[0118] In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof comprises a light chain variable region (LCVR) amino acid sequence selected from the group consisting of SEQ ID NO:10, 26, 42, 58, 74, 90, 106, 122, 138, 154, 170, 186, 202, 218, 234, 250, 266, 282, 298, 314, 322, 368, and 384, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity.

[0119] In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof comprises a HCVR amino acid sequence and a LCVR amino acid sequence, wherein the HCVR/LCVR sequence pair is selected from the group consisting of SEQ ID NO:2/10, 18/26, 34/42, 50/58, 66/74, 82/90, 98/106, 114/122, 130/138, 146/154, 162/170, 178/186, 194/202, 210/218, 226/234, 242/250, 258/266, 274/282, 290/298, 306/314, 114/322, 360/368, and 376/384.

[0120] In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof comprises a HCVR amino acid sequence and a LCVR amino acid sequence, wherein the HCVR/LCVR sequence pair is selected from the group consisting of (HCVR/LCVR): 21-E5 (SEQ ID NO:34/42); 21-B9 (SEQ ID NO:18/26); 21-E9 (SEQ ID NO:98/106); 21-A2 (SEQ ID NO:2/10); 22-D3 (SEQ ID NO:50/58); 22-E6 (SEQ ID NO:66/74); 22-G10 (SEQ ID NO:82/90); 1A2 (SEQ ID NO:226/234); 20B12 (SEQ ID NO:274/282); 58C8 (SEQ ID NO:242/250); 19F2 (SEQ ID NO:258/266); 8D12-1 (SEQ ID NO:114/122); 4E3-7 (SEQ ID NO:194/202); 9B11-12 (SEQ ID NO:162/170); 4B9 (SEQ ID NO:226/234); 1H4-5 (SEQ ID NO:210/218); 9B4-3 (SEQ ID NO:178/186); 3E2-1 (SEQ ID NO:290/298); 4G3-25 (SEQ ID NO:306/314); 4B6-6 (SEQ ID NO:130/138); H4H1657N2 (SEQ ID NO:360/368); H4H1669P (SEQ ID NO:376/384).

[0121] In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof comprises a heavy chain complementarity determining region 3 (HCDR3) domain and a light chain CDR3 (LCDR3) domain, wherein the HCDR3 domain has an amino acid sequence selected from the group consisting of SEQ ID NO:8, 24, 40, 56, 72, 88, 104, 120, 136, 152, 168, 184, 200, 216, 232, 248, 264, 280, 296, 312, 366, and 382, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity, and the LCDR3 domain has an amino acid sequence selected from the group consisting of SEQ ID NO:16, 32, 48, 64, 80, 96, 112, 128, 144, 160, 176, 192, 208, 224, 240, 256, 272, 288, 304, 320, 328, 374, and 390, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity. In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof comprises an HCDR3/LCDR3 amino acid sequence pair selected from the group consisting of SEQ ID NO:8/16, 24/32, 40/48, 56/64, 72/80, 88/96, 104/112, 120/128, 136/144, 152/160, 168/176, 184/192, 200/208, 216/224, 232/240, 248/256, 264/272, 280/288, 296/304, 312/320, 120/328, 366/374, and 382/390.

[0122] In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof comprises heavy chain CDR1 (HCDR1) and CDR2 (HCDR2) domains and light chain CDR1 (LCDR1) and CDR2 (LCDR2) domains, wherein the HCDR1 domain has an amino acid sequence selected from the group consisting of SEQ ID NO:4, 20, 36, 52, 68, 84, 100, 116, 132, 148, 164, 180, 196, 212, 228, 244, 260, 276, 292, 308, 362, and 378, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity; the HCDR2 domain has an amino acid sequence selected from the group consisting of SEQ ID NO:6, 22, 38, 54, 70, 86, 102, 118, 134, 150, 166, 182, 198, 214, 230, 246, 262, 278, 294, 310, 364, and 380, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity; the LCDR1 domain has an amino acid sequence selected from the group consisting of SEQ ID NO:12, 28, 44, 60, 76, 92, 108, 124, 140, 156, 172, 188, 204, 220, 236, 252, 268, 284, 300, 316, 324, 370, and 386, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity and the LCDR2 domain has an amino acid sequence selected from the group consisting of SEQ ID NO:14, 30, 46, 62, 78, 94, 110, 126, 142, 158, 174, 190, 206, 222, 238, 254, 270, 286, 302, 318, 326, 372, and 388, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity. In another embodiment, the HCDR1, HCDR2 and HCDR3 domains have respective amino acid sequence combinations selected from the group consisting of SEQ ID NO:36/38/40, 116/118/120, 228/230/232, 362/364/366, and 378/380/382; and the LCDR1, LCDR2 and LCDR3 domains have respective amino acid sequence combinations selected from the group consisting of SEQ ID NO:44/46/48, 124/126/128, 236/238/240, 370/372/374, and 386/388/390.

[0123] In yet another embodiment, the heavy and light chain CDR domains of the anti-GDF8 antibody or antigen-binding fragment thereof (HCDR1/HCDR2/HCDR3/LCDR1/LCDR2/LCDR3) have amino acid sequence combinations selected from the group consisting of SEQ ID NO: 36/38/40/44/46/48 (e.g., 21-E5), 116/118/120/124/126/128 (e.g., 8D12), 228/230/232/236/238/240 (e.g., 1A2), 362/364/366/370/372/374 (e.g., H4H1657N2), and 378/380/382/386/388/390 (e.g., H4H1669P).

[0124] In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof comprises heavy and light chain CDR domains contained within heavy and light chain variable region (HCVR/LCVR) amino acid sequence pairs selected from the group consisting of SEQ ID NO: 2/10, 18/26, 34/42, 50/58, 66/74, 82/90, 98/106, 114/122, 130/138, 146/154, 162/170, 178/186, 194/202, 210/218, 226/234, 242/250, 258/266, 274/282, 290/298, 306/314, 114/322, 360/368, and 376/384.

[0125] Methods and techniques for identifying CDRs within HCVR and LCVR amino acid sequences are well known in the art and can be used to identify CDRs within the specified HCVR and/or LCVR amino acid sequences disclosed herein. Exemplary conventions that can be used to identify the boundaries of CDRs include, e.g., the Kabat definition, the Chothia definition, and the AbM definition. In general terms, the Kabat definition is based on sequence variability, the Chothia definition is based on the location of the structural loop regions, and the AbM definition is a compromise between the Kabat and Chothia approaches. See, e.g., Kabat, "Sequences of Proteins of Immunological Interest," National Institutes of Health, Bethesda, Md. (1991); Al-Lazikani et al., J. Mol. Biol. 273:927-948 (1997); and Martin et al., Proc. Natl. Acad. Sci. USA 86:9268-9272 (1989). Public databases are also available for identifying CDR sequences within an antibody.

[0126] In one embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof comprises a HCVR having an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, 17, 33, 49, 65, 81, 97, 113, 129, 145, 161, 177, 193, 209, 225, 241, 257, 273, 289, 305, 359, and 375, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity.

[0127] In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof comprises a LCVR having an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:9, 25, 41, 57, 73, 89, 105, 121, 137, 153, 169, 185, 201, 217, 233, 249, 265, 281, 297, 313, 321, 367, and 383, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity.

[0128] In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof comprises a HCVR/LCVR pair having amino acid sequences encoded by a nucleic acid molecule pair selected from the group consisting of SEQ ID NO: 1/9, 17/25, 33/41, 49/57, 65/73, 81/89, 97/105, 113/121, 129/137, 145/153, 161/169, 177/185, 193/201, 209/217, 225/233, 241/249, 257/265, 273/281, 289/297, 305/313, 113/321, 359/367, and 375/383.

[0129] In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof comprises a HCDR3 domain having an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:7, 23, 39, 55, 71, 87, 103, 119, 135, 151, 167, 183, 199, 215, 231, 247, 263, 279, 295, 311, 365, and 381, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity, and a LCDR3 domain having an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:15, 31, 47, 63, 79, 95, 111, 127, 143, 159, 175, 191, 207, 223, 239, 255, 271, 287, 303, 319, 327, 373, and 389, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity. In one embodiment, the HCDR3/LCDR3 domain pair has amino acid sequences encoded by a nucleic acid sequence pair selected from the group consisting of SEQ ID NO:7/15, 23/31, 39/47, 55/63, 71/79, 87/95, 103/111, 119/127, 135/143, 151/159, 167/175, 183/191, 199/207, 215/223, 231/239, 247/255, 263/271, 279/287, 295/303, 311/319, 119/327, 365/373, and 381/389.

[0130] In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof comprises HCDR1 and HCDR2 domains, and LCDR1 and LCDR2 domains, wherein the HCDR1 domain has an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:3, 19, 35, 51, 67, 83, 99, 115, 131, 147, 163, 179, 195, 211, 227, 243, 259, 275, 291, 307, 361, and 377, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity, the HCDR2 domain has an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:5, 21, 37, 53, 69, 85, 101, 117, 133, 149, 165, 181, 197, 213, 229, 245, 261, 277, 293, 309, 363, and 379, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity, the LCDR1 domain has an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:11, 27, 43, 59, 75, 91, 107, 123, 139, 155, 171, 187, 203, 219, 235, 251, 267, 283, 299, 315, 323, 369, and 385, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity, and the LCDR2 domain has an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:13, 29, 45, 61, 77, 93, 109, 125, 141, 157, 173, 189, 205, 221, 237, 253, 269, 285, 301, 317, 325, 371, and 387.

[0131] In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof comprises heavy and light chain CDR domains (HCDR1/HCDR2/HCDR3/LCDR1/LCDR2/LCDR3) having amino acid sequence combinations encoded by a nucleic acid sequence set of SEQ ID NO:35/37/39/43/45/47, 115/117/119/123/125/127, 227/229/231/235/237/239, 361/363/365/369/371/373, or 377/379/381/385/387/389.

[0132] In a preferred embodiment, the anti-GDF8 antibody or antigen-binding fragment thereof that specifically binds GDF8 comprises the HCDRs of a heavy chain variable region (HCVR) comprising SEQ ID NO:360 and the LCDRs of a light chain variable region (LCVR) comprising SEQ ID NO:368. In another embodiment, the anti-GDF8 antibody or antigen-binding fragment that specifically binds GDF8 comprises three HCDRs comprising SEQ ID NO:362, SEQ ID NO:364, and SEQ ID NO:366 and three LCDRs comprising SEQ ID NO:370, SEQ ID NO:372, and SEQ ID NO:374.

[0133] In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof is a fully human or humanized antibody or antibody fragment that binds GDF8 with an affinity (expressed as a dissociation constant, "KD") of about 1 nM or less, as measured by surface plasmon resonance assay (for example, BIACORE.TM.). In certain embodiments, the antibody of the invention exhibits a KD of about 700 pM or less; about 500 pM or less; about 320 pM or less; about 160 pM or less; about 100 pM or less; about 50 pM or less; about 10 pM or less; or about 5 pM or less.

[0134] In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof is a fully human or humanized monoclonal antibody (mAb) that specifically binds and inhibits human GDF8 and exhibits an IC50 of less than or equal to about 10 nM; about 5 nM or less; about 3 nM or less; about 2 nM or less; about 1 nM or less; about 500 pM or less; or about 200 pM or less, as measured by GDF8 inducible luciferase assay.

[0135] In one embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof has a modified glycosylation pattern. In some applications, modification to remove undesirable glycosylation sites may be useful, or an antibody lacking a fucose moiety present on the oligosaccharide chain, for example, to increase antibody dependent cellular cytotoxicity (ADCC) function (Shield et al. (2002) JBC 277:26733). In other applications, modification of a galactosylation can be made in order to modify complement dependent cytotoxicity (CDC).

[0136] In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof competes for specific binding to GDF8 with another antibody comprising a HCDR1/HCDR2/HCDR3/LCDR1/LCDR2/LCDR3 domain combination having amino acid sequences selected from the group consisting of SEQ ID NO:36/38/40/44/46/48, 116/118/120/124/126/128, 228/230/232/236/238/240, 362/364/366/370/372/374, or 378/380/382/386/388/390. In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof competes for specific binding to GDF8 with another antibody comprising a HCVR/LCVR amino acid sequence pair of SEQ ID NO:2/10, 18/26, 34/42, 50/58, 66/74, 82/90, 98/106, 114/122, 130/138, 146/154, 162/170, 178/186, 194/202, 210/218, 226/234, 242/250, 258/266, 274/282, 290/298, 306/314, 114/322, 360/368, or 376/384.

[0137] In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof recognizes the epitope on GDF8 that is recognized by another antibody comprising a HCDR1/HCDR2/HCDR3/LCDR1/LCDR2/LCDR3 domain combination having amino acid sequences selected from the group consisting of SEQ ID NO: 36/38/40/44/46/48, 116/118/120/124/126/128, 228/230/232/236/238/240, 362/364/366/370/372/374, or 378/380/382/386/388/390. In another embodiment, an anti-GDF8 antibody or antigen-binding fragment thereof recognizes the epitope on GDF8 that is recognized by another antibody comprising a HCVR/LCVR amino acid sequence pair of SEQ ID NO:2/10, 18/26, 34/42, 50/58, 66/74, 82/90, 98/106, 114/122, 130/138, 146/154, 162/170, 178/186, 194/202, 210/218, 226/234, 242/250, 258/266, 274/282, 290/298, 306/314, 114/322, 360/368, or 376/384.

Activin A Inhibitors

[0138] The present invention includes methods for altering body composition, inducing a reduction in fat mass, and/or increasing lean mass in a subject, and methods for treating a disease or disorder characterized by increased fat mass in a subject comprising administering a composition comprising an effective amount of an Activin A inhibitor to the subject.

[0139] As used herein, an "Activin A inhibitor" is any agent that binds to or interacts with human Activin A and interferes with or inhibits the normal biological function of Activin A in vitro or in vivo. Non-limiting examples of categories of Activin A inhibitors include small molecule Activin A antagonists, nucleic acid-based inhibitors of Activin A expression or activity (e.g., siRNA or antisense), peptide-based molecules that specifically interact with Activin A (e.g., peptibodies), receptor molecules that specifically interact with Activin A, Activin A-binding scaffold molecules, proteins comprising a ligand-binding portion of a receptor that specifically binds Activin A, and anti-Activin A aptamers or portions thereof. In a preferred embodiment, an Activin A inhibitor that can be used in the context of the present invention is an anti-Activin A antibody or antigen-binding fragment thereof that specifically binds human Activin A. Anti-Activin A antibodies include neutralizing and/or blocking antibodies. The inhibition caused by anti-Activin A neutralizing and/or blocking antibodies need not be complete, as long as it is detectable using appropriate assays.

[0140] Activins are homo- and hetero-dimeric molecules comprising .beta.A and/or .beta.B subunits. The .beta.A subunit has the amino acid sequence of SEQ ID NO:617 and the .beta.B subunit has the amino acid sequence of SEQ ID NO: 619. Activin A is a homodimer of two .beta.A subunits; Activin B is a homodimer of two .beta.B subunits; and Activin AB is a heterodimer of one .beta.A subunit and one .beta.B subunit. An anti-Activin A antibody or antigen-binding fragment thereof specifically binds the .beta.A subunit. Since the .beta.A subunit is found in both Activin A and Activin AB molecules, an "anti-Activin A antibody or antigen-binding fragment thereof" can specifically bind Activin A, as well as Activin AB (by virtue of its interaction with the .beta.A subunit). Therefore, an anti-Activin A antibody or antigen-binding fragment thereof specifically binds Activin A, or Activin A and Activin AB, but does not bind other ActRIIB ligands, such as Activin B, GDF3, GDF8, BMP2, BMP4, BMP7, BMP9, BMP10, GDF11, Nodal, etc.

[0141] In some embodiments, an anti-Activin A antibody or antigen-binding fragment thereof is employed as set forth in U.S. Pat. No. 9,718,881. Exemplary anti-Activin A antibodies that can be used in the compositions and methods of the invention include, e.g., the fully-human anti-Activin antibody H4H10446P2, also known as REGN2477, (e.g., an anti-Activin A antibody comprising the heavy and light chain variable regions having amino acid sequences SEQ ID NO: 162 and SEQ ID NO: 146, respectively, as set forth in U.S. Pat. No. 9,718,881).

[0142] Table 2 sets forth heavy and light chain variable region amino acid sequence pairs of selected anti-Activin A antibodies and their corresponding antibody identifiers that can be used in the compositions and methods of the invention. The corresponding nucleic acid sequence identifiers are set forth in Table 3.

TABLE-US-00001 TABLE 2 anti-Activin A Amino Acid Sequence Identifiers SEQ ID NOs: Antibody Designation HCVR HCDR1 HCDR2 HCDR3 LCVR LCDR1 LCDR2 LCDR3 H4H10423P 393 395 397 399 401 403 405 407 H4H10424P 409 411 413 415 417 419 421 423 H4H10426P 425 427 429 431 433 435 437 439 H4H10429P 441 443 445 447 449 451 453 455 H4H10430P 457 459 461 463 465 467 469 471 H4H10432P2 473 475 477 479 481 483 485 487 H4H10433P2 489 491 493 495 481 483 485 487 H4H10436P2 497 499 501 503 481 483 485 487 H4H10437P2 505 507 509 511 481 483 485 487 H4H10438P2 513 515 517 519 481 483 485 487 H4H10440P2 521 523 525 527 481 483 485 487 H4H10442P2 529 531 533 535 537 539 541 543 H4H10445P2 545 547 549 551 537 539 541 543 H4H10446P2 553 555 557 559 537 539 541 543 H4H10447P2 561 563 565 567 537 539 541 543 H4H10448P2 569 571 573 575 537 539 541 543 H4H10452P2 577 579 581 583 537 539 541 543 H4H10468P2 585 -587 589 591 537 539 541 543 H2aM10965N 593 595 597 599 601 603 605 607

TABLE-US-00002 TABLE 3 anti-Activin A Nucleic Acid Sequence Identifiers SEQ ID NOs: Antibody Designation HCVR HCDR1 HCDR2 HCDR3 LCVR LCDR1 LCDR2 LCDR3 H4H10423P 392 394 396 398 400 402 404 406 H4H10424P 408 410 412 414 416 418 420 422 H4H10426P 424 426 428 430 432 434 436 438 H4H10429P 440 441 444 446 448 450 452 454 H4H10430P 456 458 460 462 464 466 468 470 H4H10432P2 472 474 476 478 480 482 484 486 H4H10433P2 488 490 492 494 480 482 484 486 H4H10436P2 496 498 500 502 480 482 484 486 H4H10437P2 504 506 508 510 480 482 484 486 H4H10438P2 512 514 516 518 480 482 484 486 H4H10440P2 520 522 524 526 480 482 484 486 H4H10442P2 528 530 532 534 536 538 540 542 H4H10445P2 544 546 548 550 536 538 540 542 H4H10446P2 552 554 556 558 536 538 540 524 H4H10447P2 560 562 564 566 536 538 540 542 H4H10448P2 568 570 572 574 536 538 540 542 H4H10452P2 576 578 580 582 536 538 540 542 H4H10468P2 584 586 588 590 536 538 540 542 H2aM10965N 592 594 596 598 600 602 604 606

[0143] In one embodiment, an anti-Activin A antibody or antigen-binding fragment thereof comprises a heavy chain variable region (HCVR) having an amino acid sequence selected from the group consisting of SEQ ID NO: 393, 409, 425, 441, 457, 473, 489, 497, 505, 513, 521, 529, 545, 553, 561, 569, 577, 585, and 593, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity.

[0144] In another embodiment, an anti-Activin A antibody or antigen-binding fragment thereof comprises a light chain variable region (LCVR) having an amino acid sequence selected from the group consisting of SEQ ID NO: 401, 417, 433, 449, 465, 481, 537, and 601, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity.

[0145] In another embodiment, an anti-Activin A antibody or antigen-binding fragment thereof comprises a HCVR and LCVR (HCVR/LCVR) amino sequence pair selected from the group consisting of SEQ ID NO: 393/401, 409/417, 425/433, 441/449, 457/465, 473/481, 489/481, 497/481, 505/481, 513/481, 521/481, 529/537, 545/537, 553/537, 561/537, 569/537, 577/537, 585/537, and 593/601.

[0146] In another embodiment, an anti-Activin A antibody or antigen-binding fragment thereof comprises a heavy chain CDR3 (HCDR3) domain having an amino acid sequence selected from the group consisting of SEQ ID NO: 399, 415, 431, 447, 463, 479, 495, 503, 511, 519, 527, 535, 551, 559, 567, 575, 583, 591, and 599, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity; and a light chain CDR3 (LCDR3) domain having an amino acid sequence selected from the group consisting of SEQ ID NO: 407, 423, 439, 455, 471, 487, 543, and 607, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity.

[0147] In another embodiment, an anti-Activin A antibody or antigen-binding fragment thereof comprises a HCDR3/LCDR3 amino acid sequence pair selected from the group consisting of SEQ ID NO: 399/407, 415/423, 431/439, 447/455, 463/471, 479/487, 495/487, 503/487, 511/487, 519/487, 527/487, 535/543, 551/543, 559/543, 567/543, 575/543, 583/543, 591/543, and 599/607.

[0148] In another embodiment, an anti-Activin A antibody or antigen-binding fragment thereof comprises a heavy chain CDR1 (HCDR1) domain having an amino acid sequence selected from the group consisting of SEQ ID NO: 395, 411, 427, 443, 459, 475, 491, 499, 507, 515, 523, 531, 547, 555, 563, 571, 579, 587, and 595, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity; a heavy chain CDR2 (HCDR2) domain having an amino acid sequence selected from the group consisting of SEQ ID NO: 397, 413, 429, 445, 461, 477, 493, 501, 509, 517, 525, 533, 549, 557, 565, 573, 581, 589, and 597, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity; a light chain CDR1 (LCDR1) domain having an amino acid sequence selected from the group consisting of SEQ ID NO: 403, 419, 435, 451, 467, 483, 539, and 603, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity; and a light chain CDR2 (LCDR2) domain having an amino acid sequence selected from the group consisting of SEQ ID NO: 405, 421, 437, 453, 469, 485, 541, and 605, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity.

[0149] In another embodiment, an anti-Activin A antibody or antigen-binding fragment thereof comprises HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 domains, respectively, having the amino acid sequences selected from the group consisting of: SEQ ID NOs: 395-397-8-403-405-407; 411-413-415-419-421-423; 36-429-431-435-437-439; 443-445-447-451-453-455; 459-461-463-467-469-471; 475-477-479-483-485-487; 491-493-495-483-485-487; 499-501-503-483-485-487; 507-509-511-483-485-487; 515-517-519-483-485-487; 523-525-527-483-485-487; 531-533-535-539-541-543; 547-549-551-539-541-543; 555-557-559-539-541-543 (H4H10446P2); 563-565-567-539-541-543; 571-573-575-539-541-543; 579-581-583-539-541-543; 587-589-591-539-541-543; and 595-597-599-603-605-607.

[0150] In another embodiment, an anti-Activin A antibody or antigen-binding fragment thereof comprises the heavy and light chain CDR domains contained within heavy and light chain variable region (HCVR/LCVR) sequence pairs selected from the group consisting of SEQ ID NO: 393/401, 409/417, 425/433, 441/449, 457/465, 473/481, 489/481, 497/481, 505/481, 513/481, 521/481, 529/537, 545/537, 553/537, 561/537, 569/537, 577/537, 585/537, and 593/601.

[0151] In one embodiment, an anti-Activin A antibody or antigen-binding fragment thereof comprises a HCVR having an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:392, 408, 424, 440, 456, 472, 488, 496, 504, 512, 520, 528, 544, 552, 560, 568, 576, 584, and 592, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity.

[0152] In another embodiment, an anti-Activin A antibody or antigen-binding fragment thereof comprises a LCVR having an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 400, 416, 432, 448, 464, 480, 536, and 600, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity.

[0153] In another embodiment, an anti-Activin A antibody or antigen-binding fragment thereof comprises a HCVR and a LCVR, wherein the HCVR/LCVR pair has amino acid sequences encoded by a nucleic acid sequence pair selected from the group consisting of SEQ ID NO: 392/400, 408/416, 424/432, 440/448, 456/464, 472/480, 488/480, 496/480, 504/480, 512/480, 520/480, 528/536, 544/536, 552/536, 560/536, 568/536, 576/536, 584/536, and 592/600.

[0154] In another embodiment, an anti-Activin A antibody or antigen-binding fragment thereof comprises a HCDR3 domain having an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:398, 414, 430, 446, 462, 478, 498, 502, 510, 518, 526, 534, 550, 558, 566, 574, 582, 590, and 598, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity, and a LCDR3 domain having an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:406, 422, 435, 454, 470, 486, 542, and 606, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity. In one embodiment, the HCDR3/LCDR3 domain set has amino acid sequences encoded by a nucleic acid sequence pair selected from the group consisting of SEQ ID NO:398/406, 414/422, 430/438, 446/454, 462/470, 478/486, 494/486, 502/486, 510/486, 518/486, 526/486, 534/542, 550/542, 558/542, 566/542, 574/542, 582/542, 590/542, and 598/606.

[0155] In another embodiment, an anti-Activin A antibody or antigen-binding fragment thereof comprises HCDR1 and HCDR2 domains, and LCDR1 and LCDR2 domains, wherein the HCDR1 domain has an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:394, 410, 426, 442, 458, 474, 490, 498, 506, 514, 522, 530, 546, 554, 562, 570, 578, 586, and 594, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity, the HCDR2 domain has an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:396, 412, 428, 444, 460, 476, 492, 500, 508, 516, 524, 532, 548, 556, 564, 572, 580, 588, and 596, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity, the LCDR1 domain has an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:402, 418, 434, 450, 466, 482, 538, and 602, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity, and the LCDR2 domain has an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:404, 420, 436, 452, 468, 484, 540, and 604, or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity.

[0156] In another embodiment, an anti-Activin A antibody or antigen-binding fragment thereof comprises heavy and light chain CDR (HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, LCDR3) domains having amino acid sequences encoded by a nucleic acid sequence set selected from the group consisting of SEQ ID NO: 394/396/398/402/404/406, 410/412/414/418/420/422, 426/428/430/434/436/438, 442/444/446/450/452/454, 458/460/462/466/468/470, 474/476/478/482/484/486, 490/492/494/482/484/486, 498/500/502/482/484/486, 506/508/510/482/484/486, 514/516/518/482/484/486, 522/524/526/482/484/486, 530/532/534/538/540/542, 546/548/550/538/540/542, 554/556/558/538/540/542, 562/564/566/538/540/542, 570/572/574/538/540/542, 578/580/582/538/540/542, 586/588/590/538/540/542, and 594/596/598/602/604/606.

[0157] In one embodiment, the anti-Activin A antibody or antigen-binding fragment thereof comprises the HCVR and LCVR (HCVR/LCVR) amino sequence pair of SEQ ID NO: 553/537 and the anti-GDF8 antibody or antigen-binding fragment thereof comprises a HCVR and LCVR (HCVR/LCVR) amino sequence pair of SEQ ID NO: 360/368.

[0158] In another embodiment, the anti-Activin A antibody or antigen-binding fragment thereof comprises HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 domains, respectively, having the amino acid sequences of: SEQ ID NOs: 555-557-559-539-541-543 (H4H10446P2), and the anti-GDF8 antibody or antigen-binding fragment thereof comprises HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 domains, respectively, having the amino acid sequences of: SEQ ID NOs: 362/364/366/370/372/374 (e.g., H4H1657N2).

[0159] In another embodiment, an anti-Activin A antibody or antigen-binding fragment thereof has a modified glycosylation pattern. In some applications, modification to remove undesirable glycosylation sites may be useful, or an antibody lacking a fucose moiety present on the oligosaccharide chain, for example, to increase antibody dependent cellular cytotoxicity (ADCC) function (Shield et al. (2002) JBC 277:26733). In other applications, modification of a galactosylation can be made in order to modify complement dependent cytotoxicity (CDC).

[0160] The fully-human anti-Activin A and/or anti-GDF8 antibodies described herein may comprise one or more amino acid substitutions, insertions and/or deletions in the framework and/or CDR regions of the heavy and light chain variable domains as compared to the corresponding germline sequences. Such mutations can be readily ascertained by comparing the amino acid sequences disclosed herein to germline sequences available from, for example, public antibody sequence databases. The compositions and methods of the invention use, in additional embodiments, antibodies and antigen-binding fragments thereof that are derived from any of the amino acid sequences disclosed herein, wherein one or more amino acids within one or more framework and/or CDR regions are back-mutated to the corresponding germline residue(s) or to a conservative amino acid substitution (natural or non-natural) of the corresponding germline residue(s) (such sequence changes are referred to herein as "germline back-mutations"). A person of ordinary skill in the art, starting with the heavy and light chain variable region sequences described herein, can easily produce numerous antibodies and antigen-binding fragments that comprise one or more individual germline back-mutations or combinations thereof. In certain embodiments, all of the framework and/or CDR residues within the V.sub.H and/or V.sub.L domains are mutated back to the germline sequence. In other embodiments, only certain residues are mutated back to the germline sequence, e.g., only the mutated residues found within the first 8 amino acids of FR1 or within the last 8 amino acids of FR4, or only the mutated residues found within CDR1, CDR2 or CDR3. Furthermore, the antibodies and antigen-binding fragments used in the compositions and methods of the invention may contain any combination of two or more germline back-mutations within the framework and/or CDR regions, i.e., wherein certain individual residues are mutated back to the germline sequence while certain other residues that differ from the germline sequence are maintained. Once obtained, antibodies and antigen-binding fragments that contain one or more germline back-mutations can be easily tested for one or more desired property such as, improved binding specificity, increased binding affinity, improved or enhanced antagonistic or agonistic biological properties (as the case may be), reduced immunogenicity, etc. Antibodies and antigen-binding fragments obtained in this general manner are encompassed within the invention.

[0161] The compositions and methods of the invention use, in additional embodiments, anti-GDF8 antibodies and/or anti-Activin A antibodies (or antigen-binding fragments thereof) comprising variants of any of the HCVR, LCVR, and/or CDR amino acid sequences described herein having one or more conservative substitutions. For example, anti-GDF8 antibodies and/or anti-Activin A antibodies used in the compositions and methods of the invention have, in some embodiments, HCVR, LCVR, and/or CDR amino acid sequences with, e.g., 10 or fewer, 8 or fewer, 6 or fewer, 4 or fewer, etc. conservative amino acid substitutions relative to any of the HCVR, LCVR, and/or CDR amino acid sequences described herein.

Bispecific Antibodies

[0162] Bispecific antibodies (bsAbs) combine specificities of two antibodies and simultaneously bind different antigens or epitopes. Two or more antigen-recognizing elements are engineered into a single antibody. In one embodiment of the methods of the invention, the composition comprises an antibody comprising a GDF8-specific binding domain and an Activin A-specific binding domain. The term (antigen)"-specific binding domain," as used herein, includes polypeptides comprising or consisting of: (i) an antigen-binding fragment of an antibody molecule, (ii) a peptide that specifically interacts with a particular antigen (e.g., a peptibody), and/or (iii) a ligand-binding portion of a receptor that specifically binds a particular antigen. For example, included are bispecific antibodies with one arm comprising a first heavy chain variable region/light chain variable region (HCVR/LCVR) pair that specifically binds GDF8 and another arm comprising a second HCVR/LCVR pair that specifically binds Activin A.

[0163] Bispecific antibodies can be prepared according to known methods, including chemical cross-linking, hybrid hybridomas/quadromas, knobs into holes, CrossMab, dual-variable-domain immunoglobulin, recombinant engineering (tandem single chain variable fragments/diabodies), and dock and lock. Other exemplary bispecific formats that can be used in the context of the present invention include, without limitation, e.g., IgG-scFv fusions, dual variable domain (DVD)-Ig, common light chain, CrossFab, (SEED)body, leucine zipper, Duobody, IgG1/IgG2, dual acting Fab (DAF)-IgG, and Mab2 bispecific formats (see, e.g., Klein et al., mAbs 4:6, 1-11 (2012), and references cited therein, for a review of the foregoing formats). Bispecific antibodies can also be constructed using peptide/nucleic acid conjugation, e.g., wherein unnatural amino acids with orthogonal chemical reactivity are used to generate site-specific antibody-oligonucleotide conjugates, which then self-assemble into multimeric complexes with defined composition, valency and geometry. (See, e.g., Kazane et al., J Am Chem Soc. 135(1):340-346 (2013)).

Specific Binding

[0164] The term "specifically binds" or the like, as used herein, means that an antigen-specific binding protein, or an antigen-specific binding domain, forms a complex with a particular antigen characterized by a dissociation constant (K.sub.D) of 500 pM or less, and does not bind other unrelated antigens under ordinary test conditions. "Unrelated antigens" are proteins, peptides or polypeptides that have less than 95% amino acid identity to one another. Methods for determining whether two molecules specifically bind one another are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. For example, an antigen-specific binding protein or an antigen-specific binding domain, as used in the context of the present invention, includes antibodies or antigen-binding fragments thereof that bind a particular antigen (e.g., GDF8, Activin A) or a portion thereof with a K.sub.D of less than about 500 pM, less than about 400 pM, less than about 300 pM, less than about 200 pM, less than about 100 pM, less than about 90 pM, less than about 80 pM, less than about 70 pM, less than about 60 pM, less than about 50 pM, less than about 40 pM, less than about 30 pM, less than about 20 pM, less than about 10 pM, less than about 5 pM, less than about 4 pM, less than about 2 pM, less than about 1 pM, less than about 0.5 pM, less than about 0.2 pM, less than about 0.1 pM, or less than about 0.05 pM, as measured in a surface plasmon resonance assay.

[0165] Antibody binding (of antigen) can be quantitated in terms of K.sub.D, a measurement of affinity. The lower the K.sub.D value, the higher the binding affinity of the antibody. The term "K.sub.D", as used herein, is intended to refer to the equilibrium dissociation constant of a particular antibody-antigen interaction. Surface plasmon resonance can be used to measure ligand binding, for example, antibody-antigen interaction.

[0166] The term "surface plasmon resonance", as used herein, refers to an optical phenomenon that allows for the analysis of real-time biospecific interactions by detection of alterations in protein concentrations within a biosensor matrix, for example using the BIACORE.TM. system (Pharmacia Biosensor AB, Uppsala, Sweden and Piscataway, N.J.).

[0167] The term "epitope" includes any determinant, preferably a polypeptide determinant, capable of specific binding to an immunoglobulin or T-cell receptor. In certain embodiments, epitope determinants include chemically active surface groupings of molecules such as amino acids, sugar side chains, phosphoryl groups, or sulfonyl groups, and, in certain embodiments, may have specific three-dimensional structural characteristics, and/or specific charge characteristics. An epitope is a region of an antigen that is bound by an antibody. In certain embodiments, an antibody is said to specifically bind an antigen when it preferentially recognizes its target antigen in a complex mixture of proteins and/or macromolecules. For example, an antibody is said to specifically bind an antigen when the K.sub.D is less than or equal to 10.sup.-8 M, less than or equal to 10.sup.-9 M, or less than or equal to 10.sup.-10 M.

Preparation of Human Antibodies

[0168] Methods for generating monoclonal antibodies, including fully human monoclonal antibodies are known in the art. Any such known methods can be used in the context of the invention to make human antibodies that specifically bind to GDF8 and/or to Activin A.

[0169] Using VELOCIMMUNE.TM. technology or any other known method for generating monoclonal antibodies, high affinity chimeric antibodies to GDF8 and/or Activin A are initially isolated having a human variable region and a mouse constant region. As in the experimental section below, the antibodies are characterized and selected for desirable characteristics, including affinity, selectivity, epitope, etc. The mouse constant regions are replaced with a desired human constant region to generate the fully human antibody of the invention, for example wild-type or modified IgG1 or IgG4. While the constant region selected may vary according to specific use, high affinity antigen-binding and target specificity characteristics reside in the variable region.

[0170] In general, the antibodies used in the methods of the instant invention possess very high affinities, typically possessing K.sub.D of from about 10.sup.-12 through about 10.sup.-9 M, when measured by binding to antigen either immobilized on solid phase or in solution phase.

Pharmaceutical Compositions and Methods of Administration

[0171] The present invention includes methods for altering body composition of a subject. As used herein, the phrase "altering body composition" refers to a change in one or more of lean mass, fat mass, and/or bone mass in a subject. In some embodiments, body composition in a subject may be altered by administering an effective amount of a GDF8 inhibitor and an Activin A inhibitor to the subject. Lean mass may be, for example, thigh muscle volume, appendicular lean body mass, or total lean mass, etc. In some aspects, the thigh muscle volume may refer to thigh muscle tissue volume excluding intramuscular adipose tissue and large vessels. In some aspects, the thigh muscle volume may refer to thigh muscle tissue volume including intramuscular adipose tissue and large vessels. In some aspects, the appendicular lean body mass may be calculated by, for example, aLBM equation. In some aspects, the appendicular lean mass may be calculated by the sum of lean mass of arms and legs. Fat mass may be, for example, total fat mass, android fat mass, sum of intramuscular and perimuscular adipose tissue (IMAT), subcutaneous adipose tissue volume, sum of fat mass of arms and legs, thigh intramuscular adipose tissue, etc. Bone mass may be, for example, total bone mineral density(BMD) mass, total bone mineral content (BMC) mass, etc. In some embodiments, alteration of body composition comprises an increase in muscle mass and/or a reduction of fat mass. In some embodiments, alteration of body composition comprises an increase in muscle mass and a reduction of fat mass simultaneously. In some embodiments, alteration of body composition comprises an increase in muscle mass and a reduction of fat mass simultaneously, without reduction in bone mass. In some embodiments, alteration of body composition comprises an increase in bone mineral content mass. In some embodiments, alteration of body composition comprises a decrease in total fat mass, android fat mass, and/or subcutaneous fat mass. In some embodiments, alteration of body composition comprises a decrease in total fat mass, android fat mass, and/or subcutaneous fat mass, without a reduction in thigh intramuscular adipose tissue volume.

[0172] The present invention includes methods for altering body composition, for example, inducing a reduction in fat mass in a subject and methods for treating a disease or disorder characterized by increased fat mass, comprising administering a first composition comprising an effective amount of a GDF8 inhibitor and a second composition comprising an effective amount of an Activin A inhibitor to the subject. The first and second compositions can be administered concurrently or sequentially to the subject. The first and second compositions can also be combined into a third composition prior to administration. Thus, in certain embodiments, a composition comprising both a GDF8 inhibitor and an Activin A inhibitor can be administered to a subject. The GDF8 inhibitor in such a composition can, for example, be an anti-GDF8 antibody. The Activin A inhibitor in such a composition can, for example, be an anti-Activin A antibody.

[0173] The pharmaceutical compositions of the invention are formulated with suitable carriers, excipients, and other agents that provide suitable transfer, delivery, tolerance, and the like. A multitude of appropriate formulations can be found in the formulary known to all pharmaceutical chemists: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid (cationic or anionic) containing vesicles (such as LIPOFECTIN.TM.), DNA conjugates, anhydrous absorption pastes, oil-in-water and water-in-oil emulsions, emulsions carbowax (polyethylene glycols of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax. Any of the foregoing mixtures may be appropriate in treatments and therapies in accordance with the invention, provided that the active ingredient in the formulation is not inactivated by the formulation and the formulation is physiologically compatible and tolerable with the route of administration. See also Powell et al. "Compendium of excipients for parenteral formulations" PDA (1998) J Pharm Sci Technol 52:238-311.

[0174] Various delivery systems are known and can be used to administer the pharmaceutical compositions of the present invention, e.g., encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing the mutant viruses, receptor mediated endocytosis (see, e.g., Wu et al., 1987, J. Biol. Chem. 262:4429-4432). Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions may be administered by any convenient route, for example by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.) and may be administered together with other biologically active agents.

[0175] A pharmaceutical composition of the present invention can be delivered subcutaneously or intravenously with a standard needle and syringe. In addition, with respect to subcutaneous delivery, a pen delivery device readily has applications in delivering a pharmaceutical composition of the present invention. Such a pen delivery device can be reusable or disposable. A reusable pen delivery device generally utilizes a replaceable cartridge that contains a pharmaceutical composition. Once all of the pharmaceutical composition within the cartridge has been administered and the cartridge is empty, the empty cartridge can readily be discarded and replaced with a new cartridge that contains the pharmaceutical composition. The pen delivery device can then be reused. In a disposable pen delivery device, there is no replaceable cartridge. Rather, the disposable pen delivery device comes prefilled with the pharmaceutical composition held in a reservoir within the device. Once the reservoir is emptied of the pharmaceutical composition, the entire device is discarded.

[0176] Numerous reusable pen and autoinjector delivery devices have applications in the subcutaneous delivery of a pharmaceutical composition of the present invention. Examples include, but are not limited to AUTOPEN.TM. (Owen Mumford, Inc., Woodstock, UK), DISETRONIC.TM. pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75/25.TM. pen, HUMALOG.TM. pen, HUMALIN 70/30.TM. pen (Eli Lilly and Co., Indianapolis, Ind.), NOVOPEN.TM. I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR.TM. (Novo Nordisk, Copenhagen, Denmark), BD.TM. pen (Becton Dickinson, Franklin Lakes, N.J.), OPTIPEN.TM., OPTIPEN PRO.TM., OPTIPEN STARLET.TM., and OPTICLIK.TM. (Sanofi-Aventis, Frankfurt, Germany), to name only a few. Examples of disposable pen delivery devices having applications in subcutaneous delivery of a pharmaceutical composition of the present invention include, but are not limited to the SOLOSTAR.TM. pen (Sanofi-Aventis), the FLEXPEN.TM. (Novo Nordisk), and the KWIKPEN.TM. (Eli Lilly), the SURECLICK.TM. Autoinjector (Amgen, Thousand Oaks, Calif.), the PENLET.TM. (Haselmeier, Stuttgart, Germany), the EPIPEN (Dey, L.P.), and the HUMIRA.TM. Pen (Abbott Labs, Abbott Park Ill.), to name only a few.

[0177] In certain situations, the pharmaceutical compositions of the present invention can be delivered in a controlled release system. In one embodiment, a pump may be used (see Langer, supra; Sefton, 1987, CRC Crit. Ref. Biomed. Eng. 14:201). In another embodiment, polymeric materials can be used; see, Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Pres., Boca Raton, Fla. In yet another embodiment, a controlled release system can be placed in proximity of the composition's target, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, 1984, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138). Other controlled release systems are discussed in the review by Langer, 1990, Science 249:1527-1533.

[0178] The injectable preparations may include dosage forms for intravenous, subcutaneous, intracutaneous and intramuscular injections, drip infusions, etc. These injectable preparations may be prepared by known methods. For example, the injectable preparations may be prepared, e.g., by dissolving, suspending or emulsifying the antibody or its salt described above in a sterile aqueous medium or an oily medium conventionally used for injections. As the aqueous medium for injections, there are, for example, physiological saline, an isotonic solution containing glucose and other auxiliary agents, etc., which may be used in combination with an appropriate solubilizing agent such as an alcohol (e.g., ethanol), a polyalcohol (e.g., propylene glycol, polyethylene glycol), a nonionic surfactant [e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)], etc. As the oily medium, there are employed, e.g., sesame oil, soybean oil, etc., which may be used in combination with a solubilizing agent such as benzyl benzoate, benzyl alcohol, etc. The injection thus prepared is preferably filled in an appropriate ampoule.

[0179] Advantageously, the pharmaceutical compositions for oral or parenteral use described above are prepared into dosage forms in a unit dose suited to fit a dose of the active ingredients. Such dosage forms in a unit dose include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc.

Dosage

[0180] The amount of active ingredient (e.g., anti-GDF8 antibody and/or anti-Activin A antibody) that can be administered to a subject is, generally, a therapeutically effective amount. The term "effective amount" is a concentration or amount of an active ingredient, for example, an antibody or antigen-binding fragment of an antibody, which results in achieving a particular stated purpose. The term "effective amount" is used interchangeably with the term "therapeutically effective amount" and signifies a concentration or amount of an active ingredient, for example, an antibody or antigen-binding fragment thereof, which is effective for achieving a stated therapeutic effect. The (therapeutically) effective amount may be determined empirically.

[0181] As used herein, the phrase "therapeutically effective amount" or "effective amount" means a dose of antigen-specific binding proteins and/or antigen-binding molecules (e.g., antibodies) that results in a detectable decrease in fat mass. The effective amount may also, in certain embodiments, result in an increase in one or more of the following parameters: body weight, muscle mass (e.g., tibialis anterior [TA] muscle mass, gastrocnemius [GA] muscle mass, quadriceps [Quad] muscle mass, appendicular lean body mass, etc.), muscle volume (e.g., thigh muscle volume), muscle strength/power, and/or muscle function, and glucose tolerance.

[0182] A "therapeutically effective amount" or "effective amount" of a GDF8 inhibitor (e.g., anti-GDF8 antibody) and/or an Activin A inhibitor (e.g., anti-Activin A antibody) includes, e.g., an amount of GDF8 inhibitor and/or Activin A inhibitor that, when administered to a subject, causes a decrease in total fat mass of at least about 2% to 8%, at least 2.5% to 6%, at least 3% to 4%, or at least about 2.0%, at least about 2.5%, at least about 3.0%, or at least about 3.5%, or more. For example, a "therapeutically effective amount" or "effective amount" of a GDF8 inhibitor (e.g., anti-GDF8 antibody) and/or an Activin A inhibitor (e.g., anti-Activin A antibody) includes, e.g., an amount of GDF8 inhibitor and/or Activin A inhibitor that, when administered to a subject, causes a decrease in total fat mass of at least about 3.5% or more.

[0183] In some embodiments, a "therapeutically effective amount" or "effective amount" of a GDF8 inhibitor (e.g., anti-GDF8 antibody) and/or an Activin A inhibitor (e.g., anti-Activin A antibody) includes, e.g., an amount of GDF8 inhibitor and/or Activin A inhibitor that, when administered to a subject, causes a decrease in android fat mass of at least about 2% to 8%, at least 2.5% to 6%, at least 3% to 4%, or at least about 2.0%, at least about 2.5%, at least about 3.0%, or at least about 3.5%, or more. For example, a "therapeutically effective amount" or "effective amount" of a GDF8 inhibitor (e.g., anti-GDF8 antibody) and/or an Activin A inhibitor (e.g., anti-Activin A antibody) includes, e.g., an amount of GDF8 inhibitor and/or Activin A inhibitor that, when administered to a subject, causes a decrease in android fat mass of at least about 3.5%.

[0184] In certain embodiments, the amount also results in an increase in TA or GA muscle mass of at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60% or more, compared to control treated subjects.

[0185] In some embodiments, a "therapeutically effective amount" or "effective amount" of a GDF8 inhibitor (e.g., anti-GDF8 antibody) and/or an Activin A inhibitor (e.g., anti-Activin A antibody) includes, e.g., an amount of GDF8 inhibitor and/or Activin A inhibitor that, when administered to a subject, causes an increase in thigh muscle volume of at least about 2% to 8%, 2.5% to 6%, 3% to 4%, or at least 2.0%, at least 2.5%, at least 3.0%, or at least 3.5%, or more

[0186] In some embodiments, a "therapeutically effective amount" or "effective amount" of a GDF8 inhibitor (e.g., anti-GDF8 antibody) and/or an Activin A inhibitor (e.g., anti-Activin A antibody) includes, e.g., an amount of GDF8 inhibitor and/or Activin A inhibitor that, when administered to a subject, causes an increase in total lean body mass of at least about 2% to 8%, 2.5% to 6%, 3% to 4%, or at least 2.0%, at least 2.5%, at least 3.0%, or at least 3.5%, or more

[0187] In some embodiments, a "therapeutically effective amount" or "effective amount" of a GDF8 inhibitor (e.g., anti-GDF8 antibody) and/or an Activin A inhibitor (e.g., anti-Activin A antibody) includes, e.g., an amount of GDF8 inhibitor and/or Activin A inhibitor that, when administered to a subject, causes an increase in appendicular lean body mass of at least about 2% to 8%, 2.5% to 6%, 3% to 4%, or at least 2.0%, at least 2.5%, at least 3.0%, or at least 3.5%, or more.

[0188] In certain embodiments, the amount also results in an increase in TA or GA muscle mass of at least 2%, 3%, 5%, 10%, 15%, 20%, 25% or more, compared to control treated subjects.

[0189] In certain embodiments, a (therapeutically) effective amount of an anti-GDF8 antibody, anti-Activin A antibody, or bispecific antibody that specifically binds GDF8 and Activin A can be from about 0.05 mg to about 600 mg; e.g., about 0.05 mg, about 0.1 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, about 500 mg, about 510 mg, about 520 mg, about 530 mg, about 540 mg, about 550 mg, about 560 mg, about 570 mg, about 580 mg, about 590 mg, or about 600 mg, of the respective antibody. The dose may vary depending upon the age and the size of a subject to be administered, target disease, conditions, route of administration, and the like. Depending on the severity of the condition, the frequency and the duration of the treatment can be adjusted.

[0190] The amount of antibody (e.g., anti-GDF8 antibody, anti-Activin A antibody, or bispecific antibody that specifically binds GDF8 and Activin A) contained within the individual doses may be expressed in terms of milligrams of antibody per kilogram of patient body weight (i.e., mg/kg). For example, the anti-GDF8 antibody, anti-Activin A antibody, and/or anti-GDF8/anti-Activin A bispecific antibody in the first, second, or third composition administered per the methods of the invention may be administered to a patient at a dose of about 0.0001 to about 50 mg/kg of patient body weight (e.g. 0.5 mg/kg, 1.0 mg/kg, 1.5 mg/kg, 2.0 mg/kg, 2.5 mg/kg, 3.0 mg/kg, 3.5 mg/kg, 4.0 mg/kg, 4.5 mg/kg, 5.0 mg/kg, 5.5 mg/kg, 6.0 mg/kg, 6.5 mg/kg, 7.0 mg/kg, 7.5 mg/kg, 8.0 mg/kg, 8.5 mg/kg, 9.0 mg/kg, 9.5 mg/kg, 10.0 mg/kg, 10.5 mg/kg, 11.0 mg/kg, 11.5 mg/kg, etc.).

[0191] The effective amount of a GDF8 inhibitor (e.g., an anti-GDF8 antibody) may, in certain embodiments, comprises a dosing regimen selected from the group consisting of at least 0.1 mg/kg to about 10 gm/kg, 1 mg/kg to about 1 gm/kg, and 10 mg/kg to 100 mg/kg. The effective amount of an Activin A inhibitor (e.g., an anti-Activin A antibody) may, in certain embodiments, comprise a dosing regimen selected from the group consisting of at least 0.1 mg/kg to about 10 gm/kg, 1 mg/kg to about 1 gm/kg, and 10 mg/kg to 100 mg/kg.

[0192] The effective amount of a GDF8 inhibitor (e.g., an anti-GDF8 antibody) may, in additional embodiments, comprise a dosing regimen selected from a group consisting of a single dose of about 0.01 to about 20 mg/kg body weight, about 0.1 to about 10 mg/kg body weight, and about 0.1 to about 5 mg/kg body weight. The effective amount of an Activin A inhibitor (e.g., an anti-Activin A antibody) may, in additional embodiments, comprise a dosing regimen selected from a group consisting of a single dose of about 0.01 to about 20 mg/kg body weight, about 0.1 to about 10 mg/kg body weight, and about 0.1 to about 5 mg/kg body weight. In one specific aspect, the effective amount of an anti-GDF8 antibody is from about 2 mg/kg-10 mg/kg, 4 mg/kg-8 mg/kg, or about 6 mg/kg body weight of subject, and the effective amount of the anti-Activin A antibody is from 0.5 mg/kg-15 mg/kg, 2 mg/kg-12 mg/kg, about 3 mg/kg, or about 10 mg/kg body weight of a subject.

[0193] The first, second, and third compositions administered per methods of the invention may, in certain embodiments, comprise equal amounts of GDF8 inhibitor (e.g., anti-GDF8 antibody) and/or Activin A inhibitor (e.g., anti-Activin A antibody). Alternatively, the amount of GDF8 inhibitor (e.g., anti-GDF8 antibody) in the composition may be less than or greater than the amount of Activin A inhibitor (e.g., an anti-Activin A antibody). The effective amount of a GDF8 inhibitor (e.g., anti-GDF8 antibody) may be lower, when in combination with an Activin A inhibitor (e.g., an anti-Activin A antibody), than in a separate composition. The effective amount of an Activin A inhibitor (e.g., an anti-Activin A antibody) may be lower, when in combination with a GDF8 inhibitor (e.g., anti-GDF8 antibody), than in a separate composition. A person of ordinary skill in the art, using routine experimentation, will be able to determine the appropriate amounts of the individual components in the compositions necessary to produce a desired therapeutic effect.

Aspects of the Disclosure

[0194] The disclosure provides compositions, kits, and methods of using GDF8 inhibitors and Activin A inhibitors to reduce fat mass (induce a reduction in fat mass) in a subject. The disclosure also provides compositions, kits, and methods of using GDF8 inhibitors and Activin A inhibitors to treat diseases, disorders, and/or conditions associated with or characterized by increased fat mass in a subject. In preferred embodiments, the GDF8 inhibitor is an antibody or antigen-binding fragment thereof that specifically binds GDF8.

Therapeutic Methods

[0195] The present invention includes methods for altering body composition, for example, methods for inducing a reduction in fat mass in a subject, methods for increasing muscle mass in a subject, and methods for treating a disease or disorder characterized by increased fat mass, by specifically binding GDF8 and/or Activin A. For example, the present invention includes methods for inducing a reduction in fat mass in a subject, inducing an increase in muscle mass in a subject, and methods for treating a disease or disorder characterized by increased fat mass in a subject, by administering to the subject i) a composition comprising an anti-GDF8 antibody and a composition comprising an anti-Activin A antibody or ii) a composition comprising both an anti-GDF8 antibody and an anti-Activin A antibody or iii) a composition comprising a bispecific antibody comprising a first variable domain comprising a HCVR/LCVR pair that specifically binds GDF8 and a second variable domain comprising a HCVR/LCVR pair that specifically binds Activin A. Any of the GDF8 inhibitors and/or Activin A inhibitors disclosed or referred to herein can be used in the context of these aspects of the invention.

[0196] In methods comprising administering a GDF8 inhibitor and an Activin A inhibitor to a subject, the GDF8 inhibitor (for example, an anti-GDF8 antibody) and the Activin A inhibitor (for example, an anti-Activin A antibody) may be administered to the subject at the same or substantially the same time, e.g., in a single therapeutic dosage (third composition) or in two separate dosages (first and second compositions), which are administered simultaneously or within less than about 5 minutes of one another. Alternatively, the GDF8 inhibitor and the Activin A inhibitor (first and second compositions) may be administered to the subject sequentially, e.g., in separate therapeutic dosages separated in time from one another by more than about 5 minutes.

[0197] The reduction of fat mass in the subject of the methods according to the invention can be a reduction in total fat mass as measured by DXA (Dual-energy X-ray absorptiometry).

[0198] In another embodiment, the reduction of fat mass in the subject of the methods according to the invention is a reduction in android fat mass (i.e., visceral fat associated with the upper/central body) as measured by DXA (Dual-energy X-ray absorptiometry). In android obesity, the subject stores fat around his or her abdominal region. Android obesity can also be manifested in other areas of the upper trunk like the upper chest (front or back) nape area of the neck, and even the shoulders. Subjects who are android obese are at greater risk for obesity-related diseases/disorders like heart disease, and metabolic syndrome. The likelihood of developing gout, arterial-related diseases (due to high blood pressure) and many kinds of cancers are also linked to the central type of fat distribution in subjects who exhibit android obesity.

[0199] Body fat assessments are varied in precision and accuracy. Common anthropometric measures include: weight, waist circumference, and skinfold measurements using skin calipers. More complex methods include: bioelectrical impedance analysis (BIA), the BOD POD, and dual-energy X-ray absorptiometry (DEXA or DXA). DXA is especially accurate and valid, because it considers bone mineral content when estimating body fat and muscle. DEXA scans can evaluate different areas of fat distribution to determine the android/gynoid fat ratio, which is distinct from body mass index. DXA can measure total fat mass, total body muscle mass, visceral fat (fat around the organs) levels, intramuscular fat (fat between the muscles), total bone mineral density, and can even provide regional breakdowns. Finally, DXA can accurately assess the distribution of body fat associated with increased insulin resistance.

Avoidance of Side Effects

[0200] The present invention includes methods for altering body composition, for example, inducing a reduction in fat mass in a subject, and methods for treating a disease or disorder characterized by increased fat mass, comprising administering a GDF8 inhibitor and an Activin A inhibitor to the subject, without causing adverse side effects associated with the administration of molecules which bind multiple (e.g., 3 or more) ActRIIB ligands, for example, as set forth in U.S. Pat. No. 8,871,209. For example, the clinical molecule referred to as ACE-031 (Acceleron Pharma, Inc., Cambridge, Mass.) is a multimer consisting of the extracellular portion of ActRIIB fused to an IgG Fc domain (this molecule is also referred to herein as "ActRIIB-Fc"). ActRIIB-Fc binds GDF8 as well as other ActRIIB ligands such as, e.g., Activin A, Activin B, GDF11, BMP9, BMP10, and TGF.beta., and is known to cause various adverse side effects when administered to human patients. For example, administration of ACE-031 to postmenopausal women in a Phase Ib ascending dose study was shown to cause undesired increases in hemoglobin and decreases in FSH levels. In addition, a Phase II study of ACE-031 in pediatric patients with muscular dystrophy was discontinued due to adverse effects including nose and gum bleeding. Dilated blood vessels are also observed in patients treated with ActRIIB-Fc. Effect of ACE-031 in boys with Duchenne muscular dystrophy (DMD) demonstrated trends for increased lean body mass and reduced fat mass but non-muscle-related adverse events contributed to a decision to discontinue the study. (Campbell, et al. 2017 Muscle Nerve 55: 458-464). Specifically inhibiting GDF8 and Activin A (e.g., by administering an anti-GDF8 antibody and an anti-Activin A antibody), while not inhibiting other ActRIIB ligands such as Activin B, GDF11, BMP9, BMP10, and TGF.beta., results in an increase in a reduction in fat mass, without causing the adverse side effects associated with non-specific Activin-binding agents such as ActRIIB-Fc.

Administration Regimens

[0201] According to certain embodiments of the present invention, multiple doses of the compositions of the present invention (e.g., compositions comprising a GDF8 inhibitor and/or an Activin A inhibitor, for example, an anti-GDF8 antibody and/or an anti-Activin A antibody, or a bispecific antibody against GDF8 and Activin A), may be administered to a subject over a defined time course. The methods according to this aspect of the invention comprise sequentially administering to a subject multiple doses of the composition(s) of the present invention. As used herein, "sequentially administering" means that each dose of the compositions of the present invention are administered to the subject at a different point in time, e.g., on different days separated by a predetermined interval (e.g., hours, days, weeks or months). The present invention includes methods that comprise sequentially administering to the patient an initial dose of a first and/or a second composition; or a third composition; followed by one or more secondary doses of the first and/or second composition; or the third composition; and optionally followed by one or more tertiary doses of the first and/or second composition; or the third composition.

[0202] The terms "initial dose," "secondary doses," and "tertiary doses," refer to the temporal sequence of administration of the compositions of the present invention. Thus, the "initial dose" is the dose that is administered at the beginning of the treatment regimen (also referred to as the "baseline dose"); the "secondary doses" are the doses that are administered after the initial dose; and the "tertiary doses" are the doses that are administered after the secondary doses. The initial, secondary, and tertiary doses may all contain the same amount of active ingredient(s), e.g., anti-GDF8 antibody and/or anti-Activin A antibody, but will generally differ from one another in terms of frequency of administration. In certain embodiments, however, the amount of active ingredient(s) contained in the initial, secondary and/or tertiary doses will vary from one another (e.g., adjusted up or down as appropriate) during the course of treatment.

[0203] In one exemplary embodiment of the present invention, each secondary and/or tertiary dose is administered 1 to 30 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more) days after the immediately preceding dose. The phrase "the immediately preceding dose," as used herein, means, in a sequence of multiple administrations, the dose(s) of the compositions of the present invention that are administered to a subject prior to the administration of the very next dose in the sequence, with no intervening doses.

[0204] The methods according to this aspect of the invention may comprise administering to a patient any number of secondary and/or tertiary doses of the compositions of the present invention. For example, in certain embodiments, only a single secondary dose is administered to the patient. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8, or more) secondary doses are administered to the patient. Likewise, in certain embodiments, only a single tertiary dose is administered to the patient. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8, or more) tertiary doses are administered to the patient.

[0205] In embodiments involving multiple secondary doses, each secondary dose may be administered at the same frequency as the other secondary doses. For example, each secondary dose may be administered to the patient 1 to 29 days after the immediately preceding dose. Similarly, in embodiments involving multiple tertiary doses, each tertiary dose may be administered at the same frequency as the other tertiary doses. For example, each tertiary dose may be administered to the patient 1 to 60 days after the immediately preceding dose. Alternatively, the frequency at which the secondary and/or tertiary doses are administered to a patient can vary over the course of the treatment regimen. The frequency of administration may also be adjusted during the course of treatment by a physician, depending on the needs of the individual patient following clinical examination.

[0206] In one embodiment, a subject may be subjected to preliminary DXA, then receive a composition comprising an anti-GDF8 antibody and an anti-Activin A antibody (or a composition comprising an anti-GDF8 antibody and a composition comprising an anti-Activin A antibody), then be subjected to follow-up DXA. If the fat mass is not measurably reduced in the follow-up DXA (in comparison with the preliminary DXA), the subject may receive the composition(s) again. Subsequent dosage amount and frequency of administration can, in an additional embodiment, be varied based on the results of the follow-up DXA.

Combination Therapies

[0207] The methods of the present invention, according to certain embodiments, comprise administering to the subject one or more additional therapeutic agents that may be advantageously combined with the composition comprising a GDF8 inhibitor and/or an Activin A inhibitor. As used herein, the expression "in combination with" means that the additional therapeutic agent(s) is/are administered before, after, or concurrently with a pharmaceutical composition comprising a GDF8 inhibitor and/or an Activin A inhibitor. The term "in combination with" also includes sequential or concomitant administration of a GDF8 inhibitor, an Activin A inhibitor, or both and a second therapeutic agent. The term "therapeutic agent" is also meant to include a specific therapy.

[0208] The additional therapeutic agent may be, e.g., another GDF8 antagonist/inhibitor, another Activin A antagonist/inhibitor, growth factor inhibitors, immunosuppressants, metabolic inhibitors, enzyme inhibitors, and cytotoxic/cytostatic agents, an IL-1 antagonist (including, e.g., an IL-1 antagonist as set forth in U.S. Pat. No. 6,927,044), an IL-6 antagonist, an IL-6R antagonist (including, e.g., an anti-IL-6R antibody as set forth in U.S. Pat. No. 7,582,298), an IL-13 antagonist, a tumor necrosis factor (TNF) antagonist, an IL-8 antagonist, an IL-9 antagonist, an IL-17 antagonist, an IL-5 antagonist, an IgE antagonist, a CD48 antagonist, an IL-31 antagonist (including, e.g., as set forth in U.S. Pat. No. 7,531,637), a thymic stromal lymphopoietin (TSLP) antagonist (including, e.g., as set forth in US 2011/027468), interferon-gamma (IFN.gamma.) antibiotics, topical corticosteroids, tacrolimus, pimecrolimus, cyclosporine, azathioprine, methotrexate, cromolyn sodium, proteinase inhibitors, systemic corticosteroids, systemic immunotherapy, anti-histamines, chemotherapy, light therapy, or combinations thereof.

[0209] In further embodiments, the invention features a composition, the additional therapeutic agent is selected from the group consisting of (1) 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors, such as cerivastatin, atorvastatin, simvastatin, pitavastatin, rosuvastatin, fluvastatin, lovastatin, pravastatin, and the like; (2) inhibitors of cholesterol uptake and/or bile acid re-absorption; (3) niacin, which increases lipoprotein catabolism; (4) fibrates or amphipathic carboxylic acids, which reduce low-density lipoprotein (LDL) level, improve high-density lipoprotein (HDL) and TG levels, and reduce the number of non-fatal heart attacks; and (5) activators of the LXR transcription factor that plays a role in cholesterol elimination such as 22-hydroxycholesterol, or fixed combinations such as ezetimibe plus simvastatin; a statin with a bile resin (e.g., cholestyramine, colestipol, colesevelam), a fixed combination of niacin plus a statin (e.g., niacin with lovastatin); or with other lipid lowering agents such as omega-3-fatty acid ethyl esters (for example, omacor).

[0210] In still further embodiments, the second therapeutic agent is selected from one or more other inhibitors/antagonists of glucagon or an inhibitor/antagonist of the glucagon receptor, as well as inhibitors of other molecules, such as inhibitors of ANGPTL8 (for example, an anti-ANGPTL8 antibody), as well as inhibitors of other molecules, such as ANGPTL3 (for example, an anti-ANGPTL3 antibody), ANGPTL4, ANGPTL5, ANGPTL6, apolipoprotein C-III (also referred to as APOC3; see for example, inhibitors of APOC3 described in U.S. Pat. Nos. 8,530,439, 7,750,141, 7,598,227 and volanesorsen, also referred to as ISIS-APOCIIIRx) and proprotein convertase subtilisin/kexin type 9 (PCSK9), which are involved in lipid metabolism, in particular, cholesterol and/or triglyceride homeostasis. Inhibitors of these molecules include small molecules, antisense molecules and antibodies that specifically bind to these molecules and block their activity.

[0211] The additional therapeutic agent may, in further embodiments, be selected from the group consisting of analgesics, anti-inflammatory agents, including non-steroidal anti-inflammatory drugs (NSAIDS), such as Cox-2 inhibitors, and the like, so as to ameliorate and/or reduce the symptoms accompanying the underlying condition, if needed.

[0212] The additional therapeutic agent(s) may be administered prior to, concurrent with, or after the administration of the first and/or second; or third composition(s) described herein. For purposes of the present disclosure, such administration regimens may be considered the administration of an anti-GDF8 antibody and/or an anti-Activin A antibody "in combination with" a second therapeutically active component.

Examples

[0213] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the methods and compositions of the invention, and are not intended to limit the scope of what the inventors regard as their invention. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.), but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric.

Example 1. Anti-GDF8 Alone Increased Total Lean Mass Up to 3% in Clinical Studies: Sarcopenia Phase 2 Data

[0214] A randomized, double-blind, placebo-controlled interventional study phase 2 clinical trial was performed in patients 70 years and older including men and postmenopausal women having sarcopenia. Patients were treated 12 weeks with subcutaneous anti-GDF8 antibody REGN1033 (H4H1657N2) alone, at either 100 mg anti-GDF8 antibody, Q4W s.c. (n=62); 300 mg anti-GDF8 antibody Q4W s.c.(n=64), or 300 mg anti-GDF8 antibody Q2W s.c. (n=59), or placebo (n=65). As shown in FIG. 1 significant increase in total lean body mass compared to placebo was exhibited in patients after 12 weeks when using REGN1033 alone at each of the three doses, as shown in FIG. 1 and Table 4.

TABLE-US-00003 TABLE 4 Total Lean Mass % Change to Week 12 using anti-GDF8 REGN1033 alone Anti-GDF8 Placebo 100 Q4W 300 Q4W 300 Q2W N 65 62 64 59 Baseline 43.6 kg 42.9 kg 42.7 42.8 Mean Difference vs. -- 1.66% 1.78% 2.29% placebo P value -- 0.0077 0.0043 0.0004

[0215] The effects on strength and function were varied. Anti-GDF8 was generally safe and well-tolerated (reactions, if any, were mild). Table 4 shows patients receiving REGN1033 in either 100 mg or 300 mg doses exhibited significantly increased Total lean mass as % change from placebo to week 12 data. Anti-GDF8 antibody REGN1033 alone increased total lean body mass up to 3% in the study. The 300 mg regimens also resulted in decreases in total and android fat mass.

Example 2. Combination of Anti-GDF8 and Anti-Activin a First In-Human Single Ascending Dose Study

[0216] A randomized, double-blind placebo-controlled, ascending dose study was initiated to assess the tolerability and effects on body composition of a combination of an intravenous anti-GDF8 antibody and an anti-Activin A antibody vs. the individual components.

[0217] The primary objective of the study was to assess the safety and tolerability of an anti-Activin A antibody (e.g., H4H10446P2=REGN2477) alone and combined with an anti-GDF8 antibody (e.g., H4H1657N2=REGN1033) in healthy postmenopausal women aged 45 to 70 years of age.

[0218] Secondary objectives of the study included: an assessment of the effect of REGN2477 alone, REGN1033 alone, and REGN2477+REGN1033 in combination on thigh muscle volume as measured by Magnetic Resonance Imaging (MRI), and an assessment of the effects of REGN2477 alone, REGN1033 alone and REGN2477+REGN1033 in combination on total and regional body composition as measured by dual-energy X-ray absorptiometry (DXA)

Study Design

[0219] This study was a randomized, double-blind, placebo-controlled, ascending dose study to assess the safety, tolerability, and pharmacodynamics of intravenous REGN2477 (anti-Activin A) alone and in combination with REGN1033 (anti-GDF8) in healthy postmenopausal women.

[0220] A total of 48 subjects were randomized to 1 of the following 4 sequential ascending REGN2477 IV dose panels where 8 subjects were randomized in a 6:2 ratio into each of the first 3 Panels (Panels A, B, and C) and 24 subjects were randomized in a 1:1:1:1 ratio (6 subjects each) into Panel D.

[0221] Panel A: 4 subjects REGN1033 (6 mg/kg IV)+REGN2477 low dose (1 mg/kg IV) or 2 subjects placebo

[0222] Panel B: 4 subjects REGN1033 (6 mg/kg IV)+REGN2477 medium dose (3 mg/kg IV) or 2 subjects placebo

[0223] Panel C: 4 subjects REGN1033 (6 mg/kg IV)+REGN2477 high dose (10 mg/kg IV) or 2 subjects placebo

[0224] Panel D: REGN1033(6 mg/kg IV+REGN2477 high dose (10 mg/kg IV) placebo, REGN2477 (10 mg/kg IV), or REGN1033 (6 mg/kg IV)

[0225] Subjects received a single intravenous dose of one or both of anti-GDF8 antibody REGN1033 and/or anti-Activin A antibody REGN2477. In the primary analyses, the placebo and high dose combination groups were pooled across panels, yielding 12 subjects on placebo and 12 on the high dose combination, as shown in FIG. 2A. The dosing schedule shown in FIG. 2A was used in studies shown in each of the subsequent FIGS. 2B to 17.

[0226] Subjects participated in a screening period of up to 28 days, followed by a baseline and treatment visit on day 1, and a follow-up period of 113 days.

Efficacy and Safety Analysis

[0227] The full analysis set (FAS) includes all randomized subjects; it is based on the treatment allocated (as randomized). Efficacy endpoints were analyzed using the FAS. The safety analysis set (SAF) includes all randomized subjects who received any study drug; it is based on the treatment received (as treated). Treatment compliance/administration and all clinical safety variables were analyzed using the SAF.

[0228] The efficacy variables included: Thigh muscle tissue volume, excluding and including intramuscular adipose tissue and large vessels as measured by Magnetic Resonance Imaging (MRI);Total lean mass as measured by dual X-ray absorptiometry (DXA); Appendicular lean body mass (calculated by aLBM equation) as measured by DXA; and Total fat mass as measured by DXA.

[0229] The demographics and baseline characteristics of the subjects were balanced across the treatment groups, as per the below Table 5:

TABLE-US-00004 TABLE 5 Baseline Characteristics were balanced across treatment groups Anti-GDF8 + Anti-GDF8 + Anti-GDF8 + Anti-Activin A anti-activin A anti-activin A anti-activin A Placebo Anti-GDF8 (10 mg/kg) IV (1 mg/kg) IV (3 mg/kg) IV (10 mg/kg) IV (N = 12) (N = 6) (N = 6) (N = 6) (N = 6) (N = 12) Age (Years) Median 54 56 61 60 60 55.5 Height (cm) Median 162.5 164.2 162.5 166.5 165 163.5 Weight (kg) Median 68.35 70.5 69.6 61.35 67.05 70.4 BMI (kg/m.sup.2) Median 26.2 26.55 26.65 22.9 25.55 26.6

Statistical Methods

[0230] The percent change and change of efficacy variables from baseline to either week 4 or 8 in the full analysis set (FAS) were analyzed using analysis of covariance (ANCOVA) model with treatment group as fixed effect, and the baseline value as continuous covariate. Least-squares means at week 4 and week 8 for each treatment group with the corresponding standard error, confidence interval, and the p value for treatment comparisons were provided from this model. Placebo subjects were pooled across panels. Missing efficacy data was not imputed. No adjustment for multiple testing is applied in this study.

Results

[0231] A total of 48 subjects were randomized, administered study drug and completed the study. One subject in the REGN2477+REGN1033 high dose group had an interrupted infusion of study drug due to an adverse event of `Infusion site swelling`.

Efficacy

[0232] The blockade of both Activin A and GDF8 in combination increased thigh muscle volume and decreased fat mass at week 8, as shown in FIGS. 2B and 2C. The greatest effect was seen at the highest dose combination.

[0233] FIG. 2B shows a bar graph showing % change in thigh muscle volume by MRI at 8 weeks after a single dose in postmenopausal women. Numbers show changes from placebo. * indicates nominal p<0.05 vs. placebo; **** indicates nominal p<0.0001 vs. placebo. The single dose amounts as mg/kg of either anti-Activin A antibody and/or anti-GDF8 antibody are shown below the bar graph. Treatment with REGN2477+REGN1033 in medium and high dose combinations resulted in significantly increased thigh muscle volume (p<0.05; p<0.0001, respectively) in a dose-related manner compared with placebo. Subjects in high dose panel exhibited up to 7.73% as % change from baseline versus 0.88% with placebo at week 8.

[0234] FIG. 2C shows a bar graph showing % change in total fat mass by DXA at 8 weeks after a single dose in postmenopausal women. Numbers show changes from placebo. * indicates nominal p<0.05 vs. placebo; **** indicates nominal p<0.0001 vs. placebo. The single dose amounts as mg/kg of either anti-Activin A antibody and/or anti-GDF8 antibody are shown below the bar graph. Treatment with high dose REGN2477+REGN1033 combination resulted in significantly decreased total fat mass (p<0.05) compared with placebo.

[0235] As shown in FIG. 3, anti-Activin A antibody REGN2477 combined with anti-GDF8 antibody REGN1033 led to dose-dependent increases in thigh muscle volume. The thigh muscle volume results, as measured via MRI, excluding intermuscular adipose tissue and large vessels, are likewise summarized in Table 6, below. Treatment with mid- and high dose REGN2477+REGN1033 combination resulted in significantly increased thigh muscle volume (p<0.05) compared with placebo, as shown in Table 6.

TABLE-US-00005 TABLE 6 Thigh Muscle Volume measured by MRI* anti-GDF8 + anti-GDF8 + anti-GDF8 + anti- anti-activin A anti-activin A anti-activin A anti-activin A Placebo GDF8 High Dose Low Dose Mid Dose High Dose Week 4, N 12 6 6 6 6 12 % change mean 0.4 3.3 0.3 4.9 4.6 5.9 Week 8, mean 0.9 4.6 2.9 3.4 6.2 7.8 % change .DELTA. LSM vs 3.73 (1.819) 1.97 (1.821) 2.63 (1.824) 5.31 (1.817) 6.85 (1.484) placebo (SE) Nominal 0.0467 0.2846 0.1569 0.0056 <0.0001 p-value *excluding intermuscular adipose tissue and large vessels

[0236] As shown in Table 4, treatment with REGN2477+REGN1033 in medium and high dose combinations resulted in significantly increased thigh muscle volume in a dose-related manner compared with placebo.

[0237] FIG. 4 shows that increases in thigh muscle volume were consistently observed in individual subjects following treatment with anti-Activin A+anti-GDF8 in combination. Within each treatment group, different lines indicate different individuals.

[0238] The pattern of effects on appendicular lean mass (the sum of lean mass of arms and legs) with anti-Activin A antibody REGN2477+anti-GDF8 antibody REGN1033 in combination was similar to that seen on thigh muscle volume, as shown in FIG. 5. The appendicular lean mass results, as measured via DXA, are likewise summarized in Table 7. The mid and high dosing combinations significantly increased appendicular lean mass compared to placebo, as shown in Table 7.

TABLE-US-00006 TABLE 7 Appendicular Lean Mass by DXA Anti-GDF8 + Anti-GDF8 + Anti-GDF8 + Anti-activin A anti-activin A anti-activin A anti-activin A Placebo Anti-GDF8 High Dose Low Dose Mid Dose High Dose N 12 6 6 6 6 12 Baseline, kg Mean 17.44 18.10 18.02 17.92 18.67 17.32 Week 4, Mean -0.2 1.2 2.1 4.2 4.6 3.8 % Change Week 8, Mean 0.8 2.6 2.3 3.7 6.9 5.8 % Change .DELTA.LSM vs 1.90 (1.266) 1.61 (1.265) 2.96 (1.264) 6.39 (1.275) 4.97 (1.031) Placebo (SE) Nominal 0.1418 0.2109 0.0242 <0.0001 <0.0001 p-value

[0239] As shown in FIG. 6, blockade of both Activin A and GDF8 led to reductions in total fat mass, as assessed by DXA. The high dose combination of anti-GDF8 antibody REGN1033 and anti-Activin A antibody REGN2477 significantly reduced total fat mass by DXA compared to placebo at week 8 (*p<0.05). The total fat mass results, as measured via DXA, are likewise summarized in Table 8, below.

TABLE-US-00007 TABLE 8 Total Fat Mass by DXA anti-GDF8 + anti-GDF8 + anti-GDF8 + anti-GDF8 anti-activin A anti-activin A anti-activin A anti-activin A Dose High High Low Mid High Week 8, N 6 6 6 6 12 % Change .DELTA.LSM vs -2.11 (1.96) -0.69 (1.93) -0.2 (1.93) -2.08 (1.93) -3.92 (1.58)* Placebo (SE) *p < 0.05

[0240] Blockade of Activin A and GDF8 was also found to be associated with decreases in android fat mass, as assessed by DXA, as shown in FIG. 7. The high dose combination of anti-GDF8 antibody REGN1033 and anti-Activin A antibody REGN2477 significantly reduced android fat mass by DXA compared to placebo at week 8 (*p<0.05).

[0241] Further efficacy results are shown in FIGS. 8 to 17 and summarized in the tables, below. REGN2477+REGN1033 high, medium and low doses are shown in FIG. 2A.

[0242] Compared with placebo, REGN2477+REGN1033 medium (p<0.05) and high dose groups (p<0.001) exhibited significantly increased thigh muscle volume, excluding Intramuscular adipose tissue and large vessels (FIG. 8). Thigh muscle volume increased in the REGN2477+REGN1033 high dose group by 7.73% as compared with 0.88% in the placebo group (nominal p<0.001) at 8 weeks. Compared with placebo, REGN2477+REGN1033 at the medium dose and REGN1033 alone also significantly increased thigh muscle volume. Increases in thigh muscle volume were consistently observed in individual subjects treated with the combination in a dose responsive manner. (data not shown).

[0243] Compared with placebo, REGN2477+REGN1033 high dose group exhibited significantly increased total lean mass by DXA (p<0.05) (FIG. 9).

[0244] Appendicular lean body mass (calculated via aLBM equation), was significantly increased in each of the combination REGN2477+REGN1033 treatment groups compared with placebo (low dose p<0.05, medium and high dose groups p<0.001)(FIG. 10). Appendicular lean body mass increased in the REGN2477+REGN1033 medium dose group by 7.15% from baseline as compared with 0.76% in the placebo group at 8 weeks. Similarly, appendicular lean body mass increased in the REGN2477+REGN1033 high dose group by 5.7% from baseline as compared with 0.76% in the placebo group.

[0245] Total fat mass was significantly decreased in the high dose REGN2477+REGN1033 treatment group; total fat mass was decreased: 3.92% (high dose group) compared with placebo at 0.5% (nominal p<0.05)(FIG. 11).

[0246] Compared to placebo, thigh muscle volume including intramuscular adipose tissue and large vessels was significantly increased in each of the medium and high dose REGN2477+REGN1033 groups and REGN1033 group at 4 weeks, and at 8 weeks (FIG. 12). Low dose REGN2477+REGN1033 also exhibited significant increase in thigh muscle volume including intramuscular adipose tissue and large vessels compared to placebo at 4 weeks.

[0247] Appendicular lean mass (sum of arms and legs) was significantly increased in each of combination REGN2477+REGN1033 treatment groups (p<0.05) at 4 and 8 weeks compared to placebo (FIG. 13).

[0248] Android fat mass in the high dose REGN2477+REGN1033 treatment group was also significantly reduced (FIG. 14). Android fat mass was reduced 6.6% in the high dose REGN2477+REGN1033 group as compared to no reduction in the placebo.

[0249] Thigh intramuscular adipose tissue volume (cm3) was significantly increased in the high dose REGN2477+REGN1033 group (p<0.05) as compared to placebo at 8 weeks (FIG. 15).

[0250] Reductions in adipose tissue in the high dose group were observed in the sum of Intramuscular and Perimuscular Adipose Tissue (IMAT)(FIG. 16) and in the low and medium dose group of subcutaneous adipose tissue. (FIG. 17). In contrast, thigh intramuscular adipose tissue was increased in the high dose group to 8% as compared with placebo with a reduction of 4%. (FIG. 17).

[0251] REGN2477+REGN1033, in the high dose group, significantly increased all of the major measures of muscle volume and lean mass compared with placebo, at both Week 4 and Week 8; the effects at Week 4 were generally less pronounced than at Week 8 (Table 9). A summary of Percent Change in Key Body Composition Measures at Week 8 (Full Analysis Set, LS Means and SE presented) is shown in Table 9, below.

TABLE-US-00008 TABLE 9 Summary of Change in Body Composition at Week 8 Low Medium High R2477 R2477 R2477 R1033 R2477 (1 mg/kg) (3 mg/kg) (10 mg/kg) Placebo (6 mg/kg) (10 mg/kg) +R1033 +R1033 +R1033 Endpoint (n = 12) (n = 6) (n = 6) (n = 6) (n = 6) (n = 12) Thigh % Change 0.88 (105) 4.61 (1.49) 2.85 (1.49) 3.51 (1.49) 6.19 (1.48) 7.73 (1.05) Muscle from Volume baseline Difference 3.73 (1.82)* 1.97 (1.82) 2.63 (1.82) 5.31 (1.82)* 6.85 (1.48)** from placebo Total Lean % Change 1.31 (0.64) 2.18 (0.89) 1.94 (0.90) 3.16 (0.89) 4.67 (0.89) 4.31 (0.63) Mass by from DXA baseline Difference 0.88 (1.10) 0.63 (1.11) 1.85 (1.10) 3.37 (1.10)* 3.00 (0.89)* from placebo Appendicular % Change 0.76 (0.73) 2.65 (1.03) 2.37 (1.03) 3.72 (1.03) 7.15 (1.04) 5.72 (0.73) Lean Body from Mass baseline Difference 1.90 (1.27) 1.61 (1.26) 2.96 (1.26)* 6.39 (1.27)** 4.97 (1.03)** from placebo Total Fat % Change -0.65 (1.12) -2.76 (1.60) -1.34 (1.58) -0.85 (1.58) -2.73 (1.58) -4.57 (1.12) Mass by from DXA baseline Difference -2.11 (1.96) -0.69 (1.93) -0.2 (1.93) -2.08 (1.93) -3.92 (1.58)* from placebo *p < 0.05; **p < 0.001

[0252] In Table 9, above, changes from baseline and differences from placebo are Least-Squares (LS) means based on the ANCOVA model with baseline as a covariate and treatment as a fixed factor. Standard errors (SE) and p-values also taken from the ANCOVA. Nominal p-values are reported.

[0253] Bone mineral density (BMD) mass and bone mineral content (BMC) mass were measured by DXA, as shown in Table 10. At the high dose, R2477+R1033 increased Bone Mineral Content as measured by DXA, while total bone mineral density did not change (Table 10).

[0254] Sum of fat mass of arms and legs were measured as shown in Table 10. At the high dose, R2477+R1033 decreased sum of fat mass of arms and legs (p<0.05).

[0255] The data from the primary endpoint analysis is shown in Table 10, below.

TABLE-US-00009 TABLE 10 Primary analysis of efficacy endpoints R1033 + R1033 + R1033 + R2477 R2477 R2477 R2477 (10 mg/kg) (1 mg/kg) (3 mg/kg) (10 mg/kg) Analysis Placebo R1033 IV IV IV IV Parameter Visit (n = 12) (n = 6) (n = 6) ( n = 6) (n = 6) (n = 12 ) Thigh Muscle LS Mean 0.37 (0.76) 3.27 (1.07) 0.27 (1.08) 4.96 (1.08) 4.61 (1.07) 5.92 (0.76) Tissue Volume (SE)/Week 4 (cm3), LS Mean 2.9 (1.32)* -0.11 (1.32) 4.59 (1.32)* 4.24 (1.31)* 5.55 (1.07)** Excluding Intra- Diff (SE)/ muscular Adipose week 4 Tissue and Large LS Mean 0.88 (1.05) 4.61 (1.49) 2.85 (1.49) 3.51 (1.49) 6.19 (1.48) 7.73 (1.05) Vessels/% change (SE)/week 8 LS Mean 3.73 (1.82)* 1.97 (1.82) 2.63 (1.82) 5.31 (1.82)* 6.85 (1.48)** Diff (SE)/ week 8 Change LS Mean 1.64 (3.26) 13.95 (4.61) -0.14 (4.63) 20.29 (4.63) 18.77 (4.61) 24.5 (3.26) (SE)/week 4 LS Mean 12.32 (5.65)* -1.78 (5.66) 18.65 (5.67)* 17.14 (5.64)* 22.86 (4.61)** Diff (SE)/ week 4 LS Mean 3.5 (4.13) 20.42 (5.85) 11.22 (5.87) 14.55 (5.88) 25.44 (5.85) 31.95 (4.14) (SE)/week 8 LS Mean 16.91 (7.17)* 7.71 (7.18) 11.05 (7.19) 21.94 (7.16)* 28.45 (5.85)** Diff (SE)/ week 8 Thigh Muscle LS Mean 0.21 (0.78) 3.32 (1.1) -0.2 (1.11) 5 (1.11) 4.49 (1.1) 5.76 (0.78) Volume (cm3), (SE)/week 4 Including LS Mean 3.1 (1.35)* -0.41 (1.36) 4.79 (1.35)* 4.28 (1.35)* 5.54 (1.1)** Intra-muscular Diff (SE)/ Adipose Tissue week 4 and Large LS Mean 0.85 (1.06) 4.14 (1.51) 2.47 (1.52) 3.14 (1.51) 5.99 (1.5) 7.41 (1.06) Vessels/ (SE)/week 8 % change LS Mean 3.29 (1.84) 1.62 (1.85) 2.29 (1.85) 5.14 (1.84)* 6.57 (1.5)** Diff (SE)/ week 8 change LS Mean 0.99 (3.45) 14.76 (4.89) -2.61 (4.94) 21.22 (4.91) 19.35 (4.89) 25.22 (3.45) (SE)/week 4 LS Mean 13.77 (5.99)* -3.6 (6.02) 20.24 (6.01)* 18.36 (5.98)* 24.23 (4.88)** Diff (SE)/ week 4 LS Mean 3.55 (4.42) 19.54 (6.25) 9.26 (6.32) 13.64 (6.29) 26.04 (6.25) 32.27 (4.42) (SE)/week 8 LS Mean 15.99 (7.66)* 5.72 (7.7) 10.09 (7.69) 22.49 (7.65)* 28.72 (6.25)** Diff (SE)/ week 8 Appendicular LS Mean -0.31 (0.74) 1.26 (1.04) 2.15 (1.04) 4.26 (1.04) 4.77 (1.05) 3.7 (0.74) Lean Body Mass (SE)/ (Calculated week 4 by aLBM LS Mean 1.57 (1.28) 2.46 (1.28) 4.57 (1.28)** 5.08 (1.29)** 4.01 (1.04)** Equation) Diff (SE)/ (kg)/% change week 4 LS Mean 0.76 (0.73) 2.65 (1.03) 2.37 (1.03) 3.72 (1.03) 7.15 (1.04) 5.72 (0.73) (SE)/week 8 LS Mean 1.9 (1.27) 1.61 (1.26) 2.96 (1.26)* 6.39 (1.27)** 4.97 (1.03)** Diff (SE)/ week 8 Change LS Mean -0.05 (0.13) 0.23 (0.18) 0.35 (0.18) 0.73 (0.18) 0.84 (0.18) 0.63 (0.13) (SE)/week 4 LS Mean 0.28 (0.22) 0.4 (0.22) 0.78 (0.22)* 0.89 (0.22)** 0.68 (0.18)** Diff (SE)/ week 4 LS Mean 0.11 (0.13) 0.45 (0.19) 0.41 (0.19) 0.64 (0.18) 1.28 (0.19) 1.02 (0.13) (SE) LS Mean 0.33 (0.23) 0.29 (0.23) 0.53 (0.23)* 1.16 (0.23)** 0.91 (0.18)** Diff (SE)/ week 8 Total Lean Mass LS Mean 0.16 (0.56) 0.71 (0.79) 1.53 (0.79) 3.42 (0.79) 3.52 (0.79) 2.84 (0.56) (kg)/% change (SE)/week 4 LS Mean 0.55 (0.97) 1.37 (0.98) 3.26 (0.98)* 3.36 (0.98)* 2.69 (0.79)* Diff (SE)/ week 4 LS Mean 1.31 (0.64) 2.18 (0.89) 1.94 (0.9) 3.16 (0.89) 4.67 (0.89) 4.31 (0.63) (SE)/week 8 LS Mean 0.88 (1.1) 0.63 (1.11) 1.85 (1.1) 3.37 (1.1)* 3 (0.89)* Diff (SE)/ week 8 change LS Mean 0.08 (0.23) 0.25 (0.33) 0.61 (0.33) 1.42 (0.33) 1.44 (0.33) 1.14 (0.23) (SE)/week 4 LS Mean 0.17 (0.41) 0.54 (0.41) 1.34 (0.41)* 1.36 (0.41)* 1.07 (0.33)* Diff (SE)/ week 4 LS Mean 0.53 (0.27) 0.86 (0.37) 0.78 (0.37) 1.27 (0.37) 1.92 (0.37) 1.8 (0.26) (SE)/week 8 LS Mean 0.33 (0.46) 0.25 (0.46) 0.74 (0.46) 1.39 (0.46)* 1.26 (0.37)* Diff (SE)/ week 8 Appendicular LS Mean -0.35 (0.74) 1.2 (1).sup. 2.02 (1) 4.06 (1) 4.51 (1).sup. 3.54 (0.71) Lean Mass (SE)/week 4 (Sum of Lean LS Mean 1.56 (1.25) 2.37 (1.24) 4.42 (1.24)** 4.87 (1.26)** 3.89 (1.02)** Mass of Arms Diff (SE)/ and Legs) week 4 (kg)/% change LS Mean 0.95 (0.72) 2.51 (0.97) 2.22 (0.97) 3.53 (0.97) 6.74 (0.98) 5.48 (0.69) (SE)/week 8 LS Mean 1.56 (1.22) 1.28 (1.21) 2.58 (1.21)* 5.79 (1.23)** 4.53 (0.99)** Diff (SE)/ week 8 change LS Mean -0.06 (0.14) 0.23 (0.18) 0.35 (0.18) 0.73 (0.18) 0.83 (0.19) 0.64 (0.13) (SE)/week 4 LS Mean 0.28 (0.23) 0.41 (0.23) 0.79 (0.23)* 0.89 (0.23)** 0.69 (0.19)** Diff (SE)/ week 4 LS Mean 0.17 (0.14) 0.44 (0.18) 0.4 (0.18) 0.64 (0.18) 1.26 (0.18) 1.02 (0.13) (SE)/week 8 LS Mean 0.27 (0.23) 0.23 (0.23) 0.48 (0.23)* 1.09 (0.23)** 0.86 (0.19)** Diff (SE)/ week 8 Total Fat Mass LS Mean -0.11 (0.81) -1.15 (1.15) 1.32 (1.14) -0.76 (1.14) -1.52 (1.14) -2.58 (0.8) (kg)/% change (SE)/week 4 LS Mean -1.04 (1.41) 1.42 (1.39) -0.65 (1.39) -1.41 (1.39) -2.47 (1.14)* Diff (SE)/ week 4 LS Mean -0.65 (1.12) -2.76 (1.6) -1.34 (1.58) -0.85 (1.58) -2.73 (1.58) -4.57 (1.12) (SE)/week 8 LS Mean -2.11 (1.96) -0.69 (1.93) -0.2 (1.93) -2.08 (1.93) -3.92 (1.58)* Diff (SE)/ week 8 change LS Mean 0.07 (0.21) -0.18 (0.3) 0.29 (0.29) -0.29 (0.29) -0.38 (0.29) -0.6 (0.21) (SE)/week 4 LS Mean -0.25 (0.36) 0.23 (0.36) -0.36 (0.36) -0.45 (0.36) -0.67 (0.29)* Diff (SE)/ week 4 LS Mean -0.04 (0.28) -0.54 (0.4) -0.46 (0.4) -0.34 (0.4) -0.75 (0.4) -1.16 (0.28) (SE)/week 8 LS Mean -0.5 (0.49) -0.42 (0.49) -0.3 (0.49) -0.71 (0.49) -1.12 (0.4)* Diff (SE)/ week 8 Android Fat Mass LS Mean -0.21 (1.3) -1.57 (1.85) 0.44 (1.84) -2.47 (1.84) -1.8 (1.83) -3.94 (1.3) (kg)/% change (SE)/week 4 LS Mean -1.35 (2.26) 0.65 (2.25) -2.25 (2.25) -1.59 (2.25) -3.73 (1.83)* Diff (SE)/ week 4 LS Mean -0.05 (1.69) -2.35 (2.4) -4.67 (2.39) -1.71 (2.39) -2.62 (2.38) -6.65 (1.69) (SE) LS Mean -2.31 (2.93) -4.62 (2.92) -1.66 (2.93) -2.57 (2.92) -6.61 (2.38)* Diff (SE)/ week 8 change LS Mean 0.03 (0.03) 0.01 (0.04) 0 (0.04) -0.05 (0.04) -0.04 (0.04) -0.07 (0.03) (SE)/week 4 LS Mean -0.01 (0.05) -0.02 (0.05) -0.07 (0.05) -0.06 (0.05) -0.1 (0.04)* Diff (SE)/ week 4 LS Mean 0.03 (0.04) -0.01 (0.05) -0.09 (0.05) -0.03 (0.05) -0.07 (0.05) -0.13 (0.04) (SE)/week 8 LS Mean -0.04 (0.06) -0.11 (0.06) -0.06 (0.06) -0.1 (0.06) -0.16 (0.05)* Diff (SE)/ week 8 Thigh Intra- LS Mean -2.9 (3) .sup. .sup. -2 (4.24) -3.44 (4.23) 6.19 (4.28) -2.99 (4.24) 8.25 (2.98) muscular (SE)/week 4 Adipose Tissue LS Mean 0.9 (5.22) -0.54 (5.17) 9.09 (5.27) -0.09 (5.17) 11.15 (4.23)* Volume (cm3)/ Diff (SE)/ % volume week 4 LS Mean -4.19 (4.21) -3.35 (5.96) 0.65 (5.95) 1.14 (6.02) -3.88 (5.96) 7.26 (4.19) (SE)/week 8 LS Mean 0.84 (7.34) 4.84 (7.26) 5.33 (7.41) 0.31 (7.27) 11.45 (5.95) Diff (SE)/ week 8 change LS Mean -0.09 (0.13) -0.07 (0.18) -0.2 (0.18) 0.14 (0.18) -0.12 (0.18) 0.24 (0.13) (SE)/week 4 LS Mean 0.02 (0.22) -0.11 (0.22) 0.23 (0.22) -0.03 (0.22) 0.33 (0.18) Diff (SE)/ week 4 LS Mean -0.17 (0.17) -0.13 (0.24) -0.02 (0.24) -0.08 (0.25) -0.22 (0.24) 0.2 (0.17) (SE)/week 8 LS Mean 0.04 (0.3) 0.15 (0.3) 0.09 (0.3) -0.05 (0.3) 0.37 (0.24) Diff (SE)/ week 8 Sum of Intra- LS Mean 0.98 (1.4) -3.09 (1.98) 1.63 (1.99) -2.09 (2.03) 1.75 (1.99) -1.54 (1.4) muscular and (SE)/week 4 Peri-muscular LS Mean -4.07 (2.43) 0.65 (2.43) -3.07 (2.47) 0.77 (2.43) -2.52 (1.98) Adipose Tissue Diff (SE)/ (IMAT)/% change week 4 LS Mean 0.94 (1.49) -0.12 (2.12) 0.26 (2.13) -0.5 (2.17) -1.63 (2.12) -2.76 (1.49) (SE)/week 8 LS Mean -1.06 (2.6) -0.68 (2.59) -1.44 (2.64) -2.57 (2.59) -3.7 (2.11) Diff (SE)/ week 8 Change LS Mean 1.05 (1.65) -3.45 (2.34) 2.29 (2.35) -3.2 (2.4) 1.93 (2.35) -1.75 (1.65) (SE)/week 4 LS Mean -4.5 (2.87) 1.24 (2.87) -4.25 (2.92) 0.88 (2.87) -2.8 (2.33) Diff (SE)/ week 4 LS Mean 0.69 (1.8) 0.28 (2.56) 0.48 (2.57) -1.73 (2.62) -1.97 (2.57)

-3.04 (1.81) (SE)/week 8 LS Mean -0.4 (3.14) -0.21 (3.13) -2.42 (3.19) -2.66 (3.13) -3.73 (2.55) Diff (SE)/ week 8 Subcutaneous LS Mean -1.27 (1.02) 0.05 (1.45) -0.26 (1.47) -1.67 (1.44) -1.98 (1.44) -1.76 (1.02) Adipose Tissue (SE)/week 4 Volume (cm3)/ LS Mean 1.32 (1.78) 1.01 (1.82) -0.4 (1.76) -0.71 (1.76) -0.49 (1.44) % change Diff (SE)/ week 4 LS Mean 1.31 (1.21) 1.01 (1.71) 1.56 (1.75) -3.73 (1.71) -3.04 (1.7) -1.45 (1.21) (SE)/week 8 LS Mean -0.31 (2.11) 0.25 (2.15) -5.04 (2.09)* -4.36 (2.09)* -2.77 (1.7) Diff (SE)/ week 8 change LS Mean -3.94 (3.83) 0.35 (5.42) -0.74 (5.52) -8.23 (5.39) -3.02 (5.39) -7.29 (3.82) (SE)/week 4 LS Mean 4.29 (6.68) 3.19 (6.8) -4.3 (6.6) 0.92 (6.6) -3.35 (5.39) Diff (SE)/ week 4 LS Mean 4.74 (4.74) 5.7 (6.7) 5.53 (6.83) -17.58 (6.67) -8.75 (6.66) -5.55 (4.73) (SE)/week 8 LS Mean 0.95 (8.26) 0.79 (8.41) -22.33 (8.16)* -13.5 (8.17) -10.3 (6.66) Diff (SE)/ week 8 Sum of Fat LS Mean -0.8 (1.01) -0.59 (1.44) 1.89 (1.42) 0.48 (1.42) -2.87 (1.42) -3.31 (1) Mass of Arms (SE)/week 4 and Legs (kg)/ LS Mean 0.21 (1.78) 2.69 (1.74) 1.28 (1.74) -2.07 (1.74) -2.51 (1.42) % change Diff (SE)/ week 4 LS Mean -0.47 (1.34) -2.44 (1.9) 0.11 (1.87) 0.27 (1.87) -3.51 (1.87) -5.2 (1.32) (SE)/week 8 LS Mean -1.97 (2.35) 0.58 (2.31) 0.74 (2.3) -3.04 (2.3) -4.73 (1.88)* Diff (SE)/ week 8 change LS Mean -0.09 (0.12) -0.06 (0.17) 0.21 (0.16) 0.01 (0.16) -0.33 (0.16) -0.41 (0.11) (SE)/week 4 LS Mean 0.02 (0.2) 0.3 (0.2) 0.09 (0.2) -0.25 (0.2) -0.33 (0.16) Diff (SE)/ week 4 LS Mean -0.05 (0.15) -0.26 (0.22) -0.03 (0.21) -0.07 (0.21) -0.41 (0.21) -0.62 (0.15) (SE)/week 8 LS Mean -0.21 (0.27) 0.02 (0.26) -0.02 (0.26) -0.37 (0.26) -0.58 (0.22)* Diff (SE)/ week 8 Total Bone LS Mean 0.05 (0.25) -0.22 (0.36) -0.48 (0.35) -0.04 (0.36) 0.47 (0.36) 0.55 (0.25) Mineral Density (SE)/week 4 (BMD) Mass LS Mean -0.27 (0.43) -0.53 (0.43) -0.09 (0.44) 0.42 (0.44) 0.5 (0.35) (g/cm2)/% Diff (SE)/ change week 4 LS Mean -0.01 (0.28) -0.47 (0.4) -0.72 (0.4) -0.08 (0.4) -0.22 (0.41) 0.16 (0.28) (SE)/week 8 LS Mean -0.46 (0.49) -0.71 (0.49) -0.07 (0.5) -0.21 (0.5) 0.17 (0.4) Diff (SE)/ week 8 change LS Mean 0 (0.003) -0.002 (0.004) -0.005 (0.004) 0 (0.004) 0.005 (0.004) 0.006 (0.003) (SE)/week 4 LS Mean 0 (0) -0.01 (0) 0 (0) 0 (0) 0.01 (0) Diff (SE)/ week 4 LS Mean 0 (0.003) -0.006 (0.004) -0.008 (0.004) 0 (0.004) -0.002 (0.004) 0.002 (0.003) (SE)/week 8 LS Mean -0.01 (0.01) -0.01 (0.01) 0 (0.01) 0 (0.01) 0 (0) Diff (SE)/ week 8 Total Bone LS Mean 0.1 (0.28) -0.16 (0.4) -0.23 (0.4) 0.25 (0.4) -0.19 (0.4) 0.62 (0.28) Mineral Content (SE)/week 4 (BMC) Mass LS Mean -0.26 (0.49) -0.33 (0.49) 0.15 (0.49) -0.29 (0.49) 0.52 (0.4) (kg)/% change Diff (SE)/ week 4 LS Mean -0.42 (0.26) -0.27 (0.36) -0.48 (0.36) -0.08 (0.36) -0.56 (0.36) 0.65 (0.25) (SE)/week 8 LS Mean 0.16 (0.45) -0.05 (0.44) 0.34 (0.44) -0.14 (0.44) 1.07 (0.36)* Diff (SE)/ week 8 change LS Mean 0.002 (0.006) -0.003 (0.009) -0.005 (0.009) 0.006 (0.009) -0.003 (0.009) 0.014 (0.006) (SE)/week 4 LS Mean -0.01 (0.01) -0.01 (0.01) 0 (0.01) -0.01 (0.01) 0.01 (0.01) Diff (SE)/ week 4 LS Mean -0.009 (0.006) -0.007 (0.008) -0.011 (0.008) -0.001 (0.008) -0.012 (0.008) 0.014 (0.006) (SE)/week 8 LS Mean 0 (0.01) 0 (0.01) 0.01 (0.01) 0 (0.01) 0.02 (0.01)* Diff (SE)/ week 8 T-score for Total LS Mean -18.26 (18.01) -51.19 (25.74) -2.95 (25.45) -6.66 (25.7) -39.76 (25.87) -0.9 (17.96) Body/% change (SE)/week 4 LS Mean -32.93 (31.23) 15.31 (31.1) 11.6 (31.56) -21.5 (31.73) 17.37 (25.43) Diff (SE)/ week 4 LS Mean -20.87 (17.94) -58.54 (25.64) 3.83 (25.35) 5.16 (25.6) 23.12 (25.77) 8.49 (17.89) (SE)/week 8 LS Mean -37.67 (31.11) 24.71 (30.98) 26.04 (31.44) 44 (31.61) 29.36 (25.33) Diff (SE)/ week 8 change LS Mean 0.01 (0.03) -0.03 (0.05) -0.07 (0.05) 0 (0.05) 0.06 (0.05) 0.08 (0.03) (SE)/week 4 LS Mean -0.04 (0.06) -0.07 (0.06) -0.01 (0.06) 0.06 (0.06) 0.07 (0.05) Diff (SE)/ week 4 LS Mean 0 (0.04) -0.07 (0.05) -0.1 (0.05) -0.01 (0.05) -0.03 (0.06) 0.03 (0.04) (SE)/week 8 LS Mean -0.07 (0.07) -0.1 (0.07) -0.01 (0.07) -0.03 (0.07) 0.03 (0.05) Diff (SE)/ week 8 Note: Least-squares (LS) means, standard errors (SE) and p-value taken from ANCOVA. The model includes baseline measurement as covariate and the treatment as fixed factor. *indicated p-value < 0.05; **indicated p-value < 0.001.

[0256] Compared with placebo, R2477+R1033 combination significantly increased thigh muscle volume and total lean mass in medium and high dose groups (FIGS. 8 and 9), significantly increased appendicular lean body mass in all dose groups (FIG. 10) and significantly decreased total fat mass as well as android fat mass in the high dose group (Table 10). R2477+R1033, in the high dose group, significantly increased all of the major measures of muscle volume and lean mass compared with placebo, at both Week 4 and Week 8; the effects at Week 4 were generally less pronounced than at Week 8 (Table 10). At Week 8, the high dose anti-Activin A R2477+anti-GDF8 R1033 group exhibited increased % change in total bone mineral content from placebo, as measured by DXA, while total bone mineral density did not change (Table 10).

Safety

[0257] All treatment emergent adverse events (TEAEs) were mild to moderate in severity except one severe TEAE of `radius fracture` reported by a placebo subject. There were no serious adverse events, no deaths, and no discontinuations due to TEAEs. Headache was the most frequent TEAE in each of the treatment groups, occurring in 58.3% of all study subjects and in 50% of placebo subjects. Muscle spasms, nausea and mouth ulceration were the other frequent TEAEs in REGN2477+REGN1033 groups that occurred in 25% or more of subjects in the combination R2477+R1033 groups; these TEAEs occurred less frequently in the placebo group, but there does not appear to be any clear dose-response relationship. There were no clear signals of bleeding or diarrhea, adverse events that have been associated with blockade of activin receptors. One TEAE of a nosebleed (preferred term of epistaxis) occurred in a R2477+R1033 dose group--it resolved after 9 minutes.

[0258] Review of Potentially Clinically Significant Values (PCSVs) revealed no significant differences between REGN2477+REGN1033 and Placebo in Labs, Vital signs, and ECG that would indicate negative effects of REGN2477+REGN1033. Within laboratory, vital sign and ECG categories, there were 0-2 subjects with PCSVs in the combined REGN2477+REGN1033 dose groups (N of 24); however the percentage of subjects with PCSVs was equal to or lower than that found in the placebo group. There were no treatment-emergent PCSVs related to liver function tests.

CONCLUSIONS

[0259] In healthy postmenopausal women, single intravenous doses of REGN2477+REGN1033 increased thigh muscle volume, total lean mass, and appendicular lean body mass. One surprising finding was the uniformity of the thigh muscle changes: all of the individuals exposed to the combination exhibited an increase in thigh muscle volume, as shown in FIG. 4. In addition, single intravenous doses of REGN2477+REGN1033 decreased total fat, and in particular, android fat mass. Treatment with REGN1033 alone increased thigh muscle volume.

[0260] In general, REGN2477, REGN1033, and REGN2477+REGN1033 in this clinical study were considered to have an acceptable safety profile and were well tolerated. There were no serious adverse events.

[0261] The present invention is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description and the accompanying figures. Such modifications are intended to fall within the scope of the appended claims.

Sequence CWU 1

1

6481366DNAArtificial SequenceSynthetic 1caggtgcagc tggtgcagtc tgggggaggc ttggtccagc ctggggggtc cctgagactc 60tcctgtgcag cctctggatt cacctttaac acctatgcca taagctgggt ccgccaggct 120ccagggaagg ggctggaatg ggtctcaact attactggta gtggttataa cacatactac 180gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacactgtat 240ctacagatga gcagcctgag agccgaggac acggccgtat tttactgtgc gaaagactct 300cggtataact ggaattacgg aatttttgac tactggggcc agggaaccac ggtcaccgtc 360tcctca 3662122PRTArtificial SequenceSynthetic 2Gln Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Thr Tyr 20 25 30Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Thr Ile Thr Gly Ser Gly Tyr Asn Thr Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala Val Phe Tyr Cys 85 90 95Ala Lys Asp Ser Arg Tyr Asn Trp Asn Tyr Gly Ile Phe Asp Tyr Trp 100 105 110Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120324DNAArtificial SequenceSynthetic 3ggattcacct ttaacaccta tgcc 2448PRTArtificial SequenceSynthetic 4Gly Phe Thr Phe Asn Thr Tyr Ala1 5524DNAArtificial SequenceSynthetic 5attactggta gtggttataa caca 2468PRTArtificial SequenceSynthetic 6Ile Thr Gly Ser Gly Tyr Asn Thr1 5745DNAArtificial SequenceSynthetic 7gcgaaagact ctcggtataa ctggaattac ggaatttttg actac 45815PRTArtificial SequenceSynthetic 8Ala Lys Asp Ser Arg Tyr Asn Trp Asn Tyr Gly Ile Phe Asp Tyr1 5 10 159324DNAArtificial SequenceSynthetic 9gacatccaga tgacccagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacca tcagaaacct 120ggccaggctc ccaggctcct catctatggt gtatccacca gggccactgg tatcccagcc 180aggttcagtg gcaatgggtc tgggacagag ttcactctca ccatcagcag cctgcagtct 240gaagattttg cagtttatta ctgtcagcag cataataact ggccgctcac tttcggcgga 300gggaccaagg tggagatcaa acga 32410108PRTArtificial SequenceSynthetic 10Asp Ile Gln Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Leu Ala Trp Tyr His Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Gly Val Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Asn Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln His Asn Asn Trp Pro Leu 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg 100 1051118DNAArtificial SequenceSynthetic 11cagagtgtta gcagcaac 18126PRTArtificial SequenceSynthetic 12Gln Ser Val Ser Ser Asn1 5139DNAArtificial SequenceSynthetic 13ggtgtatcc 9143PRTArtificial SequenceSynthetic 14Gly Val Ser11527DNAArtificial SequenceSynthetic 15cagcagcata ataactggcc gctcact 27169PRTArtificial SequenceSynthetic 16Gln Gln His Asn Asn Trp Pro Leu Thr1 517375DNAArtificial SequenceSynthetic 17caggtgcagc tggtgcagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60acctgcactg tctctggtgg ctccatcagc aatagtaatt actactgggg ctggatccgc 120cagcccccag ggaaggggct ggagtggatt gggactacct attatagtgg gaccacctac 180tacaacccgt ccctcaagag tcgagtcacc atatccgtag acacgtccaa gaaccagttc 240tccctgaagc tgagctctgt gaccgccgca gacacggctg tgtattactg tgcgagagat 300tattatgata gtagtggtta ttattacaac tggttcgatc cctggggcca gggaaccacg 360gtcaccgtct cctca 37518125PRTArtificial SequenceSynthetic 18Gln Val Gln Leu Val Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Asn Ser 20 25 30Asn Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 40 45Trp Ile Gly Thr Thr Tyr Tyr Ser Gly Thr Thr Tyr Tyr Asn Pro Ser 50 55 60Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe65 70 75 80Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 90 95Cys Ala Arg Asp Tyr Tyr Asp Ser Ser Gly Tyr Tyr Tyr Asn Trp Phe 100 105 110Asp Pro Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 1251930DNAArtificial SequenceSynthetic 19ggtggctcca tcagcaatag taattactac 302010PRTArtificial SequenceSynthetic 20Gly Gly Ser Ile Ser Asn Ser Asn Tyr Tyr1 5 102121DNAArtificial SequenceSynthetic 21acctattata gtgggaccac c 21227PRTArtificial SequenceSynthetic 22Thr Tyr Tyr Ser Gly Thr Thr1 52351DNAArtificial SequenceSynthetic 23gcgagagatt attatgatag tagtggttat tattacaact ggttcgatcc c 512417PRTArtificial SequenceSynthetic 24Ala Arg Asp Tyr Tyr Asp Ser Ser Gly Tyr Tyr Tyr Asn Trp Phe Asp1 5 10 15Pro25324DNAArtificial SequenceSynthetic 25gacatccagt tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca ggacattaga aatgatttag gctggtatca gcagaaacca 120gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagaa ttcagtctca cactcagcag cctgcagcct 240gaagattttg caacttattt ctgtctacag catcatattt acccgtggac gttcggccaa 300gggaccaagc tggagatcaa acga 32426108PRTArtificial SequenceSynthetic 26Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Arg Asn Asp 20 25 30Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Leu Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Phe Cys Leu Gln His His Ile Tyr Pro Trp 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 100 1052718DNAArtificial SequenceSynthetic 27caggacatta gaaatgat 18286PRTArtificial SequenceSynthetic 28Gln Asp Ile Arg Asn Asp1 5299DNAArtificial SequenceSynthetic 29gctgcatcc 9303PRTArtificial SequenceSynthetic 30Ala Ala Ser13127DNAArtificial SequenceSynthetic 31ctacagcatc atatttaccc gtggacg 27329PRTArtificial SequenceSynthetic 32Leu Gln His His Ile Tyr Pro Trp Thr1 533375DNAArtificial SequenceSynthetic 33caagttcagc tggtggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60acctgcactg tctatggtgg ctccatcagc agtggtaatt actactgggg ctggatccgc 120cagcccccag ggaagggact ggagtggatt gggactatct attatagtgg aagcgcctac 180tacaacccgt ccctcaagag tcgagtcacc atgtccgtag acacgtccaa gaaccagttc 240tccctgaaac tgagctctgt gaccgccgca gacacggctg tttattactg tgtgagagat 300tactatgata gtagtggtca ttattacaac tggttcgacc cctggggcca gggaaccacg 360gtcaccgtct cctca 37534125PRTArtificial SequenceSynthetic 34Gln Val Gln Leu Val Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Tyr Gly Gly Ser Ile Ser Ser Gly 20 25 30Asn Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 40 45Trp Ile Gly Thr Ile Tyr Tyr Ser Gly Ser Ala Tyr Tyr Asn Pro Ser 50 55 60Leu Lys Ser Arg Val Thr Met Ser Val Asp Thr Ser Lys Asn Gln Phe65 70 75 80Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 90 95Cys Val Arg Asp Tyr Tyr Asp Ser Ser Gly His Tyr Tyr Asn Trp Phe 100 105 110Asp Pro Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 1253530DNAArtificial SequenceSynthetic 35ggtggctcca tcagcagtgg taattactac 303610PRTArtificial SequenceSynthetic 36Gly Gly Ser Ile Ser Ser Gly Asn Tyr Tyr1 5 103721DNAArtificial SequenceSynthetic 37atctattata gtggaagcgc c 21387PRTArtificial SequenceSynthetic 38Ile Tyr Tyr Ser Gly Ser Ala1 53951DNAArtificial SequenceSynthetic 39gtgagagatt actatgatag tagtggtcat tattacaact ggttcgaccc c 514017PRTArtificial SequenceSynthetic 40Val Arg Asp Tyr Tyr Asp Ser Ser Gly His Tyr Tyr Asn Trp Phe Asp1 5 10 15Pro41324DNAArtificial SequenceSynthetic 41gccatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca ggacattaga catgatttag gctggtatca gcagaaacca 120gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 240gaagattttg caacttatta ctgtctacag cataatactt acccgtggac gttcggccaa 300gggaccaagg tggagatcaa acga 32442108PRTArtificial SequenceSynthetic 42Ala Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Arg His Asp 20 25 30Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Thr Tyr Pro Trp 85 90 95Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 1054318DNAArtificial SequenceSynthetic 43caggacatta gacatgat 18446PRTArtificial SequenceSynthetic 44Gln Asp Ile Arg His Asp1 5459DNAArtificial SequenceSynthetic 45gctgcatcc 9463PRTArtificial SequenceSynthetic 46Ala Ala Ser14727DNAArtificial SequenceSynthetic 47ctacagcata atacttaccc gtggacg 27489PRTArtificial SequenceSynthetic 48Leu Gln His Asn Thr Tyr Pro Trp Thr1 549357DNAArtificial SequenceSynthetic 49caggtgcagc tggtgcagtc tgggggaggc ctggtcaagc ctggggggtc cctgagactc 60tcctgtgcag cctctggatt caccttcagt agctatagca tgaactgggt ccgccaggct 120ccagggaagg ggctggagtg ggtctcatcc attagtagta gtagtagtta catatactac 180gcagactcag tgaagggccg attcaccatc tccagagaca acgccaagaa ctcactgtat 240ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc gagagatcgt 300ggatacacct ttggggttga ctactggggc cagggaacca cggtcaccgt ctcctca 35750119PRTArtificial SequenceSynthetic 50Gln Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Ser Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Ser Ile Ser Ser Ser Ser Ser Tyr Ile Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Asp Arg Gly Tyr Thr Phe Gly Val Asp Tyr Trp Gly Gln Gly 100 105 110Thr Thr Val Thr Val Ser Ser 1155124DNAArtificial SequenceSynthetic 51ggattcacct tcagtagcta tagc 24528PRTArtificial SequenceSynthetic 52Gly Phe Thr Phe Ser Ser Tyr Ser1 55324DNAArtificial SequenceSynthetic 53attagtagta gtagtagtta cata 24548PRTArtificial SequenceSynthetic 54Ile Ser Ser Ser Ser Ser Tyr Ile1 55536DNAArtificial SequenceSynthetic 55gcgagagatc gtggatacac ctttggggtt gactac 365612PRTArtificial SequenceSynthetic 56Ala Arg Asp Arg Gly Tyr Thr Phe Gly Val Asp Tyr1 5 1057324DNAArtificial SequenceSynthetic 57gccatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca gggcattaga aatgatttag gctggtatca gcagaaacca 120gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 240gaagattttg caacttatta ctgtctacag cataatagtt acccgtacac ttttggccag 300gggaccaagg tggagatcaa acga 32458108PRTArtificial SequenceSynthetic 58Ala Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Asn Asp 20 25 30Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 1055918DNAArtificial SequenceSynthetic 59cagggcatta gaaatgat 18606PRTArtificial SequenceSynthetic 60Gln Gly Ile Arg Asn Asp1 5619DNAArtificial SequenceSynthetic 61gctgcatcc 9623PRTArtificial SequenceSynthetic 62Ala Ala Ser16327DNAArtificial SequenceSynthetic 63ctacagcata atagttaccc gtacact 27649PRTArtificial SequenceSynthetic 64Leu Gln His Asn Ser Tyr Pro Tyr Thr1 565375DNAArtificial SequenceSynthetic 65caggtgcagc tggtgcagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60acctgcactg tctctggtgg ctccatcatc acttatagtt actactgggg ctggatccgc 120cagcccccag ggaaggggct ggagtggatt gggactatcc atcatagtgg gagcacctac 180tacaacccgt ccctcaagag tcgagtcacc atatccgtag acacgtccaa gaaccagttc 240tccctgacac tgagttctgt gaccgccgca gacacggctg tgtattactg tgcgagagac 300tactatgata gtagtggtta ttattataac tggttcgacc cctggggcca gggaaccatg 360gtcaccgtct cctca 37566125PRTArtificial SequenceSynthetic 66Gln Val Gln Leu Val Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ile Thr Tyr 20 25 30Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 40 45Trp Ile Gly Thr Ile His His Ser Gly Ser Thr Tyr Tyr Asn Pro Ser 50 55 60Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe65 70 75 80Ser Leu Thr Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 90 95Cys Ala Arg Asp Tyr Tyr Asp Ser Ser Gly Tyr Tyr Tyr Asn Trp Phe 100 105 110Asp Pro Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 1256730DNAArtificial SequenceSynthetic 67ggtggctcca tcatcactta tagttactac 306810PRTArtificial SequenceSynthetic 68Gly Gly Ser Ile Ile Thr Tyr Ser Tyr Tyr1 5 106921DNAArtificial SequenceSynthetic 69atccatcata gtgggagcac c 21707PRTArtificial SequenceSynthetic 70Ile His His Ser Gly Ser Thr1 57151DNAArtificial SequenceSynthetic

71gcgagagact actatgatag tagtggttat tattataact ggttcgaccc c 517217PRTArtificial SequenceSynthetic 72Ala Arg Asp Tyr Tyr Asp Ser Ser Gly Tyr Tyr Tyr Asn Trp Phe Asp1 5 10 15Pro73324DNAArtificial SequenceSynthetic 73gccatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca gggcattaga aatgatttag gctggtatca gcagaaacca 120gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 240gaagattttg caacttatta ctgtctacag cataatagtt ccccgtggac gttcggccaa 300gggaccaagg tggagatcaa acga 32474108PRTArtificial SequenceSynthetic 74Ala Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Asn Asp 20 25 30Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Ser Ser Pro Trp 85 90 95Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 1057518DNAArtificial SequenceSynthetic 75cagggcatta gaaatgat 18766PRTArtificial SequenceSynthetic 76Gln Gly Ile Arg Asn Asp1 5779DNAArtificial SequenceSynthetic 77gctgcatcc 9783PRTArtificial SequenceSynthetic 78Ala Ala Ser17927DNAArtificial SequenceSynthetic 79ctacagcata atagttcccc gtggacg 27809PRTArtificial SequenceSynthetic 80Leu Gln His Asn Ser Ser Pro Trp Thr1 581369DNAArtificial SequenceSynthetic 81caggtgcact tgaaggagtc tggtcctacg ctggtgaaac ccacacagac cctcacgctg 60acctgcacct tctctgggtt ctcactcagc actagtggag tgggtgtggg ctggatccgt 120cagcccccag gaaaggccct ggagtggctt gcactcattt attggaatga tgataagcgc 180tacagcccat ctctgaagag caggctcacc atcaccaagg acacctccaa aaaccaggtg 240gtccttacaa tgaccaacat ggaccctgtg gacacagcca catattactg tacacacacc 300tcccgttata actggcacta cggcttcctt gactactggg gccagggaac cacggtcacc 360gtctcctca 36982123PRTArtificial SequenceSynthetic 82Gln Val His Leu Lys Glu Ser Gly Pro Thr Leu Val Lys Pro Thr Gln1 5 10 15Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ser 20 25 30Gly Val Gly Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45Trp Leu Ala Leu Ile Tyr Trp Asn Asp Asp Lys Arg Tyr Ser Pro Ser 50 55 60Leu Lys Ser Arg Leu Thr Ile Thr Lys Asp Thr Ser Lys Asn Gln Val65 70 75 80Val Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr 85 90 95Cys Thr His Thr Ser Arg Tyr Asn Trp His Tyr Gly Phe Leu Asp Tyr 100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 1208330DNAArtificial SequenceSynthetic 83gggttctcac tcagcactag tggagtgggt 308410PRTArtificial SequenceSynthetic 84Gly Phe Ser Leu Ser Thr Ser Gly Val Gly1 5 108521DNAArtificial SequenceSynthetic 85atttattgga atgatgataa g 21867PRTArtificial SequenceSynthetic 86Ile Tyr Trp Asn Asp Asp Lys1 58745DNAArtificial SequenceSynthetic 87acacacacct cccgttataa ctggcactac ggcttccttg actac 458815PRTArtificial SequenceSynthetic 88Thr His Thr Ser Arg Tyr Asn Trp His Tyr Gly Phe Leu Asp Tyr1 5 10 1589324DNAArtificial SequenceSynthetic 89gccatccaga tgacccagtc tccatcctca ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgtc gggcgagtca gggcattagc aattatttag cctggtttca gcagaaacca 120gggaaagccc ctaagtccct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180aagttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240gaagattttg caacttatta ctgccaacaa tataatagtt acccgctcac tttcggcgga 300gggaccaagg tggaaatcaa acga 32490108PRTArtificial SequenceSynthetic 90Ala Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Asn Tyr 20 25 30Leu Ala Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Ser Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Lys Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Leu 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg 100 1059118DNAArtificial SequenceSynthetic 91cagggcatta gcaattat 18926PRTArtificial SequenceSynthetic 92Gln Gly Ile Ser Asn Tyr1 5939DNAArtificial SequenceSynthetic 93gctgcatcc 9943PRTArtificial SequenceSynthetic 94Ala Ala Ser19527DNAArtificial SequenceSynthetic 95caacaatata atagttaccc gctcact 27969PRTArtificial SequenceSynthetic 96Gln Gln Tyr Asn Ser Tyr Pro Leu Thr1 597375DNAArtificial SequenceSynthetic 97gaggtgcagc tggtgcagtt gggcccagga ctggtgaagc cttcggagac cctgtccctc 60acctgcactg tctctggtgg ctccatcagc aatagtaatt actactgggg ctggatccgc 120cagcccccag ggaaggggct ggagtggatt gggactacct attatagtgg gaccacctac 180tacaacccgt ccctcaagag tcgagtcacc atatccgtag acacgtccaa gaaccagttc 240tccctgaagc tgagctctgt gaccgccgca gacacggctg tgtattactg tgcgagagat 300tattatgata gtagtggtta ttattacaac tggttcgatc cctggggcca gggaaccacg 360gtcaccgtct cctca 37598125PRTArtificial SequenceSynthetic 98Glu Val Gln Leu Val Gln Leu Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Asn Ser 20 25 30Asn Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 40 45Trp Ile Gly Thr Thr Tyr Tyr Ser Gly Thr Thr Tyr Tyr Asn Pro Ser 50 55 60Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe65 70 75 80Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 90 95Cys Ala Arg Asp Tyr Tyr Asp Ser Ser Gly Tyr Tyr Tyr Asn Trp Phe 100 105 110Asp Pro Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 1259930DNAArtificial SequenceSynthetic 99ggtggctcca tcagcaatag taattactac 3010010PRTArtificial SequenceSynthetic 100Gly Gly Ser Ile Ser Asn Ser Asn Tyr Tyr1 5 1010121DNAArtificial SequenceSynthetic 101acctattata gtgggaccac c 211027PRTArtificial SequenceSynthetic 102Thr Tyr Tyr Ser Gly Thr Thr1 510351DNAArtificial SequenceSynthetic 103gcgagagatt attatgatag tagtggttat tattacaact ggttcgatcc c 5110417PRTArtificial SequenceSynthetic 104Ala Arg Asp Tyr Tyr Asp Ser Ser Gly Tyr Tyr Tyr Asn Trp Phe Asp1 5 10 15Pro105324DNAArtificial SequenceSynthetic 105gccatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca ggacattaga aatgatttag gctggtatca gcagaaacca 120gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagaa ttcagtctca cactcagcag cctgcagcct 240gaagattttg caacttattt ctgtctacag catcatattt acccgtggac gttcggccaa 300gggaccaagg tggagatcaa acga 324106108PRTArtificial SequenceSynthetic 106Ala Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Arg Asn Asp 20 25 30Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Leu Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Phe Cys Leu Gln His His Ile Tyr Pro Trp 85 90 95Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 10510718DNAArtificial SequenceSynthetic 107caggacatta gaaatgat 181086PRTArtificial SequenceSynthetic 108Gln Asp Ile Arg Asn Asp1 51099DNAArtificial SequenceSynthetic 109gctgcatcc 91103PRTArtificial SequenceSynthetic 110Ala Ala Ser111127DNAArtificial SequenceSynthetic 111ctacagcatc atatttaccc gtggacg 271129PRTArtificial SequenceSynthetic 112Leu Gln His His Ile Tyr Pro Trp Thr1 5113355DNAArtificial SequenceSynthetic 113caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cctctggatt caccttcagt cgctatggca ttcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggctgtt atatcttatg atggaagtga tgaatactat 180gtagactccg tgaagggccg attcagcatc tcccgagaca attccaagaa cacgctttat 240ctacaaatga acagtctgag gcctgcggac tcggctgttt attactgtgt gaaaggagat 300ctggaacttg gttttgacta ctggggccag ggaaccctgg tcaccgtctc ctcaa 355114118PRTArtificial SequenceSynthetic 114Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr 20 25 30Gly Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser Tyr Asp Gly Ser Asp Glu Tyr Tyr Val Asp Ser Val 50 55 60Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Pro Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95Val Lys Gly Asp Leu Glu Leu Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser 11511524DNAArtificial SequenceSynthetic 115ggattcacct tcagtcgcta tggc 241168PRTArtificial SequenceSynthetic 116Gly Phe Thr Phe Ser Arg Tyr Gly1 511724DNAArtificial SequenceSynthetic 117atatcttatg atggaagtga tgaa 241188PRTArtificial SequenceSynthetic 118Ile Ser Tyr Asp Gly Ser Asp Glu1 511933DNAArtificial SequenceSynthetic 119gtgaaaggag atctggaact tggttttgac tac 3312011PRTArtificial SequenceSynthetic 120Val Lys Gly Asp Leu Glu Leu Gly Phe Asp Tyr1 5 10121337DNAArtificial SequenceSynthetic 121gatattgtga tgactcaggc tgcaccctct atacctgtca ttccaggaga gtcagtatcc 60atgtcctgca ggtctagtaa gagtctcctg tacagtaatg gacatactta cgtgtattgg 120tttgtgcaga ggccaggcca gtctcctcag ctcctgatat atcggatgtc caaccttgcc 180tcaggagtcc cagacaggtt cagtggcagt gggtcaggaa ctgctttcac actgagaatc 240agtagagtgg aggctgagga tgtgggtgtt tattactgta tgcaaaatct agaatttccg 300ctcacgttcg gtgctgggac caagctggag ctgaaac 337122112PRTArtificial SequenceSynthetic 122Asp Ile Val Met Thr Gln Ala Ala Pro Ser Ile Pro Val Ile Pro Gly1 5 10 15Glu Ser Val Ser Met Ser Cys Arg Ser Ser Lys Ser Leu Leu Tyr Ser 20 25 30Asn Gly His Thr Tyr Val Tyr Trp Phe Val Gln Arg Pro Gly Gln Ser 35 40 45Pro Gln Leu Leu Ile Tyr Arg Met Ser Asn Leu Ala Ser Gly Val Pro 50 55 60Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Ala Phe Thr Leu Arg Ile65 70 75 80Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Asn 85 90 95Leu Glu Phe Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 11012333DNAArtificial SequenceSynthetic 123aagagtctcc tgtacagtaa tggacatact tac 3312411PRTArtificial SequenceSynthetic 124Lys Ser Leu Leu Tyr Ser Asn Gly His Thr Tyr1 5 101259DNAArtificial SequenceSynthetic 125cggatgtcc 91263PRTArtificial SequenceSynthetic 126Arg Met Ser112727DNAArtificial SequenceSynthetic 127atgcaaaatc tagaatttcc gctcacg 271289PRTArtificial SequenceSynthetic 128Met Gln Asn Leu Glu Phe Pro Leu Thr1 5129355DNAArtificial SequenceSynthetic 129caggtgcagc tggtggaggc ggggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cctctggatt caccttcagt cgctatggca ttcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggctgtt atatcttatg atggaactga tgaatactat 180gcagactccg tgaagggccg attcaccatc tcccgagaca attccaagaa cacgctttat 240ctacaaatga acagtctgag acctgcggac tcggctgtat attactgtgc gaaaggagat 300ctggaacttg gttttgacta ctggggccag ggaaccctgg tcaccgtctc ctcaa 355130118PRTArtificial SequenceSynthetic 130Gln Val Gln Leu Val Glu Ala Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr 20 25 30Gly Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser Tyr Asp Gly Thr Asp Glu Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Pro Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95Ala Lys Gly Asp Leu Glu Leu Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser 11513124DNAArtificial SequenceSynthetic 131ggattcacct tcagtcgcta tggc 241328PRTArtificial SequenceSynthetic 132Gly Phe Thr Phe Ser Arg Tyr Gly1 513324DNAArtificial SequenceSynthetic 133atatcttatg atggaactga tgaa 241348PRTArtificial SequenceSynthetic 134Ile Ser Tyr Asp Gly Thr Asp Glu1 513533DNAArtificial SequenceSynthetic 135gcgaaaggag atctggaact tggttttgac tac 3313611PRTArtificial SequenceSynthetic 136Ala Lys Gly Asp Leu Glu Leu Gly Phe Asp Tyr1 5 10137322DNAArtificial SequenceSynthetic 137gccatccggt tgacccagtc tccatcttcc gtgtctgcat ctgtgggaga cagagtcacc 60atcacttgtc gggcgagtca ggatattagt atttggttag cctggtatca gcagagtcca 120gggaaagccc ctaaactcct gatcaatgtt gcatcccgtt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcaacgg tctgcagcct 240gaagattttg taacttacta ttgtcaacag gctaacagtt tcccgatcac cttcggccaa 300gggacacgac tggcgaccaa ac 322138107PRTArtificial SequenceSynthetic 138Ala Ile Arg Leu Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Ile Trp 20 25 30Leu Ala Trp Tyr Gln Gln Ser Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Asn Val Ala Ser Arg Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn Gly Leu Gln Pro65 70 75 80Glu Asp Phe Val Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Ile 85 90 95Thr Phe Gly Gln Gly Thr Arg Leu Ala Thr Lys 100 10513918DNAArtificial SequenceSynthetic 139caggatatta gtatttgg 181406PRTArtificial SequenceSynthetic 140Gln Asp Ile Ser Ile Trp1 51419DNAArtificial SequenceSynthetic 141gttgcatcc 91423PRTArtificial SequenceSynthetic 142Val Ala Ser114327DNAArtificial SequenceSynthetic

143caacaggcta acagtttccc gatcacc 271449PRTArtificial SequenceSynthetic 144Gln Gln Ala Asn Ser Phe Pro Ile Thr1 5145358DNAArtificial SequenceSynthetic 145caggtgcagc tggtgcagtc tgggggaggc ctggtcaagc ctggggggtc cctgagactc 60tcctgtgcag cctctggatt catcttcaat acctatacca tgaattgggt ccgccaggct 120ccagggaagg ggctggagtg ggtctcatcc atcactagtc gtggtactta tatattctac 180tcagactcac ttaagggccg attcaccatt tccagagaca acgccaataa ctcactgttt 240ctgcaaatga acagcctgag agtcgaagac acggctgttt attactgttc gagagatcgt 300ggatacacct ttggtcctga ctactggggc cagggaaccc tggtcaccgt ctcttcag 358146119PRTArtificial SequenceSynthetic 146Gln Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ile Phe Asn Thr Tyr 20 25 30Thr Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Ser Ile Thr Ser Arg Gly Thr Tyr Ile Phe Tyr Ser Asp Ser Leu 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Asn Asn Ser Leu Phe65 70 75 80Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ser Arg Asp Arg Gly Tyr Thr Phe Gly Pro Asp Tyr Trp Gly Gln Gly 100 105 110Thr Leu Val Thr Val Ser Ser 11514724DNAArtificial SequenceSynthetic 147ggattcatct tcaataccta tacc 241488PRTArtificial SequenceSynthetic 148Gly Phe Ile Phe Asn Thr Tyr Thr1 514924DNAArtificial SequenceSynthetic 149atcactagtc gtggtactta tata 241508PRTArtificial SequenceSynthetic 150Ile Thr Ser Arg Gly Thr Tyr Ile1 515136DNAArtificial SequenceSynthetic 151tcgagagatc gtggatacac ctttggtcct gactac 3615212PRTArtificial SequenceSynthetic 152Ser Arg Asp Arg Gly Tyr Thr Phe Gly Pro Asp Tyr1 5 10153322DNAArtificial SequenceSynthetic 153gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca ggacattaga aatgatttag gctggtatca gcagaaacca 120gggaaagccc ctaagggcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 240gaagattttg caacttatca ctgtctacat tatgattttc atcctcggac gttcggccaa 300gggaccaagg tggaaatcaa gc 322154107PRTArtificial SequenceSynthetic 154Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Arg Asn Asp 20 25 30Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Gly Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr His Cys Leu His Tyr Asp Phe His Pro Arg 85 90 95Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 10515518DNAArtificial SequenceSynthetic 155caggacatta gaaatgat 181566PRTArtificial SequenceSynthetic 156Gln Asp Ile Arg Asn Asp1 51579DNAArtificial SequenceSynthetic 157gctgcatcc 91583PRTArtificial SequenceSynthetic 158Ala Ala Ser115927DNAArtificial SequenceSynthetic 159ctacattatg attttcatcc tcggacg 271609PRTArtificial SequenceSynthetic 160Leu His Tyr Asp Phe His Pro Arg Thr1 5161391DNAArtificial SequenceSynthetic 161caggtgcaac tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccttcagt agctatgcca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt atatggtatg atggaactaa ttattactat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc gagagatccc 300ctatattacg atattttgac tggttattcc cccgactact actacggtat ggacgtctgg 360ggccaaggga ccacggtcac cgtctcctca g 391162130PRTArtificial SequenceSynthetic 162Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Asp Gly Thr Asn Tyr Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Asp Pro Leu Tyr Tyr Asp Ile Leu Thr Gly Tyr Ser Pro Asp 100 105 110Tyr Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val 115 120 125Ser Ser 13016324DNAArtificial SequenceSynthetic 163ggattcacct tcagtagcta tgcc 241648PRTArtificial SequenceSynthetic 164Gly Phe Thr Phe Ser Ser Tyr Ala1 516524DNAArtificial SequenceSynthetic 165atatggtatg atggaactaa ttat 241668PRTArtificial SequenceSynthetic 166Ile Trp Tyr Asp Gly Thr Asn Tyr1 516769DNAArtificial SequenceSynthetic 167gcgagagatc ccctatatta cgatattttg actggttatt cccccgacta ctactacggt 60atggacgtc 6916823PRTArtificial SequenceSynthetic 168Ala Arg Asp Pro Leu Tyr Tyr Asp Ile Leu Thr Gly Tyr Ser Pro Asp1 5 10 15Tyr Tyr Tyr Gly Met Asp Val 20169322DNAArtificial SequenceSynthetic 169gaaatagtga tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60ctctcctgca gggccagtca gacttttagc agcaacttag cctggtacca gcagaaacct 120ggccaggctc ccaggctcct catctatggt gcatccacca gggccactgg tatcccagcc 180aggttcagtg gcagtgggtc tgggacagag ttcactctca ccatcagcag cctgcagtct 240gaagattttg cagtttatta ctgtcagcag tataataagt ggccgctcac tttcggcgga 300gggaccaagg tggagatcaa ac 322170107PRTArtificial SequenceSynthetic 170Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Thr Phe Ser Ser Asn 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Lys Trp Pro Leu 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 10517118DNAArtificial SequenceSynthetic 171cagactttta gcagcaac 181726PRTArtificial SequenceSynthetic 172Gln Thr Phe Ser Ser Asn1 51739DNAArtificial SequenceSynthetic 173ggtgcatcc 91743PRTArtificial SequenceSynthetic 174Gly Ala Ser117527DNAArtificial SequenceSynthetic 175cagcagtata ataagtggcc gctcact 271769PRTArtificial SequenceSynthetic 176Gln Gln Tyr Asn Lys Trp Pro Leu Thr1 5177367DNAArtificial SequenceSynthetic 177gaggtgcagc tgttggagtc tgggggaggc ttggtccagc ctggggagtc cctgagactc 60tcctgtgcag cctctggatt cacctttagc agctatgcca tgaactgggt ccgccaggct 120ccagggaagg ggctggagtg ggtctcaact atcagtggta gtggtggtta tatatactac 180gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaagac acggccgtat atttctgtgc gaaagattcc 300aggtataact ggaactacgg caattttgac tactggggcc agggaaccct ggtcaccgtc 360tcctcag 367178122PRTArtificial SequenceSynthetic 178Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Glu1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Thr Ile Ser Gly Ser Gly Gly Tyr Ile Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Phe Cys 85 90 95Ala Lys Asp Ser Arg Tyr Asn Trp Asn Tyr Gly Asn Phe Asp Tyr Trp 100 105 110Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 12017924DNAArtificial SequenceSynthetic 179ggattcacct ttagcagcta tgcc 241808PRTArtificial SequenceSynthetic 180Gly Phe Thr Phe Ser Ser Tyr Ala1 518124DNAArtificial SequenceSynthetic 181atcagtggta gtggtggtta tata 241828PRTArtificial SequenceSynthetic 182Ile Ser Gly Ser Gly Gly Tyr Ile1 518345DNAArtificial SequenceSynthetic 183gcgaaagatt ccaggtataa ctggaactac ggcaattttg actac 4518415PRTArtificial SequenceSynthetic 184Ala Lys Asp Ser Arg Tyr Asn Trp Asn Tyr Gly Asn Phe Asp Tyr1 5 10 15185322DNAArtificial SequenceSynthetic 185gaaatagtga tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60ctctcctgca gggccagtca gaatgttagc agcaacttag cctggaacaa gcagaaacct 120ggccaggctc ccagactcct catctatgct acatccacca gggccactgg tgtcccagcc 180aggttcagtg ccagtgggtc tgggacagac ttcgctctca ccatcaacag cctgcagtct 240gaagattttg cagtttatta ctgtcagcag tataataact ggcctctcac tttcggcgga 300gggaccaagg tggagatcaa ac 322186107PRTArtificial SequenceSynthetic 186Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asn Val Ser Ser Asn 20 25 30Leu Ala Trp Asn Lys Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Ala Thr Ser Thr Arg Ala Thr Gly Val Pro Ala Arg Phe Ser Ala 50 55 60Ser Gly Ser Gly Thr Asp Phe Ala Leu Thr Ile Asn Ser Leu Gln Ser65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Leu 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 10518718DNAArtificial SequenceSynthetic 187cagaatgtta gcagcaac 181886PRTArtificial SequenceSynthetic 188Gln Asn Val Ser Ser Asn1 51899DNAArtificial SequenceSynthetic 189gctacatcc 91903PRTArtificial SequenceSynthetic 190Ala Thr Ser119127DNAArtificial SequenceSynthetic 191cagcagtata ataactggcc tctcact 271929PRTArtificial SequenceSynthetic 192Gln Gln Tyr Asn Asn Trp Pro Leu Thr1 5193367DNAArtificial SequenceSynthetic 193gaggtgcaac tgttggaatc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgtag cctctcgatt caccttcagc agcaatgcca tgagttgggt ccgccaggct 120ccagggacgg ggctggagtg ggtctcagct attactggta gtggtagtag gacatactac 180gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacggtgtat 240ctgcaaatga acagcctgag aggcgaggac acggccgtat attactgtgc gaaagatcaa 300gggggtacct ggaactacgg agattttgac tactggggcc agggaaccct ggtcaccgtc 360tcctcag 367194122PRTArtificial SequenceSynthetic 194Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Val Ala Ser Arg Phe Thr Phe Ser Ser Asn 20 25 30Ala Met Ser Trp Val Arg Gln Ala Pro Gly Thr Gly Leu Glu Trp Val 35 40 45Ser Ala Ile Thr Gly Ser Gly Ser Arg Thr Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Gly Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Lys Asp Gln Gly Gly Thr Trp Asn Tyr Gly Asp Phe Asp Tyr Trp 100 105 110Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 12019524DNAArtificial SequenceSynthetic 195cgattcacct tcagcagcaa tgcc 241968PRTArtificial SequenceSynthetic 196Arg Phe Thr Phe Ser Ser Asn Ala1 519724DNAArtificial SequenceSynthetic 197attactggta gtggtagtag gaca 241988PRTArtificial SequenceSynthetic 198Ile Thr Gly Ser Gly Ser Arg Thr1 519945DNAArtificial SequenceSynthetic 199gcgaaagatc aagggggtac ctggaactac ggagattttg actac 4520015PRTArtificial SequenceSynthetic 200Ala Lys Asp Gln Gly Gly Thr Trp Asn Tyr Gly Asp Phe Asp Tyr1 5 10 15201322DNAArtificial SequenceSynthetic 201gaaatagtga tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacca gcagaaacct 120ggccaggctc ccaggctcct catctatggt gcatccacca gggccactgg tctcccagcc 180aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctgcagtct 240gaagattttg cagtttatta ctgtcagcag tataataact ggcctctcac tttcggcgga 300gggaccaagg tggagatcaa gc 322202107PRTArtificial SequenceSynthetic 202Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Gly Ala Ser Thr Arg Ala Thr Gly Leu Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Leu 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 10520318DNAArtificial SequenceSynthetic 203cagagtgtta gcagcaac 182046PRTArtificial SequenceSynthetic 204Gln Ser Val Ser Ser Asn1 52059DNAArtificial SequenceSynthetic 205ggtgcatcc 92063PRTArtificial SequenceSynthetic 206Gly Ala Ser120727DNAArtificial SequenceSynthetic 207cagcagtata ataactggcc tctcact 272089PRTArtificial SequenceSynthetic 208Gln Gln Tyr Asn Asn Trp Pro Leu Thr1 5209373DNAArtificial SequenceSynthetic 209caggtgcagc tggtggagtc tgggggagac gtggtccagc ctgggaggtc cctgagactc 60tcctgtacag cctctggatt caccttcagt agttatggca tgcactgggt ccgccaggct 120ccaggcaggg ggctggagtg ggtggcagtt atatcatttg atggaaaaaa taaatactat 180gcagactccg tgaagggccg attcaccgtc tccagagaca attccaagaa cacgctgttt 240ctgcaaatga acagcctgag agctgaggac acggctctat attactgtgc gaaaaggata 300gcagcaactg gttactacta cttctacggt ttggacgtct ggggccaagg gaccacggtc 360accgtctcct cag 373210124PRTArtificial Sequence`Synthetic 210Gln Val Gln Leu Val Glu Ser Gly Gly Asp Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Arg Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser Phe Asp Gly Lys Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95Ala Lys Arg Ile Ala Ala Thr Gly Tyr Tyr Tyr Phe Tyr Gly Leu Asp 100 105 110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 12021124DNAArtificial SequenceSynthetic 211ggattcacct tcagtagtta tggc

242128PRTArtificial SequenceSynthetic 212Gly Phe Thr Phe Ser Ser Tyr Gly1 521324DNAArtificial SequenceSynthetic 213atatcatttg atggaaaaaa taaa 242148PRTArtificial SequenceSynthetic 214Ile Ser Phe Asp Gly Lys Asn Lys1 521551DNAArtificial SequenceSynthetic 215gcgaaaagga tagcagcaac tggttactac tacttctacg gtttggacgt c 5121617PRTArtificial SequenceSynthetic 216Ala Lys Arg Ile Ala Ala Thr Gly Tyr Tyr Tyr Phe Tyr Gly Leu Asp1 5 10 15Val217322DNAArtificial SequenceSynthetic 217gaaataatga tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagaggcacc 60ctctcctgca gggccagtca gagtgttagt agcaacttag cctggtacca gcagaaacct 120ggccaggctc ccaggctcct catctatggt gcatccacca gggccagtgg tatcccagcc 180aggttcagtg gcagtgggtc tgggacagag ttcactctca ccgtcagcag cctgcagtct 240gaagattttg cagtttatta ctgtcagcag tataataact ggccgctcac tttcggcgga 300gggaccaagg tggagatcaa ac 322218107PRTArtificial SequenceSynthetic 218Glu Ile Met Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly1 5 10 15Glu Arg Gly Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Gly Ala Ser Thr Arg Ala Ser Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Val Ser Ser Leu Gln Ser65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Leu 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 10521918DNAArtificial SequenceSynthetic 219cagagtgtta gtagcaac 182206PRTArtificial SequenceSynthetic 220Gln Ser Val Ser Ser Asn1 52219DNAArtificial SequenceSynthetic 221ggtgcatcc 92223PRTArtificial SequenceSynthetic 222Gly Ala Ser122327DNAArtificial SequenceSynthetic 223cagcagtata ataactggcc gctcact 272249PRTArtificial SequenceSynthetic 224Gln Gln Tyr Asn Asn Trp Pro Leu Thr1 5225367DNAArtificial SequenceSynthetic 225gaggttcagc tgttggagtc tgggggaggc ttggtacagc cgggggggtc cctgagactc 60tcctgtgcag cctctggatt cacctctatc acctatgcca tgagctgggt ccgccaggct 120ccagggaagg ggctggagtg ggtctcagct attagtgtta gtggtactaa tacatactac 180gcagactccg tgaagggccg gttcaccatc tccagagaca agtccaagaa catgctgtat 240ctgcaaatga acagcctgag agccgaggac acggccgtgt attactgtgc gaaagatctc 300ctacataact ggaaatacgg gacttttgat atctggggcc aagggacaat ggtcaccgtc 360tcttcag 367226122PRTArtificial SequenceSynthetic 226Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Ser Ile Thr Tyr 20 25 30Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Ala Ile Ser Val Ser Gly Thr Asn Thr Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Lys Ser Lys Asn Met Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Lys Asp Leu Leu His Asn Trp Lys Tyr Gly Thr Phe Asp Ile Trp 100 105 110Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 12022724DNAArtificial SequenceSynthetic 227ggattcacct ctatcaccta tgcc 242288PRTArtificial SequenceSynthetic 228Gly Phe Thr Ser Ile Thr Tyr Ala1 522924DNAArtificial SequenceSynthetic 229attagtgtta gtggtactaa taca 242308PRTArtificial SequenceSynthetic 230Ile Ser Val Ser Gly Thr Asn Thr1 523145DNAArtificial SequenceSynthetic 231gcgaaagatc tcctacataa ctggaaatac gggacttttg atatc 4523215PRTArtificial SequenceSynthetic 232Ala Lys Asp Leu Leu His Asn Trp Lys Tyr Gly Thr Phe Asp Ile1 5 10 15233322DNAArtificial SequenceSynthetic 233gaaatagtga tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60ctctcctgca gggccagtca gagtgttgac agcaacttag tctggtacca acaaaaacct 120ggccaggttc ccaggctcct catctatggt gcatccacca gggccactgg tatcccagcc 180aggttcagtg gcagtgggtc tgggacagag ttcactctca ccatcagcag cctgcagtct 240gaagattttg cagtttatta ctgtcagcag tataataagt ggccgctcac tttcggcgga 300gggaccaagg tggagatcaa ac 322234107PRTArtificial SequenceSynthetic 234Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Asp Ser Asn 20 25 30Leu Val Trp Tyr Gln Gln Lys Pro Gly Gln Val Pro Arg Leu Leu Ile 35 40 45Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Lys Trp Pro Leu 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 10523518DNAArtificial SequenceSynthetic 235cagagtgttg acagcaac 182366PRTArtificial SequenceSynthetic 236Gln Ser Val Asp Ser Asn1 52379DNAArtificial SequenceSynthetic 237ggtgcatcc 92383PRTArtificial SequenceSynthetic 238Gly Ala Ser123927DNAArtificial SequenceSynthetic 239cagcagtata ataagtggcc gctcact 272409PRTArtificial SequenceSynthetic 240Gln Gln Tyr Asn Lys Trp Pro Leu Thr1 5241367DNAArtificial SequenceSynthetic 241gaggttcagc tgttggagtc tgggggaggc ttggtacagc cgggggggtc cctgagactc 60tcctgtgcag cctctggatt cacctctatc acctatgcca tgagctgggt ccgccaggct 120ccagggaagg ggctggagtg ggtctcagct attagtgtta gtggtactaa tacatactac 180gcagactccg tgaagggccg gttcaccatc tccagagaca agtccaagaa catgctgtat 240ctgcaaatga acagcctgag agccgaggac acggccgtgt attactgtgc gaaagatctc 300ctacataact ggaaatacgg gacttttgat atctggggcc aagggacaat ggtcaccgtc 360tcttcag 367242122PRTArtificial SequenceSynthetic 242Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Ser Ile Thr Tyr 20 25 30Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Ala Ile Ser Val Ser Gly Thr Asn Thr Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Lys Ser Lys Asn Met Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Lys Asp Leu Leu His Asn Trp Lys Tyr Gly Thr Phe Asp Ile Trp 100 105 110Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 12024324DNAArtificial SequenceSynthetic 243ggattcacct ctatcaccta tgcc 242448PRTArtificial SequenceSynthetic 244Gly Phe Thr Ser Ile Thr Tyr Ala1 524524DNAArtificial SequenceSynthetic 245attagtgtta gtggtactaa taca 242468PRTArtificial SequenceSynthetic 246Ile Ser Val Ser Gly Thr Asn Thr1 524745DNAArtificial SequenceSynthetic 247gcgaaagatc tcctacataa ctggaaatac gggacttttg atatc 4524815PRTArtificial SequenceSynthetic 248Ala Lys Asp Leu Leu His Asn Trp Lys Tyr Gly Thr Phe Asp Ile1 5 10 15249325DNAArtificial SequenceSynthetic 249gaaatagtga tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60ctctcctgca gggccagtca gagtgttagc agcaacttag cctggaacca gcagaaacct 120ggccaggctc ccaggctcct catctatggt gcatccacca gggccacggg tatcccagcc 180aggttcagtg gcagtgggtc tgggacagag ttcactctca ccatcagcag cctgcagtct 240gaagattttg cagtttatta ctgtcagcag tataacaact ggcccatgta cacttttggc 300caggggacca agctggagat caaac 325250108PRTArtificial SequenceSynthetic 250Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Leu Ala Trp Asn Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Met 85 90 95Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 10525118DNAArtificial SequenceSynthetic 251cagagtgtta gcagcaac 182526PRTArtificial SequenceSynthetic 252Gln Ser Val Ser Ser Asn1 52539DNAArtificial SequenceSynthetic 253ggtgcatcc 92543PRTArtificial SequenceSynthetic 254Gly Ala Ser125530DNAArtificial SequenceSynthetic 255cagcagtata acaactggcc catgtacact 3025610PRTArtificial SequenceSynthetic 256Gln Gln Tyr Asn Asn Trp Pro Met Tyr Thr1 5 10257361DNAArtificial SequenceSynthetic 257caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgaag cctctggatt caccttcagt agttctggca tgcactgggt ccgccaggct 120ccaggcaagg ggctgcagtg ggtggctgtt atatcatatg atggaaataa taaattttat 180gaagactccg tgaagggccg attgaccatt tccagagaca attccaacaa cactctgtgg 240ctgcaaatga acagcctgag agttgaagac acggctgttt attactgtgc gaaatcagga 300ggtagagtgg gagccgcctt tgcctactgg ggccagggaa ccctggtcac cgtctcctca 360g 361258120PRTArtificial SequenceSynthetic 258Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Phe Thr Phe Ser Ser Ser 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Gln Trp Val 35 40 45Ala Val Ile Ser Tyr Asp Gly Asn Asn Lys Phe Tyr Glu Asp Ser Val 50 55 60Lys Gly Arg Leu Thr Ile Ser Arg Asp Asn Ser Asn Asn Thr Leu Trp65 70 75 80Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Lys Ser Gly Gly Arg Val Gly Ala Ala Phe Ala Tyr Trp Gly Gln 100 105 110Gly Thr Leu Val Thr Val Ser Ser 115 12025924DNAArtificial SequenceSynthetic 259ggattcacct tcagtagttc tggc 242608PRTArtificial SequenceSynthetic 260Gly Phe Thr Phe Ser Ser Ser Gly1 526124DNAArtificial SequenceSynthetic 261atatcatatg atggaaataa taaa 242628PRTArtificial SequenceSynthetic 262Ile Ser Tyr Asp Gly Asn Asn Lys1 526339DNAArtificial SequenceSynthetic 263gcgaaatcag gaggtagagt gggagccgcc tttgcctac 3926413PRTArtificial SequenceSynthetic 264Ala Lys Ser Gly Gly Arg Val Gly Ala Ala Phe Ala Tyr1 5 10265337DNAArtificial SequenceSynthetic 265gatattgtga acactcagtc tccactctct ctgcccgtca cccctggaga gccggcctcc 60atctcctgca ggtctagtca gagcctcctg tatggtaatg gatacaacta tttggattgg 120tacctgcaga agccagggca ctctccacag ctcctgatct atttgggttc taatcggggc 180tccggggtcc ctgacaggtt cagtggcagt ggatcaggca cagattttac actgaagatc 240agcagagtgg aggctgaaga tgttggcatt tattactgca tgcaaactct acaaactcca 300ttcactttcg gccctgggac caaaatgtat atcaaac 337266112PRTArtificial SequenceSynthetic 266Asp Ile Val Asn Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly1 5 10 15Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Tyr Gly 20 25 30Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly His Ser 35 40 45Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Gly Ser Gly Val Pro 50 55 60Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile65 70 75 80Ser Arg Val Glu Ala Glu Asp Val Gly Ile Tyr Tyr Cys Met Gln Thr 85 90 95Leu Gln Thr Pro Phe Thr Phe Gly Pro Gly Thr Lys Met Tyr Ile Lys 100 105 11026733DNAArtificial SequenceSynthetic 267cagagcctcc tgtatggtaa tggatacaac tat 3326811PRTArtificial SequenceSynthetic 268Gln Ser Leu Leu Tyr Gly Asn Gly Tyr Asn Tyr1 5 102699DNAArtificial SequenceSynthetic 269ttgggttct 92703PRTArtificial SequenceSynthetic 270Leu Gly Ser127127DNAArtificial SequenceSynthetic 271atgcaaactc tacaaactcc attcact 272729PRTArtificial SequenceSynthetic 272Met Gln Thr Leu Gln Thr Pro Phe Thr1 5273376DNAArtificial SequenceSynthetic 273gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag gctctggaat cacctttagc agctatgcca tgagctgggt ccgccaggct 120ccagggaagg ggctggagtg ggtctcagct attagtggta atggtggtac cacaaactac 180gcagactccg tgaagggccg gttcaccatc tccagagaca actccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaggac acggccgtat attactgtgc gaaagaaaga 300atccttacca gcagctggac gaggtacggt attatggacg tctggggcca agggaccacg 360gtcaccgtct cctcag 376274125PRTArtificial SequenceSynthetic 274Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Ile Thr Phe Ser Ser Tyr 20 25 30Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Ala Ile Ser Gly Asn Gly Gly Thr Thr Asn Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Lys Glu Arg Ile Leu Thr Ser Ser Trp Thr Arg Tyr Gly Ile Met 100 105 110Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 12527524DNAArtificial SequenceSynthetic 275ggaatcacct ttagcagcta tgcc 242768PRTArtificial SequenceSynthetic 276Gly Ile Thr Phe Ser Ser Tyr Ala1 527724DNAArtificial SequenceSynthetic 277attagtggta atggtggtac caca 242788PRTArtificial SequenceSynthetic 278Ile Ser Gly Asn Gly Gly Thr Thr1 527954DNAArtificial SequenceSynthetic 279gcgaaagaaa gaatccttac cagcagctgg acgaggtacg gtattatgga cgtc 5428018PRTArtificial SequenceSynthetic 280Ala Lys Glu Arg Ile Leu Thr Ser Ser Trp Thr Arg Tyr Gly Ile Met1 5 10 15Asp Val281322DNAArtificial SequenceSynthetic 281gaaatagtga tgacgcagtc tccagccacc ctgtctatgt ctccagggga aagagccacc 60ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacca gcagaaacct 120ggccaggctc ccaggctcct catctatggt gcatccacca gggccactgg tatcccagcc 180aggttcagtg gcagtgggtc tgggacagag ttcactctca ccatcagcag cctgcagtct 240gaagattttg cagtttatta ctgtcagcag tataataact ggcctctcac tttcggcgga 300gggaccaagt tagagatcaa ac 322282107PRTArtificial SequenceSynthetic 282Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Met Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Ser

Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Leu 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10528318DNAArtificial SequenceSynthetic 283cagagtgtta gcagcaac 182846PRTArtificial SequenceSynthetic 284Gln Ser Val Ser Ser Asn1 52859DNAArtificial SequenceSynthetic 285ggtgcatcc 92863PRTArtificial SequenceSynthetic 286Gly Ala Ser128727DNAArtificial SequenceSynthetic 287cagcagtata ataactggcc tctcact 272889PRTArtificial SequenceSynthetic 288Gln Gln Tyr Asn Asn Trp Pro Leu Thr1 5289358DNAArtificial SequenceSynthetic 289caggtgcaac tacagcagtg gggcgcagga ctgttgaagc cttcggagac cctgtccctc 60acctgcgctg tctatggtgg gtccttcagt ggttactact ggagctggat ccgccagccc 120ccagggaagg ggctggagtg gattggggaa atcaatcata gtggaaacac caactacaac 180ccgtccctca agagtcgagt caccatatca gtagacacgt ccaagaacca gttctccctg 240aaactgaact ctgtgaccgc cgcggacacg gctgtgtatt actgtgcgag aagagaggct 300acagtaactc catactttga ctactggggc cagggaaccc tggtcaccgt ctcctcag 358290119PRTArtificial SequenceSynthetic 290Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr 20 25 30Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Glu Ile Asn His Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys Leu Asn Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Arg Glu Ala Thr Val Thr Pro Tyr Phe Asp Tyr Trp Gly Gln Gly 100 105 110Thr Leu Val Thr Val Ser Ser 11529124DNAArtificial SequenceSynthetic 291ggtgggtcct tcagtggtta ctac 242928PRTArtificial SequenceSynthetic 292Gly Gly Ser Gly Ser Gly Tyr Tyr1 529321DNAArtificial SequenceSynthetic 293atcaatcata gtggaaacac c 212947PRTArtificial SequenceSynthetic 294Ile Asn His Ser Gly Asn Thr1 529539DNAArtificial SequenceSynthetic 295gcgagaagag aggctacagt aactccatac tttgactac 3929613PRTArtificial SequenceSynthetic 296Ala Arg Arg Glu Ala Thr Val Thr Pro Tyr Phe Asp Tyr1 5 10297322DNAArtificial SequenceSynthetic 297gacatccagt tgacccagtc tccatccttc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgct gggccagtca gggcattagc agttatttag cctggtatca gcaaaaacca 120gggaaagccc ctaagctcct gatctatgct gcatccactt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 240gaagattttg caacttatta ctgtcaacag cttaatagtt atccgctcac tttcggcgga 300gggaccaagg tggagatcaa ac 322298107PRTArtificial SequenceSynthetic 298Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Trp Ala Ser Gln Gly Ile Ser Ser Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Leu Asn Ser Tyr Pro Leu 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 10529918DNAArtificial SequenceSynthetic 299cagggcatta gcagttat 183006PRTArtificial SequenceSynthetic 300Gln Gly Ile Ser Ser Tyr1 53019DNAArtificial SequenceSynthetic 301gctgcatcc 93023PRTArtificial SequenceSynthetic 302Ala Ala Ser130327DNAArtificial SequenceSynthetic 303caacagctta atagttatcc gctcact 273049PRTArtificial SequenceSynthetic 304Gln Gln Leu Asn Ser Tyr Pro Leu Thr1 5305355DNAArtificial SequenceSynthetic 305caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgtag tctctggatt caacttcagt aggaatggca tacactgggt ccgccaggct 120ccaggcaagg ggctggaatg ggtggcagtt atatcatatg atggaagaaa taaattttat 180gtagagtccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagtctgag agttgaggac acggctgtat attactgtgc gaaatcctca 300attggagggt tttttgaata ctggggccag ggaaccctgg tcaccgtctc ctcag 355306118PRTArtificial SequenceSynthetic 306Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Val Val Ser Gly Phe Asn Phe Ser Arg Asn 20 25 30Gly Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser Tyr Asp Gly Arg Asn Lys Phe Tyr Val Glu Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Lys Ser Ser Ile Gly Gly Phe Phe Glu Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser 11530724DNAArtificial SequenceSynthetic 307ggattcaact tcagtaggaa tggc 243088PRTArtificial SequenceSynthetic 308Gly Phe Asn Phe Ser Arg Asn Gly1 530924DNAArtificial SequenceSynthetic 309atatcatatg atggaagaaa taaa 243108PRTArtificial SequenceSynthetic 310Ile Ser Tyr Asp Gly Arg Asn Lys1 531133DNAArtificial SequenceSynthetic 311gcgaaatcct caattggagg gttttttgaa tac 3331211PRTArtificial SequenceSynthetic 312Ala Lys Ser Ser Ile Gly Gly Phe Phe Glu Tyr1 5 10313337DNAArtificial SequenceSynthetic 313gatattgtga tgactcagtc tccactctcc ctgcccgtca ctcctggaga gccggcctcc 60atctcctgca ggtctagtca gagcctcctg catagtaatg gatacaacta tttggattgg 120tacctgcaga agccaggaca gtctccacaa ctcatgatct atttgggttc tcatcgggcc 180tccggggtcc ctgacaggtt cagtggcagt ggatcaggca cagattttac actgaaaatc 240agcagagtgg aggctgagga tgttggagtc tattactgca ttcaagttca acaaactccg 300atcaccttcg gccaagggac acggctggag attaaac 337314112PRTArtificial SequenceSynthetic 314Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly1 5 10 15Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser 20 25 30Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45Pro Gln Leu Met Ile Tyr Leu Gly Ser His Arg Ala Ser Gly Val Pro 50 55 60Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile65 70 75 80Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ile Gln Val 85 90 95Gln Gln Thr Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys 100 105 11031533DNAArtificial SequenceSynthetic 315cagagcctcc tgcatagtaa tggatacaac tat 3331611PRTArtificial SequenceSynthetic 316Gln Ser Leu Leu His Ser Asn Gly Tyr Asn Tyr1 5 103179DNAArtificial SequenceSynthetic 317ttgggttct 93183PRTArtificial SequenceSynthetic 318Leu Gly Ser131927DNAArtificial SequenceSynthetic 319attcaagttc aacaaactcc gatcacc 273209PRTArtificial SequenceSynthetic 320Ile Gln Val Gln Gln Thr Pro Ile Thr1 5321337DNAArtificial SequenceSynthetic 321gatattgtga tgactcagac tccactctcc tcacctgtca cccttggaca gccggcctcc 60atctcctgca ggtctagtca aagcctcgta cacagtgatg gaaacaccta cttgagttgg 120cttcagcaga ggccaggcca gcctccaaga ctcctaattt acaagatttc taaccggttc 180tctggggtcc cagacagatt cagtggcagt ggggcaggga cagatttcac actgaaaatc 240agcagggtgg aagctgagga tgtcggggtt tatttctgca tgcaagctac acaatttccg 300tacacttttg gccaggggac caagctggag atcaaag 337322112PRTArtificial SequenceSynthetic 322Asp Ile Val Met Thr Gln Thr Pro Leu Ser Ser Pro Val Thr Leu Gly1 5 10 15Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser 20 25 30Asp Gly Asn Thr Tyr Leu Ser Trp Leu Gln Gln Arg Pro Gly Gln Pro 35 40 45Pro Arg Leu Leu Ile Tyr Lys Ile Ser Asn Arg Phe Ser Gly Val Pro 50 55 60Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Lys Ile65 70 75 80Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Phe Cys Met Gln Ala 85 90 95Thr Gln Phe Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 11032333DNAArtificial SequenceSynthetic 323caaagcctcg tacacagtga tggaaacacc tac 3332411PRTArtificial SequenceSynthetic 324Gln Ser Leu Val His Ser Asp Gly Asn Thr Tyr1 5 103259DNAArtificial SequenceSynthetic 325aagatttct 93263PRTArtificial SequenceSynthetic 326Lys Ile Ser132727DNAArtificial SequenceSynthetic 327atgcaagcta cacaatttcc gtacact 273289PRTArtificial SequenceSynthetic 328Met Gln Ala Thr Gln Phe Pro Tyr Thr1 53298PRTArtificial SequenceSyntheticVARIANT(1)..(1)Xaa = GlyVARIANT(2)..(2)Xaa = PheVARIANT(3)..(3)Xaa = ThrVARIANT(4)..(4)Xaa = PheVARIANT(5)..(5)Xaa = SerVARIANT(6)..(6)Xaa = Ala or SerVARIANT(7)..(7)Xaa = Phe or TyrVARIANT(8)..(8)Xaa = Gly or Ala 329Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa1 53308PRTArtificial SequenceSyntheticVARIANT(1)..(1)Xaa = IleVARIANT(2)..(2)Xaa = Gly or SerVARIANT(3)..(3)Xaa = Tyr or GlyVARIANT(4)..(4)Xaa = Ser or AspVARIANT(5)..(5)Xaa = GlyVARIANT(6)..(6)Xaa = GlyVARIANT(7)..(7)Xaa = Ser or AsnVARIANT(8)..(8)Xaa = Ala or Glu 330Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa1 533114PRTArtificial SequenceSyntheticVARIANT(1)..(1)Xaa = Ser or AlaVARIANT(2)..(2)Xaa = Thr or LysVARIANT(3)..(3)Xaa = Asp or IleVARIANT(4)..(4)Xaa = Gly or SerVARIANT(5)..(5)Xaa = Ala or HisVARIANT(6)..(6)Xaa = Trp or TyrVARIANT(7)..(7)Xaa = Lys or AspVARIANT(8)..(8)Xaa = Met or IleVARIANT(9)..(9)Xaa = Ser or LeuVARIANT(10)..(10)Xaa = Gly or SerVARIANT(11)..(11)Xaa = Leu or GlyVARIANT(12)..(12)Xaa = Asp or MetVARIANT(13)..(13)Xaa = Val or AspVARIANT(14)..(14)Xaa = Val or absent 331Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa1 5 103326PRTArtificial SequenceSyntheticVARIANT(1)..(1)Xaa = GlnVARIANT(2)..(2)Xaa = Asp or GlyVARIANT(3)..(3)Xaa = IleVARIANT(4)..(4)Xaa = SerVARIANT(5)..(5)Xaa = Asp or AsnVARIANT(6)..(6)Xaa = Tyr or Trp 332Xaa Xaa Xaa Xaa Xaa Xaa1 53333PRTArtificial SequenceSyntheticVARIANT(1)..(1)Xaa = Thr or AlaVARIANT(2)..(2)Xaa = Thr or AlaVARIANT(3)..(3)Xaa = Ser 333Xaa Xaa Xaa13349PRTArtificial SequenceSyntheticVARIANT(1)..(1)Xaa = GlnVARIANT(2)..(2)Xaa = Lys or GlnVARIANT(3)..(3)Xaa = Ala or TyrVARIANT(4)..(4)Xaa = Asp or AsnVARIANT(5)..(5)Xaa = SerVARIANT(6)..(6)Xaa = Ala or PheVARIANT(7)..(7)Xaa = ProVARIANT(8)..(8)Xaa = LeuVARIANT(9)..(9)Xaa = Thr 334Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa1 5335330PRTArtificial SequenceSynthetic 335Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys1 5 10 15Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr65 70 75 80Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100 105 110Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 130 135 140Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp145 150 155 160Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210 215 220Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu225 230 235 240Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265 270Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275 280 285Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290 295 300Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr305 310 315 320Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330336327PRTArtificial SequenceSynthetic 336Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg1 5 10 15Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr65 70 75 80Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro 100 105 110Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 115 120 125Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 130 135 140Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp145 150 155 160Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe 165 170 175Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 180 185 190Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu 195 200 205Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 210 215 220Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys225 230 235 240Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 245 250 255Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 260 265 270Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 275 280 285Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser 290 295 300Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser305 310 315 320Leu Ser Leu Ser Leu Gly Lys 325337327PRTArtificial SequenceSynthetic 337Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg1 5 10 15Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35

40 45Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr65 70 75 80Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro 100 105 110Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 115 120 125Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 130 135 140Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp145 150 155 160Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe 165 170 175Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 180 185 190Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu 195 200 205Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 210 215 220Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys225 230 235 240Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 245 250 255Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 260 265 270Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 275 280 285Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser 290 295 300Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser305 310 315 320Leu Ser Leu Ser Leu Gly Lys 3253381128DNAHomo sapiens 338atgcaaaaac tgcaactctg tgtttatatt tacctgttta tgctgattgt tgctggtcca 60gtggatctaa atgagaacag tgagcaaaaa gaaaatgtgg aaaaagaggg gctgtgtaat 120gcatgtactt ggagacaaaa cactaaatct tcaagaatag aagccattaa gatacaaatc 180ctcagtaaac ttcgtctgga aacagctcct aacatcagca aagatgttat aagacaactt 240ttacccaaag ctcctccact ccgggaactg attgatcagt atgatgtcca gagggatgac 300agcagcgatg gctctttgga agatgacgat tatcacgcta caacggaaac aatcattacc 360atgcctacag agtctgattt tctaatgcaa gtggatggaa aacccaaatg ttgcttcttt 420aaatttagct ctaaaataca atacaataaa gtagtaaagg cccaactatg gatatatttg 480agacccgtcg agactcctac aacagtgttt gtgcaaatcc tgagactcat caaacctatg 540aaagacggta caaggtatac tggaatccga tctctgaaac ttgacatgaa cccaggcact 600ggtatttggc agagcattga tgtgaagaca gtgttgcaaa attggctcaa acaacctgaa 660tccaacttag gcattgaaat aaaagcttta gatgagaatg gtcatgatct tgctgtaacc 720ttcccaggac caggagaaga tgggctgaat ccgtttttag aggtcaaggt aacagacaca 780ccaaaaagat ccagaaggga ttttggtctt gactgtgatg agcactcaac agaatcacga 840tgctgtcgtt accctctaac tgtggatttt gaagcttttg gatgggattg gattatcgct 900cctaaaagat ataaggccaa ttactgctct ggagagtgtg aatttgtatt tttacaaaaa 960tatcctcata ctcatctggt acaccaagca aaccccagag gttcagcagg cccttgctgt 1020actcccacaa agatgtctcc aattaatatg ctatatttta atggcaaaga acaaataata 1080tatgggaaaa ttccagcgat ggtagtagac cgctgtgggt gctcatga 1128339375PRTHomo sapiens 339Met Gln Lys Leu Gln Leu Cys Val Tyr Ile Tyr Leu Phe Met Leu Ile1 5 10 15Val Ala Gly Pro Val Asp Leu Asn Glu Asn Ser Glu Gln Lys Glu Asn 20 25 30Val Glu Lys Glu Gly Leu Cys Asn Ala Cys Thr Trp Arg Gln Asn Thr 35 40 45Lys Ser Ser Arg Ile Glu Ala Ile Lys Ile Gln Ile Leu Ser Lys Leu 50 55 60Arg Leu Glu Thr Ala Pro Asn Ile Ser Lys Asp Val Ile Arg Gln Leu65 70 75 80Leu Pro Lys Ala Pro Pro Leu Arg Glu Leu Ile Asp Gln Tyr Asp Val 85 90 95Gln Arg Asp Asp Ser Ser Asp Gly Ser Leu Glu Asp Asp Asp Tyr His 100 105 110Ala Thr Thr Glu Thr Ile Ile Thr Met Pro Thr Glu Ser Asp Phe Leu 115 120 125Met Gln Val Asp Gly Lys Pro Lys Cys Cys Phe Phe Lys Phe Ser Ser 130 135 140Lys Ile Gln Tyr Asn Lys Val Val Lys Ala Gln Leu Trp Ile Tyr Leu145 150 155 160Arg Pro Val Glu Thr Pro Thr Thr Val Phe Val Gln Ile Leu Arg Leu 165 170 175Ile Lys Pro Met Lys Asp Gly Thr Arg Tyr Thr Gly Ile Arg Ser Leu 180 185 190Lys Leu Asp Met Asn Pro Gly Thr Gly Ile Trp Gln Ser Ile Asp Val 195 200 205Lys Thr Val Leu Gln Asn Trp Leu Lys Gln Pro Glu Ser Asn Leu Gly 210 215 220Ile Glu Ile Lys Ala Leu Asp Glu Asn Gly His Asp Leu Ala Val Thr225 230 235 240Phe Pro Gly Pro Gly Glu Asp Gly Leu Asn Pro Phe Leu Glu Val Lys 245 250 255Val Thr Asp Thr Pro Lys Arg Ser Arg Arg Asp Phe Gly Leu Asp Cys 260 265 270Asp Glu His Ser Thr Glu Ser Arg Cys Cys Arg Tyr Pro Leu Thr Val 275 280 285Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile Ile Ala Pro Lys Arg Tyr 290 295 300Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu Phe Val Phe Leu Gln Lys305 310 315 320Tyr Pro His Thr His Leu Val His Gln Ala Asn Pro Arg Gly Ser Ala 325 330 335Gly Pro Cys Cys Thr Pro Thr Lys Met Ser Pro Ile Asn Met Leu Tyr 340 345 350Phe Asn Gly Lys Glu Gln Ile Ile Tyr Gly Lys Ile Pro Ala Met Val 355 360 365Val Asp Arg Cys Gly Cys Ser 370 375340109PRTHomo sapiens 340Asp Phe Gly Leu Asp Cys Asp Glu His Ser Thr Glu Ser Arg Cys Cys1 5 10 15Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile 20 25 30Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu 35 40 45Phe Val Phe Leu Gln Lys Tyr Pro His Thr His Leu Val His Gln Ala 50 55 60Asn Pro Arg Gly Ser Ala Gly Pro Cys Cys Thr Pro Thr Lys Met Ser65 70 75 80Pro Ile Asn Met Leu Tyr Phe Asn Gly Lys Glu Gln Ile Ile Tyr Gly 85 90 95Lys Ile Pro Ala Met Val Val Asp Arg Cys Gly Cys Ser 100 1053411224DNAHomo sapiens 341atggtgctcg cggccccgct gctgctgggc ttcctgctcc tcgccctgga gctgcggccc 60cggggggagg cggccgaggg ccccgcggcg gcggcggcgg cggcggcggc ggcggcagcg 120gcgggggtcg ggggggagcg ctccagccgg ccagccccgt ccgtggcgcc cgagccggac 180ggctgccccg tgtgcgtttg gcggcagcac agccgcgagc tgcgcctaga gagcatcaag 240tcgcagatct tgagcaaact gcggctcaag gaggcgccca acatcagccg cgaggtggtg 300aagcagctgc tgcccaaggc gccgccgctg cagcagatcc tggacctaca cgacttccag 360ggcgacgcgc tgcagcccga ggacttcctg gaggaggacg agtaccacgc caccaccgag 420accgtcatta gcatggccca ggagacggac ccagcagtac agacagatgg cagccctctc 480tgctgccatt ttcacttcag ccccaaggtg atgttcacaa aggtactgaa ggcccagctg 540tgggtgtacc tacggcctgt accccgccca gccacagtct acctgcagat cttgcgacta 600aaacccctaa ctggggaagg gaccgcaggg ggagggggcg gaggccggcg tcacatccgt 660atccgctcac tgaagattga gctgcactca cgctcaggcc attggcagag catcgacttc 720aagcaagtgc tacacagctg gttccgccag ccacagagca actggggcat cgagatcaac 780gcctttgatc ccagtggcac agacctggct gtcacctccc tggggccggg agccgagggg 840ctgcatccat tcatggagct tcgagtccta gagaacacaa aacgttcccg gcggaacctg 900ggtctggact gcgacgagca ctcaagcgag tcccgctgct gccgatatcc cctcacagtg 960gactttgagg ctttcggctg ggactggatc atcgcaccta agcgctacaa ggccaactac 1020tgctccggcc agtgcgagta catgttcatg caaaaatatc cgcataccca tttggtgcag 1080caggccaatc caagaggctc tgctgggccc tgttgtaccc ccaccaagat gtccccaatc 1140aacatgctct acttcaatga caagcagcag attatctacg gcaagatccc tggcatggtg 1200gtggatcgct gtggctgctc ttaa 1224342407PRTHomo sapiens 342Met Val Leu Ala Ala Pro Leu Leu Leu Gly Phe Leu Leu Leu Ala Leu1 5 10 15Glu Leu Arg Pro Arg Gly Glu Ala Ala Glu Gly Pro Ala Ala Ala Ala 20 25 30Ala Ala Ala Ala Ala Ala Ala Ala Ala Gly Val Gly Gly Glu Arg Ser 35 40 45Ser Arg Pro Ala Pro Ser Val Ala Pro Glu Pro Asp Gly Cys Pro Val 50 55 60Cys Val Trp Arg Gln His Ser Arg Glu Leu Arg Leu Glu Ser Ile Lys65 70 75 80Ser Gln Ile Leu Ser Lys Leu Arg Leu Lys Glu Ala Pro Asn Ile Ser 85 90 95Arg Glu Val Val Lys Gln Leu Leu Pro Lys Ala Pro Pro Leu Gln Gln 100 105 110Ile Leu Asp Leu His Asp Phe Gln Gly Asp Ala Leu Gln Pro Glu Asp 115 120 125Phe Leu Glu Glu Asp Glu Tyr His Ala Thr Thr Glu Thr Val Ile Ser 130 135 140Met Ala Gln Glu Thr Asp Pro Ala Val Gln Thr Asp Gly Ser Pro Leu145 150 155 160Cys Cys His Phe His Phe Ser Pro Lys Val Met Phe Thr Lys Val Leu 165 170 175Lys Ala Gln Leu Trp Val Tyr Leu Arg Pro Val Pro Arg Pro Ala Thr 180 185 190Val Tyr Leu Gln Ile Leu Arg Leu Lys Pro Leu Thr Gly Glu Gly Thr 195 200 205Ala Gly Gly Gly Gly Gly Gly Arg Arg His Ile Arg Ile Arg Ser Leu 210 215 220Lys Ile Glu Leu His Ser Arg Ser Gly His Trp Gln Ser Ile Asp Phe225 230 235 240Lys Gln Val Leu His Ser Trp Phe Arg Gln Pro Gln Ser Asn Trp Gly 245 250 255Ile Glu Ile Asn Ala Phe Asp Pro Ser Gly Thr Asp Leu Ala Val Thr 260 265 270Ser Leu Gly Pro Gly Ala Glu Gly Leu His Pro Phe Met Glu Leu Arg 275 280 285Val Leu Glu Asn Thr Lys Arg Ser Arg Arg Asn Leu Gly Leu Asp Cys 290 295 300Asp Glu His Ser Ser Glu Ser Arg Cys Cys Arg Tyr Pro Leu Thr Val305 310 315 320Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile Ile Ala Pro Lys Arg Tyr 325 330 335Lys Ala Asn Tyr Cys Ser Gly Gln Cys Glu Tyr Met Phe Met Gln Lys 340 345 350Tyr Pro His Thr His Leu Val Gln Gln Ala Asn Pro Arg Gly Ser Ala 355 360 365Gly Pro Cys Cys Thr Pro Thr Lys Met Ser Pro Ile Asn Met Leu Tyr 370 375 380Phe Asn Asp Lys Gln Gln Ile Ile Tyr Gly Lys Ile Pro Gly Met Val385 390 395 400Val Asp Arg Cys Gly Cys Ser 405343109PRTHomo sapiens 343Asn Leu Gly Leu Asp Cys Asp Glu His Ser Ser Glu Ser Arg Cys Cys1 5 10 15Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile 20 25 30Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Gln Cys Glu 35 40 45Tyr Met Phe Met Gln Lys Tyr Pro His Thr His Leu Val Gln Gln Ala 50 55 60Asn Pro Arg Gly Ser Ala Gly Pro Cys Cys Thr Pro Thr Lys Met Ser65 70 75 80Pro Ile Asn Met Leu Tyr Phe Asn Asp Lys Gln Gln Ile Ile Tyr Gly 85 90 95Lys Ile Pro Gly Met Val Val Asp Arg Cys Gly Cys Ser 100 1053441503DNAHomo sapiens 344atgagactcc ccaaactcct cactttcttg ctttggtacc tggcttggct ggacctggaa 60ttcatctgca ctgtgttggg tgcccctgac ttgggccaga gaccccaggg gaccaggcca 120ggattggcca aagcagaggc caaggagagg ccccccctgg cccggaacgt cttcaggcca 180gggggtcaca gctatggtgg gggggccacc aatgccaatg ccagggcaaa gggaggcacc 240gggcagacag gaggcctgac acagcccaag aaggatgaac ccaaaaagct gccccccaga 300ccgggcggcc ctgaacccaa gccaggacac cctccccaaa caaggcaggc tacagcccgg 360actgtgaccc caaaaggaca gcttcccgga ggcaaggcac ccccaaaagc aggatctgtc 420cccagctcct tcctgctgaa gaaggccagg gagcccgggc ccccacgaga gcccaaggag 480ccgtttcgcc caccccccat cacaccccac gagtacatgc tctcgctgta caggacgctg 540tccgatgctg acagaaaggg aggcaacagc agcgtgaagt tggaggctgg cctggccaac 600accatcacca gctttattga caaagggcaa gatgaccgag gtcccgtggt caggaagcag 660aggtacgtgt ttgacattag tgccctggag aaggatgggc tgctgggggc cgagctgcgg 720atcttgcgga agaagccctc ggacacggcc aagccagcgg cccccggagg cgggcgggct 780gcccagctga agctgtccag ctgccccagc ggccggcagc cggcctcctt gctggatgtg 840cgctccgtgc caggcctgga cggatctggc tgggaggtgt tcgacatctg gaagctcttc 900cgaaacttta agaactcggc ccagctgtgc ctggagctgg aggcctggga acggggcagg 960gccgtggacc tccgtggcct gggcttcgac cgcgccgccc ggcaggtcca cgagaaggcc 1020ctgttcctgg tgtttggccg caccaagaaa cgggacctgt tctttaatga gattaaggcc 1080cgctctggcc aggacgataa gaccgtgtat gagtacctgt tcagccagcg gcgaaaacgg 1140cgggccccac tggccactcg ccagggcaag cgacccagca agaaccttaa ggctcgctgc 1200agtcggaagg cactgcatgt caacttcaag gacatgggct gggacgactg gatcatcgca 1260ccccttgagt acgaggcttt ccactgcgag gggctgtgcg agttcccatt gcgctcccac 1320ctggagccca cgaatcatgc agtcatccag accctgatga actccatgga ccccgagtcc 1380acaccaccca cctgctgtgt gcccacgcgg ctgagtccca tcagcatcct cttcattgac 1440tctgccaaca acgtggtgta taagcagtat gaggacatgg tcgtggagtc gtgtggctgc 1500agg 1503345501PRTHomo sapiens 345Met Arg Leu Pro Lys Leu Leu Thr Phe Leu Leu Trp Tyr Leu Ala Trp1 5 10 15Leu Asp Leu Glu Phe Ile Cys Thr Val Leu Gly Ala Pro Asp Leu Gly 20 25 30Gln Arg Pro Gln Gly Thr Arg Pro Gly Leu Ala Lys Ala Glu Ala Lys 35 40 45Glu Arg Pro Pro Leu Ala Arg Asn Val Phe Arg Pro Gly Gly His Ser 50 55 60Tyr Gly Gly Gly Ala Thr Asn Ala Asn Ala Arg Ala Lys Gly Gly Thr65 70 75 80Gly Gln Thr Gly Gly Leu Thr Gln Pro Lys Lys Asp Glu Pro Lys Lys 85 90 95Leu Pro Pro Arg Pro Gly Gly Pro Glu Pro Lys Pro Gly His Pro Pro 100 105 110Gln Thr Arg Gln Ala Thr Ala Arg Thr Val Thr Pro Lys Gly Gln Leu 115 120 125Pro Gly Gly Lys Ala Pro Pro Lys Ala Gly Ser Val Pro Ser Ser Phe 130 135 140Leu Leu Lys Lys Ala Arg Glu Pro Gly Pro Pro Arg Glu Pro Lys Glu145 150 155 160Pro Phe Arg Pro Pro Pro Ile Thr Pro His Glu Tyr Met Leu Ser Leu 165 170 175Tyr Arg Thr Leu Ser Asp Ala Asp Arg Lys Gly Gly Asn Ser Ser Val 180 185 190Lys Leu Glu Ala Gly Leu Ala Asn Thr Ile Thr Ser Phe Ile Asp Lys 195 200 205Gly Gln Asp Asp Arg Gly Pro Val Val Arg Lys Gln Arg Tyr Val Phe 210 215 220Asp Ile Ser Ala Leu Glu Lys Asp Gly Leu Leu Gly Ala Glu Leu Arg225 230 235 240Ile Leu Arg Lys Lys Pro Ser Asp Thr Ala Lys Pro Ala Ala Pro Gly 245 250 255Gly Gly Arg Ala Ala Gln Leu Lys Leu Ser Ser Cys Pro Ser Gly Arg 260 265 270Gln Pro Ala Ser Leu Leu Asp Val Arg Ser Val Pro Gly Leu Asp Gly 275 280 285Ser Gly Trp Glu Val Phe Asp Ile Trp Lys Leu Phe Arg Asn Phe Lys 290 295 300Asn Ser Ala Gln Leu Cys Leu Glu Leu Glu Ala Trp Glu Arg Gly Arg305 310 315 320Ala Val Asp Leu Arg Gly Leu Gly Phe Asp Arg Ala Ala Arg Gln Val 325 330 335His Glu Lys Ala Leu Phe Leu Val Phe Gly Arg Thr Lys Lys Arg Asp 340 345 350Leu Phe Phe Asn Glu Ile Lys Ala Arg Ser Gly Gln Asp Asp Lys Thr 355 360 365Val Tyr Glu Tyr Leu Phe Ser Gln Arg Arg Lys Arg Arg Ala Pro Leu 370 375 380Ala Thr Arg Gln Gly Lys Arg Pro Ser Lys Asn Leu Lys Ala Arg Cys385 390 395 400Ser Arg Lys Ala Leu His Val Asn Phe Lys Asp Met Gly Trp Asp Asp 405 410 415Trp Ile Ile Ala Pro Leu Glu Tyr Glu Ala Phe His Cys Glu Gly Leu 420 425 430Cys Glu Phe Pro Leu Arg Ser His Leu Glu Pro Thr Asn His Ala Val 435 440 445Ile Gln Thr Leu Met Asn Ser Met Asp Pro Glu Ser Thr Pro Pro Thr 450 455 460Cys Cys Val Pro Thr Arg Leu Ser Pro Ile Ser Ile Leu Phe Ile Asp465 470 475 480Ser Ala Asn Asn Val Val Tyr Lys Gln Tyr Glu Asp Met Val Val Glu 485 490 495Ser Cys Gly Cys Arg 500346120PRTHomo sapiens 346Ala Pro Leu Ala Thr Arg Gln Gly Lys Arg Pro Ser Lys Asn Leu Lys1 5 10

15Ala Arg Cys Ser Arg Lys Ala Leu His Val Asn Phe Lys Asp Met Gly 20 25 30Trp Asp Asp Trp Ile Ile Ala Pro Leu Glu Tyr Glu Ala Phe His Cys 35 40 45Glu Gly Leu Cys Glu Phe Pro Leu Arg Ser His Leu Glu Pro Thr Asn 50 55 60His Ala Val Ile Gln Thr Leu Met Asn Ser Met Asp Pro Glu Ser Thr65 70 75 80Pro Pro Thr Cys Cys Val Pro Thr Arg Leu Ser Pro Ile Ser Ile Leu 85 90 95Phe Ile Asp Ser Ala Asn Asn Val Val Tyr Lys Gln Tyr Glu Asp Met 100 105 110Val Val Glu Ser Cys Gly Cys Arg 115 120347107PRTArtificial SequenceSynthetic 347Gln Ala Lys His Lys Gln Arg Lys Arg Leu Lys Ser Ser Cys Arg Tyr1 5 10 15Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile Ile Ala 20 25 30Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu Phe Val 35 40 45Phe Leu Gln Lys Tyr Pro His Thr His Leu Val His Gln Ala Asn Pro 50 55 60Arg Gly Ser Ala Gly Pro Cys Cys Thr Pro Thr Lys Met Ser Pro Ile65 70 75 80Asn Met Leu Tyr Phe Asn Gly Lys Glu Gln Ile Ile Tyr Gly Lys Ile 85 90 95Pro Ala Met Val Val Asp Arg Cys Gly Cys Ser 100 105348109PRTArtificial SequenceSynthetic 348Ala Leu Asp Thr Asn Tyr Cys Phe Ser Ser Thr Glu Lys Asn Cys Cys1 5 10 15Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile 20 25 30Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu 35 40 45Phe Val Phe Leu Gln Lys Tyr Pro His Thr His Leu Val His Gln Ala 50 55 60Asn Pro Arg Gly Ser Ala Gly Pro Cys Cys Thr Pro Thr Lys Met Ser65 70 75 80Pro Ile Asn Met Leu Tyr Phe Asn Gly Lys Glu Gln Ile Ile Tyr Gly 85 90 95Lys Ile Pro Ala Met Val Val Asp Arg Cys Gly Cys Ser 100 105349111PRTArtificial SequenceSynthetic 349Asp Phe Gly Leu Asp Cys Asp Glu His Ser Thr Glu Ser Arg Cys Cys1 5 10 15Lys Arg His Pro Leu Tyr Val Asp Phe Ser Asp Val Gly Trp Asn Asp 20 25 30Trp Ile Val Ala Pro Pro Gly Tyr His Ala Phe Tyr Cys Ser Gly Glu 35 40 45Cys Glu Phe Val Phe Leu Gln Lys Tyr Pro His Thr His Leu Val His 50 55 60Gln Ala Asn Pro Arg Gly Ser Ala Gly Pro Cys Cys Thr Pro Thr Lys65 70 75 80Met Ser Pro Ile Asn Met Leu Tyr Phe Asn Gly Lys Glu Gln Ile Ile 85 90 95Tyr Gly Lys Ile Pro Ala Met Val Val Asp Arg Cys Gly Cys Ser 100 105 110350110PRTArtificial SequenceSynthetic 350Asp Phe Gly Leu Asp Cys Asp Glu His Ser Thr Glu Ser Arg Cys Cys1 5 10 15Val Arg Gln Leu Tyr Ile Asp Phe Arg Lys Asp Leu Gly Trp Lys Trp 20 25 30Ile His Glu Pro Lys Gly Tyr His Ala Asn Phe Cys Ser Gly Glu Cys 35 40 45Glu Phe Val Phe Leu Gln Lys Tyr Pro His Thr His Leu Val His Gln 50 55 60Ala Asn Pro Arg Gly Ser Ala Gly Pro Cys Cys Thr Pro Thr Lys Met65 70 75 80Ser Pro Ile Asn Met Leu Tyr Phe Asn Gly Lys Glu Gln Ile Ile Tyr 85 90 95Gly Lys Ile Pro Ala Met Val Val Asp Arg Cys Gly Cys Ser 100 105 110351114PRTArtificial SequenceSynthetic 351Asp Phe Gly Leu Asp Cys Asp Glu His Ser Thr Glu Ser Arg Cys Cys1 5 10 15Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile 20 25 30Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Pro 35 40 45Phe Pro Leu Ala Asp His Leu Asn Ser Thr Asn His Ala Ile Val Gln 50 55 60Thr Leu Val Asn Ser Val Asn Ser Lys Ile Pro Lys Ala Cys Cys Thr65 70 75 80Pro Thr Lys Met Ser Pro Ile Asn Met Leu Tyr Phe Asn Gly Lys Glu 85 90 95Gln Ile Ile Tyr Gly Lys Ile Pro Ala Met Val Val Asp Arg Cys Gly 100 105 110Cys Ser352112PRTArtificial SequenceSynthetic 352Asp Phe Gly Leu Asp Cys Asp Glu His Ser Thr Glu Ser Arg Cys Cys1 5 10 15Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile 20 25 30Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Pro 35 40 45Tyr Ile Trp Ser Leu Asp Thr Gln Tyr Ser Lys Val Leu Ala Leu Tyr 50 55 60Asn Gln His Asn Pro Gly Ala Ser Ala Ala Pro Cys Cys Thr Pro Thr65 70 75 80Lys Met Ser Pro Ile Asn Met Leu Tyr Phe Asn Gly Lys Glu Gln Ile 85 90 95Ile Tyr Gly Lys Ile Pro Ala Met Val Val Asp Arg Cys Gly Cys Ser 100 105 110353109PRTArtificial SequenceSynthetic 353Asp Phe Gly Leu Asp Cys Asp Glu His Ser Thr Glu Ser Arg Cys Cys1 5 10 15Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile 20 25 30Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu 35 40 45Phe Val Phe Leu Gln Lys Tyr Pro His Thr His Leu Val His Gln Ala 50 55 60Asn Ser Val Asn Ser Lys Ile Pro Cys Cys Thr Pro Thr Lys Met Ser65 70 75 80Pro Ile Asn Met Leu Tyr Phe Asn Gly Lys Glu Gln Ile Ile Tyr Gly 85 90 95Lys Ile Pro Ala Met Val Val Asp Arg Cys Gly Cys Ser 100 105354109PRTArtificial SequenceSynthetic 354Asp Phe Gly Leu Asp Cys Asp Glu His Ser Thr Glu Ser Arg Cys Cys1 5 10 15Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile 20 25 30Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu 35 40 45Phe Val Phe Leu Gln Lys Tyr Pro His Thr His Leu Val His Gln Ala 50 55 60Asn Pro Gly Ala Ser Ala Ala Pro Cys Cys Thr Pro Thr Lys Met Ser65 70 75 80Pro Ile Asn Met Leu Tyr Phe Asn Gly Lys Glu Gln Ile Ile Tyr Gly 85 90 95Lys Ile Pro Ala Met Val Val Asp Arg Cys Gly Cys Ser 100 105355109PRTArtificial SequenceSynthetic 355Asp Phe Gly Leu Asp Cys Asp Glu His Ser Thr Glu Ser Arg Cys Cys1 5 10 15Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile 20 25 30Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu 35 40 45Phe Val Phe Leu Gln Lys Tyr Pro His Thr His Leu Val His Gln Ala 50 55 60Asn Pro Arg Gly Ser Ala Gly Pro Cys Cys Val Pro Thr Glu Leu Ser65 70 75 80Ala Ile Ser Met Leu Tyr Leu Asp Glu Asn Glu Lys Val Val Leu Lys 85 90 95Asn Tyr Gln Asp Met Val Val Glu Gly Cys Gly Cys Ser 100 105356108PRTArtificial SequenceSynthetic 356Asp Phe Gly Leu Asp Cys Asp Glu His Ser Thr Glu Ser Arg Cys Cys1 5 10 15Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile 20 25 30Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu 35 40 45Phe Val Phe Leu Gln Lys Tyr Pro His Thr His Leu Val His Gln Ala 50 55 60Asn Pro Arg Gly Ser Ala Gly Pro Cys Cys Val Pro Gln Ala Leu Glu65 70 75 80Pro Leu Pro Ile Val Tyr Tyr Val Gly Arg Lys Pro Lys Val Glu Gln 85 90 95Leu Ser Asn Met Ile Val Arg Ser Cys Gly Cys Ser 100 105357109PRTArtificial SequenceSynthetic 357Asp Phe Gly Leu Asp Cys Asp Glu His Ser Thr Glu Ser Arg Cys Cys1 5 10 15Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile 20 25 30Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu 35 40 45Phe Val Phe Leu Gln Lys Tyr Pro His Thr His Leu Val His Gln Ala 50 55 60Asn Pro Arg Gly Ser Ala Gly Pro Cys Cys Thr Pro Thr Lys Met Ser65 70 75 80Pro Ile Asn Met Leu Tyr Phe Asn Gly Lys Glu Lys Val Val Leu Lys 85 90 95Asn Tyr Gln Asp Met Val Val Glu Gly Cys Gly Cys Ser 100 105358108PRTArtificial SequenceSynthetic 358Asp Phe Gly Leu Asp Cys Asp Glu His Ser Thr Glu Ser Arg Cys Cys1 5 10 15Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile 20 25 30Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu 35 40 45Phe Val Phe Leu Gln Lys Tyr Pro His Thr His Leu Val His Gln Ala 50 55 60Asn Pro Arg Gly Ser Ala Gly Pro Cys Cys Thr Pro Thr Lys Met Ser65 70 75 80Pro Ile Asn Met Leu Tyr Phe Asn Gly Lys Lys Pro Lys Val Glu Gln 85 90 95Leu Ser Asn Met Ile Val Arg Ser Cys Gly Cys Ser 100 105359360DNAArtificial SequenceSynthetic 359gaggtgcagg tgttggagtc tgggggagac ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag cctctggatt cacctttagt gcctatgcca tgacctgggt ccgccaggct 120ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cgcatactac 180gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacggtatat 240ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gaaagatggg 300gcctggaaaa tgtccggttt ggacgtctgg ggccaaggga ccacggtcat cgtctcctca 360360120PRTArtificial SequenceSynthetic 360Glu Val Gln Val Leu Glu Ser Gly Gly Asp Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ala Tyr 20 25 30Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Ala Ile Ser Gly Ser Gly Gly Ser Ala Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Lys Asp Gly Ala Trp Lys Met Ser Gly Leu Asp Val Trp Gly Gln 100 105 110Gly Thr Thr Val Ile Val Ser Ser 115 12036124DNAArtificial SequenceSynthetic 361ggattcacct ttagtgccta tgcc 243628PRTArtificial SequenceSynthetic 362Gly Phe Thr Phe Ser Ala Tyr Ala1 536324DNAArtificial SequenceSynthetic 363attagtggta gtggtggtag cgca 243648PRTArtificial SequenceSynthetic 364Ile Ser Gly Ser Gly Gly Ser Ala1 536539DNAArtificial SequenceSynthetic 365gcgaaagatg gggcctggaa aatgtccggt ttggacgtc 3936613PRTArtificial SequenceSynthetic 366Ala Lys Asp Gly Ala Trp Lys Met Ser Gly Leu Asp Val1 5 10367321DNAArtificial SequenceSynthetic 367gacatccaga tgacccagtc tccagcctcc ctgtctgcat ctgttggaga cagagtcacc 60atcacttgtc gggcgagtca ggacattagc gattatttag cctggtatca gcagaaacca 120gggaaaattc ctaggctcct gatctatact acatccactt tgcaatcagg ggtcccatct 180cggttccgtg gcagtgggtc tgggacagat ttcactctca ccatcagcag cctgcagcct 240gaagatgttg caacttatta ctgtcagaag tatgacagtg ccccgctcac tttcggcgga 300gggaccaagg tggagatcaa a 321368107PRTArtificial SequenceSynthetic 368Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Asp Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ile Pro Arg Leu Leu Ile 35 40 45Tyr Thr Thr Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Arg Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Val Ala Thr Tyr Tyr Cys Gln Lys Tyr Asp Ser Ala Pro Leu 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 10536918DNAArtificial SequenceSynthetic 369caggacatta gcgattat 183706PRTArtificial SequenceSynthetic 370Gln Asp Ile Ser Asp Tyr1 53719DNAArtificial SequenceSynthetic 371actacatcc 93723PRTArtificial SequenceSynthetic 372Thr Thr Ser137327DNAArtificial SequenceSynthetic 373cagaagtatg acagtgcccc gctcact 273749PRTArtificial SequenceSynthetic 374Gln Lys Tyr Asp Ser Ala Pro Leu Thr1 5375363DNAArtificial SequenceSynthetic 375caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaagtc cctgcgactc 60tcctgtgcag cgtctggatt caccttcagt agttttggca tgcattgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt attgggtatg atggaggtaa tgaatactat 180gccgactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgaat 240ctgcaaatga gcagcctgag agccgaagac acggctgtgt attattgttc gactataagt 300cattacgata ttttgagcgg tatggacgtc tggggccgag ggaccacggt caccgtctcc 360tca 363376121PRTArtificial SequenceSynthetic 376Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Lys1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Gly Tyr Asp Gly Gly Asn Glu Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Asn65 70 75 80Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ser Thr Ile Ser His Tyr Asp Ile Leu Ser Gly Met Asp Val Trp Gly 100 105 110Arg Gly Thr Thr Val Thr Val Ser Ser 115 12037724DNAArtificial SequenceSynthetic 377ggattcacct tcagtagttt tggc 243788PRTArtificial SequenceSynthetic 378Gly Phe Thr Phe Ser Ser Phe Gly1 537924DNAArtificial SequenceSynthetic 379attgggtatg atggaggtaa tgaa 243808PRTArtificial SequenceSynthetic 380Ile Gly Tyr Asp Gly Gly Asn Glu1 538142DNAArtificial SequenceSynthetic 381tcgactataa gtcattacga tattttgagc ggtatggacg tc 4238214PRTArtificial SequenceSynthetic 382Ser Thr Ile Ser His Tyr Asp Ile Leu Ser Gly Met Asp Val1 5 10383324DNAArtificial SequenceSynthetic 383gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtaggaga cagagtcacc 60atcacttgtc gggcgagtca gggtattagc aactggttag cctggtatca gcagaaacca 120gggaaagccc ctaagctcct gatctttgct gcatccagtt tgcaaagtgg ggtcccatca 180cggttcagcg gcagtgcatc tgggacagat ttcactctca ccatcaacag cctgcagcct 240gaagattttg caacttacta ttgtcaacag gctaacagtt tcccgctcac tttcggcgga 300gggaccaagg tggagatcaa acga 324384108PRTArtificial SequenceSynthetic 384Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Asn Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Phe Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Ala Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn Ser

Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Leu 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg 100 10538518DNAArtificial SequenceSynthetic 385cagggtatta gcaactgg 183866PRTArtificial SequenceSynthetic 386Gln Gly Ile Ser Asn Trp1 53879DNAArtificial SequenceSynthetic 387gctgcatcc 93883PRTArtificial SequenceSynthetic 388Ala Ala Ser138927DNAArtificial SequenceSynthetic 389caacaggcta acagtttccc gctcact 273909PRTArtificial SequenceSynthetic 390Gln Gln Ala Asn Ser Phe Pro Leu Thr1 5391340PRTArtificial SequenceSynthetic 391Glu Ala Glu Thr Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu1 5 10 15Glu Arg Thr Asn Gln Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp 20 25 30Lys Arg Leu His Cys Tyr Ala Ser Trp Ala Asn Ser Ser Gly Thr Ile 35 40 45Glu Leu Val Lys Lys Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp 50 55 60Arg Gln Glu Cys Val Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys65 70 75 80Cys Cys Glu Gly Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu 85 90 95Ala Gly Gly Pro Glu Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Ser 100 105 110Gly Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 115 120 125Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 130 135 140Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser145 150 155 160His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 165 170 175Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 180 185 190Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 195 200 205Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 210 215 220Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln225 230 235 240Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 245 250 255Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 260 265 270Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 275 280 285Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 290 295 300Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val305 310 315 320Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 325 330 335Ser Pro Gly Lys 340392366DNAArtificial SequenceSynthetic 392caggtacagc tgcagcagtc aggtccagga ctgctgaagc cctcgcagac cctctcactc 60acctgtgcca tctccgggga cagtgtctct agcaacagtg ctgcttggag ttggatcagg 120cagtccccat cgagaggcct tgagtggctg ggaaggacat attacagggc caactggttt 180aatgattatg cactttctgt gaaaagtcga ataaccatca acccagtcac atccacgaac 240cacttctccc tgcagctgca ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300agagaagggg ctctgggata ctactttgac tcctggggcc agggaaccct ggtcaccgtc 360tcctca 366393122PRTArtificial SequenceSynthetic 393Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Leu Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Ser Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ala Asn Trp Phe Asn Asp Tyr Ala 50 55 60Leu Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Val Thr Ser Thr Asn65 70 75 80His Phe Ser Leu Gln Leu His Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Glu Gly Ala Leu Gly Tyr Tyr Phe Asp Ser Trp 100 105 110Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 12039430DNAArtificial SequenceSynthetic 394ggggacagtg tctctagcaa cagtgctgct 3039510PRTArtificial SequenceSynthetic 395Gly Asp Ser Val Ser Ser Asn Ser Ala Ala1 5 1039627DNAArtificial SequenceSynthetic 396acatattaca gggccaactg gtttaat 273979PRTArtificial SequenceSynthetic 397Thr Tyr Tyr Arg Ala Asn Trp Phe Asn1 539836DNAArtificial SequenceSynthetic 398gcaagagaag gggctctggg atactacttt gactcc 3639912PRTArtificial SequenceSynthetic 399Ala Arg Glu Gly Ala Leu Gly Tyr Tyr Phe Asp Ser1 5 10400339DNAArtificial SequenceSynthetic 400gacatcgtga tgacccagtc tccagactcc ctggctgtgt ctctgggcga gagggccacc 60atcaactgca agtccagtca aagtgtttta tacagctcca acaataagaa ttatttagct 120tggtaccaac agaaaccagg gcagcctcct acactgctca tttactgggc atctacccgg 180gaatccgggg tccctgaccg attcagtggc agcgggtctg ggacagattt cactctcacc 240atcagcagcc tgcaggcgga agatgtggca atttattact gtcaccaata ttttattact 300ccactcactt tcggcggagg gaccaaggtg gagatcaaa 339401113PRTArtificial SequenceSynthetic 401Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly1 5 10 15Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser 20 25 30Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45Pro Pro Thr Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr65 70 75 80Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Ile Tyr Tyr Cys His Gln 85 90 95Tyr Phe Ile Thr Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 100 105 110Lys40236DNAArtificial SequenceSynthetic 402caaagtgttt tatacagctc caacaataag aattat 3640312PRTArtificial SequenceSynthetic 403Gln Ser Val Leu Tyr Ser Ser Asn Asn Lys Asn Tyr1 5 104049DNAArtificial SequenceSynthetic 404tgggcatct 94053PRTArtificial SequenceSynthetic 405Trp Ala Ser140627DNAArtificial SequenceSynthetic 406caccaatatt ttattactcc actcact 274079PRTArtificial SequenceSynthetic 407His Gln Tyr Phe Ile Thr Pro Leu Thr1 5408378DNAArtificial SequenceSynthetic 408caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt atatggtatg atggaagtaa taaatacaat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacggtgtat 240ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc gagagcccgg 300aattacgata ttttgactgg ttattataac ctcggtatgg acgtctgggg ccaagggacc 360acggtcaccg tctcctca 378409126PRTArtificial SequenceSynthetic 409Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Asp Gly Ser Asn Lys Tyr Asn Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Ala Arg Asn Tyr Asp Ile Leu Thr Gly Tyr Tyr Asn Leu Gly 100 105 110Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 12541024DNAArtificial SequenceSynthetic 410ggattcacct tcagtagcta tggc 244118PRTArtificial SequenceSynthetic 411Gly Phe Thr Phe Ser Ser Tyr Gly1 541224DNAArtificial SequenceSynthetic 412atatggtatg atggaagtaa taaa 244138PRTArtificial SequenceSynthetic 413Ile Trp Tyr Asp Gly Ser Asn Lys1 541457DNAArtificial SequenceSynthetic 414gcgagagccc ggaattacga tattttgact ggttattata acctcggtat ggacgtc 5741523PRTArtificial SequenceSynthetic 415Ala Arg Ala Arg Asn Tyr Asp Ile Leu Thr Gly Tyr Tyr Asn Leu Gly1 5 10 15Met Asp Val Ser Pro Gly Lys 20416321DNAArtificial SequenceSynthetic 416gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca gggcattaga aatgatttag gctggtatca acagaaacca 120gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 240gaagattttg caacttatta ctgtctacag cataatagtt acccgtacac ttttggccag 300gggaccaagc tggagatcaa a 321417107PRTArtificial SequenceSynthetic 417Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Asn Asp 20 25 30Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 10541818DNAArtificial SequenceSynthetic 418cagggcatta gaaatgat 184196PRTArtificial SequenceSynthetic 419Gln Gly Ile Arg Asn Asp1 54209DNAArtificial SequenceSynthetic 420gctgcatcc 94213PRTArtificial SequenceSynthetic 421Ala Ala Ser142227DNAArtificial SequenceSynthetic 422ctacagcata atagttaccc gtacact 274239PRTArtificial SequenceSynthetic 423Leu Gln His Asn Ser Tyr Pro Tyr Thr1 5424375DNAArtificial SequenceSynthetic 424gaagtgcagc tggtggagtc tgggggaaac ttggtacagt ctggcaggtc cctgagactc 60tcctgtacag cctctggatt cgcctttgat gattttgcca tgcactgggt ccggcaagtt 120ccagggaagg gcctggagtg ggtctcaggt attagttgga atagtgatac catcggctat 180gcggactctg tgaagggccg attcaccatt tccagagaca acgcccagaa ctccctgttt 240ctgcaaatgg acagtctgag agctgaggac acggccttgt attactgtgt aaaagatatg 300gttcggggac ttataggcta ctactactac ggtatggacg tctggggcca agggaccacg 360gtcaccgtct cctca 375425125PRTArtificial SequenceSynthetic 425Glu Val Gln Leu Val Glu Ser Gly Gly Asn Leu Val Gln Ser Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Ala Phe Asp Asp Phe 20 25 30Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Gly Ile Ser Trp Asn Ser Asp Thr Ile Gly Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Gln Asn Ser Leu Phe65 70 75 80Leu Gln Met Asp Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95Val Lys Asp Met Val Arg Gly Leu Ile Gly Tyr Tyr Tyr Tyr Gly Met 100 105 110Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 12542624DNAArtificial SequenceSynthetic 426ggattcgcct ttgatgattt tgcc 244278PRTArtificial SequenceSynthetic 427Gly Phe Ala Phe Asp Asp Phe Ala1 542824DNAArtificial SequenceSynthetic 428attagttgga atagtgatac catc 244298PRTArtificial SequenceSynthetic 429Ile Ser Trp Asn Ser Asp Thr Ile1 543054DNAArtificial SequenceSynthetic 430gtaaaagata tggttcgggg acttataggc tactactact acggtatgga cgtc 5443118PRTArtificial SequenceSynthetic 431Val Lys Asp Met Val Arg Gly Leu Ile Gly Tyr Tyr Tyr Tyr Gly Met1 5 10 15Asp Val432321DNAArtificial SequenceSynthetic 432gaaatagtgt tgacgcagtc tccagccatc ctgtctttgt ctccagggga aagagccatc 60ctctcctgca gggccagtca gagtatttac acctacttat cctggtacca acagacacct 120ggccgggctc ccaggctcct catctatgag acatccagca gggccactgg catcccagcc 180aggttcattg gcagtgggtc tgggacagac ttcactctca ccatcagtag cctagagcct 240gaagattttg cattttatta ctgtcagcac cgtagcgact ggcctcccac ttttggccag 300gggaccaagc tggagatcaa a 321433107PRTArtificial SequenceSynthetic 433Glu Ile Val Leu Thr Gln Ser Pro Ala Ile Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Ile Leu Ser Cys Arg Ala Ser Gln Ser Ile Tyr Thr Tyr 20 25 30Leu Ser Trp Tyr Gln Gln Thr Pro Gly Arg Ala Pro Arg Leu Leu Ile 35 40 45Tyr Glu Thr Ser Ser Arg Ala Thr Gly Ile Pro Ala Arg Phe Ile Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro65 70 75 80Glu Asp Phe Ala Phe Tyr Tyr Cys Gln His Arg Ser Asp Trp Pro Pro 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 10543418DNAArtificial SequenceSynthetic 434cagagtattt acacctac 184356PRTArtificial SequenceSynthetic 435Gln Ser Ile Tyr Thr Tyr1 54369DNAArtificial SequenceSynthetic 436gagacatcc 94373PRTArtificial SequenceSynthetic 437Glu Thr Ser143827DNAArtificial SequenceSynthetic 438cagcaccgta gcgactggcc tcccact 274399PRTArtificial SequenceSynthetic 439Gln His Arg Ser Asp Trp Pro Pro Thr1 5440351DNAArtificial SequenceSynthetic 440caggtgcagc tggtggagtc ggggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgtag cgtctggatt caccgtcagt agttatggca ttcactgggt ccgccaggct 120ccaggcaagg gactggagtg ggtgtcagtt atatggtatg atggaagaaa taaagactat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacggtgtat 240ttggaaatga aaggcctgag agccgaggac acggctcttt attattgtgc gagagacaaa 300actggggatt ttgactcctg gggccaggga accctggtca ccgtctcctc a 351441117PRTArtificial SequenceSynthetic 441Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Thr Val Ser Ser Tyr 20 25 30Gly Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Val Ile Trp Tyr Asp Gly Arg Asn Lys Asp Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr65 70 75 80Leu Glu Met Lys Gly Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95Ala Arg Asp Lys Thr Gly Asp Phe Asp Ser Trp Gly Gln Gly Thr Leu 100 105 110Val Thr Val Ser Ser 11544224DNAArtificial SequenceSynthetic 442ggattcaccg tcagtagtta tggc 244438PRTArtificial SequenceSynthetic 443Gly Phe Thr Val Ser Ser Tyr Gly1 544424DNAArtificial SequenceSynthetic 444atatggtatg atggaagaaa taaa 244458PRTArtificial SequenceSynthetic 445Ile Trp Tyr Asp Gly Arg Asn Lys1 544630DNAArtificial SequenceSynthetic 446gcgagagaca aaactgggga ttttgactcc 3044710PRTArtificial SequenceSynthetic 447Ala Arg Asp Lys Thr Gly Asp Phe Asp Ser1 5 10448321DNAArtificial SequenceSynthetic 448gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggtga cagagtcacc 60atcacttgcc gggcaagtca gaacattaac agctttttaa gttggtatca gcagaaacca 120ggaaaagccc ctaagttcct gatctatgat gcttccagta

tacaaagtgg ggccccatcg 180aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagtag tctgcaacct 240gaagattttg caacttacta ctgtcaacag agttacagtt ccccgttcac ttttggccag 300gggaccaagc tggagatcaa a 321449111PRTArtificial SequenceSynthetic 449Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Asn Ser Phe 20 25 30Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile 35 40 45Tyr Asp Ala Ser Ser Ile Gln Ser Gly Ala Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Ser Pro Phe 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Ser Pro Gly Lys 100 105 11045018DNAArtificial SequenceSynthetic 450cagaacatta acagcttt 184516PRTArtificial SequenceSynthetic 451Gln Asn Ile Asn Ser Phe1 54529DNAArtificial SequenceSynthetic 452gatgcttcc 94533PRTArtificial SequenceSynthetic 453Asp Ala Ser145427DNAArtificial SequenceSynthetic 454caacagagtt acagttcccc gttcact 274559PRTArtificial SequenceSynthetic 455Gln Gln Ser Tyr Ser Ser Pro Phe Thr1 5456375DNAArtificial SequenceSynthetic 456gaagtgcagc tggtggagtc tgggggaggc ttggtacagc ctggcaggtc cctgagactc 60tcctgtaaag cctctggatt cgcctttgat gatttcgcca tgcactgggt ccggcaagct 120ccagggaagg gcctggagtg ggtctcaggt attgtttgga acagtggtga cataggctat 180gcggactctg tgaagggccg attcaccatc tccagagaca acgccaagaa ctccctgtat 240ctgcaactga atagtctgag aactgaggac acggccttgt atttctgtgt aaaagatatg 300gttcggggac ttatgggctt caactattac ggtatggacg tctggggcca agggaccacg 360gtcaccgtct cctca 375457125PRTArtificial SequenceSynthetic 457Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Lys Ala Ser Gly Phe Ala Phe Asp Asp Phe 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Gly Ile Val Trp Asn Ser Gly Asp Ile Gly Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65 70 75 80Leu Gln Leu Asn Ser Leu Arg Thr Glu Asp Thr Ala Leu Tyr Phe Cys 85 90 95Val Lys Asp Met Val Arg Gly Leu Met Gly Phe Asn Tyr Tyr Gly Met 100 105 110Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 12545824DNAArtificial SequenceSynthetic 458ggattcgcct ttgatgattt cgcc 244598PRTArtificial SequenceSynthetic 459Gly Phe Ala Phe Asp Asp Phe Ala1 546024DNAArtificial SequenceSynthetic 460attgtttgga acagtggtga cata 244618PRTArtificial SequenceSynthetic 461Ile Val Trp Asn Ser Gly Asp Ile1 546254DNAArtificial SequenceSynthetic 462gtaaaagata tggttcgggg acttatgggc ttcaactatt acggtatgga cgtc 5446318PRTArtificial SequenceSynthetic 463Val Lys Asp Met Val Arg Gly Leu Met Gly Phe Asn Tyr Tyr Gly Met1 5 10 15Asp Val464321DNAArtificial SequenceSynthetic 464gaaattgtgt tgacgcagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60ctctcctgca gggccagtca aactattagt acttatttag tctggtaccg acagagacct 120ggccaggctc ccagtctcct catttatgat gcatccaaca gggccactga catcccagcc 180aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag ccttgagcct 240gaagattttg cagtttatta ctgtcagcag cgtagcaact ggccgatcac cttcggccaa 300gggacacgac tggagattaa a 321465107PRTArtificial SequenceSynthetic 465Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Thr Ile Ser Thr Tyr 20 25 30Leu Val Trp Tyr Arg Gln Arg Pro Gly Gln Ala Pro Ser Leu Leu Ile 35 40 45Tyr Asp Ala Ser Asn Arg Ala Thr Asp Ile Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Ile 85 90 95Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys 100 10546618DNAArtificial SequenceSynthetic 466caaactatta gtacttat 184676PRTArtificial SequenceSynthetic 467Gln Thr Ile Ser Thr Tyr1 54689DNAArtificial SequenceSynthetic 468gatgcatcc 94693PRTArtificial SequenceSynthetic 469Asp Ala Ser147027DNAArtificial SequenceSynthetic 470cagcagcgta gcaactggcc gatcacc 274719PRTArtificial SequenceSynthetic 471Gln Gln Arg Ser Asn Trp Pro Ile Thr1 5472369DNAArtificial SequenceSynthetic 472gaagtgcagc tggtggagtc tgggggaggc ttggtacagc ctggcaggtc cctgacactc 60tcctgtgcag tctctggatt cacctttgat gatcatgcca tgcactgggt ccggcaagct 120ccagggaagg gcctggagtg ggtctcaggt attagttgga atagtgtaag tataggctat 180gcggactctg tgaagggccg attcacgatc tccagagaca acgccaagac ctccctctat 240ctgcaaatga acagtctgag agttgacgac acggccttat attactgtgt gcaagattta 300aacgatattt tgactggtta tcccctcttt gacttttggg gccagggaac cctggtcacc 360gtctcctca 369473123PRTArtificial SequenceSynthetic 473Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg1 5 10 15Ser Leu Thr Leu Ser Cys Ala Val Ser Gly Phe Thr Phe Asp Asp His 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Gly Ile Ser Trp Asn Ser Val Ser Ile Gly Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Ser Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Val Asp Asp Thr Ala Leu Tyr Tyr Cys 85 90 95Val Gln Asp Leu Asn Asp Ile Leu Thr Gly Tyr Pro Leu Phe Asp Phe 100 105 110Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 12047424DNAArtificial SequenceSynthetic 474ggattcacct ttgatgatca tgcc 244758PRTArtificial SequenceSynthetic 475Gly Phe Thr Phe Asp Asp His Ala1 547624DNAArtificial SequenceSynthetic 476attagttgga atagtgtaag tata 244778PRTArtificial SequenceSynthetic 477Ile Ser Trp Asn Ser Val Ser Ile1 547848DNAArtificial SequenceSynthetic 478gtgcaagatt taaacgatat tttgactggt tatcccctct ttgacttt 4847916PRTArtificial SequenceSynthetic 479Val Gln Asp Leu Asn Asp Ile Leu Thr Gly Tyr Pro Leu Phe Asp Phe1 5 10 15480324DNAArtificial SequenceSynthetic 480gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca gagcattagc agctatttaa attggtatca gcagaaacca 120gggaaagccc ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccgtca 180aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct 240gaagattttg caacttacta ctgtcaacag agttacagta cccctccgat caccttcggc 300caagggacac gactggagat taaa 324481108PRTArtificial SequenceSynthetic 481Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr 20 25 30Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Pro 85 90 95Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys 100 10548218DNAArtificial SequenceSynthetic 482cagagcatta gcagctat 1848310PRTArtificial SequenceSynthetic 483Gln Ser Ile Ser Ser Tyr Ser Pro Gly Lys1 5 104849DNAArtificial SequenceSynthetic 484gctgcatcc 94853PRTArtificial SequenceSynthetic 485Ala Ala Ser148630DNAArtificial SequenceSynthetic 486caacagagtt acagtacccc tccgatcacc 3048710PRTArtificial SequenceSynthetic 487Gln Gln Ser Tyr Ser Thr Pro Pro Ile Thr1 5 10488369DNAArtificial SequenceSynthetic 488gaagtgcagc tggtggagtc tgggggaggc ttggtacagg ctggcaggtc cctaagactc 60tcctgtgaag cctctggatt cacctttgat gattatggca tgcactgggt ccggcaaggt 120ccagggaagg gcctggagtg ggtctcaggt attagttgga atagtggtaa catagactat 180gcggactctg tgaagggccg attcaccatc tccagagaca acgccaagac ctccctgtat 240ctgcaaatga acagtctgaa aactgacgac acggccttgt atttctgtgc aaaagatgct 300gtggggttta actggaacta ctttctcttt gactactggg gccagggaac cctggtcacc 360gtctcctca 369489123PRTArtificial SequenceSynthetic 489Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30Gly Met His Trp Val Arg Gln Gly Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Gly Ile Ser Trp Asn Ser Gly Asn Ile Asp Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Ser Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Lys Thr Asp Asp Thr Ala Leu Tyr Phe Cys 85 90 95Ala Lys Asp Ala Val Gly Phe Asn Trp Asn Tyr Phe Leu Phe Asp Tyr 100 105 110Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 12049024DNAArtificial SequenceSynthetic 490ggattcacct ttgatgatta tggc 244918PRTArtificial SequenceSynthetic 491Gly Phe Thr Phe Asp Asp Tyr Gly1 549224DNAArtificial SequenceSynthetic 492attagttgga atagtggtaa cata 244938PRTArtificial SequenceSynthetic 493Ile Ser Trp Asn Ser Gly Asn Ile1 549448DNAArtificial SequenceSynthetic 494gcaaaagatg ctgtggggtt taactggaac tactttctct ttgactac 4849516PRTArtificial SequenceSynthetic 495Ala Lys Asp Ala Val Gly Phe Asn Trp Asn Tyr Phe Leu Phe Asp Tyr1 5 10 15496351DNAArtificial SequenceSynthetic 496caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60acctgtgtag cgtctggatt caccgtcagt agttatggaa tgcactgggt ccgccaggcc 120ccaggcaagg ggctggagtg ggtggcagtt atgttttatg atgaaagtaa aaaatattat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agtcgaggac acggctgtgt attactgtgc gagagatgaa 300cagctcgact ttgaatactg gggccaggga accctggtca ccgtctcctc a 351497117PRTArtificial SequenceSynthetic 497Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Thr Cys Val Ala Ser Gly Phe Thr Val Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Met Phe Tyr Asp Glu Ser Lys Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Asp Glu Gln Leu Asp Phe Glu Tyr Trp Gly Gln Gly Thr Leu 100 105 110Val Thr Val Ser Ser 11549824DNAArtificial SequenceSynthetic 498ggattcaccg tcagtagtta tgga 244998PRTArtificial SequenceSynthetic 499Gly Phe Thr Val Ser Ser Tyr Gly1 550024DNAArtificial SequenceSynthetic 500atgttttatg atgaaagtaa aaaa 245018PRTArtificial SequenceSynthetic 501Met Phe Tyr Asp Glu Ser Lys Lys1 550230DNAArtificial SequenceSynthetic 502gcgagagatg aacagctcga ctttgaatac 3050310PRTArtificial SequenceSynthetic 503Ala Arg Asp Glu Gln Leu Asp Phe Glu Tyr1 5 10504369DNAArtificial SequenceSynthetic 504gaagtgcagc tggtggagtc tgggggaggc ttggtacagc ctggcaggtc cctgagactc 60tcctgtgcag cctctggatt cacctttgat gattatgcca tgcactgggt ccggcaagct 120ccagggaagg gcctggagtg ggtctcaggt attagttgga atagtggtag cataggctat 180gcggactctg tgaagggccg attcaccatc tccagagaca acgccaagaa ctccctgtat 240ctgcaaatga acagtctgag agctgaggac acggccttgt attactgtgc aaaagatata 300atggggaact gggactactt ctacggtatg gacgtctggg gccaagggac cacggtcacc 360gtctcctca 369505123PRTArtificial SequenceSynthetic 505Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95Ala Lys Asp Ile Met Gly Asn Trp Asp Tyr Phe Tyr Gly Met Asp Val 100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 12050624DNAArtificial SequenceSynthetic 506ggattcacct ttgatgatta tgcc 245078PRTArtificial SequenceSynthetic 507Gly Phe Thr Phe Asp Asp Tyr Ala1 550824DNAArtificial SequenceSynthetic 508attagttgga atagtggtag cata 245098PRTArtificial SequenceSynthetic 509Ile Ser Trp Asn Ser Gly Ser Ile1 551048DNAArtificial SequenceSynthetic 510gcaaaagata taatggggaa ctgggactac ttctacggta tggacgtc 4851116PRTArtificial SequenceSynthetic 511Ala Lys Asp Ile Met Gly Asn Trp Asp Tyr Phe Tyr Gly Met Asp Val1 5 10 15512369DNAArtificial SequenceSynthetic 512gaagtgcagc tggtggagtc tgggggaggc ttggtacagc ctggcaggtc cctgagactc 60tcctgtgcag cctctggatt cacctttgat gataatgcca tgcactgggt ccggcaacct 120ccagggaagg gcctggagtg ggtctcaggt attagttgga atagtggaag cataggctat 180gcggactctg tgaagggccg attcaccatc tccagagaca acgccaagaa ctccctgtat 240ctgcaaatga acagtctgag agctgaggac acggccttgt attactgtgc aaaagatata 300aacgatattt tgactggtta tcctcttttt gattactggg gccagggaac cctggtcacc 360gtctcctca 369513123PRTArtificial SequenceSynthetic 513Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Asn 20 25 30Ala Met His Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95Ala Lys Asp Ile Asn Asp Ile Leu Thr Gly Tyr Pro Leu Phe Asp Tyr 100 105 110Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 12051424DNAArtificial SequenceSynthetic 514ggattcacct ttgatgataa tgcc 245158PRTArtificial SequenceSynthetic 515Gly Phe Thr Phe Asp Asp Asn Ala1

551624DNAArtificial SequenceSynthetic 516attagttgga atagtggaag cata 2451712PRTArtificial SequenceSynthetic 517Ile Ser Trp Asn Ser Gly Ser Ile Ser Pro Gly Lys1 5 1051848DNAArtificial SequenceSynthetic 518gcaaaagata taaacgatat tttgactggt tatcctcttt ttgattac 4851916PRTArtificial SequenceSynthetic 519Ala Lys Asp Ile Asn Asp Ile Leu Thr Gly Tyr Pro Leu Phe Asp Tyr1 5 10 15520378DNAArtificial SequenceSynthetic 520gaagtgcagc tggtggagtc tgggggaggc gtggtacagc ctggggggtc cctgagactc 60tcctgtgcag cctctacatt cacctttgat gattttgcca tgcactgggt ccgtcaagct 120ccagggaagg gtctggagtg ggtctctctt attactgggg atggtggtag cacatactat 180gcagaccctg tgaagggccg attcaccatc tccagagaca acagcaaaaa ctccctgtat 240ctgcaaatga acagtctgag aactgaggac accgccttgt attactgtgt aaaagattgg 300tggatagcag ctcgtccgga ctactactac tacggtatgg acgtctgggg ccaagggacc 360acggtcaccg tctcctca 378521126PRTArtificial SequenceSynthetic 521Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Thr Phe Thr Phe Asp Asp Phe 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Leu Ile Thr Gly Asp Gly Gly Ser Thr Tyr Tyr Ala Asp Pro Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Ser Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95Val Lys Asp Trp Trp Ile Ala Ala Arg Pro Asp Tyr Tyr Tyr Tyr Gly 100 105 110Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 12552224DNAArtificial SequenceSynthetic 522acattcacct ttgatgattt tgcc 245238PRTArtificial SequenceSynthetic 523Thr Phe Thr Phe Asp Asp Phe Ala1 552424DNAArtificial SequenceSynthetic 524attactgggg atggtggtag caca 245258PRTArtificial SequenceSynthetic 525Ile Thr Gly Asp Gly Gly Ser Thr1 552657DNAArtificial SequenceSynthetic 526gtaaaagatt ggtggatagc agctcgtccg gactactact actacggtat ggacgtc 5752719PRTArtificial SequenceSynthetic 527Val Lys Asp Trp Trp Ile Ala Ala Arg Pro Asp Tyr Tyr Tyr Tyr Gly1 5 10 15Met Asp Val528378DNAArtificial SequenceSynthetic 528caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cctcggagac cctgtccatc 60acctgcactg tctctggtgg ctccttcagt agtcacttct ggacctggat ccggcagccc 120ccaggaaagg gactggaatg gattggatat ctccattata gtgggggcac cagctacaac 180ccctccctca agagtcgagt catcatatca gtggacacgt ccaagaacca gttctccctg 240aaactgaact ctgtgaccgc tgcggacacg gccgtgtatt actgtgcgag agctagatcg 300gggattactt ttgggggact tatcgtccct ggttcttttg atatctgggg ccaagggaca 360atggtcaccg tctcttca 378529126PRTArtificial SequenceSynthetic 529Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Ile Thr Cys Thr Val Ser Gly Gly Ser Phe Ser Ser His 20 25 30Phe Trp Thr Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Tyr Leu His Tyr Ser Gly Gly Thr Ser Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Ile Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys Leu Asn Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Ala Arg Ser Gly Ile Thr Phe Gly Gly Leu Ile Val Pro Gly Ser 100 105 110Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 12553024DNAArtificial SequenceSynthetic 530ggtggctcct tcagtagtca cttc 245318PRTArtificial SequenceSynthetic 531Gly Gly Ser Phe Ser Ser His Phe1 553221DNAArtificial SequenceSynthetic 532ctccattata gtgggggcac c 215337PRTArtificial SequenceSynthetic 533Leu His Tyr Ser Gly Gly Thr1 553460DNAArtificial SequenceSynthetic 534gcgagagcta gatcggggat tacttttggg ggacttatcg tccctggttc ttttgatatc 6053520PRTArtificial SequenceSynthetic 535Ala Arg Ala Arg Ser Gly Ile Thr Phe Gly Gly Leu Ile Val Pro Gly1 5 10 15Ser Phe Asp Ile 20536324DNAArtificial SequenceSynthetic 536gaaatagttt tgacacagag tcccggcaca ctgtcactct ctcccgggga aagagccacc 60ttgtcatgta gagcaagtca gtcagtctct agctcttatc tcgcctggta ccagcagaag 120ccgggacagg cccctagact gctgatctac ggggcaagtt ccagggccac cggaatcccc 180gaccggttca gtggaagcgg aagcggaacc gattttactt tgacgatttc tagactggag 240ccagaggatt tcgccgttta ctattgtcaa cagtacggaa gcagcccgtg gacgtttggc 300cagggcacga aggtagaaat caag 324537108PRTArtificial SequenceSynthetic 537Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro 85 90 95Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 10553821DNAArtificial SequenceSynthetic 538cagtcagtct ctagctctta t 215397PRTArtificial SequenceSynthetic 539Gln Ser Val Ser Ser Ser Tyr1 55409DNAArtificial SequenceSynthetic 540ggggcaagt 95413PRTArtificial SequenceSynthetic 541Gly Ala Ser154227DNAArtificial SequenceSynthetic 542caacagtacg gaagcagccc gtggacg 275439PRTArtificial SequenceSynthetic 543Gln Gln Tyr Gly Ser Ser Pro Trp Thr1 5544378DNAArtificial SequenceSynthetic 544caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60atttgttctg tctctggtgg ctccttcagt agtcacttct ggagttggat ccggcagccc 120ccagggaagg gactggagtg gattgggtat gtcctttaca gtgggggcac caattacaac 180ccctccctca agagtcgagt caccatatca gtagacacgt ccaagaatca gttcttcctg 240aaactgagct ctgtgaccgc tgcggacacg gccgattatt actgtgcgag agctatatcg 300gggattacgt ttgggggaat tatcgtccct ggttcttttg atatctgggg ccaagggaca 360atggtcaccg tctcttca 378545126PRTArtificial SequenceSynthetic 545Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Ile Cys Ser Val Ser Gly Gly Ser Phe Ser Ser His 20 25 30Phe Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Tyr Val Leu Tyr Ser Gly Gly Thr Asn Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Phe Leu65 70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Asp Tyr Tyr Cys Ala 85 90 95Arg Ala Ile Ser Gly Ile Thr Phe Gly Gly Ile Ile Val Pro Gly Ser 100 105 110Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 12554624DNAArtificial SequenceSynthetic 546ggtggctcct tcagtagtca cttc 245478PRTArtificial SequenceSynthetic 547Gly Gly Ser Phe Ser Ser His Phe1 554821DNAArtificial SequenceSynthetic 548gtcctttaca gtgggggcac c 215497PRTArtificial SequenceSynthetic 549Val Leu Tyr Ser Gly Gly Thr1 555060DNAArtificial SequenceSynthetic 550gcgagagcta tatcggggat tacgtttggg ggaattatcg tccctggttc ttttgatatc 6055124PRTArtificial SequenceSynthetic 551Ala Arg Ala Ile Ser Gly Ile Thr Phe Gly Gly Ile Ile Val Pro Gly1 5 10 15Ser Phe Asp Ile Ser Pro Gly Lys 20552378DNAArtificial SequenceSynthetic 552caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cctcggagac cctgtccctc 60acctgcactg tctctggtgg ctccttcagt agtcacttct ggagctggat ccggcagccc 120ccagggaagg gactggagtg gattggatat atcttataca ctgggggcac cagcttcaac 180ccctccctca agagtcgagt ctccatgtca gtgggcacgt ccaagaacca gttctccctg 240aaattgagct ctgtgaccgc tgcggacacg gccgtatatt actgtgcgag agctagatcg 300gggataacgt ttacgggtat tatcgtccct ggctcttttg atatctgggg ccaagggaca 360atggtcaccg tctcttca 378553126PRTArtificial SequenceSynthetic 553Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Phe Ser Ser His 20 25 30Phe Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Tyr Ile Leu Tyr Thr Gly Gly Thr Ser Phe Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Ser Met Ser Val Gly Thr Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Ala Arg Ser Gly Ile Thr Phe Thr Gly Ile Ile Val Pro Gly Ser 100 105 110Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 12555424DNAArtificial SequenceSynthetic 554ggtggctcct tcagtagtca cttc 245558PRTArtificial SequenceSynthetic 555Gly Gly Ser Phe Ser Ser His Phe1 555621DNAArtificial SequenceSynthetic 556atcttataca ctgggggcac c 215577PRTArtificial SequenceSynthetic 557Ile Leu Tyr Thr Gly Gly Thr1 555860DNAArtificial SequenceSynthetic 558gcgagagcta gatcggggat aacgtttacg ggtattatcg tccctggctc ttttgatatc 6055920PRTArtificial SequenceSynthetic 559Ala Arg Ala Arg Ser Gly Ile Thr Phe Thr Gly Ile Ile Val Pro Gly1 5 10 15Ser Phe Asp Ile 20560378DNAArtificial SequenceSynthetic 560caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60acttgttctg tctctggtgg ctccttcagt agtcacttct ggagctggat ccggcagccc 120ccagggaagg gactggagtg gattggatat atccattaca gtgggggcac caattacaac 180ccctccctca agagtcgagt caccatatca gtagacacgt ccaagaacca gttctccctt 240aaactgactt ctgtgaccgc tgcggacacg gccgattatt actgtgcgag agctatatcg 300gggattacgt ttgggggaat gatcgtccct ggttcttttg atgtctgggg cgaagggaca 360atggtcaccg tctcttca 378561126PRTArtificial SequenceSynthetic 561Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Ser Val Ser Gly Gly Ser Phe Ser Ser His 20 25 30Phe Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Tyr Ile His Tyr Ser Gly Gly Thr Asn Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys Leu Thr Ser Val Thr Ala Ala Asp Thr Ala Asp Tyr Tyr Cys Ala 85 90 95Arg Ala Ile Ser Gly Ile Thr Phe Gly Gly Met Ile Val Pro Gly Ser 100 105 110Phe Asp Val Trp Gly Glu Gly Thr Met Val Thr Val Ser Ser 115 120 12556224DNAArtificial SequenceSynthetic 562ggtggctcct tcagtagtca cttc 245638PRTArtificial SequenceSynthetic 563Gly Gly Ser Phe Ser Ser His Phe1 556421DNAArtificial SequenceSynthetic 564atccattaca gtgggggcac c 215657PRTArtificial SequenceSynthetic 565Ile His Tyr Ser Gly Gly Thr1 556660DNAArtificial SequenceSynthetic 566gcgagagcta tatcggggat tacgtttggg ggaatgatcg tccctggttc ttttgatgtc 6056720PRTArtificial SequenceSynthetic 567Ala Arg Ala Ile Ser Gly Ile Thr Phe Gly Gly Met Ile Val Pro Gly1 5 10 15Ser Phe Asp Val 20568378DNAArtificial SequenceSynthetic 568caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60acctgcactg tctctggtgg caccttcagt agtcacttct ggagctggat ccggcagccc 120ccaggaaagg gactggagtg gattggatat atcttttaca ctgggggcac caaccacaac 180ccctccctca agagtcgagt caccatatca atagacacgt ccaagaacca gttctccctg 240aaactgacct ctgtgaccgc tgcggacacg gccgtgtatt actgtgcgag agctagatcg 300gggattacgt ttgggggagt tatcgtccct ggttcttttg atatctgggg ccaagggaca 360atggtcaccg tctcttca 378569126PRTArtificial SequenceSynthetic 569Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Thr Phe Ser Ser His 20 25 30Phe Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Tyr Ile Phe Tyr Thr Gly Gly Thr Asn His Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr Ile Ser Ile Asp Thr Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys Leu Thr Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Ala Arg Ser Gly Ile Thr Phe Gly Gly Val Ile Val Pro Gly Ser 100 105 110Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 12557024DNAArtificial SequenceSynthetic 570ggtggcacct tcagtagtca cttc 245718PRTArtificial SequenceSynthetic 571Gly Gly Thr Phe Ser Ser His Phe1 557221DNAArtificial SequenceSynthetic 572atcttttaca ctgggggcac c 215737PRTArtificial SequenceSynthetic 573Ile Phe Tyr Thr Gly Gly Thr1 557460DNAArtificial SequenceSynthetic 574gcgagagcta gatcggggat tacgtttggg ggagttatcg tccctggttc ttttgatatc 6057520PRTArtificial SequenceSynthetic 575Ala Arg Ala Arg Ser Gly Ile Thr Phe Gly Gly Val Ile Val Pro Gly1 5 10 15Ser Phe Asp Ile 20576378DNAArtificial SequenceSynthetic 576caggtgcagc tgcaggagtc gggcccagga ctggtgaaac cttcggagac cctgtccctc 60acctgcactg tctctggtgg ctccttcagc agtcacttct ggaactggat ccggcagtcc 120ccagggaggg gactggaatg gattggatat atctattaca gtgggggcac caactataac 180ccctccttca agagtcgagt caccatgtca gtagacacgt ccaagaacca gttctccctg 240aaactgagct ctgtgaccgc tgcggacacg gccgtgtttt actgtgcgag agctagatcg 300gggataacgt ttgggggagt tctcgtccct ggttcttttg atatttgggg ccaagggaca 360atggtcaccg tctcttca 378577126PRTArtificial SequenceSynthetic 577Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Phe Ser Ser His 20 25 30Phe Trp Asn Trp Ile Arg Gln Ser Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45Gly Tyr Ile Tyr Tyr Ser Gly Gly Thr Asn Tyr Asn Pro Ser Phe Lys 50 55 60Ser Arg Val Thr Met Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Phe Tyr Cys Ala 85 90 95Arg Ala Arg Ser Gly Ile Thr Phe Gly Gly Val Leu Val Pro Gly Ser 100 105 110Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 12557824DNAArtificial SequenceSynthetic 578ggtggctcct tcagcagtca cttc 245798PRTArtificial SequenceSynthetic 579Gly Gly Ser Phe Ser Ser His Phe1 558021DNAArtificial SequenceSynthetic 580atctattaca gtgggggcac c 215817PRTArtificial SequenceSynthetic 581Ile Tyr Tyr Ser Gly Gly Thr1 558260DNAArtificial SequenceSynthetic 582gcgagagcta gatcggggat aacgtttggg ggagttctcg tccctggttc ttttgatatt 6058320PRTArtificial SequenceSynthetic 583Ala Arg Ala Arg Ser Gly Ile Thr Phe Gly Gly Val Leu Val Pro Gly1 5 10 15Ser

Phe Asp Ile 20584378DNAArtificial SequenceSynthetic 584caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60acctgcactg tctctggtgg ctccttcagt agtcacttct ggagctggat ccggcagccc 120ccaggaaagg gactggagtg gattgggtat atctattaca gtgggggcac ccactacaac 180ccctccctcg agagtcgagt caccatatca gtagacacgt ccaagaacca gttctccctg 240aaactgaact ctgtgaccgc tgcggacacg gccgtttatt actgtgcgag agctagatcg 300gggattactt ttgggggact tatcgtccct ggttcttttg atatctgggg ccaagggaca 360atggtcaccg tctcttca 378585130PRTArtificial SequenceSynthetic 585Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Phe Ser Ser His 20 25 30Phe Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Tyr Ile Tyr Tyr Ser Gly Gly Thr His Tyr Asn Pro Ser Leu Glu 50 55 60Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys Leu Asn Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Ala Arg Ser Gly Ile Thr Phe Gly Gly Leu Ile Val Pro Gly Ser 100 105 110Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Ser Pro 115 120 125Gly Lys 13058624DNAArtificial SequenceSynthetic 586ggtggctcct tcagtagtca cttc 245878PRTArtificial SequenceSynthetic 587Gly Gly Ser Phe Ser Ser His Phe1 558821DNAArtificial SequenceSynthetic 588atctattaca gtgggggcac c 215897PRTArtificial SequenceSynthetic 589Ile Tyr Tyr Ser Gly Gly Thr1 559060DNAArtificial SequenceSynthetic 590gcgagagcta gatcggggat tacttttggg ggacttatcg tccctggttc ttttgatatc 6059120PRTArtificial SequenceSynthetic 591Ala Arg Ala Arg Ser Gly Ile Thr Phe Gly Gly Leu Ile Val Pro Gly1 5 10 15Ser Phe Asp Ile 20592375DNAArtificial SequenceSynthetic 592cgggtgcaac tggtgcagtc tgggtctgag gtgaagaagc ctggggcctc agtgaaggtc 60tcctgcaggg cttctggata catcttcacc agttatgata tcaattgggt gcgacaggcc 120actggacaag ggcttgagtg gatgggatgg atgaacccta ataatggtaa cacagcctat 180acacagaagt tccagggcag agtcaccatg accaggaaca cctccataag cacagcctac 240atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagaaaggga 300ttactatggt tcgggaagtt attagggtac ggtatggacg tctggggcca agggaccacg 360gtcaccgtct cctca 375593125PRTArtificial SequenceSynthetic 593Arg Val Gln Leu Val Gln Ser Gly Ser Glu Val Lys Lys Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Arg Ala Ser Gly Tyr Ile Phe Thr Ser Tyr 20 25 30Asp Ile Asn Trp Val Arg Gln Ala Thr Gly Gln Gly Leu Glu Trp Met 35 40 45Gly Trp Met Asn Pro Asn Asn Gly Asn Thr Ala Tyr Thr Gln Lys Phe 50 55 60Gln Gly Arg Val Thr Met Thr Arg Asn Thr Ser Ile Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Lys Gly Leu Leu Trp Phe Gly Lys Leu Leu Gly Tyr Gly Met 100 105 110Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 12559424DNAArtificial SequenceSynthetic 594ggatacatct tcaccagtta tgat 245958PRTArtificial SequenceSynthetic 595Gly Tyr Ile Phe Thr Ser Tyr Asp1 559624DNAArtificial SequenceSynthetic 596atgaacccta ataatggtaa caca 245978PRTArtificial SequenceSynthetic 597Met Asn Pro Asn Asn Gly Asn Thr1 559854DNAArtificial SequenceSynthetic 598gcgagaaagg gattactatg gttcgggaag ttattagggt acggtatgga cgtc 5459918PRTArtificial SequenceSynthetic 599Ala Arg Lys Gly Leu Leu Trp Phe Gly Lys Leu Leu Gly Tyr Gly Met1 5 10 15Asp Val600324DNAArtificial SequenceSynthetic 600gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 60ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 120cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 180gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 240cctgaagatt ttgcagtgta ttactgtcag cagtatggta gctcaccttg gacgttcggc 300caagggacca aggtggaaat caaa 324601108PRTArtificial SequenceSynthetic 601Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro 85 90 95Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 10560221DNAArtificial SequenceSynthetic 602cagagtgtta gcagcagcta c 216037PRTArtificial SequenceSynthetic 603Gln Ser Val Ser Ser Ser Tyr1 56049DNAArtificial SequenceSynthetic 604ggtgcatcc 96053PRTArtificial SequenceSynthetic 605Gly Ala Ser160627DNAArtificial SequenceSynthetic 606cagcagtatg gtagctcacc ttggacg 276079PRTArtificial SequenceSynthetic 607Gln Gln Tyr Gly Ser Ser Pro Trp Thr1 5608120PRTArtificial SequenceSynthetic 608Glu Val Gln Val Leu Glu Ser Gly Gly Asp Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ala Tyr 20 25 30Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Ala Ile Ser Gly Ser Gly Gly Ser Ala Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Lys Asp Gly Ala Trp Lys Met Ser Gly Leu Asp Val Trp Gly Gln 100 105 110Gly Thr Thr Val Ile Val Ser Ser 115 1206098PRTArtificial SequenceSynthetic 609Gly Phe Thr Phe Ser Ala Tyr Ala1 56108PRTArtificial SequenceSynthetic 610Ile Ser Gly Ser Gly Gly Ser Ala1 561113PRTArtificial SequenceSynthetic 611Ala Lys Asp Gly Ala Trp Lys Met Ser Gly Leu Asp Val1 5 10612107PRTArtificial SequenceSynthetic 612Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Asp Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ile Pro Arg Leu Leu Ile 35 40 45Tyr Thr Thr Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Arg Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Val Ala Thr Tyr Tyr Cys Gln Lys Tyr Asp Ser Ala Pro Leu 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 1056136PRTArtificial SequenceSynthetic 613Gln Asp Ile Ser Asp Tyr1 56143PRTArtificial SequenceSynthetic 614Thr Thr Ser16159PRTArtificial SequenceSynthetic 615Gln Lys Tyr Asp Ser Ala Pro Leu Thr1 5616109PRTHomo sapiens 616Asp Phe Gly Leu Asp Cys Asp Glu His Ser Thr Glu Ser Arg Cys Cys1 5 10 15Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile 20 25 30Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu 35 40 45Phe Val Phe Leu Gln Lys Tyr Pro His Thr His Leu Val His Gln Ala 50 55 60Asn Pro Arg Gly Ser Ala Gly Pro Cys Cys Thr Pro Thr Lys Met Ser65 70 75 80Pro Ile Asn Met Leu Tyr Phe Asn Gly Lys Glu Gln Ile Ile Tyr Gly 85 90 95Lys Ile Pro Ala Met Val Val Asp Arg Cys Gly Cys Ser 100 105617426PRTHomo sapiens 617Met Pro Leu Leu Trp Leu Arg Gly Phe Leu Leu Ala Ser Cys Trp Ile1 5 10 15Ile Val Arg Ser Ser Pro Thr Pro Gly Ser Glu Gly His Ser Ala Ala 20 25 30Pro Asp Cys Pro Ser Cys Ala Leu Ala Ala Leu Pro Lys Asp Val Pro 35 40 45Asn Ser Gln Pro Glu Met Val Glu Ala Val Lys Lys His Ile Leu Asn 50 55 60Met Leu His Leu Lys Lys Arg Pro Asp Val Thr Gln Pro Val Pro Lys65 70 75 80Ala Ala Leu Leu Asn Ala Ile Arg Lys Leu His Val Gly Lys Val Gly 85 90 95Glu Asn Gly Tyr Val Glu Ile Glu Asp Asp Ile Gly Arg Arg Ala Glu 100 105 110Met Asn Glu Leu Met Glu Gln Thr Ser Glu Ile Ile Thr Phe Ala Glu 115 120 125Ser Gly Thr Ala Arg Lys Thr Leu His Phe Glu Ile Ser Lys Glu Gly 130 135 140Ser Asp Leu Ser Val Val Glu Arg Ala Glu Val Trp Leu Phe Leu Lys145 150 155 160Val Pro Lys Ala Asn Arg Thr Arg Thr Lys Val Thr Ile Arg Leu Phe 165 170 175Gln Gln Gln Lys His Pro Gln Gly Ser Leu Asp Thr Gly Glu Glu Ala 180 185 190Glu Glu Val Gly Leu Lys Gly Glu Arg Ser Glu Leu Leu Leu Ser Glu 195 200 205Lys Val Val Asp Ala Arg Lys Ser Thr Trp His Val Phe Pro Val Ser 210 215 220Ser Ser Ile Gln Arg Leu Leu Asp Gln Gly Lys Ser Ser Leu Asp Val225 230 235 240Arg Ile Ala Cys Glu Gln Cys Gln Glu Ser Gly Ala Ser Leu Val Leu 245 250 255Leu Gly Lys Lys Lys Lys Lys Glu Glu Glu Gly Glu Gly Lys Lys Lys 260 265 270Gly Gly Gly Glu Gly Gly Ala Gly Ala Asp Glu Glu Lys Glu Gln Ser 275 280 285His Arg Pro Phe Leu Met Leu Gln Ala Arg Gln Ser Glu Asp His Pro 290 295 300His Arg Arg Arg Arg Arg Gly Leu Glu Cys Asp Gly Lys Val Asn Ile305 310 315 320Cys Cys Lys Lys Gln Phe Phe Val Ser Phe Lys Asp Ile Gly Trp Asn 325 330 335Asp Trp Ile Ile Ala Pro Ser Gly Tyr His Ala Asn Tyr Cys Glu Gly 340 345 350Glu Cys Pro Ser His Ile Ala Gly Thr Ser Gly Ser Ser Leu Ser Phe 355 360 365His Ser Thr Val Ile Asn His Tyr Arg Met Arg Gly His Ser Pro Phe 370 375 380Ala Asn Leu Lys Ser Cys Cys Val Pro Thr Lys Leu Arg Pro Met Ser385 390 395 400Met Leu Tyr Tyr Asp Asp Gly Gln Asn Ile Ile Lys Lys Asp Ile Gln 405 410 415Asn Met Ile Val Glu Glu Cys Gly Cys Ser 420 425618340PRTArtificial SequenceSynthetic 618Glu Ala Glu Thr Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu1 5 10 15Glu Arg Thr Asn Gln Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp 20 25 30Lys Arg Leu His Cys Tyr Ala Ser Trp Ala Asn Ser Ser Gly Thr Ile 35 40 45Glu Leu Val Lys Lys Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp 50 55 60Arg Gln Glu Cys Val Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys65 70 75 80Cys Cys Glu Gly Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu 85 90 95Ala Gly Gly Pro Glu Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Ser 100 105 110Gly Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 115 120 125Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 130 135 140Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser145 150 155 160His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 165 170 175Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 180 185 190Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 195 200 205Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 210 215 220Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln225 230 235 240Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 245 250 255Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 260 265 270Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 275 280 285Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 290 295 300Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val305 310 315 320Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 325 330 335Ser Pro Gly Lys 340619411PRTHomo sapiens 619Met Asp Gly Leu Pro Gly Arg Ala Leu Gly Ala Ala Cys Leu Leu Leu1 5 10 15Leu Ala Ala Gly Trp Leu Gly Pro Glu Ala Trp Gly Ser Pro Thr Pro 20 25 30Pro Pro Thr Pro Ala Ala Gln Pro Pro Pro Pro Pro Pro Gly Ser Pro 35 40 45Gly Gly Ser Gln Asp Thr Cys Thr Ser Cys Gly Gly Phe Arg Arg Pro 50 55 60Glu Glu Leu Gly Arg Val Asp Gly Asp Phe Leu Glu Ala Val Lys Arg65 70 75 80His Ile Leu Ser Arg Leu Gln Met Arg Gly Arg Pro Asn Ile Thr His 85 90 95Ala Val Pro Lys Ala Ala Met Val Thr Ala Leu Arg Lys Leu His Ala 100 105 110Gly Lys Val Arg Glu Asp Gly Arg Val Glu Ile Pro His Leu Asp Gly 115 120 125His Ala Ser Pro Gly Ala Asp Gly Gln Glu Arg Val Ser Glu Ile Ile 130 135 140Ser Phe Ala Glu Thr Asp Gly Leu Ala Ser Ser Arg Val Arg Leu Tyr145 150 155 160Phe Phe Ile Ser Asn Glu Gly Asn Gln Asn Leu Phe Val Val Gln Ala 165 170 175Ser Leu Trp Leu Tyr Leu Lys Leu Leu Pro Tyr Val Leu Glu Lys Gly 180 185 190Ser Arg Arg Lys Val Arg Val Lys Val Tyr Phe Gln Glu Gln Gly His 195 200 205Gly Asp Arg Trp Asn Met Val Glu Lys Arg Val Asp Leu Lys Arg Ser 210 215 220Gly Trp His Thr Phe Pro Leu Thr Glu Ala Ile Gln Ala Leu Phe Glu225 230 235 240Arg Gly Glu Arg Arg Leu Asn Leu Asp Val Gln Cys Asp Ser Cys Gln 245 250 255Glu Leu Ala Val Val Pro Val Phe Val Asp Pro Gly Glu Glu Ser His 260 265 270Arg Pro Phe Val Val Val Gln Ala Arg Leu Gly Asp Ser Arg His Arg 275 280 285Ile Arg Lys Arg Gly Leu Glu Cys Asp Gly Arg Thr Asn Leu Cys Cys 290 295 300Arg Gln Gln Phe Phe Ile Asp Phe Arg Leu Ile Gly Trp Asn Asp Trp305 310 315 320Ile Ile Ala Pro Thr Ser Tyr Tyr Gly Asn Tyr Cys Glu Gly Ser Cys 325 330 335Pro Ala Tyr Leu Ala Gly Val Pro Gly Ser Ala Ser Ser Phe His Thr 340 345 350Ala Val Val Asn Gln Tyr Arg Met Arg Gly Leu Asn Pro Gly Thr Val 355 360 365Asn Ser Cys Cys Ile Pro Thr Lys Leu Ser Thr Met Ser Met Leu Tyr 370 375 380Phe Asp Asp Glu Tyr Asn Ile Val Lys Arg Asp Val Pro Asn Met Ile385 390 395 400Val Glu Glu Cys

Gly Cys Ala Ser Pro Gly Lys 405 410620122PRTHomo sapiens 620Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe 20 25 30Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Thr Ile Ser Gly Ser Gly Gly Tyr Thr Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Lys Asp Gly Arg Tyr Asn Trp Asn Tyr Gly Ala Phe Asp Ile Trp 100 105 110Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120621107PRTHomo sapiens 621Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Leu 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105622115PRTArtificial SequenceSynthetic 12A5-5HC polypeptideMOD_RES(50)..(50)Any naturally occurring amino acidMOD_RES(50)..(50)Xaa = Any naturally occurring amino acid 622Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr 20 25 30Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Xaa Ile Arg Leu Lys Ser Asp Asn Tyr Ala Thr His Tyr Ala Glu 50 55 60Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser65 70 75 80Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95Tyr Cys Thr Glu Gly Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr 100 105 110Val Ser Ser 115623107PRTArtificial SequenceSynthetic 12A5-5LC polypeptide 623Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile Asn Lys Tyr 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Tyr Thr Ser Trp Leu Gln Pro Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Ile Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Asn Leu Leu Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105624124PRTArtificial SequenceAntibody Fragment 624Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Gly Tyr 20 25 30Phe Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45Gly Asn Ile Tyr Pro Tyr Asn Ser Val Ser Asn Tyr Asn Gln Arg Phe 50 55 60Lys Ala Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75 80Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Arg Tyr Tyr Tyr Gly Thr Gly Pro Ala Asp Trp Tyr Tyr Asp 100 105 110Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120625107PRTArtificial SequenceAntibody Fragment 625Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile Asn Ser Tyr 20 25 30Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Thr Leu Ile 35 40 45Tyr Arg Ala Asn Arg Leu Val Asp Gly Val Pro Ser Lys Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Ser Asp Glu Phe Pro Leu 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105626117PRTHomo sapiens 626Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45Gly Ile Ile Asn Pro Ser Gly Gly Ser Thr Ser Tyr Ala Gln Lys Phe 50 55 60Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Asp Glu Asn Trp Gly Phe Asp Pro Trp Gly Gln Gly Thr Leu 100 105 110Val Thr Val Ser Ser 115627105PRTHomo sapiens 627Ser Tyr Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ser Pro Gly Gln1 5 10 15Thr Ala Thr Ile Thr Cys Ser Gly His Ala Leu Gly Asp Lys Phe Val 20 25 30Ser Trp Tyr Gln Gln Gly Ser Gly Gln Ser Pro Val Leu Val Ile Tyr 35 40 45Asp Asp Thr Gln Arg Pro Ser Gly Ile Pro Gly Arg Phe Ser Gly Ser 50 55 60Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Thr Gln Ala Met65 70 75 80Asp Glu Ala Asp Tyr Phe Cys Gln Ala Trp Asp Ser Ser Phe Val Phe 85 90 95Gly Gly Gly Thr Lys Val Thr Val Leu 100 105628120PRTHomo sapiens 628Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30Trp Met Gln Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45Gly Ala Ile Tyr Pro Gly Asp Gly Asp Thr Arg Tyr Thr Gln Lys Phe 50 55 60Lys Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Met Gly Gly Tyr Asp Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln 100 105 110Gly Thr Leu Val Thr Val Ser Ser 115 120629113PRTHomo sapiens 629Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly1 5 10 15Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30Ala Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45Pro Pro Lys Leu Leu Ile Tyr Phe Ala Ser Thr Arg Glu Ser Gly Val 50 55 60Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr65 70 75 80Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln 85 90 95His Tyr Asn Thr Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 100 105 110Lys630113PRTHomo sapiensMISC_FEATURE(1)..(1)Xaa is an unknown amino acidMISC_FEATURE(113)..(113)Xaa is an unknown amino acid 630Xaa Val Lys Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Phe 20 25 30Tyr Met Tyr Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45Gly Glu Ile Asn Pro Ser Asn Gly Asp Thr Asn Phe Ile Glu Ser Phe 50 55 60Lys Ser Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr65 70 75 80Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95Thr Val Arg Phe Ala Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser 100 105 110Xaa631112PRTArtificial SequenceSynthetic polypeptide 631Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Thr Ile Ser Ser Gly Gly Ser Tyr Thr Ser Tyr Pro Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Lys Gln Asp Tyr Ala Met Asn Tyr Trp Gly Gln Gly Thr Met Val 100 105 110632107PRTArtificial SequenceSynthetic polypeptide 632Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Thr Ala 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Trp 85 90 95Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105633116PRTArtificial SequenceMouse CDRs and human VH & J segment 633Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Thr Ile Ser Ser Gly Gly Ser Tyr Thr Ser Tyr Pro Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Lys Gln Asp Tyr Ala Met Asn Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110Thr Val Ser Ser 115634107PRTArtificial SequenceMouse CDRs and human VH & J segment 634Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Thr Ala 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Trp 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105635124PRTArtificial SequenceSynthetic construct 635Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Arg Lys Val 20 25 30Gly Ser Ser Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 40 45Trp Ile Gly His Ile Tyr Trp Asp Asp Asp Lys Arg Leu Asn Pro Ser 50 55 60Leu Arg Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe65 70 75 80Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 90 95Cys Ala Arg Arg Ala Ile Thr Thr Val Ile Gly Gly Gly Thr Phe Asp 100 105 110Leu Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120636106PRTArtificial SequenceSynthetic construct 636Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Ile Ser Tyr Met 20 25 30His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45Asp Thr Ser Lys Leu Ala Arg Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu65 70 75 80Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Trp Tyr Leu Asn Pro Leu Thr 85 90 95Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105637124PRTArtificial SequenceSynthetic construct 637Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln1 5 10 15Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Arg Lys Val 20 25 30Gly Arg Ser Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45Trp Leu Ala His Ile Tyr Trp Asp Asp Asp Lys Arg Tyr Asn Pro Ser 50 55 60Leu Arg Asn Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val65 70 75 80Val Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr 85 90 95Cys Ala Arg Arg Ala Ile Thr Thr Val Ile Gly Gly Gly Thr Phe Asp 100 105 110Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120638106PRTArtificial SequenceSynthetic construct 638Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Ile Ser Tyr Met 20 25 30His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45Asp Thr Ser Lys Leu Ala Arg Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu65 70 75 80Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Trp Tyr Arg Asn Pro Leu Thr 85 90 95Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105639113PRTArtificial SequenceSynthetic construct 639Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Phe Ser Arg Tyr 20 25 30Pro Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Val Trp Val 35 40 45Ser Ala Ile Thr Ser Ser Gly Gly Ser Thr Tyr Tyr Ser Asp Thr Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Leu Pro Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser 100 105 110Ser640112PRTArtificial SequenceSynthetic construct 640Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Ser Ser 20 25 30Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Ser Thr Ser Asn Leu Val Ala Gly Ile Pro Asp Arg

Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln His His Ser Gly Tyr His 85 90 95Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Ser Pro Gly Lys 100 105 110641439PRTArtificial SequenceFull length landogrozumab heavy chain 641Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Phe Ser Arg Tyr 20 25 30Pro Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Val Trp Val 35 40 45Ser Ala Ile Thr Ser Ser Gly Gly Ser Thr Tyr Tyr Ser Asp Thr Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Leu Pro Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser 100 105 110Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser 115 120 125Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp 130 135 140Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr145 150 155 160Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr 165 170 175Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys 180 185 190Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp 195 200 205Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala 210 215 220Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro225 230 235 240Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 245 250 255Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val 260 265 270Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 275 280 285Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 290 295 300Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly305 310 315 320Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 325 330 335Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr 340 345 350Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 355 360 365Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 370 375 380Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr385 390 395 400Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe 405 410 415Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys 420 425 430Ser Leu Ser Leu Ser Leu Gly 435642215PRTArtificial SequenceFull length landogrozumab light chain 642Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Ser Ser 20 25 30Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Ser Thr Ser Asn Leu Val Ala Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln His His Ser Gly Tyr His 85 90 95Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala 100 105 110Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120 125Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser145 150 155 160Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys 195 200 205Ser Phe Asn Arg Gly Glu Cys 210 2156437PRTArtificial SequenceHCVR1 643Gly Leu Thr Phe Ser Arg Tyr1 56446PRTArtificial SequenceHCVR2 644Thr Ser Ser Gly Gly Ser1 56454PRTArtificial SequenceHCVR3 645Leu Pro Asp Tyr16469PRTArtificial SequenceLCVR1 646Ser Ser Val Ser Ser Ser Tyr Leu His1 56477PRTArtificial SequenceLCVR2 647Ser Thr Ser Asn Leu Val Ala1 56489PRTArtificial SequenceLCVR3 648Gln His His Ser Gly Tyr His Phe Thr1 5

Claims

1. A method for altering the body composition of a subject comprising administering a first composition comprising an effective amount of a GDF8 inhibitor and a second composition comprising an effective amount of an Activin A inhibitor to the subject.

2. The method of claim 1, wherein the altering of the body composition comprises inducing a reduction in fat mass in the subject comprising administering the first composition comprising an effective amount of a GDF8 inhibitor and the second composition comprising an effective amount of an Activin A inhibitor to the subject.

3. The method of claim 1 or claim 2, wherein the altering of the body composition comprises inducing an increase in muscle mass in the subject, the method comprising administering the first composition comprising an effective amount of the GDF8 inhibitor and the second composition comprising an effective amount of the Activin A inhibitor.

4. A method for altering the body composition of a subject comprising administering a composition comprising an effective amount of a GDF8 inhibitor and an effective amount of an Activin A inhibitor to the subject.

5. The method of claim 4, wherein the altering of the body composition comprises inducing a reduction in fat mass in a subject comprising administering the composition comprising an effective amount of the GDF8 inhibitor and an effective amount of the Activin A inhibitor to the subject.

6. The method of claim 3 or claim 4, wherein the altering of the body composition comprises inducing an increase in muscle mass in the subject, the method comprising administering to the subject in need thereof the composition comprising an effective amount of the GDF8 inhibitor and an effective amount of the Activin A inhibitor.

7. The method of any one of claims 1-6, wherein the effective amount of a GDF8 inhibitor comprises a dosing regimen selected from the group consisting of at least 0.01 mg/kg to about 10 gm/kg, 1 mg/kg to about 1 gm/kg, and 10 mg/kg to 100 mg/kg.

8. The method of any one of claims 1-7, wherein the effective amount of an Activin A inhibitor comprises a dosing regimen selected from the group consisting of at least 0.01 mg/kg to about 10 gm/kg, 1 mg/kg to about 1 gm/kg, and 10 mg/kg to 100 mg/kg.

9. The method of any one of claims 1-6, wherein the effective amount of a GDF8 inhibitor comprises a dosing regimen selected from a group consisting of a single dose of about 0.01 to about 20 mg/kg body weight, about 0.1 to about 10 mg/kg body weight, and about 0.1 to about 5 mg/kg body weight.

10. The method of any one claim 1-6 or 9, wherein the effective amount of an Activin A inhibitor comprises a dosing regimen selected from a group consisting of a single dose of about 0.01 to about 20 mg/kg body weight, about 0.1 to about 10 mg/kg body weight, and about 0.1 to about 5 mg/kg body weight.

11. The method of any one of claims 7-10, wherein the effective amount of the GDF8 inhibitor is 6 mg/kg body weight of the subject.

12. The method of any one of claims 7-11, wherein the effective amount of the Activin A inhibitor is 3 mg/kg or 10 mg/kg body weight of the subject.

13. The method of any one of claims 1-3, wherein the first composition is formulated for intravenous, subcutaneous, or oral administration.

14. The method of any one of claims 1-3, wherein the second composition is formulated for intravenous, subcutaneous, or oral administration.

15. The method of any one of claims 1-3, wherein the first and second compositions are combined into a third composition prior to administration.

16. The method of claim 15, wherein the third composition is formulated for intravenous, subcutaneous, or oral administration.

17. The method of any one of claims 3-6, wherein the composition is formulated for intravenous, subcutaneous, or oral administration.

18. The method of any one of claims 1-17, further comprising measuring total fat mass and/or android fat mass in the subject before administration.

19. The method of claim 18, further comprising measuring total fat mass and/or android fat mass in the subject after administration, and administering the composition until the subject has a reduction in total fat mass and/or android fat mass of at least about 2% to 8%, at least about 2.5% to 6%, at least about 3% to 4%, or at least about 3.5%.

20. The method of any one of claims 1-19, wherein the GDF8 inhibitor is an isolated antibody or an antigen-binding fragment thereof that specifically binds to GDF8.

21. The method of claim 20, wherein the antibody or antigen-binding fragment that specifically binds GDF8 comprises the heavy chain complementarity determining regions (HCDRs) of a heavy chain variable region (HCVR) comprising SEQ ID NO:360, and the light chain complementarity determining regions (LCDRs) of a light chain variable region (LCVR) comprising SEQ ID NO:368.

22. The method of claim 20 or 21, wherein the antibody or antigen-binding fragment that specifically binds GDF8 comprises three HCDRs comprising SEQ ID NO:362, SEQ ID NO:364, and SEQ ID NO:366, and three LCDRs comprising SEQ ID NO:370, SEQ ID NO:372, and SEQ ID NO:374.

23. The method of any one of claims 1-22, wherein the Activin A inhibitor is an isolated antibody or antigen-binding fragment thereof that specifically binds Activin A.

24. The method of claim 23, wherein the antibody or antigen-binding fragment that specifically binds Activin A comprises the heavy chain complementarity determining regions (HCDRs) of a heavy chain variable region (HCVR) comprising SEQ ID NO:553, and the light chain complementarity determining regions (LCDRs) of a light chain variable region (LCVR) comprising SEQ ID NO:537.

25. The method of claim 23 or 24, wherein the antibody or antigen-binding fragment that specifically binds Activin A comprises three HCDRs comprising SEQ ID NO:555, SEQ ID NO:557, and SEQ ID NO:559, and three LCDRs comprising SEQ ID NO:539, SEQ ID NO:541, and SEQ ID NO:543.

26. The method of any one of claims 1-25, wherein the Activin A inhibitor is in an amount selected from the group consisting of between 100% to 200% of the amount of the GDF8 inhibitor, between 100% and 250% of the amount of the GDF8 inhibitor, between 100% and 300% of the amount of the GDF8 inhibitor, and between 100% and 400% by weight of the amount of the GDF8 inhibitor.

27. The method of claim 26, wherein the amount of the Activin A inhibitor is about 1.5 to 2.0 times as large by weight as the amount of the GDF8 inhibitor.

28. The method of any one of claims 1-27, wherein a reduction of fat mass in the subject is a reduction in total fat mass as measured by DXA (Dual-energy X-ray absorptiometry).

29. The method of any one of claims 1-28, wherein a reduction of fat mass in the subject is a reduction in android fat mass as measured by DXA (Dual-energy X-ray absorptiometry).

30. The method of any one of claims 1-29, wherein the subject experiences an increase in muscle volume after administration.

31. The method of any one of claims 1-30, wherein the subject does not have a muscle wasting condition or disease.

32. A GDF8 inhibitor for use in a method for treating a disease or disorder characterized by increased fat mass, wherein the method comprises administrating to a subject the GDF8 inhibitor and an Activin A inhibitor.

33. An Activin A inhibitor for use in a method for treating a disease or disorder characterized by increased fat mass, wherein the method comprises administrating to a subject the Activin A inhibitor and a GDF8 inhibitor.

34. A non-therapeutic method for decreasing fat mass in a subject, the method comprising administering to the subject an Activin A inhibitor and a GDF8 inhibitor.