Use of Flecainide as an Anti-Connexin Agent and Method for Potentiating the Effects of a Psychotropic Drug

Abstract

The present invention relates to the use of flecainide as an anti-connexin agent. This anti-connexin agent is advantageously used to potentiate the therapeutic effect of various psychotropic drugs. More specifically, the invention provides a combination product containing flecainide and modafinil.

Description

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation of U.S. patent application Ser. No. 14/907,221, filed Jan. 22, 2016, no U.S. Pat. No. 9,750,734, which claims priority to international application no. PCT/EP2014/065975, filed Jul. 24, 2014, which claims priority to EP application no. 13306074.9, filed Jul. 24, 2013, the entire disclosures of which are hereby incorporated by reference.

SUMMARY OF THE INVENTION

[0002] The present invention relates to the use of flecainide as an anti-connexin agent. This anti-connexin agent is advantageously used to potentiate the therapeutic effect of various psychotropic drugs. More specifically, the invention provides a combination product containing flecainide and modafinil.

BACKGROUND OF THE INVENTION

[0003] Gap junctions are involved in intercellular communication, which is important for maintaining tissue and organ homeostasis. Gap junctions connect the cell cytoplasm, enabling the exchange of ions (Ca.sup.+ and K.sup.+), second messengers (AMPc, GMPc, IP3), several small metabolites (glucose) and ensuring electrical and metabolic coupling between the cells. The gap junctions are junctions with a selective permeability, formed by protein channels contained in the plasma membrane, and formed by connexin hexamers. Connexin hexamers might as well form hemichannel, linked the intracellular space to extracellular one.

[0004] Connexins are integral proteins of the plasma membrane, which are synthesized by practically every cell type, regardless of the position of a multicellular organism in the phylogenesis of the animal world. In vertebrates, occasional cells not producing connexins are adult striated muscle cells, spermatozoids and circulating blood cells. Unlike numerous membrane proteins, connexins have a short half-life (between 3 and 6 hours), are not glycosylated and do not have an enzymatic activity. At present, at least thirteen distinct connexins have been identified in mammals; corresponding, in humans, to 21 isoforms. In practice, various types of connexins can be present in a plurality of tissues, and most of the cells synthesize a plurality of connexins. Before reaching the cell membrane, the connexins assemble in groups of six molecules to form hollow tubular structures called connexons, which join the plasma membrane by means of Golgi vesicles. When cell contact is established, the connexons of a cell align end-to-end with those of the neighboring cell, establishing a continuous hydrophilic channel around 10 nm long. This junctional channel establishes direct contact between the cytoplasms of the two cells in contact, over the intercellular space.

[0005] Connexins are involved in a huge number of physiological processes, and several applications of connexin blocking agents (also called hereafter "connexin blocking agents" or "anti-connexin agents") have been described.

[0006] For example, anti-connexin agents have been proposed for treating and/or preventing the following conditions:

[0007] cancers (WO2006/134494 and WO2006/049157),

[0008] some cardiovascular diseases (WO2006/134494),

[0009] wounds (WO2006/134494 and WO2009/097077),

[0010] pain (WO2009/148613),

[0011] migraines (Durham and Garrett, 2009),

[0012] epilepsy (Juszczak and Swiergiel, 2009),

[0013] neurological conditions (WO2006/134494) and neurodegenerative diseases (Takeuchi et al. 2011),

[0014] ischemia (Davidson et al, 2013),

[0015] drug-induced liver injury (Patel et al, 2012)

[0016] infectious diseases (WO2011/067607),

[0017] cytotoxicity induced by chemotherapeutic agents (Tong X. et al, 2013) and

[0018] inflammatory disorders (WO2006/134494).

[0019] Furthermore, the present inventors previously described that anti-connexin agents are able to potentiate the therapeutic effects of psychotropic drugs (WO 2010/029131). In particular, they described that administration of anti-connexin agents such as meclofenamic acid (MFA) increases the therapeutic effects of various psychotropic molecules, enabling to reduce the active doses and thus the undesirable effects of these psychotropic molecules. These synergistic effects have been observed with a wide range of psychotropic molecules (clozapine, paroxetine, modafinil, diazepam, venlafaxine, escitalopram, bupropion and sertraline).

[0020] Identifying new anti-connexin agents is therefore of primary importance to highlight new therapeutic tools aiming to treat various diseases and disorders, in particular in combination with psychotropic drugs.

[0021] In this context, the inventors have now demonstrated that the well-known antiarrhythmic agent flecainide, has a broad anti-connexin activity. This is a very surprising result, since flecainide had been described so far to interfere with sodium channels, in particular on heart muscle cells, and these channels are not related with brain gap junctions. Moreover, flecainide had been shown not to influence junctional resistance of cardiac myocyte cell pairs (Daleau et al, 1998).

DETAILED DESCRIPTION OF THE INVENTION

[0022] In the context of the invention, "flecainide" designates a compound of formula N-(piperidin-2-ylmethyl)-2,5-bis(2,2,2-trifluoroethoxy) benzamide. As used herein, this term designates any form of this compound, such as a salt thereof. Preferably, said salt is the flecainide acetate. This term may also encompass the flecainide precursors which can be metabolized in the human body and/or its derivatives (for example, chemical derivatives resulting from one or several halogen substitutions and/or from addition of protective groups).

[0023] As disclosed on FIGS. 5A and 5B, flecainide possesses a chiral center implying the existence of an R and S enantiomers (S-(+)-flecainide and R-(-)-flecainide). FIGS. 5A-5B show the formulas of R-flecainide (FIG. 5A, (R)--N-(piperidin-2-ylmethyl)-2,5-bis(2,2,2-trifluoroethoxy)ben- zamide) and S-flecainide (FIG. 5B, (S)--N-(piperidin-2-ylmethyl)-2,5-bis(2,2,2-trifluoroethoxy)benzamide).

[0024] As used herein, the term "flecainide" designates the racemate form of N-(piperidin-2-ylmethyl)-2,5-bis(2,2,2-trifluoroethoxy) benzamide, as well as the R and S enantiomers thereof ((R)--N-(piperidin-2-ylmethyl)-2,5-bis(2,2,2-trifluoroethoxy)benzamide and (S)--N-(piperidin-2-ylmethyl)-2,5-bis(2,2,2-trifluoroethoxy)benzamide- , respectively). In a preferred embodiment of the invention, the R enantiomer of flecainide ((R)--N-(piperidin-2-ylmethyl)-2,5-bis(2,2,2-trifluoroethoxy)benzamide) will be used.

[0025] Flecainide is currently administered as a racemate (Kroemer et al, 1989; Lie et al, 1989). The pharmacokinetic parameters of the two enantiomers of flecainide have been largely described, after administration in human and rodents, as described below:

[0026] In 1989, Kroemer et al. published a study in 13 patients receiving long-term oral flecainide therapy. S-flecainide and R-flecainide plasma levels were determined, and plasma concentrations of R-flecainide were significantly higher than those of the S-flecainide enantiomer (R/S ratio=1.10), suggesting that the flecainide drug undergoes modest enantioselective disposition [Kroemer et al, 1989].

[0027] In 1989, Gross et al. compared the disposition of the two enantiomers in two human populations: extensive (EM) and five poor (PM) metabolizers of sparteine/debrisoquine after administration of 50 mg of racemic flecainide acetate [Gross et al, 1989]. Gross et al. presented data indicating that the half-life of R-flecainide (12.9h) was longer (P<0.03) than that of S-flecainide (9.8h). The renal clearance of the two enantiomers was, however, comparable and similar to that observed in the EM subjects. The urinary recovery of R-flecainide (15.6.+-.3.7 mg) was greater (P<0.03) than that of the S-enantiomer (12.0.+-.3.7 mg). The enantioselective disposition observed in PMs is therefore due to greater impairment in the metabolism of R-flecainide than S-flecainide.

[0028] In 1991, Alessi-Severini et al. summarized key findings on pharmacokinetics and concluded that there was no evidence of enantioselective disposition of flecainide in human [Alessi-Severini et al., 1991], citing three reports on stereoselective therapeutic monitoring, which found R/S ratio ranges of 0.67-1.39 (mean 1.03.+-.0.16), 0.75-1.44 (mean 1.04), and 0.89-1.32 (mean 1.10.+-.0.13), and that Gross et al. 1989 study was not relevant on the total population.

[0029] In 1998, Hanada et al. demonstrated an absence of enantioselective distribution of the two enantiomers of flecainide in several tissue, after intravenous administration of flecainide racemate in rats [Hanada et al, 1998].

[0030] As reviewed in [Mehvar et al, 2002], it appears that the renal clearances of the enantiomers of flecainide are not stereoselective in both healthy volunteers and patients.

[0031] Literature is thus globally coherent on the absence of stereoselective effects of flecainide on pharmacokinetics and metabolism.

[0032] The physicochemical properties of the two enantiomers of flecainide have been also described. In particular, Turgeon et al. described a stereoselective analytical method for the determination of the antiarrhythmic agent flecainide in human plasma. The resolution of the enantiomers is achieved by high-performance liquid chromatography (HPLC) on a normal phase silica column following derivatization with the optically active reagent (-)-methyl chloroformate [Turgeon et al., 1990].

[0033] Moreover, Alessi-Severini et al. described a stereospecific high-performance liquid chromatographic method for the determination of (R,S)-flecainide acetate in human plasma and urine. Flecainide diastereomers were separated after i) a single-step extraction of alkalinized samples performed with distilled diethyl ether, ii) the organic layer was evaporated and the drug was derivatized with 1-[(4-nitrophenyl)sulfonyl]-L-propyl chloride at 80 degrees C. for 2 h and iii) by high-performance liquid chromatography (HPLC) on a C18 reversed-phase column with a mobile phase consisting of acetonitrile:water:triethylamine (45:55:0.2) at a flow rate of 1 mL/min [Alessi-Severini et al., 1990].

[0034] Racemic flecainide acetate is a widely used class 1c antiarrhythmic agent, which is indicated for treating various types of arrhythmias. More specifically, it is used to regulate the rate and rhythm of the heart. The heart's pumping action is controlled by electrical signals that pass through the heart muscle. These electrical signals cause the two pairs of heart chambers (left and right arteria and ventricles) to contract in a regular manner to produce regular heartbeats. If the electrical activity in the heart is disturbed for any reason, irregular heartbeats (arrhythmias) of various types can result. Flecainide helps to treat arrhythmias by decreasing the sensitivity of the heart muscle cells to electrical impulses. This regulates the electrical conduction in the heart muscle and reduces disturbances in the heart rhythm. As a class I antiarrhythmic agent, flecainide interferes with the sodium channel.

[0035] Importantly, several studies have demonstrated that these cardiovascular effects are not mediated by a single enantiomer, both of them contributing to cardiovascular functions:

[0036] Antiarrhythmic effects of flecainide and its enantiomers were assessed in two different animal models, chloroform-induced ventricular fibrillation in mice and ouabain-induced ventricular tachycardia in dogs. The two enantiomers were highly effective in suppressing both of these experimental arrhythmias and appeared to be essentially equipotent. No significant differences were found either between the two enantiomers or between the enantiomers and racemic flecainide [Banitt et al, 1986].

[0037] The effects of the enantiomers on action potential characteristics in canine cardiac Purkinje fibers were assessed, and they were shown to exert similar electrophysiological effects [Kroemer et al, 1989].

[0038] The effects of flecainide acetate racemate and its two enantiomers on voltage-operated sodium and potassium channels and on the sodium pump activity of non-myelinated fibers of the guinea-pig vagus nerve were studied with the sucrose-gap method. There was no significant difference in the effect caused by the enantiomers separately [Lie et al, 1989].

[0039] The effects of the enantiomers were evaluated in isolated canine Purkinje fibers using standard microelectrode techniques. The results suggest there is no significant difference between the effects of flecainide enantiomers on basic electro-physiological parameters of canine Purkinje fibers [Smallwood et al, 1989].

[0040] To conclude, all those studies have provided no evidence to indicate that administration of a single enantiomer, rather than the racemic drug, would offer any advantage.

[0041] According to a first aspect, the present invention therefore pertains to the use of flecainide, in vitro and in vivo, as an anti-connexin agent. In particular, the present invention relates to flecainide for use as an anti-connexin agent, or, in other words, for blocking gap junctions.

[0042] There are 21 genes coding for different connexin isoforms in humans, and different combinations of connexin monomers involved in the composition of the gap junctions are described. In particular, the connexins 26 (Cx 26), 30 (Cx 30), 30.2 (Cx30.2), 32 (Cx 32), 36 (Cx 36), 37 (Cx 37), 40 (Cx 40), 43 (Cx 43), 45 (Cx 45), 46 (Cx 46), and 47 (Cx 47) are expressed in human on cells of the Central or Peripheral Nervous System (Nakase & Naus, 2004).

[0043] The present inventors observed that flecainide is effective for inhibiting gap junctions made of all connexin they tested. In particular, and as disclosed in the experimental part below, flecainide is effective for inhibiting gap junctions made of connexin Cx40, Cx26, Cx30, Cx32, and/or Cx43. Importantly, this anti-connexin effect is similar to the one observed for well-known anti-connexin agents such as mefloquine and meclofenamic acid (MFA) (Juszczak & Swiergiel, 2009; Cruikshank et al, 2004; Harks et al, 2001). Higher inhibition levels were even reached for glial connexins Cx26, Cx30 and Cx43 (see FIGS. 1A-B).

[0044] The present invention therefore relates to the in vitro use of flecainide as an anti-connexin agent. Preferably, this agent can be used to inhibit gap junctions made of the connexins selected in the group consisting of: Cx23 (SEQ ID NO:1), Cx25 (SEQ ID NO:2), Cx26 (SEQ ID NO:3), Cx 30 (SEQ ID NO:4), Cx30.2 (SEQ ID NO:5), Cx30.3 (SEQ ID NO:6), Cx31 (SEQ ID NO:7), Cx31.1 (SEQ ID NO:8), Cx31.9 (SEQ ID NO:9), Cx32 (SEQ ID NO:10), Cx36 (SEQ ID NO:11), Cx37 (SEQ ID NO:12), Cx40 (SEQ ID NO:13), Cx40.1 (SEQ ID NO:14), Cx43 (SEQ ID NO:15), Cx45 (SEQ ID NO:16), Cx46 (SEQ ID NO:17), Cx47 (SEQ ID NO:18), Cx50 (SEQ ID NO:19), Cx59 (SEQ ID NO:20), and Cx62 (SEQ ID NO:21).

[0045] In a preferred embodiment of the invention, flecainide is used for blocking one or more of the connexins expressed in human cells of the Central or Peripheral Nervous System, that are selected in the group consisting of: Cx 26 (SEQ ID NO:3), Cx 30 (SEQ ID NO:4), Cx 30.2 (SEQ ID NO:5), Cx 32 (SEQ ID NO:10), Cx 36 (SEQ ID NO:11), Cx 37 (SEQ ID NO:12), Cx 40 (SEQ ID NO:13), Cx 43 (SEQ ID NO:15), Cx 45 (SEQ ID NO:16), Cx 46 (SEQ ID NO:17) and Cx 47 (SEQ ID NO:18).

[0046] In a more preferred embodiment, flecainide is used for blocking one or more of the connexins selected in the group consisting of: Cx40 (SEQ ID NO:13), Cx26 (SEQ ID NO:3), Cx30 (SEQ ID NO:4), Cx32 (SEQ ID NO:10), and Cx43 (SEQ ID NO:15).

[0047] In an even more preferred embodiment, flecainide is used for blocking one or more of the connexins selected in the group consisting of: Cx26 (SEQ ID NO:3), Cx30 (SEQ ID NO:4) and Cx43 (SEQ ID NO:15).

[0048] Due to its anti-connexin activity, flecainide can be used for the treatment of a number of disorders and conditions known to benefit from treatment by anti-connexin molecules.

[0049] These disorders and conditions include, but are not limited to: cancers, cardiovascular diseases, wounds, pain, migraines, epilepsy, neurological conditions and neurodegenerative diseases, infectious diseases, drug-induced liver injury, cytotoxicity induced by chemotherapeutic agents, ischemia and inflammatory disorders.

[0050] More preferably, flecainide can be used for the prevention and/or the treatment of cancers, wounds, migraines, epilepsy, infectious diseases, drug-induced liver injury, cytotoxicity induced by chemotherapeutic agents, ischemia and inflammatory disorders.

[0051] Even more preferably, flecainide can be used for the prevention and/or the treatment of wounds, migraines, infectious diseases, drug-induced liver injury, cytotoxicity induced by chemotherapeutic agents, and ischemia.

[0052] Even more preferably, flecainide can be used for the prevention and/or the treatment of drug-induced liver injury, cytotoxicity induced by chemotherapeutic agents, and ischemia.

[0053] According to a particular aspect of the present invention, flecainide is used as an agent for potentiating the effects of a psychotropic drug. These potentiating effects are illustrated below by experiments performed with modafinil (see FIGS. 2A-B to 4). As an anti-connexin agent, flecainide can potentiate the effects of any psychotropic drug (as shown in WO 2010/029131 and US 2011/172188, incorporated by reference).

[0054] The term "potentiate" in this case means that flecainide significantly increases the therapeutic effects of the psychotropic drug administered to the same patient. Thus, the combination of the psychotropic drug with flecainide makes it possible to reduce the doses of said psychotropic drug and therefore to limit the adverse effects of said psychotropic drug, and/or to obtain a stronger therapeutic effect without increasing the dose of said psychotropic drug.

[0055] In the present text, a "psychotropic drug" or "psychotropic agent" refers to any substance that acts primarily on the state of the central nervous system by modifying certain cerebral biochemical and physiological processes. Examples of psychotropic drugs which can be used in the context of the present invention include anesthetics, analgesics such as opioids, antipyretics and antimigraine preparations, anti-epileptics, anti-Parkinson drugs such as anti-cholinergic and dopaminergic anti-Parkinson agents, psycholeptics such as antipsychotics, anxiolytics, hypnotics and sedatives, psychoanaleptics such as antidepressants, psychostimulants and anti-dementia drugs, as well as parasymptomimetics, anti-addiction drugs, antivertigo preparations etc. Non-limitative examples of specific molecules which can be advantageously used as psychotropic drugs according to the invention are listed in Table 1 below.

TABLE-US-00001 TABLE 1 Psychotropic molecules Therapeutic Pharmacological Chemical category sub-class sub-class Active agent Anesthetics 1. General 2. Ethers 3. diethyl ether; vinyl ether anesthetics 4. Halogenated 5. halothane; chloroform; hydrocarbons enflurane; trichloroethylene; isoflurane; desflurane; sevoflurane 6. Barbiturates, 7. methohexital; hexobarbital; plain 8. Barbiturates 9. narcobarbital in combination with other drugs 10. Opioid 11. fentanyl; alfentanil; anesthetics sufentanil; phenoperidine; anileridine; remifentanil; 12. Other 13. droperidol; ketamine; general propanidid; alfaxalone; etomidate; anesthetics propofol; sodium oxybate; nitrous oxide; esketamine; xenon; 14. Local 15. Esters of 16. metabutethamine; procaine; anesthetics aminobenzoic tetracaine; chloroprocaine; acid benzocaine; 17. Amides 18. bupivacaine; lidocaine; mepivacaine; prilocaine; butanilicaine; cinchocaine; etidocaine; articaine; ropivacaine; levobupivacaine; bupivacaine; 19. Esters of 20. cocaine benzoic acid 21. Other local 22. ethyl chloride; dyclonine; anesthetics phenol; capsaicin Analgesics 23. Opioids 24. Natural 25. opium; hydromorphone; opium alkaloids nicomorphine; oxycodone; dihydrocodeine; diamorphine; papaveretum; morphine; codeine, 26. Phenylpiperidine 27. ketobemidone; pethidine; derivatives 28. Diphenylpropylamine 29. dextromoramide; piritramide; derivatives dextropropoxyphene; bezitramide; methadone, 30. Benzomorphan 31. pentazocine; phenazocine derivatives 32. Morphinan 33. butorphanol; nalbuphine derivatives 34. Other 35. tilidine; tramadol; dezocine; opioids meptazinol; tapentadol; 36. Other 37. Salicylic 38. acetylsalicylic acid; aloxiprin; analgesics and acid and choline salicylate; sodium antipyretics derivatives salicylate; salicylamide; salsalate; ethenzamide; morpholine salicylate; dipyrocetyl; benorilate; diflunisal; potassium salicylate; guacetisal; carbasalate calcium; imidazole salicylate 39. Pyrazolones 40. phenazone; metamizole sodium; aminophenazone; propyphenazone; nifenazone; 41. Anilides 42. paracetamol; phenacetin; bucetin; propacetamol; 43. Other 44. rimazolium; glafenine; analgesics and floctafenine; viminol; nefopam; antipyretics ziconotide; methoxyflurane; nabiximols 45. Antimigraine 46. Ergot 47. Dihydroergotamine; Preparations alkaloids ergotamine; methysergide; lisuride; 48. Corticosteroid 49. flumedroxone derivatives 50. Selective 51. sumatriptan; naratriptan; serotonin (5HT1) zolmitriptan; rizatriptan; agonists almotriptan; eletriptan; frovatriptan 52. Other 53. pizotifen; clonidine; antimigraine iprazochrome; dimetotiazine; preparations oxetorone Anti- 54. Anti- 55. Barbiturates 56. methylphenobarbital; epileptics epileptics and derivatives Phenobarbital; primidone; barbexaclone; metharbital 57. Hydantoin 58. ethotoin; phenytoin; derivatives amino(diphenylhydantoin) valeric acid; mephenytoin; fosphenytoin; 59. Oxazolidine 60. paramethadione; derivatives trimethadione; ethadione 61. Succinimide 62. Ethosuximide; phensuximide; derivatives mesuximide; 63. Benzodiazepine 64. clonazepam derivatives 65. Carboxamide 66. carbamazepine; derivatives oxcarbazepine; rufinamide; eslicarbazepine 67. Fatty acid 68. valproic acid; valpromide; derivatives aminobutyric acid; vigabatrin; progabide; tiagabine 69. Other 70. sultiame; phenacemide; antiepileptics lamotrigine; felbamate; topiramate; gabapentin; pheneturide; levetiracetam; zonisamide; pregabalin; stiripentol; lacosamide; carisbamate; retigabine; beclamide Anti- 71. Anticholinergic 72. Tertiary 73. Trihexyphenidyl; biperiden; Parkinson agents amines metixene; procyclidine; drugs profenamine; dexetimide; phenglutarimide; mazaticol; bornaprine; tropatepine 74. Ethers 75. etanautine; orphenadrine chemically close to antihistamines 76. Ethers of 77. benzatropine; etybenzatropine tropine or tropine derivatives 78. Dopaminergic 79. Dopa and dopa 80. levodopa; decarboxylase agents derivatives inhibitor; COMT inhibitor; melevodopa; etilevodopa 81. Adamantane 82. amantadine derivatives 83. Dopamine 84. bromocriptine; pergolide; agonists dihydroergocryptine; esylate; ropinirole; pramipexole; cabergoline; apomorphine; piribedil; rotigotine 85. Monoamine 86. selegiline; rasagiline oxidase B inhibitors 87. Other 88. olcapone; entacapone; dopaminergic budipine agents Psycho- 89. Antipschotics 90. Phenothiazines 91. chlorpromazine; leptics with aliphatic side- levomepromazine; promazine; chain acepromazine; triflupromazine; cyamemazine; chlorproethazine 92. Phenothiazines 93. dixyrazine; fluphenazine; with perphenazine; prochlorperazine; piperazine thiopropazate; trifluoperazine; structure acetophenazine; thioproperazine; butaperazine; perazine 94. Phenothiazines 95. periciazine; thioridazine; with mesoridazine; pipotiazine piperidine structure 96. Butyrophenone 97. Haloperidol; trifluperidol; derivatives melperone; moperone; pipamperone; bromperidol; benperidol; droperidol; fluanisone 98. Indole 99. oxypertine; molindone; derivatives sertindole; ziprasidone 100. Thioxanthene 101. flupentixol; clopenthixol; derivatives chlorprothixene; tiotixene; zuclopenthixol 102. Diphenylbutyl- 103. fluspirilene; pimozide; piperidine derivatives penfluridol 104. Diazepines, 105. loxapine; clozapine; oxazepines, olanzapine; quetiapine; asenapine; thiazepines and clotiapine oxepines 106. Benzamides 107. sulpiride; sultopride; tiapride; remoxipride; amisulpride; veralipride; levosulpiride 108. Lithium 109. lithium 110. Other 111. prothipendyl; risperidone; antipsychotics mosapramine; zotepine; aripiprazole; paliperidone 112. Anxiolytics 113. Benzodiazepine 114. chlordiazepoxide; derivatives medazepam; oxazepam; potassium clorazepate; lorazepam; adinazolam; bromazepam; clobazam; ketazolam; prazepam; alprazolam; halazepam; pinazepam camazepam; nordazepam; fludiazepam; ethyl loflazepate; etizolam; clotiazepam; cloxazolam; tofisopam; 115. Diphenylmethane 116. hydroxyzine; captodiame; derivatives 117. Carbamates 118. meprobamate; emylcamate; mebutamate; 119. Dibenzo- 120. benzoctamine bicyclo-octadiene derivatives 121. Azaspirodecane- 122. buspirone dione derivatives 123. Other 124. Mephenoxalone; gedocarnil; anxiolytics etifoxine 125. Hypnotics 126. Barbiturates, 127. Pentobarbital; amobarbital; and plain butobarbital; barbital; aprobarbital; sedatives secobarbital; talbutal; vinylbital; vinbarbital; cyclobarbital; heptabarbital; reposal; methohexital; thiopental; etallobarbital; allobarbital; proxibarbal 128. Aldehydes 129. chloral hydrate; chloralodol; and derivatives acetylglycinamide; dichloralphenazone; paraldehyde 130. Benzodiazepine 131. flurazepam; nitrazepam; derivatives flunitrazepam; estazolam; triazolam; lormetazepam; temazepam; midazolam; brotizolam; quazepam; loprazolam; doxefazepam; cinolazepam 132. Piperidinedione 133. glutethimide; methyprylon; derivatives pyrithyldione 134. Benzodiazepine 135. zopiclone; zolpidem; related drugs zaleplon; eszopiclone 136. Melatonin 137. melatonin; ramelteon receptor agonists 138. Other 139. methaqualone; clomethiazole; hypnotics and bromisoval; carbromal; scopolamine; sedatives propiomazine; triclofos ethchlorvynol; valerian; hexapropymate; bromides; apronal; valnoctamide; methylpentynol; niaprazine; dexmedetomidine 140. Hypnotics 141. emepronium; and sedatives in dipiperonylaminoethanol combination, excl. barbiturates Psycho- 142. Antidepressants 143. Non- 144. desipramine; imipramine; analeptics selective imipramine oxide; clomipramine; monoamine opipramol; trimipramine; reuptake lofepramine; dibenzepin; inhibitors amitriptyline; nortriptyline; protriptyline; doxepin; iprindole; melitracen; butriptyline; dosulepin; amoxapine; dimetacrine; amineptine; maprotiline; quinupramine 145. Selective 146. zimeldine; fluoxetine; serotonin reuptake citalopram; paroxetine; sertraline; inhibitors alaproclate; fluvoxamine; etoperidone; escitalopram 147. Monoamine 148. isocarboxazid; nialamide; oxidase inhibitors, phenelzine; tranylcypromine; non-selective iproniazide; iproclozide 149. Monoamine 150. moclobemide; toloxatone oxidase A inhibitors 151. Other 152. oxitriptan; tryptophan; antidepressants mianserin; nomifensine; trazodone; nefazodone; minaprine; bifemelane; viloxazine; oxaflozane; mirtazapine; bupropion; medifoxamine; tianeptine; pivagabine; venlafaxine; milnacipran; reboxetine; gepirone; duloxetine; agomelatine; desvenlafaxine

153. Psychostimulants, 154. Centrally 155. amphetamine; agents used acting dexamfetamine; metamfetamine; for ADHD sympathomimetics methylphenidate; pemoline; and fencamfamin; modafinil; nootropics armodafinil; fenozolone; atomoxetine; fenetylline; exmethylphenidate; lisdexamfetamine 156. Xanthine 157. caffeine; propentofylline derivatives 158. Other 159. meclofenoxate; pyritinol; psychostimulants piracetam; deanol; fipexide; and nootropics citicoline; oxiracetam; pirisudanol; linopirdine; nizofenone; aniracetam; acetylcarnitine; idebenone; prolintane; pipradrol; pramiracetam; adrafinil; vinpocetine; pitolisant; 160. Anti- 161. Anticholinesterases 162. tacrine; donepezil; dementia rivastigmine; galantamine drugs 163. Other anti- 164. memantine; ginkgo biloba dementia drugs Other nervous 165. Parasympa- 166. Anticholinesterases 167. neostigmine; pyridostigmine; system drugs thomimetics distigmine; ambenonium; 168. Choline 169. carbachol; bethanechol esters 170. Other 171. pilocarpine; choline parasympathomimetics alfoscerate; cevimeline 172. Drugs used 173. Drugs used 174. nicotine; varenicline in addictive in nicotine disorders dependence 175. Drugs used 176. disulfiram; calcium in alcohol carbimide; acamprosate; dependence naltrexone; baclofene 177. Drugs used 178. buprenorphine; in opioid levacetylmethadol; lofexidine; dependence 179. Antivertigo 180. Antivertigo 181. betahistine; cinnarizine; preparations preparations flunarizine; acetylleucine 182. Other 183. Other 184. tirilazad; riluzole; xaliproden; nervous nervous amifampridine; tetrabenazine; system drugs system drugs fampridine; mazindol

[0056] Preferably, the said psychotropic drug is selected in the group consisting of: dopaminergic, GABAergic, adrenergic, acetylcholinergic, serotoninergic, opioidergic, adenosinergic, ionotropic, histaminergic, IMAO, Catechol-O-methyl transferase, DOPA decarboxylase, noradrenergic and glutamatergic psychotropic effectors, as well as molecules having an effect on the hypocretin/orexin system (including hypocretin-1 and hypocretin-2).

[0057] The term "effector" herein refers to any substance activating or inhibiting, directly or indirectly, one or more neuroreceptors, as well as any substance that modifies the concentration of said neurotransmitter; therefore, an effector according to the present invention can be an agonist or an antagonist of said receptors.

[0058] It is shown in the examples below that said psychotropic drug is advantageously modafinil.

[0059] As a matter of fact, the present inventors have shown that flecainide potentiates the promnesiant and/or awakening effects of modafinil (see FIGS. 2A-B and 3), and that the modafinil/flecainide combination shows promising effects by reducing cataplectic-like events in mice. The precise mechanism of modafinil action has not been completely elucidated yet. In fact, it is known that modafinil acts on several molecular receptors, in particular on the dopamine, norepinephrine, serotonine, glutamate, GABA, orexine and histamine receptors (Ishizuka et al, 2012; Minzenberg et al, 2008). Therefore, modafinil acts as a GABAergic, dopaminergic, norepinephrinergic, serotoninergic, histaminergic, and glutamatergic effectors, and it has an effect on the hypocretin/orexin system (including hypocretin-1 and hypocretin-2).

[0060] Any compound modulating the same molecular receptors as modafinil can be advantageously associated with flecainide.

[0061] Thus, in a preferred embodiment, the psychotropic drug which is associated with flecainide acts on the very same receptors as modafinil does. The psychrotropic drug associated with flecainide is therefore preferably selected in the group consisting of: GABAergic, dopaminergic, norepinephrinergic, serotoninergic, histaminergic, and glutamatergic effectors. Also, it may have an effect on the hypocretin/orexin system (including hypocretin-1 and hypocretin-2).

[0062] According to a specific embodiment, the said psychotropic drug is a dopaminergic effector selected in the group consisting of: ADX-N05 (formely "YKP10A", having the formula: (R)-(beta-amino-benzenepropyl) carbamate mono-hydrochloride), amphetamine, loxapine, acepromazine, methylphenidate, pergolide, lisuride, bromocriptine, dopamine, ropinirole, apomorphine, aripiprazole, sulpiride, amisulpride, sultopride, tiapride, pimozide, risperidone, haloperidol, penfluridol, zuclopenthixol or bupropion.

[0063] According to another specific embodiment, the said psychotropic drug is a GABAergic effector selected in the group consisting of: tiagabine, topiramate, clorazepate, diazepam, clonazepam, oxazepam, lorazepam, bromazepam, lormetazepam, nitrazepam, clotiazepam, aiprozolam, estazolam, triazolam, loprazolam, etifoxin, meprobamate, zopiclone, zolpidem, pregabaline, gabapentine, phenobarbital, felbamate and vigabatrin.

[0064] According to another specific embodiment, the said psychotropic drug is a serotoninergic effector selected in the group consisting of: chlorpromazine, trimipramine, clozapine, olanzapine, cyamemazine, flupentixol, nefopam, fluvoxamine, clomipramine, sertraline, fluoxetine, citalopram, escitalopram, paroxetine, amitriptyline, duloxetine, venlafaxine, buspirone, carpipramine, zolmitriptan, sumatriptan, naratriptan, indoramine, ergotamine, ergotamine tartrate, pizotifene, pipamperone, methysergide, pizotyline, milnacipran, viloxazine, tianeptine, hypericum and lithium.

[0065] According to another specific embodiment, the said psychotropic drug is a histaminergic effector selected in the group consisting of: acrivastine, alimemazine, antazoline, astemizole, azatadine, azelastine, brompheniramine, buclizine, carbinoxamine, carebastine, cetirizine, chlorcyclizine, chlorpheniramine, cinnarizine, clemastine, clemizole, clocinizine, clonidine, cyclizine, cyproheptadine, descarboethoxyloratidine, dexchlorpheniramine, dimenhydrinate, dimethindene, dimethothiazine, diphenhydramine, diphenylpyraline, doxylamine, ebastine, efletirizine, epinastine, fexofenadine, hydroxyzine, ketotifen, levocabastine, loratidine, meclizine, mequitazine, methdilazine, mianserin, mizolastine, niaprazine, noberastine, norastemizole, oxatomide, oxomemazine, phenbenzamine, pheniramine, picumast, promethazine, pyrilamine, temelastine, terfenadine, trimeprazine, tripelennamine, triprolidine, ranitidine, cimetidine, famotidine, nizatidine, tiotidine, zolantidine, ciproxifan, pitolisant and ritanserine.

[0066] According to another specific embodiment, the said psychotropic drug is a hypocretin/orexin modulator selected in the group consisting of: EMPA, SB-334867, SB-674042, SB-408124, GSK1059865, almorexant, suvorexant, MK-6096, DORA-1, DORA-22, DORA-12, SB-649868, JNJ-1037049 (described in Gotter et al, 2012)).

[0067] According to another specific embodiment, the said psychotropic drug is a norepinephrinergic effector selected in the group consisting of: (R)-3-nitrobiphenyline, 2-fluoronorepinephrine, 4-NEMD, 5-fluoronorepinephrine, 6-fluoronorepinephrine, abediterol, albuterol, amibegron, amidephrine, amitraz, anisodamine, anisodine, apraclonidine, arbutamine, arformoterol, arotinolol, bambuterol, befunolol, bitolterol, brimonidine, bromoacetylalprenololmenthane, broxaterol, buphenine, cannabivarin, carbuterol, cimaterol, cirazoline, clenbuterol, denopamine, deterenol, detomidine, dexmedetomidine, dihydroergotamine, dipivefrine, dobutamine, dopexamine, ephedrine, epinephrine, esproquin, etafedrine, ethylnorepinephrine, etilefrine, fenoterol, formoterol, guanabenz, guanfacine, guanoxabenz, hexoprenaline, higenamine, indacaterol, indanidine, isoetarine, isoprenaline, isoproterenol, isoxsuprine, labetalol, levonordefrin, levosalbutamol, lofexidine, mabuterol, medetomidine, metaraminol, methoxamine, methoxyphenamine, methyldopa, midodrine, mivazerol, n-isopropyloctopamine, naphazoline, norepinephrine, octopamine, orciprenaline, oxyfedrine, oxymetazoline, phenylephrine, phenylpropanolamine, piperoxan, pirbuterol, prenalterol, procaterol, pseudoephedrine, ractopamine, reproterol, rilmenidine, rimiterol, ritodrine, romifidine, salbutamol, salmeterol, solabegron, synephrine, talipexole, terbutaline, tetrahydrozoline, tizanidine, tolonidine, tretoquinol, tulobuterol, urapidil, xamoterol, xylazine, xylometazoline, zilpaterol, and zinterol.

[0068] According to another specific embodiment, the said psychotropic drug is a glutamatergic effector selected in the group consisting of: memantine, amantadine, MK-801, ketamine, norketamine, dextromethorphan, levometorphan, dextrorphan, levorphanol, phencyclidine, PCA, CNS-1102, remacemide, pentamidine, and 9-aminoacridine (described in Traynelis et al, 2010).

[0069] Preferably, said psychotropic drug is not flupirtine.

[0070] The potentiating effects of flecainide can be achieved by administrating same to a patient, either before, at the same time, of after administration of the psychotropic drug to said patient.

[0071] Consequently, the present invention describes a method for treating a patient with psychiatric and/or neurodegenerative disorders, including the administration to said patient of a) flecainide and b) at least one psychotropic drug as mentioned above, in which said compounds a) and b) are administered simultaneously, separately or spread out over time.

[0072] Patients needing this treatment may have psychiatric, neurologic and/or neurodegenerative disorders included in the group consisting of: excessive daytime sleepiness (EDS), sleep disorders, insufficient sleep time, central sleep apnea, narcolepsy (with or without cataplexy), obstructive sleep apnea/hypopnea (SAHOS), idiopathic hypersomnia, Kleine-Levin syndrome, circadian rhythm disorders, shift work sleep disorder, jet-lag, disorders after sleep restriction or sleep deprivation (attention disorders, alertness disorders, sleepiness), restless legs syndrome (RLS) and Periodic Lim Movement Disorders (PLMD), insomnia, parasomnia, attention deficit hyperactivity disorder (ADHD), post-traumatic stress disorder (PTSD), disorders commonly associated with somnolence or sleepiness (such as Parkinson disease, multiple sclerosis, stroke, neuromuscular disorders or structural brain disorders, respiratory disorders, chronic renal failure, liver failure, rheumatologic disorders), medication-induced somnolence (due to benzodiazepines, barbiturates, sleeping pills, antidepressants, anti-psychotics . . . ), mood disorders, anxiety disorders, schizophrenia, tinnitus, depression, malaise, dementia, bipolar disorder, obesity, hyperphagia, manic episode, obsessive-compulsive disorder, senility, dependence or addiction (to games, drugs, alcohol, tobacco, etc.), fecal or urinary incontinence, premature ejaculation, breathing difficulty and fatigue, notably due to cancer, neurodegenerative disorders, menopause, traumatic brain injuries, viral infection or post-myelitis, or to fibromyalgia.

[0073] Excessive daytime sleepiness (EDS) occurs daily, recurring typically every 2 h, although this can vary widely. Sleepiness is exacerbated when the patient is physically inactive. The sleep episodes have several characteristics (Dauvilliers I. et al, 2007 and Boulos et al, 2010):

[0074] They are often irresistible, despite the individual making desperate efforts to fight the urge to sleep;

[0075] They are usually short, although their length can vary with environmental factors (eg, the duration can increase with passive activities such as watching television);

[0076] They are frequently associated with dreaming;

[0077] They typically restore normal wakefulness for up to several hours.

[0078] EDS characterizes several conditions or diseases: insufficient sleep time, central sleep apnea, narcolepsy (with or without cataplexy), obstructive sleep apnea/hypopnea (SAHOS), idiopathic hypersomnia, recurrent hypersomnia (Kleine-Levin syndrome), circadian rhythm disorders (jet lag), disorders after sleep restriction or sleep deprivation (attention disorders, alertness disorders, sleepiness), restless legs syndrome (RLS) and Periodic Lim Movement Disorders (PLMD), neurological conditions commonly associated with sleepiness (such as Parkinson disease, multiple sclerosis, stroke, neuromuscular disorders or structural brain disorders), medical conditions commonly associated with sleepiness (respiratory disorders, chronic renal failure, liver failure, rheumatologic disorders), mood disorders, anxiety disorders, schizophrenia, or medication-induced somnolence (due to benzodiazepines, barbiturates, sleeping pills, antidepressants, anti-psychotics . . . ).

[0079] Cataplexy is characterized by a sudden drop of muscle tone triggered by emotional factors, most often by positive emotions such as laughter, repartee, pleasant surprise (e.g., seeing friends in the street or scoring a goal), or by anger, but almost never by stress, fear, or physical effort. Many neurophysiological and pharmaceutical studies indicate that cataplexy shares common neurophysiological mechanisms with REM sleep atonia (Dauvilliers I. et al, 2007).

[0080] In the case of simultaneous use, the two components of the treatment are administered to the patient simultaneously. According to this embodiment of the present invention, the two components can be packaged together, in the form of a mixture, or separately, then mixed spontaneously before being administered together to the patient. Alternatively, the two components are administered simultaneously, but separately. In particular, the routes of administration of the two components may be different. The administration can also be performed at different sites. In another embodiment, the two components are administered sequentially or spaced apart over time, for example in the same day or at an interval ranging from several minutes to several days.

[0081] Since flecainide potentiates the effects of psychotropic drugs, it can advantageously be used for reducing the doses of said psychotropic drug, thereby limiting the adverse effects of said psychotropic drug, and/or reducing the risks of failure and withdrawal.

[0082] The effective equivalent dose of a psychotropic drug, i.e., the psychotropic drug dose that, when administered in combination with flecainide, induces a physiological effect or a pharmacological signature similar or identical to that of the psychotropic drug alone administered at the active pharmacological dose, can be determined by the methods disclosed in WO2010/029131 and US 2011/172188.

[0083] According to another aspect, the present invention pertains to a composition, especially a pharmaceutical composition, comprising flecainide and at least one psychotropic drug. This composition is preferably formulated for patients with psychiatric and/or neurodegenerative disorders, as disclosed above. In addition to flecainide and to said psychotropic drug, the composition can comprise any pharmaceutical vehicle, stabilizer, adjuvant and the like as frequently used in the art.

[0084] Examples of pharmaceutically acceptable vehicles include, but are not limited to: water; aqueous vehicles such as, but not limited to, sodium chloride solution, Ringer's solution, dextrose solution, dextrose and sodium chloride solution, and lactated Ringer's solution; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and polypropylene glycol; and nonaqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

[0085] According to a preferred embodiment, this composition is formulated for oral administration (including buccal cavity or sublingually). Other interesting formulations include formulations for intraperitoneal (i.p.), intravenous (i.v.), subcutaneous (s.c.), intramuscular (i.m.), transcutaneous, transdermal, intrathecal and intracranial administrations. Still other formulations include epidural, submucosal, intranasal, ocular cul-de-sac and rectal routes of administration, as well as administration by pulmonary inhalation.

[0086] A variety of administration means, including but not limited to capsules, tablets, syrups, creams and ointments, suppositories, patches or any reservoir capable of containing and dispensing flecainide and the psychotropic drug, can be used for formulating the above-described compositions.

[0087] In the compositions according to the invention, the psychotropic drug is as described above.

[0088] In a preferred embodiment, said psychotropic drug is used for treating narcolepsy and is therefore selected in the group consisting of: caffeine, mazindol, sodium oxybate, pitolisant, amphetamine, methylphenidate, (R)-(beta-amino-benzenepropyl) carbamate mono-hydrochloride, modafinil and armodafinil.

[0089] In a preferred embodiment, the composition of the invention contains between 1 to 1000 mg, preferably 5 to 800 mg of the psychotropic drug, depending of its nature. A preferred posology would be to administer to the patient between 1 to 1000 mg/day, more preferably between 5 to 800 mg/day of the psychotropic drug.

[0090] According to another preferred embodiment, the composition of the invention contains between 1 to 200, preferably 1 to 100 mg of flecainide. A preferred posology would be to administer to the patient between 1 to 200, preferably 1 to 100 mg/day of flecainide.

[0091] More preferably, said flecainide is the R enantiomer disclosed on FIG. 5A.

[0092] In a more preferred embodiment, flecainide is associated with the psychotropic drug modafinil.

[0093] By "modafinil" is herein meant the 2-[(diphenylmethyl) sulfinyl] acetamide (Provigil, see FIG. 5C), as well as its precursors or prodrugs such as adrafinil (Dubey et al, 2009) which can be metabolized in the human body and its active derivatives. More precisely, the term "Modafinil" herein designates any form of modafinil (racemate, R-modafinil, S-modafinil, etc.), as well as its precursors which can be metabolized in the human body and its derivatives. FIGS. 5C-D show the formulas of R-Modafinil (FIG. 5C) and S-Modafinil (FIG. 5D).

[0094] Modafinil is an analeptic drug prescribed essentially for the treatment of narcolepsy, shift work sleep disorder, and excessive daytime sleepiness associated with obstructive sleep apnea. Besides these wake-promoting properties, modafinil also improves working memory and episodic memory, and other processes dependent on prefrontal cortex and cognitive control (Minzenberg M J et al, 2008).

[0095] The present inventors have shown that, surprisingly, flecainide strongly potentiates in vivo the waking effects of Modafinil, whereas it has no effect on wake duration on its own (example 2). Moreover, flecainide strongly potentiates in vivo the cognitive activity of Modafinil, whereas it has no promnesiant effect on its own (example 3). This synergistic activity could be explained by the fact that flecainide strongly extends the duration of Modafinil treatment (example 4). On the other hand, the present inventors herein describes that the flecainide/modafinil combination has a synergistic effect on cataplectic-like phenotype in narcoleptic mice (example 5) and is all the more surprising than either flecainide or modafinil has no effect on this phenotype (FIG. 6B). In a preferred embodiment, the present invention thus pertains to flecainide, for use for potentiating the promnesiant and/or awakening effects of modafinil, and/or for improving its safety, and/or for increasing the duration of action of modafinil in patients in need thereof, especially in patients suffering from: excessive daytime sleepiness (EDS), sleep disorders, insufficient sleep time, central sleep apnea, narcolepsy (with or without cataplexy), obstructive sleep apnea/hypopnea (SAHOS), idiopathic hypersomnia, Kleine-Levin syndrome, circadian rhythm disorders, shift work sleep disorder, jet-lag, disorders after sleep restriction or sleep deprivation (attention disorders, alertness disorders, sleepiness), restless legs syndrome (RLS) and Periodic Lim Movement Disorders (PLMD), insomnia, parasomnia, attention deficit hyperactivity disorder (ADHD), post-traumatic stress disorder (PTSD), disorders commonly associated with somnolence or sleepiness (such as Parkinson disease, multiple sclerosis, stroke, neuromuscular disorders or structural brain disorders, respiratory disorders, chronic renal failure, liver failure, rheumatologic disorders), medication-induced somnolence (due to benzodiazepines, barbiturates, sleeping pills, antidepressants, anti-psychotics . . . ), mood disorders, anxiety disorders, schizophrenia, tinnitus, depression, malaise, dementia, bipolar disorder, obesity, hyperphagia, manic episode, obsessive-compulsive disorder, senility, dependence or addiction (to games, drugs, alcohol, tobacco, etc.), fecal or urinary incontinence, premature ejaculation, breathing difficulty and fatigue, notably due to cancer, neurodegenerative disorders, menopause, traumatic brain injuries, viral infection or post-myelitis, or to fibromyalgia, which have been proposed to be treated by modafinil.

[0096] In a more preferred embodiment, the present invention specifically pertains to flecainide, for use for potentiating the awakening effects of modafinil, and/or for improving its safety, and/or for increasing the duration of action of modafinil in patients suffering from: excessive daytime sleepiness (EDS), sleep disorders, insufficient sleep time, central sleep apnea, narcolepsy (with or without cataplexy), obstructive sleep apnea/hypopnea (SAHOS), idiopathic hypersomnia, Kleine-Levin syndrome, circadian rhythm disorders, shift work sleep disorder, jet-lag, disorders after sleep restriction or sleep deprivation (attention disorders, alertness disorders, sleepiness), restless legs syndrome (RLS) and Periodic Lim Movement Disorders (PLMD), insomnia, parasomnia, attention deficit hyperactivity disorder (ADHD), post-traumatic stress disorder (PTSD), disorders commonly associated with somnolence or sleepiness (such as Parkinson disease, multiple sclerosis, stroke, neuromuscular disorders or structural brain disorders, respiratory disorders, chronic renal failure, liver failure, rheumatologic disorders), medication-induced somnolence (due to benzodiazepines, barbiturates, sleeping pills, antidepressants, anti-psychotics . . . ), mood disorders, anxiety disorders, schizophrenia, tinnitus, depression, malaise, dementia, bipolar disorder, obesity, hyperphagia, manic episode, obsessive-compulsive disorder, senility, dependence or addiction (to games, drugs, alcohol, tobacco, etc.), fecal or urinary incontinence, premature ejaculation, breathing difficulty and fatigue, notably due to cancer, neurodegenerative disorders, menopause, traumatic brain injuries, viral infection or post-myelitis, or to fibromyalgia, for which modafinil has been proposed or authorized.

[0097] In a preferred embodiment, the present invention specifically pertains to flecainide, for use for potentiating the awakening effects of modafinil, and/or for improving its safety, and/or for increasing the duration of action of modafinil in patients suffering from excessive daytime sleepiness (EDS).

[0098] In another preferred embodiment, the present invention relates to flecainide, for use for potentiating the awakening effects of modafinil, and/or for improving its safety, and/or for increasing the duration of action of modafinil in patients suffering from conditions or diseases involving EDS, that are for example: insufficient sleep time, central sleep apnea, narcolepsy (with or without cataplexy), obstructive sleep apnea/hypopnea (SAHOS), idiopathic hypersomnia, recurrent hypersomnia (Kleine-Levin syndrome), circadian rhythm disorders (jet lag), disorders after sleep restriction or sleep deprivation (attention disorders, alertness disorders and sleepiness), restless legs syndrome (RLS) and Periodic Lim Movement Disorders (PLMD), neurological conditions commonly associated with sleepiness (such as Parkinson disease, multiple sclerosis, stroke, neuromuscular disorders or structural brain disorders), medical conditions commonly associated with sleepiness (respiratory disorders, chronic renal failure, liver failure, rheumatologic disorders), mood disorders, anxiety disorders, schizophrenia, or medication-induced somnolence (due to benzodiazepines, barbiturates, sleeping pills, antidepressants, anti-psychotics . . . ).

[0099] In another preferred embodiment, the present invention relates to a modafinil/flecainide combination product, for use for treating cataplexy in narcoleptic patients.

[0100] It is to be noted that the potentiation of the effects of modafinil by flecainide enables a reduction of the dose of modafinil, and hence a reduction of its side-effects. As a consequence, some applications of modafinil, for which this drug was not approved because of its side-effects and possible risks associated thereto, can now be envisioned, such as its use as a performance-enhancing and/or brain-boosting agent. According to a particular embodiment, the present invention thus pertains to a performance-enhancing product comprising flecainide and modafinil.

[0101] In another preferred embodiment, the present invention specifically pertains to the use of flecainide and modafinil for enhancing the memory of healthy subjects and/or to maintain them awake for long-lasting periods of time and/or to treat cataplexy in narcoleptic patients. These subjects can be for example individuals that need to memorize a lot of information and/or to remain awake for long lasting periods. In a preferred embodiment, said subjects are humans (e.g., security agents, students, etc.).

[0102] In a particular embodiment, the present invention also relates to a composition comprising flecainide and modafinil, which can advantageously be used for treating diseases and conditions including but not limited to excessive daytime sleepiness (EDS), narcolepsy (with or without cataplexy), obstructive sleep apnea/hypopnea (SAHOS), shift work sleep disorder, disorders after sleep restriction or sleep deprivation (attention disorders, alertness disorders, sleepiness), restless leg syndrome, hypersomnia, idiopathic hypersomnia and fatigue, notably due to cancer, jet-lag, neurodegenerative disorders, menopause, traumatic brain injuries, viral infection or post-myelitis, or to fibromyalgia. In particular, this composition can be used for treating cataplexy in narcoleptic patients.

[0103] This composition can also be used for enhancing the memory of healthy subjects and/or for maintaining them awake for long-lasting periods of time. Typical periods of time are for example 6 hours, preferably 12 hours.

[0104] The present invention moreover relates specifically to the use of flecainide and modafinil in the preparation of a medicament that is intended to be used for treating diseases and conditions such as excessive daytime sleepiness (EDS), narcolepsy (with or without cataplexy), obstructive sleep apnea/hypopnea (SAHOS), shift work sleep disorder, restless leg syndrome, hypersomnia, idiopathic hypersomnia and fatigue, notably due to cancer, neurodegenerative disorders, menopause, traumatic brain injuries, viral infection or post-myelitis, or to fibromyalgia.

[0105] In a preferred embodiment, the present invention relates to the use of flecainide and modafinil in the preparation of a medicament that is intended to be used for treating cataplexy in narcoleptic patients.

[0106] In addition to modafinil and flecainide, the composition/medicament of the invention can comprise other agents such as vitamin C, vitamin B6, magnesium, L-arginine, L-glutamine, L-citrulline, taurine, caffeine, etc. According to a particular embodiment, this product can be sold over-the-counter. It can be formulated, for example, as an OTC medicine or as a food supplement.

[0107] In a preferred embodiment, the composition of the invention contains between 1 to 1000 mg, preferably between 5 to 800 mg, and more preferably between 5 to 600 mg of the modafinil. According to another preferred embodiment, the composition of the invention is formulated so that 5 to 800, preferably 5 to 600 mg/day of modafinil are administered to a patient in need thereof, in one, two or more takings.

[0108] According to another preferred embodiment, the composition of the invention contains between 1 to 200, preferably 1 to 100 mg of flecainide. According to another preferred embodiment, the composition of the invention is formulated so that 1 to 200, preferably 1 to 100 mg/day of flecainide are administered to a patient in need thereof, in one, two or more takings. In a more preferred embodiment, said flecainide is the R enantiomer disclosed on FIG. 5A.

[0109] In a final aspect, the present invention relates to a combination product comprising flecainide and modafinil, for simultaneous, separated or staggered use for preventing and/or treating excessive daytime sleepiness (EDS), narcolepsy (with or without cataplexy), obstructive sleep apnea/hypopnea (SAHOS), shift work sleep disorder, restless leg syndrome, hypersomnia, idiopathic hypersomnia and fatigue, notably due to cancer, jet-lag, neurodegenerative disorders, menopause, traumatic brain injuries, viral infection or post-myelitis, or to fibromyalgia. This combination product is preferably used for preventing and/or treating cataplexy in narcoleptic patients.

[0110] Other characteristics of the invention will also become apparent in the course of the description which follows of the biological assays which have been performed in the framework of the invention and which provide it with the required experimental support, without limiting its scope.

LEGENDS TO THE FIGURES

[0111] FIGS. 1A-B: Inhibition of the human connexins functionality by flecainide. Rin-Cx26 cells, Rin-Cx30 cells, Rin-Cx32 cells, Rin-CX40 cells and Rin-Cx43 cells are cultured in the presence of flecainide (280 .mu.M), mefloquine (10 .mu.M) and MFA (100 .mu.M) for 4 hours. The transfer of fluorochrome by gap junctions (composed of connexins) is evaluated by flow cytometry (1A and 1B). Viability of the cells treated with flecainide is shown on FIG. 1B.

[0112] FIGS. 2A-B: Efficiency of flecainide for potentiating the awakening effect of modafinil. Mice (n=8 per batch) were orally treated by either modafinil (32 mg/kg) or modafinil (32 mg/kg) and flecainide (1 mg/kg) (FIG. 2A) or flecainide alone (1 mg/kg) (FIG. 2B) and replaced in their home cage. The wake duration was measured using polygraphic analyses.

[0113] FIG. 3: Efficacy of flecainide for potentiating the promnesiant effect of modafinil. Mice (n=6 to 23 per batch) are tested in the T-maze. They were intraperitoneally treated by either modafinil (64 mg/kg or 128 mg/kg) or modafinil (64 mg/kg) and flecainide (1 mg/kg) or flecainide alone (1 mg/kg). The graphic represents the percentage of alternation after 6 trials, 50% corresponding to a random alternation.

[0114] FIG. 4: Efficacy of flecainide for potentiating the locomotor effect of modafinil. Mice (n=8 per batch) were orally treated by either modafinil (64 mg/kg) or modafinil (64 mg/kg) and flecainide (1 mg/kg) or flecainide alone (1 mg/kg) and replaced in their home cage. The locomotor activity was measured using videotracking device.

[0115] FIGS. 5A-D: Molecular structure of FIG. 5A. R-flecainide; FIG. 5B. S-flecainide; FIG. 5C. R-Modafinil, FIG. 5D. S-Modafinil.

[0116] FIGS. 6A-B: Number of episodes of OREM/DREM phases in narcoleptic mice (Ox-/-) treated by modafinil/flecainide (FIG. 6A) or flecainide alone (FIG. 6B). (FIG. 6A). Oral treatment of Ox-/- male mice with modafinil 64 mg/kg with flecainide 1 mg/kg was compared to Modafinil 64 mg/kg and vehicle. **: p<0.01; ***: p<0.005, Two-Way ANOVA. (FIG. 6B) Oral treatment of Ox-/- male mice with flecainide 1 mg/kg was compared to vehicle.

[0117] FIG. 7: Number of episodes of OREM/DREM phases in narcoleptic mice (Ox-/-) treated by the combination between modafinil and one of the two enantiomers of flecainide (R-flecainide and S-flecainide). Oral treatment with modafinil 64 mg/kg with R-flecainide 1 mg/kg or S-flecainide 1 mg/kg was compared to vehicle.

EXAMPLES

Example 1: Effect of Flecainide on Gap Junctions

1.1. Materials and Methods

Cell Culture

[0118] The rat insulinoma RIN cell line, deficient in GJIC (del Corsso et al, 2006), was grown in OptiMem medium, supplemented with 10% fetal calf serum. GJB6 (Cx30), GJB1 (Cx32), GJB2 (Cx26), GJA5 (Cx40) and GJA1 (Cx43) open reading frames were amplified from human cDNA. The open reading frames were cloned in pcDNA3.1/V5-His-TOPO (Invitrogen). Cells were transfected using Lipofectamine and further selected using geneticin.

Dye Transfer Experiments

[0119] Cells were seeded and loaded with two fluorochromes, calcein acetoxymethyl ester, a gap junction permeable dye, and Vybrant Dil, a membrane lipophilic dye. The next day, cells were dissociated and incubated for four hours in presence of previously seeded non-loaded cells and in the presence of flecainide racemate 70, 140 or 280 .mu.M, mefloquine 10 .mu.M or meclofenamic acid (MFA) 100 .mu.M. Flow cytometry was conducted on a FACScan. Inhibition was quantified as the relative number of receiver cells that gained fluorescence to the total number of receiver cells (non GJ-mediated dye transfer was then substracted to these ratio based on connexin non-expressing RIN cells, defined at background dye transfer ratio). This ratio of cellular coupling was then normalized, after each treatment, on the vehicle one.

Toxicity Analysis

[0120] Twenty thousand RIN were seeded in 100 .mu.l of culture medium in 96-wells plates. After 48 h culture, cells were treated for 4 hours with previously identified chemical compounds at several concentrations. Cells were rinsed in PBS and grown 24 h in fresh medium. Cell viability was measured by using WST-1 (Roche).

1.2. Experimental Results

[0121] Cellular models were validated by using two classical inhibitors described in litterature, meclofenamic acid (MFA) (Dhein, 2004) (100 .mu.M) and mefloquine (Cruikshank et al, 2004) (10 .mu.M). Results are shown on FIG. 1A. Flecainide is as efficient in blocking connexin as the other anti-connexin agents.

[0122] Cell viability tests (using WST-1, dotted curve on FIG. 1B) after one day of treatment, indicate that flecainide has no cell toxicity at the dose inhibiting cerebral connexins.

[0123] In addition, flecainide inhibits all the tested isoforms of cerebral connexin using dye-transfer cell-parachute assay (Cx30, Cx32, Cx26, Cx40, Cx43) (it is estimated that a more than a significant 10% reduction in gap junction cellular is considered as physiologically relevant). In addition, higher inhibition levels are reached for glial connexins Cx26, Cx30 and Cx43.

Example 2: Flecainide Potentiates the Waking Effects of Modafinil

[0124] Preclinical and clinical data indicated that modafinil modifies sleep-cycle rhythm and promotes wake phases (Lin et al, 2008). Here we tested in rodents whether such activity was potentiated by flecainide after oral challenge with modafinil, using polysomnographic analysis on implanted mice. Using a sub-efficient dosage of modafinil (32 mg/kg), the inventors demonstrated a new feature of the combination of modafinil and flecainide since it significantly increases the total duration of wake episodes.

2.1. Materials and Methods

[0125] Wild-type C57bl/6 male mice (n=9/groups) were implanted with EEG/EMG/EOG electrodes for polysomnographic analyses. After a two-week recovery period, mice were orally treated with vehicle, Modafinil 32 mg/kg and Modafinil 32 mg/kg+flecainide racemate 1 mg/kg and wake periods were quantified using Spike2 scripts. Here the inventors represented the duration of wake during the first three hours (after a one-hour period post-administration). **: p<0.01 in a One-Way ANOVA analysis.

2.2. Results

[0126] Modafinil is a molecule that promotes wakefulness in humans and mice, increasing in mice their activity in a dose-dependent manner (Simon et al, 1994). The activity of mice treated with modafinil at 32 mg/kg was compared with that of mice treated with the combination modafinil 32 mg/kg+flecainide 1 mg/kg or vehicle.

[0127] FIG. 2A shows that flecainide significantly increases the waking effects of modafinil. FIG. 2B shows that this effect is not mediated by flecainide alone.

[0128] Thus, flecainide significantly potentiates modafinil waking activity in wild type mice, while being devoid of own effect on wake duration.

Example 3: Flecainide Significantly Enhances Modafinil Cognitive Activity

[0129] Modafinil induces a cognitive enhancing effect (Beracochea et al, 2003), such property can be assessed using the alternating sequential test, a widely used apparatus to assess spatial working memory in mice (Beracochea & Jaffard, 1987). Spontaneous alternation is the innate tendency of rodents to alternate their choices to enter into the compartments of arrival of a T-maze device, over successive trials. To alternate during a given trial N, the animal must remember the choice made selectively in test N-1, and the response in alternating is performance measure. Acute administration of modafinil before entering the maze, can improve the performance of mice in this test (Beracochea et al, 2001). The inventors' results showed that flecainide significantly potentiates the promnesiant effect of a subefficient dose of modafinil, while flecainide alone is devoid of any own promnesiant effect.

3.1. Materials and Methods

[0130] The alternating sequential test is widely used to assess spatial working memory in mice (Beracochea & Jaffard, 1987). Spontaneous alternation is the innate tendency of rodents to alternate their choices to entry into the compartments of arrival of a T-maze device, over successive trials. To alternate during a given trial N, the animal must remember the choice made selectively in test N-1, so the decline in alternating will reflect the phenomenon of oblivion. The response in alternating is performance measure. Sequential alternating assesses more specifically the sensitivity to interference, a major factor in oblivion.

[0131] The experiment takes place in a T-maze (50 cm.times.10 cm.times.25 cm). All the subjects were given 7 successive trials separated by a 120-s intertrial interval. To begin a trial, the mouse was placed in the start box for 120 s before the door to the stem was opened. When the subject entered one of the goal arms, the door to that arm was closed. The chosen arm and the time that elapsed between opening the door and the arrival to the end of the chosen arm (task achievement time) were registered. Following a 30-s confinement period (fixed and invariant) in the chosen arm, the animal was removed and placed in the start box for a new trial. Between each test, the unit is cleaned with a cloth soaked in water and alcohol to avoid olfactory detection. The index memory is represented by the average of alternating percentage (number of alternation choices/total number of tests X 100). (n=6 to 23 for each group). Mice were intraperitoneally treated by either modafinil (64 mg/kg or 128 mg/kg) or modafinil (64 mg/kg) and flecainide racemate (1 mg/kg) or flecainide racemate alone (1 mg/kg) or vehicle.

[0132] # p<0.05 in one sample t-test vs random 50% alternance; * p<0.05 One way ANOVA followed by Tukey's multiple comparison vs modafinil group.

3.2. Results

[0133] The T-maze is a device for assessing working memory in mice. Acute administration of modafinil before entering the maze, can improve the performance of mice in this test (Beracochea et al, 2001).

[0134] The validity of the test was performed by comparing the responses of mice intraperitoneally treated with an effective dose of modafinil alone (128 mg/kg), a dose of flecainide alone (1 mg/kg) and a sub-effective dose of modafinil (64 mg/kg) with or without flecainide alone (1 mg/kg). The results are shown in FIG. 3.

[0135] These results show that flecainide significantly potentiates modafinil promnesiant activity, while flecainide alone shows no own cognitive effect.

Example 4: Flecainide Significantly Prolongs Modafinil Activity

[0136] Modafinil is a molecule that promotes wakefulness in humans and mice, increasing in mice their activity in a dose-dependent manner (Simon et al, 1994). The inventors' results showed that flecainide significantly potentiates the locomotor effect of a subefficient dose of modafinil, while flecainide alone is devoid of any own locomotor effect in rodents.

4.1. Materials and Methods

[0137] Mice (n=8 per batch) were orally treated by either modafinil (64 mg/kg) or modafinil (64 mg/kg) and flecainide racemate (1 mg/kg) or flecainide racemate alone (1 mg/kg) or vehicle and replaced in their home cage. Locomotor activity is evaluated by video tracking. Videos have been analyzed using Ethovision XT software (Noldus.RTM.). *: p<0.01 in a Two-Way ANOVA analysis

4.2. Results

[0138] The activity of mice treated with modafinil at 64 mg/kg was compared with that of mice treated with the combination modafinil 64 mg/kg+flecainide 1 mg/kg. FIG. 4 shows that flecainide significantly increases the duration of effect of modafinil on the activity of mice.

[0139] To conclude, the above results show that Flecainide significantly inhibits the functionality of gap junctions, without inducing cellular toxicity. In addition, this compound potentiates the efficacy and duration of effect of modafinil, notably in its promnesiant and awakening side.

Example 5: Modafinil/Flecainide Combination has a Surprising Efficient Profile on DREM Cataplectic-Like Phenotype in Narcoleptic Mice

5.1. Material and Methods

Animals

[0140] Prepro-orexin knockout (KO) mice were offspring of the mouse strain generated by Chemelli et al.

[1999] and kept on C57BL/6J genomic background. After backcrossing male orexin-/- mice and female wild-type (WT) mice for nine generations, the obtained orexin+/- mice were crossed to produce heterozygote and homozygote WT and KO littermates. To determine their genotypes with respect to orexin gene, tail biopsies were performed at the age of 4 weeks for DNA detection using PCR.

Surgery

[0141] At the age of 12 weeks and with a body weight of 30.+-.2 g, mice used for EEG and sleep-wake studies were chronically implanted, under deep gas anesthesia using isoflurane (2%, 200 ml/min) and a TEM anesthesia system (Bordeaux, France), with six cortical electrodes (gold-plated tinned copper wire, O=0.4 mm, Filotex, Draveil, France) and three muscle electrodes (fluorocarbon-coated gold-plated stainless steel wire, O=0.03 mm, Cooner Wire Chatworth, Calif., U.S.A.) to record the electroencephalogram (EEG) and electromyogram (EMG) and to monitor the sleep-wake cycle. All electrodes were previously soldered to a multi-channel electrical connector and each was separately insulated with a covering of heat-shrinkable polyolefin/polyester tubing. Cortical electrodes were inserted into the dura through 3 pairs of holes made in the skull, located respectively in the frontal (1 mm lateral and anterior to the bregma), parietal (1 mm lateral to the midline at the midpoint between the bregma and lambda), and occipital (2 mm lateral to the midline and 1 mm anterior to the lambda) cortex. Muscle electrodes were inserted into the neck muscles. Finally, the electrode assembly was anchored and fixed to the skull with Super-Bond (Sun Medical Co., Shiga, Japan) and dental cement. This implantation allows stable and long-lasting polygraphic recordings [Parmentier et al, 2002].

Polygraphic Recording in the Mouse and Data Acquisition and Analysis

[0142] After surgery, the animals were housed individually, placed in an insulated sound-proof recording room maintained at an ambient temperature of 23.+-.1.degree. C. and on a 12 h light/dark cycle (lights-on at 7 a.m.). After a 7-day recovery period, mice were habituated to the recording cable for 7 days before polygraphic recordings were started. Direct REM sleep onset (DREMs) episodes, also called narcoleptic episodes or sleep onset REM periods by some authors [Chemelli et al, 1999; Mignot et al, 2005; Fujiki et al, 2006], were defined as the occurrence of REM sleep directly from wake, namely a REM episode that follows directly a wake episode lasting more than 60 s without being preceded by any cortical slow activity of more that 5 s during the 60 s.

Drug Administration and Experimental Procedures in the Mouse

[0143] After recovery from the surgery and habituation to the recording cables, each mouse was subjected to a recording session of two continuous days, beginning at 7 a.m. Administrations were performed at 6:45 p.m. just before lights-off (7:00 p.m.), since orexin-/- mice display narcoleptic attacks only during lights-off phase [Chemelli et al, 1999]. The order of administration was randomized. Polygraphic recordings were made immediately after administration and were maintained during the whole lights-off period (12 h). Two administrations were separated by a period of 7 days (washout). Mice (n=8 per batch) were orally treated by either modafinil (64 mg/kg) or modafinil (64 mg/kg) and flecainide racemate (1 mg/kg) or flecainide racemate alone (1 mg/kg) or vehicle.

5.2. Results

[0144] Orexins (also known as hypocretins) are two hypothalamic neuropetides identified in 1998 [Sakurai et al, 1998; De Lecea L. et al, 1998]. Neurons containing orexins have been identified in the hypothalamic dorsolateral and peri-fornical areas, these neurons play a key role in behavioral arousal. A large body of evidence indicates that an orexin deficiency is responsible for the pathogenesis of human and animal narcolepsy [Lin et al, 1999; Chemelli et al, 1999]. It has been recently shown that the most major phenotypes of orexin KO mice are a behavior/motor deficit during waking and the occurrence, during the dark phase, of episodes of sleep onset REM (DREM, as known as SOREM)--defined on EEG, EMG and video recordings as sudden onset of paradoxical sleep directly from wakefulness [Anaclet et al, 2009]. Thus SOREM/DREM constitutes a main phenotype of murine narcolepsy which is frequently seen in narcoleptic patients [Lin et al, 20011]. Using this model, it was shown that modafinil allows DREM episodes to persist [Lin et al, 2008], a situation similar to that in the clinic in which modafinil improves excessive daytime sleepiness without clear effect in cataplexy.

[0145] Moreover, as disclosed on FIG. 6B, flecainide racemate (alone), at 1 mg/kg dose, has no effect on DREM cataplectic-like phenotype in narcoleptic Ox-/- mice.

[0146] However, and importantly, the results disclosed on FIG. 6A show that modafinil/flecainide combination decreases the occurrence of DREM episode.

[0147] Hence, flecainide and modafinil do not have any significant effect on a DREM cataplectic-like phenotype when used alone, whereas their combination importantly decreases the DREM cataplectic-like phenotype.

[0148] These results highlight the synergy existing between flecainide and modafinil, said synergy being due to the potentiation of the modafinil efficiency by flecainide, since no effect is seen with either modafinil or flecainide alone in narcoleptic mice.

Example 6: Modafinil/R-Flecainide is Surprisingly More Efficient than Modafinil/S-Flecainide on DREM Cataplectic-Like Phenotype in Narcoleptic Mice

[0149] The same materials and methods than in example 5 were used, except that the flecainide racemate has been replaced by the R-flecainide enantiomer.

[0150] As disclosed on FIG. 7, R-flecainide enantiomer combined with modafinil is more efficient on DREM cataplectic-like phenotype in narcoleptic Ox-/- mice than the S-flecainide enantiomer combined with modafinil.

REFERENCES

[0151] S. Alessi-Severini, F. Jamali, F. M. Pasutto, R. T. Coutts, S. Gulamhusein, High-performance liquid chromatographic determination of the enantiomers of flecainide in human plasma and urine, J Pharm Sci 79 (1990) 257-260.

[0152] S. Alessi-Severini, D. F. LeGatt, F. M. Pasutto, F. Jamali, R. T. Coutts, HPLC analysis of flecainide enantiomers in plasma: comparison with fluorescence polarization immunoassay, Clin Chem 37 (1991) 111-112.

[0153] Anaclet C, Parmentier R, Ouk K, Guidon G, Buda C, Sastre J P, Akaoka H, Sergeeva O A, Yanagisawa M, Ohtsu H, Franco P, Haas H L, Lin J S (2009) Orexin/hypocretin and histamine: distinct roles in the control of wakefulness demonstrated using knock-out mouse models. The Journal of neuroscience: the official journal of the Society for Neuroscience 29: 14423-14438

[0154] E. H. Banitt, J. R. Schmid, R. A. Newmark, Resolution of flecainide acetate, N-(2-piperidylmethyl)-2,5-bis(2,2,2-trifluoroethoxy)benzam ide acetate, and antiarrhythmic properties of the enantiomers, J Med Chem 29 (1986) 299-302.

[0155] Beracochea D, Cagnard B, Celerier A, le Merrer J, Peres M, Pierard C (2001) First evidence of a delay-dependent working memory-enhancing effect of modafinil in mice. Neuroreport 12: 375-378

[0156] Beracochea D, Celerier A, Peres M, Pierard C (2003) Enhancement of learning processes following an acute modafinil injection in mice. Pharmacology, biochemistry, and behavior 76: 473-479

[0157] Beracochea D J, Jaffard R (1987) Impairment of spontaneous alternation behavior in sequential test procedures following mammillary body lesions in mice: evidence for time-dependent interference-related memory deficits. Behav Neurosci 101: 187-197

[0158] M. I. Boulos M. I., B. J. Murray, Current evaluation and management of excessive daytime sleepiness, Can J Neurol Sci 37 (2010) 167-176

[0159] Chemelli R M, Willie J T, Sinton C M, Elmquist J K, Scammell T, Lee C, Richardson J A, Williams S C, Xiong Y, Kisanuki Y, Fitch T E, Nakazato M, Hammer R E, Saper C B, Yanagisawa M (1999) Narcolepsy in orexin knockout mice: molecular genetics of sleep regulation. Cell 98: 437-451

[0160] Cruikshank S J, Hopperstad M, Younger M, Connors B W, Spray D C, Srinivas M (2004) Potent block of Cx36 and Cx50 gap junction channels by mefloquine. Proc Natl Acad Sci USA 101: 12364-12369

[0161] Daleau P (1998) Effects of antiarrhythmic agents on junctional resistance of guinea pig ventricular cell pairs. The Journal of pharmacology and experimental therapeutics 284: 1174-1179

[0162] Dauvilliers Y., I. Arnulf, E. Mignot, Narcolepsy with cataplexy, Lancet 369 (2007) 499-511.

[0163] Davidson J O, Green C R, Nicholson L F, Bennet L, Gunn A J (2013) Connexin hemichannel blockade is neuroprotective after, but not during, global cerebral ischemia in near-term fetal sheep. Experimental neurology

[0164] de Lecea L, Kilduff T S, Peyron C, Gao X, Foye P E, Danielson P E, Fukuhara C, Battenberg E L, Gautvik V T, Bartlett F S, 2nd, Frankel W N, van den Pol A N, Bloom F E, Gautvik K M, Sutcliffe J G (1998) The hypocretins: hypothalamus-specific peptides with neuroexcitatory activity. Proc Natl Acad Sci USA 95: 322-327

[0165] del Corsso C, Srinivas M, Urban-Maldonado M, Moreno A P, Fort A G, Fishman G I, Spray D C (2006) Transfection of mammalian cells with connexins and measurement of voltage sensitivity of their gap junctions. Nat Protoc 1: 1799-1809

[0166] Dhein S (2004) Pharmacology of gap junctions in the cardiovascular system. Cardiovasc Res 62: 287-298

[0167] Dubey S, Ahi S, Reddy I M, Kaur T, Beotra A, Jain S (2009) A novel study of screening and confirmation of modafinil, adrafinil and their metabolite modafinilic acid under EI-GC-MS and ESI-LC-MS-MS ionization. Indian journal of pharmacology 41: 278-283

[0168] Durham P L, Garrett F G (2009) Neurological mechanisms of migraine: potential of the gap-junction modulator tonabersat in prevention of migraine. Cephalalgia 29 Suppl 2: 1-6

[0169] N. Fujiki, Y. Yoshida, S. Zhang, T. Sakurai, M. Yanagisawa, S. Nishino, Sex difference in body weight gain and leptin signaling in hypocretin/orexin deficient mouse models, Peptides 27 (2006) 2326-2331.

[0170] Gotter A L, Webber A L, Coleman P J, Renger J J, Winrow C J (2012) International Union of Basic and Clinical Pharmacology. LXXXVI. Orexin receptor function, nomenclature and pharmacology. Pharmacological reviews 64: 389-420

[0171] A. S. Gross, G. Mikus, C. Fischer, R. Hertrampf, U. Gundert-Remy, M. Eichelbaum, Stereoselective disposition of flecainide in relation to the sparteine/debrisoquine metaboliser phenotype, Br J Clin Pharmacol 28 (1989) 555-566.

[0172] K. Hanada, S. Akimoto, K. Mitsui, M. Hashiguchi, H. Ogata, Quantitative determination of disopyramide, verapamil and flecainide enantiomers in rat plasma and tissues by high-performance liquid chromatography, J Chromatogr B Biomed Sci Appl 710 (1998) 129-135.

[0173] Harks E G, de Roos A D, Peters P H, de Haan L H, Brouwer A, Ypey D L, van Zoelen E J, Theuvenet A P (2001) Fenamates: a novel class of reversible gap junction blockers. The Journal of pharmacology and experimental therapeutics 298: 1033-1041

[0174] Ishizuka T, Murotani T, Yamatodani A (2012) Action of modafinil through histaminergic and orexinergic neurons. Vitamins and hormones 89: 259-278

[0175] Juszczak G R, Swiergiel A H (2009) Properties of gap junction blockers and their behavioural, cognitive and electrophysiological effects: animal and human studies. Prog Neuropsychopharmacol Biol Psychiatry 33: 181-198

[0176] H. K. Kroemer, J. Turgeon, R. A. Parker, D. M. Roden, Flecainide enantiomers: disposition in human subjects and electrophysiologic actions in vitro, Clin Pharmacol Ther 46 (1989) 584-590

[0177] Laird D W (2006) Life cycle of connexins in health and disease. Biochem J 394: 527-543

[0178] A. H. L. Lie, R. M. Stuurman, F. N. Ijdenberg, J. H. Kingma, D. K. Meijer, High-performance liquid chromatographic assay of flecainide and its enantiomers in serum, Ther Drug Monit 11 (1989) 708-711.

[0179] A. H. L. Lie, J. van den Akker, A. den Hertog, D. K. Meijer, The action of flecainide acetate and its enantiomers on mammalian non-myelinated nerve fibres, Pharm Weekbl Sci 11 (1989) 92-94.

[0180] J. S. Lin, C. Anaclet, O. A. Sergeeva, H. L. Haas, The waking brain: an update, Cell Mol Life Sci 68 (2011) 2499-2512.

[0181] Lin J S, Dauvilliers Y, Arnulf I, Bastuji H, Anaclet C, Parmentier R, Kocher L, Yanagisawa M, Lehert P, Ligneau X, Perrin D, Robert P, Roux M, Lecomte J M, Schwartz J C (2008) An inverse agonist of the histamine H(3) receptor improves wakefulness in narcolepsy: studies in orexin-/- mice and patients. Neurobiology of disease 30: 74-83

[0182] Lin J S, Sergeeva O A, Haas H L (2011) Histamine H3 receptors and sleep-wake regulation. The Journal of pharmacology and experimental therapeutics 336: 17-23

[0183] Lin L, Faraco J, Li R, Kadotani H, Rogers W, Lin X, Qiu X, de Jong P J, Nishino S, Mignot E (1999) The sleep disorder canine narcolepsy is caused by a mutation in the hypocretin (orexin) receptor 2 gene. Cell 98: 365-376

[0184] R. Mehvar, D. R. Brocks, M. Vakily, Impact of stereoselectivity on the pharmacokinetics and pharmacodynamics of antiarrhythmic drugs, Clin Pharmacokinet 41 (2002) 533-558.

[0185] E. Mignot, S. Nishino, Emerging therapies in narcolepsy-cataplexy, Sleep 28 (2005) 754-763.

[0186] Minzenberg M J, Carter C S (2008) Modafinil: a review of neurochemical actions and effects on cognition. Neuropsychopharmacology 33: 1477-1502

[0187] Nakase T, Naus C C (2004) Gap junctions and neurological disorders of the central nervous system. Biochim Biophys Acta 1662: 149-158

[0188] R. Parmentier, H. Ohtsu, Z. Djebbara-Hannas, J. L. Valatx, T. Watanabe, J. S. Lin, Anatomical, physiological, and pharmacological characteristics of histidine decarboxylase knock-out mice: evidence for the role of brain histamine in behavioral and sleep-wake control, J Neurosci 22 (2002) 7695-7711.

[0189] Patel S J, Milwid J M, King K R, Bohr S, Iracheta-Velle A, Li M, Vitalo A, Parekkadan B, Jindal R, Yarmush M L (2012) Gap junction inhibition prevents drug-induced liver toxicity and fulminant hepatic failure. Nature biotechnology 30: 179-183

[0190] Sakurai T, Amemiya A, Ishii M, Matsuzaki I, Chemelli R M, Tanaka H, Williams S C, Richarson J A, Kozlowski G P, Wilson S, Arch J R, Buckingham R E, Haynes A C, Can S A, Annan R S, McNulty D E, Liu W S, Terrett J A, Elshourbagy N A, Bergsma D J, Yanagisawa M (1998) Orexins and orexin receptors: a family of hypothalamic neuropeptides and G protein-coupled receptors that regulate feeding behavior. Cell 92: 1 page following 696

[0191] Simon P, Panissaud C, Costentin J (1994) The stimulant effect of modafinil on wakefulness is not associated with an increase in anxiety in mice. A comparison with dexamphetamine. Psychopharmacology (Berl) 114: 597-600

[0192] J. K. Smallwood, D. W. Robertson, M. I. Steinberg, Electrophysiological effects of flecainide enantiomers in canine Purkinje fibres, Naunyn Schmiedebergs Arch Pharmacol 339 (1989) 625-629.

[0193] Takeuchi H, Mizoguchi H, Doi Y, Jin S, Noda M, Liang J, Li H, Zhou Y, Mori R, Yasuoka S, Li E, Parajuli B, Kawanokuchi J, Sonobe Y, Sato J, Yamanaka K, Sobue G, Mizuno T, Suzumura A (2011) Blockade of gap junction hemichannel suppresses disease progression in mouse models of amyotrophic lateral sclerosis and Alzheimer's disease. PLoS One 6: e21108

[0194] Tong X, Dong S, Yu M, Wang Q, Tao L (2013) Role of heteromeric gap junctions in the cytotoxicity of cisplatin. Toxicology 310C: 53-60

[0195] Traynelis S F, Wollmuth L P, McBain C J, Menniti F S, Vance K M, Ogden K K, Hansen K B, Yuan H, Myers S J, Dingledine R (2010) Glutamate receptor ion channels: structure, regulation, and function. Pharmacological reviews 62: 405-496

[0196] J. Turgeon, H. K. Kroemer, C. Prakash, I. A. Blair, D. M. Roden, Stereoselective determination of flecainide in human plasma by high-performance liquid chromatography with fluorescence detection, J Pharm Sci 79 (1990) 91-95.

Sequence CWU 1

1

211205PRThomo sapiensMISC_FEATURECx23 (A6NN92) 1Met Ser Leu Asn Tyr Ile Lys Asn Phe Tyr Glu Gly Cys Val Lys Pro 1 5 10 15 Pro Thr Val Ile Gly Gln Phe His Thr Leu Phe Phe Gly Ser Ile Arg 20 25 30 Ile Phe Phe Leu Gly Val Leu Gly Phe Ala Val Tyr Gly Asn Glu Ala 35 40 45 Leu His Phe Ile Cys Asp Pro Asp Lys Arg Glu Val Asn Leu Phe Cys 50 55 60 Tyr Asn Gln Phe Arg Pro Ile Thr Pro Gln Val Ser Phe Ser Ala Leu 65 70 75 80 Gln Leu Val Ile Val Leu Val Pro Gly Ala Leu Phe His Leu Tyr Ala 85 90 95 Ala Cys Lys Ser Ile Asn Gln Glu Cys Ile Leu Gln Lys Pro Ile Tyr 100 105 110 Thr Ile Ile Tyr Ile Leu Ser Val Leu Leu Arg Ile Ser Leu Ala Ala 115 120 125 Ile Ala Phe Trp Leu Gln Ile Tyr Leu Phe Gly Phe Gln Val Lys Ser 130 135 140 Leu Tyr Leu Cys Asp Ala Arg Ser Leu Gly Glu Asn Met Ile Ile Arg 145 150 155 160 Cys Met Val Pro Glu His Phe Glu Lys Thr Ile Phe Leu Ile Ala Ile 165 170 175 Asn Thr Phe Thr Thr Ile Thr Ile Leu Leu Phe Val Ala Glu Ile Phe 180 185 190 Glu Ile Ile Phe Arg Arg Leu Tyr Phe Pro Phe Arg Gln 195 200 205 2223PRThomo sapiensMISC_FEATURECx25 (NP_940970.1) 2Met Ser Trp Met Phe Leu Arg Asp Leu Leu Ser Gly Val Asn Lys Tyr 1 5 10 15 Ser Thr Gly Thr Gly Trp Ile Trp Leu Ala Val Val Phe Val Phe Arg 20 25 30 Leu Leu Val Tyr Met Val Ala Ala Glu His Val Trp Lys Asp Glu Gln 35 40 45 Lys Glu Phe Glu Cys Asn Ser Arg Gln Pro Gly Cys Lys Asn Val Cys 50 55 60 Phe Asp Asp Phe Phe Pro Ile Ser Gln Val Arg Leu Trp Ala Leu Gln 65 70 75 80 Leu Ile Met Val Ser Thr Pro Ser Leu Leu Val Val Leu His Val Ala 85 90 95 Tyr His Glu Gly Arg Glu Lys Arg His Arg Lys Lys Leu Tyr Val Ser 100 105 110 Pro Gly Thr Met Asp Gly Gly Leu Trp Tyr Ala Tyr Leu Ile Ser Leu 115 120 125 Ile Val Lys Thr Gly Phe Glu Ile Gly Phe Leu Val Leu Phe Tyr Lys 130 135 140 Leu Tyr Asp Gly Phe Ser Val Pro Tyr Leu Ile Lys Cys Asp Leu Lys 145 150 155 160 Pro Cys Pro Asn Thr Val Asp Cys Phe Ile Ser Lys Pro Thr Glu Lys 165 170 175 Thr Ile Phe Ile Leu Phe Leu Val Ile Thr Ser Cys Leu Cys Ile Val 180 185 190 Leu Asn Phe Ile Glu Leu Ser Phe Leu Val Leu Lys Cys Phe Ile Lys 195 200 205 Cys Cys Leu Gln Lys Tyr Leu Lys Lys Pro Gln Val Leu Ser Val 210 215 220 3226PRThomo sapiensMISC_FEATURECx26 (P29033.3) 3Met Asp Trp Gly Thr Leu Gln Thr Ile Leu Gly Gly Val Asn Lys His 1 5 10 15 Ser Thr Ser Ile Gly Lys Ile Trp Leu Thr Val Leu Phe Ile Phe Arg 20 25 30 Ile Met Ile Leu Val Val Ala Ala Lys Glu Val Trp Gly Asp Glu Gln 35 40 45 Ala Asp Phe Val Cys Asn Thr Leu Gln Pro Gly Cys Lys Asn Val Cys 50 55 60 Tyr Asp His Tyr Phe Pro Ile Ser His Ile Arg Leu Trp Ala Leu Gln 65 70 75 80 Leu Ile Phe Val Ser Thr Pro Ala Leu Leu Val Ala Met His Val Ala 85 90 95 Tyr Arg Arg His Glu Lys Lys Arg Lys Phe Ile Lys Gly Glu Ile Lys 100 105 110 Ser Glu Phe Lys Asp Ile Glu Glu Ile Lys Thr Gln Lys Val Arg Ile 115 120 125 Glu Gly Ser Leu Trp Trp Thr Tyr Thr Ser Ser Ile Phe Phe Arg Val 130 135 140 Ile Phe Glu Ala Ala Phe Met Tyr Val Phe Tyr Val Met Tyr Asp Gly 145 150 155 160 Phe Ser Met Gln Arg Leu Val Lys Cys Asn Ala Trp Pro Cys Pro Asn 165 170 175 Thr Val Asp Cys Phe Val Ser Arg Pro Thr Glu Lys Thr Val Phe Thr 180 185 190 Val Phe Met Ile Ala Val Ser Gly Ile Cys Ile Leu Leu Asn Val Thr 195 200 205 Glu Leu Cys Tyr Leu Leu Ile Arg Tyr Cys Ser Gly Lys Ser Lys Lys 210 215 220 Pro Val 225 4261PRThomo sapiensMISC_FEATURECx30 (NP_001103689.1) 4Met Asp Trp Gly Thr Leu His Thr Phe Ile Gly Gly Val Asn Lys His 1 5 10 15 Ser Thr Ser Ile Gly Lys Val Trp Ile Thr Val Ile Phe Ile Phe Arg 20 25 30 Val Met Ile Leu Val Val Ala Ala Gln Glu Val Trp Gly Asp Glu Gln 35 40 45 Glu Asp Phe Val Cys Asn Thr Leu Gln Pro Gly Cys Lys Asn Val Cys 50 55 60 Tyr Asp His Phe Phe Pro Val Ser His Ile Arg Leu Trp Ala Leu Gln 65 70 75 80 Leu Ile Phe Val Ser Thr Pro Ala Leu Leu Val Ala Met His Val Ala 85 90 95 Tyr Tyr Arg His Glu Thr Thr Arg Lys Phe Arg Arg Gly Glu Lys Arg 100 105 110 Asn Asp Phe Lys Asp Ile Glu Asp Ile Lys Lys Gln Lys Val Arg Ile 115 120 125 Glu Gly Ser Leu Trp Trp Thr Tyr Thr Ser Ser Ile Phe Phe Arg Ile 130 135 140 Ile Phe Glu Ala Ala Phe Met Tyr Val Phe Tyr Phe Leu Tyr Asn Gly 145 150 155 160 Tyr His Leu Pro Trp Val Leu Lys Cys Gly Ile Asp Pro Cys Pro Asn 165 170 175 Leu Val Asp Cys Phe Ile Ser Arg Pro Thr Glu Lys Thr Val Phe Thr 180 185 190 Ile Phe Met Ile Ser Ala Ser Val Ile Cys Met Leu Leu Asn Val Ala 195 200 205 Glu Leu Cys Tyr Leu Leu Leu Lys Val Cys Phe Arg Arg Ser Lys Arg 210 215 220 Ala Gln Thr Gln Lys Asn His Pro Asn His Ala Leu Lys Glu Ser Lys 225 230 235 240 Gln Asn Glu Met Asn Glu Leu Ile Ser Asp Ser Gly Gln Asn Ala Ile 245 250 255 Thr Gly Phe Pro Ser 260 5279PRThomo sapiensMISC_FEATURECx30.2 (NP_853516.1) 5Met Cys Gly Arg Phe Leu Arg Arg Leu Leu Ala Glu Glu Ser Arg Arg 1 5 10 15 Ser Thr Pro Val Gly Arg Leu Leu Leu Pro Val Leu Leu Gly Phe Arg 20 25 30 Leu Val Leu Leu Ala Ala Ser Gly Pro Gly Val Tyr Gly Asp Glu Gln 35 40 45 Ser Glu Phe Val Cys His Thr Gln Gln Pro Gly Cys Lys Ala Ala Cys 50 55 60 Phe Asp Ala Phe His Pro Leu Ser Pro Leu Arg Phe Trp Val Phe Gln 65 70 75 80 Val Ile Leu Val Ala Val Pro Ser Ala Leu Tyr Met Gly Phe Thr Leu 85 90 95 Tyr His Val Ile Trp His Trp Glu Leu Ser Gly Lys Gly Lys Glu Glu 100 105 110 Glu Thr Leu Ile Gln Gly Arg Glu Gly Asn Thr Asp Val Pro Gly Ala 115 120 125 Gly Ser Leu Arg Leu Leu Trp Ala Tyr Val Ala Gln Leu Gly Ala Arg 130 135 140 Leu Val Leu Glu Gly Ala Ala Leu Gly Leu Gln Tyr His Leu Tyr Gly 145 150 155 160 Phe Gln Met Pro Ser Ser Phe Ala Cys Arg Arg Glu Pro Cys Leu Gly 165 170 175 Ser Ile Thr Cys Asn Leu Ser Arg Pro Ser Glu Lys Thr Ile Phe Leu 180 185 190 Lys Thr Met Phe Gly Val Ser Gly Phe Cys Leu Leu Phe Thr Phe Leu 195 200 205 Glu Leu Val Leu Leu Gly Leu Gly Arg Trp Trp Arg Thr Trp Lys His 210 215 220 Lys Ser Ser Ser Ser Lys Tyr Phe Leu Thr Ser Glu Ser Thr Arg Arg 225 230 235 240 His Lys Lys Ala Thr Asp Ser Leu Pro Val Val Glu Thr Lys Glu Gln 245 250 255 Phe Gln Glu Ala Val Pro Gly Arg Ser Leu Ala Gln Glu Lys Gln Arg 260 265 270 Pro Val Gly Pro Arg Asp Ala 275 6266PRThomo sapiensMISC_FEATURECx30.3 (NP_694944.1) 6Met Asn Trp Ala Phe Leu Gln Gly Leu Leu Ser Gly Val Asn Lys Tyr 1 5 10 15 Ser Thr Val Leu Ser Arg Ile Trp Leu Ser Val Val Phe Ile Phe Arg 20 25 30 Val Leu Val Tyr Val Val Ala Ala Glu Glu Val Trp Asp Asp Glu Gln 35 40 45 Lys Asp Phe Val Cys Asn Thr Lys Gln Pro Gly Cys Pro Asn Val Cys 50 55 60 Tyr Asp Glu Phe Phe Pro Val Ser His Val Arg Leu Trp Ala Leu Gln 65 70 75 80 Leu Ile Leu Val Thr Cys Pro Ser Leu Leu Val Val Met His Val Ala 85 90 95 Tyr Arg Glu Glu Arg Glu Arg Lys His His Leu Lys His Gly Pro Asn 100 105 110 Ala Pro Ser Leu Tyr Asp Asn Leu Ser Lys Lys Arg Gly Gly Leu Trp 115 120 125 Trp Thr Tyr Leu Leu Ser Leu Ile Phe Lys Ala Ala Val Asp Ala Gly 130 135 140 Phe Leu Tyr Ile Phe His Arg Leu Tyr Lys Asp Tyr Asp Met Pro Arg 145 150 155 160 Val Val Ala Cys Ser Val Glu Pro Cys Pro His Thr Val Asp Cys Tyr 165 170 175 Ile Ser Arg Pro Thr Glu Lys Lys Val Phe Thr Tyr Phe Met Val Thr 180 185 190 Thr Ala Ala Ile Cys Ile Leu Leu Asn Leu Ser Glu Val Phe Tyr Leu 195 200 205 Val Gly Lys Arg Cys Met Glu Ile Phe Gly Pro Arg His Arg Arg Pro 210 215 220 Arg Cys Arg Glu Cys Leu Pro Asp Thr Cys Pro Pro Tyr Val Leu Ser 225 230 235 240 Gln Gly Gly His Pro Glu Asp Gly Asn Ser Val Leu Met Lys Ala Gly 245 250 255 Ser Ala Pro Val Asp Ala Gly Gly Tyr Pro 260 265 7270PRThomo sapiensMISC_FEATURECx31 (NP_001005752.1) 7Met Asp Trp Lys Thr Leu Gln Ala Leu Leu Ser Gly Val Asn Lys Tyr 1 5 10 15 Ser Thr Ala Phe Gly Arg Ile Trp Leu Ser Val Val Phe Val Phe Arg 20 25 30 Val Leu Val Tyr Val Val Ala Ala Glu Arg Val Trp Gly Asp Glu Gln 35 40 45 Lys Asp Phe Asp Cys Asn Thr Lys Gln Pro Gly Cys Thr Asn Val Cys 50 55 60 Tyr Asp Asn Tyr Phe Pro Ile Ser Asn Ile Arg Leu Trp Ala Leu Gln 65 70 75 80 Leu Ile Phe Val Thr Cys Pro Ser Leu Leu Val Ile Leu His Val Ala 85 90 95 Tyr Arg Glu Glu Arg Glu Arg Arg His Arg Gln Lys His Gly Asp Gln 100 105 110 Cys Ala Lys Leu Tyr Asp Asn Ala Gly Lys Lys His Gly Gly Leu Trp 115 120 125 Trp Thr Tyr Leu Phe Ser Leu Ile Phe Lys Leu Ile Ile Glu Phe Leu 130 135 140 Phe Leu Tyr Leu Leu His Thr Leu Trp His Gly Phe Asn Met Pro Arg 145 150 155 160 Leu Val Gln Cys Ala Asn Val Ala Pro Cys Pro Asn Ile Val Asp Cys 165 170 175 Tyr Ile Ala Arg Pro Thr Glu Lys Lys Ile Phe Thr Tyr Phe Met Val 180 185 190 Gly Ala Ser Ala Val Cys Ile Val Leu Thr Ile Cys Glu Leu Cys Tyr 195 200 205 Leu Ile Cys His Arg Val Leu Arg Gly Leu His Lys Asp Lys Pro Arg 210 215 220 Gly Gly Cys Ser Pro Ser Ser Ser Ala Ser Arg Ala Ser Thr Cys Arg 225 230 235 240 Cys His His Lys Leu Val Glu Ala Gly Glu Val Asp Pro Asp Pro Gly 245 250 255 Asn Asn Lys Leu Gln Ala Ser Ala Pro Asn Leu Thr Pro Ile 260 265 270 8273PRThomo sapiensMISC_FEATURECx31.1 (NP_005259.1) 8Met Asn Trp Ser Ile Phe Glu Gly Leu Leu Ser Gly Val Asn Lys Tyr 1 5 10 15 Ser Thr Ala Phe Gly Arg Ile Trp Leu Ser Leu Val Phe Ile Phe Arg 20 25 30 Val Leu Val Tyr Leu Val Thr Ala Glu Arg Val Trp Ser Asp Asp His 35 40 45 Lys Asp Phe Asp Cys Asn Thr Arg Gln Pro Gly Cys Ser Asn Val Cys 50 55 60 Phe Asp Glu Phe Phe Pro Val Ser His Val Arg Leu Trp Ala Leu Gln 65 70 75 80 Leu Ile Leu Val Thr Cys Pro Ser Leu Leu Val Val Met His Val Ala 85 90 95 Tyr Arg Glu Val Gln Glu Lys Arg His Arg Glu Ala His Gly Glu Asn 100 105 110 Ser Gly Arg Leu Tyr Leu Asn Pro Gly Lys Lys Arg Gly Gly Leu Trp 115 120 125 Trp Thr Tyr Val Cys Ser Leu Val Phe Lys Ala Ser Val Asp Ile Ala 130 135 140 Phe Leu Tyr Val Phe His Ser Phe Tyr Pro Lys Tyr Ile Leu Pro Pro 145 150 155 160 Val Val Lys Cys His Ala Asp Pro Cys Pro Asn Ile Val Asp Cys Phe 165 170 175 Ile Ser Lys Pro Ser Glu Lys Asn Ile Phe Thr Leu Phe Met Val Ala 180 185 190 Thr Ala Ala Ile Cys Ile Leu Leu Asn Leu Val Glu Leu Ile Tyr Leu 195 200 205 Val Ser Lys Arg Cys His Glu Cys Leu Ala Ala Arg Lys Ala Gln Ala 210 215 220 Met Cys Thr Gly His His Pro His Gly Thr Thr Ser Ser Cys Lys Gln 225 230 235 240 Asp Asp Leu Leu Ser Gly Asp Leu Ile Phe Leu Gly Ser Asp Ser His 245 250 255 Pro Pro Leu Leu Pro Asp Arg Pro Arg Asp His Val Lys Lys Thr Ile 260 265 270 Leu 9294PRThomo sapiensmisc_featureCx31.9 (NP_689343.3) 9Met Gly Glu Trp Ala Phe Leu Gly Ser Leu Leu Asp Ala Val Gln Leu 1 5 10 15 Gln Ser Pro Leu Val Gly Arg Leu Trp Leu Val Val Met Leu Ile Phe 20 25 30 Arg Ile Leu Val Leu Ala Thr Val Gly Gly Ala Val Phe Glu Asp Glu 35 40 45 Gln Glu Glu Phe Val Cys Asn Thr Leu Gln Pro Gly Cys Arg Gln Thr 50 55 60 Cys Tyr Asp Arg Ala Phe Pro Val Ser His Tyr Arg Phe Trp Leu Phe 65 70 75 80 His Ile Leu Leu Leu Ser Ala Pro Pro Val Leu Phe Val Val Tyr Ser 85 90 95 Met His Arg Ala Gly Lys Glu Ala Gly Gly Ala Glu Ala Ala Ala Gln 100 105 110 Cys Ala Pro Gly Leu Pro Glu Ala Gln Cys Ala Pro Cys Ala Leu Arg 115 120 125 Ala Arg Arg Ala Arg Arg Cys Tyr Leu Leu Ser Val Ala Leu Arg Leu 130 135 140 Leu Ala Glu Leu Thr Phe Leu Gly Gly Gln Ala Leu Leu Tyr Gly Phe 145 150 155 160 Arg Val Ala Pro His Phe Ala Cys Ala Gly Pro Pro Cys Pro His Thr 165 170 175 Val Asp Cys Phe Val Ser Arg Pro Thr Glu Lys Thr Val Phe Val Leu 180 185 190 Phe Tyr Phe Ala Val Gly Leu Leu Ser Ala Leu Leu Ser Val Ala Glu 195 200 205 Leu Gly His Leu Leu Trp Lys Gly Arg Pro Arg Ala Gly Glu Arg Asp 210 215 220 Asn Arg Cys Asn Arg Ala His Glu Glu Ala Gln Lys Leu Leu Pro Pro 225 230 235

240 Pro Pro Pro Pro Pro Pro Pro Pro Ala Leu Pro Ser Arg Arg Pro Gly 245 250 255 Pro Glu Pro Cys Ala Pro Pro Ala Tyr Ala His Pro Ala Pro Ala Ser 260 265 270 Leu Arg Glu Cys Gly Ser Gly Arg Gly Lys Ala Ser Pro Ala Thr Gly 275 280 285 Arg Arg Asp Leu Ala Ile 290 10283PRThomo sapiensMISC_FEATURECx32 (NP_000157.1) 10Met Asn Trp Thr Gly Leu Tyr Thr Leu Leu Ser Gly Val Asn Arg His 1 5 10 15 Ser Thr Ala Ile Gly Arg Val Trp Leu Ser Val Ile Phe Ile Phe Arg 20 25 30 Ile Met Val Leu Val Val Ala Ala Glu Ser Val Trp Gly Asp Glu Lys 35 40 45 Ser Ser Phe Ile Cys Asn Thr Leu Gln Pro Gly Cys Asn Ser Val Cys 50 55 60 Tyr Asp Gln Phe Phe Pro Ile Ser His Val Arg Leu Trp Ser Leu Gln 65 70 75 80 Leu Ile Leu Val Ser Thr Pro Ala Leu Leu Val Ala Met His Val Ala 85 90 95 His Gln Gln His Ile Glu Lys Lys Met Leu Arg Leu Glu Gly His Gly 100 105 110 Asp Pro Leu His Leu Glu Glu Val Lys Arg His Lys Val His Ile Ser 115 120 125 Gly Thr Leu Trp Trp Thr Tyr Val Ile Ser Val Val Phe Arg Leu Leu 130 135 140 Phe Glu Ala Val Phe Met Tyr Val Phe Tyr Leu Leu Tyr Pro Gly Tyr 145 150 155 160 Ala Met Val Arg Leu Val Lys Cys Asp Val Tyr Pro Cys Pro Asn Thr 165 170 175 Val Asp Cys Phe Val Ser Arg Pro Thr Glu Lys Thr Val Phe Thr Val 180 185 190 Phe Met Leu Ala Ala Ser Gly Ile Cys Ile Ile Leu Asn Val Ala Glu 195 200 205 Val Val Tyr Leu Ile Ile Arg Ala Cys Ala Arg Arg Ala Gln Arg Arg 210 215 220 Ser Asn Pro Pro Ser Arg Lys Gly Ser Gly Phe Gly His Arg Leu Ser 225 230 235 240 Pro Glu Tyr Lys Gln Asn Glu Ile Asn Lys Leu Leu Ser Glu Gln Asp 245 250 255 Gly Ser Leu Lys Asp Ile Leu Arg Arg Ser Pro Gly Thr Gly Ala Gly 260 265 270 Leu Ala Glu Lys Ser Asp Arg Cys Ser Ala Cys 275 280 11321PRThomo sapiensMISC_FEATURECx36 (NP_065711.1) 11Met Gly Glu Trp Thr Ile Leu Glu Arg Leu Leu Glu Ala Ala Val Gln 1 5 10 15 Gln His Ser Thr Met Ile Gly Arg Ile Leu Leu Thr Val Val Val Ile 20 25 30 Phe Arg Ile Leu Ile Val Ala Ile Val Gly Glu Thr Val Tyr Asp Asp 35 40 45 Glu Gln Thr Met Phe Val Cys Asn Thr Leu Gln Pro Gly Cys Asn Gln 50 55 60 Ala Cys Tyr Asp Arg Ala Phe Pro Ile Ser His Ile Arg Tyr Trp Val 65 70 75 80 Phe Gln Ile Ile Met Val Cys Thr Pro Ser Leu Cys Phe Ile Thr Tyr 85 90 95 Ser Val His Gln Ser Ala Lys Gln Arg Glu Arg Arg Tyr Ser Thr Val 100 105 110 Phe Leu Ala Leu Asp Arg Asp Pro Pro Glu Ser Ile Gly Gly Pro Gly 115 120 125 Gly Thr Gly Gly Gly Gly Ser Gly Gly Gly Lys Arg Glu Asp Lys Lys 130 135 140 Leu Gln Asn Ala Ile Val Asn Gly Val Leu Gln Asn Thr Glu Asn Thr 145 150 155 160 Ser Lys Glu Thr Glu Pro Asp Cys Leu Glu Val Lys Glu Leu Thr Pro 165 170 175 His Pro Ser Gly Leu Arg Thr Ala Ser Lys Ser Lys Leu Arg Arg Gln 180 185 190 Glu Gly Ile Ser Arg Phe Tyr Ile Ile Gln Val Val Phe Arg Asn Ala 195 200 205 Leu Glu Ile Gly Phe Leu Val Gly Gln Tyr Phe Leu Tyr Gly Phe Ser 210 215 220 Val Pro Gly Leu Tyr Glu Cys Asn Arg Tyr Pro Cys Ile Lys Glu Val 225 230 235 240 Glu Cys Tyr Val Ser Arg Pro Thr Glu Lys Thr Val Phe Leu Val Phe 245 250 255 Met Phe Ala Val Ser Gly Ile Cys Val Val Leu Asn Leu Ala Glu Leu 260 265 270 Asn His Leu Gly Trp Arg Lys Ile Lys Leu Ala Val Arg Gly Ala Gln 275 280 285 Ala Lys Arg Lys Ser Ile Tyr Glu Ile Arg Asn Lys Asp Leu Pro Arg 290 295 300 Val Ser Val Pro Asn Phe Gly Arg Thr Gln Ser Ser Asp Ser Ala Tyr 305 310 315 320 Val 12333PRThomo sapiensMISC_FEATURECx37 (NP_002051.2) 12Met Gly Asp Trp Gly Phe Leu Glu Lys Leu Leu Asp Gln Val Gln Glu 1 5 10 15 His Ser Thr Val Val Gly Lys Ile Trp Leu Thr Val Leu Phe Ile Phe 20 25 30 Arg Ile Leu Ile Leu Gly Leu Ala Gly Glu Ser Val Trp Gly Asp Glu 35 40 45 Gln Ser Asp Phe Glu Cys Asn Thr Ala Gln Pro Gly Cys Thr Asn Val 50 55 60 Cys Tyr Asp Gln Ala Phe Pro Ile Ser His Ile Arg Tyr Trp Val Leu 65 70 75 80 Gln Phe Leu Phe Val Ser Thr Pro Thr Leu Val Tyr Leu Gly His Val 85 90 95 Ile Tyr Leu Ser Arg Arg Glu Glu Arg Leu Arg Gln Lys Glu Gly Glu 100 105 110 Leu Arg Ala Leu Pro Ala Lys Asp Pro Gln Val Glu Arg Ala Leu Ala 115 120 125 Ala Val Glu Arg Gln Met Ala Lys Ile Ser Val Ala Glu Asp Gly Arg 130 135 140 Leu Arg Ile Arg Gly Ala Leu Met Gly Thr Tyr Val Ala Ser Val Leu 145 150 155 160 Cys Lys Ser Val Leu Glu Ala Gly Phe Leu Tyr Gly Gln Trp Arg Leu 165 170 175 Tyr Gly Trp Thr Met Glu Pro Val Phe Val Cys Gln Arg Ala Pro Cys 180 185 190 Pro Tyr Leu Val Asp Cys Phe Val Ser Arg Pro Thr Glu Lys Thr Ile 195 200 205 Phe Ile Ile Phe Met Leu Val Val Gly Leu Ile Ser Leu Val Leu Asn 210 215 220 Leu Leu Glu Leu Val His Leu Leu Cys Arg Cys Leu Ser Arg Gly Met 225 230 235 240 Arg Ala Arg Gln Gly Gln Asp Ala Pro Pro Thr Gln Gly Thr Ser Ser 245 250 255 Asp Pro Tyr Thr Asp Gln Val Phe Phe Tyr Leu Pro Val Gly Gln Gly 260 265 270 Pro Ser Ser Pro Pro Cys Pro Thr Tyr Asn Gly Leu Ser Ser Ser Glu 275 280 285 Gln Asn Trp Ala Asn Leu Thr Thr Glu Glu Arg Leu Ala Ser Ser Arg 290 295 300 Pro Pro Leu Phe Leu Asp Pro Pro Pro Gln Asn Gly Gln Lys Pro Pro 305 310 315 320 Ser Arg Pro Ser Ser Ser Ala Ser Lys Lys Gln Tyr Val 325 330 13358PRThomo sapiensMISC_FEATURECx40 (NP_005257) 13Met Gly Asp Trp Ser Phe Leu Gly Asn Phe Leu Glu Glu Val His Lys 1 5 10 15 His Ser Thr Val Val Gly Lys Val Trp Leu Thr Val Leu Phe Ile Phe 20 25 30 Arg Met Leu Val Leu Gly Thr Ala Ala Glu Ser Ser Trp Gly Asp Glu 35 40 45 Gln Ala Asp Phe Arg Cys Asp Thr Ile Gln Pro Gly Cys Gln Asn Val 50 55 60 Cys Tyr Asp Gln Ala Phe Pro Ile Ser His Ile Arg Tyr Trp Val Leu 65 70 75 80 Gln Ile Ile Phe Val Ser Thr Pro Ser Leu Val Tyr Met Gly His Ala 85 90 95 Met His Thr Val Arg Met Gln Glu Lys Arg Lys Leu Arg Glu Ala Glu 100 105 110 Arg Ala Lys Glu Val Arg Gly Ser Gly Ser Tyr Glu Tyr Pro Val Ala 115 120 125 Glu Lys Ala Glu Leu Ser Cys Trp Glu Glu Gly Asn Gly Arg Ile Ala 130 135 140 Leu Gln Gly Thr Leu Leu Asn Thr Tyr Val Cys Ser Ile Leu Ile Arg 145 150 155 160 Thr Thr Met Glu Val Gly Phe Ile Val Gly Gln Tyr Phe Ile Tyr Gly 165 170 175 Ile Phe Leu Thr Thr Leu His Val Cys Arg Arg Ser Pro Cys Pro His 180 185 190 Pro Val Asn Cys Tyr Val Ser Arg Pro Thr Glu Lys Asn Val Phe Ile 195 200 205 Val Phe Met Leu Ala Val Ala Ala Leu Ser Leu Leu Leu Ser Leu Ala 210 215 220 Glu Leu Tyr His Leu Gly Trp Lys Lys Ile Arg Gln Arg Phe Val Lys 225 230 235 240 Pro Arg Gln His Met Ala Lys Cys Gln Leu Ser Gly Pro Ser Val Gly 245 250 255 Ile Val Gln Ser Cys Thr Pro Pro Pro Asp Phe Asn Gln Cys Leu Glu 260 265 270 Asn Gly Pro Gly Gly Lys Phe Phe Asn Pro Phe Ser Asn Asn Met Ala 275 280 285 Ser Gln Gln Asn Thr Asp Asn Leu Val Thr Glu Gln Val Arg Gly Gln 290 295 300 Glu Gln Thr Pro Gly Glu Gly Phe Ile Gln Val Arg Tyr Gly Gln Lys 305 310 315 320 Pro Glu Val Pro Asn Gly Val Ser Pro Gly His Arg Leu Pro His Gly 325 330 335 Tyr His Ser Asp Lys Arg Arg Leu Ser Lys Ala Ser Ser Lys Ala Arg 340 345 350 Ser Asp Asp Leu Ser Val 355 14370PRThomo sapiensmisc_featureCx40.1 (NP_699199.2) 14Met Glu Gly Val Asp Leu Leu Gly Phe Leu Ile Ile Thr Leu Asn Cys 1 5 10 15 Asn Val Thr Met Val Gly Lys Leu Trp Phe Val Leu Thr Met Leu Leu 20 25 30 Arg Met Leu Val Ile Val Leu Ala Gly Arg Pro Val Tyr Gln Asp Glu 35 40 45 Gln Glu Arg Phe Val Cys Asn Thr Leu Gln Pro Gly Cys Ala Asn Val 50 55 60 Cys Tyr Asp Val Phe Ser Pro Val Ser His Leu Arg Phe Trp Leu Ile 65 70 75 80 Gln Gly Val Cys Val Leu Leu Pro Ser Ala Val Phe Ser Val Tyr Val 85 90 95 Leu His Arg Gly Ala Thr Leu Ala Ala Leu Gly Pro Arg Arg Cys Pro 100 105 110 Asp Pro Arg Glu Pro Ala Ser Gly Gln Arg Arg Cys Pro Arg Pro Phe 115 120 125 Gly Glu Arg Gly Gly Leu Gln Val Pro Asp Phe Ser Ala Gly Tyr Ile 130 135 140 Ile His Leu Leu Leu Arg Thr Leu Leu Glu Ala Ala Phe Gly Ala Leu 145 150 155 160 His Tyr Phe Leu Phe Gly Phe Leu Ala Pro Lys Lys Phe Pro Cys Thr 165 170 175 Arg Pro Pro Cys Thr Gly Val Val Asp Cys Tyr Val Ser Arg Pro Thr 180 185 190 Glu Lys Ser Leu Leu Met Leu Phe Leu Trp Ala Val Ser Ala Leu Ser 195 200 205 Phe Leu Leu Gly Leu Ala Asp Leu Val Cys Ser Leu Arg Arg Arg Met 210 215 220 Arg Arg Arg Pro Gly Pro Pro Thr Ser Pro Ser Ile Arg Lys Gln Ser 225 230 235 240 Gly Ala Ser Gly His Ala Glu Gly Arg Arg Thr Asp Glu Glu Gly Gly 245 250 255 Arg Glu Glu Glu Gly Ala Pro Ala Pro Pro Gly Ala Arg Ala Gly Gly 260 265 270 Glu Gly Ala Gly Ser Pro Arg Arg Thr Ser Arg Val Ser Gly His Thr 275 280 285 Lys Ile Pro Asp Glu Asp Glu Ser Glu Val Thr Ser Ser Ala Ser Glu 290 295 300 Lys Leu Gly Arg Gln Pro Arg Gly Arg Pro His Arg Glu Ala Ala Gln 305 310 315 320 Asp Pro Arg Gly Ser Gly Ser Glu Glu Gln Pro Ser Ala Ala Pro Ser 325 330 335 Arg Leu Ala Ala Pro Pro Ser Cys Ser Ser Leu Gln Pro Pro Asp Pro 340 345 350 Pro Ala Ser Ser Ser Gly Ala Pro His Leu Arg Ala Arg Lys Ser Glu 355 360 365 Trp Val 370 15382PRThomo sapiensMISC_FEATURECx43 (NP_000156.1) 15Met Gly Asp Trp Ser Ala Leu Gly Lys Leu Leu Asp Lys Val Gln Ala 1 5 10 15 Tyr Ser Thr Ala Gly Gly Lys Val Trp Leu Ser Val Leu Phe Ile Phe 20 25 30 Arg Ile Leu Leu Leu Gly Thr Ala Val Glu Ser Ala Trp Gly Asp Glu 35 40 45 Gln Ser Ala Phe Arg Cys Asn Thr Gln Gln Pro Gly Cys Glu Asn Val 50 55 60 Cys Tyr Asp Lys Ser Phe Pro Ile Ser His Val Arg Phe Trp Val Leu 65 70 75 80 Gln Ile Ile Phe Val Ser Val Pro Thr Leu Leu Tyr Leu Ala His Val 85 90 95 Phe Tyr Val Met Arg Lys Glu Glu Lys Leu Asn Lys Lys Glu Glu Glu 100 105 110 Leu Lys Val Ala Gln Thr Asp Gly Val Asn Val Asp Met His Leu Lys 115 120 125 Gln Ile Glu Ile Lys Lys Phe Lys Tyr Gly Ile Glu Glu His Gly Lys 130 135 140 Val Lys Met Arg Gly Gly Leu Leu Arg Thr Tyr Ile Ile Ser Ile Leu 145 150 155 160 Phe Lys Ser Ile Phe Glu Val Ala Phe Leu Leu Ile Gln Trp Tyr Ile 165 170 175 Tyr Gly Phe Ser Leu Ser Ala Val Tyr Thr Cys Lys Arg Asp Pro Cys 180 185 190 Pro His Gln Val Asp Cys Phe Leu Ser Arg Pro Thr Glu Lys Thr Ile 195 200 205 Phe Ile Ile Phe Met Leu Val Val Ser Leu Val Ser Leu Ala Leu Asn 210 215 220 Ile Ile Glu Leu Phe Tyr Val Phe Phe Lys Gly Val Lys Asp Arg Val 225 230 235 240 Lys Gly Lys Ser Asp Pro Tyr His Ala Thr Ser Gly Ala Leu Ser Pro 245 250 255 Ala Lys Asp Cys Gly Ser Gln Lys Tyr Ala Tyr Phe Asn Gly Cys Ser 260 265 270 Ser Pro Thr Ala Pro Leu Ser Pro Met Ser Pro Pro Gly Tyr Lys Leu 275 280 285 Val Thr Gly Asp Arg Asn Asn Ser Ser Cys Arg Asn Tyr Asn Lys Gln 290 295 300 Ala Ser Glu Gln Asn Trp Ala Asn Tyr Ser Ala Glu Gln Asn Arg Met 305 310 315 320 Gly Gln Ala Gly Ser Thr Ile Ser Asn Ser His Ala Gln Pro Phe Asp 325 330 335 Phe Pro Asp Asp Asn Gln Asn Ser Lys Lys Leu Ala Ala Gly His Glu 340 345 350 Leu Gln Pro Leu Ala Ile Val Asp Gln Arg Pro Ser Ser Arg Ala Ser 355 360 365 Ser Arg Ala Ser Ser Arg Pro Arg Pro Asp Asp Leu Glu Ile 370 375 380 16396PRThomo sapiensMISC_FEATURECx45 (NP_001073852.1) 16Met Ser Trp Ser Phe Leu Thr Arg Leu Leu Glu Glu Ile His Asn His 1 5 10 15 Ser Thr Phe Val Gly Lys Ile Trp Leu Thr Val Leu Ile Val Phe Arg 20 25 30 Ile Val Leu Thr Ala Val Gly Gly Glu Ser Ile Tyr Tyr Asp Glu Gln 35 40 45 Ser Lys Phe Val Cys Asn Thr Glu Gln Pro Gly Cys Glu Asn Val Cys 50 55 60 Tyr Asp Ala Phe Ala Pro Leu Ser His Val Arg Phe Trp Val Phe Gln 65 70 75 80 Ile Ile Leu Val Ala Thr Pro Ser Val Met Tyr Leu Gly Tyr Ala Ile 85 90 95 His Lys Ile Ala Lys Met Glu His Gly Glu Ala Asp Lys Lys Ala Ala 100 105 110 Arg Ser Lys Pro Tyr Ala Met Arg Trp Lys Gln His Arg Ala Leu Glu 115 120 125 Glu Thr Glu Glu Asp Asn Glu Glu Asp Pro Met Met Tyr Pro Glu Met 130 135 140 Glu Leu Glu Ser Asp Lys Glu Asn Lys Glu Gln Ser Gln Pro Lys Pro 145 150

155 160 Lys His Asp Gly Arg Arg Arg Ile Arg Glu Asp Gly Leu Met Lys Ile 165 170 175 Tyr Val Leu Gln Leu Leu Ala Arg Thr Val Phe Glu Val Gly Phe Leu 180 185 190 Ile Gly Gln Tyr Phe Leu Tyr Gly Phe Gln Val His Pro Phe Tyr Val 195 200 205 Cys Ser Arg Leu Pro Cys Pro His Lys Ile Asp Cys Phe Ile Ser Arg 210 215 220 Pro Thr Glu Lys Thr Ile Phe Leu Leu Ile Met Tyr Gly Val Thr Gly 225 230 235 240 Leu Cys Leu Leu Leu Asn Ile Trp Glu Met Leu His Leu Gly Phe Gly 245 250 255 Thr Ile Arg Asp Ser Leu Asn Ser Lys Arg Arg Glu Leu Glu Asp Pro 260 265 270 Gly Ala Tyr Asn Tyr Pro Phe Thr Trp Asn Thr Pro Ser Ala Pro Pro 275 280 285 Gly Tyr Asn Ile Ala Val Lys Pro Asp Gln Ile Gln Tyr Thr Glu Leu 290 295 300 Ser Asn Ala Lys Ile Ala Tyr Lys Gln Asn Lys Ala Asn Thr Ala Gln 305 310 315 320 Glu Gln Gln Tyr Gly Ser His Glu Glu Asn Leu Pro Ala Asp Leu Glu 325 330 335 Ala Leu Gln Arg Glu Ile Arg Met Ala Gln Glu Arg Leu Asp Leu Ala 340 345 350 Val Gln Ala Tyr Ser His Gln Asn Asn Pro His Gly Pro Arg Glu Lys 355 360 365 Lys Ala Lys Val Gly Ser Lys Ala Gly Ser Asn Lys Ser Thr Ala Ser 370 375 380 Ser Lys Ser Gly Asp Gly Lys Thr Ser Val Trp Ile 385 390 395 17435PRThomo sapiensMISC_FEATURECx46 (NP_068773.2) 17Met Gly Asp Trp Ser Phe Leu Gly Arg Leu Leu Glu Asn Ala Gln Glu 1 5 10 15 His Ser Thr Val Ile Gly Lys Val Trp Leu Thr Val Leu Phe Ile Phe 20 25 30 Arg Ile Leu Val Leu Gly Ala Ala Ala Glu Asp Val Trp Gly Asp Glu 35 40 45 Gln Ser Asp Phe Thr Cys Asn Thr Gln Gln Pro Gly Cys Glu Asn Val 50 55 60 Cys Tyr Asp Arg Ala Phe Pro Ile Ser His Ile Arg Phe Trp Ala Leu 65 70 75 80 Gln Ile Ile Phe Val Ser Thr Pro Thr Leu Ile Tyr Leu Gly His Val 85 90 95 Leu His Ile Val Arg Met Glu Glu Lys Lys Lys Glu Arg Glu Glu Glu 100 105 110 Glu Gln Leu Lys Arg Glu Ser Pro Ser Pro Lys Glu Pro Pro Gln Asp 115 120 125 Asn Pro Ser Ser Arg Asp Asp Arg Gly Arg Val Arg Met Ala Gly Ala 130 135 140 Leu Leu Arg Thr Tyr Val Phe Asn Ile Ile Phe Lys Thr Leu Phe Glu 145 150 155 160 Val Gly Phe Ile Ala Gly Gln Tyr Phe Leu Tyr Gly Phe Glu Leu Lys 165 170 175 Pro Leu Tyr Arg Cys Asp Arg Trp Pro Cys Pro Asn Thr Val Asp Cys 180 185 190 Phe Ile Ser Arg Pro Thr Glu Lys Thr Ile Phe Ile Ile Phe Met Leu 195 200 205 Ala Val Ala Cys Ala Ser Leu Leu Leu Asn Met Leu Glu Ile Tyr His 210 215 220 Leu Gly Trp Lys Lys Leu Lys Gln Gly Val Thr Ser Arg Leu Gly Pro 225 230 235 240 Asp Ala Ser Glu Ala Pro Leu Gly Thr Ala Asp Pro Pro Pro Leu Pro 245 250 255 Pro Ser Ser Arg Pro Pro Ala Val Ala Ile Gly Phe Pro Pro Tyr Tyr 260 265 270 Ala His Thr Ala Ala Pro Leu Gly Gln Ala Arg Ala Val Gly Tyr Pro 275 280 285 Gly Ala Pro Pro Pro Ala Ala Asp Phe Lys Leu Leu Ala Leu Thr Glu 290 295 300 Ala Arg Gly Lys Gly Gln Ser Ala Lys Leu Tyr Asn Gly His His His 305 310 315 320 Leu Leu Met Thr Glu Gln Asn Trp Ala Asn Gln Ala Ala Glu Arg Gln 325 330 335 Pro Pro Ala Leu Lys Ala Tyr Pro Ala Ala Ser Thr Pro Ala Ala Pro 340 345 350 Ser Pro Val Gly Ser Ser Ser Pro Pro Leu Ala His Glu Ala Glu Ala 355 360 365 Gly Ala Ala Pro Leu Leu Leu Asp Gly Ser Gly Ser Ser Leu Glu Gly 370 375 380 Ser Ala Leu Ala Gly Thr Pro Glu Glu Glu Glu Gln Ala Val Thr Thr 385 390 395 400 Ala Ala Gln Met His Gln Pro Pro Leu Pro Leu Gly Asp Pro Gly Arg 405 410 415 Ala Ser Lys Ala Ser Arg Ala Ser Ser Gly Arg Ala Arg Pro Glu Asp 420 425 430 Leu Ala Ile 435 18439PRThomo sapiensMISC_FEATURECx47 (NP_065168.2) 18Met Thr Asn Met Ser Trp Ser Phe Leu Thr Arg Leu Leu Glu Glu Ile 1 5 10 15 His Asn His Ser Thr Phe Val Gly Lys Val Trp Leu Thr Val Leu Val 20 25 30 Val Phe Arg Ile Val Leu Thr Ala Val Gly Gly Glu Ala Ile Tyr Ser 35 40 45 Asp Glu Gln Ala Lys Phe Thr Cys Asn Thr Arg Gln Pro Gly Cys Asp 50 55 60 Asn Val Cys Tyr Asp Ala Phe Ala Pro Leu Ser His Val Arg Phe Trp 65 70 75 80 Val Phe Gln Ile Val Val Ile Ser Thr Pro Ser Val Met Tyr Leu Gly 85 90 95 Tyr Ala Val His Arg Leu Ala Arg Ala Ser Glu Gln Glu Arg Arg Arg 100 105 110 Ala Leu Arg Arg Arg Pro Gly Pro Arg Arg Ala Pro Arg Ala His Leu 115 120 125 Pro Pro Pro His Ala Gly Trp Pro Glu Pro Ala Asp Leu Gly Glu Glu 130 135 140 Glu Pro Met Leu Gly Leu Gly Glu Glu Glu Glu Glu Glu Glu Thr Gly 145 150 155 160 Ala Ala Glu Gly Ala Gly Glu Glu Ala Glu Glu Ala Gly Ala Glu Glu 165 170 175 Ala Cys Thr Lys Ala Val Gly Ala Asp Gly Lys Ala Ala Gly Thr Pro 180 185 190 Gly Pro Thr Gly Gln His Asp Gly Arg Arg Arg Ile Gln Arg Glu Gly 195 200 205 Leu Met Arg Val Tyr Val Ala Gln Leu Val Ala Arg Ala Ala Phe Glu 210 215 220 Val Ala Phe Leu Val Gly Gln Tyr Leu Leu Tyr Gly Phe Glu Val Arg 225 230 235 240 Pro Phe Phe Pro Cys Ser Arg Gln Pro Cys Pro His Val Val Asp Cys 245 250 255 Phe Val Ser Arg Pro Thr Glu Lys Thr Val Phe Leu Leu Val Met Tyr 260 265 270 Val Val Ser Cys Leu Cys Leu Leu Leu Asn Leu Cys Glu Met Ala His 275 280 285 Leu Gly Leu Gly Ser Ala Gln Asp Ala Val Arg Gly Arg Arg Gly Pro 290 295 300 Pro Ala Ser Ala Pro Ala Pro Ala Pro Arg Pro Pro Pro Cys Ala Phe 305 310 315 320 Pro Ala Ala Ala Ala Gly Leu Ala Cys Pro Pro Asp Tyr Ser Leu Val 325 330 335 Val Arg Ala Ala Glu Arg Ala Arg Ala His Asp Gln Asn Leu Ala Asn 340 345 350 Leu Ala Leu Gln Ala Leu Arg Asp Gly Ala Ala Ala Gly Asp Arg Asp 355 360 365 Arg Asp Ser Ser Pro Cys Val Gly Leu Pro Ala Ala Ser Arg Gly Pro 370 375 380 Pro Arg Ala Gly Ala Pro Ala Ser Arg Thr Gly Ser Ala Thr Ser Ala 385 390 395 400 Gly Thr Val Gly Glu Gln Gly Arg Pro Gly Thr His Glu Arg Pro Gly 405 410 415 Ala Lys Pro Arg Ala Gly Ser Glu Lys Gly Ser Ala Ser Ser Arg Asp 420 425 430 Gly Lys Thr Thr Val Trp Ile 435 19433PRThomo sapiensMISC_FEATURECx50 (NP_005258.2) 19Met Gly Asp Trp Ser Phe Leu Gly Asn Ile Leu Glu Glu Val Asn Glu 1 5 10 15 His Ser Thr Val Ile Gly Arg Val Trp Leu Thr Val Leu Phe Ile Phe 20 25 30 Arg Ile Leu Ile Leu Gly Thr Ala Ala Glu Phe Val Trp Gly Asp Glu 35 40 45 Gln Ser Asp Phe Val Cys Asn Thr Gln Gln Pro Gly Cys Glu Asn Val 50 55 60 Cys Tyr Asp Glu Ala Phe Pro Ile Ser His Ile Arg Leu Trp Val Leu 65 70 75 80 Gln Ile Ile Phe Val Ser Thr Pro Ser Leu Met Tyr Val Gly His Ala 85 90 95 Val His Tyr Val Arg Met Glu Glu Lys Arg Lys Ser Arg Glu Ala Glu 100 105 110 Glu Leu Gly Gln Gln Ala Gly Thr Asn Gly Gly Pro Asp Gln Gly Ser 115 120 125 Val Lys Lys Ser Ser Gly Ser Lys Gly Thr Lys Lys Phe Arg Leu Glu 130 135 140 Gly Thr Leu Leu Arg Thr Tyr Ile Cys His Ile Ile Phe Lys Thr Leu 145 150 155 160 Phe Glu Val Gly Phe Ile Val Gly His Tyr Phe Leu Tyr Gly Phe Arg 165 170 175 Ile Leu Pro Leu Tyr Arg Cys Ser Arg Trp Pro Cys Pro Asn Val Val 180 185 190 Asp Cys Phe Val Ser Arg Pro Thr Glu Lys Thr Ile Phe Ile Leu Phe 195 200 205 Met Leu Ser Val Ala Ser Val Ser Leu Phe Leu Asn Val Met Glu Leu 210 215 220 Gly His Leu Gly Leu Lys Gly Ile Arg Ser Ala Leu Lys Arg Pro Val 225 230 235 240 Glu Gln Pro Leu Gly Glu Ile Pro Glu Lys Ser Leu His Ser Ile Ala 245 250 255 Val Ser Ser Ile Gln Lys Ala Lys Gly Tyr Gln Leu Leu Glu Glu Glu 260 265 270 Lys Ile Val Ser His Tyr Phe Pro Leu Thr Glu Val Gly Met Val Glu 275 280 285 Thr Ser Pro Leu Pro Ala Lys Pro Phe Asn Gln Phe Glu Glu Lys Ile 290 295 300 Ser Thr Gly Pro Leu Gly Asp Leu Ser Arg Gly Tyr Gln Glu Thr Leu 305 310 315 320 Pro Ser Tyr Ala Gln Val Gly Ala Gln Glu Val Glu Gly Glu Gly Pro 325 330 335 Pro Ala Glu Glu Gly Ala Glu Pro Glu Val Gly Glu Lys Lys Glu Glu 340 345 350 Ala Glu Arg Leu Thr Thr Glu Glu Gln Glu Lys Val Ala Val Pro Glu 355 360 365 Gly Glu Lys Val Glu Thr Pro Gly Val Asp Lys Glu Gly Glu Lys Glu 370 375 380 Glu Pro Gln Ser Glu Lys Val Ser Lys Gln Gly Leu Pro Ala Glu Lys 385 390 395 400 Thr Pro Ser Leu Cys Pro Glu Leu Thr Thr Asp Asp Ala Arg Pro Leu 405 410 415 Ser Arg Leu Ser Lys Ala Ser Ser Arg Ala Arg Ser Asp Asp Leu Thr 420 425 430 Val 20515PRThomo sapiensMISC_FEATURECx59 (NP_110399.2) 20Met Gly Asp Trp Asn Leu Leu Gly Asp Thr Leu Glu Glu Val His Ile 1 5 10 15 His Ser Thr Met Ile Gly Lys Ile Trp Leu Thr Ile Leu Phe Ile Phe 20 25 30 Arg Met Leu Val Leu Gly Val Ala Ala Glu Asp Val Trp Asn Asp Glu 35 40 45 Gln Ser Gly Phe Ile Cys Asn Thr Glu Gln Pro Gly Cys Arg Asn Val 50 55 60 Cys Tyr Asp Gln Ala Phe Pro Ile Ser Leu Ile Arg Tyr Trp Val Leu 65 70 75 80 Gln Val Ile Phe Val Ser Ser Pro Ser Leu Val Tyr Met Gly His Ala 85 90 95 Leu Tyr Arg Leu Arg Val Leu Glu Glu Glu Arg Gln Arg Met Lys Ala 100 105 110 Gln Leu Arg Val Glu Leu Glu Glu Val Glu Phe Glu Met Pro Arg Asp 115 120 125 Arg Arg Arg Leu Glu Gln Glu Leu Cys Gln Leu Glu Lys Arg Lys Leu 130 135 140 Asn Lys Ala Pro Leu Arg Gly Thr Leu Leu Cys Thr Tyr Val Ile His 145 150 155 160 Ile Phe Thr Arg Ser Val Val Glu Val Gly Phe Met Ile Gly Gln Tyr 165 170 175 Leu Leu Tyr Gly Phe His Leu Glu Pro Leu Phe Lys Cys His Gly His 180 185 190 Pro Cys Pro Asn Ile Ile Asp Cys Phe Val Ser Arg Pro Thr Glu Lys 195 200 205 Thr Ile Phe Leu Leu Phe Met Gln Ser Ile Ala Thr Ile Ser Leu Phe 210 215 220 Leu Asn Ile Leu Glu Ile Phe His Leu Gly Phe Lys Lys Ile Lys Arg 225 230 235 240 Gly Leu Trp Gly Lys Tyr Lys Leu Lys Lys Glu His Asn Glu Phe His 245 250 255 Ala Asn Lys Ala Lys Gln Asn Val Ala Lys Tyr Gln Ser Thr Ser Ala 260 265 270 Asn Ser Leu Lys Arg Leu Pro Ser Ala Pro Asp Tyr Asn Leu Leu Val 275 280 285 Glu Lys Gln Thr His Thr Ala Val Tyr Pro Ser Leu Asn Ser Ser Ser 290 295 300 Val Phe Gln Pro Asn Pro Asp Asn His Ser Val Asn Asp Glu Lys Cys 305 310 315 320 Ile Leu Asp Glu Gln Glu Thr Val Leu Ser Asn Glu Ile Ser Thr Leu 325 330 335 Ser Thr Ser Cys Ser His Phe Gln His Ile Ser Ser Asn Asn Asn Lys 340 345 350 Asp Thr His Lys Ile Phe Gly Lys Glu Leu Asn Gly Asn Gln Leu Met 355 360 365 Glu Lys Arg Glu Thr Glu Gly Lys Asp Ser Lys Arg Asn Tyr Tyr Ser 370 375 380 Arg Gly His Arg Ser Ile Pro Gly Val Ala Ile Asp Gly Glu Asn Asn 385 390 395 400 Met Arg Gln Ser Pro Gln Thr Val Phe Ser Leu Pro Ala Asn Cys Asp 405 410 415 Trp Lys Pro Arg Trp Leu Arg Ala Thr Trp Gly Ser Ser Thr Glu His 420 425 430 Glu Asn Arg Gly Ser Pro Pro Lys Gly Asn Leu Lys Gly Gln Phe Arg 435 440 445 Lys Gly Thr Val Arg Thr Leu Pro Pro Ser Gln Gly Asp Ser Gln Ser 450 455 460 Leu Asp Ile Pro Asn Thr Ala Asp Ser Leu Gly Gly Leu Ser Phe Glu 465 470 475 480 Pro Gly Leu Val Arg Thr Cys Asn Asn Pro Val Cys Pro Pro Asn His 485 490 495 Val Val Ser Leu Thr Asn Asn Leu Ile Gly Arg Arg Val Pro Thr Asp 500 505 510 Leu Gln Ile 515 21543PRThomo sapiensMISC_FEATURECx62 (NP_115991.1) 21Met Gly Asp Trp Asn Leu Leu Gly Gly Ile Leu Glu Glu Val His Ser 1 5 10 15 His Ser Thr Ile Val Gly Lys Ile Trp Leu Thr Ile Leu Phe Ile Phe 20 25 30 Arg Met Leu Val Leu Arg Val Ala Ala Glu Asp Val Trp Asp Asp Glu 35 40 45 Gln Ser Ala Phe Ala Cys Asn Thr Arg Gln Pro Gly Cys Asn Asn Ile 50 55 60 Cys Tyr Asp Asp Ala Phe Pro Ile Ser Leu Ile Arg Phe Trp Val Leu 65 70 75 80 Gln Ile Ile Phe Val Ser Ser Pro Ser Leu Val Tyr Met Gly His Ala 85 90 95 Leu Tyr Arg Leu Arg Ala Phe Glu Lys Asp Arg Gln Arg Lys Lys Ser 100 105 110 His Leu Arg Ala Gln Met Glu Asn Pro Asp Leu Asp Leu Glu Glu Gln 115 120 125 Gln Arg Ile Asp Arg Glu Leu Arg Arg Leu Glu Glu Gln Lys Arg Ile 130 135 140 His Lys Val Pro Leu Lys Gly Cys Leu Leu Arg Thr Tyr Val Leu His 145 150 155 160 Ile Leu Thr Arg Ser Val Leu Glu Val Gly Phe Met Ile Gly Gln Tyr 165 170 175 Ile Leu Tyr Gly Phe Gln Met His Pro Leu Tyr Lys Cys Thr Gln Pro 180 185 190 Pro Cys Pro Asn Ala Val Asp Cys Phe Val Ser Arg Pro Thr Glu Lys 195 200

205 Thr Ile Phe Met Leu Phe Met His Ser Ile Ala Ala Ile Ser Leu Leu 210 215 220 Leu Asn Ile Leu Glu Ile Phe His Leu Gly Ile Arg Lys Ile Met Arg 225 230 235 240 Thr Leu Tyr Lys Lys Ser Ser Ser Glu Gly Ile Glu Asp Glu Thr Gly 245 250 255 Pro Pro Phe His Leu Lys Lys Tyr Ser Val Ala Gln Gln Cys Met Ile 260 265 270 Cys Ser Ser Leu Pro Glu Arg Ile Ser Pro Leu Gln Ala Asn Asn Gln 275 280 285 Gln Gln Val Ile Arg Val Asn Val Pro Lys Ser Lys Thr Met Trp Gln 290 295 300 Ile Pro Gln Pro Arg Gln Leu Glu Val Asp Pro Ser Asn Gly Lys Lys 305 310 315 320 Asp Trp Ser Glu Lys Asp Gln His Ser Gly Gln Leu His Val His Ser 325 330 335 Pro Cys Pro Trp Ala Gly Ser Ala Gly Asn Gln His Leu Gly Gln Gln 340 345 350 Ser Asp His Ser Ser Phe Gly Leu Gln Asn Thr Met Ser Gln Ser Trp 355 360 365 Leu Gly Thr Thr Thr Ala Pro Arg Asn Cys Pro Ser Phe Ala Val Gly 370 375 380 Thr Trp Glu Gln Ser Gln Asp Pro Glu Pro Ser Gly Glu Pro Leu Thr 385 390 395 400 Asp Leu His Ser His Cys Arg Asp Ser Glu Gly Ser Met Arg Glu Ser 405 410 415 Gly Val Trp Ile Asp Arg Ser Arg Pro Gly Ser Arg Lys Ala Ser Phe 420 425 430 Leu Ser Arg Leu Leu Ser Glu Lys Arg His Leu His Ser Asp Ser Gly 435 440 445 Ser Ser Gly Ser Arg Asn Ser Ser Cys Leu Asp Phe Pro His Trp Glu 450 455 460 Asn Ser Pro Ser Pro Leu Pro Ser Val Thr Gly His Arg Thr Ser Met 465 470 475 480 Val Arg Gln Ala Ala Leu Pro Ile Met Glu Leu Ser Gln Glu Leu Phe 485 490 495 His Ser Gly Cys Phe Leu Phe Pro Phe Phe Leu Pro Gly Val Cys Met 500 505 510 Tyr Val Cys Val Asp Arg Glu Ala Asp Gly Gly Gly Asp Tyr Leu Trp 515 520 525 Arg Asp Lys Ile Ile His Ser Ile His Ser Val Lys Phe Asn Ser 530 535 540

Claims

1.-14. (canceled)

15. A method for potentiating the effects of a psychotropic drug in a patient in need thereof, said method comprising administration of an anti-connexin agent flecainide in said patient.

16. The method of claim 15, wherein said psychotropic drug is selected from the group consisting of: GABAergic, dopaminergic, norepinephrinergic, serotoninergic, histaminergic, and glutamatergic effectors, and those having an effect on the hypocretin/orexin system.

17. The method of claim 15, wherein said psychotropic drug is selected from the group consisting of: caffeine, mazindol, sodium oxybate, pitolisant, amphetamine, methylphenidate, (R)-(beta-amino-benzenepropyl) carbamate mono-hydrochloride, and armodafinil.

18. The method of claim 15, for potentiating the effects of a psychotropic drug selected from the group consisting of: caffeine, mazindol, sodium oxybate, pitolisant, amphetamine, methylphenidate, (R)-(beta-amino-benzenepropyl) carbamate mono-hydrochloride, and armodafinil.

19. The method of claim 15, wherein said anti-connexin agent Flecainide potentiates a promnesiant effect and/or an awakening effect of a psychotropic drug selected from the group consisting of: caffeine, mazindol, sodium oxybate, pitolisant, amphetamine, methylphenidate, (R)-(beta-amino-benzenepropyl) carbamate mono-hydrochloride, and armodafinil.

20. The method of claim 15, for increasing the efficacy and/or safety and/or the duration of action of said psychotropic drug.

21. The method of claim 15, wherein said flecainide is the R enantiomer of formula: ##STR00001##

22. A therapeutic composition comprising flecainide and at least one psychotropic drug, wherein said psychotropic drug is not flupirtine.

23. The therapeutic composition according to claim 22, wherein said psychotropic drug is selected from the group consisting of: GABAergic, dopaminergic, norepinephrinergic, serotoninergic, histaminergic, and glutamatergic effectors, and those having an effect on the hypocretin/orexin system.

24. The therapeutic composition according to claim 22, wherein said psychotropic drug is a psychotropic drug selected from the group consisting of: caffeine, mazindol, sodium oxybate, pitolisant, amphetamine, methylphenidate, (R)-(beta-amino-benzenepropyl) carbamate mono-hydrochloride, and armodafinil.

25. The therapeutic composition according to claim 22, wherein said flecainide is the R enantiomer of formula: ##STR00002##

26. A method for treating a disorder selected from the group consisting of: wounds, migraines, infectious diseases, drug-induced liver injury, cytotoxicity induced by chemotherapeutic agents, and ischemia, said method comprising administration of an anti-connexin agent flecainide in a patient in need thereof.

27. The method of claim 26, wherein said flecainide is the R enantiomer of formula: ##STR00003##