CYCLOBUTYL-UREA DERIVATIVES

Abstract

The invention relates to compounds of Formula (I)

##STR00001##

wherein X.sup.1, X.sup.2, X.sup.3, L, R.sup.X4, R.sup.1, R.sup.2A, R.sup.2B, R.sup.3, R.sup.4, R.sup.5, and R.sup.6 are as described in the description; to their preparation, to pharmaceutically acceptable salts thereof, to pharmaceutical compositions containing one or more compounds of Formula (I), and to the use of such compounds as medicaments, especially as Kv7 openers.

Claims

1. A compound of Formula (I) ##STR00047## wherein X.sup.1 represents nitrogen or CR.sup.X1; wherein R.sup.X1 represents hydrogen, halogen, (C.sub.1-4)alkyl, or (C.sub.1-4)alkoxy; X.sup.2 represents nitrogen or CR.sup.X2; wherein R.sup.X2 represents hydrogen, halogen, (C.sub.1-4)alkyl, or (C.sub.1-4)alkoxy; X.sup.3 represents nitrogen or CR.sup.X3; wherein R.sup.X3 represents hydrogen, halogen, (C.sub.1-4)alkyl, (C.sub.1-4)alkoxy, or hydroxy; R.sup.1 represents hydrogen or methyl; R.sup.X4 represents hydrogen, halogen, or (C.sub.1-4)alkyl; R.sup.2A represents hydrogen; (C.sub.1-4)alkyl; (C.sub.2-4)alkenyl; (C.sub.2-4)alkynyl; (C.sub.3-6)cycloalkyl; (C.sub.1- .sub.4)fluoroalkyl; (C.sub.1-4)hydroxyalkyl; (C.sub.1-4)alkoxy-(C.sub.1-2)alkyl; (C.sub.1-2)alkoxy-(C.sub.1-2)alkoxy-(C.sub.1- .sub.2)alkyl; (C.sub.1-2)alkyl-S-(C.sub.1-2)alkyl; (C.sub.1-2)alkyl-(SO.sub.2)-(C.sub.1-2)alkyl; cyano; (C.sub.1-2)cyanoalkyl; H.sub.2N-C(O)-(C.sub.1-2)alkyl; (R.sup.N1).sub.2N-(C.sub.1-2)alkyl or (R.sup.N1).sub.2N—C(O)—, wherein R.sup.N1 independently represents hydrogen or (C.sub.1-2)alkyl; or a 5-membered heteroaryl group containing one to four nitrogen atoms, wherein said 5-membered heteroaryl group is independently unsubstituted or mono-substituted with (C.sub.1-4)alkyl; and R.sup.2B represents hydrogen or methyl; or R.sup.2A and R.sup.2B form, together with the carbon atom to which they are attached, a ring of 3- to 6 members, wherein the members needed to complete said ring are each independently selected from —CH.sub.2— and —O— and wherein said ring does not contain more than one —O— member; L represents a direct bond, cycloprop-1,1-diyl, —CHR.sup.L—O—*, —O—CH.sub.2—*, —CH.sub.2—NH—*, —CH.sub.2—N(CH.sub.3)—*, —O—, or —(SO.sub.2)—; wherein R.sup.L represents hydrogen, (C.sub.1-4)alkyl, CH.sub.3—O—CH.sub.2—, or (CH.sub.3).sub.2NCH.sub.2-; wherein the asterisks indicate the bond which is linked to the aromatic carbon atom; R.sup.3 represents hydrogen or fluoro; R.sup.4 represents hydrogen or (C.sub.1-4)alkyl; R.sup.5 represents hydrogen, fluoro, or hydroxy; and R.sup.6 represents fluoro or (C.sub.1)fluoroalkyl; or R.sup.4 and R.sup.5 together represent a bridge selected from —CH.sub.2— and —CH.sub.2CH.sub.2—; and R.sup.6 represents hydrogen, fluoro, (C.sub.1)fluoroalkyl, or (C.sub.1-4)alkyl; or a salt thereof.

2. A compound according to claim 1, wherein X.sup.1 represents CR.sup.X1; wherein R.sup.X1 represents hydrogen or halogen; X.sup.2 represents nitrogen or CH; X.sup.3 represents nitrogen or CH; R.sup.1 represents hydrogen; R.sup.X4 represents hydrogen, halogen, or (C.sub.1-4)alkyl; R.sup.2A represents hydrogen; (C.sub.1-4)alkyl; (C.sub.1-4)fluoroalkyl; (C.sub.1-4)hydroxyalkyl; or (C.sub.1-4)alkoxy-(C.sub.1-2)alkyl; R.sup.2B represents hydrogen; L represents a direct bond, —CH.sub.2—O—*, or—O—; wherein the asterisk indicates the bond which is linked to the aromatic carbon atom; R.sup.3 represents hydrogen or fluoro; R.sup.4 represents hydrogen or (C.sub.1-4)alkyl; R.sup.5 represents hydrogen, fluoro, or hydroxy; and R.sup.6 represents fluoro or (C.sub.1)fluoroalkyl; or R.sup.4 and R.sup.5 together represent a bridge selected from —CH.sub.2— and —CH.sub.2CH.sub.2—; and R.sup.6 represents hydrogen, fluoro, (C.sub.1)fluoroalkyl, or (C.sub.1-4)alkyl; or a salt thereof.

3. A compound according to claim 2, wherein R.sup.2A represents hydrogen, (C.sub.1-4)alkyl, (C.sub.1-4)fluoroalkyl, (C.sub.1-4)hydroxyalkyl, or methoxymethyl; and R.sup.2B represents hydrogen; or a salt thereof.

4. A compound according to claim 3, wherein L represents a direct bond; or a salt thereof.

5. A compound according to claim 4, wherein R.sup.3 represents fluoro; or a salt thereof.

6. A compound according to claim 5, wherein R.sup.X4 represents hydrogen; or a salt thereof.

7. A compound according to claim 6, wherein each of X.sup.1, X.sup.2, and X.sup.3 represents CH; or a salt thereof.

8. A compound according to claim 3, wherein the fragment ##STR00048## represents: ##STR00049## wherein R.sup.X4 represents hydrogen or halogen; R.sup.3 represents hydrogen or fluoro; and L represents a direct bond, —CH.sub.2—O—*, or—O—; wherein the asterisk indicates the bond which is linked to the aromatic carbon atom; or ##STR00050## wherein X.sup.3 represents nitrogen or CH; R.sup.X4 represents hydrogen or (C.sub.1-4)alkyl; R.sup.3 represents hydrogen or fluoro; and L represents —CH.sub.2—O—*, or—O—; wherein the asterisk indicates the bond which is linked to the aromatic carbon atom; or a salt thereof.

9. A compound according to claim 7, wherein R.sup.4 represents hydrogen; R.sup.5 represents hydrogen or fluoro; and R.sup.6 represents fluoro, difluoromethyl or trifluoromethyl; or a salt thereof.

10. A compound according to claim 7, wherein R.sup.4 and R.sup.5 together represent a —CH.sub.2— bridge; and R.sup.6 represents hydrogen, fluoro, difluoromethyl, or trifluoromethyl; or a salt thereof.

11. A compound according to claim 1 selected from the group consisting of: 1-(3,3-Difluoro-cyclobutyl)-3-(3-trifluoromethyl-benzyl)-urea; 1-Bicyclo[1.1.1]pent-1-yl-3-[1-(3-trifluoromethyl-phenyl)-ethyl]-urea; 1-(3-Difluoromethyl-cyclobutyl)-3-[1-(3-trifluoromethyl-phenyl)-ethyl]-urea; 1-(3-Fluoro-bicyclo[1.1.1]pent-1-yl)-3-[1-(3-trifluoromethyl-phenyl)-ethyl]-urea; 1-(3-Difluoromethyl-bicyclo[1.1.1]pent-1-yl)-3-[1-(3-trifluoromethyl-phenyl)-ethyl]-urea; 1-Bicyclo[1.1.1]pent-1-yl-3-[1-(3-trifluoromethoxy-phenyl)-ethyl]-urea; 1-(3-Difluoromethyl-cyclobutyl)-3-[1-(3-trifluoromethoxy-phenyl)-ethyl]-urea; 1-(3-Fluoro-bicyclo[1.1.1]pent-1-yl)-3-[1-(3-trifluoromethoxy-phenyl)-ethyl]-urea; 1-[2,2-Difluoro-1-(3-trifluoromethyl-phenyl)-ethyl]-3-(3-hydroxy-3-trifluoromethyl-cyclobutyl)-urea; 1-(3,3-Difluoro-1-methyl-cyclobutyl)-3-[2,2-difluoro-1-(3-trifluoromethyl-phenyl)-ethyl]-urea; 1-Bicyclo[1.1.1]pent-1-yl-3-[1-(3-difluoromethoxy-phenyl)-ethyl]-urea; 1-(3-Difluoromethyl-bicyclo[1.1.1]pent-1-yl)-3-[2-hydroxy-1-(3-trifluoromethyl-phenyl)-ethyl]-urea; 1-(3-Difluoromethyl-bicyclo[1.1.1]pent-1-yl)-3-[2,2-difluoro-1-(3-trifluoromethyl-phenyl)-ethyl]-urea; 1-(3-Difluoromethyl-bicyclo[1.1.1]pent-1-yl)-3-[2-methoxy-1-(3-trifluoromethyl-phenyl)-ethyl]-urea; 1-(3-Difluoromethyl-bicyclo[1.1.1]pent-1-yl)-3-(2-fluoro-3-trifluoromethyl-benzyl)-urea; 1-(3-Difluoromethyl-bicyclo[1.1.1]pent-1-yl)-3-(3-fluoro-5-trifluoromethyl-benzyl)-urea; 1-(3-Fluoro-bicyclo[1.1.1]pent-1-yl)-3-[3-(2,2,2-trifluoro-ethoxy)-benzyl]-urea; 1-(3-Difluoromethyl-cyclobutyl)-3-[3-(2,2,2-trifluoro-ethoxy)-benzyl]-urea; 1-[3-(2,2,2-Trifluoro-ethoxy)-benzyl]-3-(3-trifluoromethyl-bicyclo[1.1.1]pent-1-yl)-urea; 1-(3-Difluoromethoxy-benzyl)-3-(3-fluoro-bicyclo[1.1.1]pent-1-yl)-urea; 1-(3-Difluoromethoxy-benzyl)-3-(3-difluoromethyl-cyclobutyl)-urea; 1-(3-Difluoromethoxy-benzyl)-3-(3-trifluoromethyl-bicyclo[1.1.1]pent-1-yl)-urea; 1-(3-Trifluoromethoxy-benzyl)-3-(3-trifluoromethyl-bicyclo[1.1.1]pent-1-yl)-urea; 1-(3-Fluoro-bicyclo[1.1.1]pent-1-yl)-3-(3-trifluoromethoxy-benzyl)-urea; 1-(3-Difluoromethyl-cyclobutyl)-3-(3-trifluoromethoxy-benzyl)-urea; 1-Bicyclo[1.1.1]pent-1-yl-3-(3-trifluoromethoxy-benzyl)-urea; 1-(3-Difluoromethyl-benzyl)-3-(3-difluoromethyl-cyclobutyl)-urea; 1-(3-Difluoromethyl-benzyl)-3-(3-trifluoromethyl-bicyclo[1.1.1]pent-1-yl)-urea; 1-(3-Difluoromethyl-cyclobutyl)-3-(2-trifluoromethoxy-pyridin-4-ylmethyl)-urea; 1-(2-Trifluoromethoxy-pyridin-4-ylmethyl)-3-(3-trifluoromethyl-bicyclo[1.1.1]pent-1-yl)-urea; 1-(3-Difluoromethyl-cyclobutyl)-3-{2-methoxy-1-[2-(2,2,2-trifluoro-ethoxy)-pyridin-4-yl]-ethyl}-urea; 1-{2-Methoxy-1-[2-(2,2, 2-trifluoro-ethoxy)-pyrid in-4-yl]-ethyl}-3-(3-trifluorom ethyl-cyclobutyl)-urea; 1-[1-(2-Difluoromethoxy-pyridin-4-yl)-ethyl]-3-(3-difluoromethyl-cyclobutyl)-urea; 1-{1-[2-Methyl-6-(2,2,2-trifluoro-ethoxy)-pyrimidin-4-yl]-ethyl}-3-(3-trifluoromethyl-bicyclo[1.1.1]pent-1-yl)-urea; 1-Bicyclo[1.1.1]pent-1-yl-3-(3-trifluoromethyl-benzyl)-urea; 1-(3-Fluoro-bicyclo[1.1.1]pent-1-yl)-3-(3-trifluoromethyl-benzyl)-urea; 1-(3-Trifluoromethyl-benzyl)-3-(3-trifluoromethyl-bicyclo[1.1.1]pent-1-yl)-urea; 1-(3-Difluoromethyl-cyclobutyl)-3-(3-trifluoromethyl-benzyl)-urea; 1-(3-Methyl-bicyclo[1.1.1]pent-1-yl)-3-(3-trifluoromethyl-benzyl)-urea; 1-(3-Fluoromethyl-bicyclo[1.1.1]pent-1-yl)-3-(3-trifluoromethyl-benzyl)-urea; 1-(3-Trifluoromethyl-benzyl)-3-(3-trifluoromethyl-cyclobutyl)-urea; 1-(3-Hydroxy-3-trifluoromethyl-cyclobutyl)-3-(3-trifluoromethyl-benzyl)-urea; 1-Bicyclo[1.1.1]pent-1-yl-3-[2-(2,2,2-trifluoro-ethoxy)-pyridin-4-ylmethyl]-urea; 1-(3-Fluoro-bicyclo[1.1.1]pent-1-yl)-3-[2-(2,2,2-trifluoro-ethoxy)-pyridin-4-ylmethyl]-urea; 1-[2-(2,2,2-Trifluoro-ethoxy)-pyridin-4-ylmethyl]-3-(3-trifluoromethyl-bicyclo[1.1.1]pent-1-yl)-urea; 1-(3-Difluoromethyl-cyclobutyl)-3-[2-(2,2,2-trifluoro-ethoxy)-pyridin-4-ylmethyl]-urea; 1-(3-Difluoromethyl-bicyclo[1.1.1]pent-1-yl)-3-(3-trifluoromethyl-benzyl)-urea; 1-Bicyclo[2.1.1]hex-1-yl-3-(3-trifluoromethyl-benzyl)-urea; 1-(3,3-Difluoro-1-methyl-cyclobutyl)-3-(3-trifluoromethyl-benzyl)-urea; 1-(3-(trifluoromethyl)benzyl)-3-((1s,3s)-3-(trifluoromethyl)cyclobutyl)urea; 1-(3-(trifluoromethyl)benzyl)-3-((1r,3r)-3-(trifluoromethyl)cyclobutyl)urea; 1-((1s,3s)-3-(difluoromethyl)cyclobutyl)-3-(3-(trifluoromethyl)benzyl)urea; 1-((1r,3r)-3-(difluoromethyl)cyclobutyl)-3-(3-(trifluoromethyl)benzyl)urea; 1-{(S)-1-[2-Methyl-6-(2,2,2-trifluoro-ethoxy)-pyrimidin-4-yl]-ethyl}-3-(3-trifluoromethyl-bicyclo[1.1.1]pent-1-yl)-urea; 1-((1r,3r)-3-(difluoromethyl)cyclobutyl)-3-((2-(2,2,2-trifluoroethoxy)pyridin-4-yl)methyl)urea; 1-((1s,3s)-3-(difluoromethyl)cyclobutyl)-3-((2-(2,2,2-trifluoroethoxy)pyridin-4-yl)methyl)urea; 1-((1s,3R)-3-(difluoromethyl)cyclobutyl)-3-((S)-1-(3-(trifluoromethoxy)phenyl)ethyl)urea; and 1-((1r,3S)-3-(difluoromethyl)cyclobutyl)-3-((S)-1-(3-(trifluoromethoxy)phenyl)ethyl)urea; or a salt thereof.

12. A pharmaceutical composition comprising, as active principle, a compound according to claim 1, or a pharmaceutically acceptable salt thereof, and at least one therapeutically inert excipient.

13. (canceled)

14. A method for prevention or treatment of a disease selected from epilepsy, myokymia, tinnitus, hearing disorders, neuropathic and inflammatory pain, psychiatric disorders, substance use disorders, neurological disorders, and diseases affecting the smooth muscles in a patient, wherein the method comprises administering to the patient a pharmaceutically active amount of a compound according to claim 1 or a pharmaceutically acceptable salt thereof.

15. (canceled)

16. A compound according to claim 8, wherein R.sup.4 represents hydrogen; R.sup.5 represents hydrogen or fluoro; and R.sup.6 represents fluoro, difluoromethyl or trifluoromethyl; or a salt thereof.

17. A compound according to claim 8, wherein R.sup.4 and R.sup.5 together represent a —CH.sub.2— bridge; and R.sup.6 represents hydrogen, fluoro, difluoromethyl, or trifluoromethyl; or a salt thereof.

18. A compound according to claim 7, wherein R.sup.1 represents hydrogen; or a salt thereof.

19. A compound according to claim 9, wherein R.sup.1 represents hydrogen; or a salt thereof.

20. A compound according to claim 10, wherein R.sup.1 represents hydrogen; or a salt thereof.

21. A compound according to claim 1, wherein the compound is 1-(3-difluoromethyl-bicyclo[1.1.1]pent-1-yl)-3-(3-trifluoromethyl-benzyl)-urea.

22. A compound according to claim 1, wherein the compound is 1-(3-(difluoromethyl)cyclobutyl)-3-(3-(trifluoromethyl)benzyl)urea.

23. A pharmaceutical composition comprising, as active principle, the compound according to claim 21 and at least one therapeutically inert excipient.

24. A pharmaceutical composition comprising, as active principle, the compound according to claim 22 and at least one therapeutically inert excipient.

25. A method for prevention or treatment of a disease selected from epilepsy, myokymia, tinnitus, hearing disorders, neuropathic and inflammatory pain, psychiatric disorders, substance use disorders, neurological disorders, and diseases affecting the smooth muscles in a patient, wherein the method comprises administering to the patient a pharmaceutically active amount of the compound according to claim 21.

26. A method for prevention or treatment of a disease selected from epilepsy, myokymia, tinnitus, hearing disorders, neuropathic and inflammatory pain, psychiatric disorders, substance use disorders, neurological disorders, and diseases affecting the smooth muscles in a patient, wherein the method comprises administering to the patient a pharmaceutically active amount of the compound according to claim 22.

Description

PREPARATION OF COMPOUNDS OF FORMULA (L)

[0208] A further aspect of the invention is a process for the preparation of compounds of Formula (I). Compounds according to Formula (l) of the present invention can be prepared from commercially available or well known starting materials according to the methods described in the experimental part; by analogous methods; or according to the general sequence of reactions outlined below, wherein R.sup.1, R.sup.2A, R.sup.2B, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.X4, X.sup.1, X.sup.2, X.sup.3, and L are as defined for Formula (l). Other abbreviations used herein are explicitly defined, or are as defined in the experimental section. In some instances the generic groups R.sup.1, R.sup.2A, R.sup.2B, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.X4, X.sup.1, X.sup.2, X.sup.3, and L might be incompatible with the assembly illustrated in the schemes below and so will require the use of protecting groups (PG). The use of protecting groups is well known in the art (see for example “Protective Groups in Organic Synthesis”, T.W. Greene, P.G.M. Wuts, Wiley-Interscience, 1999). For the purposes of this discussion, it will be assumed that such protecting groups as necessary are in place. The compounds obtained may also be converted into salts, especially pharmaceutically acceptable salts thereof in a manner known per se.

General Preparation Routes

[0209] Generally, compounds of Formula l can be synthesised by treating an amine of Stucture 2 (or the corresponding salt, like HCl or TFA salts) with an isocyanate 3 in the presence of a base such as NEt.sub.3 or DIPEA in solvent such as DCM or MeCN. Alternatively, an isocyanate of Structure 4 can be reacted with an amine 5 (or the corresponding salt, like HCl or TFA salts) in the presence of a base such as NEt.sub.3 or DIPEA in solvent such as DCM or MeCN to afford compounds of Formula I-A (Scheme 1).

##STR00039##

[0210] Alternatively, an amine of Structure 2 (or the corresponding salt, like HCl or TFA salts) is condensed with 4-nitrophenyl chloroformate in the presence of a base like NEt.sub.3 or DIPEA to give a carbamate 6 (Scheme 2). The carbamate 6 is then treated with an amine 5 (or the corresponding salt, like HCl or TFA salts) in the presence of a base like NEt.sub.3 in a solvent like THF to yield a compound of Formula l. The sequence can also start by first reacting an amine 5 (or the corresponding salt, like HCl or TFA salts) with 4-nitrophenyl chloroformate in the presence of a base like NEt.sub.3 or DIPEA to give a carbamate 7 (Scheme 2). The carbamate 7 is then treated with an amine of Structure 2 (or the corresponding salt, like HCl or TFA salts) in the presence of a base like NEt.sub.3 in solvent like THF to yield a compound of Formula l.

##STR00040##

[0211] In another aspect, an amine of Structure 2 (or the corresponding salt, like HCl or TFA salts) is activated with a reagent like CDI, triphosgene, or trifluoroethoxycarbonate and the activated intermediate is in-situ treated with an amine 5 (or the corresponding salt, like HCl or TFA salts) to yield a compound of Formula l (Scheme 3). Conversely, an amine 5 (or the corresponding salt, like HCl or TFA salts) can be activated with a reagent like CDI, triphosgene, or trifluoroethoxycarbonate and the activated intermediate is in-situ treated with an amine of Structure 2 (or the corresponding salt, like HCl or TFA salts) to yield a compound of Formula l.

##STR00041##

[0212] Amines of Structure 2-A or 2-B can be synthesized by taking advantage of the Ellman’s auxiliary (Scheme 4). Thus, an aldehyde 8 is treated with tert-butanesulfinamide 9 in the presence of Ti(OEt).sub.4 to provide a tert-butanesulfinyl imine 10. Compound 10 is then treated with a nucleophile such as a Grignard reagent 11 to afford a protected amine 12. The tert-butanesulfinyl group is then cleaved under mild acidic conditions like HCl in MeOH to afford an amine of Structure 2-A (or the corresponding HCl salt). Alternatively, imine 10 can be reduced with a reducing agent like NaBH.sub.4 in MeOH to yield a protected amine 13. The tert-butanesulfinyl group is then cleaved under mild acidic conditions like HCl in MeOH to afford an amine of Structure 2-B (or the corresponding HCl salt). Alternatively, a ketone 14 can be reacted with tert-butanesulfinamide 9 in the presence of Ti(OEt).sub.4 to provide a tert-butanesulfinyl imine 15. Compound 15 is then treated with a Grignard or lithiated reagent 16 to afford a protected amine 17. The tert-butanesulfinyl group is then cleaved under mild acidic conditions like HCl in MeOH to afford an amine of Structure 2-C (or the corresponding HCl salt).

[0213] In another aspect, an amine of Structure 2-A can be synthesized using photoredox catalysis (Scheme 5). A bromide 18 is reacted with a Boc-protected amino acid 19 in the presence of an iridium catalyst like [lr{dF(CF.sub.3)ppy}.sub.2(dtbpy)]PF.sub.6 and a nickel catalyst like NiCl.sub.2•glyme in a solvent like DMSO or DMA under blue LED irradiation to give a Boc-protected amine 20 (Science 2014, 345, 437-440). The Boc-protecting group is then cleaved under acidic conditions like TFA in DCM or 4 M HCl in dioxane to give an amine of Structure 2-A (or the corresponding salt, like HCl or TFA salts).

##STR00042##

[0214] An amine of Structure 2-B can also be obtained from the corresponding nitrile 21 (Scheme 6). A solution of a nitrile 21 in MeOH can be reduced using a catalyst like Ra/Ni under an H.sub.2-atmosphere (in flow or batch mode) or LiAlH.sub.4 in a solvent like THF to give an amine of Structure 2-B. Alternatively, nitrile 21 can be reduced using a nickel catalyst like NiCI.sub.2•6H.sub.2O and NaBH.sub.4 in the presence of Boc.sub.2O to give a Boc-protected amine 22. Deprotection under acidic conditions like TFA in DCM or HCl in dioxane yield an amine of Structure 2-B (or the corresponding HCl or TFA salt). Nitrile 21 can also be converted to the corresponding ketones 23 using MeMgBr in a solvent like Et.sub.2O followed by an aqueous acidic treatment. Ketone 23 can undergo a reductive amination in a solvent like MeOH with for example ammonium acetate and sodium cyanoborohydride to give an amine of Structure 2-A (where R.sup.2A is methyl). Moreover, nitrile 21 can be treated first with MeMgBr in a solvent like 2-methyltetrahydrofuran and then with NaBH.sub.4 to give an amine of Structure 2-A (where R.sup.2A is methyl). Nitrile 21 can also be subjected to a Kulinkovich reaction in Et.sub.2O using EtMgBr in the presence of a titanium salt like Ti(OiPr).sub.4 and borontrifluoride to give an amine of Structure 2-D. Finally, nitrile 21 can react with a Boc-protected amino acid in the presence of cesium fluoride and an iridium catalyst like Ir(p-F(t-Bu)-ppy).sub.3 in a solvent like DMSO or DMA under blue LED irradiation to give a Boc-protected amine 24 (JACS 2014, 136, 5257-5260). The protecting group can then be cleaved under acidic conditions like TFA in DCM or HCl in dioxane to give an amine of Structure 2-E (or the corresponding HCl or TFA salt).

##STR00043##

[0215] Aldehydes 8-A can be prepared as described in Scheme 7. Thus, alcohol 25 can be reacted with an alkylating agent like alkylsulfonate, alkylbromide, or alkyliodide in the presence of a base like Cs.sub.2CO.sub.3 or K.sub.2CO.sub.3 in a solvent like DMF to give an aldehyde 8-A. Similarly, an alcohol 26 can be converted into the corresponding bromide 18-A.

##STR00044##

[0216] Nitriles 21-A are obtained through a S.sub.NAr reaction between a chloro or fluoro nitrile 27 and an alcohol like trifluoroethanol in the presence of a base like sodium hydride in a solvent like THF (Scheme 8). Alternatively, nitrile 27 can undergo a S.sub.NAr reaction with an amine (or the corresponding HCl salt) in a solvent like NMP and a base like NEt.sub.3 under microwave irradiation to yield a nitrile 21-B. Finally, a cyanation between chloro or bromo derivative 28 and ZnCN.sub.2 in the presence of a palladium catalyst like Pd.sub.2(dba).sub.3 and a ligand like dppf in a solvent like DMF give a nitrile 21.

##STR00045##

[0217] An amine of Structure 2 can also be prepared by methods described in Scheme 9. Thus, a Boc-protected amine 30 can be treated with an alkylating agent like alkyl bromide or alkyl iodide in the presence of a base or a silver salt like Ag.sub.2O to give a Boc-protected amine 31. The Boc-protecting group is then cleaved under acidic conditions like TFA in DCM or 4 M HCl in dioxane to give an amine of Structure 2 (or the corresponding salt, like HCl or TFA salt). Alternatively, an aldehyde 8 can undergo a reductive amination with an amine 32 in a solvent like DCM and in the presence of a reducing agent like NaBH(OAc).sub.3 and a base like DIPEA to give an amine of Structure 2, wherein R.sup.2A and R.sup.2B represent hydrogen.

##STR00046##

Experimental Section

[0218] TABLE-US-00001 Abbrevations (as used herein and in the description above) anh. anhydrous Ac acetyl aq. aqueous Boc tert.-butyloxycarbonyl Bu butyl CDI 1,1′-carbonyldiimidazole comb. Combined d day(s) dba dibenzylideneacetone DCM dichloromethane DIPEA N-ethyldiisopropylamine DMA dimethylacetamide DMF dimethylformamide DMSO dimethylsulfoxide dppf 1,1′-ferrocenediyl-bis(diphenylphosphine) eq equivalent Et ethyl FBS fetal bovine serum FLIPR Fluorescent imaging plate reader Fluo-8-AM h bis(acetoxymethyl) 2,2′-((4-(6-(acetoxymethoxy)-3-oxo-3H-xanthen-9-yl)-2-(2-(bis(2-acetoxymethoxy)-2-oxoethyl)amino)phenoxy)ethoxy)phenyl)azanediyl)diacetate hour(s) HATU 2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate HBSS Hank’s balanced salt solution HEK293 Human embryonic kidney 293 cells HEPES 4-(2-hydroxyethyl)-piperazine-1-ethanesulfonic acid Hept heptane(s) HV High vacuum HPLC high performance liquid chromatography iPr isopropyl lr(p-F(t-Bu)-ppy).sub.3 tris (2- (3-tert-butylphenyl) -4-tert-butylpyridine) iridium [lr{dF(CF.sub.3)ppy}.sub.2(dtbpy)]PF.sub.6 [4,4′-Bis(1,1-dimethylethyl)-2,2′-bipyridine-N1,N1′]bis[3,5-difluoro-2-[5-(trifluoromethyl)-2-pyridinyl-N] phenyl-C]lridium(III) hexafluorophosphate LC liquid chromatography LED light-emitting diode M molarity [mol L.sup.-1] Me methyl MS mass spectroscopy min minute(s) N normality NiCl.sub.2•glyme Nickel(II) chloride ethylene glycol dimethyl ether complex NMDA N-methyl-D-aspartate NMP N-methyl-2-pyrrolidone NMR Nuclear magnetic resonance org. organic PBS phosphate-buffered saline PG protecting group Ph phenyl Prep. Preparative Ra/Ni Raney-Nickel rpm rotations per minute rt room temperature sat. saturated sec second(s) SFC supercritical fluid chromatography soln. solution TBME tert-butyl methyl ether tBu tert-butyl TFA trifluoroacetic acid THF tetrahydrofuran t.sub.R retention time UPLC Ultra performance liquid chromatography UV ultraviolet XE-991 10,10-bis(4-pyridinylmethyl)-9(10H)-anthracenone

I. Chemistry

[0219] The following examples illustrate the preparation of biologically active compounds of the invention but do not at all limit the scope thereof.

[0220] General remarks: All solvents and reagents are used as obtained from commercial sources unless otherwise indicated. Temperatures are indicated in degrees Celsius (°C). Unless otherwise indicated, the reactions take place at room temperature (rt) under an argon or nitrogen atmosphere and are run in a flame dried round-bottomed flask equipped with a magnetic stir bar. In mixtures, relations of parts of solvent or eluent or reagent mixtures in liquid form are given as volume relations (v/v), unless indicated otherwise.

Characterization Methods Used

[0221] LC-MS 1

[0222] LC-MS-conditions: Analytical. Pump: Waters Acquity Binary, Solvent Manager, MS: Waters SQ Detector or Xevo TQD, DAD: Acquity UPLC PDA Detector. Column: Acquity UPLC CSH C18 1.7 um, 2.1 × 50 mm from Waters, thermostated in the Acquity UPLC Column Manager at 60° C. Eluents: A1: H.sub.2O + 0.05% formic acid; B1: MeCN + 0.045% formic acid. Method: Gradient: 2% B to 98% B over 2.0 min. Flow: 1.0 mL/min. Detection at 214 nm and MS, retention time t.sub.R is given in min.

LC-MS 2 to 4

[0223] UPLC/MS analyses are performed on Acquity UPLC setup. The column temperature is 40° C.

[0224] The LC retention times are obtained using the following elution conditions: LC-MS 2: Analytical UPLC on a Agilent Zorbax RRHD SB-Aq (2.1×50 mm, 1.8 um); detection at 210 nM and MS; Gradient of water/ 0.04% TFA (A) and MeCN (B). The eluent flow rate was 0.8 mL/min and the characteristics of the eluting mixture proportion in function of the time t from start of the elution are summarized in the table below (a linear gradient being used between two consecutive time points):

TABLE-US-00002 t (min) 0 1.2 1.9 2.1 Solvent A (%) 95 5 5 95 Solvent B (%) 5 95 95 5

LC-MS 3: Analytical UPLC on a Waters Xbridge (4.6×30 mm, 2.5 um); detection at 210 nM and MS; Gradient of water/ 0.04% TFA (A) and MeCN (B). The eluent flow rate was 4.5 mL/min and the characteristics of the eluting mixture proportion in function of the time t from start of the elution are summarized in the table below (a linear gradient being used between two consecutive time points):

TABLE-US-00003 t (min) 0 1.00 1.45 1.55 Solvent A (%) 95 5 5 95 Solvent B (%) 5 95 95 5

LC-MS 4: Analytical UPLC on a Waters BEH C18 (2.1×50 mm, 2.5 um); detection at 210 nM and MS; Gradient of water/ 0.04% NH.sub.3 [c(NH.sub.3) = 13 mmol/l] (A) and MeCN (B). The eluent flow rate was 0.8 mL/min and the characteristics of the eluting mixture proportion in function of the time t from start of the elution are summarized in the table below (a linear gradient being used between two consecutive time points):

TABLE-US-00004 t (min) 0 1.2 1.9 2.1 Solvent A (%) 95 5 5 95 Solvent B (%) 5 95 95 5

Preparative LC-MS Methods Used

[0225] Preparative HPLC/MS purifications are performed on a Gilson HPLC system, equipped with a Gilson 215 autosampler, Gilson 333/334 pumps, Finnigan AQA MS detector system, and a Dionex UV detector, using a Waters Xbridge C18 or an Waters Atlantis column, with a linear gradient of water/formic acid 0.02% (A) and MeCN (B) (acidic conditions) or water/ammonia 0.02% (A) and MeCN (B) (basic conditions).

Combiflash

[0226] Flash column chromatography was performed using a combiflash from Teledyne ISCO.

Preparative Chiral SFC Methods Used

[0227] Preparative chiral SFC purifications are performed on a Sepiatec Prep SFC 360 system. Following parameters were used: [0228] Preparative chiral SFC 1: A ChiralPak IB column (30×250 mm, 5 um) was used. The modifier was iPrOH (12%), run for 5 min and at a flow rate of 160 mL/min. The following system settings were used: backpressure 100 bar, temperature pumphead 5° C., temperature fraction module 20° C., and temperature column department 40° C. [0229] Preparative chiral SFC 2: A ChiralPak IH column (30×250 mm, 5 um) was used. The modifier was EtOH (15%), run for 3.3 min and at a flow rate of 160 mL/min. The following system settings were used: backpressure 100 bar, temperature pumphead 5° C., temperature fraction module 20° C., and temperature column department 40° C. [0230] Preparative chiral SFC 3: A Regis (R,R)Whelk-O1column (30×250 mm, 5 um) was used. The modifier was EtOH (15%), run for 3.0 min and at a flow rate of 160 mL/min. The following system settings were used: backpressure 100 bar, temperature pumphead 5° C., temperature fraction module 20° C., and temperature column department 40° C. [0231] Preparative chiral SFC 4: A ChiralPak IB column (30×250 mm, 5 um) was used. The modifier was EtOH (10%), run for 5.5 min and at a flow rate of 160 mL/min. The following system settings were used: backpressure 100 bar, temperature pumphead 5° C., temperature fraction module 20° C., and temperature column department 40° C. [0232] Preparative chiral SFC 5: A Regis (R,R)Whelk-O1column (30×250 mm, 5 um) was used. The modifier was MeOH (20%), run for 4.0 min and at a flow rate of 160 mL/min. The following system settings were used: backpressure 100 bar, temperature pumphead 5° C., temperature fraction module 20° C., and temperature column department 40° C. [0233] Preparative chiral SFC 6: A ChiralPak AD-H column (30×250 mm, 5um) was used. The modifier was EtOH (10%), run for 3.0 min and at a flow rate of 160 mL/min. The following system settings were used: backpressure 100 bar, temperature pumphead 5° C., temperature fraction module 20° C., and temperature column department 40° C.

NMR

[0234] .sup.1H-NMR spectra were recorded at rt with a Brucker NMR 500 spectrometer 1H (500 MHz) equipped with a Bruker’s DCH cryoprobe. Chemical shifts are reported in ppm downfield from tetramethylsilane using residual solvent signals as internal reference. The multiplicity is described as singulet s, doublet d, triplet t, quadruplet q, hextet h, or multiplet m. Broad signals are indicated as br.

Example 1: 1-(3,3-Difluoro-Cyclobutyl)-3-(3-Trifluoromethyl-Benzyl)-Urea

[0235] To a solution of 3-(trifluoromethyl)benzylamine (18 mg, 0.1 mmol, 1.0 eq) in MeCN (0.5 mL), DIPEA (19 .Math.L, 0.11 mmol, 1.1 eq) and a solution of CDI (32 mg, 0.2 mmol, 2.0 eq) in MeCN (0.2 mL) were added in sequence. The mixture was stirred at 60° C. for 3 hours. A solution of 3,3-difluorocyclobutan-1-amine (21 mg, 0.2 mmol, 2.0 eq) in MeCN (0.5 mL) and H.sub.2O (0.1 mL) was added. The mixture was further stirred at 60° C. overnight. The mixture was allowed to cool to rt and purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 0.99 min; [M+H].sup.+: 309.2.

Example 2: 1-Bicyclo[1.1.1]Pent-1-yl-3-[1-(3-Trifluoromethyl-Phenyl)-Ethyl]-Urea

[0236] To a solution of bicyclo[1.1.1]pentan-1-amine hydrochloride(12 mg, 0.1 mmol, 1 eq) in MeCN (0.5 mL), DIPEA (34 .Math.L, 0.2 mmol, 2 eq) and CDI (16 mg, 0.1 mmol, 1 eq) were added in sequence. The mixture was stirred at 60° C. for 1 hour. A solution of 1-(3-trifluoromethylphenyl)ethylamine (19 mg, 0.1 mmol, 1 eq) in MeCN (0.4 mL) and H.sub.2O (0.1 mL) was added. The reaction mixture was further stirred at 60° C. overnight. The mixture was allowed to cool to rt and purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 1.08 min; [M+H].sup.+: 299.2.

[0237] Example 3 to Example 5 were synthesized using the appropriate amine or amine salt (HCI or TFA) derivative and following the procedure described in Example 2. LC-MS data of Example 3 to Example 5 are listed in the table below. The LC-MS conditions used were LC-MS (1).

TABLE-US-00005 Example N° Name t.sub.R [M+H].sup.+ 3 1-(3-Difluoromethyl-cyclobutyl)-3-[1-(3-trifluoromethylphenyl)-ethyl]-urea 1.05 337.3 4 1-(3-Fluoro-bicyclo[1.1.1]pent-1-yl)-3-[1-(3-trifluoromethylphenyl)-ethyl]-urea 1.07 317.2 5 1-(3-Difluoromethyl-bicyclo[1.1.1]pent-1-yl)-3-[1-(3-trifluoromethyl-phenyl)-ethyl]-urea 1.10 349.3

Example 6: 1-Bicyclo[1.1.1]Pent-1-yl-3-[1-(3-Trifluoromethoxy-Phenyl)-Ethyl]-Urea

[0238] To a solution of bicyclo[1.1.1]pentan-1-amine hydrochloride (12 mg, 0.1 mmol, 1 eq) in MeCN (0.5 mL), DIPEA (34 .Math.L, 0.2 mmol, 2 eq) and CDI (16 mg, 0.1 mmol, 1 eq) were added in sequence. The mixture was stirred at 60° C. for 1 hour. A solution of 1-(3-(trifluoromethoxy)phenyl)ethanamine (21 mg, 0.1 mmol, 1 eq) in MeCN (0.4 mL) and H.sub.2O (0.1 mL) was added. The reaction mixture was further stirred at 60° C. overnight. The mixture was allowed to cool to rt and purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 1.11 min; [M+H].sup.+: 315.2.

[0239] Example 7 to Example 8 were synthesized using the appropriate amine or amine salt (HCI or TFA) derivative and following the procedure described in Example 6. LC-MS data of Example 7 to Example 8 are listed in the table below. The LC-MS conditions used were LC-MS (1).

TABLE-US-00006 Example N° Name t.sub.R [M+H].sup.+ 7 1-(3-Difluoromethyl-cyclobutyl)-3-[1-(3-trifluoromethoxyphenyl)-ethyl]-urea 1.08 353.2 8 1-(3-Fluoro-bicyclo[1.1.1]pent-1-yl)-3-[1-(3-trifluoromethoxyphenyl)-ethyl]-urea 1.10 333.2

Example 9: 1-[2,2-Difluoro-1-(3-Trifluoromethyl-Phenyl)-Ethyl]-3-(3-Hydroxy-3-Trifluoromethyl-Cyclobutyl)-Urea

[0240] To a solution of 3-amino-1-(trifluoromethyl)cyclobutan-1-ol (19 mg, 0.12 mmol, 1.5 eq) in MeCN (0.1 mL), a solution of CDI (20 mg, 0.12 mmol, 1.5 eq) in MeCN (0.2 mL) was added. The reaction mixture was stirred at rt for 2 hours. A solution of 2,2-difluoro-1-[3-(trifluoromethyl)phenyl]ethan-1-amine (19 mg, 0.08 mmol, 1.0 eq) and DIPEA (15 .Math.L, 0.09 mmol, 1.1 eq) in MeCN (0.5 mL) and H.sub.2O (0.1 mL) was added. The mixture was stirred at rt overnight. The mixture was purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 1.01 min; [M+H].sup.+: 407.3.

Example 10: 1-(3,3-Difluoro-1-Methyl-Cyclobutyl)-3-[2,2-Difluoro-1-(3-TrifluoromethylPhenyl)-Ethyl]-Urea

[0241] The product was synthesized using 3-3-difluoro-1-methylcyclobutanamine-hydrochloride and following the procedure described in Example 9. LC-MS (1): t.sub.R = 1.13 min; [M+H].sup.+: 373.3.

Example 11: 1-Bicyclo[1.1.1]Pent-1-yl-3-[1-(3-Difluoromethoxy-Phenyl)-Ethyl]-Urea

[0242] To a solution of 1-(3-(difluoromethoxy)phenyl)ethan-1-amine hydrochloride (37 mg, 0.1 mmol, 1 eq) in MeCN (0.5 mL), DIPEA (51 .Math.L, 0.3 mmol, 3 eq) and CDI (32 mg, 0.2 mmol, 2 eq) were added in sequence. The reaction was stirred at rt for 1 h. A solution of bicyclo[1.1.1]pentan-1-amine hydrochloride in MeCN (0.4 mL) and H.sub.2O (0.1 mL) was added. The reaction mixture was stirred at rt for 1 hour. The mixture was purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 1.01 min; [M+H].sup.+: 297.3.

Example 12: 1-(3-Difluoromethyl-Bicyclo[1.1.1]Pent-1-yl)-3-[2-Hydroxy-1-(3-Trifluoromethyl-Phenyl)-Ethyl]-Urea

[0243] To a solution of 3-(difluoromethyl)bicyclo[1.1.1]pentan-1-amine hydrochloride (25 mg, 0.15 mmol, 1.0 eq) in MeCN (0.8 mL), DIPEA (92 .Math.L, 0.53 mmol, 3.5 eq) and CDI (37 mg, 0.23 mmol, 1.5 eq) were added in sequence. The mixture was stirred at 50° C. for 40 min. 2-Amino-2-(3-trifluoromethyl-phenyl)-ethanol (31 mg, 0.15 mmol, 1.0 eq) was added. The reaction mixture was stirred at 80° C. for 18 hours. The mixture was allowed to cool to rt and purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 0.96 min; [M+H].sup.+: 365.2.

[0244] Example 13 to Example 16 were synthesized using the appropriate amine or amine salt (HCI or TFA) derivative and following the procedure described in Example 12. LC-MS data of Example 13 to Example 16 are listed in the table below. The LC-MS conditions used were LC-MS (1).

TABLE-US-00007 Example N° Name t.sub.R [M+H].sup.+ 13 1-(3-Difluoromethyl-bicyclo[1.1.1]pent-1-yl)-3-[2,2-difluoro-1-(3-trifluoromethyl-phenyl)-ethyl]-urea 1.12 385.3 14 1-(3-Difluoromethyl-bicyclo[1.1.1]pent-1-yl)-3-[2-methoxy-1-(3-trifluoromethyl-phenyl)-ethyl]-urea 1.08 379.3 15 1-(3-Difluoromethyl-bicyclo[1.1.1]pent-1-yl)-3-(2-fluoro-3-trifluoromethyl-benzyl)-urea 1.07 353.2 16 1-(3-Difluoromethyl-bicyclo[1.1.1]pent-1-yl)-3-(3-fluoro-5-trifluoromethyl-benzyl)-urea 1.09 353.2

Example 17: 1-(3-Fluoro-Bicyclo[1.1.1]Pent-1-yl)-3-[3-(2,2,2-Trifluoro-Ethoxy)-Benzyl]-Urea

[0245] A solution of (3-(2,2,2-trifluoroethoxy)phenyl)methanamine (14 mg, 0.07 mmol, 1.0 eq) in MeCN (0.4 mL) was treated at rt with DIPEA (43 .Math.L, 0.25 mmol, 3.5 eq) followed by CDI (12 mg, 0.07 mmol, 1.05 eq) and the resulting mixture was stirred at 50° C. for 30 min. The resulting mixture was treated with 3-fluorobicyclo[1.1.1]pentan-1-amine hydrochloride (10 mg, 0.07 mmol, 1.0 eq) and the reaction mixture was stirred at 80° C. overnight. The mixture was allowed to cool to rt and purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 1.02 min; [M+H].sup.+: 333.3.

[0246] Example 18 to Example 19 were synthesized using the appropriate amine or amine salt (HCI or TFA) derivative and following the procedure described in Example 17. LC-MS data of Example 18 to Example 19 are listed in the table below. The LC-MS conditions used were LC-MS (1).

TABLE-US-00008 Example N° Name t.sub.R [M+H].sup.+ 18 1-(3-Difluoromethyl-cyclobutyl)-3-[3-(2,2,2-trifluoro-ethoxy)-benzyl]-urea 1.01 353.3 19 1-[3-(2,2,2-Trifluoro-ethoxy)-benzyl]-3-(3-trifluoromethyl-bicyclo[1.1.1]pent-1-yl)-urea 1.14 383.3

Example 20: 1-(3-Difluoromethoxy-Benzyl)-3-(3-Fluoro-Bicyclo[1.1.1]Pent-1-yl)-Urea

[0247] A solution of 3-(difluoromethoxy)benzylamine (12 mg, 0.07 mmol, 1.0 eq) in MeCN (0.4 mL) was treated at rt with DIPEA (43 .Math.L, 0.25 mmol, 3.5 eq) followed by CDI (12 mg, 0.07 mmol, 1.05 eq) and the resulting mixture was stirred at 50° C. for 30 min. The resulting mixture was treated with 3-fluorobicyclo[1.1.1]pentan-1-amine hydrochloride (10 mg, 0.07 mmol, 1.0 eq) and the reaction mixture was stirred at 80° C. overnight. The mixture was allowed to cool to rt and purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 0.95 min; [M+H].sup.+: 301.2.

[0248] Example 21 to Example 22 were synthesized using the appropriate amine or amine salt (HCI or TFA) derivative and following the procedure described in Example 20. LC-MS data of Example 21 to Example 22 are listed in the table below. The LC-MS conditions used were LC-MS (1).

TABLE-US-00009 Example N° Name t.sub.R [M+H].sup.+ 21 1-(3-Difluoromethoxy-benzyl)-3-(3-difluoromethyl-cyclobutyl)-urea 0.94 321.2 22 1-(3-Difluoromethoxy-benzyl)-3-(3-trifluoromethyl-bicyclo[1.1.1]pent-1-yl)-urea 1.08 351.2

Example 23: 1-(3-Trifluoromethoxy-Benzyl)-3-(3-Trifluoromethyl-Bicyclo[1.1.1]Pent-1-yl)-Urea

[0249] A solution of 3-(trifluoromethoxy)benzylamine (22 mg, 0.07 mmol, 1.0 eq) in MeCN (0.4 mL) was treated at rt with DIPEA (43 .Math.L, 0.25 mmol, 3.5 eq) followed by CDI (12 mg, 0.07 mmol, 1.05 eq) and the resulting mixture was stirred at 50° C. for 30 min. The resulting mixture was treated with 3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-amine hydrochloride (14 mg, 0.07 mmol, 1.0 eq) and the reaction mixture was stirred at 80° C. overnight. The mixture was allowed to cool to rt and purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 1.17 min; [M+H].sup.+: 369.2.

[0250] Example 24 to Example 26 were synthesized using the appropriate amine or amine salt (HCI or TFA) derivative and following the procedure described in Example 23. LC-MS data of Example 24 to Example 26 are listed in the table below. The LC-MS conditions used were LC-MS (1).

TABLE-US-00010 Example N° Name t.sub.R [M+H].sup.+ 24 1-(3-Fluoro-bicyclo[1.1.1]pent-1-yl)-3-(3-trifluoromethoxybenzyl)-urea 1.05 319.2 25 1-(3-Difluoromethyl-cyclobutyl)-3-(3-trifluoromethoxy-benzyl)-urea 1.03 339.2 26 1-Bicyclo[1.1.1]pent-1-yl-3-(3-trifluoromethoxy-benzyl)-urea 1.06 301.2

Example 27: 1-(3-Difluoromethyl-Benzyl)-3-(3-Difluoromethyl-Cyclobutyl)-Urea

[0251] To a solution of [3-(difluoromethyl)phenyl]methanamine hydrochloride (14 mg, 0.07 mmol, 1.0 eq) in MeCN (0.4 mL), DIPEA (43 .Math.L, 0.25 mmol, 3.5 eq) and CDI (17 mg, 0.11 mmol, 1.5 eq) were added in sequence. The resulting mixture was stirred at 50° C. for 1.5 h. 3-(difluoromethyl)cyclobutan-1-amine hydrochloride (11 mg, 0.07 mmol, 1.0 eq) was added. The reaction mixture was stirred at 80° C. overnight. The mixture was allowed to cool to rt and purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 0.92 min; [M+H].sup.+: 305.2.

Example 28: 1-(3-Difluoromethyl-Benzyl)-3-(3-Trifluoromethyl-Bicyclo[1.1.1]Pent-1-yl)-urea

[0252] The product was synthesized using 3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-amine hydrochloride and following the procedure described in Example 27. LC-MS (1): t.sub.R = 1.06 min; [M+H].sup.+: 335.2.

Example 29: 1-(3-Difluoromethyl-Cyclobutyl)-3-(2-Trifluoromethoxy-Pyridin-4-Ylmethyl)-Urea

[0253] To a solution of (2-(trifluoromethoxy)pyridin-4-yl)methanamine hydrochloride (23 mg, 0.07 mmol, 1.0 eq) in MeCN (0.4 mL), DIPEA (43 .Math.L, 0.25 mmol, 3.5 eq) and CDI (12 mg, 0.07 mmol, 1.05 eq) were added in sequence. The resulting mixture was stirred at 50° C. for 1.5 h. 3-(difluoromethyl)cyclobutan-1-amine hydrochloride (11 mg, 0.07 mmol, 1.0 eq) was added. The reaction mixture was stirred at 80° C. overnight. The mixture was allowed to cool to rt and purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 0.91 min; [M+H].sup.+: 340.2.

Example 30: 1-(2-Trifluoromethoxy-Pyridin-4-Ylmethyl)-3-(3-Trifluoromethyl-Bicyclo[1.1.1]Pent-1-yl)-Urea

[0254] The product was synthesized using 3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-amine hydrochloride and following the procedure described in Example 29. LC-MS (1): t.sub.R = 1.06 min; [M+H].sup.+: 370.2.

Example 31: (±)-1-(3-Difluoromethyl-Cyclobutyl)-3-{2-Methoxy-1-[2-(2,2,2-Trifluoro-Ethoxy)-Pyridin-4-yl]-Ethyl}-Urea

[0255] To a solution of (±)-2-methoxy-1-(2-(2,2,2-trifluoroethoxy)pyridin-4-yl)ethan-1-amine hydrochloride (30 mg, 0.11 mmol, 1.0 eq) in MeCN (0.6 mL), DIPEA (64 .Math.L, 0.37 mmol, 3.5 eq) and CDI (18 mg, 0.11 mmol, 1.05 eq) were added in sequence. The resulting mixture was stirred at 50° C. for 3 h. 3-(difluoromethyl)cyclobutan-1-amine hydrochloride (17 mg, 0.11 mmol, 1.0 eq) was added. The reaction mixture was stirred at 80° C. overnight. The mixture was allowed to cool to rt and purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 1.00 min; [M+H].sup.+: 398.3.

Example 32: (±)-1-{2-Methoxy-1-[2-(2,2,2-Trifluoro-Ethoxy)-Pyridin-4-yl]-Ethyl}-3-(3-Trifluoromethyl-Cyclobutyl)-Urea

[0256] The product was synthesized using 3-(trifluoromethyl)cyclobutan-1-amine hydrochloride and following the procedure described in Example 31. LC-MS (1): t.sub.R = 1.07 min; [M+H].sup.+: 416.3.

Example 33: (±)-1-[1-(2-Difluoromethoxy-Pyridin-4-yl)-Ethyl]-3-(3-Difluoromethyl-Cyclobutyl)-Urea

[0257] To a solution of (±)-1-(2-(difluoromethoxy)pyridin-4-yl)ethan-1-amine (22 mg, 0.07 mmol, 1.0 eq) in MeCN (0.4 mL), DIPEA (43 mL, 0.245 mmol, 3.5 eq) and CDI (12 mg, 0.07 mmol, 1.05 eq) were added in sequence. The resulting mixture was stirred at 50° C. for 3 h. 3-(difluoromethyl)cyclobutan-1-amine hydrochloride (11 mg, 0.07 mmol, 1.0 eq) was added. The reaction mixture was stirred at 80° C. overnight. The mixture was allowed to cool to rt and purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 0.93 min; [M+H].sup.+: 336.3.

Example 34: (±)-1-{1-[2-Methyl-6-(2,2,2-Trifluoro-Ethoxy)-Pyrimidin-4-yl]-Ethyl}-3-(3-Trifluoromethyl-Bicyclo[1.1.1]Pent-1-yl)-Urea

[0258] To a solution 3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-amine hydrochloride (126 mg, 0.64 mmol, 1.2 eq) in MeCN (2 mL), DIPEA (0.186 mL, 1.06 mmol, 2.0 eq) and CDI (103 mg, 0.638 mmol, 1.2 eq) were added in sequence. The mixture was stirred at 50° C. for 1 h. A solution of (±)-1-(2-methyl-6-(2,2,2-trifluoroethoxy)pyrimidin-4-yl)ethan-1-amine (125 mg, 0.531 mmol, 1.0 eq) in MeCN (1.1 mL) was added. The reaction mixture was stirred at 80° C. overnight. The mixture was allowed to cool to rt and purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 1.12 min; [M+H].sup.+: 413.3.

Example 35: 1-Bicyclo[1.1.1]Pent-1-yl-3-(3-Trifluoromethyl-Benzyl)-Urea

[0259] To a solution of bicyclo[1.1.1]pentan-1-amine hydrochloride (10 mg, 0.08 mmol, 1 eq) in THF (1 mL), NEt.sub.3 (45 .Math.L, 0.32 mmol, 4 eq) and 4-nitrophenyl (3-(trifluoromethyl)benzyl)carbamate (27 mg, 0.08 mmol, 1 eq) were added in sequence. The resulting mixture was stirred at rt overnight. The mixture was concentrated in vacuo. The residue was purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 1.03 min; [M+H].sup.+: 285.2.

[0260] Example 36 to Example 42 were synthesized using the appropriate amine or amine salt (HCI or TFA) derivative and following the procedure described in Example 35. LC-MS data of Example 36 to Example 42 are listed in the table below. The LC-MS conditions used were LC-MS (1).

TABLE-US-00011 Example N° Name t.sub.R [M+H].sup.+ 36 1-(3-Fluoro-bicyclo[1.1.1]pent-1-yl)-3-(3-trifluoromethyl-benzyl)-urea 1.02 303.2 37 1-(3-Trifluoromethyl-benzyl)-3-(3-trifluoromethyl-bicyclo[1.1.1]pent-1-yl)-urea 1.14 353.2 38 1-(3-Difluoromethyl-cyclobutyl)-3-(3-trifluoromethyl-benzyl)-urea 1.00 323.2 39 1-(3-Methyl-bicyclo[1.1.1]pent-1-yl)-3-(3-trifluoromethyl-benzyl)-urea 1.10 299.2 40 1-(3-Fluoromethyl-bicyclo[1.1.1]pent-1-yl)-3-(3-trifluoromethyl-benzyl)-urea 1.02 317.2 41 1-(3-Trifluoromethyl-benzyl)-3-(3-trifluoromethyl-cyclobutyl)-urea 1.07 341.2 42 1-(3-Hydroxy-3-trifluoromethyl-cyclobutyl)-3-(3-trifluoromethyl-benzyl)-urea 0.94 357.2

Example 43: 1-Bicyclo[1.1.1]Pent-1-yl-3-[2-(2,2,2-Trifluoro-Ethoxy)-Pyridin-4-Ylmethyl]-Urea

[0261] To a solution of bicyclo[1.1.1]pentan-1-amine hydrochloride (30 mg, 0.08 mmol, 1 eq) in THF (1 mL), NEt.sub.s (45 .Math.L, 0.32 mmol, 4 eq) and 4-nitrophenyl ((2-(2,2,2-trifluoroethoxy)pyridin-4-yl)methyl)carbamate (30 mg, 0.08 mmol, 1 eq) were added in sequence. The resulting mixture was stirred at rt overnight. The mixture was concentrated in vacuo. The residue was purified by prep. HPLC (column: Waters XBridge, 30×75 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 0.98 min; [M+H].sup.+: 316.2.

[0262] Example 44 to Example 46 were synthesized using the appropriate amine or amine salt (HCI or TFA) derivative and following the procedure described in Example 43. LC-MS data of Example 44 to Example 46 are listed in the table below. The LC-MS conditions used were LC-MS (1).

TABLE-US-00012 Example N° Name t.sub.R [M+H].sup.+ 44 1-(3-Fluoro-bicyclo[1.1.1]pent-1-yl)-3-[2-(2,2,2-trifluoro-ethoxy)-pyridin-4-ylmethyl]-urea 0.97 334.2 45 1-[2-(2,2,2-Trifluoro-ethoxy)-pyridin-4-ylmethyl]-3-(3-trifluoromethyl-bicyclo[1.1.1]pent-1-yl)-urea 1.10 384.2 46 1-(3-Difluoromethyl-cyclobutyl)-3-[2-(2,2,2-trifluoro-ethoxy)-pyridin-4-ylmethyl]-urea 0.96 354.2

Example 47: 1-(3-Difluoromethyl-Bicyclo[1.1.1]Pent-1-yl)-3-(3-Trifluoromethyl-Benzyl)-Urea

[0263] To an ice-cooled solution of 3-(difluoromethyl)bicyclo[1.1.1]pentan-1-amine hydrochloride (125 mg, 0.74 mmol, 1 eq) in DCM (20 mL), NEt.sub.s (0.31 mL, 2.21 mmol, 3 eq) and 1-(isocyanatomethyl)-3-(trifluoromethyl)benzene (156 mg, 0.74 mmol, 1 eq) were added dropwise in sequence. The resulting mixture was stirred at 0° C. for 2 hours. The mixture was diluted with sat. aq. NaHCO.sub.3 soln. and extracted with DCM (3x). The comb. org. layers were washed with sat. aq. NaCl soln., dried over MgSO.sub.4, filtered and concentrated in vacuo. The residue was purified by prep. HPLC (column : Waters XBridge, 30×50 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 1.06 min; [M+H].sup.+: 335.2.

Example 48: 1-Bicyclo[2.1.1]Hex-1-yl-3-(3-Trifluoromethyl-Benzyl)-Urea

[0264] To a solution of bicyclo[2.1.1]hexan-1-amine hydrochloride (27 mg, 0.2 mmol, 1.0 eq) in MeCN (0.7 mL), DIPEA (87 .Math.L, 0.5 mmol, 2.5 eq) and 1-(isocyanatomethyl)-3-(trifluoromethyl)benzene (64 mg, 0.3 mmol, 1.5 eq) were added in sequence. The reaction mixture was stirred at rt overnight. The mixture was purified by prep. HPLC (column : Waters XBridge, 30×50 mm, 10 um, UV/MS, basic conditions). LC-MS (1): t.sub.R = 1.09 min; [M+H].sup.+: 299.2.

Example 49: 1-(3,3-Difluoro-1-Methyl-Cyclobutyl)-3-(3-Trifluoromethyl-Benzyl)-Urea

[0265] The product was synthesized using 3,3-difluoro-1-methylcyclobutanamine hydrochloride and following the procedure described in Example 48. LC-MS (1): t.sub.R = 1.06 min; [M+H].sup.+: 323.2.

Example 50: 1-(3-(Trifluoromethyl)Benzyl)-3-(3-(Trifluoromethyl)Cyclobutyl)Urea (Stereoisomer 1) and Example 51: 1-(3-(Trifluoromethyl)Benzyl)-3-(3-(Trifluoromethyl)Cyclobutyl)Urea (Stereoisomer 2)

[0266] 1-Trifluoromethyl-benzyl)-3-trifluoromethyl-cyclobutyl)-urea was separated by the preparative chiral SFC 1 method to give Example 50 (first eluting, t.sub.R = 2.9 min) and Example 51 (second eluting, t.sub.R = 4.0 min). Example 50 (LC-MS (1): t.sub.R = 1.07 min; [M+H].sup.+: 341.2), .sup.1H-NMR (500 MHz, DMSO) δ: 7.54-7.59 (m, 4 H), 6.51-6.53 (m, 2 H), 4.27 (d, J = 6.1 Hz, 2 H), 4.17-4.23 (m, 1 H), 2.85-3.12 (m, 1 H), 2.33-2.38 (m, 2 H), 2.18-2.24 (m, 2 H). Example 51 (LC-MS (1): t.sub.R = 1.07 min; [M+H].sup.+: 341.2), .sup.1H-NMR (500 MHz, DMSO) δ: 7.53-7.59 (m, 4 H), 6.48-6.65 (m, 1 H), 6.39-6.48 (m, 1 H), 4.28 (d, J = 6.1 Hz, 2 H), 3.92-4.19 (m, 1 H), 2.76-2.88 (m, 1 H), 2.35-2.42 (m, 2 H), 1.88-1.94 (m, 2 H).

Example 52: 1-(3-(Difluoromethyl)Cyclobutyl)-3-(3-(Trifluoromethyl)Benzyl)Urea (Stereoisomer 1) and Example 53: 1-(3-(Difluoromethyl)Cyclobutyl)-3-(3-(Trifluoromethyl)Benzyl)Urea (Stereoisomer 2)

[0267] 1-Difluoromethyl-cyclobutyl)-3-trifluoromethyl-benzyl)-urea was separated by the preparative chiral SFC 2 method to give Example 52 (first eluting, t.sub.R = 2.1 min) and Example 53 (second eluting, t.sub.R = 2.6 min). Example 52 (LC-MS (1): t.sub.R = 1.01 min; [M+H].sup.+: 323.2) );.sup.1 H-NMR (500 mHz, DMSO) δ: 7.54-7.59 (m, 4 H), 6.38-6.43 (m, 2 H), 5.99 (td, J = 57.3, 4.2 Hz, 1 H), 4.28 (d, J = 6.1 Hz, 2 H), 4.05 (h, J = 8.4 Hz, 1 H), 2.31-2.43 (m, 1 H), 2.23-2.28 (m, 2 H), 1.76-1.83 (m, 2 H). Example 53 (LC-MS (1): t.sub.R = 1.01 min; [M+H].sup.+: 323.2); .sup.1H-NMR (500 MHz, DMSO) δ: 7.54-7.59 (m, 4 H), 6.44-6.47 (m, 2 H), 6.18 (td, J = 57.2, 4.8 Hz, 1 H), 4.27 (d, J = 6.1 Hz, 2 H), 4.18 (h, J = 7.9 Hz, 1 H), 2.47-2.57 (m, 1 H), 2.21-2.26 (m, 2 H), 2.01-2.07 (m, 2 H).

Example 54: 1-{(S)-1-[2-Methyl-6-(2,2,2-Trifluoro-Ethoxy)-Pyrimidin-4-yl]-Ethyl}-3-(3-Trifluoromethyl-Bicyclo[1.1.1]Pent-1-yl)-Urea

[0268] Racemic 1-{1-[2-Methyl-6-(2,2,2-trifluoro-ethoxy)-pyrimidin-4-yl]-ethyl}-3-(3-trifluoromethyl-bicyclo[1.1.1]pent-1-yl)-urea was separated by the preparative chiral SFC 3 method to give Example 54 (first eluting, t.sub.R = 1.6 min) and 1-{(R)-1-[2-Methyl-6-(2,2,2-trifluoro-ethoxy)-pyrimidin-4-yl]-ethyl}-3-(3-trifluoromethyl-bicyclo[1.1.1]pent-1-yl)-urea (second eluting, t.sub.R = 2.2 min). Example 54 (LC-MS (1): t.sub.R = 1.12 min; [M+H].sup.+: 413.3). The stereochemistry at the benzylic position has been assigned in analogy to Example 190 of PCT/EP2021/060918, meaning that the more active isomer was assumed to have (S)-configuration.

Example 55: 1-(3-(Difluoromethyl)Cyclobutyl)-3-((2-(2,2,2-Trifluoroethoxy)Pyridin-4-yl)Methyl)Urea (Stereoisomer 1) and Example 56: 1-(3-(Difluoromethyl)Cyclobutyl)-3-((2-(2,2,2-Trifluoroethoxy)Pyridin-4-yl)Methyl)Urea (Stereoisomer 2)

[0269] 1-Difluoromethyl-cyclobutyl)-3-[2-(2,2,2-trifluoro-ethoxy)-pyridin-4-ylmethyl]-urea was separated by the preparative chiral SFC 4 method to give Example 55 (first eluting, t.sub.R = 3.6 min) and Example 56 (second eluting, t.sub.R = 4.6 min). Example 55 (LC-MS (1): t.sub.R = 0.96 min; [M+H].sup.+: 354.3), .sup.1H-NMR (500 MHz, DMSO) δ: 8.10-8.11 (m, 1 H), 6.96 (dd, J = 5.3, 1.2 Hz, 1 H), 6.76 (br s, 1 H), 6.39-6.45 (m, 2 H), 6.00 (td, J = 57.3, 4.2 Hz, 1 H), 4.98 (q, J = 9.1 Hz, 2 H), 4.20 (d, J = 6.1 Hz, 2 H), 4.05 (h, J = 8.5 Hz, 1 H), 2.32-2.45 (m, 1 H), 2.17-2.32 (m, 2 H), 1.77-1.83 (m, 2 H). Example 56 (LC-MS (1): t.sub.R = 0.96 min; [M+H].sup.+: 354.2), .sup.1H-NMR (500 MHz, DMSO) δ: 8.10-8.11 (m, 1 H), 6.97 (dd, J = 5.2, 1.2 Hz, 1 H), 6.76 (br s, 1 H), 6.43-6.51 (m, 2 H), 6.18 (td, J = 57.2, 4.8 Hz, 1 H), 4.98 (q, J = 9.1 Hz, 2 H), 4.15-4.21 (m, 3 H), 2.53-2.59 (m, 1 H), 2.22-2.27 (m, 2 H), 1.92-2.13 (m, 2 H).

Example 57: 1-(3-(Difluoromethyl)Cyclobutyl)-3-((S)-1-(3-(Trifluoromethoxy)Phenyl)Ethyl)Urea (Stereoisomer 1) and Example 58: 1-(3-(Difluoromethyl)Cyclobutyl)-3-((S)-1-(3-(Trifluoromethoxy)Phenyl)Ethyl)Urea (Stereoisomer 2)

[0270] 1-Difluoromethyl-cyclobutyl)-3-[1-trifluoromethoxy-phenyl)-ethyl]-urea was first separated by preparative chiral SFC 4 method to give Fraction 1 (first eluting, t.sub.R = 1.8 min) and Fraction 2 (second eluting, t.sub.R = 2.9 min). Fraction 1 was further separated by preparative chiral SFC 1 method to give Example 57 (first eluting, t.sub.R = 2.8 min) and Example 58 (second eluting, t.sub.R = 3.8 min). Example 57 (LC-MS (1): t.sub.R = 1.08 min; [M+H].sup.+: 353.2), .sup.1H-NMR (500 MHz, DMSO) δ: 7.45 (t, J = 7.8 Hz, 1 H), 7.31 (d, J = 7.8 Hz, 1 H), 7.19-7.24 (m, 2 H), 6.35 (d, J = 8.0 Hz, 1 H), 6.16 (d, J = 8.3 Hz, 1 H), 5.99 (td, J = 57.3, 4.2 Hz, 1 H), 4.76 (quint, J = 7.1 Hz, 1 H), 3.98-4.03 (m, 1 H), 2.32-2.42 (m, 1 H), 2.20-2.26 (m, 2 H), 1.72-1.79 (m, 2 H), 1.31 (d, J = 7.1 Hz, 3 H). Example 58 (LC-MS (1): t.sub.R = 1.08 min; [M+H].sup.+: 353.3), .sup.1H-NMR (500 MHz, DMSO) δ: 7.45 (t, J = 7.9 Hz, 1 H), 7.32 (d, J = 7.9 Hz, 1 H), 7.19-7.24 (m, 2 H), 6.39 (d, J = 8.0 Hz, 1 H), 6.23 (d, J = 8.0 Hz, 1 H), 6.17 (td, J = 57.2, 4.8 Hz, 1 H), 4.74-4.79 (m, 1 H), 4.10-4.16 (m, 1 H), 2.48-2.53 (m, 1 H), 2.17-2.24 (m, 2 H), 1.97-2.04 (m, 2 H), 1.31 (d, J = 7.0 Hz, 3 H). Fraction 2 was further separated by preparative chiral SFC 6 method to give 1-(3-(difluoromethyl)cyclobutyl)-3-((R)-1-(3-(trifluoromethoxy)phenyl)ethyl)urea (first eluting stereoisomer, t.sub.R = 1.7 min) and 1-(3-(difluoromethyl)cyclobutyl)-3-((R)-1-(3-(trifluoromethoxy)phenyl)ethyl)urea (second eluting stereoisomer, t.sub.R = 2.3 min). The stereochemistry at the benzylic position has been assigned in analogy to Example 190 of PCT/EP2021/060918, meaning that the more active isomer was assumed to have (S)-configuration.

Synthesis of 4-nitrophenyl (3-(trifluoromethyl)benzyl)carbamate

[0271] To an ice-cooled solution of 3-(trifluoromethyl)benzylamine (1.50 g, 8.4 mmol, 1 eq) and DIPEA (4.31 mL, 25.2 mmol, 3 eq) in THF (43 mL), 4-nitrophenyl chloroformate (1.74 g, 8.4 mmol, 1 eq) was added. The resulting mixture was stirred at 0° C. for 1 hour. The reaction mixture was diluted with water (25 mL) and EtOAc (25 mL). The layers were separated. The aq. phase was extracted with EtOAc (2 × 25 mL). The comb. org. phases were dried over MgSO.sub.4 and concentrated in vacuo. The residue was purified by CombiFlash (column: 40 g, flow: 37 mL/min, Heptane 100% to Heptane + 20% EtOAc) to afford a pale yellow solid which was further triturated in heptane/EtOAc 8:2 to yield a white solid. LC-MS (2): t.sub.R = 1.00 min; no ionization.

[0272] The following carbamate was synthesized using the appropriate amine and following the procedure described for 4-nitrophenyl (3-(trifluoromethyl)benzyl)carbamate. LC-MS data are listed in the table below. The LC-MS conditions used were LC-MS (2).

TABLE-US-00013 Name t.sub.R [M+H].sup.+ 4-nitrophenyl ((2-(2,2,2-trifluoroethoxy)pyridin-4-yl)methyl)carbamate 0.97 372.15

Synthesis of (3-(2,2,2-Trifluoroethoxy)Phenyl)Methanamine

Step 1: 3-(2,2,2-Trifluoroethoxy)Benzaldehyde

[0273] To a solution of 3-hydroxybenzaldehyde (3.0 g, 24.6 mmol, 1.0 eq) and Cs.sub.2CO.sub.3 (12.0 g, 36.8 mmol, 1.5 eq) in DMF (45 mL), trifluoromethansulfonic acid 2.2,2-trifluoroethylester (4.25 ml, 29.5 mmol, 1.2 eq) was added dropwise. The reaction was stirred at rt for 2 h. The reaction was quenched with water and the mixture was extracted with Et;O. The comb. org. phases were dried over MgSO.sub.4 and concentrated in vacuo to yield an orange oil. The product was used without further purification. LC-MS (3): t.sub.R = 0.75 min; no ionization.

Step 2: (:I:, E)-2-Methyl-N-(3-(2, 2, 2-Trifluoroethoxy)Benzylidene) Propane-2-Sulfinamide

[0274] A mixture of 3-(2,2,2-trifluoroethoxy)benzaldehyde (5.01 g, 24.5 mmol, 1 eq), (±)-2-methyl-2-propanesulfinamide (2.98 g, 24.5 mmol, 1 eq), and Ti(OEt).sub.4 (10.3 mL, 49.1 mmol, 2 eq) in THF (42 mL) was stirred at rt for 3 d. The reaction was quenched with sat. aq. NaCl soln. The resulting suspension was filtered and the solids rinsed with EtOAc. The filtrate was washed with sat. aq. NaCl soln., dried over MgSO.sub.4, and concentrated in vacuo. The residue was purified by flash column chromatography (SiO.sub.2, DCM) to give an orange solid. LC-MS (3): t.sub.R = 0.87 min; [M+H].sup.+: 307.98.

Step 3: (±)-2-Methyl-N-(3-(2,2,2-Trifluoroethoxy)Benzyl)Propane-2-Sulfinamide

[0275] To a solution of (±,E)-2-methyl-N-(3-(2,2,2-trifluoroethoxy)benzylidene)propane-2-sulfinamide (6.85 g, 22.3 mmol, 1 eq) in MeOH (78 mL) and DCM (162 mL), NaBH.sub.4 (5.06 g, 134 mmol, 6 eq) was added. The reaction mixture was stirred at rt for 10 min. The reaction was quenched with water. The mixture was extracted with DCM. The comb. org. phases were dried over MgSO.sub.4 and concentrated in vacuo to afford a colorless oil. The product was used without further purification. LC-MS (3): t.sub.R = 0.75 min; [M+H].sup.+: 310.00.

Step 4: (3-(2,2,2-Trifluoroethoxy)Phenyl)Methanamine

[0276] To an ice-cooled solution of (±)-2-methyl-N-(3-(2,2,2-trifluoroethoxy)benzyl)propane-2-sulfinamide (6.65 g, 21.5 mmol, 1 eq) in anhydrous methanol (80 mL), 4N HCI in dioxane (10.8 mL, 43 mmol, 2 eq) was added dropwise. The reaction mixture was stirred at 0° C. for 10 min and further at rt overnight. The yellow homogeneous reaction mixture was carefully concentrated to dryness under reduced pressure. The residue was partitioned between DCM (150 mL) and water (30 mL). Solid Na.sub.2CO.sub.3 (11.39 g, 107 mmol, 5 eq) was added. The layers were separated and the aq. phase was extracted with DCM (50 mL). The comb. org. phases were dried over MgSO.sub.4 and concentrated in vacuo. The product was used without further purification. LC-MS (3): t.sub.R = 0.46 min; [M+H].sup.+: 206.06.

Synthesis of 1-(3-(Difluoromethoxy)Phenyl)Ethan-1-Amine Hydrochloride

[0277] To a solution of 3-(difluoromethoxy)benzonitrile (1.0 g, 5.79 mmol, 1 eq) in THF (5 mL), 3.4 M methylmagnesium bromide in 2-methyltetrahydrofuran (5.11 mL, 17.4 mmol, 3 eq) was added dropwise. The mixture was stirred at rt for 2 hours. The reaction was cooled to 15° C. and quenched with MeOH (20 mL). NaBH.sub.4 (438 mg, 11.6 mmol, 2 eq) was added and the mixture was stirred at rt overnight. 2 M aq. HCI soln. (30 mL) was added and the mixture was stirred at rt for 5 min. The organic solvents were removed in vacuo. The resulting solution was partitioned between DCM (50 mL) and sat. aq. NaHCO.sub.3 soln. (30 mL). The layers were separated. The org. phase was treated with 1.25 M HCI in MeOH (20 mL) and concentrated in vacuo to give an oil. The product was used without further purification. LC-MS (4): t.sub.R = 0.73 min; [M+H].sup.+: 188.34.

Synthesis of (2-(Trifluoromethoxy)Pyridin-4-yl)Methanamine Hydrochloride

Step 1: (±,E)-2-Methyl-N-((2-(Trifluoromethoxy)Pyridin-4-yl)Methylene)Propane-2-Sulfinamide

[0278] To a mixture of 2-(trifluoromethoxy)pyridine-4-carbaldehyde (573 mg, 3 mmol, 1.0 eq) and (±)-2-methylpropane-2-sulfinamide (498 mg, 3.9 mmol, 1.3 eq) in THF (15 mL), titanium ethoxide (0.68 mL, 3.3 mmol, 1.1 eq) was added dropwise. The solution was stirred at rt for 17 hours. The yellow solution was diluted with water (20 mL) and DCM (10 mL). The resulting mixture was filtered. The layers were separated and the aq. phase was extracted with DCM (2 × 20 mL). The comb. org. phases were washed with H.sub.2O (1 × 20 mL), sat. aq. NaCl soln. (1 × 20 mL), dried over MgSO.sub.4, and concentrated in vacuo. The residue was purified by Combiflash (column: 40 g, flow: 40 mL/min, heptane to heptane/EtOAc 100:30) to give a white solid. LC-MS (2): t.sub.R = 0.96 min; [M+H].sup.+: 295.18.

Step 2: (±)-2-Methyl-N-((2-(Trifluoromethoxy)Pyridin-4-yl)Methyl)Propane-2-Sulfinamide

[0279] To an ice-cooled solution of (±,E)-2-methyl-N-((2-(trifluoromethoxy)pyridin-4-yl)methylene)propane-2-sulfinamide (285 mg, 0.97 mmol, 1.0 eq) in MeOH (20 mL), sodium borohydride (55 mg, 1.45 mmol, 1.5 eq) was added. The mixture was stirred at 0° C. for 2.5 hours. The reaction mixture was concentrated in vacuo. The residue was partitioned between water (25 mL) and DCM (25 mL). The layers were separated. The aq. phase was extracted with DCM (2 × 25 mL). The comb. org. phases were dried over MgSO.sub.4 and concentrated in vacuo. The product was used without further purification. LC-MS (2): t.sub.R = 0.82 min; [M+H].sup.+: 297.22.

Step 3: (2-(Trifluoromethoxy)Pyridin-4-yl)Methanamine Hydrochloride

[0280] To an ice-cooled solution of (±)-2-methyl-N-((2-(trifluoromethoxy)pyridin-4-yl)methyl)propane-2-sulfinamide (279 mg, 0.94 mmol, 1 eq) in MeOH (20 mL), 4N HCI in dioxane (1.2 mL, 4.71 mmol, 5 eq) was added. The resulting mixture was stirred at 0° C. for 1 hour. The reaction mixture was concentrated in vacuo. The product was used crude for the next step. LC-MS (2): t.sub.R = 0.40 min; [M+H].sup.+: 193.28.

Synthesis of (±)-2-Methoxy-1-(2-(2,2,2-Trifluoroethoxy)Pyridin-4-yl)Ethan-1-Amine hydrochloride

Step 1: Tert-Butyl (±)-(2-Methoxy-1-(2-(2,2,2-Trifluoroethoxy)Pyridin-4-yl)Ethyl)Carbamate

[0281] To a solution of 4-bromo-2-(2,2,2-trifluoroethoxy)pyridine (571 mg, 2.17 mmol, 1.0 eq) in degassed DMSO (100 mL), 2-{[(tert-butoxy)carbonyl]amino}-3-methoxypropanoic acid (749 mg, 3.25 mmol, 1.5 eq), potassium phosphate tribasic (1.41 g, 6.5 mmol, 3.0 eq), 4,4′-di-tert-butyl-2,2′-dipyridyl (59 mg, 0.217 mmol, 0.1 eq), NiCl.sub.2.Math.glyme (49 mg, 0.217 mmol, 0.1 eq) and [lr{dF(CF.sub.3)ppy}.sub.2(dtbpy)]PF.sub.s (49 mg, 0.04 mmol, 0.02 eq) were added in sequence. The resulting mixture was degassed with N.sub.2 while stirring for 15 minutes. Then, the resulting mixture was stirred at rt overnight under blue LED irradiation. Water was added and the mixture was extracted with EtOAc (3x). The comb. org. layers were further washed with sat. aq. NaCl soln., dried over MgSO.sub.4, filtered and concentrated in vacuo. The residue was purified by Combiflash (column: 24 g, flow: 25 mL/min, Heptane to Heptane + 30% EtOAc) to give an yellow oil. LC-MS (2): t.sub.R = 0.98 min; [M+H].sup.+: 351.25.

Step 2: (±)-2-Methoxy-1-(2-(2,2,2-Trifluoroethoxy)Pyridin-4-yl)Ethan-1-Amine Hydrochloride

[0282] A solution of tert-butyl (±)-(2-methoxy-1-(2-(2,2,2-trifluoroethoxy)pyridin-4-yl)ethyl)carbamate (600 mg, 1.71 mmol, 1 eq) in 4N HCI in dioxane (6.85 mL, 27.4 mmol, 16 eq) was stirred at rt overnight. The mixture was concentrated in vacuo to afford a white solid. The product was used without further purification. LC-MS (2): t.sub.R = 0.56 min; [M+H].sup.+: 251.25.

Synthesis of (±)-1-(2-(Difluoromethoxy)Pyridin-4-yl)Ethan-1-Amine

Step 1: 1-(2-(Difluoromethoxy)Pyridin-4-yl)Ethan-1-One

[0283] To an ice-cooled solution of 2-(difluoromethoxy)pyridine-4-carbonitrile (1.50 g, 8.38 mmol, 1.0 eq) in THF (80 mL), 3 M methylmagnesium bromide solution (6.13 mL, 18.4 mmol, 2.2 eq) was added dropwise. The resulting mixture was stirred at rt overnight. The resulting mixture was quenched with 1 M HCI aq. soln. (15 mL) and the resulting mixture was stirred at rt for 1 h. The reaction mixture was diluted with sat. aq. NaHCO.sub.3 soln. and EtOAc. The layers were separated and the aq. phase was extracted with EtOAc (1× 30 mL). The comb. org. phases were washed with sat. aq. NaCl soln. (1 × 20 mL), dried over MgSO.sub.4, and concentrated in vacuo. The residue was purified by Combiflash (column: 40 g, flow: 40 mL/min, Heptane to Heptane + 18% EtOAc) to give a colorless oil. LC-MS (2): t.sub.R = 0.78 min; [M+H].sup.+: 188.26.

Step 2: (±)-1-(2-(Difluoromethoxy)Pyridin-4-yl)Ethan-1-Amine

[0284] To a solution of 1-(2-(difluoromethoxy)pyridin-4-yl)ethan-1-one (610 mg, 3.26 mmol, 1 eq) in MeOH (100 mL), ammonium acetate (5.03 g, 65.2 mmol, 20 eq) and sodium cyanoborohydride (410 mg, 6.52 mmol, 2 eq) were added in sequence. The resulting mixture was stirred at rt overnight. The resulting mixture was concentrated in vacuo.The residue was diluted with sat. aq. NaHCO.sub.3 soln. and DCM. The layers were separated and the aq. phase was extracted with DCM (1× 30 mL). The comb. org. phases were washed with sat. aq. NaCl soln. (1 × 20 mL), dried over MgSO.sub.4, and concentrated in vacuo. The residue was used without further purification. LC-MS (2): t.sub.R = 0.43 min; [M+H].sup.+: 189.31.

Synthesis of (±)-1-(2-Methyl-6-(2,2,2-Trifluoroethoxy)Pyrimidin-4-yl)Ethan-1-Amine

Step 1: 2-Methyl-6-(2,2,2-Trifluoroethoxy)Pyrimidine-4-Carbonitrile

[0285] To an ice-cooled solution of 2.2,2-trifluoroethanol (1.0 mL, 13.6 mmol, 2.2 eq) in THF (8 mL), sodium hydride 60 % dispersion in mineral oil (569 mg, 14.2 mmol, 2.3 eq) was added portionwise. The resulting mixture was warmed to rt over 30 min and stirred at rt for 30 min. The mixture was cooled to 0° C. and a solution of 6-chloro-2-methylpyrimidine-4-carbonitrile (1.0 g, 6.19 mmol, 1 eq) in THF (4 mL) was added dropwise. The resulting mixture was slowly warmed to rt and stirred at rt for 30 min. The reaction mixture was slowly poured into cold water (40 mL), and then extracted with EtOAc (2× 40 mL). The comb. org. layers were washed with sat. aq. NaCl soln., dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue was purified by Combiflash (column: 24 g, flow: 35 mL/min, Heptane to Heptane + 15% TBME) to give a pale yellow oil. LC-MS (2): t.sub.R = 0.86 min; [M+H].sup.+: 218.30.

Step 2: 1-(2-Methyl-6-(2,2,2-Trifluoroethoxy)Pyrimidin-4-yl)Ethan-1-One

[0286] To a solution cooled at -78° C. of 2-methyl-6-(2,2,2-trifluoroethoxy)pyrimidine-4-carbonitrile (945 mg, 4.35 mmol, 1.0 eq) in THF (37 mL), 3 M methylmagnesium bromide solution in diethyl ether (9.3 mL, 27.8 mmol, 6.4 eq) was added dropwise. The resulting mixture was stirred at rt for 1 hour. The resulting mixture was cooled to 0° C. and slowly quenched with 10% acetic acid aq. soln. (15 mL). The reaction mixture was diluted with sat. aq. NaHCO.sub.3 soln. (50 mL) and EtOAc (50 mL). The layers were separated and the aq. phase was extracted with EtOAc (1× 50 mL). The comb. org. phases were washed with sat. aq. NaCl soln. (1 × 50 mL), dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was purified by Combiflash (column: 40 g, flow: 40 mL/min, Heptane to Heptane + 12% TBME) to give a yellow solid. LC-MS (2): t.sub.R = 0.88 min; [M+H].sup.+: 235.28.

Step 3: (±)-1-(2-Methyl-6-(2,2,2-Trifluoroethoxy)Pyrimidin-4-yl)Ethan-1-Amine

[0287] To a solution of 1-(2-methyl-6-(2,2,2-trifluoroethoxy)pyrimidin-4-yl)ethan-1-one (271 mg, 1.08 mmol, 1 eq) in MeOH (31 mL), ammonium acetate (1.66 g, 21.5 mmol, 20 eq) and sodium cyanoborohydride (142 mg, 2.15 mmol, 2 eq) were added in sequence. The resulting mixture was stirred at rt overnight. The resulting mixture was concentrated in vacuo. The residue was diluted with sat. aq. NaHCO.sub.3 soln. (40 mL) and DCM (40 mL). The layers were separated and the aq. phase was extracted with DCM (1× 40 mL). The comb. org. phases were washed with sat. aq. NaCl soln. (1 × 40 mL), dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was purified by prep. HPLC (column : Waters XBridge, 30×50 mm, 10 um, UV/MS, basic conditions). LC-MS (4): t.sub.R = 0.74 min; [M+H].sup.+: 236.06.

II. Biological Assays

A) Rat Oscillation Assay Assay Principle

[0288] This assay is a functional phenotypic assay designed to mimic epileptic seizures using primary neuronal cultures from embryonic rat brains, which form a functional neuronal network that generate synchronized intracellular calcium concentration oscillations when cultured at high density in 384-well plate. The epileptic phenotype is induced by incubating the neurons in magnesium-free assay buffer, that results in increased probability of NMDA receptor activation, leading to an increased frequency and amplitude of intracellular calcium oscillations. Once neurons are incubated with the calcium indicator dye Fluo-8 AM (Tebu-bio), neuronal calcium oscillations can be monitored in real time using FLIPR® Tetra (fluorometric plate reader, Molecular Devices). With these recordings, the effect of anti-epileptic drugs can be quantified. The anti-epileptic effect of compounds, which activate directly or indirectly the Kv7 channels can be modulated by the Kv7 channel blocker XE-991. The assay was performed as described previously (Pacico N, Mingorance-Le Meur A. New In Vitro Phenotypic Assay for Epilepsy: Fluorescent Measurement of Synchronized Neuronal Calcium Oscillations. PLoS ONE 9(1) 2014) with modifications described hereafter.

Neuronal Cultures

[0289] Animal care followed standard procedures in accordance with swiss institutional guidelines. Dissociated neuronal cultures were obtained from cerebral cortices of embryonic Wistar rats at embryonic stage E18 (Charles River). The uterine horns were removed by caesarian surgery from deeply anesthetized rats (Isofurane) and sacrificed by decapitation. The embryos were decapitated by closing forceps. The brains were isolated and dissected one by one in ice-cold PBS (Life Technologies) under optical control using a binocular. Meninges, olfactory bulbs, and basal ganglia were removed. Cortical hemispheres (still including the hippocampus) were cut in small pieces with tweezers and placed on ice in prechilled Hibernate-E medium (Life technology). The hemispheres were then incubated in 10 mL of Hibernate-E containing 15 U/mL papain (Worthington) for 25 min at 30° C. with gentle mixing every 10 min. Genomic DNA was then digested by prolonging the incubation during 10 min at 37° C. in presence of 4 U/mL rDNase I (Ambion). The obtained suspension was then centrifuged at 800 g for 5 min and the cell pellet was resuspended in 2 mL Hibernate-E and gently dissociated by pipetting up and down 10 times with a plastic Pasteur pipette resulting in a homogenous cell suspension. This suspension was immediately filtered through a 70 .Math.m cell strainer (MACS® SmartStrainer, Miltenyi), collected in 10 mL Hibernate-E and centrifuged at 800 g for 5 min. The cell pellet was resuspended in Neurobasal medium, supplemented with 2% B-27, 0.5 mM Glutamax-I, 100 U/mL penicillin, 100 .Math.g/mL streptomycin (Life Technologies) and diluted at the final concentration of 300′000 cells/mL. One day before plating the cells, 384-well plates were coated with 25 .Math.L/well of 0.1 % poly-L-lysine solution (Sigma), incubated overnight at 37° C., washed two times with sterile distilled water and allowed to dry at room temperature for >2 h. The neurons were seeded at a density of 15′000 cells/well in 50 .Math.L/well in a 384-well black, clear-bottomed plates (Corning) and subsequently maintained in an incubator at 37° C., 5% CO.sub.2 and 95% humidity for 8 to 10 days. After 3 and 7 days, 40% of media was renewed under sterile conditions.

Protocol Rat Oscillation Assay

[0290] Neurons seeded in the assay plates were washed with Hank’s balanced salt solution (HBSS) devoid of Ca.sup.2+ and Mg.sup.2+, supplemented with 20 mM HEPES (Life Technologies) and 2 mM CaCl.sub.2 (Sigma), pH 7.4 (hereafter called Assay buffer) using a Biotek EL406 plate washer. Neurons were loaded with 1 .Math.M Fluo-8 AM in Assay buffer for 15 min at 37° C., 5% CO.sub.2. Buffer containing dye was then removed and the assay plates were washed 3 times with Assay buffer using the Biotek EL406 washer and allowed to equilibrate in 50 .Math.L of assay buffer at room temperature for 25 min. The kinetic curves of fluorescence fluctuations acquired once per second using FLIPR® Tetra reflect neuronal calcium oscillations. Recording was performed in two phases separated by 20 min resulting in two acquisitions: “Acute” and “20 min”. In the “Acute” acquisition phase, fluorescence was recorded over a period of 500 sec in presence or absence of the Kv7 channel blocker XE-991. Test compounds were added 250 sec after acquisition start. 20 min after compound addition, calcium oscillations were recorded again for 400 sec, corresponding to the “20 min” acquisition phase.

[0291] Stock solutions of test compounds were prepared at a concentration of 10 mM in DMSO (Sigma). 5-fold serial dilutions of the compounds were first prepared in DMSO. Compounds were then diluted in Assay buffer supplemented with 0.1% fatty-acid free bovine serum albumin (Sigma), reaching final compound concentrations of 128 pM to 10 .Math.M on the neurons. The Kv7 channel blocker XE-991 (Biotrend) was directly diluted in Assay buffer containing 0.1% fatty-acid free bovine serum albumin, yielding a final concentration of 10 .Math.M in the assay plate.

Analysis

[0292] Time-sequence data were exported using Screenworks® software (Molecular Devices) and converted with Orbit software (Idorsia Pharmaceuticals ltd.) to a format compatible with proprietary analysis softwares. A high-pass filter was then applied to flatten the signal using HTStudio (Idorsia Pharmaceuticals ltd.) to allow calculations of areas under the curve (AUC) for all time-point and compound concentrations. This allowed to calculate potencies (IC.sub.50) at both “Acute” phase and “20min” phase (“IC.sub.50acute” and “IC.sub.5020 min”) using IC50Studio (Idorsia Pharmaceuticals ltd.) as described hereafter. Note: alternatively, signal flattening and IC.sub.50 calculations can be achieved using commercially available softwares such as Igor Pro® from Wave Metric (“moving window” filter) and Prism 7.0 from GraphPad, respectively. [0293] “IC.sub.50acute”: the ratio of AUC fluorescence before and after compound addition was used to generate concentration-response curve (inhibition) using non-linear regression analysis with a 4-parameter fitting. [0294] “IC.sub.5020 min”: the AUC of fluorescence measured 20 min after compounds addition was used to generate concentration-response curve (inhibition) using non-linear regression analysis with a 4-parameter fitting.

[0295] IC.sub.50 value corresponds to the compound concentration that inhibits 50% of the neuronal oscillations in the presence of vehicle (top plateau). The maximum of inhibition corresponds to the full abolishment of oscillations (bottom plateau), which was obtained by addition of 100 .Math.M carbamazepine (Sigma). Shift value was calculated as follows: Shift value = (IC.sub.50acute value in presence of 10 .Math.M XE-991 [nM]) / (IC.sub.50acute value [nM]). If IC.sub.50 in presence of XE-991 could not be calculated, then the minimal Shift value was calculated as follows: Shift value= (highest tested concentration [nM]) / (IC.sub.50acute value [nM]), and the Shift value was annotated with “>”.

TABLE-US-00014 Rat oscillation IC.sub.50s and shift Example No FLIPR: IC.sub.50 [nM] Shift Example No FLIPR: IC.sub.50 [nM] Shift acute 20 min acute 20 min 1 486 855 >39 30 804 1477 4.9 2 291 466 10 31 190 763 >96 3 188 334 18 32 67 303 28 4 138 1060 25 33 946 1511 >4.9 5 296 216 8.6 34 247 291 13 6 126 271 39 35 231 717 13 7 63 100 21 36 245 344 15 8 168 331 3.9 37 26 32 32 9 766 1420 14 38 135 390 4.1 10 57 51 49 39 41 68 65 11 741 1365 10 40 149 248 29 12 154 164 >36 41 73 117 18 13 14 41 118 42 1173 1915 7.1 14 37 79 119 43 578 967 4.8 15 619 960 8.2 44 591 1350 12 16 716 848 5.6 45 131 701 6.7 17 202 239 16 46 148 257 15 18 377 1078 3.3 47 118 160 35 19 50 111 22 48 232 484 24 20 566 467 15 49 247 628 25 21 373 942 29 50 68 82 429 22 134 311 16 51 363 515 4.2 23 75 53 17 52 660 738 4.9 24 208 198 9.4 53 168 295 39 25 235 610 7.5 54 218 216 11 26 295 315 11 55 701 1163 2.4 27 1345 1727 8.4 56 123 547 32 28 117 203 47 57 146 314 13 29 367 2150 >4.6 58 51 105 236

C) Kv7.2/3 Assay (Performed at Charles River)

[0296] HEK293 cells were stably transfected with the appropriate ion channel cDNA(s) (human KCNQ2 and KCNQ3 genes). Cells were cultured in Dulbecco’s Modified Eagle Medium /Nutrient mixture F-12 (D-MEM/F-12) supplemented with 10% fetal bovine serum, 100 U/mL penicillin G sodium, 100 .Math.g/mL streptomycin sulfate and selection antibiotics. FLIPR Test Procedure: For FLIPR assay, cells were plated in 384-well black clear-bottomed microtiter plates (BD Biocoat Poly-D-Lysine Multiwell Cell Culture Plate) at 15′000 to 30′000 cells per well. Cells were incubated at 37° C. overnight or until cells reached sufficient density in the wells (near confluent monolayer) to use in fluorescence assays. Fluorescence changes triggered by agonist application were recorded using FLIPR® Tetra and displayed with Screenworks® 4.2 software (Molecular Devices). Assays were performed with the FLIPR potassium assay kit (Molecular Devices) according to the manufacturer’s instructions. Dye-loading: Growth media was removed and replaced with 20 .Math.L of dye loading buffer for 60 min at room temperature. FLIPR Recording (agonist mode): Stock solutions of test compounds were prepared at a concentration of 33.3 mM in DMSO. 5 .Math.L of 5x concentrated test, vehicle, or control compounds solutions prepared in the stimulation buffer (K.sup.+-free buffer with 5 mM TI.sup.+) were added to each well and fluorescence recording was continued for 5 min. The agonist effect (EC.sub.50 and % effect) of test or control compounds on Kv7 channels was determined as follows: Raw data was exported using Screenworks® 4.2 software and the fluorescence traces were analysed using Microsoft Excel (Microsoft Corp., Redmond, WA). The test compounds responses were expressed as % of maximum response of the control compound Flupirtine (Sigma-Aldrich), which was tested at concentrations ranging from 0.03 to 100 .Math.M. Concentration-response data were fitted to a Hill equation. Non-linear least squares fits were made assuming a simple binding model. If appropriate, fits were weighted by the standard deviation. No assumptions about the fit parameters were made; the fit parameters were determined by the algorithm.

TABLE-US-00015 Kv7.2/7.3 activation Example No FLIPR: EC.sub.50 (nM) Example No FLIPR: EC.sub.50 (nM) 24 143 47 59 37 30 53 82

III. Pharmacological Experiments

Formulation and Administration

[0297] Compounds were formulated in a 10% polyethylene glycol 400 (PEG 400) / 90% aqueous solution of 0.5 % methylcellulose (MC 0.5%). Firstly drugs are dissolved in PEG 400 and then suspended in MC 0.5 % for oral gavage at Xmg/5 mL/kg (X see table).

Audiogenic Seizure-Sensitive Mouse Model of Generalized Convulsive Seizures

[0298] 1. Procedure: Following two days of acclimatisation, auditory seizures are induced in male juvenile DBA/2J mice (22-24 days old; Janvier Labs, France). Each mouse is placed individually in the exposure chamber, an hemispheric acrylic glass dome (diameter: 50 cm) within a sound-attenuated box. The soundattenuated box is equipped with two house lights and a camera system (Fire-I from Unibrain) in order to observe and record the behavioral seizure response. After 60 seconds of habituation, the stimulus, a mixed frequency tone of 15-20 kHz at 110 dB (SASLab Lite, Avisoft Bioacoustics), is played from a speaker that is placed on the top center of the dome. The stimulus is applied for 60 seconds maximum or until the mouse shows tonic extension of the hind limbs. Seizures are classified as following: stage 0, normal behavior; stage 1, wild running; stage 2, generalized clonus; stage 3, tonic extension of the hind limbs. [0299] 2. Compounds testing: Acute compound effects on audiogenic generalized convulsive seizures are evaluated in independent groups of 8-10 mice randomly assigned. Following oral administration of compound or vehicle, the maximum seizure stage during sound exposure is assessed. Compounds are given 1 hour before exposure to the stimulus. Each mouse is exposed to the auditory stimulus only once and euthanized afterwards by CO.sub.2 inhalation.

TABLE-US-00016 Efficacies in the AGS mouse model Example No Formulation Dose and administration Time of challenge Seizure stage [% vs. vehicle] 23 90%MC0.5% + 10%PEG400 30 mg/kg po 1h -85 24 90%MC0.5% + 10%PEG400 30 mg/kg po 1h -100 37 90%MC0.5% + 10%PEG400 15 mg/kg po 1h -100 47 90%MC0.5% + 10%PEG400 10 mg/kg po 1h -92 53 90%MC0.5% + 10%PEG400 10 mg/kg po 1h -78

Amygdala-Kindling Rat Model

[0300] 1. Procedure: Adult male Wistar rats (Harlan Laboratories, Netherlands, or Charles Rivers, Germany; body weight 300-350 g) were stereotaxically implanted with twisted bipolar plastic-coated stainless steel electrode (MS333-2-BIU 10 mm, Plastics One) into the right basolateral amygdala under isoflurane anesthesia. To place the electrode, trepanations were made in the skull and the electrode was lowered into the right basolateral amygdala (from bregma: anteriorposterior (AP): -2.5 mm, medio-lateral (ML): -3.5 mm, dorso-ventral (DV): -8.6 mm; α=10.sup.0) and secured to the skull with screws and dental acrylate. After one week of recovery, they were handled daily and habituated over one week to the kindling setup. Kindling procedure: For a kindling session each rat was placed individually into a smooth acrylic plastic, round-bottomed bowl (0 36 cm, height 36 cm, BASi movement-responsive caging system) and its intracranial implanted electrode was connected to the stimulator (STG4008, Multichannel Systems GmbH) and the recording devices (PowerLab 8/35, ADInstruments Ltd) via a cable (335-340/3 (C), Plastics One). For the kindling procedure, each rat was exposed once daily to an electrical stimulation and the behavioral symptoms of the evoked seizure were observed and classified according to the modified Racine scale (stage 0, arrest, wet dog shakes, normal behaviour; stage 1, facial twitches: nose, lips, eyes; stage 2, chewing, head nodding; stage 3, forelimb clonus; stage 4, rearing, falling on forelimbs; stage 5, rearing, falling on side or back, rolling). The electrical stimulus consists of a 1s-train of 50 Hz square-wave biphasic pulses of 1-ms duration at an intensity of 400.Math.A (suprathreshold intensity). The stimulus was applied daily until each rat was fully kindled, i.e. it showed seizures of severity stage 4 and 5 upon electrical stimulation in at least ten consecutive kindling sessions. Data Scoring and analysis. The duration of electroencephalographic seizures (afterdischarge, AD) was recorded using LabChart7 Pro software (ADInstruments Ltd). Simultaneously, videos were recorded to evaluate seizure stage (SS). [0301] 2. Compound testing: Acute drug effects were evaluated in groups of 6-8 fully kindled rats in a randomized cross-over design with 48h between drug and vehicle applications. Following oral administration of drug or vehicle, drug testing included determination of the afterdischarge threshold (the minimal stimulation intensity necessary to evoke an afterdischarge (electroencephalographically measured neuronal hyper-synchronous activity with an amplitude 2-times higher than baseline amplitude and a frequency of 2:1 Hz) of at least 3 sec duration) and monitoring electroencephalographic and behavioral correlates of the evoked seizure at ADT (afterdischarge threshold), including AD duration and SS, by a experimenter blind to treatment assignment.

TABLE-US-00017 Efficacies in the rat kindling model Example No Formulation Dose and administration Time of challenge Seizure stage [% vs veh] AD duration [% vs veh] 47 90%MC0.5% + 10%PEG400 10 mg/kg po 1 h -70 -85 53 90%MC0.5% + 10%PEG400 30 mg/kg po 1 h -81 -93