HYDROXYALKYL-PIPERAZINE DERIVATIVES AS CXCR3 RECEPTOR MODULATORS

Abstract

The invention relates to compounds of Formula (I)

##STR00001##

wherein n, X, R.sup.1 and R.sup.2 are as described in the description; to pharmaceutically acceptable salts thereof, and to the use of such compounds as medicaments, especially as modulators of the CXCR3 receptor.

Claims

1. A compound of Formula (I) ##STR00009## wherein n represents the integer 1 or 2; X represents N or CH; R.sup.1 represents heteroaryl, wherein the heteroaryl is a 5- to 10-membered monocyclic or bicyclic aromatic ring comprising 1, 2 or 3 heteroatoms independently selected from oxygen, nitrogen or sulphur, and wherein the heteroaryl is independently unsubstituted or mono- or di-substituted, wherein the substituents are independently selected from (C.sub.1-4)alkyl; (C.sub.3-6)cycloalkyl; (C.sub.1-4)alkoxy; (C.sub.1-2)alkoxy-(C.sub.1-2)alkyl; (C.sub.1-2)alkyl-carbonyl; hydroxy-(C.sub.1-4)alkyl; halogen; (C.sub.1-2)fluoroalkyl; phenyl; or heteroaryl, wherein the heteroaryl is a 5- or 6-membered monocyclic aromatic ring comprising 1, 2 or 3 heteroatoms independently selected from oxygen, nitrogen or sulphur, and wherein the heteroaryl is independently unsubstituted or mono-substituted with (C1-4)alkyl; and R.sup.2 represents (C.sub.3-6)cycloalkyl, (C.sub.1-4)alkoxy or (C.sub.1-2)fluoroalkyl; or a salt thereof.

2. The compound according to claim 1, wherein n represents the integer 1 or 2; X represents N; R.sup.1 represents a 5-membered monocyclic heteroaryl group comprising 2 or 3 nitrogen atoms which is independently mono- or di-substituted, wherein the substituents are independently selected from (C.sub.1-4)alkyl, (C.sub.3-6)cycloalkyl, (C.sub.1-2)alkoxy-(C.sub.1-2)alkyl, hydroxy-(C.sub.1-4)alkyl or (C.sub.1-2)fluoroalkyl; or a 9-membered bicyclic aromatic ring comprising 1, 2 or 3 nitrogen atoms which is unsubstituted or mono-substituted with (C.sub.1-4)alkoxy or halogen; and R.sup.2 represents (C.sub.3-6)cycloalkyl, (C.sub.1-4)alkoxy or trifluoromethyl; or a salt thereof.

3. The compound according to claim 1, wherein R.sup.1 represents a 5-membered monocyclic heteroaryl group comprising 2 or 3 nitrogen atoms which is independently mono- or di-substituted, wherein the substituents are independently selected from (C.sub.1-4)alkyl, (C.sub.3-6)cycloalkyl or (C.sub.1-2)fluoroalkyl; or a salt thereof.

4. The compound according to claim 1, wherein R.sup.2 represents trifluoromethyl; or a salt thereof.

5. The compound according to claim 1, which is also a compound of Formula (I.sub.TA) ##STR00010## wherein n represents the integer 1 or 2; R.sup.1A represents hydrogen, (C.sub.1-4)alkyl, (C.sub.3-6)cycloalkyl, (C.sub.1-2)alkoxy-(C.sub.1-2)alkyl, (C.sub.1-2)alkyl-carbonyl, hydroxy-(C.sub.1-4)alkyl, (C.sub.1-2)fluoroalkyl, phenyl or heteroaryl, wherein the heteroaryl is a 5- or 6-membered monocyclic aromatic ring comprising 1, 2 or 3 heteroatoms independently selected from oxygen, nitrogen or sulphur, and wherein the heteroaryl is independently unsubstituted or mono-substituted with (C.sub.1-4)alkyl; R.sup.1B represents hydrogen or (C.sub.1-4)alkyl; and R.sup.2 represents (C.sub.3-6)cycloalkyl, (C.sub.1-4)alkoxy or (C.sub.1-2)fluoroalkyl; or a salt thereof.

6. The compound according to claim 5, wherein R.sup.1A represents (C.sub.1-4)alkyl, (C.sub.3-6)cycloalkyl, (C.sub.1-2)alkoxy-(C.sub.1-2)alkyl, hydroxy-(C.sub.1-4)alkyl or (C.sub.1-2)fluoroalkyl; or a salt thereof.

7. The compound according to claim 5, wherein R.sup.1A represents (C.sub.1-4)alkyl or (C.sub.3-6)cycloalkyl; or a salt thereof.

8. The compound according to claim 5, wherein R.sup.1B represents hydrogen; or a salt thereof.

9. The compound according to claim 5, wherein R.sup.1B represents methyl; or a salt thereof.

10. The compound according to claim 5, wherein R.sup.2 represents trifluoromethyl; or a salt thereof.

11. The compound according to claim 1, wherein the compound is: 2-(3-tert-Butyl-[1,2,4]triazol-1-yl)-1-{(S)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 2-(3,5-Dimethyl-[1,2,4]triazol-1-yl)-1-{(S)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 2-(3-Cyclopropyl-[1,2,4]triazol-1-yl)-1-{(S)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 1-{(S)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-isopropyl-[1,2,4]triazol-1-yl)-ethanone; 1-{(S)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-trifluoromethyl-[1,2,4]triazol-1-yl)-ethanone; 1-{(S)-4-[4-(2-Ethoxy-pyrimidin-5-yl)-2-trifluoromethyl-thiazol-5-yl]-2-hydroxymethyl-piperazin-1-yl}-2-(3-trifluoromethyl-[1,2,4]triazol-1-yl)-ethanone; 2-(3,5-Dimethyl-[1,2,4]triazol-1-yl)-1-{(S)-4-[4-(2-ethoxy-pyrimidin-5-yl)-2-trifluoromethyl-thiazol-5-yl]-2-hydroxymethyl-piperazin-1-yl}-ethanone; 1-{(S)-4-[4-(2-Cyclopropyl-pyrimidin-5-yl)-2-trifluoromethyl-thiazol-5-yl]-2-hydroxymethyl-piperazin-1-yl}-2-(3,5-dimethyl-[1,2,4]triazol-1-yl)-ethanone; 1-{(S)-4-[4-(2-Cyclopropyl-pyrimidin-5-yl)-2-trifluoromethyl-thiazol-5-yl]-2-hydroxymethyl-piperazin-1-yl}-2-(3-trifluoromethyl-[1,2,4]triazol-1-yl)-ethanone; 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(6-trifluoromethyl-pyridin-3-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-isopropyl-[1,2,4]triazol-1-yl)-ethanone; 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-isopropyl-[1,2,4]triazol-1-yl)-ethanone; 2-(3,5-Dimethyl-[1,2,4]triazol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 2-Benzoimidazol-1-yl-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 2-(5-Fluoro-indol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(5-methoxy-indol-1-yl)-ethanone; 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methyl-[1,2,4]triazol-1-yl)-ethanone; 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methoxymethyl-[1,2,4]triazol-1-yl)-ethanone; 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-[3-(5-methyl-[1,2,4]oxadiazol-3-yl)-[1,2,4]triazol-1-yl]-ethanone; 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-phenyl-[1,2,4]triazol-1-yl)-ethanone; 2-(3-Acetyl-[1,2,4]triazol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 2-[3-(1-Hydroxy-ethyl)-[1,2,4]triazol-1-yl]-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methyl-pyrazol-1-yl)-ethanone; 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-pyridin-2-yl-[1,2,4]triazol-1-yl)-ethanone; 2-(3-Ethyl-5-methyl-[1,2,4]triazol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 2-(5-Ethyl-3-methyl-[1,2,4]triazol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-imidazo[4,5-b]pyridin-3-yl-ethanone; 2-(3,5-Dimethyl-[1,2,4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 1-{(R)-2-(2-Hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-trifluoromethyl-[1,2,4]triazol-1-yl)-ethanone; 2-(3-Cyclobutyl-[1,2,4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 2-(3-tert-Butyl-[1,2,4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 2-(3-Cyclopropyl-[1,2,4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 2-(3-Ethyl-[1,2,4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 2-(5-Ethyl-[1,2,4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 1-{(R)-2-(2-Hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-isopropyl-[1,2,4]triazol-1-yl)-ethanone; 1-{(R)-2-(2-Hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methyl-[1,2,4]triazol-1-yl)-ethanone; 1-{(R)-2-(2-Hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(5-methyl-[1,2,4]triazol-1-yl)-ethanone; 1-{(R)-2-(2-Hydroxy-ethyl)-4-[2-trifuoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methoxymethyl-[1,2,4]triazol-1-yl)-ethanone; or 2-(3-Difluoromethyl-[1,2,4]triazol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; or a salt thereof.

12. The compound according to claim 1, wherein the compound is: 1-{(R)-2-(2-Hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methyl-pyrazol-1-yl)-ethanone; 2-(3-Ethyl-pyrazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 2-(3-Cyclopropyl-pyrazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; 1-{(S)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methyl-pyrazol-1-yl)-ethanone; 2-(3-Ethyl-pyrazol-1-yl)-1-{(S)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; or 2-(3-Cyclopropyl-pyrazol-1-yl)-1-{(S)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone; or a salt thereof.

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

14. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, formulated as a medicament.

15. A method of preventing or treating a disease comprising administering to a subject in need thereof, an effective amount of the compound according to claim 1, wherein the disease is an autoimmune disorder, inflammatory disease, infectious disease, transplant rejection, fibrosis, neurodegenerative disorder, or cancer.

16. A method of preventing or treating a disease comprising administering to a subject in need thereof, an effective amount of the pharmaceutical composition according to claim 13, wherein the disease is an autoimmune disorder, inflammatory disease, infectious disease, transplant rejection, fibrosis, neurodegenerative disorder, or cancer.

Description

EXPERIMENTAL SECTION

[0137] Abbreviations (as used herein and in the description above): [0138] aq. aqueous [0139] Boc tert.-butyloxycarbonyl [0140] BSA Bovine serum albumine [0141] Bu butyl [0142] CC column chromatography on silica gel [0143] CHO Chinese hamster ovary [0144] CV column volume [0145] d day(s) [0146] DCM dichloromethane [0147] DEA diethylamine [0148] DIPEA N-ethyldiisopropylamine [0149] DMF dimethylformamide [0150] DMSO dimethylsulfoxide [0151] EA ethyl acetate [0152] EDTA ethylenediaminetetraacetic acid [0153] EGTA ethylene glycol tetraacetic acid [0154] Et ethyl [0155] FBS fetal bovine serum [0156] FLIPR Fluorescent imaging plate reader [0157] Fluo-4-AM 2-{[2-(2-{5-[bis(carboxymethyl)amino]-2-methylphenoxy}ethoxy)-4-(2,7-difluoro-6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl](carboxymethyl)amino}acetic acid [0158] G418 (2R,3S,4R,5R,6S)-5-amino-6-[(1R,2S,3S,4R,6S)-4,6-diamino-3-[(2R,3R,4R,5R)-3,5-dihydroxy-5-methyl-4-methylaminooxan-2-yl]oxy-2-hydroxycyclohexyl]oxy-2-(1-hydroxyethyl)oxane-3,4-diol [0159] h hour(s) [0160] HATU 2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium [0161] Hep heptanes [0162] HEPES 4-(2-hydroxyethyl)-piperazine-1-ethanesulfonic acid [0163] HV High vacuum [0164] HPLC high performance liquid chromatography [0165] iPr iso-propyl [0166] LC liquid chromatography [0167] m multiplet [0168] M molarity [mol L.sup.−1] [0169] Me methyl [0170] MS mass spectrometry [0171] min minute(s) [0172] NMR nuclear magnetic resonance spectroscopy [0173] org. organic [0174] PBS Phosphate buffered saline [0175] Pd/C palladium on carbon [0176] PG protecting group [0177] Ph phenyl [0178] Prep preparative [0179] rpm rotations per minute [0180] RT room temperature [0181] s singulet [0182] sat. Saturated [0183] sec second(s) [0184] TFA trifluoroacetic acid [0185] THF tetrahydrofuran [0186] TLC Thin layer chromatography [0187] t.sub.R retention time [0188] UPLC Ultra performance liquid chromatography

[0189] I. Chemistry

[0190] The following examples illustrate the preparation of biologically active compounds of the invention but do not at all limit the scope thereof.

[0191] General: All temperatures are stated in degrees Celsius (° C.). Unless otherwise indicated, the reactions take place at RT under an argon atmosphere and are run in a flame dried round-bottomed flask equipped with a magnetic stir bar.

[0192] Characterization Methods Used:

[0193] The LC-MS retention times have been obtained using the following elution conditions: I) LC-MS (A):

[0194] Zorbax SB-Aq, 3.5 μm, 4.6×50 mm column thermostated at 40° C. The two elution solvents were as follows: solvent A=water+0.04% TFA; solvent B=MeCN. 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-00001 t (min) 0 1.0 1.45 1.55 Solvent A (%) 95 5 5 95 Solvent B (%) 5 95 95 5

[0195] II) LC-MS (B):

[0196] Acquity UPLC HSS T3 C18 1.8 μm 2.1×50 mm ID column from Waters, thermostated in the Acquity UPLC Column Manager (60° C.) was used. The two elution solvents were as follows: solvent A=water+0.05% formic acid; solvent B=MeCN+0.045% formic acid. The eluent flow rate was 1 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 1.4 1.8 1.9 2.0 Solvent A (%) 98 5 2 2 98 Solvent B (%) 2 95 98 98 2

[0197] III) LC-MS (C):

[0198] Acquity UPLC CSH C18 1.7 m 2.1×50 mm ID column from Waters, thermostated in the Acquity UPLC Column Manager (60° C.) was used. The two elution solvents were as follows: solvent A=water+0.05% formic acid; solvent B=MeCN+0.045% formic acid. The eluent flow rate was 1 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 1.4 1.8 1.9 2.0 Solvent A (%) 98 5 2 2 98 Solvent B (%) 2 95 98 98 2

[0199] Compound purity and identity was further confirmed by NMR spectroscopy (Bruker Avance II 400 MHz Ultrashield™ or Bruker Ascend™ 500 equipped with a 5 mm DCH cryoprobe), 1H (400 MHz or 500 MHz), 19F (376 MHz). The chemical shifts are reported in parts per million (ppm) relative to tetramethylsilane (TMS) or trichlorofluoromethane, and multiplicities are given as s (singlet) or m (multiplet).

[0200] Preparative LC-MS Methods Used:

[0201] The purifications by preparative LC-MS have been performed using the conditions described hereafter.

[0202] I) Preparative LC-MS (I):

[0203] A X-Bridge column (Waters C18, 10 μm OBD, 30×75 mm) was used. The two elution solvents were as follows: solvent A=water+0.5% NH.sub.4OH (25%); solvent B=MeCN. The eluent flow rate was 75 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 tables below (a linear gradient being used between two consecutive time points):

TABLE-US-00004 t (min) 0 0.01 4.0 6.0 6.2 6.6 Solvent A (%) 80 80 5 5 80 80 Solvent B (%) 20 20 95 95 20 20

[0204] II) Preparative LC-MS (II):

[0205] A X-Bridge column (Waters C18, 10 μm OBD, 30×75 mm) was used. The two elution solvents were as follows: solvent A=water+0.5% NH.sub.4OH (25%); solvent B=MeCN. The eluent flow rate was 75 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 tables below (a linear gradient being used between two consecutive time points):

TABLE-US-00005 t (min) 0 0.01 4.0 6.0 6.2 6.6 Solvent A (%) 90 90 5 5 90 90 Solvent B (%) 10 10 95 95 10 10

[0206] III) Preparative LC-MS (III):

[0207] An Atlantis column (Waters T3, 10 μm OBD, 30×75 mm) was used. The two elution solvents were as follows: solvent A=water+0.5% formic acid; solvent B=MeCN. The eluent flow rate was 75 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 tables below (a linear gradient being used between two consecutive time points):

TABLE-US-00006 t (min) 0 0.01 3.5 6.0 6.2 6.6 Solvent A (%) 70 70 5 5 70 70 Solvent B (%) 30 30 95 95 30 30

[0208] IV) Preparative LC-MS (IV):

[0209] An Atlantis column (Waters T3, 10 μm OBD, 30×75 mm) was used. The two elution solvents were as follows: solvent A=water+0.5% formic acid; solvent B=MeCN. The eluent flow rate was 75 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 tables below (a linear gradient being used between two consecutive time points):

TABLE-US-00007 t (min) 0 0.01 4.0 6.0 6.2 6.6 Solvent A (%) 80 80 5 5 80 80 Solvent B (%) 20 20 95 95 20 20

[0210] V) Preparative LC-MS (V):

[0211] X-Bridge column (Waters C18, 10 μm OBD, 30×75 mm) was used. The two elution solvents were as follows: solvent A=water+0.5% NH.sub.4OH (25%); solvent B=MeCN. The eluent flow rate was 75 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 tables below (a linear gradient being used between two consecutive time points):

TABLE-US-00008 t (min) 0 0.01 3.5 6.0 6.2 6.6 Solvent A (%) 70 70 5 5 70 70 Solvent B (%) 30 30 95 95 30 30

[0212] VI) Preparative LC-MS (VI):

[0213] An Atlantis column (Waters T3, 10 μm OBD, 30×75 mm) was used. The two elution solvents were as follows: solvent A=water+0.5% formic acid; solvent B=MeCN. The eluent flow rate was 75 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 tables below (a linear gradient being used between two consecutive time points):

TABLE-US-00009 t (min) 0 0.01 3 4.0 6.0 6.2 6.6 Solvent A (%) 95 95 50 5 5 95 95 Solvent B (%) 5 5 50 95 95 5 5

[0214] Preparative Chiral HPLC Methods Used:

[0215] The purifications by preparative chiral HPLC have been performed using the conditions described hereafter.

[0216] I) Preparative Chiral HPLC (I):

[0217] A ChiralPak IB column (5 m, 30×250 mm) was used. The elution solvent was Hep/EtOH/DEA 50/50/0.1, run for 10 min and at a flow rate of 16 mL/min.

[0218] II) Preparative Chiral HPLC (11):

[0219] A ChiralPak IF column (5 m, 20×250 mm) was used. The elution solvent was Hep/EtOH 50/50, run for 8.7 min and at a flow rate of 19 mL/min.

[0220] III) Preparative Chiral HPLC (111):

[0221] A (R,R) Whelk-01 column (10 μm, 50×250 mm) was used. The elution solvent was Hep/EtOH 70/30, run for 16.3 min and at a flow rate of 100 mL/min.

[0222] IV) Preparative Chiral HPLC (IV):

[0223] A ChiralPak IB column (5 μm, 30×250 mm) was used. The elution solvent was Hep/EtOH 70/30, run for 11.8 min and at a flow rate of 34 mL/min.

[0224] V) Preparative Chiral HPLC (V):

[0225] A ChiralPak IB column (5 m, 30×250 mm) was used. The elution solvent was Hep/EtOH 50/50, run for 7.6 min and at a flow rate of 34 mL/min.

[0226] VI) Preparative Chiral HPLC (VI):

[0227] A ChiralPak IB column (5 m, 30×250 mm) was used. The elution solvent was Hep/EtOH 60/40, run for 9 min and at a flow rate of 40 mL/min.

[0228] VII) Preparative Chiral HPLC (VII):

[0229] A ChiralPak IC column (5 m, 30×250 mm) was used. The elution solvent was Hep/EtOH 70/30, run for 12 min and at a flow rate of 34 mL/min.

Example 1: 2-(3-tert-Butyl-[1,2,4]triazol-1-yl)-1-{(S)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

1.1. 5-Bromo-2-trifluoromethyl-thiazole-4-carboxylic acid

[0230] To a solution of 2-(trifluoromethyl)thiazole-4-carboxylic acid (3.2 g) in anhydrous THF (60 mL) under argon cooled down to −78° C. was added n-BuLi (1.6M in hexane, 21.3 mL) dropwise over 15 min so that the internal temperature did not rise above −60° C. A solution of Br.sub.2 (0.92 mL) in cyclohexane (8 mL) was then added dropwise to keep the internal temperature below −60° C. The resulting mixture was stirred at −78° C. for 2 h and carefully quenched by addition of water (50 mL). Citric acid (10%) was added until pH=2 and the mixture was extracted with EA. The org. layers were washed with brine, dried (MgSO.sub.4), filtered off and evaporated to dryness to afford 4.15 g of brown solid, used without further purification. LC-MS (A): t.sub.R=0.67 min. F-NMR (CD.sub.3OD): −63.57 ppm (s).

1.2. 5-Bromo-2-trifluoromethyl-thiazole-4-carboxylic acid methyl ester

[0231] To a solution of intermediate 1.1 (12 g), MeOH (130 mL) was added H.sub.2SO.sub.4 (96%, 6.5 mL) and the mixture stirred at 70° C. for 3 h. After cooling down, the reaction mixture was quenched with sat. aq. Na.sub.2CO.sub.3 and the solvent partially evaporated off. The residue was diluted with DCM and washed with aq. sat. Na.sub.2CO.sub.3 (1×), water (1×) and brine (1×), and the aq. phases were extracted with DCM (2×). The combined org. layers were dried over MgSO.sub.4, filtrated off, evaporated and dried under HV to afford 12 g of brown resin. LC-MS (A): t.sub.R=0.83 min. F-NMR (CD.sub.3OD): −63.59 ppm (s).

1.3. (S)-2-Hydroxymethyl-4-(4-methoxycarbonyl-2-trifluoromethyl-thiazol-5-yl)-piperazine-1-carboxylic acid tert-butyl ester

[0232] To a solution of intermediate 1.2 (4 g) in MeCN (100 mL) were added (S)-1-Boc-2-hydroxymethylpiperazine (3.07 g) and DIPEA (3.54 mL) at RT. The reaction mixture was stirred at 80° C. for 28 h. After cooling down, the reaction mixture was diluted with EA and washed with water (2×) and brine. The aq. layers were extracted with EA. The combined org. layers were dried over MgSO.sub.4, filtrated off and evaporated to dryness. The crude was purified by CC (Biotage, SNAP 340 g, solvent A: Hep; solvent B: EA; gradient in % B: 30 over 3CV, 30 to 50 over 5CV, 50 over 3CV) to afford 4 g of yellow foam. LC-MS (A): t.sub.R=0.87 min; [M+H].sup.+: 426.0.

1.4. (S)-4-(4-Carboxy-2-trifluoromethyl-thiazol-5-yl)-2-hydroxymethyl-piperazine-1-carboxylic acid tert-butyl ester

[0233] To a solution of intermediate 1.3 (3.96 g) in EtOH (20 mL) was added 1M NaOH (20 mL) at RT and the reaction mixture was stirred for 1 h10. The solvent was evaporated off and the residue acidified to pH 2-3 by the addition of aq. citric acid (10%). The aq. layer was extracted with DCM (3×) and the combined org. layers were dried over MgSO.sub.4 and concentrated to dryness to afford 2.89 g as beige solid. LC-MS (A): t.sub.R=0.79 min; [M+H].sup.+: 412.1.

1.5. (S)-4-(4-Bromo-2-trifluoromethyl-thiazol-5-yl)-2-hydroxymethyl-piperazine-1-carboxylic acid tert-butyl ester

[0234] To a solution of intermediate 1.4 (2.88 g) in THF (55 mL) were added LiBr (614 mg) and (diacetoxyiodo)benzene (2.3 g) at RT. The resulting suspension was stirred at RT overnight. The reaction mixture was evaporated off and the residue taken up with H.sub.2O/DCM and extracted with DCM (3×). The combined org. layers were dried over MgSO.sub.4, filtrated off and evaporated to dryness. The crude was purified by CC (Biotage, SNAP 100 g cartridge, solvent A: Hep; solvent B: EA; gradient in % B: 10 for 5CV, 10 to 30 over 5CV, 40 for 3CV) to afford 2.5 g as white solid. LC-MS (A): t.sub.R=0.93 min; [M+H].sup.+: 445.9.

1.6. (S)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazine-1-carboxylic acid tert-butyl ester

[0235] A mixture of intermediate 1.5 (2.3 g), 2-(trifluoromethyl)pyrimidine-5-boronic acid (1.65 g), Pd(PPh.sub.3).sub.2Cl.sub.2 (202 mg), 1M Na.sub.2CO.sub.3 (15 mL) in MeCN (15 mL) was vigorously stirred at 80° C. under argon for 20 h. The reaction mixture was allowed to cool down to RT, diluted with H.sub.2O and extracted with DCM (3×). The combined org. layers were dried over MgSO.sub.4, filtrated off and evaporated to dryness. The crude was purified by CC (Biotage, SNAP 340 g cartridge, solvent A: Hep; solvent B: EA; gradient in % B: 10 for 5CV, 10 to 30 over 6CV, 30 for 3CV) to afford 2.22 g as yellow foam. LC-MS (A): t.sub.R=0.98 min; [M+H].sup.+: 513.9.

1.7. {(S)-4-[2-Trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-2-yl}-methanol; as hydrochloride salt

[0236] A mixture of intermediate 1.6 (2.25 g) in HCl (22 mL, 4M in dioxane) was stirred at RT for 1 h. The reaction mixture was evaporated and dried under HV to give 2.18 g as brown foam. LC-MS (A): t.sub.R=0.67 min; [M+H.sup.++CH.sub.3CN].sup.+: 413.2.

1.8. (3-tert-Butyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester

[0237] To a solution of 3-tert-butyl-1H-1,2,4-triazole (225 mg) in MeCN (12 mL) was added Cs.sub.2C03 (586 mg) followed by benzyl bromoacetate (0.29 mL) and the mixture was stirred at RT for 1 h45. The reaction mixture was diluted with EA and washed with water (2×) and brine. The aq. layers were extracted with EA (2×) and the combined org. layers were dried over MgSO.sub.4, filtered off and evaporated to dryness. The residue was purified by CC (Biotage, SNAP 25 g cartridge, solvent A: DCM; solvent B: DCM/MeOH 8:2; gradient in % B: 15 for 8CV) to afford 216 mg as yellowish solid (single regioisomer). LC-MS (A): t.sub.R=0.73 min; [M+H].sup.+: 274.1.

1.9. (3-tert-Butyl-[1, 2, 4]triazol-1-yl)-acetic acid

[0238] A flask containing intermediate 1.8 (202 mg), Pd/C (39 mg) in EtOH (2 mL) was evacuated and backfilled with argon (3×), afterwards evacuated and backfilled with H.sub.2 (3×) and the reaction mixture stirred at RT for 2 h. The reaction mixture was filtered over a celite plug and the filtrate was evaporated to dryness to afford 130 mg as white solid. LC-MS (A): t.sub.R=0.36 min; [M+H].sup.+: 184.3.

1.10. 2-(3-tert-Butyl-[1, 2, 4]triazol-1-yl)-1-{(S)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0239] A mixture of intermediate 1.7 (30 mg), intermediate 1.9 (11 mg), HATU (26 mg), and NEt.sub.3 (26 μL) in DCM (1.5 mL) was stirred at RT overnight. The reaction mixture was evaporated to dryness and the crude was purified by Prep LC-MS (IV) to afford 6 mg as white solid. LC-MS (C): t.sub.R=1.12 min; [M+H].sup.+: 579.2.

Example 2: 2-(3,5-Dimethyl-[1,2,4]triazol-1-yl)-1-{(S)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0240] To a solution of intermediate 1.7 (2.15 g) in DCM (66 mL) were added (3,5-dimethyl-1H-1,2,4-triazole-1-yl)acetic acid (743 mg), DIPEA (2.05 mL) and HATU (2.37 g) at RT and the mixture was stirred at RT for 3 h. The reaction mixture was diluted with DCM and washed with 1M NaHSO.sub.4, aq. sat. NaHCO.sub.3 and brine. The aq. layers were re-extracted with 2×DCM and the combined org. layers were dried over MgSO.sub.4, filtrated off and evaporated to dryness. Purification by CC (Biotage, SNAP 100 g cartridge, solvent A: DCM; solvent B: DCM/MeOH 8:2+0.1% Et.sub.3N; gradient in % B: 15 for 5CV, 15 to 25 over 3CV, 25 for 5CV). Residual DIPEA in the product was removed by extraction in DCM/H.sub.2O to give 778 mg as white solid.

[0241] LC-MS (C): t.sub.R=0.97 min; [M+H].sup.+: 551.1.

Example 3: 2-(3-Cyclopropyl-[1,2,4]triazol-1-yl)-1-{(S)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

3. 1. (3-Cyclopropyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester

[0242] To a solution of 3-cyclopropyl-1H-1,2,4-triazole (500 mg) in THF (10 mL) was added NaH (60% in oil, 174 mg) at 0° C. and the mixture was stirred for 5 min. Benzyl bromoacetate (0.719 mL) was then added and the reaction mixture was stirred for 45 min at 0° C. The mixture was quenched with sat. aq. NH.sub.4Cl and the aq. layer was extracted with EA (3×). The combined org. layers were dried over MgSO.sub.4, filtrated off and evaporated to dryness. Purification by CC (Biotage, SNAP 25 g cartridge, solvent A: DCM; solvent B: MeOH; gradient in % B: 1 for 4CV, 1 to 10 over 10 CV, 10 for 2CV) afforded 831 mg as a mixture of regioisomers. Separation by preparative chiral HPLC (II) afforded two regioisomers:

[0243] First eluting fraction: (5-cyclopropyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester. LC-MS (A): t.sub.R=0.75 min; [M+H].sup.+: 258.1. Roesy signal seen between CH.sub.2CO.sub.2 at 5.04 ppm and CH (cyclopropyl) at 1.74 ppm.

[0244] Second eluting fraction: (3-cyclopropyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester. LC-MS (A): t.sub.R=0.75 min; [M+H].sup.+: 258.1. Roesy signal seen between CH.sub.2CO.sub.2 at 4.94 ppm and CH (triazole) at 8.08 ppm.

3.2. (3-Cyclopropyl-[1,2, 4]triazol-1-yl)-acetic acid

[0245] This compound was prepared using a method analogous to that of Example 1, step 1.9, intermediate 3.1 ((3-cyclopropyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester) replacing intermediate 1.8. LC-MS (A): t.sub.R=0.33 min; [M+H].sup.+: 168.47.

3.3. 2-(3-Cyclopropyl-[1, 2, 4]triazol-1-yl)-1-{(S)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0246] This compound was prepared using a method analogous to that of Example 1, step 1.10, intermediate 3.2 replacing intermediate 1.9. The crude was purified by Prep LC-MS (IV). LC-MS (C): t.sub.R=1.05 min; [M+H].sup.+: 563.1.

Example 4: 1-{(S)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-isopropyl-[1,2,4]triazol-1-yl)-ethanone

4. 1. (3-Isopropyl-[1, 2, 4]triazol-1-yl)-acetic acid benzyl ester

[0247] This compound was prepared using a method analogous to that of Example 1, step 1.8, 3-isopropyl-1H-1,2,4-triazole replacing 3-tert-butyl-1H-1,2,4-triazole. The mixture of regioisomers was purified by preparative chiral HPLC (Ill). First eluting fraction: (3-isopropyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester. LC-MS (A): t.sub.R=0.76 min; [M+H].sup.+: 260.2. Roesy signal seen between CH.sub.2 at 4.96 ppm and CH (triazole) at 8.08 ppm.

[0248] Second eluting fraction: (5-isopropyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester. LC-MS (A): t.sub.R=0.76 min; [M+H].sup.+: 260.2. Roesy signal seen between CH.sub.2 at 4.96 ppm and CH (isopropyl) at 2.97 ppm.

4.2. (3-Isopropyl-[1,2,4]triazol-1-yl)-acetic acid

[0249] This compound was prepared using a method analogous to that of Example 1, step 1.9, (3-isopropyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester from step 4.1 replacing intermediate 1.8. LC-MS (A): t.sub.R=0.30 min; [M+H].sup.+: 170.2.

4.3. 1-{(S)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-isopropyl-[1, 2, 4]triazol-1-yl)-ethanone

[0250] This compound was prepared using a method analogous to that of Example 1, step 1.10, intermediate 4.2 replacing intermediate 1.9. The crude was purified by Prep LC-MS (IV). LC-MS (C): t.sub.R=1.07 min; [M+H].sup.+: 564.9.

Example 5: 1-{(S)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-trifluoromethyl-[1,2,4]triazol-1-yl)-ethanone

5.1. (3-Trifluoromethyl-[1, 2, 4]triazol-1-yl)-acetic acid benzyl ester

[0251] This compound was prepared using a method analogous to that of Example 1, step 1.8, 3-(trifluoromethyl)-1H-1,2,4-triazole replacing 3-tert-butyl-1H-1,2,4-triazole. The desired compound was obtained after CC as single regioisomer. LC-MS (A): t.sub.R=0.85 min; [M+H].sup.+: 286.1.

5.2. (3-Trifluoromethyl-[1, 2, 4]triazol-1-yl)-acetic acid

[0252] This compound was prepared using a method analogous to that of Example 1, step 1.9, intermediate 5.1 replacing intermediate 1.8. LC-MS (A): t.sub.R=0.42 min; [M+H].sup.+: 196.1.

5.3. 1-{(S)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-trifluoromethyl-[1, 2, 4]triazol-1-yl)-ethanone

[0253] This compound was prepared using a method analogous to that of Example 1, step 1.10, intermediate 5.2 replacing intermediate 1.9. Additional 1.1 eq of HATU and 1 eq of intermediate 5.2 were added and the reaction mixture was further stirred for 24 h. The crude was purified by Prep LC-MS (IV). LC-MS (C): t.sub.R=1.15 min; [M+H].sup.+: 591.1.

Example 6: 1-{(S)-4-[4-(2-Ethoxy-pyrimidin-5-yl)-2-trifluoromethyl-thiazol-5-yl]-2-hydroxymethyl-piperazin-1-yl}-2-(3-trifluoromethyl-[1,2,4]triazol-1-yl)-ethanone

6.1. (S)-4-[4-(2-Ethoxy-pyrimidin-5-yl)-2-trifluoromethyl-thiazol-5-yl]-2-hydroxymethyl-piperazine-1-carboxylic acid tert-butyl ester

[0254] This compound was prepared using a method analogous to that of Example 1, step 1.6, 2-ethoxypyrimidine-5-boronic acid replacing 2-(trifluoromethyl)pyrimidine-5-boronic acid. LC-MS (A): t.sub.R=0.92 min; [M+H].sup.+: 490.2.

6.2. (S)-(4-(4-(2-Ethoxypyrimidin-5-yl)-2-(trifluoromethyl) thiazol-5-yl)piperazin-2-yl)methanol, as hydrochloride salt

[0255] This compound was prepared using a method analogous to that of Example 1, step 1.7, intermediate 6.1 replacing intermediate 1.6. LC-MS (A): t.sub.R=0.57 min; [M+H].sup.+: 390.2.

6.3. 1-{(S)-4-[4-(2-Ethoxy-pyrimidin-5-yl)-2-trifluoromethyl-thiazol-5-yl]-2-hydroxymethyl-piperazin-1-yl}-2-(3-trifluoromethyl-[1, 2, 4]triazol-1-yl)-ethanone

[0256] This compound was prepared using a method analogous to that of Example 1, step 1.10, intermediate 6.2 replacing intermediate 1.7, intermediate 5.2 replacing intermediate 1.9 and DMF replacing DCM. The crude was purified by Prep LC-MS (I). LC-MS (C): t.sub.R=1.09 min; [M+H].sup.+: 567.1.

Example 7: 2-(3,5-Dimethyl-[1,2,4]triazol-1-yl)-1-{(S)-4-[4-(2-ethoxy-pyrimidin-5-yl)-2-trifluoromethyl-thiazol-5-yl]-2-hydroxymethyl-piperazin-1-yl}-ethanone

[0257] This compound was prepared using a method analogous to that of Example 6, step 6.3, 3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid replacing intermediate 5.2. The crude was purified by Prep LC-MS (I). LC-MS (B): t.sub.R=1.28 min; [M+H].sup.+: 527.2.

Example 8: 1-{(S)-4-[4-(2-Cyclopropyl-pyrimidin-5-yl)-2-trifluoromethyl-thiazol-5-yl]-2-hydroxymethyl-piperazin-1-yl}-2-(3,5-di methyl-[1,2,4]triazol-1-yl)-ethanone

8.1. (S)-4-[4-(2-Cyclopropyl-pyrimidin-5-yl)-2-trifluoromethyl-thiazol-5-yl]-2-hydroxymethyl-piperazine-1-carboxylic acid tert-butyl ester

[0258] This compound was prepared using a method analogous to that of Example 1, step 1.6, 2-cyclopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin replacing 2-(trifluoromethyl)pyrimidine-5-boronic acid. LC-MS (A): t.sub.R=0.92 min; [M+H].sup.+: 486.2.

8.2. (S)-(4-(4-(2-Cyclopropylpyrimidin-5-yl)-2-(trifluoromethyl)thiazol-5-yl)piperazin-2-yl)methanol, as hydrochloride salt

[0259] This compound was prepared using a method analogous to that of Example 1, step 1.7, intermediate 8.1 replacing intermediate 1.6. LC-MS (A): t.sub.R=0.57 min; [M+H].sup.+: 386.1.

8.3. 1-{(S)-4-[4-(2-Cyclopropyl-pyrimidin-5-yl)-2-trifluoromethyl-thiazol-5-yl]-2-hydroxymethyl-piperazin-1-yl}-2-(3, 5-dimethyl-[1, 2, 4]triazol-1-yl)-ethanone

[0260] This compound was prepared using a method analogous to that of Example 7, intermediate 8.2 replacing intermediate 6.2. The crude was purified by Prep LC-MS (I). LC-MS (C): t.sub.R=0.9 min; [M+H].sup.+: 523.2.

Example 9: 1-{(S)-4-[4-(2-Cyclopropyl-pyrimidin-5-yl)-2-trifluoromethyl-thiazol-5-yl]-2-hydroxymethyl-piperazin-1-yl}-2-(3-trifluoromethyl-[1,2,4]triazol-1-yl)-ethanone

[0261] This compound was prepared using a method analogous to that of Example 8, step 8.3, intermediate 5.2 replacing (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid. The crude was purified by Prep LC-MS (I) followed by Prep TLC (DCM/MeOH 97:3). LC-MS (C): t.sub.R=1.1 min; [M+H].sup.+: 563.1.

Example 10: 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(6-trifluoromethyl-pyridin-3-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-isopropyl-[1,2,4]triazol-1-yl)-ethanone

10.1. (R)-4-(4-Bromo-2-trifluoromethyl-thiazol-5-yl)-2-hydroxymethyl-piperazine-1-carboxylic acid tert-butyl ester

[0262] This compound was prepared using a method analogous to that of Example 1, following steps 1.1 to steps 1.5, (R)-1-Boc-2-hydroxymethylpiperazine replacing (S)-1-Boc-2-hydroxymethylpiperazine in step 1.3. LC-MS (A): t.sub.R=0.92 min; [M+H].sup.+: 445.9.

10.2. (R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(6-trifluoromethyl-pyridin-3-yl)-thiazol-5-yl]-piperazine-1-carboxylic acid tert-butyl ester

[0263] This compound was prepared using a method analogous to that of Example 1, step 1.6, 2-(trifluoromethyl)pyridine-5-boronic acid replacing 2-(trifluoromethyl)pyrimidine-5-boronic acid. LC-MS (A): t.sub.R=0.99 min; [M+H].sup.+: 512.9.

10.3. (R)-(4-(2-(trifluoromethyl)-4-(6-(trifluoromethyl)pyridin-3-yl)thiazol-5-yl)piperazin-2-yl)methanol, as hydrochloride salt

[0264] This compound was prepared using a method analogous to that of Example 1, step 1.7, intermediate 10.2 replacing intermediate 1.6. LC-MS (A): t.sub.R=0.68 min; [M+H].sup.+: 413.1.

10.4. 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(6-trifluoromethyl-pyridin-3-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-isopropyl-[1, 2, 4]triazol-1-yl)-ethanone

[0265] This compound was prepared using a method analogous to that of Example 4, step 4.3, intermediate 10.3 replacing intermediate 1.7 and DMF replacing DCM. The crude was purified by Prep LC-MS (I). LC-MS (C): t.sub.R=1.1 min; [M+H].sup.+: 564.2.

Example 11: 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-isopropyl-[1,2,4]triazol-1-yl)-ethanone

11.1. (R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazine-1-carboxylic acid tert-butyl ester

[0266] This compound was prepared using a method analogous to that of Example 10, step 10.2, 2-(trifluoromethyl)pyrimidine-5-boronic acid replacing 2-(trifluoromethyl)pyridine-5-boronic acid. LC-MS (A): t.sub.R=0.98 min; [M+H].sup.+: 513.9.

11.2. (R)-(4-(2-(trifluoromethyl)-4-(2-(trifluoromethyl)pyrimidin-5-yl)thiazol-5-yl) piperazin-2-yl)methanol, as hydrochloride salt

[0267] This compound was prepared using a method analogous to that of Example 10, step 10.3, intermediate 11.1 replacing intermediate 10.2. LC-MS (A): t.sub.R=0.67 min; [M+H].sup.+: 414.0.

11.3. 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-isopropyl-[1, 2, 4]triazol-1-yl)-ethanone

[0268] This compound was prepared using a method analogous to that of Example 10, step 10.4, intermediate 11.2 replacing intermediate 10.3. The crude was purified by Prep LC-MS (I). LC-MS (C): t.sub.R=1.07 min; [M+H].sup.+: 565.4.

Example 12: 2-(3,5-Dimethyl-[1,2,4]triazol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0269] This compound was prepared using a method analogous to that of Example 11, step 11.3, (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid replacing intermediate 4.2. The crude was purified by Prep LC-MS (11). LC-MS (C): t.sub.R=0.97 min; [M+H].sup.+: 551.1.

Example 13: 2-Benzoimidazol-1-yl-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0270] This compound was prepared using a method analogous to that of Example 11, step 11.3, 2-(1H-benzimidazol-1-yl)acetic acid replacing intermediate 4.2. The crude was purified by Prep LC-MS (I). LC-MS (C): t.sub.R=0.89 min; [M+H].sup.+: 572.1.

Example 14: 2-(5-Fluoro-indol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0271] This compound was prepared using a method analogous to that of Example 11, step 11.3, (5-fluoroindol-1-yl)acetic acid replacing intermediate 4.2. The crude was purified by Prep LC-MS (V). LC-MS (C): t.sub.R=1.25 min; [M+H].sup.+: 589.1.

Example 15: 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(5-methoxy-indol-1-yl)-ethanone

[0272] This compound was prepared using a method analogous to that of Example 11, step 11.3, (5-methoxy-1H-indol-1-yl)acetic acid replacing intermediate 4.2. The crude was purified by Prep LC-MS (I). LC-MS (C): t.sub.R=1.22 min; [M+H].sup.+: 601.1.

Example 16: 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methyl-[1,2,4]triazol-1-yl)-ethanone

16.1. (3-Methyl-[1, 2, 4]triazol-1-yl)-acetic acid benzyl ester

[0273] This compound was prepared using a method analogous to that of Example 1, step 1.8, 3-methyl-1H-1,2,4-triazole replacing 3-tert-butyl-1H-1,2,4-triazole. The mixture of regioisomers was purified by preparative chiral HPLC (IV). First eluting fraction: (5-methyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester. LC-MS (A): t.sub.R=0.68 min; [M+H].sup.+: 232.16. .sup.1H-NMR (CDCl.sub.3): 7.83 (s, 1H); 7.40-7.33 (m, 5H); 5.23 (s, 2H); 4.93 (s, 2H); 2.43 (s, 3H). Roesy signal seen between CH.sub.2 at 4.93 ppm and CH.sub.3 at 2.43 ppm.

[0274] Second eluting fraction: (3-methyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester. LC-MS (A): t.sub.R=0.67 min; [M+H].sup.+: 232.16. .sup.1H-NMR (CDCl.sub.3): 8.05 (s, 1H); 7.40-7.30 (m, 5H); 5.23 (s, 0.95H, CH.sub.2); 4.93-4.88 (3s, 2H); 2.42 (s, 3H). Roesy signal seen between CH (triazole) at 8.05 ppm and CH.sub.2 at 4.93-4.88 ppm.

16.2. (3-Methyl-[1, 2, 4]triazol-1-yl)-acetic acid

[0275] This compound was prepared using a method analogous to that of Example 1, step 1.9, (3-methyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester from step 16.1 replacing intermediate 1.8. LC-MS (A): t.sub.R=0.18 min; [M+H].sup.+: 142.22.

16.3. 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methyl-[1, 2, 4]triazol-1-yl)-ethanone

[0276] This compound was prepared using a method analogous to that of Example 11, step 11.3, intermediate 16.2 replacing intermediate 4.2. The crude was purified by Prep LC-MS (11). LC-MS (B): t.sub.R=1.35 min; [M+H].sup.+: 537.1.

Example 17: 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methoxymethyl-[1,2,4]triazol-1-yl)-ethanone

17.1. (3-Methoxymethyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester

[0277] This compound was prepared using a method analogous to that of Example 1, step 1.8, 3-(methoxymethyl)-1H-1,2,4-triazole replacing 3-tert-butyl-1H-1,2,4-triazole. The crude was purified by two CC (1. Biotage, SNAP 10 g cartridge, solvent A: DCM; solvent B: DCM/MeOH 8:2; gradient in % B: 5 for 7CV, 5 to 15 over 3CV, 15 for 3CV. 2. Biotage, SNAP 10 g cartridge, solvent A: DCM; solvent B: DCM/MeOH 8:2; gradient in % B: 5 for 5CV, 5 to 10 over 3CV, 10 for 3CV, 10 to 15 for 3 CV) to yield two regiosiomers:

[0278] First eluting fraction: (5-methoxymethyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester: colourless oil. LC-MS (A): t.sub.R=0.71 min; [M+H].sup.+: 262.2.

[0279] Second eluting fraction: (3-methoxymethyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester: colourless oil. LC-MS (A): t.sub.R=0.67 min; [M+H].sup.+: 262.1. Roesy signal seen between CH (triazole) at 8.17 ppm and NCH.sub.2CO.sub.2 at 5.01 ppm.

17.2. (3-Methoxymethyl-[1, 2, 4]triazol-1-yl)-acetic acid

[0280] This compound was prepared using a method analogous to that of Example 1, step 1.9, (3-methoxymethyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester from step 17.1 replacing intermediate 1.8. LC-MS (A): t.sub.R=0.24 min; [M+H].sup.+: 172.0.

17.3. 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methoxymethyl-[1,2,4]triazol-1-yl)-ethanone

[0281] This compound was prepared using a method analogous to that of Example 11, step 11.3, intermediate 17.2 replacing intermediate 4.2. The crude was purified by Prep LC-MS (II). LC-MS (B): t.sub.R=1.36 min; [M+H].sup.+: 567.1.

Example 18: 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-[3-(5-methyl-[1,2,4]oxadiazol-3-yl)-[1,2,4]triazol-1-yl]-ethanone

18.1. 5-Methyl-3-(1H-[1, 2, 4]triazol-3-yl)-[1, 2, 4]oxadiazole

[0282] To a suspension of N-hydroxy-acetamidine (803 mg) in THF (25 mL) was added NaH (60% in oil, 434 mg) at RT. The foaming suspension was stirred at 70° C. under argon for 30 min, then methyl 1H-1,2,4-triazole-3-carboxylate (725 mg) was added and the mixture was stirred at 70° C. for 2 h40. The reaction mixture was allowed to cool to RT, quenched with sat. NH.sub.4Cl, filtrated off, and the precipitate was washed with THF and MeOH. The filtrate was evaporated and dried at HV to give 1.85 g as beige solid. LC-MS (A): t.sub.R=0.4 min; [M+H].sup.+: 152.2.

18.2. [3-(5-Methyl-[1, 2, 4]oxadiazol-3-yl)-[1,2,4]triazol-1-yl]-acetic acid benzyl ester

[0283] To a suspension of intermediate 18.1 (1.84 g) in MeCN (85 mL) and DMF (5 mL) were added Cs.sub.2CO.sub.3 (3.97 g) and benzyl bromoacetate (4 mL) at RT and the reaction mixture was stirred at RT overnight. The reaction mixture was evaporated to dryness, diluted with EA and washed with water (2×) and brine (1×). The aq. layers were re-extracted with EA (2×). The combined org. layers were dried over MgSO.sub.4, filtrated off, evaporated and dried at HV. CC (Biotage, SNAP 100 g cartridge, solvent A: Hep; solvent B: EA; gradient in % B: 30 for 2CV, 30 to 50 over 3CV, 50 for 3CV, 50 to 70 over 3CV) followed by Prep LC-MS (I) afforded 341 mg as white powder (contains 22 mol % of intermediate 18.3). LC-MS (A): t.sub.R=0.77 min; [M+H].sup.+: 300.0.

18.3. [3-(5-methyl-[1, 2, 4]oxadiazol-3-yl)-[1,2,4]triazol-1-yl]-acetate, as sodium salt

[0284] To a suspension of intermediate 18.2 (335 mg) in EtOH (3 mL) was added 1M NaOH (0.88 mL) at RT and the resulting solution was stirred at RT for 1 h. The reaction mixture was evaporated and dried at HV to give 349 mg as beige solid. LC-MS (A): t.sub.R=0.39 min; [M+H].sup.+: 210.1.

18.4. 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-[3-(5-methyl-[1, 2, 4]oxadiazol-3-yl)-[1, 2, 4]triazol-1-yl]-ethanone

[0285] This compound was prepared using a method analogous to that of Example 11, step 11.3, intermediate 18.3 replacing intermediate 4.2. The crude was purified by Prep LC-MS (I). LC-MS (B): t.sub.R=1.41 min; [M+H].sup.+: 605.1.

Example 19: 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-phenyl-[1,2,4]triazol-1-yl)-ethanone

[0286] This compound was prepared using a method analogous to that of Example 11, step 11.3, 2-(3-phenyl-1H-1,2,4-triazol-1-yl)acetic acid replacing intermediate 4.2. The crude was purified by Prep LC-MS (I). LC-MS (C): t.sub.R=1.15 min; [M+H].sup.+: 599.2.

Example 20: 2-(3-Acetyl-[1,2,4]triazol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

20.1. (3-Acetyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester

[0287] This compound was prepared using a method analogous to that of Example 1, step 1.8, 1-(1H-1,2,4-triazol-5-yl)ethanone replacing 3-tert-butyl-1H-1,2,4-triazole. The crude was purified by CC (Biotage, SNAP 10 g cartridge, solvent A: Hep; solvent B: EA; gradient in % B: 30 for 4CV, 30 to 70 over 4CV, 70 for 2CV, 70 to 100 over 2CV, 100 for 2CV) to give the desired triazole regioisomer as second fraction. Roesy signal seen between CH (triazole) at 8.28 ppm and CH.sub.2 at 5.1 ppm.

[0288] LC-MS (A): t.sub.R=0.7 min; [M+H].sup.+: 260.1.

20.2. (3-Acetyl-[1,2, 4]triazol-1-yl)-acetic acid

[0289] This compound was prepared using a method analogous to that of Example 1, step 1.9, intermediate 20.1 replacing intermediate 1.8. LC-MS (A): t.sub.R=0.25 min; [M+H].sup.+: 170.0.

20.3. 2-(3-Acetyl-[1, 2, 4]triazol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0290] This compound was prepared using a method analogous to that of Example 11, step 11.3, intermediate 20.2 replacing intermediate 4.2 and DCM replacing DMF. The crude was purified by Prep LC-MS (11) followed by Prep LC-MS (IV). LC-MS (B): t.sub.R=1.38 min; [M+H].sup.+: 565.1.

Example 21: 2-[3-(1-Hydroxy-ethyl)-[1,2,4]triazol-1-yl]-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0291] To solution of Example 20 (40 mg) in THF (0.75 mL) and EtOH (0.25 mL) was added NaBH.sub.4 (1.3 mg) and the mixture was stirred at 0° C. for 1 h10. The reaction mixture was quenched by addition of H.sub.2O and extracted with DCM (3×). The combined org. layers were evaporated and dried at HV. The crude was purified by Prep LC-MS (IV) to give 25 mg as white powder. LC-MS (B): t.sub.R=1.32 min; [M+H].sup.+: 567.1.

Example 22: 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methyl-pyrazol-1-yl)-ethanone

[0292] This compound was prepared using a method analogous to that of Example 11, step 11.3, (3-methyl-1H-pyrazol-1-yl)acetic acid replacing intermediate 4.2 and DCM replacing DMF. The crude was purified by Prep LC-MS (IV). LC-MS (C): t.sub.R=1.07 min; [M+H].sup.+: 536.4.

Example 23: 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-pyridin-2-yl-[1,2,4]triazol-1-yl)-ethanone

23.1. (3-Pyridin-2-yl-[1, 2, 4]triazol-1-yl)-acetic acid tert-butyl ester

[0293] This compound was prepared using a method analogous to that of Example 1, step 1.8, 2-(1H-1,2,4-triazol-3-yl)pyridine replacing 3-tert-butyl-1H-1,2,4-triazole and tert-butyl bromoacetate replacing benzyl bromoacetate. The compound was further purified by Prep LC-MS (VI). LC-MS (A): t.sub.R=0.61 min; [M+H].sup.+: 261.2.

23.2. 2-(3-(Pyridin-2-yl)-H-1,2,4-triazol-1-yl)acetic acid, as hydrochloride salt

[0294] To a suspension of intermediate 23.1 (263 mg) dioxane (2 mL) was added HCl (4M in dioxane; 2 mL). After 1 h at RT, few drops of H.sub.2O were added and the reaction mixture was stirred for 26 h. The reaction mixture was evaporated and dried at HV to give 263 mg as yellowish solid. LC-MS (A): t.sub.R=0.29 min; [M+H].sup.+: 205.1.

23.3. 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-pyridin-2-yl-[1, 2, 4]triazol-1-yl)-ethanone

[0295] This compound was prepared using a method analogous to that of Example 11, step 11.3, intermediate 23.2 replacing intermediate 4.2 and DCM replacing DMF. The crude was purified by Prep LC-MS (IV) followed by Prep TLC (0.5 mm, DCM/MeOH 95/5). LC-MS (C): t.sub.R=1.00 min; [M+H].sup.+: 600.1.

Example 24 and Example 25: 2-(3-Ethyl-5-methyl-[1,2,4]triazol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone and 2-(5-Ethyl-3-methyl-[1,2,4]triazol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0296] Acid Precursors for Example 24 and 25:

[0297] Step 24.1. To ethylacetimidate hydrochloride (500 mg) in MeCN (6 mL) was added Amberlyst A21 (1.14 g) and the suspension stirred at RT for 15 min. Then the mixture was filtrated and the resin washed with MeCN (1 mL). Propanoic acid hydrazide (353 mg) was added to the filtrate and the resulting white suspension was stirred at 50° C. under argon for 4 d and at 80° C. overnight. The reaction mixture was evaporated. Purification by CC (Biotage, SNAP 10 g cartridge, solvent A: DCM; solvent B: MeOH; gradient in % B: 10 for 6CV, 10 to 20 over 3CV, 20 for 6CV) followed by a second CC (Biotage, SNAP 10 g cartridge, solvent A: DCM; solvent B: DCM/MeOH 8:2; gradient in % B: 25 for 7CV, 25 to 50 over 3CV, 50 for 5CV) afforded 3-ethyl-5-methyl-1H-[1,2,4]triazole (125 mg as yellow oil). LC-MS (A): t.sub.R=0.21 min; [M+H].sup.+: 112.4.

[0298] Step 24.2: A method analogous to that of Example 1 step 1.8 was followed, 3-ethyl-5-methyl-1H-[1,2,4]triazole from step 24.1 replacing 3-tert-butyl-1H-1,2,4-triazole. A mixture of regioisomers was obtained: (3-ethyl-5-methyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester and (5-ethyl-3-methyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester.

[0299] Step 24.3: A method analogous to that of Example 1 step 1.9 was followed, intermediates from step 24.2 replacing intermediate 1.8. A mixture of regioisomers (ca. 1:1) was obtained: (3-ethyl-5-methyl-[1,2,4]triazol-1-yl)-acetic acid and (5-ethyl-3-methyl-[1,2,4]triazol-1-yl)-acetic acid.

[0300] The final compounds were prepared using a method analogous to that of Example 11, step 11.3, intermediates from step 24.3 replacing intermediate 4.2 and DCM replacing DMF. The crude was purified by Prep LC-MS (IV) followed by preparative chiral HPLC (I). Both fractions were taken up in DCM and washed with water (2×) and the aq. layers extracted with DCM (1×). The combined org. layers were dried over MgSO.sub.4, filtrated off, evaporated and dried at HV.

[0301] First eluting fraction (Example 24): 2-(3-ethyl-5-methyl-[1,2,4]triazol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone. LC-MS (C): t.sub.R=1.00 min; [M+H].sup.+: 565.1.

[0302] Second eluting fraction (Example 25): 2-(5-ethyl-3-methyl-[1,2,4]triazol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone. LC-MS (C): t.sub.R=1.00 min; [M+H].sup.+: 565.5.

Example 26: 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-imidazo[4,5-b]pyridin-3-yl-ethanone

26.1. Imidazo[4, 5-b]pyridin-3-yl-acetic acid benzyl ester

[0303] To a solution of 4-azabenzimidazole (4.75 g) in DMF (80 mL) was added benzyl bromoacetate (6.58 mL) followed by Cs.sub.2CO.sub.3 (25.9 g). The resulting suspension was stirred overnight. The reaction mixture was diluted with EA and washed with water (2×) and aq. sat. NH.sub.4Cl. The aq. layers were extracted with EA (2×). The combined org. layers were dried over MgSO.sub.4, filtrated off and evaporated in reduced pressure. The crude was purified by CC (Biotage, SNAP 100 g cartridge, solvent A: DCM; solvent B: DCM/MeOH 8:2; gradient in % B: 0 to 5 over 3CV, 5 for 5CV, 5 to 15 over 5CV, 15 for 3CV) to afford 4.99 g of the desired compound as yellow solid. LC-MS (B): t.sub.R=0.59 min; [M+H].sup.+: 267.86.

26.2. Imidazo[4, 5-b]pyridin-3-yl-acetic acid

[0304] To a yellow suspension of intermediate 26.1 (4.99 g) in MeOH (30 mL) and acetic acid (0.3 mL) was added Pd/C (10%, 994 mg) under argon. The flask was evacuated and backfilled with argon three times, then evacuated and backfilled with hydrogen twice. The reaction mixture was stirred at RT under hydrogen for 5 h, filtrated over celite and the celite was washed with MeOH. The filtrate was evaporated to dryness to afford 2.41 g of off-white solid that was used without purification. LC-MS (B): t.sub.R=0.15 min; [M+H].sup.+: 178.24.

26.3. 1-{(R)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-imidazo[4, 5-b]pyridin-3-yl-ethanone

[0305] This compound was prepared using a method analogous to that of Example 11, step 11.3, intermediate 26.2 replacing intermediate 4.2 and DCM replacing DMF. The crude was purified by Prep LC-MS (IV) followed by Prep TLC (0.5 mm, DCM/MeOH 95/5). LC-MS (A): t.sub.R=0.8 min; [M+H].sup.+: 573.0.

Example 27: 2-(3,5-Dimethyl-[1,2,4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

27.1. (R)-2-(4-(2-(trifluoromethyl)-4-(2-(trifluoromethyl)pyrimidin-5-yl)thiazol-5-yl)piperazin-2-yl)ethan-1-ol, as hydrochloride salt

[0306] This compound was prepared using a method analogous to that of Example 1, following steps 1.1 to steps 1.7, (R)-1-Boc-2-hydroxyethylpiperazine replacing (S)-1-Boc-2-hydroxymethylpiperazine in step 1.3. LC-MS (A): t.sub.R=0.69 min; [M+H].sup.+: 428.0.

27.2. 2-(3, 5-Dimethyl-[1, 2, 4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0307] This compound was prepared using a method analogous to that of Example 12, intermediate 27.1 replacing intermediate 11.2 and DCM replacing DMF. The crude was purified by Prep LC-MS (I) followed by Prep TLC (0.5 mm, DCM/MeOH 97/3). LC-MS (C): t.sub.R=0.98 min; [M+H].sup.+: 565.2.

Example 28: 1-{(R)-2-(2-Hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-trifluoromethyl-[1,2,4]triazol-1-yl)-ethanone

[0308] This compound was prepared using a method analogous to that of Example 27, step 27.2, intermediate 5.2 replacing (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid. The crude was purified by Prep LC-MS (V). LC-MS (C): t.sub.R=1.16 min; [M+H].sup.+: 605.1.

Example 29: 2-(3-Cyclobutyl-[1,2,4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

29.1. (3-Cyclobutyl-[1, 2, 4]triazol-1-yl)-acetic acid benzyl ester

[0309] This compound was prepared using a method analogous to that of Example 1, step 1.8, 3-cyclobutyl-1H-1,2,4-triazole replacing of 3-tert-butyl-1H-1,2,4-triazole. A further Prep LC-MS (IV) purification was performed before submitting the regioisomeric mixture for separation to preparative chiral HPLC (V).

[0310] First eluting fraction: (5-cyclobutyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester. LC-MS (A): t.sub.R=0.68 min; [M+H].sup.+: 272.2.

[0311] Second eluting fraction: (3-cyclobutyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester. (LC-MS (A): t.sub.R=0.7 min; [M+H].sup.+: 272.2.

29.2. (3-Cyclobutyl-[1, 2, 4]triazol-1-yl)-acetic acid

[0312] This compound was prepared using a method analogous to that of Example 1, step 1.9, (3-cyclobutyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester from step 29.1 replacing intermediate 1.8. LC-MS (A): t.sub.R=0.32 min; [M+H].sup.+: 182.3. Roesy signal seen between CH (triazole) at 8.40 ppm and N-CH2-COOH at 5.02 ppm.

29.3. 2-(3-Cyclobutyl-[1, 2, 4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0313] This compound was prepared using a method analogous to that of Example 27, step 27.2, intermediate 29.2 replacing (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid. The crude was purified by Prep LC-MS (IV). LC-MS (C): t.sub.R=1.11 min; [M+H].sup.+: 591.2.

Example 30: 2-(3-tert-Butyl-[1,2,4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0314] This compound was prepared using a method analogous to that of Example 27, step 27.2, intermediate 1.9 replacing (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid. The crude was purified by Prep LC-MS (I). LC-MS (C): t.sub.R=1.13 min; [M+H].sup.+: 593.2.

Example 31: 2-(3-Cyclopropyl-[1,2,4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0315] This compound was prepared using a method analogous to that of Example 27, step 27.2, intermediate 3.2 replacing (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid. The crude was purified by Prep LC-MS (I). LC-MS (C): t.sub.R=1.06 min; [M+H].sup.+: 577.4.

Example 32: 2-(3-Ethyl-[1,2,4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

32.1. (3-Ethyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester

[0316] This compound was prepared using a method analogous to that of Example 1, step 1.8, 3-ethyl-1H-1,2,4-triazole replacing of 3-tert-butyl-1H-1,2,4-triazole. The mixture of regioisomers was purified by preparative chiral HPLC (VI). First eluting fraction: (5-ethyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester: LC-MS (A): t.sub.R=0.72 min; [M+H].sup.+: 246.2. Roesy signal seen between CH.sub.2CH.sub.3 at 2.72 ppm and CH.sub.2CO.sub.2 at 4.93 ppm.

[0317] Second eluting fraction: (3-ethyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester: LC-MS (A): t.sub.R=0.71 min; [M+H].sup.+: 246.2. Roesy signal seen between CH at 8.08 ppm and CH.sub.2CO.sub.2 at 4.96 ppm.

32.2. (3-Ethyl-[1, 2, 4]triazol-1-yl)-acetic acid

[0318] This compound was prepared using a method analogous to that of Example 1, step 1.9, (3-ethyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester from step 32.1 replacing intermediate 1.8. LC-MS (A): t.sub.R=0.25 min; [M+H].sup.+: 156.2.

32.3. 2-(3-Ethyl-[1, 2, 4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0319] This compound was prepared using a method analogous to that of Example 27, step 27.2, intermediate 32.2 replacing (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid. The crude was purified by Prep LC-MS (IV). LC-MS (C): t.sub.R=1.04 min; [M+H].sup.+: 564.9.

Example 33: 2-(5-Ethyl-[1,2,4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

33.1. (5-Ethyl-[1, 2, 4]triazol-1-yl)-acetic acid

[0320] This compound was prepared using a method analogous to that of Example 1, step 1.9, (5-ethyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester from step 32.1 replacing intermediate 1.8. LC-MS (A): t.sub.R=0.22 min; [M+H].sup.+: 156.1.

33.2. 2-(5-Ethyl-[1,2,4]triazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0321] This compound was prepared using a method analogous to that of Example 27, step 27.2, intermediate 33.1 replacing (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid. The crude was purified by Prep LC-MS (I). LC-MS (C): t.sub.R=1.03 min; [M+H].sup.+: 565.2.

Example 34: 1-{(R)-2-(2-Hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-isopropyl-[1,2,4]triazol-1-yl)-ethanone

[0322] This compound was prepared using a method analogous to that of Example 27, step 27.2, intermediate 4.2 replacing (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid. The crude was purified by Prep LC-MS (IV). LC-MS (C): t.sub.R=1.08 min; [M+H].sup.+: 579.2.

Example 35: 1-{(R)-2-(2-Hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methyl-[1,2,4]triazol-1-yl)-ethanone

[0323] This compound was prepared using a method analogous to that of Example 27, step 27.2, intermediate 16.2 replacing (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid. The crude was purified by Prep LC-MS (11) followed by Prep LC-MS (IV). LC-MS (C): t.sub.R=1.00 min; [M+H].sup.+: 551.2.

Example 36: 1-{(R)-2-(2-Hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(5-methyl-[1,2,4]triazol-1-yl)-ethanone

36.1. (5-Methyl-[1, 2, 4]triazol-1-yl)-acetic acid

[0324] This compound was prepared using a method analogous to that of Example 1, step 1.9, (5-methyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester from step 16.1 replacing intermediate 1.8. LC-MS (A): t.sub.R=0.19 min; [M+H].sup.+: 142.2.

36.2. 1-{(R)-2-(2-Hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(5-methyl-[1, 2, 4]triazol-1-yl)-ethanone

[0325] This compound was prepared using a method analogous to that of Example 27, step 27.2, intermediate 36.1 replacing (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid. The crude was purified by Prep LC-MS (IV). LC-MS (C): t.sub.R=1.00 min; [M+H].sup.+: 551.1.

Example 37: 1-{(R)-2-(2-Hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methoxymethyl-[1,2,4]triazol-1-yl)-ethanone

[0326] This compound was prepared using a method analogous to that of Example 27, step 27.2, intermediate 17.2 replacing (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid. The crude was purified by Prep LC-MS (IV). LC-MS (B): t.sub.R=1.37 min; [M+H].sup.+: 581.1.

Example 38: 2-(3-Difluoromethyl-[1,2,4]triazol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

38.1. (3-Difluoromethyl-[1, 2,4]triazol-1-yl)-acetic acid benzyl ester

[0327] This compound was prepared using a method analogous to that of Example 1, step 1.8, 3-(difluoromethyl)-1H-1,2,4-triazole replacing 3-tert-butyl-1H-1,2,4-triazole. The crude was purified by CC (Biotage, SNAP 50 g cartridge, solvent A: Hep; solvent B: EA; gradient in % B: 30 for 5CV, 30 to 70 over 5CV, 70 for 3CV, 70 to 100 over 2CV, 100 for 1CV) to yield two regiosiomers:

[0328] First eluting fraction: (5-difluoromethyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester: yellow oil. LC-MS (A): t.sub.R=0.78 min; [M+H].sup.+: 268.2.

[0329] Second eluting fraction: (3-difluoromethyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester: off-white solid. LC-MS (A): t.sub.R=0.77 min; [M+H].sup.+: 268.1.

38.2. (3-Difluoromethyl-[1, 2,4]triazol-1-yl)-acetic acid

[0330] This compound was prepared using a method analogous to that of Example 1, step 1.9, (3-difluoromethyl-[1,2,4]triazol-1-yl)-acetic acid benzyl ester from step 38.1 replacing intermediate 1.8. LC-MS (A): t.sub.R=0.26 min; [M+H].sup.+: 178.2.

38.3. 2-(3-Difluoromethyl-[1, 2, 4]triazol-1-yl)-1-{(R)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0331] This compound was prepared using a method analogous to that of Example 27, step 27.2, intermediate 38.2 replacing (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid. The crude was purified by Prep LC-MS (IV). LC-MS (A): t.sub.R=0.86 min; [M+H].sup.+: 573.0.

Example 39: 1-{(R)-2-(2-Hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methyl-pyrazol-1-yl)-ethanone

[0332] This compound was prepared using a method analogous to that of Example 27, step 27.2, (3-methyl-1H-pyrazol-1-yl)acetic acid replacing (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid. The crude was purified by Prep LC-MS (IV). LC-MS (C): t.sub.R=1.08 min; [M+H].sup.+: 550.0.

Example 40: 2-(3-Ethyl-pyrazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

40.1 (3-Ethyl-pyrazol-1-yl)-acetic acid benzyl ester

[0333] This compound was prepared using a method analogous to that of Example 1, step 1.8, 3-ethyl-1H-pyrazole replacing 3-tert-butyl-1H-1,2,4-triazole. The mixture of regioisomers was purified by preparative chiral HPLC (VII). First eluting fraction: (3-ethyl-pyrazol-1-yl)-acetic acid benzyl ester: LC-MS (A): t.sub.R=0.82 min; [M+H].sup.+: 245.1. Roesy signal seen between pyrazol-CH at 7.37 ppm and CH.sub.2CO.sub.2 at 4.91 ppm.

40.2 (3-Ethyl-pyrazol-1-yl)-acetic acid

[0334] This compound was prepared using a method analogous to that of Example 1, step 1.9, (3-ethyl-pyrazol-1-yl)-acetic acid benzyl ester from step 40.1 replacing intermediate 1.8. LC-MS (A): t.sub.R=0.43 min; [M+H].sup.+: 155.4.

40.3. 2-(3-Ethyl-pyrazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0335] This compound was prepared using a method analogous to that of Example 27, step 27.2, intermediate 40.2 replacing (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid. The crude was purified by Prep LC-MS (I). LC-MS (C): t.sub.R=1.14 min; [M+H].sup.+: 564.0.

Example 41: 2-(3-Cyclopropyl-pyrazol-1-yl)-1-{(R)-2-(2-hydroxy-ethyl)-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0336] This compound was prepared using a method analogous to that of Example 27, step 27.2, (3-cyclopropyl-1H-pyrazol-1-yl)acetic acid replacing (3,5-dimethyl-1H-1,2,4-triazol-1-yl)acetic acid. The crude was purified by Prep LC-MS (IV). LC-MS (C): t.sub.R=1.15 min; [M+H].sup.+: 576.2.

Example 42: 1-{(S)-2-Hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-2-(3-methyl-pyrazol-1-yl)-ethanone

[0337] This compound was prepared using a method analogous to that of Example 1, step 1.10, (3-methyl-1H-pyrazol-1-yl)acetic acid replacing intermediate 1.9. The crude was purified by Prep LC-MS (I). LC-MS (C): t.sub.R=1.07 min; [M+H].sup.+: 536.2.

Example 43: 2-(3-Ethyl-pyrazol-1-yl)-1-{(S)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0338] This compound was prepared using a method analogous to that of Example 1, step 1.10, intermediate 40.2 replacing intermediate 1.9. The crude was purified by Prep LC-MS (I). LC-MS (C): t.sub.R=1.12 min; [M+H].sup.+: 550.0.

Example 44: 2-(3-Cyclopropyl-pyrazol-1-yl)-1-{(S)-2-hydroxymethyl-4-[2-trifluoromethyl-4-(2-trifluoromethyl-pyrimidin-5-yl)-thiazol-5-yl]-piperazin-1-yl}-ethanone

[0339] This compound was prepared using a method analogous to that of Example 1, step 1.10, (3-cyclopropyl-1H-pyrazol-1-yl)acetic acid replacing intermediate 1.9. The crude was purified by Prep LC-MS (I). LC-MS (C): t.sub.R=1.13 min; [M+H].sup.+: 562.2.

[0340] II. Biological Assays

[0341] A) FLIPR assay: The bioactivity of compounds is tested in a fluorometric imaging plate reader (FLIPR: Molecular Devices) using engineered CHO-K1 cells expressing the human CXCR3A (GenBank: AY242128) coupled to a G protein (Galpha(16)). Cells are plated the day prior to bioassay in F12 medium supplemented with 10% FBS and G418 and hygromycin antibiotics to maintain recombinant selection. At the day of bioassay, cells are washed and dye loaded for one hour with Fluo-4-AM (Invitrogen) in Hanks Balanced Salt Solution (Invitrogen), buffered with 20 mM Hepes at pH 7.4 and sodium bicarbonate (0.038%), containing 5 mM probenecid. This buffer, but lacking the dye and containing probenecid at a concentration of 2.5 mM, is also used for washing steps (wash buffer); or lacking both dye and probenecid but supplemented with 0.1% BSA for compound dilution steps (dilution buffer). Cells are washed free of excess dye and 60 microliter of wash buffer is added. Stock solutions of test compounds are made up at a concentration of 10 mM in DMSO, and serially diluted in dilution buffer to concentrations required for inhibition dose response curves. After a 10 minute incubation period at 37° C., 10 microliters of each compound dilution are transferred from a compound plate to the plate containing the recombinant cells in the FLIPR instrument according to the manufacturer's instructions. Following basal readings, 10 microliter CXCL10 agonist at a concentration of 20 nM (from Peprotech) is added, again using the FLIPR instrument. Changes in fluorescence are monitored before and after addition of the test compounds. Emission peak values above base level after CXCL10 addition are exported after base line subtraction.

[0342] The calculated IC.sub.50 values may fluctuate depending on the daily assay performance. Fluctuations of this kind are known to those skilled in the art. In the case where IC.sub.50 values have been determined several times for the same compound, mean values are given. Data are shown in Table 1.

TABLE-US-00010 TABLE 1 Example No FLIPR: IC.sub.50 (nM) 1 6.7 2 6.0 3 3.0 4 1.8 5 4.0 6 4.6 7 3.1 8 10 9 1.3 10 1.1 11 3.4 12 6.9 13 1.2 14 0.7 15 2.3 16 3.9 17 5.9 18 510 19 42 20 6.4 21 4.3 22 1.6 23 35 24 7.6 25 3.6 26 3.5 27 5.2 28 1.1 29 5.5 30 4.4 31 4.4 32 2.6 33 27 34 4.5 35 2.9 36 8.0 37 5.7 38 8.6 39 1.0 40 1.6 41 0.45 42 1.0 43 1.2 44 0.38

[0343] B): Receptor internalization assay: Stock solutions of test compounds are made up at a concentration of 10 mM in DMSO, and serially diluted in PBS containing 0.5% BSA to concentrations required for inhibition dose response curves. Diluted compounds are then mixed with an equal volume of CXCL10 (Peprotech) diluted in PBS. Anticoagulated venous human whole blood is added to the mixture, which is then incubated in a CO.sub.2 incubator at 37° C. to allow for ligand mediated receptor internalization (final CXCL10 concentration is 9 nM). After 30 min, the blood is mixed with fluorescently labeled CXCR3 and CD4 specific antibodies (Becton Dickinson) and incubated on ice for 10 minutes. Samples are then mixed with BD FACS Lysing Solution (Becton Dickinson) in order to eliminate red blood cells. After washing the cells with PBS containing 0.5% BSA, the samples are then analyzed in a flow cytometer (FACS Canto II, Becton Dickinson). For data analysis using FACSDiva software (Becton Dickinson), the mean fluorescence corresponding to CXCR3 cell surface expression was determined on CD4 positive cells. The program GraphPad Prism or similar software is used to fit the data to a single site dose response curve and to calculate IC.sub.50 values.

[0344] The calculated IC.sub.50 values may fluctuate depending on the daily assay performance. Fluctuations of this kind are known to those skilled in the art. In the case where IC.sub.50 values have been determined several times for the same compound, mean values are given. Data are shown in Table 2.

TABLE-US-00011 TABLE 2 Example No Internalization IC.sub.50 (nM) 1 415 2 463 3 191 4 283 5 328 6 778 7 2270 8 3200 9 1520 10 961 11 339 12 324 13 1050 14 199 15 1660 16 542 17 1290 18 6500 19 3310 20 2380 21 737 22 105 23 2730 24 355 25 664 26 74.2 27 235 28 214 29 157 30 170 31 73.8 32 93.3 33 652 34 84.8 35 102 36 451 37 393 38 258 39 46.3 40 32.7 41 30.2 42 59.7 43 40.3 44 76.0

[0345] C) hERG Q-Patch assay: Compounds are evaluated for block of the hERG K channel using CHO cells stably expressing the hERG gene (accession number U04270, bSys, Witterswil, Switzerland) and the QPatch robotic platform (Sophion, Ballerup, Denmark) in single-cell mode at room temperature. Cells are grown in culture flasks at 37° C. in 5% CO.sub.2, in culture medium (Ham's F-12 Nutrient Mixture, Invitrogen 21765-029) supplemented with 9% (v/v) fetal calf serum, 0.9% Penicillin/Streptomycin (10,000 U/mL, Invitrogen 15140148), 100 μg/mL Hygromycin B (Invitrogen 10687010). When the cells are 80% confluent (every 2-3 days), they are either split for further culture or used for electrophysiology. For further culture, cells are detached with 0.25% Trypsin EDTA solution (Invitrogen 25200-056) and a fraction of the cells (10-30%) is reseeded in culture medium. For electrophysiology, on the experimental day, cells are detached with 0.25% Trypsin EDTA solution and all cells are suspended in suspension medium (293 SFM II, Invitrogen 11686-029) supplemented with 20 mM HEPES and 0.04 mg/mL Trypsin inhibitor. Cells are kept in suspension medium at 32-35° C. in the QPatch robot until use, at which time aliquots are transferred to the extracellular solution (in mM: NaCl 150; KCl 4; CaCl.sub.2 1.2; MgCl.sub.2 1; HEPES 10; pH 7.4 with NaOH) containing 0.3% v/v DMSO and applied to the test plates. K+ currents are measured with the patch-voltage-clamp technique in the whole-cell configuration with the internal solution (in mM: KCl, 140; NaCl, 10; MgCl.sub.2, 1; HEPES, 10: EGTA, 5; pH=7.2 with KOH). Currents are low-pass filtered using the internal Bessel filter of the QPatch robot with a cut-off frequency of 2 kHz and are digitized at 10 kHz. K.sup.+ tail currents are produced from a holding voltage of −80 mV by a 500-ms depolarization to +20 mV followed by a 500-ms repolarization to −40 mV; tail current amplitudes are measured at the end of the repolarization to −40 mV. The pulse pattern is repeated every 10 sec during the experiment, baseline K.sup.+ current is measured after 3 min in extracellular solution, test-solution containing compound is then applied, and K.sup.+ current in presence of compound is measured 3 minutes after application to the cells. The respective test-solution is prepared by (1) dissolving the test-compound in pure DMSO, (2) diluting this DMSO solution in extracellular solution, and (3) adding further DMSO, such that the final test-solution has a concentration of either 300 nM or 3000 nM of the test-compound and contains 0.3% v/v DMSO. Compound effects are quantified as % block by dividing the current in presence of compound by the baseline current; two or three experiments are performed for each compound and the final value represents the mean of the results of each experiment. Data are shown in Table 3.

TABLE-US-00012 TABLE 3 concentration concentration Example No [nM] % block [nM] % block 1 300 2 3000 12 2 300 −3 3000 −4 3 300 0 3000 12 4 300 4 3000 13 5 300 −2 3000 7 6 300 13 3000 23 7 300 24 3000 45 8 300 19 3000 54 9 300 8 3000 24 10 300 2 3000 12 11 300 5 3000 11 12 300 1 3000 3 13 300 5 3000 17 14 300 14 3000 62 15 300 8 3000 35 16 300 1 3000 1 17 300 2 3000 6 18 300 6 3000 13 19 300 6 3000 41 20 300 2 3000 3 22 300 4 3000 15 27 300 1 3000 4 28 300 6 3000 40 29 300 4 3000 38 30 300 −2 3000 25 31 300 2 3000 23 32 300 0 3000 19 33 300 −5 3000 6 34 300 10 3000 34 35 300 11 3000 25 36 300 10 3000 19 37 300 4 3000 9 39 300 1 3000 32 40 300 5 3000 47 41 300 12 3000 60 42 300 2 3000 15 43 300 5 3000 25 44 300 5 3000 34