Hydroxyalkyl-substituted phenyltriazole derivatives and uses thereof

Abstract

The present invention relates to novel 5-(hydroxyalkyl)-1-phenyl-1,2,4-triazole derivatives, to processes for the preparation of such compounds, to pharmaceutical compositions containing such compounds, and to the use of such compounds or compositions for the treatment and/or prevention of diseases, in particular for the treatment and/or prevention of cardiovascular and renal diseases.

Claims

1. A compound of formula (I) ##STR00061## wherein R.sup.1is hydrogen or methyl, and R.sup.2A and R.sup.2B are independently selected from the group consisting of hydrogen, fluoro, chloro, cyano, methyl, fluoromethyl, difluoromethyl, trifluoromethyl, ethyl, methoxy, difluoromethoxy and trifluoromethoxy, or a pharmaceutically acceptable salt, hydrate and/or solvate thereof.

2. The compound of claim 1, wherein R.sup.1 is hydrogen or methyl, and R.sup.2A and R.sup.2B are independently selected from the group consisting of hydrogen, fluoro, chloro, methyl and methoxy, wherein at least one of R.sup.2A and R.sup.2B is other than hydrogen, or a pharmaceutically acceptable salt, hydrate and/or solvate thereof.

3. The compound of claim 1, wherein the compound is selected from the group consisting of 5-(4-chlorophenyl)-2-{[1-(3-chlorophenyl)-5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]-methyl-}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 5-(4-chlorophenyl)-2-{[1-(3-fluorophenyl)-5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]-methyl-}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 5 -(4-chlorophenyl)-2-{[5 -(hydroxymethyl)-1-(2-methylphenyl)-1H-1,2,4-triazol-3-yl]-methyl-}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 2-({1-(2-chloro-4-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-5 -(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 2-{[1-(2-chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one(diastereomer 1); 2-{[1-(2-chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one(diastereomer 2); 2-({1-(2-chloro-5-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 2-{[1-(2-chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one(diastereomer 1); 2-{[1-(2-chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one(diastereomer 2); 5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1R)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1R)- 1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1R)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; and 5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; or a pharmaceutically acceptable salt, hydrate and/or solvate thereof.

4. The compound of claim 1, wherein the compound is selected from the group consisting of 5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; 5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; and 5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one; or a pharmaceutically acceptable salt, hydrate and/or solvate thereof.

5. 5-(4-Chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one, a compound having the following formula ##STR00062## or a pharmaceutically acceptable salt, hydrate and/or solvate thereof.

6. The compound of claim 5, wherein the compound is 5-(4-Chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one having the following formula ##STR00063##

7. A process for preparing a compound of claim 1, comprising: reacting a compound of formula (II) ##STR00064## is with hydrazine to give the hydrazide of formula (III) ##STR00065## condensing the hydrazide of formula (III) with an amidine of formula (IV) ##STR00066## or a salt thereof, wherein R.sup.1 has the meaning indicated in claim 1, in the presence of a base to give a 1,2,4-triazole derivative of formula (V) ##STR00067## and/or a tautomer thereof, wherein R.sup.1 has the meaning indicated in claim 1, and coupling the 1,2,4-triazole derivative of formula (V) and/or tautomer thereof with a phenylboronic acid of formula (VI) ##STR00068## wherein R.sup.2A and R.sup.2B have the meanings indicated in claim 1, in the presence of a copper catalyst and an amine base to yield a compound of formula (I) ##STR00069## wherein R.sup.1, R.sup.2Aand R.sup.2B have the meanings indicated in claim 1, and when more than one diastereomer, hydrate, solvate, salt, or hydrate or solvate of the salt of the compound of formula (I) is produced they are optionally separated into their respective diastereomers, and/or converted into their respective hydrates, solvates, salts and/or hydrates or solvates of the salts by treatment with the corresponding solvents and/or acids or bases.

8. A pharmaceutical composition comprising a compound of claim 1 and at least one pharmaceutically acceptable excipient.

9. The pharmaceutical composition of claim 8 further comprising at least one therapeutic agent selected from the group consisting of a diuretic, an angiotensin AII antagonist, an ACE inhibitor, a beta-receptor blocker, a mineralocorticoid receptor antagonist, an organic nitrate, an NO donor, an activator of soluble guanylate cyclase, a stimulator of soluble guanylate cyclase, and a positive-inotropic agent.

10. A method of inhibiting the action of vasopressin in a human or other animal comprising administering to the human or the other mammal in need thereof a therapeutically effective amount of at least one compound of claim 1.

11. A method of inhibiting the action of vasopressin in a human or other animal comprising administering to the human or the other mammal in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 8.

12. A method for the treatment of acute and chronic heart failure, comprising administering to a human or other mammal in need thereof a therapeutically effective amount of at least one compound of claim 1.

13. A method for the treatment of acute and chronic heart failure, comprising administering to a human or other mammal in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 8.

14. A method for treatment of acute and chronic heart failure, comprising administering to a human or other mammal in need thereof a therapeutically effective amount of 5-(4-Chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one, a compound having the following formula ##STR00070## or a pharmaceutically acceptable salt, hydrate and/or solvate thereof.

15. The method of claim 14, comprising administering the compound, having the following formula ##STR00071##

Description

A. EXAMPLES

(1) Abbreviations and Acronyms: Ac acetyl aq. aqueous (solution) br. broad (.sup.1H NMR signal) cat. catalytic conc. concentrated d doublet (.sup.1H NMR signal) DCI direct chemical ionization (MS) d.e. diastereomeric excess DMF N,N-dimethylformamide DMSO dimethylsulfoxide EI electron impact ionization (MS) eq. equivalent(s) ESI electro-spray ionization (MS) Et ethyl h hour(s) .sup.1H NMR proton nuclear magnetic resonance spectroscopy HPLC high performance liquid chromatography LC/MS liquid chromatography-coupled mass spectroscopy m multiplet (.sup.1H NMR signal) Me methyl min minute(s) MS mass spectroscopy MTBE methyl tert-butyl ether m/z mass-to-charge ratio (MS) of th. of theory (chemical yield) q quartet (.sup.1H NMR signal) quant. quantitative (yield) rac racemic Rf TLC retention factor RP reverse phase (HPLC) rt room temperature R.sub.t retention time (HPLC) s singlet (.sup.1H NMR signal) sat. saturated (solution) SFC supercritical fluid chromatography t triplet (.sup.1H NMR signal) tBu tert-butyl tert tertiary TFA trifluoroacetic acid THF tetrahydrofuran TLC thin layer chromatography UV ultraviolet

(2) LC/MS and HPLC Methods:

(3) Method 1 (LC/MS):

(4) Instrument: Waters Acquity SQD UPLC System; column: Waters Acquity UPLC HSS T3 1.8μ, 50 mm×1 mm; eluent A: 1 L water+0.25 mL 99% formic acid, eluent B: 1 L acetonitrile+0.25 mL 99% formic acid; gradient: 0.0 min 90% A.fwdarw.1.2 min 5% A.fwdarw.2.0 min 5% A; oven: 50° C.; flow rate: 0.40 mL/min; UV detection: 208-400 nm.

(5) Method 2 (LC/MS):

(6) Instrument: Waters Acquity SQD UPLC System; column: Waters Acquity UPLC HSS T3 1.8μ, 50 mm×1 mm; eluent A: 1 L water+0.25 mL 99% formic acid, eluent B: 1 L acetonitrile+0.25 mL 99% formic acid; gradient: 0.0 min 95% A.fwdarw.6.0 min 5% A.fwdarw.7.5 min 5% A; oven: 50° C.; flow rate: 0.35 mL/min; UV detection: 210-400 nm.

(7) Method 3 (LC/MS):

(8) Instrument MS: Agilent MS Quad 6150; Instrument HPLC: Agilent 1290; column: Waters Acquity UPLC HSS T3 1.8μ, 50 mm×2.1 mm; eluent A: 1 L water+0.25 mL 99% formic acid, eluent B: 1 L acetonitrile+0.25 mL 99% formic acid; gradient: 0.0 min 90% A.fwdarw.0.3 min 90% A.fwdarw.1.7 min 5% A.fwdarw.3.0 min 5% A; oven: 50° C.; flow rate: 1.20 mL/min; UV detection: 205-305 nm.

(9) Method 4 (Preparative HPLC):

(10) Column: Chromatorex C18 10 μm, 125 mm×30 mm; eluent A: water+0.05% TFA, eluent B: acetonitrile+0.05% TFA; gradient: 20% B.fwdarw.45% B, 45% B isocratic, 45% B.fwdarw.80% B; column temperature: room temperature; flow rate: 50 mL/min; UV detection: 210 nm.

(11) Starting Materials and Intermediates:

Example 1A

Methyl{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}acetate

(12) ##STR00015##

(13) Under argon, potassium tert-butoxide (9.118 g, 81.26 mmol) was added portionwise at room temperature to a solution of 5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (Example 5A in WO 2011/104322-A1; 20 g, 65.01 mmol) in THF (40 ml). To this solution was added methyl bromoacetate (10.939 g, 71.51 mmol), and the mixture was stirred at room temperature overnight. The reaction mixture was then diluted with water and extracted with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. 16.4 g (30.23 mmol) of the desired compound were obtained (46.5% yield, 70% purity).

(14) LC/MS [method 1]: R.sub.t=0.90 min; MS [ESIpos]: m/z=380 (M+H).sup.+

(15) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 3.70 (s, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.19-4.33 (m, 1H), 4.72 (s, 2H), 6.92 (d, 1H), 7.60-7.69 (m, 2H), 7.73-7.81 (m, 2H).

(16) The title compound can also be synthesized via the procedure described in WO 2011/104322-A1 (Example 7A).

Example 2A

2-{3-(4-Chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}acetohydrazide

(17) ##STR00016##

(18) 7.2 g (18.96 mmol) of methyl{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}acetate were dissolved in 60 ml of absolute ethanol. To this solution were added 2.088 g (41.71 mmol) of hydrazine hydrate, and the mixture was stirred under reflux for 5 h and then at room temperature overnight. The resulting mixture was partially concentrated in vacuo and then diluted with water and extracted with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was dissolved in dichloromethane, and after crystallization the white solid was filtered off and dried under high vacuum. 7.02 g (18.49 mmol) of the desired compound were obtained (97.5% yield).

(19) LC/MS [method 1]: R.sub.t=0.73 min; MS [ESIpos]: m/z=380 (M+H).sup.+

(20) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 3.82 (dd, 1H), 3.96 (dd, 1H), 4.24-4.34 (m, 3H), 4.38 (d, 2H), 6.90 (d, 1H), 7.61-7.66 (m, 2H), 7.73-7.78 (m, 2H), 9.23 (t, 1H).

Example 3A

5-(4-Chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(21) ##STR00017##

(22) Under argon, sodium ethoxide (2.987 g, 42.14 mmol, 96% purity) was added portionwise at room temperature to a solution of 2-{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}acetohydrazide (8.0 g, 21.07 mmol) and 2-hydroxyacetamidine hydrochloride (2.329 g, 21.07 mmol) in DMF (200 ml). The reaction mixture was stirred at 100° C. overnight. After cooling, the reaction mixture was partially concentrated in vacuo and then diluted with ethyl acetate. The resulting mixture was washed with water, and after phase separation, the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The resulting solid was dried under high vacuum to give 8.69 g (89% purity, 18.47 mmol) of the desired compound which was used without further purification (−88% yield).

(23) LC/MS [method 1]: R.sub.t=0.74 min; MS [ESIpos]: m/z=419 (M+H).sup.+

(24) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 3.83 (dd, 1H), 3.98 (dd, 1H), 4.24-4.36 (m, 1H), 4.53 (br. s, 2H), 4.96 (br. s, 2H), 5.64 (br. s, 1H), 6.91 (d, 1H), 7.58-7.67 (m, 2H), 7.72-7.78 (m, 2H), 13.75 (br. s, 1H).

Example 4A

5-(4-Chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(25) ##STR00018##

(26) Under argon, sodium ethoxide (1.531 g, 21.59 mmol, 96% purity) was added portionwise at room temperature to a solution of 2-{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}acetohydrazide (4.1 g, 10.80 mmol) and 2-hydroxypropanimidamide hydrochloride (1.480 g, 11.88 mmol) in DMF (110 ml). The reaction mixture was stirred at 120° C. for 4.5 h. After cooling, the reaction mixture was partially concentrated in vacuo and then diluted with ethyl acetate. The resulting mixture was washed with water, and after phase separation, the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The resulting solid was dried under high vacuum to give 4.90 g (92% purity, 10.42 mmol) of the desired compound as a mixture of diastereomers which was used without further purification.

(27) LC/MS [method 1]: R.sub.t=0.82 min; MS [ESIpos]: m/z=433 (M+H).sup.+

(28) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.39 (d, 3H), 3.79-3.88 (m, 1H), 3.93-4.02 (m, 1H), 4.24-4.36 (m, 1H), 4.80 (quin, 1H), 4.89-5.00 (m, 2H), 5.73 (d, 1H), 6.93 (d, 1H), 7.58-7.65 (m, 2H), 7.70-7.77 (d, 2H), 13.68 (s, 1H).

Example 5A

5-(4-Chlorophenyl)-2-({5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(29) ##STR00019##

(30) The title compound was synthesized analogously to Example 4A starting from (2S)-2-hydroxypropanimidamide hydrochloride (1.1 eq.), using only 1.1 eq. of sodium ethoxide as base (reaction temperature: 100° C.).

(31) LC/MS [method 1]: R.sub.t=0.81 min; MS [ESIpos]: m/z=433 (M+H).sup.+.

(32) (2S)-2-hydroxypropanimidamide hydrochloride, which is commercially available, was also synthesized from (2S)-2-hydroxypropanamide [L-(−)-lactamide] via the procedure described in WO 00/59510-A1 (Preparation 11, Step A) and WO 2013/138860-A1 (Intermediate to Precursor 82, p. 101-102).

Example 6A

5-(4-Chlorophenyl)-2-({5-[(1R)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(33) ##STR00020##

(34) The title compound was synthesized analogously to Example 4A starting from (2R)-2-hydroxypropanimidamide hydrochloride (1.1 eq.), using only 1.1 eq. of sodium ethoxide as base (reaction temperature: 100° C.).

(35) LC/MS [method 3]: R.sub.t=1.00 min; MS [ESIpos]: m/z=433 (M+H).sup.+.

(36) (2R)-2-hydroxypropanimidamide hydrochloride, which is commercially available, was also synthesized from (2R)-2-hydroxypropanamide [R-(+)-lactamide] via the procedure described in WO 00/59510-A1 (Preparation 11, Step A) and WO 2013/138860-A1 (Intermediate to Precursor 82, p. 101-102).

Preparation Examples

(37) Example compounds bearing a 1-hydroxyethyl substituent [R.sup.1 in formula (I)=CH.sub.3] which in the following are termed “diastereomer 1” and “diastereomer 2”, respectively, represent pairs of separated diastereomers whose absolute configuration with regard to the 1-hydroxyethyl moiety (1R or 1S) had not been determined.

(38) Diastereomeric excess (d.e.) values were determined in the usual way by analysis of HPLC peak areas according to the following formula:

(39) $d.e.=.Math.\frac{\mathrm{diastereomer}1\left(\%\mathrm{peak}\mathrm{area}\right)-\mathrm{diastereomer}2\left(\%\mathrm{peak}\mathrm{area}\right)}{\mathrm{diastereomer}1\left(\%\mathrm{peak}\mathrm{area}\right)+\mathrm{diastereomer}2\left(\%\mathrm{peak}\mathrm{area}\right)}.Math.\times 100\%.$

Example 1

5-(4-Chlorophenyl)-2-{[1-(3-chlorophenyl)-5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(40) ##STR00021##

(41) To a solution of 5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (400 mg, 0.96 mmol) in pyridine (12 ml) were added (3-chlorophenyl)boronic acid (298.74 mg, 1.91 mmol) and copper(II) acetate (347 mg, 1.91 mmol). The reaction mixture was stirred at room temperature for 5 days, after which extra boronic acid (74.7 mg, 0.48 mmol, 0.5 eq.) was added due to incomplete conversion. After stirring for two additional days, the reaction mixture was diluted with MTBE and then quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (113 mg, 0.21 mmol) was obtained (yield 22.4%).

(42) LC/MS [method 2]: R.sub.t=3.07 min; MS [ESIpos]: m/z=529 (M+H).sup.+

(43) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.58 (s, 2H), 5.08 (s, 2H), 6.90 (br. d, 1H), 7.55-7.64 (m, 4H), 7.65-7.69 (m, 1H), 7.73-7.78 (m, 2H), 7.78-7.81 (m, 1H).

Example 2

5-(4-Chlorophenyl)-2-{[1-(3-fluorophenyl)-5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(44) ##STR00022##

(45) To a solution of 5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (100 mg, 0.24 mmol) in pyridine (3 ml) were added (3-fluorophenyl)boronic acid (66.83 mg, 0.48 mmol) and copper(II) acetate (86.75 mg, 0.48 mmol). The reaction mixture was stirred at room temperature for 5 days, after which extra boronic acid (16.72 mg, 0.12 mmol, 0.5 eq.) was added due to incomplete conversion. After stirring for two additional days, the reaction mixture was diluted with MTBE and then quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (25 mg, 0.05 mmol) was obtained (yield 20.2%).

(46) LC/MS [method 2]: R.sub.t=2.87 min; MS [ESIpos]: m/z=513 (M+H).sup.+

(47) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.59 (s, 2H), 5.08 (s, 2H), 6.90 (br. d, 1H), 7.37 (td, 1H), 7.51-7.66 (m, 5H), 7.72-7.79 (m, 2H).

Example 3

5-(4-Chlorophenyl)-2-{[5-(hydroxymethyl)-1-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(48) ##STR00023##

(49) To a solution of 5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (300 mg, 0.72 mmol) in pyridine (9 ml) were added (2-methoxyphenyl)boronic acid (217.72 mg, 1.43 mmol) and copper(II) acetate (260.25 mg, 1.43 mmol). The reaction mixture was stirred at room temperature for 5 days, after which extra boronic acid (54.4 mg, 0.36 mmol, 0.5 eq.) was added due to incomplete conversion. After stirring for two additional days, the reaction mixture was diluted with MTBE and then quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (73 mg, 0.14 mmol) was obtained (yield 22.4%, 98% purity).

(50) LC/MS [method 3]: R.sub.t=1.18 min; MS [ESIpos]: m/z=525 (M+H).sup.+

(51) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 3.77 (s, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.31 (br. s, 1H), 4.35 (s, 2H), 5.04 (s, 2H), 6.91 (br. s, 1H), 7.09 (t, 1H), 7.25 (d, 1H), 7.37 (dd, 1H), 7.52 (td, 1H), 7.63 (d, 2H), 7.76 (d, 2H).

Example 4

5-(4-Chlorophenyl)-2-{[1-(2-chlorophenyl)-5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(52) ##STR00024##

(53) To a solution of 5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (3 g, 5.946 mmol, 83% purity) in pyridine (75 ml) were added (2-chlorophenyl)boronic acid (930 mg, 5.946 mmol) and copper(II) acetate (2.16 g, 11.89 mmol). The reaction mixture was stirred at room temperature overnight, after which extra boronic acid (500 mg, 3.20 mmol) was added due to incomplete conversion. The reaction mixture was further stirred at room temperature for 9 days. Over this time, three additional portions of boronic acid (1.5 g in total, 9.6 mmol) were added. After this, the reaction mixture was diluted with MTBE and then quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (1.44 g, 2.72 mmol) was obtained (yield 45.7%).

(54) LC/MS [method 2]: R.sub.t=2.79 min; MS [ESIpos]: m/z=529 (M+H).sup.+

(55) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 3.85 (dd, 1H), 4.01 (dd, 1H), 4.31 (br. s, 1H), 4.41 (s, 2H), 5.07 (s, 2H), 6.91 (d, 1H), 7.49-7.67 (m, 5H), 7.68-7.80 (m, 3H).

Example 5

5-(4-Chlorophenyl)-2-{[5-(hydroxymethyl)-1-(2-methylphenyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(56) ##STR00025##

(57) To a solution of 5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (400 mg, 0.96 mmol) in pyridine (12 ml) were added (2-methylphenyl)boronic acid (259.7 mg, 1.91 mmol) and copper(II) acetate (347 mg, 1.91 mmol). The reaction mixture was stirred at room temperature for 5 days, after which extra boronic acid (64.9 mg, 0.48 mmol, 0.5 eq.) was added due to incomplete conversion. After stirring for two additional days, the reaction mixture was diluted with MTBE and then quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (58 mg, 0.11 mmol) was obtained (yield 11.9%).

(58) LC/MS [method 2]: R.sub.t=2.85 min; MS [ESIpos]: m/z=509 (M+H).sup.+

(59) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 2.01 (s, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.34 (s, 2H), 5.07 (s, 2H), 6.90 (br. s, 1H), 7.32-7.50 (m, 4H), 7.62 (br. d, 2H), 7.75 (br. d, 2H).

Example 6

2-{[1-(3-Chloro-5-fluorophenyl)-5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(60) ##STR00026##

(61) To a solution of 5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (400 mg, 0.96 mmol) in pyridine (12 ml) were added (3-chloro-5-fluorophenyl)boronic acid (333.1 mg, 1.91 mmol) and copper(II) acetate (347 mg, 1.91 mmol). The reaction mixture was stirred at room temperature for 5 days, after which extra boronic acid (83.2 mg, 0.48 mmol, 0.5 eq.) was added due to incomplete conversion. After stirring for two additional days, the reaction mixture was diluted with MTBE and then quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (91 mg, 0.17 mmol) was obtained (yield 17.4%).

(62) LC/MS [method 3]: R.sub.t=1.31 min; MS [ESIpos]: m/z=547 (M+H).sup.+

(63) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.63 (s, 2H), 5.08 (s, 2H), 6.90 (br. s, 1H), 7.59-7.66 (m, 4H), 7.67-7.71 (m, 1H), 7.72-7.78 (m, 2H).

Example 7

5-(4-Chlorophenyl)-2-{[1-(3,5-difluorophenyl)-5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(64) ##STR00027##

(65) The title compound was prepared analogously to Example 1 starting from 360 mg (0.86 mmol) of 5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one. 61 mg (0.11 mmol) of the desired compound were obtained (13.4% yield).

(66) LC/MS [method 3]: R.sub.t=1.25 min; MS [ESIpos]: m/z=531 (M+H).sup.+

(67) .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ [ppm] 3.86 (dd, 1H), 4.01 (dd, 1H), 4.25-4.38 (m, 1H), 4.64 (d, 2H), 5.08 (s, 2H), 5.94 (t, 1H), 6.92 (d, 1H), 7.46 (tt, 1H), 7.49-7.55 (m, 2H), 7.62 (br. d, 2H), 7.76 (br. d, 2H).

Example 8

5-(4-Chlorophenyl)-2-({5-(hydroxymethyl)-1-[2-(trifluoromethyl)phenyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(68) ##STR00028##

(69) The title compound was prepared analogously to Example 1 starting from 500 mg (1.19 mmol) of 5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one. 104 mg (0.18 mmol) of the desired compound were obtained (15.5% yield).

(70) LC/MS [method 3]: R.sub.t=1.24 min; MS [ESIpos]: m/z=563 (M+H).sup.+

(71) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.39 (s, 2H), 5.01-5.12 (m, 2H), 5.53 (br. s, 1H), 6.92 (d, 1H), 7.63 (d, 2H), 7.70 (d, 1H), 7.75 (d, 2H), 7.78-7.91 (m, 2H), 7.97 (d, 1H).

Example 9

2-{[1-(2-Chloro-5-fluorophenyl)-5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(72) ##STR00029##

(73) The title compound was prepared analogously to Example 1 starting from 500 mg (1.19 mmol) of 5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one. 8.7 mg (0.02 mmol) of the desired compound were obtained (1.3% yield, 95% purity).

(74) LC/MS [method 3]: R.sub.t=1.23 min; MS [ESIpos]: m/z=547 (M+H).sup.+

(75) .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ [ppm] 3.86 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.46 (s, 2H), 5.03-5.11 (m, 2H), 6.92 (br. d, 1H), 7.53 (td, 1H), 7.61-7.65 (m, 2H), 7.68 (dd, 1H), 7.73-7.80 (m, 3H).

Example 10

5-(4-Chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(76) ##STR00030##

(77) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (500 mg, 1.16 mmol) in pyridine (15 ml) were added (2-methoxyphenyl)boronic acid (351.1 mg, 2.31 mmol) and copper(II) acetate (419.7 mg, 2.31 mmol). The reaction mixture was stirred at room temperature for 5 days, after which extra boronic acid (87 mg, 0.58 mmol, 0.5 eq.) was added due to incomplete conversion. After stirring for two additional days, the reaction mixture was concentrated in vacuo, then diluted with MTBE and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (132 mg, 0.22 mmol) was obtained as a mixture of diastereomers (yield 19.1%, 90% purity).

(78) LC/MS [method 2]: R.sub.t=2.87 min; MS [ESIpos]: m/z=539 (M+H).sup.+

(79) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.36 (d, 3H), 3.76 (s, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.47-4.61 (m, 1H), 4.98-5.10 (m, 2H), 6.90 (br. s, 1H), 7.09 (td, 1H), 7.24 (dd, 1H), 7.35 (dd, 1H), 7.49-7.55 (m, 1H), 7.60-7.65 (m, 2H), 7.73-7.79 (m, 2H).

(80) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 128 mg dissolved in 10 ml methanol; injection volume: 0.3 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/methanol 70:30; flow rate: 80 ml/min; temperature: 40° C.; UV detection: 210 nm]. After separation, 43.6 mg of diastereomer 1 (Example 11), which eluted first, and 45.1 mg of diastereomer 2 (Example 12), which eluted later, were isolated.

Example 11

5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(81) Analytical chiral HPLC (SFC): R.sub.t=3.08 min, d.e.=100% [column: Daicel Chiralcel OX-3 250×4 mm; eluent: carbon dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV detection: 220 nm].

(82) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.36 (d, 3H), 3.76 (s, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.52 (quin, 1H), 4.96-5.11 (m, 2H), 5.37 (d, 1H), 6.90 (d, 1H), 7.09 (td, 1H), 7.24 (d, 1H), 7.35 (dd, 1H), 7.52 (td, 1H), 7.60-7.65 (m, 2H), 7.73-7.79 (m, 2H).

Example 12

5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(83) Analytical chiral HPLC (SFC): R.sub.t=3.38 min, d.e.=91.1% [column: Daicel Chiralcel® OX-3 250×4 mm; eluent: carbon dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV detection: 220 nm].

(84) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.36 (d, 3H), 3.76 (s, 3H), 3.84 (dd, 1H), 4.00 (dd, 1H), 4.31 (br. s, 1H), 4.52 (quin, 1H), 4.99-5.09 (m, 2H), 5.37 (d, 1H), 6.91 (d, 1H), 7.09 (td, 1H), 7.24 (d, 1H), 7.35 (dd, 1H), 7.52 (td, 1H), 7.60-7.65 (m, 2H), 7.74-7.79 (m, 2H).

Example 13

2-({1-(3-Chloro-5-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(85) ##STR00031##

(86) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (430 mg, 0.99 mmol) in pyridine (12.5 ml) were added (3-chloro-5-fluorophenyl)boronic acid (346.49 mg, 1.99 mmol) and copper(II) acetate (360.9 mg, 1.99 mmol). The reaction mixture was stirred at room temperature for 5 days, after which extra boronic acid (86.7 mg, 0.497 mmol, 0.5 eq.) was added due to incomplete conversion. After stirring for two additional days, the reaction mixture was concentrated in vacuo, then diluted with MTBE and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (148 mg, 0.26 mmol) was obtained as a mixture of diastereomers (yield 26.5%).

(87) LC/MS [method 2]: R.sub.t=3.28 min; MS [ESIpos]: m/z=561 (M+H).sup.+

(88) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.48 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.29 (br. s, 1H), 4.83-4.91 (m, 1H), 5.01-5.13 (m, 2H), 6.89 (br. s, 1H), 7.55-7.70 (m, 5H), 7.72-7.78 (m, 2H).

(89) The two diastereomers were separated by preparative chiral HPLC (SFC) [sample preparation: 143 mg dissolved in 15 ml methanol; injection volume: 0.5 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: carbon dioxide/methanol 80:20; flow rate: 80 ml/min; temperature: 40° C.; UV detection: 210 nm]. After separation, 70 mg of diastereomer 1 (Example 14), which eluted first, and 60 mg of diastereomer 2 (Example 15), which eluted later, were isolated.

Example 14

2-{[1-(3-Chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(90) Analytical chiral HPLC (SFC): R.sub.t=4.45 min, d.e.=100% [column: Daicel Chiralcel OX-3 250×4 mm; eluent: carbon dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV detection: 220 nm].

(91) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.48 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.36 (m, 1H), 4.87 (quin, 1H), 5.07 (s, 2H), 5.83 (d, 1H), 6.89 (d, 1H), 7.58-7.69 (m, 5H), 7.71-7.79 (m, 2H).

Example 15

2-{[1-(3-Chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(92) Analytical chiral HPLC (SFC): R.sub.t=4.80 min, d.e.=100% [column: Daicel Chiralcel OX-3 250×4 mm; eluent: carbon dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV detection: 220 nm].

(93) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.48 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.23-4.36 (m, 1H), 4.87 (quin, 1H), 5.02-5.12 (m, 2H), 5.83 (d, 1H), 6.89 (d, 1H), 7.57-7.70 (m, 5H), 7.71-7.79 (m, 2H).

Example 16

5-(4-Chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1-phenyl-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(94) ##STR00032##

(95) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (250 mg, 0.462 mmol, 80% purity) in pyridine (6 ml) were added phenylboronic acid (112.69 mg, 0.92 mmol) and copper-(II) acetate (167.9 mg, 0.92 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for 4 days, after which extra boronic acid (28.2 mg, 0.23 mmol, 0.5 eq.)

(96) was added due to incomplete conversion. After further 4 h at 60° C. followed by stirring at room temperature for two additional days, the reaction mixture was concentrated in vacuo, then diluted with ethyl acetate and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (42.5 mg, 0.08 mmol) was obtained as a mixture of diastereomers (yield 18.1%).

(97) LC/MS [method 3]: R.sub.t=1.21 min; MS [ESIpos]: m/z=509 (M+H).sup.+

(98) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.45 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.77 (q, 1H), 4.99-5.13 (m, 2H), 6.90 (br. s, 1H), 7.47-7.66 (m, 7H), 7.72-7.79 (m, 2H).

(99) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 38.9 mg dissolved in 1 ml ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 75:25; flow rate: 15 ml/min; temperature: 30° C.; UV detection: 220 nm]. After separation, 13 mg of diastereomer 1 (Example 17), which eluted first, and 14 mg of diastereomer 2 (Example 18), which eluted later, were isolated.

Example 17

5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-phenyl-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(100) Analytical chiral HPLC: R.sub.t=8.18 min, d.e.=100% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(101) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.45 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.77 (quin, 1H), 4.98-5.14 (m, 2H), 5.68 (d, 1H), 6.89 (d, 1H), 7.48-7.65 (m, 7H), 7.72-7.80 (m, 2H).

Example 18

5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-phenyl-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(102) Analytical chiral HPLC: R.sub.t=11.40 min, d.e.=100% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(103) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.45 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.77 (quin, 1H), 5.07 (s, 2H), 5.68 (d, 1H), 6.90 (d, 1H), 7.47-7.65 (m, 7H), 7.72-7.79 (m, 2H).

Example 19

5-(4-Chlorophenyl)-2-({1-[3-(difluoromethyl)phenyl]-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(104) ##STR00033##

(105) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (400 mg, 0.74 mmol, 80% purity) in pyridine (9.6 ml) were added [3-(difluoromethyl)phenyl]boronic acid (254.26 mg, 1.48 mmol) and copper(II) acetate (268.6 mg, 1.48 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for 5 days. The resulting reaction mixture was concentrated in vacuo, then diluted with ethyl acetate and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (47 mg, 0.08 mmol) was obtained as a mixture of diastereomers (yield 11.3%).

(106) LC/MS [method 1]: R.sub.t=1.04 min; MS [ESIpos]: m/z=559 (M+H).sup.+

(107) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.47 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.36 (m, 1H), 4.81 (q, 1H), 5.02-5.13 (m, 2H), 5.74 (br. s, 1H), 6.89 (br. s, 1H), 7.14 (t, 1H), 7.59-7.65 (m, 2H), 7.69-7.78 (m, 4H), 7.81-7.87 (m, 2H).

(108) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 45 mg dissolved in 1 ml ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 75:25; flow rate: 15 ml/min; temperature: 30° C.; UV detection: 220 nm]. After separation, 20 mg of diastereomer 1 (Example 20), which eluted first, and 20 mg of diastereomer 2 (Example 21), which eluted later, were isolated.

Example 20

5-(4-Chlorophenyl)-2-({1-[3-(difluoromethyl)phenyl]-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(109) Analytical chiral HPLC: R.sub.t=6.72 min, d.e.=99% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(110) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.47 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.23-4.36 (m, 1H), 4.81 (quin, 1H), 5.02-5.14 (m, 2H), 5.74 (d, 1H), 6.88 (d, 1H), 7.14 (t, 1H), 7.59-7.64 (m, 2H), 7.69-7.79 (m, 4H), 7.80-7.87 (m, 2H).

Example 21

5-(4-Chlorophenyl)-2-({1-[3-(difluoromethyl)phenyl]-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(111) Analytical chiral HPLC: R.sub.t=9.36 min, d.e.=100% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(112) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.47 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.37 (m, 1H), 4.81 (quin, 1H), 5.08 (s, 2H), 5.74 (d, 1H), 6.89 (d, 1H), 7.13 (t, 1H), 7.58-7.66 (m, 2H), 7.69-7.79 (m, 4H), 7.81-7.87 (m, 2H).

Example 22

5-(4-Chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1-[3-(trifluoromethyl)phenyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(113) ##STR00034##

(114) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (400 mg, 0.74 mmol, 80% purity) in pyridine (9.6 ml) were added [3-(trifluoromethyl)phenyl]boronic acid (280.86 mg, 1.48 mmol) and copper(II) acetate (268.6 mg, 1.48 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for 5 days, after which extra boronic acid (70.2 mg, 0.37 mmol, 0.5 eq.) was added due to incomplete conversion. After further 4 h at 60° C. followed by stirring at room temperature overnight, the reaction mixture was concentrated in vacuo, then diluted with MTBE and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (58.2 mg, 0.10 mmol) was obtained as a mixture of diastereomers (yield 13.6%).

(115) LC/MS [method 1]: R.sub.t=1.06 min; MS [ESIpos]: m/z=577 (M+H).sup.+

(116) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.48 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.83 (q, 1H), 5.02-5.15 (m, 2H), 5.79 (br. s, 1H), 6.85-6.94 (m, 1H), 7.58-7.66 (m, 2H), 7.71-7.85 (m, 3H), 7.86-7.92 (m, 1H), 7.94-8.01 (m, 1H), 8.04 (s, 1H).

(117) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 58 mg dissolved in 2 ml ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 75:25; flow rate: 15 ml/min; temperature: 30° C.; UV detection: 220 nm]. After separation, 19.5 mg of diastereomer 1 (Example 23), which eluted first, and 19.2 mg of diastereomer 2 (Example 24), which eluted later, were isolated.

Example 23

5-(4-Chlorophenyl)-2-({5-(1-hydroxyethyl)-1-[3-(trifluoromethyl)phenyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(118) Analytical chiral HPLC: R.sub.t=4.94 min, d.e.=100% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(119) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.48 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.83 (q, 1H), 5.03-5.13 (m, 2H), 5.79 (br. s, 1H), 6.88 (d, 1H), 7.59-7.65 (m, 2H), 7.72-7.85 (m, 3H), 7.86-7.92 (m, 1H), 7.95-8.01 (m, 1H), 8.04 (br. s, 1H).

Example 24

5-(4-Chlorophenyl)-2-({5-(1-hydroxyethyl)-1-[3-(trifluoromethyl)phenyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(120) Analytical chiral HPLC: R.sub.t=6.13 min, d.e.=98.6% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(121) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.48 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.36 (m, 1H), 4.83 (quin, 1H), 5.09 (s, 2H), 5.78 (d, 1H), 6.89 (d, 1H), 7.62 (d, 2H), 7.71-7.77 (m, 2H), 7.78-7.85 (m, 1H), 7.86-7.92 (m, 1H), 7.94-8.01 (m, 1H), 8.04 (br. s, 1H).

Example 25

5-(4-Chlorophenyl)-2-({1-(3,5-difluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(122) ##STR00035##

(123) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added (3,5-difluorophenyl)boronic acid (291.9 mg, 1.85 mmol) and copper(II) acetate (335.7 mg, 1.85 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for 4 days, after which extra boronic acid (72.98 mg, 0.46 mmol, 0.5 eq.) was added due to incomplete conversion. After further 2 h at 60° C. followed by stirring at room temperature for three additional days, the reaction mixture was concentrated in vacuo, then diluted with ethyl acetate and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (114.3 mg, 0.21 mmol) was obtained as a mixture of diastereomers (yield 22.7%).

(124) LC/MS [method 3]: R.sub.t=1.28 min; MS [ESIpos]: m/z=545 (M+H).sup.+

(125) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.49 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.88 (q, 1H), 5.02-5.13 (m, 2H), 6.89 (br. s, 1H), 7.41-7.54 (m, 3H), 7.62 (d, 2H), 7.75 (d, 2H).

(126) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 110 mg dissolved in 7 ml ethanol/isohexane (1:1); injection volume: 0.6 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 70:30; flow rate: 20 ml/min; temperature: 40° C.; UV detection: 220 nm]. After separation, 42 mg of diastereomer 1 (Example 26), which eluted first, and 44 mg of diastereomer 2 (Example 27), which eluted later, were isolated.

Example 26

5-(4-Chlorophenyl)-2-{[1-(3,5-difluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(127) LC/MS [method 2]: R.sub.t=3.10 min; MS [ESIpos]: m/z=545 (M+H).sup.+

(128) Analytical chiral HPLC: R.sub.t=1.09 min, d.e.=100% [column: Daicel Chiralpack OX-3 3 μm, 50×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; UV detection: 220 nm].

(129) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.49 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.23-4.36 (m, 1H), 4.88 (quin, 1H), 5.02-5.13 (m, 2H), 5.84 (d, 1H), 6.89 (d, 1H), 7.41-7.54 (m, 3H), 7.59-7.65 (m, 2H), 7.73-7.76 (m, 2H).

Example 27

5-(4-Chlorophenyl)-2-{[1-(3,5-difluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(130) LC/MS [method 2]: R.sub.t=3.09 min; MS [ESIpos]: m/z=545 (M+H).sup.+

(131) Analytical chiral HPLC: R.sub.t=1.28 min, d.e.=99% [column: Daicel Chiralpack OX-3 3 μm, 50×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; UV detection: 220 nm].

(132) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.48 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.36 (m, 1H), 4.88 (quin, 1H), 5.07 (s, 2H), 5.83 (d, 1H), 6.90 (d, 1H), 7.42-7.54 (m, 3H), 7.59-7.65 (m, 2H), 7.72-7.79 (m, 2H).

Example 28

5-(4-Chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1-(3-methylphenyl)-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(133) ##STR00036##

(134) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added (3-methylphenyl)boronic acid (251.32 mg, 1.85 mmol) and copper(II) acetate (335.7 mg, 1.85 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for three days. The resulting reaction mixture was concentrated in vacuo, then diluted with ethyl acetate and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (59.6 mg, 0.11 mmol) was obtained as a mixture of diastereomers (yield 12.3%).

(135) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.44 (d, 3H), 2.38 (s, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.29 (br. s, 1H), 4.76 (q, 1H), 5.01-5.11 (m, 2H), 5.66 (br. s, 1H), 6.90 (t, 1H), 7.29-7.35 (m, 1H), 7.38-7.47 (m, 3H), 7.59-7.65 (m, 2H), 7.72-7.78 (m, 2H).

(136) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 56 mg dissolved in 2 ml ethanol/isohexane (1:1); injection volume: 0.5 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 70:30; flow rate: 15 ml/min; temperature: 25° C.; UV detection: 220 nm]. After separation, 22 mg of diastereomer 1 (Example 29), which eluted first, and 24 mg of diastereomer 2 (Example 30), which eluted later, were isolated.

Example 29

5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(3-methylphenyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(137) Analytical chiral HPLC: R.sub.t=7.97 min, d.e.=100% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(138) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.44 (d, 3H), 2.38 (s, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.36 (m, 1H), 4.76 (quin, 1H), 5.00-5.11 (m, 2H), 5.67 (d, 1H), 6.89 (d, 1H), 7.30-7.35 (m, 1H), 7.38-7.47 (m, 3H), 7.59-7.65 (m, 2H), 7.72-7.78 (m, 2H).

Example 30

5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(3-methylphenyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(139) Analytical chiral HPLC: R.sub.t=11.44 min, d.e.=99.1% [column: LUX Cellulose-4, 5 rim, 250×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(140) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.44 (d, 3H), 2.38 (s, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.36 (m, 1H), 4.77 (quin, 1H), 5.01-5.10 (m, 2H), 5.67 (d, 1H), 6.90 (d, 1H), 7.29-7.35 (m, 1H), 7.38-7.47 (m, 3H), 7.59-7.65 (m, 2H), 7.73-7.78 (m, 2H).

Example 31

5-(4-Chlorophenyl)-2-({1-(2-ethylphenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(141) ##STR00037##

(142) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (600 mg, 1.39 mmol) in pyridine (18 ml) were added (2-ethylphenyl)boronic acid (415.87 mg, 2.77 mmol) and copper(II) acetate (503.6 mg, 2.77 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for three days. The resulting reaction mixture was concentrated in vacuo, then diluted with ethyl acetate and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (69.4 mg, 0.13 mmol) was obtained as a mixture of diastereomers (yield 9.1%).

(143) LC/MS [method 2]: R.sub.t=3.16 min; MS [ESIpos]: m/z=537 (M+H).sup.+

(144) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 0.98 (t, 3H), 1.37 (d, 3H), 2.27 (qd, 2H), 3.84 (dd, 1H), 4.00 (dd, 1H), 4.21-4.37 (m, 1H), 4.52 (q, 1H), 5.00-5.13 (m, 2H), 5.48 (br. s, 1H), 6.90 (dd, 1H), 7.32-7.40 (m, 2H), 7.41-7.54 (m, 2H), 7.58-7.65 (m, 2H), 7.70-7.77 (m, 2H).

(145) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 65 mg dissolved in 4 ml ethanol/isohexane (1:1); injection volume: 0.5 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 50:50; flow rate: 20 ml/min; temperature: 40° C.; UV detection: 220 nm]. After separation, 25 mg of diastereomer 1 (Example 32), which eluted first, and 25 mg of diastereomer 2 (Example 33), which eluted later, were isolated.

Example 32

5-(4-Chlorophenyl)-2-{[1-(2-ethylphenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(146) LC/MS [method 2]: R.sub.t=3.15 min; MS [ESIpos]: m/z=537 (M+H).sup.+

(147) Analytical chiral HPLC: R.sub.t=0.96 min, d.e.=100% [column: Daicel Chiralpack OX-3 3 μm, 50×4.6 mm; eluent: isohexane/ethanol 50:50; flow rate: 1 ml/min; UV detection: 220 nm].

(148) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 0.98 (t, 3H), 1.37 (d, 3H), 2.27 (qd, 2H), 3.84 (dd, 1H), 4.00 (dd, 1H), 4.23-4.35 (m, 1H), 4.52 (quin, 1H), 5.00-5.12 (m, 2H), 5.48 (d, 1H), 6.89 (d, 1H), 7.32-7.40 (m, 2H), 7.42-7.53 (m, 2H), 7.59-7.66 (m, 2H), 7.71-7.78 (m, 2H).

Example 33

5-(4-Chlorophenyl)-2-{[1-(2-ethylphenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(149) LC/MS [method 2]: R.sub.t=3.15 min; MS [ESIpos]: m/z=537 (M+H).sup.±

(150) Analytical chiral HPLC: R.sub.t=1.09 min, d.e.=100% [column: Daicel Chiralpack OX-3 3 μm, 50×4.6 mm; eluent: isohexane/ethanol 50:50; flow rate: 1 ml/min; UV detection: 220 nm].

(151) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 0.98 (t, 3H), 1.37 (d, 3H), 2.27 (qd, 2H), 3.84 (dd, 1H), 4.00 (dd, 1H), 4.23-4.36 (m, 1H), 4.52 (quin, 1H), 5.07 (s, 2H), 5.48 (d, 1H), 6.90 (d, 1H), 7.32-7.40 (m, 2H), 7.42-7.54 (m, 2H), 7.59-7.66 (m, 2H), 7.71-7.78 (m, 2H).

Example 34

2-({1-(2-Chloro-4-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(152) ##STR00038##

(153) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (600 mg, 1.39 mmol) in pyridine (18 ml) were added (2-chloro-4-fluorophenyl)boronic acid (483 mg, 2.77 mmol) and copper(II) acetate (503.6 mg, 2.77 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for 5 days, after which extra boronic acid (242 mg, 1.39 mmol) was added due to incomplete conversion. The reaction mixture was further stirred at room temperature for 4 days. Over this time, two additional portions of boronic acid (483 mg in total, 2.77 mmol) were added. After this, the reaction mixture was concentrated in vacuo, then diluted with MTBE and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (107 mg, 0.19 mmol) was obtained as a mixture of diastereomers (yield 13.7%).

(154) LC/MS [method 2]: R.sub.t=3.02 min; MS [ESIpos]: m/z=561 (M+H).sup.+

(155) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.38 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.36 (m, 1H), 4.57-4.67 (m, 1H), 5.00-5.12 (m, 2H), 6.90 (br. s, 1H), 7.42 (td, 1H), 7.57-7.79 (m, 6H).

(156) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 104 mg dissolved in 5 ml ethanol/isohexane (1:1); injection volume: 0.5 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 70:30; flow rate: 20 ml/min; temperature: 40° C.; UV detection: 220 nm]. After separation, 56 mg of diastereomer 1 (Example 35), which eluted first, and 29 mg of diastereomer 2 (Example 36), which eluted later, were isolated.

Example 35

2-{[1-(2-Chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(157) Analytical chiral HPLC: R.sub.t=1.36 min, d.e.=100% [column: Daicel Chiralpack OX-3 3 μm, 50×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; UV detection: 220 nm].

(158) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.38 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.36 (m, 1H), 4.61 (quin, 1H), 5.01-5.11 (m, 2H), 5.51 (d, 1H), 6.89 (d, 1H), 7.42 (td, 1H), 7.60-7.65 (m, 2H), 7.66-7.72 (m, 1H), 7.72-7.78 (m, 3H).

Example 36

2-{[1-(2-Chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(159) Analytical chiral HPLC: R.sub.t=1.72 min, d.e.=100% [column: Daicel Chiralpack OX-3 3 μm, 50×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; UV detection: 220 nm].

(160) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.38 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.37 (m, 1H), 4.61 (quin, 1H), 5.06 (s, 2H), 5.51 (d, 1H), 6.90 (d, 1H), 7.42 (td, 1H), 7.60-7.65 (m, 2H), 7.66-7.72 (m, 1H), 7.72-7.78 (m, 3H).

Example 37

5-(4-Chlorophenyl)-2-({1-(5-fluoro-2-methoxyphenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(161) ##STR00039##

(162) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (600 mg, 1.39 mmol) in pyridine (18 ml) were added (5-fluoro-2-methoxyphenyl)boronic acid (471.22 mg, 2.77 mmol) and copper(II) acetate (503.6 mg, 2.77 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for three days. The resulting reaction mixture was concentrated in vacuo, then diluted with ethyl acetate and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (62.2 mg, 0.11 mmol) was obtained as a mixture of diastereomers (yield 8.1%).

(163) LC/MS [method 2]: R.sub.t=2.93 min; MS [ESIpos]: m/z=557 (M+H).sup.+

(164) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.37 (d, 3H), 3.75 (s, 3H), 3.80-3.89 (m, 1H), 4.00 (dd, 1H), 4.24-4.36 (m, 1H), 4.52-4.62 (m, 1H), 4.99-5.04 (m, 2H), 6.90 (t, 1H), 7.26 (dd, 1H), 7.33 (dd, 1H), 7.37-7.44 (m, 1H), 7.60-7.65 (m, 2H), 7.73-7.78 (m, 2H).

(165) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 55.4 mg dissolved in 6 ml ethanol/isohexane (1:1); injection volume: 2 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 80:20; flow rate: 20 ml/min; temperature: 40° C.; UV detection: 220 nm]. After separation, 23 mg of diastereomer 1 (Example 38), which eluted first, and 21 mg of diastereomer 2 (Example 39), which eluted later, were isolated.

Example 38

5-(4-Chlorophenyl)-2-{[1-(5-fluoro-2-methoxyphenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]-methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(166) LC/MS [method 2]: R.sub.t=2.91 min; MS [ESIpos]: m/z=557 (M+H).sup.+

(167) Analytical chiral HPLC: R.sub.t=2.13 min, d.e.=100% [column: Daicel Chiralpack OX-3 3 μm, 50×4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 30° C.; UV detection: 220 nm].

(168) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.37 (d, 3H), 3.75 (s, 3H), 3.81-3.89 (m, 1H), 4.00 (dd, 1H), 4.25-4.35 (m, 1H), 4.53-4.62 (m, 1H), 4.99-5.10 (m, 2H), 5.40 (d, 1H), 6.89 (d, 1H), 7.26 (dd, 1H), 7.33 (dd, 1H), 7.41 (td, 1H), 7.60-7.66 (m, 2H), 7.73-7.79 (m, 2H).

Example 39

5-(4-Chlorophenyl)-2-{[1-(5-fluoro-2-methoxyphenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]-methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(169) LC/MS [method 2]: R.sub.t=2.90 min; MS [ESIpos]: m/z=557 (M+H).sup.+

(170) Analytical chiral HPLC: R.sub.t=2.75 min, d.e.=100% [column: Daicel Chiralpack OX-3 3 μm, 50×4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 30° C.; UV detection: 220 nm].

(171) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.37 (d, 3H), 3.75 (s, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.36 (m, 1H), 4.57 (quin, 1H), 4.99-5.10 (m, 2H), 5.40 (d, 1H), 6.91 (d, 1H), 7.26 (dd, 1H), 7.33 (dd, 1H), 7.41 (td, 1H), 7.60-7.65 (m, 2H), 7.73-7.78 (m, 2H).

Example 40

5-(4-Chlorophenyl)-2-({1-(2-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(172) ##STR00040##

(173) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (600 mg, 1.39 mmol) in pyridine (18 ml) were added (2-fluorophenyl)boronic acid (387.96 mg, 2.77 mmol) and copper(II) acetate (503.6 mg, 2.77 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for 3 days. Over this time, two additional portions of boronic acid (387.96 mg in total, 2.77 mmol) were added. After this, the resulting reaction mixture was concentrated in vacuo, then diluted with ethyl acetate and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (30.1 mg, 0.06 mmol) was obtained as a mixture of diastereomers (yield 4.1%).

(174) LC/MS [method 2]: R.sub.t=2.84 min; MS [ESIpos]: m/z=527 (M+H).sup.+

(175) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.40 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.29 (br. s, 1H), 4.69 (q, 1H), 5.01-5.12 (m, 2H), 6.89 (br. s, 1H), 7.38 (t, 1H), 7.48 (t, 1H), 7.57-7.66 (m, 4H), 7.71-7.80 (m, 2H).

(176) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 26 mg dissolved in 4 ml ethanol/isohexane (1:1); injection volume: 2 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 80:20; flow rate: 25 ml/min; temperature: 40° C.; UV detection: 220 nm]. After separation, 11 mg of diastereomer 1 (Example 41), which eluted first, and 9 mg of diastereomer 2 (Example 42), which eluted later, were isolated.

Example 41

5-(4-Chlorophenyl)-2-{[1-(2-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(177) LC/MS [method 2]: R.sub.t=2.83 min; MS [ESIpos]: m/z=527 (M+H).sup.+

(178) Analytical chiral HPLC: R.sub.t=2.32 min, d.e.=100% [column: Daicel Chiralpack OX-3 3 μm, 50×4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 30° C.; UV detection: 220 nm].

(179) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.40 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.36 (m, 1H), 4.69 (quin, 1H), 5.01-5.12 (m, 2H), 5.53 (d, 1H), 6.89 (d, 1H), 7.38 (t, 1H), 7.48 (t, 1H), 7.57-7.66 (m, 4H), 7.72-7.78 (m, 2H).

Example 42

5-(4-Chlorophenyl)-2-{[1-(2-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(180) LC/MS [method 2]: R.sub.t=2.82 min; MS [ESIpos]: m/z=527 (M+H).sup.+

(181) Analytical chiral HPLC: R.sub.t=3.23 min, d.e.=100% [column: Daicel Chiralpack OX-3 3 μm, 50×4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 30° C.; UV detection: 220 nm].

(182) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.40 (d, 3H), 3.84 (dd, 1H), 4.00 (dd, 1H), 4.24-4.35 (m, 1H), 4.69 (quin, 1H), 5.07 (s, 2H), 5.53 (d, 1H), 6.90 (d, 1H), 7.38 (t, 1H), 7.48 (t, 1H), 7.57-7.67 (m, 4H), 7.72-7.79 (m, 2H).

Example 43

2-({1-(3-Chloro-4-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(183) ##STR00041##

(184) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added (3-chloro-4-fluorophenyl)boronic acid (322.6 mg, 1.85 mmol) and copper(II) acetate (335.7 mg, 1.85 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for 6 days. The resulting reaction mixture was concentrated in vacuo, then diluted with ethyl acetate and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (99.1 mg, 0.18 mmol) was obtained as a mixture of diastereomers (yield 19.1%).

(185) LC/MS [method 3]: R.sub.t=1.31 min; MS [ESIpos]: m/z=561 (M+H).sup.+

(186) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.46 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.80 (q, 1H), 5.01-5.13 (m, 2H), 6.90 (br. s, 1H), 7.59-7.70 (m, 4H), 7.72-7.78 (m, 2H), 7.93 (dd, 1H).

(187) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 97.1 mg dissolved in 3 ml ethanol; injection volume: 0.3 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 25° C.; UV detection: 220 nm]. After separation, 40 mg of diastereomer 1 (Example 44), which eluted first, and 42 mg of diastereomer 2 (Example 45), which eluted later, were isolated.

Example 44

2-{[1-(3-Chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(188) LC/MS [method 2]: R.sub.t=3.22 min; MS [ESIpos]: m/z=561 (M+H).sup.+

(189) Preparative chiral HPLC: R.sub.t=9.97 min, d.e.=100% [column: Daicel Chiralcel OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 25° C.; UV detection: 220 nm].

(190) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.46 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.22-4.36 (m, 1H), 4.80 (quin, 1H), 5.01-5.12 (m, 2H), 5.75 (d, 1H), 6.89 (d, 1H), 7.58-7.70 (m, 4H), 7.71-7.78 (m, 2H), 7.93 (dd, 1H).

Example 45

2-{[1-(3-Chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(191) LC/MS [method 2]: R.sub.t=3.22 min; MS [ESIpos]: m/z=561 (M+H).sup.+

(192) Preparative chiral HPLC: R.sub.t=11.35 min, d.e.=100% [column: Daicel Chiralcel OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 25° C.; UV detection: 220 nm].

(193) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.46 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.25-4.35 (m, 1H), 4.81 (quin, 1H), 5.06 (s, 2H), 5.74 (d, 1H), 6.90 (d, 1H), 7.59-7.70 (m, 4H), 7.73-7.78 (m, 2H), 7.93 (dd, 1H).

Example 46

5-(4-Chlorophenyl)-2-({1-(3,5-dichlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(194) ##STR00042##

(195) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added (3,5-dichlorophenyl)boronic acid (352.73 mg, 1.85 mmol) and copper(II) acetate (335.7 mg, 1.85 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for 6 days. The resulting reaction mixture was concentrated in vacuo, then diluted with ethyl acetate and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (105.5 mg, 0.18 mmol) was obtained as a mixture of diastereomers (yield 19.8%).

(196) LC/MS [method 3]: R.sub.t=1.39 min; MS [ESIpos]: m/z=577 (M+H).sup.+

(197) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.48 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.23-4.36 (m, 1H), 4.82-4.90 (m, 1H), 5.02-5.12 (m, 2H), 6.84-6.94 (m, 1H), 7.59-7.65 (m, 2H), 7.72-7.82 (m, 5H).

(198) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 103.5 mg dissolved in 14 ml ethanol/isohexane (1:1); injection volume: 2 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 80:20; flow rate: 20 ml/min; temperature: 30° C.; UV detection: 220 nm]. After separation, 29.2 mg of diastereomer 1 (Example 47), which eluted first, and 28.9 mg of diastereomer 2 (Example 48), which eluted later, were isolated.

Example 47

5-(4-Chlorophenyl)-2-{[1-(3,5-dichlorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(199) Analytical chiral HPLC: R.sub.t=1.49 min, d.e.=100% [column: Daicel Chiralpack OX-3 3 μm, 50×4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 30° C.; UV detection: 220 nm].

(200) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.48 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.35 (m, 1H), 4.86 (quin, 1H), 5.01-5.12 (m, 2H), 5.82 (d, 1H), 6.88 (d, 1H), 7.58-7.66 (m, 2H), 7.72-7.82 (m, 5H).

Example 48

5-(4-Chlorophenyl)-2-{[1-(3,5-dichlorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(201) Analytical chiral HPLC: R.sub.t=2.02 min, d.e.=99.8% [column: Daicel Chiralpack OX-3 3 μm, 50×4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 30° C.; UV detection: 220 nm].

(202) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.48 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.23-4.36 (m, 1H), 4.86 (quin, 1H), 5.07 (s, 2H), 5.82 (d, 1H), 6.89 (d, 1H), 7.59-7.65 (m, 2H), 7.72-7.81 (m, 5H).

Example 49

5-(4-Chlorophenyl)-2-({1-(2,5-dichlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(203) ##STR00043##

(204) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added (2,5-dichlorophenyl)boronic acid (352.73 mg, 1.85 mmol) and copper(II) acetate (335.75 mg, 1.85 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for three days, after which extra boronic acid (100 mg, 0.52 mmol) was added due to incomplete conversion. The reaction mixture was further stirred at room temperature for 6 days. Over this time, another portion of boronic acid (100 mg, 0.52 mmol) was added. After this, the resulting reaction mixture was concentrated in vacuo, then diluted with ethyl acetate and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (52.8 mg, 0.09 mmol, 97% purity) was obtained as a mixture of diastereomers (yield 9.6%).

(205) LC/MS [method 3]: R.sub.t=1.33 min; MS [ESIpos]: m/z=577 (M+H).sup.+

(206) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.40 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.35 (m, 1H), 4.63-4.72 (m, 1H), 5.01-5.12 (m, 2H), 6.89 (br. s, 1H), 7.60-7.65 (m, 2H), 7.67-7.78 (m, 4H), 7.81 (br. d, 1H).

(207) The two diastereomers were separated by preparative chiral HPLC (SFC) [sample preparation: 50 mg dissolved in 10 ml methanol; injection volume: 0.5 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: carbon dioxide/methanol 82:18; flow rate: 80 ml/min; temperature: 40° C.; UV detection: 210 nm]. After separation, 20.3 mg of diastereomer 1 (Example 50), which eluted first, and 24.1 mg of diastereomer 2 (Example 51), which eluted later, were isolated.

Example 50

5-(4-Chlorophenyl)-2-{[1-(2,5-dichlorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(208) LC/MS [method 3]: R.sub.t=1.30 min; MS [ESIpos]: m/z=577 (M+H).sup.+

(209) Analytical chiral HPLC (SFC): R.sub.t=2.87 min, d.e.=100% [column: Daicel Chiralcel OX-3, 250×4 mm; eluent: carbon dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV detection: 220 nm].

(210) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.40 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.36 (m, 1H), 4.67 (quin, 1H), 5.01-5.12 (m, 2H), 5.53 (d, 1H), 6.89 (d, 1H), 7.60-7.65 (m, 2H), 7.67-7.78 (m, 4H), 7.81 (br. d, 1H).

Example 51

5-(4-Chlorophenyl)-2-{[1-(2,5-dichlorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(211) LC/MS [method 3]: R.sub.t=1.30 min; MS [ESIpos]: m/z=577 (M+H).sup.+

(212) Analytical chiral HPLC (SFC): R.sub.t=3.11 min, d.e.=100% [column: Daicel Chiralcel OX-3, 250×4 mm; eluent: carbon dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV detection: 220 nm].

(213) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.40 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.35 (m, 1H), 4.67 (quin, 1H), 5.01-5.12 (m, 2H), 5.53 (d, 1H), 6.89 (d, 1H), 7.59-7.65 (m, 2H), 7.66-7.78 (m, 4H), 7.81 (br. d, 1H).

Example 52

2-({1-(3-Chloro-2-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(214) ##STR00044##

(215) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added (3-chloro-2-fluorophenyl)boronic acid (322.31 mg, 1.85 mmol) and copper(II) acetate (335.75 mg, 1.85 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for 12 days. Over this time, extra boronic acid (322.31 mg in total, 1.85 mmol) was added portionwise in a daily fashion. After this, the resulting reaction mixture was concentrated in vacuo, then diluted with ethyl acetate and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (15.9 mg, 0.03 mmol, 97% purity) was obtained as a mixture of diastereomers (yield 3%).

(216) LC/MS [method 2]: R.sub.t=3.12 min; MS [ESIpos]: m/z=561 (M+H).sup.+

(217) .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ [ppm] 1.42 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.35 (m, 1H), 4.75 (q, 1H), 5.02-5.12 (m, 2H), 5.57 (br. s, 1H), 6.89 (br. d, 1H), 7.40 (td, 1H), 7.60-7.65 (m, 3H), 7.72-7.77 (m, 2H), 7.81 (ddd, 1H).

(218) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 14 mg dissolved in 1 ml ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 25° C.; UV detection: 220 nm]. After separation, 6 mg of diastereomer 1 (Example 53), which eluted first, and 6 mg of diastereomer 2 (Example 54), which eluted later, were isolated.

Example 53

2-{[1-(3-Chloro-2-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(219) LC/MS [method 2]: R.sub.t=3.14 min; MS [ESIpos]: m/z=561 (M+H).sup.+

(220) Analytical chiral HPLC: R.sub.t=5.49 min, d.e.=100% [column: Daicel Chiralcel OX-H 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(221) .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ [ppm] 1.42 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.34 (m, 1H), 4.75 (quin, 1H), 5.03-5.11 (m, 2H), 5.57 (d, 1H), 6.89 (d, 1H), 7.40 (td, 1H), 7.60-7.65 (m, 3H), 7.73-7.77 (m, 2H), 7.81 (ddd, 1H).

Example 54

2-{[1-(3-Chloro-2-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(222) LC/MS [method 2]: R.sub.t=3.13 min; MS [ESIpos]: m/z=561 (M+H).sup.+

(223) Analytical chiral HPLC: R.sub.t=6.16 min, d.e.=100% [column: Daicel Chiralcel OX-H 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(224) .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ [ppm] 1.42 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.25-4.34 (m, 1H), 4.75 (quin, 1H), 5.07 (s, 2H), 5.57 (d, 1H), 6.89 (d, 1H), 7.40 (td, 1H), 7.60-7.65 (m, 3H), 7.73-7.77 (m, 2H), 7.81 (ddd, 1H).

Example 55

5-(4-Chlorophenyl)-2-({1-[3-(difluoromethoxy)phenyl]-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(225) ##STR00045##

(226) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added [3-(difluoromethoxy)phenyl]boronic acid (347.40 mg, 1.85 mmol) and copper(II) acetate (335.75 mg, 1.85 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for 6 days, after which extra boronic acid (100 mg, 0.53 mmol) was added due to incomplete conversion. The reaction mixture was stirred at room temperature for two additional days. The resulting reaction mixture was concentrated in vacuo, then diluted with ethyl acetate and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (60.3 mg, 0.10 mmol) was obtained as a mixture of diastereomers (yield 11.4%).

(227) LC/MS [method 3]: R.sub.t=1.28 min; MS [ESIpos]: m/z=575 (M+H).sup.+

(228) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.47 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.25-4.35 (m, 1H), 4.78-4.85 (m, 1H), 5.03-5.12 (m, 2H), 6.89 (br. s, 1H), 7.33 (t, 1H), 7.31-7.35 (m, 1H), 7.48-7.56 (m, 2H), 7.59-7.65 (m, 3H), 7.72-7.78 (m, 2H).

(229) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 58 mg dissolved in 2 ml ethanol; injection volume: 0.7 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 35° C.; UV detection: 220 nm]. After separation, 20.7 mg of diastereomer 1 (Example 56), which eluted first, and 17.7 mg of diastereomer 2 (Example 57), which eluted later, were isolated.

Example 56

5-(4-Chlorophenyl)-2-({1-[3-(difluoromethoxy)phenyl]-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(230) Analytical chiral HPLC: R.sub.t=5.57 min, d.e.=98.7% [column: Daicel Chiralcel OX-H 5, 250×4.6 mm; eluent: isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 35° C.; UV detection: 220 nm].

(231) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.47 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.81 (q, 1H), 5.02-5.13 (m, 2H), 6.88 (br. s, 1H), 7.33 (t, 1H), 7.30-7.35 (m, 1H), 7.48-7.56 (m, 2H), 7.59-7.65 (m, 3H), 7.72-7.78 (m, 2H).

Example 57

5-(4-Chlorophenyl)-2-({1-[3-(difluoromethoxy)phenyl]-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(232) Analytical chiral HPLC: R.sub.t=6.70 min, d.e.=100% [column: Daicel Chiralcel OX-H 5, 250×4.6 mm; eluent: isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 35° C.; UV detection: 220 nm].

(233) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.47 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.23-4.35 (m, 1H), 4.82 (quin, 1H), 5.07 (s, 2H), 5.75 (d, 1H), 6.90 (d, 1H), 7.33 (t, 1H), 7.30-7.35 (m, 1H), 7.48-7.56 (m, 2H), 7.59-7.65 (m, 3H), 7.72-7.78 (m, 2H).

Example 58

2-({1-(2-Chloro-5-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(234) ##STR00046##

(235) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added (2-chloro-5-fluorophenyl)boronic acid (322.31 mg, 1.85 mmol) and copper(II) acetate (335.75 mg, 1.85 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for 10 days. Over this time, extra boronic acid (322.31 mg in total, 1.85 mmol) was added portionwise in a daily fashion. The resulting reaction mixture was concentrated in vacuo, then diluted with ethyl acetate and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (61 mg, 0.11 mmol, 98% purity) was obtained as a mixture of diastereomers (yield 11.5%).

(236) LC/MS [method 2]: R.sub.t=3.02 min; MS [ESIpos]: m/z=561 (M+H).sup.+

(237) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.40 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.36 (m, 1H), 4.63-4.72 (m, 1H), 5.01-5.13 (m, 2H), 5.52 (br. s, 1H), 6.90 (dd, 1H), 7.52 (td, 1H), 7.60-7.69 (m, 3H), 7.73-7.79 (m, 3H).

(238) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 58 mg dissolved in 3 ml ethanol/isohexane (2:1); injection volume: 1 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 25° C.; UV detection: 220 nm]. After separation, 25 mg of diastereomer 1 (Example 59), which eluted first, and 25 mg of diastereomer 2 (Example 60), which eluted later, were isolated.

Example 59

2-{[1-(2-Chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(239) LC/MS [method 2]: R.sub.t=3.02 min; MS [ESIpos]: m/z=561 (M+H).sup.+

(240) Analytical chiral HPLC: R.sub.t=5.43 min, d.e.=100% [column: Daicel Chiralcel OX-H 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(241) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.40 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.35 (m, 1H), 4.67 (quin, 1H), 5.01-5.12 (m, 2H), 5.53 (d, 1H), 6.89 (d, 1H), 7.52 (td, 1H), 7.60-7.68 (m, 3H), 7.72-7.79 (m, 3H).

Example 60

2-{[1-(2-Chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(242) LC/MS [method 2]: R.sub.t=3.02 min; MS [ESIpos]: m/z=561 (M+H).sup.+

(243) Analytical chiral HPLC: R.sub.t=6.11 min, d.e.=100% [column: Daicel Chiralcel OX-H 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(244) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.40 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.67 (quin, 1H), 5.07 (s, 2H), 5.53 (d, 1H), 6.90 (d, 1H), 7.52 (td, 1H), 7.60-7.68 (m, 3H), 7.73-7.79 (m, 3H).

Example 61

5-(4-Chlorophenyl)-2-({1-(2,3-dichlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(245) ##STR00047##

(246) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (500 mg, 0.92 mmol, 80% purity) in pyridine (12 ml) were added (2,3-dichlorophenyl)boronic acid (176.36 mg, 0.92 mmol) and copper(II) acetate (335.75 mg, 1.85 mmol). The reaction mixture was heated to 60° C. for 1 h and then stirred at room temperature for 24 h, after which extra boronic acid (80 mg, 0.42 mmol) was added due to incomplete conversion. The reaction mixture was further stirred at room temperature for 5 days. Over this time, two additional portions of boronic acid (160 mg in total, 0.84 mmol) were added. After this, the resulting reaction mixture was concentrated in vacuo, then diluted with MTBE and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (148 mg, 0.25 mmol, 97.3% purity) was obtained as a mixture of diastereomers (yield 27%).

(247) LC/MS [method 2]: R.sub.t=3.19 min; MS [ESIpos]: m/z=577 (M+H).sup.+

(248) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.39 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.35 (m, 1H), 4.60-4.71 (m, 1H), 5.02-5.13 (m, 2H), 5.52 (br. s, 1H), 6.89 (dd, 1H), 7.55 (t, 1H), 7.59-7.66 (m, 3H), 7.73-7.78 (m, 2H), 7.87 (dd, 1H).

(249) The two diastereomers were separated by preparative chiral HPLC (SFC) [sample preparation: 141 mg dissolved in 18 ml methanol; injection volume: 0.3 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: carbon dioxide/methanol 70:30; flow rate: 80 ml/min; temperature: 40° C.; UV detection: 210 nm]. After separation, 58.5 mg of diastereomer 1 (Example 62), which eluted first, and 53 mg of diastereomer 2 (Example 63), which eluted later, were isolated.

Example 62

5-(4-Chlorophenyl)-2-{[1-(2,3-dichlorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(250) LC/MS [method 2]: R.sub.t=3.21 min; MS [ESIpos]: m/z=577 (M+H).sup.+; 95% purity

(251) Analytical chiral HPLC (SFC): R.sub.t=3.09 min, d.e.=100% [column: Daicel Chiralcel OX-3 250×4 mm; eluent: carbon dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV detection: 220 nm].

(252) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.39 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.36 (m, 1H), 4.65 (br. s, 1H), 5.01-5.13 (m, 2H), 5.52 (d, 1H), 6.89 (d, 1H), 7.55 (t, 1H), 7.59-7.66 (m, 3H), 7.72-7.78 (m, 2H), 7.87 (dd, 1H).

Example 63

5-(4-Chlorophenyl)-2-{[1-(2,3-dichlorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(253) LC/MS [method 2]: R.sub.t=3.20 min; MS [ESIpos]: m/z=577 (M+H).sup.+; 95% purity

(254) Analytical chiral HPLC (SFC): R.sub.t=3.38 min, d.e.=100% [column: Daicel Chiralcel OX-3 250×4 mm; eluent: carbon dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV detection: 220 nm].

(255) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.39 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.35 (m, 1H), 4.65 (br. s, 1H), 5.07 (s, 2H), 5.52 (d, 1H), 6.90 (d, 1H), 7.55 (t, 1H), 7.59-7.66 (m, 3H), 7.72-7.79 (m, 2H), 7.87 (dd, 1H).

Example 64

5-(4-Chlorophenyl)-2-({1-(2,3-difluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(256) ##STR00048##

(257) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (430 mg, 0.99 mmol) in pyridine (12.5 ml) were added (2,3-difluorophenyl)boronic acid (156.89 mg, 0.99 mmol) and copper(II) acetate (360.94 mg, 1.99 mmol). The reaction mixture was heated to 60° C. for 1 h and then stirred at room temperature for 24 h, after which extra boronic acid (80 mg, 0.51 mmol) was added due to incomplete conversion. The reaction mixture was further stirred at room temperature for five days. Over this time, five additional portions of boronic acid (400 mg in total, 2.54 mmol) were added. After this, the resulting reaction mixture was concentrated in vacuo, then diluted with MTBE and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (44 mg, 0.08 mmol) was obtained as a mixture of diastereomers (yield 8.1%).

(258) LC/MS [method 2]: R.sub.t=2.97 min; MS [ESIpos]: m/z=545 (M+H).sup.+

(259) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.42 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.76 (q, 1H), 5.02-5.13 (m, 2H), 6.89 (br. s, 1H), 7.35-7.43 (m, 1H), 7.45-7.51 (m, 1H), 7.59-7.71 (m, 3H), 7.72-7.79 (m, 2H).

(260) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 40 mg dissolved in 1 ml ethanol; injection volume: 0.5 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 35° C.; UV detection: 220 nm]. After separation, 18 mg of diastereomer 1 (Example 65), which eluted first, and 16 mg of diastereomer 2 (Example 66), which eluted later, were isolated.

Example 65

5-(4-Chlorophenyl)-2-{[1-(2,3-difluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(261) Analytical chiral HPLC: R.sub.t=5.74 min, d.e.=100% [column: Daicel Chiralcel OX-H 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 35° C.; UV detection: 220 nm].

(262) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.42 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.35 (m, 1H), 4.76 (quin, 1H), 5.02-5.13 (m, 2H), 5.58 (d, 1H), 6.89 (d, 1H), 7.35-7.44 (m, 1H), 7.45-7.52 (m, 1H), 7.59-7.72 (m, 3H), 7.72-7.78 (m, 2H).

Example 66

5-(4-Chlorophenyl)-2-{[1-(2,3-difluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(263) Analytical chiral HPLC: R.sub.t=6.59 min, d.e.=99.2% [column: Daicel Chiralcel OX-H 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 35° C.; UV detection: 220 nm].

(264) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.42 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.36 (m, 1H), 4.76 (quin, 1H), 5.07 (s, 2H), 5.58 (d, 1H), 6.90 (d, 1H), 7.35-7.43 (m, 1H), 7.44-7.51 (m, 1H), 7.59-7.72 (m, 3H), 7.72-7.78 (m, 2H).

Example 67

2-{[1-(2-Chloro-3-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(265) ##STR00049##

(266) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (500 mg, 0.92 mmol, 80% purity) in pyridine (12 ml) were added (2-chloro-3-fluorophenyl)boronic acid (161.15 mg, 0.92 mmol) and copper(II) acetate (335.75 mg, 1.85 mmol). The reaction mixture was heated to 60° C. for 1 h and then stirred at room temperature for 24 h, after which extra boronic acid (75 mg, 0.43 mmol) was added due to incomplete conversion. The reaction mixture was further stirred at room temperature for six days. Over this time, five additional portions of boronic acid (375 mg in total, 2.15 mmol) were added. After this, the resulting reaction mixture was concentrated in vacuo, then diluted with MTBE and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and 91 mg of the desired compound as a diastereomeric mixture still containing some impurities were isolated.

(267) A further purification by preparative chiral HPLC yielded the two pure, separated diastereomers [sample preparation: 90 mg dissolved in 3 ml ethanol; injection volume: 0.3 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 35° C.; UV detection: 220 nm]. After separation, 20 mg of diastereomer 1 (Example 67), which eluted first, and 21 mg of diastereomer 2 (Example 68), which eluted later, were isolated.

(268) LC/MS [method 2]: R.sub.t=3.01 min; MS [ESIpos]: m/z=561 (M+H).sup.+

(269) Analytical chiral HPLC: R.sub.t=6.22 min, d.e.=100% [column: Daicel Chiralcel OX-H 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 35° C.; UV detection: 220 nm].

(270) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.39 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.36 (m, 1H), 4.66 (quin, 1H), 5.01-5.14 (m, 2H), 5.53 (d, 1H), 6.89 (d, 1H), 7.48-7.70 (m, 5H), 7.72-7.78 (m, 2H).

Example 68

2-{[1-(2-Chloro-3-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(271) LC/MS [method 2]: R.sub.t=3.01 min; MS [ESIpos]: m/z=561 (M+H).sup.+

(272) Analytical chiral HPLC: R.sub.t=7.94 min, d.e.=100% [column: Daicel Chiralcel OX-H 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 35° C.; UV detection: 220 nm].

(273) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.40 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.36 (m, 1H), 4.66 (quin, 1H), 5.07 (s, 2H), 5.53 (d, 1H), 6.90 (d, 1H), 7.49-7.70 (m, 5H), 7.72-7.78 (m, 2H).

Example 69

5-(4-Chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1-(2-methylphenyl)-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(274) ##STR00050##

(275) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added (2-methylphenyl)boronic acid (251.32 mg, 1.85 mmol) and copper(II) acetate (335.75 mg, 1.85 mmol). The reaction mixture was stirred at room temperature for 5 days, after which extra boronic acid (62.8 mg, 0.46 mmol, 0.5 eq.) was added due to incomplete conversion. After stirring for two additional days, the reaction mixture was concentrated in vacuo, then diluted with MTBE and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (100 mg, 0.17 mmol) was obtained as a mixture of diastereomers (yield 17.2%, 90% purity).

(276) LC/MS [method 3]: R.sub.t=1.24 min; MS [ESIpos]: m/z=523 (M+H).sup.+

(277) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 98 mg dissolved in 2 ml ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 75:25; flow rate: 15 ml/min; temperature: 30° C.; UV detection: 220 nm]. After separation, 37 mg of diastereomer 1 (Example 70), which eluted first, and 39 mg of diastereomer 2 (Example 71), which eluted later, were isolated.

Example 70

5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(2-methylphenyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(278) LC/MS [method 3]: R.sub.t=1.24 min; MS [ESIpos]: m/z=523 (M+H).sup.+

(279) Analytical chiral HPLC: R.sub.t=7.65 min, d.e.=100% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(280) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.36 (d, 3H), 1.98 (s, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.35 (m, 1H), 4.54 (quin, 1H), 5.00-5.12 (m, 2H), 5.48 (d, 1H), 6.89 (d, 1H), 7.31-7.49 (m, 4H), 7.59-7.65 (m, 2H), 7.71-7.77 (m, 2H).

Example 71

5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(2-methylphenyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(281) Analytical chiral HPLC: R.sub.t=10.27 min, d.e.=100% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(282) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.37 (d, 3H), 1.98 (s, 3H), 3.84 (dd, 1H), 4.00 (dd, 1H), 4.24-4.35 (m, 1H), 4.54 (quin, 1H), 5.06 (s, 2H), 5.48 (d, 1H), 6.90 (d, 1H), 7.31-7.49 (m, 4H), 7.58-7.66 (m, 2H), 7.71-7.78 (m, 2H).

Example 72

5-(4-Chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1-[2-(trifluoromethyl)phenyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(283) ##STR00051##

(284) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (600 mg, 1.11 mmol, 80% purity) in pyridine (14.5 ml) were added [2-(trifluoromethyl)phenyl]boronic acid (421.30 mg, 2.22 mmol) and copper(II) acetate (402.9 mg, 2.22 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for 5 days, after which extra boronic acid (105 mg, 0.55 mmol, 0.5 eq.) was added due to incomplete conversion. After further stirring at room temperature overnight, the resulting reaction mixture was concentrated in vacuo, then diluted with MTBE and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (80 mg) was obtained as a mixture of diastereomers (yield 12.4%).

(285) LC/MS [method 2]: R.sub.t=3.19 min; MS [ESIpos]: m/z=577 (M+H).sup.+

(286) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 78 mg dissolved in 2 ml ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 75:25; flow rate: 15 ml/min; temperature: 30° C.; UV detection: 220 nm]. After separation, 34 mg of diastereomer 1 (Example 73), which eluted first, and 30 mg of diastereomer 2 (Example 74), which eluted later, were isolated.

Example 73

5-(4-Chlorophenyl)-2-({5-(1-hydroxyethyl)-1-[2-(trifluoromethyl)phenyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)

(287) Analytical chiral HPLC: R.sub.t=6.16 min, d.e.=100% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(288) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.36 (d, 3H), 3.84 (dd, 1H), 4.00 (dd, 1H), 4.24-4.35 (m, 1H), 4.57 (quin, 1H), 4.99-5.12 (m, 2H), 5.50 (d, 1H), 6.89 (d, 1H), 7.59-7.65 (m, 2H), 7.66-7.71 (m, 1H), 7.72-7.76 (m, 2H), 7.77-7.90 (m, 2H), 7.93-7.99 (m, 1H).

Example 74

5-(4-Chlorophenyl)-2-({5-(1-hydroxyethyl)-1-[2-(trifluoromethyl)phenyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)

(289) Analytical chiral HPLC: R.sub.t=8.67 min, d.e.=100% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(290) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.36 (d, 3H), 3.84 (dd, 1H), 3.99 (dd, 1H), 4.24-4.35 (m, 1H), 4.54-4.62 (m, 1H), 5.05 (s, 2H), 5.50 (d, 1H), 6.90 (d, 1H), 7.60-7.65 (m, 2H), 7.67-7.71 (m, 1H), 7.72-7.90 (m, 4H), 7.93-7.98 (m, 1H).

Example 75

5-(4-Chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(291) ##STR00052##

(292) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (430 mg, 0.795 mmol, 80% purity) in pyridine (10 ml) were added (3-fluorophenyl)boronic acid (222.432 mg, 1.59 mmol) and copper(II) acetate (288.75 mg, 1.59 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for 5 days, after which extra boronic acid (55.6 mg, 0.40 mmol) was added due to incomplete conversion. The reaction mixture was again heated to 60° C. for 2 h, followed by stirring at room temperature overnight. The resulting reaction mixture was concentrated in vacuo, then diluted with MTBE and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (100 mg, 0.19 mmol) was obtained as a mixture of diastereomers (yield 23.9%).

(293) LC/MS [method 2]: R.sub.t=2.99 min; MS [ESIpos]: m/z=527 (M+H).sup.+

(294) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.47 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.83 (q, 1H), 5.02-5.13 (m, 2H), 6.89 (br. s, 1H), 7.38 (td, 1H), 7.48-7.66 (m, 5H), 7.72-7.78 (m, 2H).

(295) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 97 mg dissolved in 4 ml ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 30° C.; UV detection: 220 nm]. After separation, 36 mg of (1S)-diastereomer (Example 76), which eluted first, and 40 mg of (1R)-diastereomer (Example 77), which eluted later, were isolated.

Example 76

5-(4-Chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(296) ##STR00053##

(297) LC/MS [method 3]: R.sub.t=1.24 min; MS [ESIpos]: m/z=527 (M+H).sup.+

(298) Analytical chiral HPLC: R.sub.t=9.71 min, d.e.=100% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(299) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.47 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.37 (m, 1H), 4.82 (quin, 1H), 5.01-5.13 (m, 2H), 5.76 (d, 1H), 6.89 (d, 1H), 7.38 (td, 1H), 7.48-7.66 (m, 5H), 7.72-7.79 (m, 2H).

(300) The absolute stereochemistry of the compound was determined by additionally performing the same reaction with enantiopure diastereomer 5-(4-chlorophenyl)-2-({5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (Example 5A) as the starting material and comparison of the two respective products by analytical chiral HPLC.

Example 77

5-(4-Chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1R)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(301) ##STR00054##

(302) LC/MS [method 2]: R.sub.t=2.93 min; MS [ESIpos]: m/z=527 (M+H).sup.+

(303) Analytical chiral HPLC: R.sub.t=13.60 min, d.e.=100% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(304) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.47 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.36 (m, 1H), 4.83 (quin, 1H), 5.07 (s, 2H), 5.76 (d, 1H), 6.90 (d, 1H), 7.38 (td, 1H), 7.48-7.65 (m, 5H), 7.72-7.78 (m, 2H).

(305) The absolute stereochemistry of the compound was determined by additionally performing the same reaction with enantiopure diastereomer 5-(4-chlorophenyl)-2-({5-[(1R)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (Example 6A) as the starting material and comparison of the two respective products by analytical chiral HPLC.

Example 78

5-(4-Chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(306) ##STR00055##

(307) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (430 mg, 0.795 mmol, 80% purity) in pyridine (10 ml) were added (3-chlorophenyl)boronic acid (248.59 mg, 1.59 mmol) and copper(II) acetate (288.75 mg, 1.59 mmol). The reaction mixture was heated to 60° C. for 2 h and then stirred at room temperature for 5 days, after which extra boronic acid (62.1 mg, 0.40 mmol) was added due to incomplete conversion. The reaction mixture was again heated to 60° C. for 2 h, followed by stirring at room temperature overnight. The resulting reaction mixture was concentrated in vacuo, then diluted with MTBE and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (130 mg, 0.24 mmol) was obtained as a mixture of diastereomers (yield 30.1%).

(308) LC/MS [method 2]: R.sub.t=3.19 min; MS [ESIpos]: m/z=543 (M+H).sup.+

(309) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.47 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.81 (q, 1H), 5.02-5.13 (m, 2H), 6.89 (br. s, 1H), 7.56-7.67 (m, 5H), 7.72-7.79 (m, 3H).

(310) The two diastereomers were separated by preparative chiral HPLC [sample preparation: 128 mg dissolved in 4 ml ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 30° C.; UV detection: 220 nm]. After separation, 52 mg of (1S)-diastereomer (Example 79), which eluted first, and 49 mg of (1R)-diastereomer (Example 80), which eluted later, were isolated.

Example 79

5-(4-Chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(311) ##STR00056##

(312) LC/MS [method 2]: R.sub.t=3.14 min; MS [ESIpos]: m/z=543 (M+H).sup.+

(313) Analytical chiral HPLC: R.sub.t=9.96 min, d.e.=100% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 35° C.; UV detection: 220 nm].

(314) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.47 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.23-4.36 (m, 1H), 4.81 (quin, 1H), 5.01-5.13 (m, 2H), 5.76 (d, 1H), 6.89 (d, 1H), 7.56-7.66 (m, 5H), 7.71-7.79 (m, 3H).

(315) .sup.13C NMR (125 MHz, DMSO-d.sub.6): δ [ppm] 21.3, 42.1, 42.2, 59.6, 65.5, 123.0, 124.5, 124.6, 125.3, 128.5, 128.9 (2×), 130.0 (2×), 130.7, 133.0, 135.2, 138.2, 144.8, 153.1, 157.8, 158.6.

(316) The absolute stereochemistry of the compound was determined by additionally performing the same reaction with enantiopure diastereomer 5-(4-chlorophenyl)-2-({5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (Example 5A) as the starting material and comparison of the two respective products by analytical chiral HPLC.

Example 80

5-(4-Chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1R)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(317) ##STR00057##

(318) LC/MS [method 2]: R.sub.t=3.15 min; MS [ESIpos]: m/z=543 (M+H).sup.+

(319) Analytical chiral HPLC: R.sub.t=14.41 min, d.e.=100% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 35° C.; UV detection: 220 nm].

(320) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.47 (d, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.37 (m, 1H), 4.81 (quin, 1H), 5.07 (s, 2H), 5.76 (d, 1H), 6.90 (d, 1H), 7.56-7.66 (m, 5H), 7.71-7.79 (m, 3H).

(321) The absolute stereochemistry of the compound was determined by additionally performing the same reaction with enantiopure diastereomer 5-(4-chlorophenyl)-2-({5-[(1R)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (Example 6A) as the starting material and comparison of the two respective products by analytical chiral HPLC.

Example 81

5-(4-Chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)

(322) ##STR00058##

(323) To a solution of 5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (2.10 g, 3.88 mmol, 80% purity) in pyridine (50 ml) were added (2-chlorophenyl)boronic acid (1.214 g, 7.76 mmol) and copper(II) acetate (1.410 g, 7.76 mmol). The reaction mixture was heated to 60° C. for 1 h and then stirred at room temperature for 5 days, after which extra boronic acid (303 mg, 1.94 mmol) was added due to incomplete conversion. After stirring at room temperature for two additional days, the resulting reaction mixture was concentrated in vacuo, then diluted with MTBE and quenched with aqueous hydrochloric acid (0.5 M). After phase separation, the aqueous phase was extracted twice with MTBE. The combined organic phases were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC [method 4], and the desired compound (580 mg, 1.01 mmol, 95% purity) was obtained as a mixture of diastereomers (yield 26.1%).

(324) LC/MS [method 3]: R.sub.t=1.24 min; MS [ESIpos]: m/z=543 (M+H).sup.+

(325) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.38 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.55-4.64 (m, 1H), 5.01-5.13 (m, 2H), 6.85-6.94 (m, 1H), 7.50-7.65 (m, 5H), 7.67-7.78 (m, 3H).

(326) The two diastereomers were separated by preparative chiral HPLC (SFC) [sample preparation: 575 mg dissolved in 35 ml methanol; injection volume: 0.4 ml; column: Daicel Chiralcel® OX-H 5 μm, 250×20 mm; eluent: carbon dioxide/methanol 70:30; flow rate: 80 ml/min; temperature: 40° C.; UV detection: 210 nm]. After separation, 206 mg of (1S)-diastereomer (Example 82), which eluted first, and 189 mg of (1R)-diastereomer (Example 83), which eluted later, were isolated.

Example 82

5-(4-Chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(327) ##STR00059##

(328) LC/MS [method 3]: R.sub.t=1.24 min; MS [ESIpos]: m/z=543 (M+H).sup.+

(329) Analytical chiral HPLC: R.sub.t=8.34 min, d.e.=100% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(330) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.38 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.59 (q, 1H), 5.01-5.13 (m, 2H), 5.50 (br. s, 1H), 6.90 (d, 1H), 7.50-7.65 (m, 5H), 7.67-7.78 (m, 3H).

(331) The absolute stereochemistry of the compound was determined by additionally performing the same reaction with enantiopure diastereomer 5-(4-chlorophenyl)-2-({5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (Example 5A) as the starting material and comparison of the two respective products by analytical chiral HPLC.

Example 83

5-(4-Chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1R)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

(332) ##STR00060##

(333) Analytical chiral HPLC: R.sub.t=11.88 min, d.e.=98.1% [column: LUX Cellulose-4, 5 μm, 250×4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40° C.; UV detection: 220 nm].

(334) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm] 1.38 (d, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.36 (m, 1H), 4.54-4.65 (m, 1H), 5.07 (s, 2H), 5.51 (br. s, 1H), 6.90 (d, 1H), 7.50-7.65 (m, 5H), 7.68-7.79 (m, 3H).

(335) The absolute stereochemistry of the compound was determined by additionally performing the same reaction with enantiopure diastereomer 5-(4-chlorophenyl)-2-({5-[(1R)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one (Example 6A) as the starting material and comparison of the two respective products by analytical chiral HPLC.

B. EVALUATION OF BIOLOGICAL ACTIVITY

(336) Abbreviations and Acronyms:

(337) Acc. No. accession number AVP arginine vasopressin B.sub.max maximal ligand binding capacity BSA bovine serum albumin cAMP cyclic adenosine monophosphate Cat. No. catalogue number cDNA complementary deoxyribonucleic acid CHO chinese hamster ovary CRE cAMP response element Ct cycle threshold DMEM/F12 Dulbecco's modified Eagle's medium/Ham's F12 medium (1:1) DNA deoxyribonucleic acid DTT dithiothreitol EC.sub.50 half-maximal effective concentration EDTA ethylenediamine-tetraacetic acid FAM carboxyfluorescein succinimidyl ester f.c. final concentration FCS fetal calf serum HEPES 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid IC.sub.50 half-maximal inhibitory concentration K.sub.d dissociation constant K.sub.i dissociation constant of an inhibitor mRNA messenger ribonucleic acid PBS phosphate buffered saline p.o. per os, peroral RNA ribonucleic acid RTPCR real-time polymerase chain reaction SPA scintillation proximity assay TAMRA carboxytetramethylrhodamine TRIS 2-amino-2-hydroxymethylpropane-1,3-diol

(338) Demonstration of the activity of the compounds of the present invention may be accomplished through in vitro, ex vivo, and in vivo assays that are well known in the art. For example, to demonstrate the activity of the compounds of the present invention, the following assays may be used.

(339) B-1. Cellular in Vitro Assay for Determining Vasopressin Receptor Activity

(340) The identification of agonists and antagonists of the V1a and V2 vasopressin receptors from humans, rats and dogs as well as the quantification of the activity of the compounds of the invention is carried out using recombinant cell lines. These cell lines originally derive from a hamster's ovary epithelial cell (Chinese Hamster Ovary, CHO K1, ATCC: American Type Culture Collection, Manassas, Va. 20108, USA). The test cell lines constitutively express the human, rat or dog V1a or V2 receptors. In case of the G.sub.αy-coupled V1a receptors, cells are also stably transfected with a modified form of the calcium-sensitive photoproteins aequorin (human and rat V1a) or obelin (dog V1a), which, after reconstitution with the cofactor coelenterazine, emit light when there are increases in free calcium concentrations [Rizzuto R, Simpson A W, Brini M, Pozzan T, Nature 358, 325-327 (1992); Illarionov B A, Bondar V S, Illarionova V A, Vysotski E S, Gene 153 (2), 273-274 (1995)]. The resulting vasopressin receptor cells react to stimulation of the recombinantly expressed V1a receptors by intracellular release of calcium ions, which can be quantified by the resulting photoprotein luminescence. The G.sub.s-coupled V2 receptors are stably transfected into cell lines expressing the gene for firefly luciferase under control of a CRE-responsible promoter. Activation of V2 receptors induces the activation of the CRE-responsive promoter via cAMP increase, thereby inducing the expression of firefly luciferase. The light emitted by photoproteins of V1a cell lines as well as the light emitted by firefly luciferase of V2 cell lines corresponds to the activation or inhibition of the respective vasopressin receptor. The bioluminescence of the cell lines is detected using a suitable luminometer [Milligan G, Marshall F, Rees S, Trends in Pharmacological Sciences 17, 235-237 (1996)].

(341) Test Procedure:

(342) Vasopressin V1a Receptor Cell Lines:

(343) On the day before the assay, the cells are plated out in culture medium (DMEM/F12, 2% FCS, 2 mM glutamine, 10 mM HEPES, 5 μg/ml coelenterazine) in 384-well microtiter plates and kept in a cell incubator (96% humidity, 5% v/v CO.sub.2, 37° C.). On the day of the assay, test compounds in various concentrations are placed for 10 minutes in the wells of the microtiter plate before the agonist [Arg.sup.8]-vasopressin at EC.sub.50 concentration is added. The resulting light signal is measured immediately in the luminometer.

(344) Vasopressin V2 Receptor Cell Lines:

(345) On the day before the assay, the cells are plated out in culture medium (DMEM/F12, 2% FCS, 2 mM glutamine, 10 mM HEPES) in 384-well microtiter plates and kept in a cell incubator (96% humidity, 5% v/v CO2, 37° C.). On the day of the assay, test compounds in various concentrations and the agonist [Arg.sup.8]-vasopressin at EC.sub.50 concentration are added together to the wells, and plates are incubated for 3 hours in a cell incubator. Upon addition of the cell lysis reagent Triton and the substrate luciferin, luminescence of firefly luciferase is measured in a luminometer.

(346) Table 1A below lists individual IC.sub.50 values for the compounds of the invention (including diastereomeric mixtures as well as separated, enantiopure diastereomers) that were obtained from cell lines transfected with the human V1a or V2 receptor:

(347) TABLE-US-00001 TABLE 1A Example IC.sub.50 hV1a IC.sub.50 hV2 No. [μM] [μM] 1 0.0060 0.0025 2 0.0050 0.0087 3 0.0010 0.0056 4 0.0004 0.0053 5 0.0004 0.0018 6 0.0106 0.0017 7 0.0076 0.0026 8 0.0012 0.0107 9 0.0004 0.0023 10 0.0014 0.0014 11 0.0004 0.0003 12 0.0013 0.0011 13 0.0062 0.0014 14 0.0013 0.0004 15 0.0384 0.0041 16 0.0031 0.0060 17 0.0027 0.0034 18 0.0141 0.0086 19 0.0124 0.0014 20 0.0038 0.0008 21 0.0578 0.0022 22 0.0244 0.0023 23 0.0122 0.0009 24 0.1200 0.0020 25 0.0072 0.0036 26 0.0031 0.0030 27 0.0437 0.0077 28 0.0013 0.0002 29 0.0029 0.0002 30 0.0716 0.0004 31 0.0016 0.0009 32 0.0009 0.0010 33 0.0016 0.0023 34 0.0004 0.0012 35 0.0005 0.0016 36 0.0008 0.0028 37 0.0005 0.0007 38 0.0006 0.0009 39 0.0015 0.0035 40 0.0015 0.0072 41 0.0018 0.0079 42 0.0051 0.0127 43 0.0062 0.0012 44 0.0061 0.0012 45 0.0921 0.0033 46 0.0063 0.0021 47 0.0189 0.0037 48 0.0032 0.0024 49 0.0018 0.0126 50 0.0013 0.0100 51 0.0030 0.0223 52 0.0039 0.0004 53 0.0079 0.0018 54 0.0397 0.0016 55 0.0148 0.0042 56 0.0024 0.0014 57 0.0382 0.0082 58 0.0002 0.0015 59 0.0005 0.0024 60 0.0005 0.0052 61 0.0032 0.0002 62 0.0078 0.0009 63 0.0516 0.0019 64 0.0081 0.0051 65 0.0025 0.0033 66 0.0040 0.0019 67 0.0021 0.0027 68 0.0033 0.0013 70 0.0005 0.0011 71 0.0006 0.0021 73 0.0011 0.0070 74 0.0022 0.0247 75 0.0029 0.0066 76 0.0025 0.0051 77 0.0125 0.0135 78 0.0104 0.0031 79 0.0036 0.0017 80 0.0463 0.0051 82 0.0007 0.0023 83 0.0010 0.0067

(348) The IC.sub.50 data listed in Table 1A demonstrate that the compounds of the present invention are acting as highly potent dual antagonists of vasopressin V1a and V2 receptors.

(349) For comparative purposes, selected phenyl-triazole and imidazole derivatives that were regarded to be representative of closest prior art (cf. Int. Pat. Appl. WO 2011/104322-A1 and example compounds described therein) were also tested in the cellular V1a and V2 assays described above. IC.sub.50 values for these compounds obtained from cell lines transfected with the human V1a or V2 receptor are listed in Table 1B below:

(350) TABLE-US-00002 TABLE 1B Example No. IC.sub.50 hV1a IC.sub.50 hV2 WO 2011/104322 [μM] [μM] 54 0.0166 0.0564 56 0.0013 0.0067 60 0.0542 0.0326 68 0.0060 0.0083 101 0.0422 0.0238 110 0.0152 0.0043

(351) B-2. Radioactive Binding Assay

(352) IC.sub.50 and K, values were determined in a radioactive binding competition SPA assay using membrane fractions of recombinant CHO cell lines expressing the respective human, rat or dog vasopressin V1a and V2 receptors. These cells derive from a hamster's ovary epithelial cell (Chinese Hamster Ovary, CHO K1, ATCC: American Type Culture Collection, Manassas, Va. 20108, USA). In addition, the cells are stably transfected with the human, rat or dog V1a or V2 receptor. The membrane preparations were subjected to the radioactive receptor binding competition assay described below.

(353) The respective vasopressin receptor-transfected CHO cells were grown in an appropriate quantity in T-175 flasks with DMEM/F12, 10% FCS, 15 mM HEPES, 1 mg/ml G418 and kept in a cell incubator (96% humidity, 5% v/v CO2, 37° C.). After reaching the appropriate confluency, cells were harvested for membrane preparation. Cells were scraped into PBS and pelleted by gentle centrifugation at 200×g for 5 min at room temperature. Pellets were re-suspended in PBS and again centrifugated. After repeating this step once more, the resulting pellets were shock-frozen at −80° C. for 30 min. Frozen pellets were re-suspended in ice-cold preparation buffer (50 mM TRIS, 2 mM EDTA, 2 mM DTT, cOmplete Protease Inhibitor Cocktail) and homogenized at 2000 rpm for 35 seconds (Polytron PT3000, Kinematica). The homogenate was cooled down for 2 min on ice, and the homogenization was repeated twice. The resulting homogenate was centrifugated at 500×g for 10 min at 4° C. Membranes were pelleted at 4500×g for 20 min at 4° C., re-suspended in storing buffer (7.5 mM TRIS, 12.5 mM MgCl.sub.2, 0.3 mM EDTA, 250 mM sucrose, cOmplete Protease Inhibitor Cocktail) and homogenized at 2000 rpm for 2 seconds (Polytron PT3000, Kinematica). The protein concentration was determined by using the BCA Protein Assay (Thermo Scientific Pierce), and the membrane preparations were stored at −80° C. On the day of use, aliquots were thawed and briefly vortexed.

(354) For the determination of the receptor binding affinity of test compounds, an SPA assay was set-up as follows. For each membrane preparation, K.sub.d and B.sub.max values were determined. From these data, the number of SPA beads (WGA PVT beads, PerkinElmer, 200 μg/well), the concentration of radioactive ligand (.sup.3H-AVP, PerkinElmer, 2.431 TBq/mmol, f.c. 1-2×K.sub.d) and the amount of the respective membrane preparation (10 μg protein/well) were matched to the assay volume (100 μl) in binding buffer (50 mM TRIS, 0.2% BSA) in a 96-well plate. The test compounds were diluted in binding buffer (f.c. 10.sup.−4 M to 10.sup.−12 M) and subjected to the assay. Plates were gently shaken for 1-3 hours at room temperature and further incubated for 1-2 hours. Signals generated by bound .sup.3H-AVP were measured using a β-counter (1450 Microbeta Trilux). From these results, IC.sub.50 and K, values for the tested compounds were calculated using GraphPad Prism.

(355) B-3. Cellular in Vitro Assay for Detecting the Action of Vasopressin V1a Receptor Antagonists on the Regulation of Pro-Fibrotic Genes

(356) The cell line H9C2 (American Type Culture Collection No. CRL-1446), described as a cardiomyocyte type isolated from rat cardiac tissue, endogenously expresses the vasopressin V1A receptor AVPR1A in high copy number, whereas AVPR2 expression cannot be detected. For cell assays for the inhibition of AVPR1A receptor-dependent regulation of gene expression by receptor antagonists, the procedure is as follows:

(357) H9C2 cells are seeded in 6-well microtiter plates for cell culture at a cell density of 50 000 cells/well in 2.0 ml of Opti-MEM medium (Invitrogen Corp., Carlsbad, Calif., USA, Cat. No. 11058-021) and held in a cell incubator (96% humidity, 8% v/v CO2, 37° C.). After 24 hours, sets of three wells (triplicate) are charged with vehicle solution (negative control) and vasopressin solution ([Arg.sup.8]-vasopressin acetate, Sigma, Cat. No. V9879), or test compound (dissolved in vehicle: water with 20% v/v ethanol) and vasopressin solution. In the cell culture, the final vasopressin concentration is 1 nM. The test compound solution is added to the cell culture in small volumes, so that a final concentration of 0.03% of ethanol in the cell assay is not exceeded. After an incubation time of 5 hours, the culture supernatant is drawn off under suction, the adherent cells are lysed in 350 μl of RLT buffer (Qiagen, Cat. No. 79216), and the RNA is isolated from the lysate using the RNeasy kit (Qiagen, Cat. No. 74104). This is followed by DNAse digestion (Invitrogen, Cat. No. 18068-015), cDNA synthesis (Promaga, ImProm-II Reverse Transcription System, Cat. No. A3800) and RTPCR (pPCR MasterMix RT-QP2X-03-075, Eurogentec, Seraing, Belgium). All procedures take place in accordance with the working protocols of the test reagents' manufacturers. The primer sets for the RTPCR are selected on the basis of the mRNA gene sequences (NCBI GenBank Entrez Nucleotide Data Base) using the Primer3Plus program with 6-FAM TAMRA-labelled probes. The RTPCR for determining the relative mRNA expression in the cells of the various assay batches is carried out using the Applied Biosystems ABI Prism 7700 Sequence Detector in 384-well microtiter plate format in accordance with the instrument operating instructions. The relative gene expression is represented by the delta-delta Ct value [Applied Biosystems, User Bulletin No. 2 ABI Prism 7700 SDS, Dec. 11, 1997 (updated 10/2001)] with reference to the level of expression of the ribosomal protein L-32 gene (GenBank Acc. No. NM_013226) and the threshold Ct value of Ct=35.

(358) B-4. In Vivo Assay for Detecting Cardiovascular Effects: Blood Pressure Measurement in Anaesthetized Rats (Vasopressin ‘Challenge’ model)

(359) In male Sprague-Dawley rats (250-350 g body weight) under ketamine/xylazine/pentobarbital injection anaesthesia, polyethylene tubes (PE-50, Intramedic®), which are prefilled with heparin-containing (500 IU/ml) isotonic sodium chloride solution, are introduced into the jugular vein and the femoral vein and then tied in. Via one venous access, with the aid of a syringe, Arg-vasopressin is injected; the test substance is administered via the second venous access. For determination of the systolic blood pressure, a pressure catheter (Millar SPR-320 2F) is tied into the carotid artery. The arterial catheter is connected to a pressure transducer which feeds its signals to a recording computer equipped with suitable recording software. In a typical experiment, the experimental animal is administered 3-4 successive bolus injections at intervals of 10-15 min with a defined amount of Arg-vasopressin (30 ng/kg) in isotonic sodium chloride solution. When the blood pressure has reached initial levels again, the test substance is administered as a bolus, with subsequent continuous infusion, in a suitable solvent. After this, at defined intervals (10-15 min), the same amount of Arg-vasopressin as at the start is administered again. On the basis of the blood pressure values, a determination is made of the extent to which the test substance counteracts the hypertensive effect of Arg-vasopressin. Control animals only receive solvent instead of the test substance.

(360) Following intravenous administration, the compounds of the invention, in comparison to the solvent controls, bring about an inhibition of the blood pressure increase caused by Arg-vasopressin.

(361) B-5. In Vivo Assay for Detecting Cardiovascular Effects: Diuresis Investigations in Conscious Rats Kept in Metabolism Cages

(362) Wistar rats (220-450 g body weight) are kept with free access to feed (Altromin) and drinking water. During the experiment, the animals are kept with free access to drinking water for 4 to 8 or up to 24 hours individually in metabolism cages suitable for rats of this weight class (Tecniplast Deutschland GmbH, D-82383 HohenpeiBenberg). At the beginning of the experiment, the animals are administered the test substance in a volume of 1 to 3 ml/kg body weight of a suitable solvent by means of gavage into the stomach. Control animals only receive solvent. Controls and substance tests are carried out in parallel on the same day. Control groups and substance-dose groups each consist of 4 to 8 animals. During the experiment, the urine excreted by the animals is collected continuously in a receiver at the base of the cage. The volume of urine per time unit is determined separately for each animal, and the concentration of urinary electrolytes is measured by standard methods of flame photometry. Before the beginning of the experiment, the body weight of the individual animals is determined.

(363) Following oral administration, in comparison with the solvent control applications, the compounds of the invention bring about an increased excretion of urine, which is based essentially on an increased excretion of water (aquaresis).

(364) Table 2A below shows observed changes in urinary excretion relative to solvent control (=100%) for exemplary compounds of the invention at two different dosages:

(365) TABLE-US-00003 TABLE 2A Urinary volume Urinary volume Dosage [% vs. Dosage [% vs. Example p.o. control = p.o. control = No. [mg/kg] 100%] [mg/kg] 100%] 1 0.3 194 3.0 588 2 0.3 194 3.0 588 3 0.3 89 3.0 450 4 0.3 91 1.0 190 5 0.3 166 1.0 438 6 0.3 132 1.0 439 11 0.3 159 3.0 443 12 0.3 96 3.0 323 14 — — 1.0 753 15 0.3 412 3.0 1085 17 0.3 404 1.0 819 29 0.3 404 1.0 983 70 0.3 139 3.0 595 76 0.3 350 3.0 1257 79 0.3 612 3.0 1312 82 0.3 220 3.0 828 83 0.3 279 3.0 1094

(366) For comparative purposes, selected phenyl-triazole and imidazole derivatives that were regarded to be representative of closest prior art (cf. Int. Pat. Appl. WO 2011/104322-A1 and example compounds described therein) were also tested for diuretic effect in this assay. Observed changes in urinary excretion relative to solvent control (=100%) at two different dosages are shown in Table 2B below:

(367) TABLE-US-00004 TABLE 2B Urinary volume Urinary volume Dosage [% vs. Dosage [% vs. Example No. p.o. control = p.o. control = WO 2011/104322 [mg/kg] 100%] [mg/kg] 100%] 54 0.3 85 3.0 188 56 0.3 128 3.0 85 60 0.3 96 3.0 84 68 0.3 87 3.0 121 101 0.3 111 3.0 255 110 0.3 114 3.0 274

(368) The results shown in Table 2A and 2B demonstrate that the compounds of the present invention are significantly more potent in vivo: Tested examples of the present invention gave rise to more than a threefold, in some cases to more than a tenfold increase in urinary volume versus the vehicle control group at a p.o. dose of 3 mg/kg, and most examples exhibited substantial aquaretic activity already at p.o. doses of 0.3 mg/kg or 1 mg/kg. This is in contrast to the phenyl-triazole and imidazole derivatives regarded to be representative of closest prior art which were not active at p.o. doses below 3 mg/kg and slightly active at 3 mg/kg.

(369) B-6. In Vivo Assay for Detecting Cardiovascular Effects: Hemodynamic Investigations in Anaesthetized Dogs

(370) Male beagle dogs (Beagle, Marshall BioResources) with a weight of between 10 and 15 kg are anaesthetized with pentobarbital (30 mg/kg i.v., Narcoren®, Merial, Germany) for the surgical interventions and hemodynamic and functional examinations. Pancuronium bromide (2 mg/animal i.v., Ratiopharm, Germany) serves additionally as a muscle relaxant. The dogs are intubated and ventilated with an oxygen/ambient air mixture (40/60%, about 3-4 L/min) Ventilation is carried out using a ventilator from GE Healthcare (Avance) and is monitored using an analyzer (Datex-Ohmeda, GE). Anaesthesia is maintained by continuous infusion of pentobarbital (50 μg/kg/min); fentanyl is used as an analgesic (10-40 μg/kg/h). An alternative to pentobarbital is to use isoflurane (1-2% by volume).

(371) In preparatory interventions, the dogs are fitted with a cardiac pacemaker. At a time of 21 days before the first drug testing (i.e. start of experiment), a cardiac pacemaker from Biotronik (Logos®) is implanted into a subcutaneous skin pocket and is contacted with the heart via a pacemaker electrode which is advanced through the external jugular vein, with transillumination, into the right ventricle. Thereafter all of the accesses are removed, and the dog wakes spontaneously from the anaesthesia. After a further 7 days (i.e. 14 days before the first drug testing), the above-described pacemaker is activated, and the heart is stimulated at a frequency of 220 beats per minute.

(372) The actual substance testing experiments are carried out 14 and 28 days after the beginning of pacemaker stimulation, using the following instrumentation: introduction of a bladder catheter for bladder relief and for measuring the flow of urine; attachment of ECG leads to the extremities for ECG measurement; introduction of a Fluidmedic PE 300 tube filled with sodium chloride solution into the femoral artery; this tube is connected to a pressure sensor (Braun Melsungen, Germany) for measuring systemic blood pressure; introduction of a Millar Tip catheter (type 350 PC, Millar Instruments, Houston, USA) through the left atrium or through a port secured in the carotid artery, for measuring cardiac hemodynamics; introduction of a Swan-Ganz catheter (CCOmbo 7.5F, Edwards, Irvine, USA) via the jugular vein into the pulmonary artery, for measuring cardiac output, oxygen saturation, pulmonary arterial pressures and central venous pressure; siting of a venous catheter in the cephalic vein, for infusing pentobarbital, for liquid replacement and for blood sampling (determination of plasma levels of the test substance or of other clinical blood values); siting of a venous catheter in the saphenous vein, for infusing fentanyl and for administration of the test substance; continuous infusion of vasopressin (Sigma, 4 mU/kg/min); test compounds are then administered and evaluated at different dosages under this vasopressin infusion.

(373) The primary signals are amplified if necessary (ACQ 7700 amplifier, DataSciences Inc., Minneapolis, USA, or Edwards-Vigilance-Monitor, Edwards, Irvine, USA) and subsequently fed into the Ponemah system (DataSciences Inc., Minneapolis, USA) for evaluation. The signals are recorded continuously throughout the experimental period, and are further processed digitally by the software and averaged over 30 seconds.

(374) Although the invention has been disclosed with reference to specific embodiments, it is apparent that other embodiments and variations of the invention may be devised by others skilled in the art without departing from the true spirit and scope of the invention. The claims are intended to be construed to include all such embodiments and equivalent variations.

(375) C. Examples Relating To Pharmaceutical Compositions

(376) Pharmaceutical compositions according to the present invention can be illustrated as follows:

(377) Sterile i.v. Solution:

(378) A 5 mg/mL solution of the desired compound of the invention can be made using sterile, injectable water, and the pH is adjusted if necessary. The solution is diluted for administration to 1-2 mg/mL with sterile 5% dextrose and is administered as an i.v. infusion over about 60 minutes.

(379) Lyophilized Powder for i.v Administration:

(380) A sterile preparation can be prepared with (i) 100-1000 mg of the desired compound of the invention as a lyophilized powder, (ii) 32-327 mg/mL sodium citrate, and (iii) 300-3000 mg Dextran 40. The formulation is reconstituted with sterile, injectable saline or 5% dextrose to a concentration of 10 to 20 mg/mL, which is further diluted with saline or 5% dextrose to 0.2 to 0.4 mg/mL, and is administered either as i.v. bolus or by i.v. infusion over 15-60 minutes.

(381) Intramuscular Suspension:

(382) The following solution or suspension can be prepared for intramuscular injection:

(383) 50 mg/mL of the desired, water-insoluble compound of the invention; 5 mg/mL sodium carboxymethylcellulose; 4 mg/mL Tween 80; 9 mg/mL sodium chloride; 9 mg/mL benzyl alcohol.

(384) Hard Shell Capsules:

(385) A large number of unit capsules are prepared by filling standard two-piece hard gelatin capsules each with 100 mg of the desired, powdered compound of the invention, 150 mg of lactose, 50 mg of cellulose and 6 mg of magnesium stearate.

(386) Soft Gelatin Capsules:

(387) A mixture of the desired compound of the invention in a digestible oil, such as soybean oil, cottonseed oil or olive oil, is prepared and injected by means of a positive displacement pump into molten gelatin to form soft gelatin capsules containing 100 mg of the active ingredient. The capsules are washed and dried. The desired compound of the invention can be dissolved in a mixture of polyethylene glycol, glycerin and sorbitol to prepare a water-miscible medicine mix.

(388) Tablets:

(389) A large number of tablets are prepared by conventional procedures so that the dosage unit is 100 mg of the desired compound of the invention, 0.2 mg of colloidal silicon dioxide, 5 mg of magnesium stearate, 275 mg of microcrystalline cellulose, 11 mg of starch, and 98.8 mg of lactose. Appropriate aqueous and non-aqueous coatings may be applied to increase palatability, improve elegance and stability, or delay absorption.