Substituted pyrazoles as JAK inhibitors

Abstract

A compound having the structure of below Formula (I), or pharmaceutically acceptable salts thereof, are useful as JAK inhibitors, ##STR00001##
wherein R.sub.1, R.sub.2, L.sub.1, L.sub.2, T and X are as herein described.

Claims

1. A compound of Formula (I): ##STR00061## or a pharmaceutically acceptable salt thereof, wherein: ring A is 5-6 membered heteroaryl; L.sub.1 and L.sub.2 are independently selected from a single bond, S(O).sub.2, S(O), C(O) or NHC(O); R.sub.1 is selected from H, C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, C.sub.3-7 cycloalkyl, 3-7 membered heterocycloalkyl, 5-6 membered aryl or 5-6 membered heteroaryl, wherein the C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, C.sub.3-7 cycloalkyl, 3-7 membered heterocycloalkyl, 5-6 membered aryl or 5-6 membered heteroaryl is optionally substituted with 1, 2, 3 or 4 R; or R.sub.1 is Formula (II): ##STR00062## wherein Formula (II) is optionally substituted with 1, 2, 3 or 4 R; R.sub.2 is selected from H, NH.sub.2, C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, C.sub.3-7 cycloalkyl, 3-7 membered heterocycloalkyl, 5-6 membered aryl or 5-6 membered heteroaryl, wherein the C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, C.sub.3-7 cycloalkyl, 3-7 membered heterocycloalkyl, 5-6 membered aryl or 5-6 membered heteroaryl is optionally substituted with 1, 2, 3 or 4 R; R is selected from H, halogen, NH.sub.2, CN, OH, C.sub.1-3 alkyl, C.sub.1-3 heteroalkyl, C.sub.3-6 cycloalkyl, 3-6 membered heterocycloalkyl, 5-6 membered aryl or 5-6 membered heteroaryl, wherein the C.sub.1-3 alkyl, C.sub.1-3 heteroalkyl, C.sub.3-6 cycloalkyl, 3-6 membered heterocycloalkyl, 5-6 membered aryl or 5-6 membered heteroaryl is optionally substituted with 1, 2, 3 or 4 R; R is selected from halogen, OH, CN or NH.sub.2; T is selected from N or C(R); and each X is independently selected from N or C; wherein hetero represents 0, 1, 2, 3 or 4 heteroatom(s) or heterogroup(s) independently selected from O, S, N, OH, SH, NH, NH.sub.2, C(O), S(O) or S(O).sub.2.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: R is selected from H, halogen, NH.sub.2, CN, OH, C.sub.1-3 alkyl, C.sub.1-3 alkoxy or C.sub.1-3 alkylamino, wherein the C.sub.1-3 alkyl, C.sub.1-3 alkoxy or C.sub.1-3 alkylamino is optionally substituted with 1, 2, 3 or 4 R.

3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: R is selected from H, F, Cl, Br, I, NH.sub.2, CN, OH, CH.sub.3, CH.sub.2CH.sub.3, N(CH.sub.3).sub.2 or NHCH.sub.3.

4. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: R.sub.1 is selected from H, C.sub.1-3 alkyl, C.sub.1-3 alkyl-OC.sub.1-3 alkyl, C.sub.1-3 alkyl-SC.sub.1-3 alkyl, C.sub.1-3 alkyl-NHC.sub.1-3 alkyl, C.sub.1-6 alkoxy, C.sub.1-3 alkylamino, C.sub.3-6 cycloalkyl or C.sub.3-6 heterocycloalkyl, wherein the C.sub.1-3 alkyl, C.sub.1-3 alkyl-OC.sub.1-3 alkyl, C.sub.1-3 alkyl-SC.sub.1-3 alkyl, C.sub.1-3 alkyl-NHC.sub.1-3 alkyl, C.sub.1-6 alkoxy, C.sub.1-3 alkylamino, C.sub.3-6 cycloalkyl or C.sub.3-6 heterocycloalkyl is optionally substituted with 1, 2, 3 or 4 R; or R.sub.1 is Formula (II): ##STR00063## wherein Formula (II) is optionally substituted with 1, 2, 3 or 4 R.

5. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: R.sub.1 is selected from H, CH.sub.3, CH.sub.2CH.sub.3, CH.sub.2OCH.sub.3 or cyclopropyl, wherein the CH.sub.3, CH.sub.2CH.sub.3, CH.sub.2OCH.sub.3 or cyclopropyl is optionally substituted with 1, 2, 3 or 4 R; or R.sub.1 is Formula (II): ##STR00064## wherein Formula (II) is optionally substituted with 1, 2, 3 or 4 R.

6. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: R.sub.1 is selected from H, CH.sub.3, CH.sub.2CH.sub.3, CHF.sub.2, CF.sub.3, CH.sub.2OCH.sub.3 or cyclopropyl; or R.sub.1 is Formula (II): ##STR00065##

7. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: -L.sub.1-R.sub.1 is selected from H, S(O).sub.2CH.sub.3, S(O).sub.2CH.sub.2CH.sub.3, S(O).sub.2-cyclopropyl, C(O)-cyclopropyl or C(O)CH.sub.2OCH.sub.3, wherein the S(O).sub.2CH.sub.3, S(O).sub.2CH.sub.2CH.sub.3, S(O).sub.2-cyclopropyl, C(O)-cyclopropyl or C(O)CH.sub.2OCH.sub.3 is optionally substituted with 1, 2, 3 or 4 R; or -L.sub.1-R.sub.1 is Formula (II): ##STR00066## wherein Formula (II) is optionally substituted with 1, 2, 3 or 4 R.

8. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: -L.sub.1-R.sub.1 is selected from H, S(O).sub.2CH.sub.3, S(O).sub.2CH.sub.2CH.sub.3, S(O).sub.2CHF.sub.2, S(O).sub.2CF.sub.3, S(O).sub.2-cyclopropyl, C(O)-cyclopropyl or C(O)CH.sub.2OCH.sub.3; or -L.sub.1-R.sub.1 is Formula (II): ##STR00067##

9. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: R.sub.2 is selected from H, NH.sub.2, C.sub.1-3 alkyl, C.sub.1-3 alkyl-OC.sub.1-3 alkyl, C.sub.1-3 alkyl-SC.sub.1-3 alkyl, C.sub.1-3 alkyl-NHC.sub.1-3 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, C.sub.3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein the C.sub.1-3 alkyl, C.sub.1-3 alkyl-OC.sub.1-3 alkyl, C.sub.1-3 alkyl-SC.sub.1-3 alkyl, C.sub.1-3 alkyl-NHC.sub.1-3 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, C.sub.3-6 cycloalkyl or 3-6 membered heterocycloalkyl is optionally substituted with 1, 2, 3 or 4 R.

10. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: R.sub.2 is selected from H, NH.sub.2, CH.sub.3, CH.sub.2OCH.sub.3, cyclopropyl, cyclobutyl or azetidinyl, wherein the CH.sub.3, CH.sub.2OCH.sub.3, cyclopropyl, cyclobutyl or azetidinyl is optionally substituted with 1, 2, 3 or 4 R.

11. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: R.sub.2 is selected from H, NH.sub.2, CH.sub.2CN, CF.sub.3, CH.sub.2OCH.sub.3, cyclopropyl, cyclopropan-3-ol, 3,3-difluorocyclobutyl or azetidin-3-yl.

12. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: -L.sub.2-R.sub.2 is selected from H, NH.sub.2, NHC(O)CH.sub.3, NHC(O)CH.sub.2OCH.sub.3, NHC(O)-cyclopropyl, NHC(O)-cyclobutyl or NHC(O)-azetidin-3-yl, wherein the NHC(O)CH.sub.3, NHC(O)CH.sub.2OCH.sub.3, NHC(O)-cyclopropyl, NHC(O)-cyclobutyl or NHC(O)-azetidin-3-yl is optionally substituted with 1, 2, 3 or 4 R.

13. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: -L.sub.2-R.sub.2 is selected from H, NH.sub.2, NHC(O)CH.sub.2CN, NHC(O)CF.sub.3, NHC(O)CH.sub.2OCH.sub.3, NHC(O)-cyclopropyl, NHC(O)-cyclobutan-3-ol, NHC(O)-3,3-difluorocyclobutyl or NHC(O)-azetidin-3-yl.

14. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: ring A is 1,3,4-triazolyl, imidazolyl, oxazolyl or thiazolyl.

15. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: ##STR00068##

16. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: ##STR00069##

17. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: ##STR00070##

18. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: ##STR00071##

19. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: ##STR00072##

20. The compound according to claim 1, wherein the compound is selected from the group consisting of: ##STR00073## ##STR00074## ##STR00075## ##STR00076## ##STR00077## ##STR00078## ##STR00079##

Description

DETAILED DESCRIPTION OF THE EMBODIMENTS

Preparation of Intermediate Compounds

Preparation of the Intermediates 1-5

(1) ##STR00027##

Step 1: Preparation of tert-butyl 4-(4-amidopyrimidinyl-2-yl) pyrazole-1-carboxylate (2)

(2) 2-chloro-4-aminopyrimidine (3.0 g, 23.2 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) pyrazol-1-carboxylic ester (8.2 g, 27.8 mmol), and potassium carbonate (9.6 g, 69.5 mmol) were dissolved in the mixed solvent of dioxane (30 mL) and water (5 mL). Then Pd(dppf)Cl.sub.2 (1.7 g, 2.3 mmol) was added. It was vacuumed and purged with nitrogen. The reaction mixture was stirred at 80 C. in an oil bath for 2 hours under the protection of nitrogen, and the complete reaction was tracked and determined by TLC. After cooling, the mixture was filtered through diatomaceous earth and filter cake washed with ethyl acetate (100 mL) and tetrahydrofuran (100 mL). The filtrate was dried by anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by using silica gel column chromatography (eluting with petroleum ether/ethyl acetate=2/11/1) to give tert-butyl 4-(4-amidopyrimidinyl-2-yl) pyrazole-1-carboxylate (4.50 g, 59.49% yield) as a pale yellow oil. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =8.50 (s, 1H), 8.17 (s, 1H), 8.07 (d, J=5.8 Hz, 1H), 6.92 (br. s., 2H), 6.30 (d, J=5.8 Hz, 1H), 1.58 (s, 9H). MS (ESI). Calcd. for C.sub.12H.sub.15N.sub.5O.sub.2 [M+H].sup.+ 262, Found 262.

Step 2: Preparation of tert-butyl 4-(4-(3-(ethoxycarbonyl)thiourea)pyrimidin-2-yl)-1H-pyrazole-1-carboxylate (3)

(3) To tert-butyl 4-(4-aminopyrimidinyl-2-yl) pyrazole-1-carboxylate (4.0 g, 15.3 mmol) dissolved in a solution of tetrahydrofuran (40 mL) and dichloromethane (40 mL) was added ethoxycarbonyl isothiocyanate (4.0 g, 30.6 mmol). The reaction mixture was heated to 70 C. and stirred for 16 hours. After TLC showed that the reaction was completed, the mixture was concentrated under reduced pressure, and purified through silica gel column chromatography (eluting with petroleum ether/ethyl acetate=10:12:1) to give tert-butyl 4-(4-(3-(ethoxycarbonyl)thiourea) pyrimidinyl-2-yl)-1H-pyrazole-1-carboxylate (4.00 g, 63.25% yield) as a pale yellow oil. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =12.49 (br. s., 1H), 12.16 (br. s., 1H), 8.78 (d, J=5.8 Hz, 1H), 8.72 (s, 1H), 8.32 (s, 1H), 8.22 (br. s., 1H), 4.27 (q, J=7.1 Hz, 2H), 1.62 (s, 9H), 1.30 (t, J=7.2 Hz, 3H). MS (ESI) Calcd. for C.sub.16H.sub.20N.sub.6O.sub.4S [M+H].sup.+ 393, Found 393.

Step 3: Preparation of 5-(1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-c] pyrimidin-2-amine (Intermediate 1)

(4) To hydroxylamine hydrochloride (3.5 g, 50.9 mmol) dissolved in a solution of methanol (50 mL) and ethanol (50 mL) was added DIEA (4.0 g, 30.6 mmol). After stirring the resulting turbid solution at 26 C. for 1 hour, tert-butyl 4-(4-(3-(ethoxycarbonyl)thiourea) pyrimidin-2-yl)-1H-pyrazole-1-carboxylate was added. Then the reaction mixture was heated to 90 C. for refluxing 3 hours, and TLC showed that the reaction was completed. The reaction mixture was concentrated under reduced pressure, and water (20 mL) was added. The resulting precipitation was filtered, collected, and dried under vacuum to give 5-(1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-c] pyrimidin-2-amine (1.7 g, 82.9% yield) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =13.53 (br. s., 1H), 8.87 (br. s., 1H), 8.47 (br. s., 1H), 8.10 (d, J=6.0 Hz, 1H), 7.23 (d, J=6.0 Hz, 1H), 6.50 (s, 2H). MS (ESI) Calcd. for C.sub.8H.sub.7N.sub.7[M+H].sup.+ 202, Found 202.

Step 4: Preparation of tert-butyl 3-[4-(2-amino-[1,2,4] triazolo[1,5-c] pyrimidin-5-yl) pyrazole-1-yl]-3-(cyanomethyl)azetidine-1-carboxylate (Intermediate 2)

(5) To the Intermediate 1 (500 mg, 2.5 mmol) slightly dissolved in a suspension of acetonitrile (10 mL) was added tert-butyl 3-(cyanomethylene)azetidine-1-carboxylate (600 mg, 3.1 mmol) and DBU (756 mg, 4.97 mmol). The reaction mixture was stirred at 26 C. for 16 hours. After TLC showed that the reaction was completed, the mixture was concentrated under reduced pressure, and purified through silica gel column chromatography (eluting with petroleum ether/ethyl acetate=1:11:3) to give tert-butyl 3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl)pyrazole-1yl]-3-(cyanomethyl) azetidine-1-carboxylate (800 mg, 81.3% yield) as a white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) =9.03 (s, 1H), 8.64 (s, 1H), 8.17 (d, J=6.0 Hz, 1H), 7.24 (d, J=6.0 Hz, 1H), 4.83 (s, 2H), 4.57 (d, J=9.8 Hz, 2H), 4.33 (d, J=9.8 Hz, 2H), 3.36 (s, 2H), 1.49 (s, 9H). MS (ESI) Calcd. for C.sub.18H.sub.21N.sub.9O.sub.2 [M+H].sup.+ 396, Found 396.

Step 5: Preparation of 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl) pyrazole-1-yl]cyclobutylamine-3-yl] acetonitrile (Intermediate 3)

(6) To the Intermediate 2 (500 mL, 1.3 mmol) in DCM (10 mL) was added TFA (4 mL) at 15 C., and at this temperature, the mixture was stirred and reacted for 3 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to give 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl) pyrazole-1-yl]cyclobutylamine-3-yl]acetonitrile (515 mg, 99.9% yield, TFA salt) as a brown solid. MS (ESI) Calcd. for C.sub.13H.sub.13N.sub.9 [M+H].sup.+ 296, Found 296.

Step 6: Preparation of tert-butyl 3-(cyanomethyl)-3-[4-2-(cyclopropylcarbonylamino)-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl]pyrazole-1-yl] azetidine-1-carboxylate (Intermediate 4)

(7) To tert-butyl 3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl)pyrazole-1-yl]-3-(cyanomethyl) azetidine-1-carboxylate (400 mg, 1.0 mmol) slightly dissolved in turbid solution of acetonitrile (8 mL) was added cyclopropanecarboxylic acid chloride (317 mg, 3.0 mmol) and triethylamine (307 mg, 3.0 mmol). The reaction mixture was stirred at 26 C. for 16 hours. After TLC showed that the reaction was completed, a disubstituted product was formed from LC-MS display. The reaction mixture was concentrated under reduced pressure, then methylamine in ethanol solution (27%32%, 3 mL) was added, stirred at 26 C. for 0.5 hour. A monosubstituted target product was formed from LC-MS display. The reaction mixture was concentrated under reduced pressure, and purified through silica gel column chromatography (eluting with petroleum ether/ethyl acetate=1:11:3) to give tert-butyl 3-(cyanomethyl)-3-[4-2-(cyclopropylcarbonylamino)-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl]pyrazole-1-yl]azetidine-1-carboxylate (420 mg, 89.7% yield) as a white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) =9.31 (s, 1H), 9.11 (s, 1H), 8.67 (s, 1H), 8.28 (d, J=6.0 Hz, 1H), 7.42 (d, J=6.0 Hz, 1H), 4.58 (d, J=9.5 Hz, 2H), 4.34 (d, J=9.5 Hz, 2H), 3.37 (s, 2H), 1.50 (s, 9H), 1.31-1.22 (m, 3H), 1.03 (qd, J=3.7, 7.4 Hz, 2H). MS (ESI) Calcd. for C.sub.22H.sub.25N.sub.9O.sub.3 [M+H].sup.+464, Found 464.

Step 7: Preparation of N-[5-[1-[3-(cyanomethyl)azetidine-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c] pyrimidin-2-yl]cyclopropanecarboxamide (Intermediate 5)

(8) To 3-(cyanomethyl)-3-[4-2-(cyclopropylcarbonylamino)-[1,2,4]triazolo[1,5-c] pyrimidin-5-yl]pyrazole-1-yl]azetidine-1-carboxylate (220 mg, 474.7 umol) dissolved in dichloromethane (8 mL) was added trifluoroacetic acid (2 mL). The reaction mixture was stirred at 26 C. for 2 hours. After TLC showed that the reaction was completed, the mixture was concentrated under reduced pressure to give N-[5-[1-[3-(cyanomethyl)azetidine-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]cyclopropanecarboxamide (280 mg, the crude product is directly used in the next step) as a pale yellow solid. MS (ESI) Calcd. for C.sub.17H.sub.17N.sub.9O [M+H].sup.+ 364, Found 364.

Preparation of the Intermediates 6-8

(9) ##STR00028## ##STR00029##

Step 1: Preparation of ethyl-N-[(6-bromo-2-pyridyl)thiocarbamoyl]carbamic acid (2)

(10) To 6-bromopyridine-2-amine (30 mg, 173.4 mmol) dissolved in dichloromethane (400 mL) was added dropwise ethyl isothiocyanate (25.0 g, 190.7 mmol), and the reaction was carried out at 25 C. for 16 hours. After TLC showed that the reaction was completed, the reaction mixture was distilled under reduced pressure. The resulting residue was stirred by using 200 mL of petroleum ether for 30 min, then filtered. The filter cake was collected and dried to give ethyl-N-[(6-bromo-2-pyridyl)thiocarbamoyl]carbamic acid (51 g, 96.7% yield) as a faint red solid. .sup.1H NMR (400 MHz, DMSO-d6) =12.17 (s, 1H), 11.66 (br. s., 1H), 8.65 (d, J=7.54 Hz, 1H), 7.82 (t, J=7.92 Hz, 1H), 7.49 (d, J=7.78 Hz, 1H), 4.22 (q, J=7.18 Hz, 2H), 1.25 (t, J=7.16 Hz, 3H). MS (ESI) Calcd. for C.sub.9H.sub.10BrN.sub.3O.sub.2S [M+H].sup.+ 304, Found 304.

Step 2: Preparation of 5-bromo-[1,2,4] triazolo[1,5-a]pyridin-2-amine (3)

(11) Hydroxylamine hydrochloride (35.2 g, 503.1 mmol), diisopropylethylamine (54.1 g, 419.3 mmol) was dissolved in a mixed solvent of ethanol (500 mL) and methanol (500 mL). After stirring at 25 C. for 1 hour, ethyl-N-[(6-bromo-2-pyridyl)thiocarbamoyl] carbamic acid (51.0 g, 167.7 mmol) was added, and purged with nitrogen for three times. The reaction mixture was heated to 80 C. for the reaction for 3 hours, and then cooled. After the reaction was completed from the TLC monitoring display, the reaction mixture was distilled under reduced pressure. The resulting residue was stirred by using 500 mL of water for 10 min, then filtered. The filter cake was collected and dried to give 5-bromo-[1,2,4]triazolo[1,5-a]pyridin-2-amine (32 g, 85.1% yield) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.30-7.39 (m, 1H), 7.20 (dd, J=6.78, 1.76 Hz, 1H), 6.27 (s, 2H). MS (ESI) Calcd. for C.sub.6H.sub.5BrN.sub.4 [M+H].sup.+ 215, Found 215.

Step 3: Preparation of N-(5-bromo-[1,2,4]triazolo[1,5-a]pyridin-2-yl)cyclopropyl carboxamide (4)

(12) To 5-bromo-[1,2,4] triazolo[1,5-a]pyridin-2-amine (15.00 g, 70.41 mmol) and triethylamine (21.4 g, 211.2 mmol), dissolved in acetonitrile (150 mL) was added dropwise cyclopropanecarboxylic acid chloride (8.8 g, 84.5 mmol) at 0 C. After adding, the mixture was warmed to room temperature for the reaction for 16 hours. After TLC showed that the reaction was completed through monitoring, the reaction mixture was distilled under reduced pressure. The resulting residue was dissolved in an alcoholic solution of methylamine (150 mL), heated to 80 C. for the reaction for 1 hour, cooled, distilled under reduced pressure. The resulting residue was dissolved in mixed solution of water (100 mL) and ethyl acetate (200 mL), and the layers were separated and extracted. The combined organic phase was dried with anhydrous sodium sulfate and filtered, and the filtrate was distilled under reduced pressure. The resulting crude product was purified through silica gel column chromatography (eluting with ethyl acetate/petroleum ether=070%) to give N-(5-bromo-[1,2,4]triazolo[1,5-a]pyridin-2-yl)cyclopropyl carboxamide (7.2 g, 56.64% yield) as a pale yellow solid. .sup.1H NMR (400 MHz, DMSO-d6) =11.20 (br. s., 1H), 7.68-7.73 (m, 1H), 7.52-7.58 (m, 1H), 7.46-7.51 (m, 1H), 1.96-2.09 (m, 1H), 0.82 (d, J=6.28 Hz, 4H). MS (ESI) Calcd. for C.sub.10H.sub.9BrN.sub.4O [M+H].sup.+ 282, Found 282.

Step 4: Preparation of N-[5-(1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl]cyclopropane carboxamide (Intermediate 6)

(13) To N-(5-bromo-[1,2,4]triazolo[1,5-a] pyridin-2-yl)cyclopropane carboxamide (3.0 mg, 10.67 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (2.4 g, 12.9 mmol) and potassium carbonate (3.7 g, 26.7 mmol), dissolved in a mixed solution of dioxane (30 mL) and water (5 mL), was added Pd(dppf)Cl.sub.2 (260 mg) in the nitrogen atmosphere. The resulting mixture was heated to 110 C. for the reaction for 3 hours, then cooled to room temperature. After TLC showed that raw material was completely reacted, the reaction mixture was filtered. The filtrate was washed with water (150 mL) and extracted with ethyl acetate (150 mL3). The combined organic phase was dried with anhydrous sodium sulfate and filtered, and the filtrate was distilled under reduced pressure. The resulting crude product was purified through silica gel column chromatography (eluting with ethyl acetate/petroleum ether=50100%) to give N-[5-(1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl] cyclopropane carboxamide (2.1 g, 62.4% yield) as a gray solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =13.37 (br. s., 1H), 11.15 (br. s., 1H), 8.96 (s, 1H), 8.53 (s, 1H), 7.57-7.72 (m, 2H), 7.51 (d, J=8.28 Hz, 1H), 2.06 (br. s., 1H), 0.78-0.91 (m, 4H). MS (ESI) Calcd. for C.sub.13H.sub.12N.sub.6O [M+H].sup.+ 269, Found 269.

Step 5: Preparation of tert-butyl 3-(cyanomethyl)-3-[4-[2-(cyclopropylcarbonylamino)-[1,2,4]triazolo[1,5-a]pyridin-5-yl]pyrazole-1-yl]azetidine-1-carboxylate (Intermediate 7)

(14) To N-[5-(1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl] cyclopropane carboxamide (200 mg, 745.5 umol), tert-butyl 3-(cyanomethylene)azetidine-1-carboxylate (144.8 mg, 745.5 umol), dissolved in acetonitrile (5 mL), was added DBU (340.49 mg, 2.3 mmol) and reacted at room temperature for 16 hours. After LCMS showed that raw material was completely reacted, the reaction mixture was distilled under reduced pressure. The resulting residue was dissolved in mixed solution of water (20 mL) and ethyl acetate (20 mL), and the layers were separated and extracted. The combined organic phase was dried with anhydrous sodium sulfate and filtered, and the filtrate was distilled under reduced pressure. The resulting crude product was purified through preparative TLC (pure ethyl acetate) to give tert-butyl 3-(cyanomethyl)-3-[4-[2-(cyclopropylcarbonylamino)-[1,2,4]triazolo[1,5-a]pyridin-5-yl]pyrazole-1-yl]azetidine-1-carboxylate (170 mg, 44.4% yield) as a pale yellow solid. MS (ESI) Calcd. for C.sub.23H.sub.26N.sub.8O.sub.3 [M+H].sup.+ 463, Found 463.

Step 6: Preparation of N-[5-[1-[3-(cyanomethyl) azetidine-3-yl] pyrazole-4-yl]-[1,2,4]triazolo[1,5-a]pyridin-2-yl] cyclopropane carboxamide (Intermediate 8)(WX00)

(15) To tert-butyl 3-(cyanomethyl)-3-[4-2-(cyclopropylcarbonylamino)-[1,2,4]triazolo[1,5-a]pyridin-5-yl] pyrazole-1-yl]azetidine-1-carboxylate (150 mg, 324.3 umol) dissolved in 5 mL of dichloromethane (5 mL) was added trifluoroacetic acid (1 mL) at room temperature for the reaction for 2 hours. After LCMS showed that the reaction was completely reacted, the reaction mixture was distilled under reduced pressure to give crude product of N-[5-[1-[3-(cyanomethyl) azetidine-3-yl] pyrazole-4-yl]-[1,2,4] triazolo[1,5-a]pyridin-2-yl]cyclopropane carboxamide (100 mg) as a yellow oil which was directly used in the next step. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =11.16 (br. s., 1H), 9.18 (s, 1H), 8.71 (s, 1H), 7.75-7.63 (m, 2H), 7.61-7.50 (m, 1H), 4.00 (d, J=9.0 Hz, 2H), 3.71 (d, J=9.0 Hz, 2H), 3.57 (s, 2H), 2.13 (br. s., 1H), 0.95-0.81 (m, 4H). MS (ESI) Calcd. for C.sub.18H.sub.18N.sub.8O [M+H].sup.+ 363, Found 363.

Preparation of the Intermediates 9

(16) ##STR00030##

Step 1: Preparation of tert-butyl 3-(cyanomethyl)azetine-1-carbonate (1)

(17) Under cooling in ice bath, to sodium-hydrogen (1.2 g, 30.7 mmol) in tetrahydrofuran (50 mL) was added dropwise cyanomethyl diethyl phosphite (5.7 g, 32.1 mmol) in tetrahydrofuran (50 mL). After adding dropwise, the mixture was stirred at 25 C. for 1 hour, then cooled to 0 C. Then tert-butyl 3-azetidinone-1-carbonate (5.0 g, 29.2 mmol) in tetrahydrofuran (50 mL) was added dropwise within 1 hour. The mixture is stirred at 25 C. for the reaction for 16 hours. After the reaction was completed, the reaction mixture was quenched with water (80 mL) and extracted with ethyl acetate (80 mL3). The combined organic phase was washed with saturated salt water and dried with anhydrous sodium sulfate to give crude product of tert-butyl 3-(cyanomethyl) azetine-1-carbonate (5.2 g, 78.0% yield) as a yellow solid, the crude product was directly used in the next step without further purification. .sup.1H NMR (400 MHz, CDCl.sub.3) =5.38 (t, J=2.5 Hz, 1H), 4.73-4.68 (m, 2H), 4.61 (td, J=2.4, 4.3 Hz, 2H), 1.45 (s, 9H). MS (ESI) Calcd. for C.sub.10H.sub.14N.sub.2O.sub.2 [M+H].sup.+ 195, Found 195.

Step 2: Preparation of 2-(azetine-3-yl)methyl cyanide(2)

(18) Tert-butyl 3-(cyanomethyl) azetine-1-carbonate (5.2 g, 26.8 mmol) was soaked with a small amount of ethyl acetate (5 mL). After stirring well, hydrochloride ethyl acetate (150 mL) was added at 0 C. and stirred at 0 C. for 1 hour. After TLC showed that the reaction was completed (petroleum ether/ethyl acetate=5:1), the resulting yellow suspension was filtered. The resulting solid was washed with a small amount of cold ethyl acetate (5 mL2) and dried under vacuum to give 2-(azetine-3-yl) methyl cyanide (2.8 g, 80.0% yield) as a white solid. .sup.1H NMR (400 MHz, D.sub.2O) =5.69-5.65 (m, 1H), 4.95 (d, J=2.5 Hz, 2H), 4.88 (br. s., 2H). MS (ESI) Calcd. for C.sub.5H.sub.6N.sub.2 [M+H].sup.+ 95, Found 95.

Step 3: Preparation of 2-(1-(ethyl sulfonyl)azetine-3-yl)methyl cyanide (Intermediate 9)

(19) To a solution of 2-(azetine-3-yl) methyl cyanide (2.8 g, 21.4 mmol) and DIPEA (8.3 g, 64.3 mmol) in dichloromethane (30 mL) was added dropwise ethanesulfonyl chloride (4.1 g, 32.1 mmol) at 0 C. under the protection of nitrogen, and the temperature was kept below 2 C. when dropping. The reaction mixture was stirred at 25 C. for reacting for 16 hours. TLC showed that the reaction was completed (petroleum ether/ethyl acetate=1:1). After the reaction mixture was quenched with water, extracted with dichloromethane (30 mL2). The combined organic phase was washed with saturated salt water (20 mL2), dried with anhydrous sodium sulfate, filtered, and spun dry. The residue was purified through column chromatography (dichloromethane/ethyl acetate=3/1) to give 2-(1-(ethylsulfonyl)azetine-3-yl)methyl cyanide (1.4 g, 33.0% yield) as a pale yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) =5.50-5.41 (m, 1H), 4.79 (d, J=3.0 Hz, 2H), 4.71 (d, J=2.5 Hz, 2H), 3.06 (q, J=7.4 Hz, 2H), 1.40 (t, J=7.4 Hz, 3H). MS (ESI) Calcd. for C.sub.7H.sub.10N.sub.2O.sub.2S [M+H].sup.+187, Found 187.

Step 3: Preparation of 2-(1-cyclopropylsulfonylazetidine-3-alkenyl)acetonitrile (Intermediate 10)

(20) Intermediate 10 was prepared in the same method as Intermediate 9. 2-(1-cyclopropylsulfonylazetidine-3-alkenyl)acetonitrile (1.5 g) was a pale yellow solid, MS (ESI) Calcd. for C.sub.7H.sub.10N.sub.2O.sub.2S [M+H].sup.+ 199, Found 199.

Example 1

(21) ##STR00031##

Step 1: Preparation of 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl) pyrazole-1-yl]-1-ethylsulfonylazetidine-3-yl] acetonitrile (1)

(22) To a suspension of Intermediate 1 (150 mg, 745.6 umol) slightly dissolved in acetonitrile (4 mL) and DMF (2 mL) was added Intermediate 9 (208 mg, 1.1 mmol) and DBU (227 mg, 1.5 mmol). The reaction mixture was stirred at 26 C. for 16 hours. LC-MS showed that the reaction was completed. The precipitated solid was filtered, collected, washed with cold acetonitrile (5 mL), and dried under reduced pressure to give 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl) pyrazole-yl]-1-ethylsulfonylazetidine-3-yl] acetonitrile (200 mg, 69.2% yield) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =9.16 (s, 1H), 8.69 (s, 1H), 8.16 (d, J=6.3 Hz, 1H), 7.31 (d, J=6.0 Hz, 1H), 6.56 (s, 2H), 4.53 (d, J=9.0 Hz, 2H), 4.28 (d, J=9.0 Hz, 2H), 3.70 (s, 2H), 3.25 (q, J=7.4 Hz, 2H), 1.25 (t, J=7.4 Hz, 3H). MS (ESI) Calcd. for C.sub.15H.sub.17N.sub.9O.sub.2S [M+H].sup.+ 388, Found 388.

Step 2: Preparation of N-[5-[1-[3-(cyanomethyl)-1-ethylsulfonylazetidine-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]cyclopropanecarboxamide(WX01)

(23) To a suspension of 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl) pyrazole-yl]-1-ethylsulfonyl azetidine-3-yl] acetonitrile (100 mg, 258.1 umol) slightly dissolved in acetonitrile (2 mL) and tetrahydrofuran (1 mL) was added cyclopropanecarboxylic acid chloride (80.9 mg, 774.4 umol) and triethylamine (78 mg, 774.4 mmol). The reaction mixture was stirred at 26 C. for 16 hours. TLC showed that the reaction was completed, and LC-MS showed that all was produced as a di-substituted product. After the reaction mixture was concentrated under reduced pressure, methylamine in ethanol solution (27%-32%, 3 mL) was added and stirred at 26 C. for reacting for 0.5 hour. LC-MS showed that all was produced as a mono-substituted product. The reaction mixture was concentrated under reduced pressure and purified through preparative HPLC (alkaline condition) to give N-[5-[1-[3-(cyanomethyl)-1-ethylsulfonylazetidine-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]cyclopropanecarboxamide (25 mg, 21.1% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) =11.43 (br. s., 1H), 9.25 (s, 1H), 8.82 (s, 1H), 8.32 (d, J=6.0 Hz, 1H), 7.60 (d, J=6.3 Hz, 1H), 4.50 (d, J=9.0 Hz, 2H), 4.28 (d, J=9.0 Hz, 2H), 3.70 (s, 2H), 3.24 (q, J=7.3 Hz, 2H), 2.18-2.02 (m, 1H), 1.23 (t, J=7.3 Hz, 3H), 0.95-0.80 (m, 4H). MS (ESI) Calcd. for C.sub.19H.sub.21N.sub.9O.sub.3S [M+H].sup.+ 456, Found 456.

Example 2

(24) ##STR00032##

Step 1: Preparation of N-[5-[1-[3-(cyanomethyl)-1-(trifluoromethylsulfonyl) azetidine-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]cyclopropanecarboxamide(WX02)

(25) To a suspension of Intermediate 5 (100 mg, 209.5 umol) slightly dissolved in dichloromethane (3 mL) was added trifluoromethanesulfonyl chloride (53 mg, 314.2 umol) and triethylamine (106 mg, 1.1 mmol). The reaction mixture was stirred at 26 C. for 16 hours. LC-MS showed that the reaction was completed. The reaction mixture was concentrated under reduced pressure and purified through preparative HPLC (alkaline condition) to give N-[5-[1-[3-(cyanomethyl)-1-(trifluoromethylsulfonyl) azetidine-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]cyclopropanecarboxamide (25 mg, 24.1% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) =11.45 (br. s., 1H), 9.27 (s, 1H), 8.90 (s, 1H), 8.35 (d, J=6.0 Hz, 1H), 7.63 (d, J=6.3 Hz, 1H), 4.90 (d, J=9.0 Hz, 2H), 4.72 (d, J=9.0 Hz, 2H), 3.85 (s, 2H), 1.30-1.23 (m, 1H), 0.97-0.87 (m, 4H). MS (ESI) Calcd. for C.sub.18H.sub.16F.sub.3N.sub.9O.sub.3S [M+H].sup.+ 496, Found 496.

Example 3

(26) ##STR00033##

Step 1: Preparation of 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl) pyrazole-1-yl]-1-cyclopropylsulfonyl-azetidine-3-yl] acetonitrile (2)

(27) To a suspension of Intermediate 1 (150 mg, 745.6 umol) dissolved in acetonitrile (4 mL) was added Intermediate 10 (192 mg, 969.2 umol) and DBU (227 mg, 1.5 mmol). The reaction mixture was stirred at 26 C. for 16 hours. TLC showed that the reaction was completed. The precipitated solid was filtered, collected, washed with cold acetonitrile (5 mL), and dried under reduced pressure to give 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl)pyrazole-1-yl]-1-cyclopropylsulfonyl-azetidine-3-yl] acetonitrile (200 mg, 67.2% yield) as a white solid. H NMR (400 MHz, DMSO-d.sub.6) =9.18 (s, 1H), 8.70 (s, 1H), 8.15 (d, J=6.3 Hz, 1H), 7.30 (d, J=6.0 Hz, 1H), 6.56 (s, 2H), 4.59 (d, J=9.3 Hz, 2H), 4.33 (d, J=9.3 Hz, 2H), 3.70 (s, 2H), 2.90-2.82 (m, 1H), 1.09-1.03 (m, 2H), 1.03-0.96 (m, 2H). MS (ESI) Calcd. for C.sub.16H.sub.17N.sub.9O.sub.2S [M+H].sup.+ 400, Found 400.

Step 2: Preparation of N-[5-[1-[3-(cyanomethyl)-1-cyclopropylsulfonyl-azetidine-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]cyclopropanecarboxamide (WX3)

(28) To a suspension of 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl)pyrazole-1-yl]-1-cyclopropylsulfonyl-azetidine-3-yl]acetonitrile (80 mg, 200.3 umol) slightly dissolved in acetonitrile (2 mL) was added cyclopropanecarboxylic acid chloride (63 mg, 600.9 umol) and triethylamine (61 mg, 600.9 umol). The reaction mixture was stirred at 26 C. for 16 hours and at 80 C. for 3 hours. LC-MS showed that all was produced as a mixture of mono- and di-substituted product. After the reaction mixture was concentrated under reduced pressure, methylamine in ethanol solution (27%-32%, 3 mL) was added and stirred at 26 C. for 0.5 hour. LC-MS showed that all was produced as a mono-substituted product. The reaction mixture was concentrated under reduced pressure and purified through preparative HPLC (alkaline condition) to give N-[5-[1-[3-(cyanomethyl)-1-cyclopropylsulfonyl-azetidine-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]cyclopropanecarboxamide (60 mg, 64.1% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) =11.43 (s, 1H), 9.27 (s, 1H), 8.81 (s, 1H), 8.32 (d, J=6.0 Hz, 1H), 7.60 (d, J=6.0 Hz, 1H), 4.57 (d, J=9.3 Hz, 2H), 4.32 (d, J=9.3 Hz, 2H), 3.70 (s, 2H), 2.92-2.79 (m, 1H), 2.07 (d, J=13.6 Hz, 1H), 1.07-0.96 (m, 4H), 0.91-0.83 (m, 4H). MS (ESI) Calcd. for C.sub.20H.sub.21N.sub.9O.sub.3S [M+H].sup.+ 468, Found 468.

Example 4

(29) ##STR00034##

Step 1: Preparation of 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl) pyrazole-1-yl]-1-methylsulfonyl-cyclobutylamine-3-yl] acetonitrile (WX04)

(30) To Intermediate 3 (1.0 g, 2.4 mmol, TFA salt) and triethylamine (617 mg, 6.1 mmol), dissolved in solution of DCM (50 mL) was added dripwise MsCl (307 mg, 2.7 mmol) at 15 C. After dropping, the reaction mixture was stirred at 15 C. for 2 hours. After the reaction was completed, the reaction mixture was concentrated to dry. The resulting solid was separated and purified through preparative HPLC (alkaline condition) to give 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl)pyrazole-1-yl]-1-methylsulfonyl-cyclobutylamine-3-yl]acetonitrile (800 mg, 87.8% yield). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) =9.17 (s, 1H), 8.70 (s, 1H), 8.15 (d, J=6.0 Hz, 1H), 7.30 (d, J=6.0 Hz, 1H), 6.56 (brs, 2H), 4.55 (d, J=9.2 Hz, 2H), 4.31 (d, J=9.2 Hz, 2H), 3.69 (s, 2H), 3.14 (s, 3H). MS (ESI) Calcd. for C.sub.14H.sub.12F.sub.3N.sub.9O.sub.2S [M+H].sup.+ 428, Found 428.

Example 5

(31) ##STR00035##

Step 1: Preparation of 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl) pyrazole-1-yl]-1-(trifluoromethylsulfonyl)cyclobutylamine-3-yl]acetonitrile (WX05)

(32) To a solution of Intermediate 3 (515 mg, 1.7 mmol) and TEA (264 mg, 2.6 mmol) dissolved in DCM (10 mL), was added dropwise trifluoromethanesulfonyl chloride (323 mg, 1.9 mmol) at 15 C. under the protection of nitrogen. After dropping, the reaction mixture was stirred at 15 C. for 3 hours. After the reaction was completed, the mixture was concentrated to dry. The resulting solid was beat with water, and filtered, the filter cake was dried to give 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl)pyrazole-1-yl]-1-(trifluoro methylsulfonyl)cyclobutylamine-3-yl]acetonitrile (700 mg, 94.1% yield) as a white solid. .sup.1H-NMR (400 MHz, DMSO-d.sub.6) =9.18 (s, 1H), 8.74 (s, 1H), 8.15 (d, J=6.0 Hz, 1H), 7.30 (d, J=6.0 Hz, 1H), 6.55 (brs, 2H), 4.91 (d, J=9.2 Hz, 2H), 4.70 (d, J=9.2 Hz, 2H), 3.82 (s, 2H). MS (ESI) Calcd. for C.sub.14H.sub.12F.sub.3N.sub.9O.sub.2S [M+H].sup.+ 428, Found 428.

Step 2: Preparation of N-[5-[1-[3-(cyanomethyl)-1-(trifluoromethylsulfonyl) cyclobutylamine-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]cyclobutylamine-3-formamide(WX06)

(33) To DCM (10 mL) solution of N-tert-butyl-formylcyclobutylamine-3-carboxylic acid (198 mg, 982.8 umol) and DMF (100 uL) was added dropwise oxalyl chloride (156 mg, 1.23 mmol) in DCM (2 mL) at 0 C. under the protection of nitrogen. After dropping, the reaction mixture was stirred at 0 C. for 2 hours. Then the reaction mixture was concentrated under reduced pressure to dry. The resulting liquid was dissolved in DCM (2 mL), and was added dropwise through a injector to 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl)pyrazole-1-yl]-1-(trifluoromethylsulfonyl) cyclobutylamine-3-yl]acetonitrile (350 mg, 818.9 umol) in DCM (10 mL) at 0 C. under the protection of nitrogen. After dropping, the reaction mixture was stirred at 0 C. for 2 hours. The reaction mixture was directly purified through preparative thin-layer chromatography (DCM/MeOH=10/1) to give Boc protected product (10 mg). The product was dissolved in DCM (2 mL), to which the TFA (2 mL) was added at 15 C. The resulting mixture was stirred at 15 C. for 1 hour. After the reaction was completed, the mixture was concentrated and dried. The resulting solid was separated and purified through preparative HPLC (alkaline method) to give N-[5-[1-[3-(cyanomethyl)-1-(trifluoromethylsulfonyl)cyclobutylamine-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl] cyclobutylamine-3-formamide (2 mg, 0.42% yield). MS (ESI) Calcd. for C.sub.18H.sub.17F.sub.3N.sub.10O.sub.3S [M+H].sup.+ 511, Found 511.

Example 6

(34) ##STR00036##

Step 1: Preparation of tert-butyl-3-(cyanomethyl)-3-[4-[2-[(3-hydroxycyclobutane formoxyl)amino]-[1,2,4] triazolo[1,5-c]pyrimidin-5-yl] pyrazole-1-yl]cyclobutane-1-formate (1)

(35) To pyridine (10 mL) was added the mixture of Intermediate (300 mg, 758.7 umol), 3-hydroxycyclobutane formic acid (106 mg, 910.4 umol) and EDCI (218 mg, 1.1 mmol), and the resulting mixture was heated to reflux for 16 hours under the protection of nitrogen. After concentrating to dry, the remaining solid was purified through preparative thin-layer chromatography (DCM/MeOH=10/1) to give tert-butyl-3-(cyanomethyl)-3-[4-[2-[(3-hydroxycyclobutaneformoxyl)amino]-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl] pyrazole-1-yl]cyclobutane-1-formate (36 mg, 9.61% yield) as a white solid. MS (ESI) Calcd. for C.sub.23H.sub.27N.sub.9O.sub.4 [M+H].sup.+ 494, Found 494.

Step 2: Preparation of N-[5-[1-[3-(cyanomethyl) cyclobutylamine-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]-3-hydroxy-cyclobutane formamide (2)

(36) To a solution of tert-butyl-3-(cyanomethyl)-3-[4-[2-[(3-hydroxycyclobutaneformoxyl) amino]-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl]pyrazole-1-yl]cyclobutane-1-formate (36 mg, 72.9 umol) in dichloromethane (2.00 mL) was added dropwise TFA (1 mL) at 15 C. The resulting mixture was stirred at 15 C. for 30 min. LCMS showed that the reaction was completed. The reaction mixture was concentrated at 30 C. until dry to give N-[5-[1-[3-(cyanomethyl)cyclobutylamine-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]-3-hydroxy-cyclobutane formamide (37 mg, 99.9% yield, TFA salt) as a yellow solid. MS (ESI) Calcd. for C.sub.18H.sub.19N.sub.9O.sub.2 [M+H].sup.+ 394, Found 394.

Step 3: Preparation of N-[5-[1-[3-(cyanomethyl)-1-(trifluoromethylsulfonyl) cyclobutylamine-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]-3-hydroxy-cyclobutane formamide(WX07)

(37) To a solution of N-[5-[1-[3-(cyanomethyl)cyclobutylamine-3-yl] pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]-3-hydroxy-cyclobutane formamide (15 mg, 29.6 umol) and triethylamine (9 mg, 88.7 umol) in DCM (5.00 mL) was added dropwise a solution of trifluoromethanesulfonyl chloride (7 mg, 44.34 umol) in DCM (1 mL) at 20 C. under the protection of nitrogen. After adding, the mixture was stirred at 20 C. for 1 hour. After the reaction was completed, the reaction mixture was concentrated to dry. The resulting solid was separated and purified through preparative HPLC (0.1% of NH.sub.4OH was used as an additive) to give N-[5-[1-[3-(cyanomethyl)-1-(trifluoromethylsulfonyl)cyclobutylamine-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]-3-hydroxy-cyclobutane formamide (8 mg, 51.50% yield). .sup.1H-NMR (400 MHz, MeOD-d.sub.4) =9.29 (s, 1H), 8.73 (s, 1H), 8.32 (d, J=6.4 Hz, 1H), 7.49 (d, J=6.0 Hz, 1H), 5.00 (d, J=9.6 Hz, 2H), 4.70 (d, J=9.2 Hz, 2H), 4.05-4.13 (m, 1H), 3.70 (s, 2H), 2.90 (brs, 1H), 2.50-2.66 (m, 2H), 2.20-2.35 (m, 2H). MS (ESI) Calcd. for C.sub.19H.sub.8F.sub.3N.sub.9O.sub.4S [M+H].sup.+ 526, Found 526.

Example 7

(38) ##STR00037##

Step 1: Preparation of N-(5-(1-(3-(cyanomethyl)-1-(trifluoromethyl sulfonyl)azetidine-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl) cyclopropane formamide

(39) To Intermediate 8 (100 mg, 275.9 umol) and triethylamine (84 mg, 827.9 umol), dissolved in dichloromethane (5 mL) was dropwise added slowly trifluoromethanesulfonyl chloride (56 mg, 331.4 umol) at 0 C. After adding, the mixture was warmed to room temperature for reacting for 16 hours. After LCMS showed that the reaction was completed, the reaction mixture was diluted with water (20 mL), extracted with dichloromethane (20 mL3). The combined organic phase was dried with anhydrous sodium sulfate and filtered, and the filtrate was distilled under reduced pressure. The resulting residue was purified through thin layer chromatography (ethyl acetate) to give N-(5-(1-(3-(cyanomethyl)-1-(trifluoromethylsulfonyl)azetidine-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)cyclopropane formamide (WX09, 45 mg, 31.33% yield), .sup.1H NMR (400 MHz, METHANOL-d.sub.4) =9.21 (s, 1H), 8.59 (s, 1H), 7.71-7.77 (m, 1H), 7.60 (dd, J=14.44, 8.16 Hz, 2H), 5.00 (d, J=9.04 Hz, 2H), 4.70 (d, J=9.04 Hz, 2H), 3.68 (s, 2H), 1.28-1.39 (m, 1H), 1.11 (quin, J=3.84 Hz, 2H) 0.97-1.04 (m, 2H). MS (ESI) Calcd. for C.sub.19H.sub.17N.sub.8O.sub.3F.sub.3S [M+H].sup.+ 495, Found 495.

(40) Preparation of WX08: N-(5-(1-(3-(cyanomethyl)-1-(cyclopropylsulfonyl)azetidine-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)cyclopropane formamide (WX08) was prepared using the preparation method similar to WX09 (Step 1). .sup.1H NMR (400 MHz, METHANOL-d.sub.4) =9.23 (s, 1H), 8.58 (s, 1H), 7.71-7.76 (m, 1H), 7.58-7.65 (m, 2H), 4.70 (d, J=9.28 Hz, 2H), 4.39 (d, J=9.04 Hz, 2H), 3.64 (s, 2H), 2.72 (dt, J=12.74, 6.31 Hz, 1H), 1.78 (d, J=7.04 Hz, 1H), 1.08-1.14 (m, 6H), 1.00 (dd, J=7.28, 3.26 Hz, 2H). MS (ESI) Calcd. for C.sub.21H.sub.22N.sub.8O.sub.3S [M+H].sup.+ 467, Found 467.

Example 8

(41) ##STR00038##

Step 1: Preparation of N-[5-[1-[3-(cyanomethyl)-1-(2-methoxyacetyl)azetidine-3-]pyrazole-4-]-[1,2,4]triazoloe[1,5-c]pyrimidine2-] cyclopropane carboxamide

(42) 2-Methoxyacetic acid (11 mg, 128.4 umol) was dissolved in mixture solvent of DCM/DMF (6 mL, 5:1). HOBt (35 mg, 256.9 umol) and EDCI (49 mg, 256.9 umol) was added to the mixture in order. The resulting mixture was stirred for reacting for 1 hour. Intermediate 5 (70 mg, 192.6 umol) and DIEA (50 mg, 385.3 umol) were added and stirred at 15 C. for reacting for 12 hours. LC-MS showed that the raw material was completely reacted to the target product. The reaction mixture was concentrated under reduced pressure to remove DCM and DMF and separated through preparative HPLC (alkalinity) to give N-[5-[1-[3-(cyanomethyl)-1-(2-methoxyacetyl)azetidine-3-]pyrazole-4-]-[1,2,4]triazoloe[1,5-c]pyrimidine2-] cyclopropane carboxamide (30 mg, 53.7% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) =9.26 (s, 1H), 8.80 (s, 1H), 8.34 (d, J=6.0 Hz, 1H), 7.61 (d, J=6.3 Hz, 1H), 4.81 (d, J=10.0 Hz, 1H), 4.61 (d, J=10.0 Hz, 1H), 4.46 (d, J=10.5 Hz, 1H), 4.33 (d, J=10.3 Hz, 1H), 4.01 (s, 2H), 3.72 (s, 2H), 3.32 (s, 3H), 2.12 (br. s., 1H), 0.98-0.84 (m, 4H). MS (ESI) Calcd. for C.sub.20H.sub.21N.sub.9O.sub.3 [M+H].sup.+ 436, Found 436.

Example 9

(43) ##STR00039##

Step 1: Preparation of N-[5-[1-[3-(cyanomethyl)-1-(cyclopropylcarbonyl)azetidine-3-]pyrazole-4-]-[1,2,4]triazoloe[1,5-c]pyrimidine2-] cyclopropane carboxamide

(44) Intermediate 5 (79 mg, 216.3 umol) was dissolved in dichloromethane (3 mL), and DIEA (84 mg, 648.9 umol) was added. Then cyclopropionyl chloride (27 mg, 259.6 umol) was added though a syringe within 5 min, and the reaction mixture was stirred at 15 C. for reacting for 3 hours. LC-MS showed that the raw material was completely reacted to the target product. The reaction mixture was concentrated under reduced pressure to remove DCM and DMF and separated through preparative HPLC (alkalinity) to give N-[5-[1-[3-(cyanomethyl)-1-(cyclopropylcarbonyl) azetidine-3-]pyrazole-4-]-[1,2,4]triazoloe[1,5-c]pyrimidine2-] cyclopropane carboxamide (50 mg, 53.6% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) =9.25 (s, 1H), 8.83-8.78 (m, 1H), 8.32 (dd, J=3.8, 6.0 Hz, 1H), 7.60-7.52 (m, 1H), 4.88 (d, J=9.3 Hz, 1H), 4.68 (d, J=9.5 Hz, 1H), 4.44 (d, J=10.5 Hz, 1H), 4.29 (d, J=10.3 Hz, 1H), 3.72 (d, J=5.5 Hz, 2H), 3.13 (br. s., 1H), 1.69-1.55 (m, 1H), 0.98-0.85 (m, 4H), 0.77 (br. s., 4H).sub. MS (ESI) Calcd. for C.sub.21H.sub.21N.sub.9O.sub.2 [M+H].sup.+ 432, Found 432.

Example 10

(45) ##STR00040##

Step 1: Preparation of N-[5-[1-[3-(cyanomethyl)-1-methanesulfonyl-azetidine-3-]pyrazole-4-]-[1,2,4]triazoloe[1,5-c]pyrimidine2-] cyclopropane carboxamide(WX12)

(46) Intermediate 5 (100 mg, 275.2 umol) was suspended in dichloromethane (8 mL). DIEA (107 mg, 825.6 umol) and MsCl (140 mg, 1.2 mmol) was added in order. The reaction mixture was stirred at 15 C. for reacting for 2 hours. LC-MS showed that the raw material was completely reacted, and the target product was detected. The reaction mixture was concentrated under reduced pressure to remove DCM and separated through preparative HPLC (alkalinity) to give N-[5-[1-[3-(cyanomethyl)-1-methanesulfonyl-azetidine-3-]pyrazole-4-]-[1,2,4]triazoloe[1,5-c]pyrimidine2-]cyclopropane carboxamide (39 mg, 31.1% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) =9.28 (s, 1H), 8.86 (s, 1H), 8.35 (d, J=6.3 Hz, 1H), 7.63 (d, J=6.3 Hz, 1H), 6.08 (br. s., 1H), 4.55 (d, J=9.5 Hz, 2H), 4.33 (d, J=9.3 Hz, 2H), 3.72 (s, 2H), 3.16 (s, 3H), 2.10 (d, J=14.8 Hz, 1H), 1.01-0.79 (m, 4H). MS (ESI) Calcd. for C.sub.18H.sub.19N.sub.9O.sub.3S [M+H].sup.+ 442, Found 442.

Example 11

(47) ##STR00041##

Step 1: Preparation of N-[5-[1-[3-(cyanomethyl)-1-(difluoromethylsulfonyl)-azetidine-3-]pyrazole-4-]-[1,2,4]triazoloe[1,5-c]pyrimidine2-]cyclopropane carboxamide (WX13)

(48) Intermediate 5 (100 mg, 275.2 umol) was suspended in dichloromethane (8 mL). DIEA (178 mg, 1.4 mmol) and difluoromethylsulfonyl chloride (62 mg, 412.8 mmol) was added in order. The reaction mixture was stirred at 15 C. for reacting for 12 hours. LC-MS showed that the raw material was completely reacted, and the target product was detected. The reaction mixture was concentrated under reduced pressure to remove DCM. The residue was diluted with DMF and MeOH to a solution (5 mL), and separated through preparative HPLC (alkalinity) to give N-[5-[1-[3-(cyanomethyl)-1-(difluoromethylsulfonyl)-azetidine-3-]pyrazole-4-]-[1,2,4]triazolo[1,5-c]pyrimidin2-]cyclo propyl carboxamide (8 mg, 6.1% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) =11.46 (br. s., 1H), 9.27 (s, 1H), 8.87 (s, 1H), 8.34 (d, J=6.3 Hz, 1H), 7.63 (d, J=6.0 Hz, 1H), 7.42-7.06 (m, 1H), 4.79 (d, J=9.0 Hz, 2H), 4.67-4.49 (m, 2H), 3.78 (s, 2H), 2.26-1.96 (m, 1H), 0.97-0.84 (m, 4H). MS (ESI) Calcd. for C.sub.18H.sub.17F.sub.2N.sub.9O.sub.3S [M+H].sup.+ 478, Found 478.

Example 12

(49) ##STR00042##

Step 1: Preparation of N-[5-[1-[3-(cyanomethyl)-1-(difluoromethylsulfonyl) azetidine-3-]pyrazole-4-]-[1,2,4]triazolo[1,5-c]pyridin-2-]cyclopropyl carboxamide (WX14)

(50) Intermediate 8 (300 mg, 629.7 umol, trifluoroacetate) was suspended in DCM (4 mL). DMAP (8 mg, 63 umol), DIEA (407 mg, 3.2 mmol) and difluoromethylsulfonyl chloride (142 mg, 944.6 umol) was added in order. The reaction mixture was stirred at 15 C. for reacting for 12 hours. LC-MS showed that the raw material was completely reacted, and the target product was detected. The reaction mixture was concentrated under reduced pressure to remove DCM. The residue was diluted with DMF and MeOH to a solution (5 mL), and separated through preparative HPLC (alkalinity) to give N-[5-[1-[3-(cyanomethyl)-1-(difluoromethylsulfonyl)azetidine-3-]pyrazole-4-]-[1,2,4]triazolo[1,5-c]pyridin-2-]cyclopropyl carboxamide (8 mg, 2.7 yield). .sup.1H NMR (400 MHz, METHANOL-d.sub.4) =9.99 (s, 1H), 9.57-9.44 (m, 1H), 8.52-8.44 (m, 1H), 8.39 (d, J=7.3 Hz, 1H), 8.33 (d, J=8.3 Hz, 1H), 8.01-7.68 (m, 1H), 5.57 (d, J=8.8 Hz, 2H), 5.36 (d, J=9.0 Hz, 2H), 4.47 (s, 2H), 1.81-1.61 (m, 4H). MS (ESI) Calcd. for C.sub.19H.sub.18F.sub.2N.sub.8O.sub.3S [M+H].sup.+ 477, Found 477.

Example 13

(51) ##STR00043##

Step 1: Preparation of N-[5-[1-[3-(cyanomethyl)-1-methylsulfony-azetidine-3-]pyrazole-4-]-[1,2,4]triazolo[1,5-a]pyrimidin2-]cyclopropyl carboxamide

(52) Intermediate 8 (150 mg, 314.9 umol, trifluoroacetate) was suspended in dichloromethane (2 mL). DIEA (203 mg, 1.6 mmol), DMAP (11 mg, 94.5 mmol) and MsCl (180 mg, 1.6 mmol) was added in order. The reaction mixture was stirred at 15 C. for reacting for 12 hours. LC-MS showed that the raw material was completely reacted, and the target product was detected. The reaction mixture was concentrated under reduced pressure to remove DCM. The residue was diluted with MeOH to a solution (5 mL), and separated through preparative HPLC (alkalinity) to give N-[5-[1-[3-(cyanomethyl)-1-methylsulfony-azetidine-3-]pyrazole-4-]-[1,2,4]triazolo[1,5-a]pyrimidin2-]cyclopropyl carboxamide (10 mg, 7.15% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) =11.17 (br. s., 1H), 9.27 (s, 1H), 8.79 (s, 1H), 7.83-7.68 (m, 1H), 7.62 (dd, J=7.8, 17.6 Hz, 2H), 4.51 (d, J=9.0 Hz, 2H), 4.34 (d, J=9.0 Hz, 2H), 3.69 (s, 2H), 3.16 (s, 3H), 2.25-2.00 (m, 1H), 0.95-0.80 (m, 4H). MS (ESI) Calcd. for C.sub.19H.sub.20N.sub.8O.sub.3S [M+H].sup.+ 441, Found 441.

Example 14

(53) ##STR00044##

Step 1: Preparation of N-(5-(1-(3-(cyanomethyl)-1-(ethylsulfony)azetidine-3-yl)-1H-pyrazole-4-yl]-[1,2,4]triazolo[1,5-a]pyridin-2-yl)cyclopropyl carboxamide(WX16)

(54) To a solution of Intermediate 6 (1.5 g, 5.6 mmol) and Intermediate 9 (1.5 g, 7.8 mmol) in acetonitrile (15 mL) was added dropwise DBU (1.7 g, 11.2 mmol) under the protection of nitrogen. The reaction mixture was stirred at 25 C. for 16 hours. TLC (petroleum ether/ethyl acetate=0:1) detected that the reaction was completed. The reaction mixture was poured into methanol (200 mL) at 0 C., a great number of solids were precipitated immediately, stirred for 10 min, and then filtered. The resulting solid was washed with methanol (5 mL) and dried under vacuum to give N-(5-(1-(3-(cyanomethyl)-1-(ethylsulfony)azetidine-3-yl)-1H-pyrazole-4-yl]-[1,2,4]triazolo[1,5-a]pyridin-2-yl)cyclopropyl carboxamide (1.60 g, 60% yield) as a product. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =9.25 (s, 1H), 8.79 (s, 1H), 7.76-7.70 (m, 1H), 7.64 (d, J=7.0 Hz, 1H), 7.60 (d, J=8.5 Hz, 1H), 4.49 (d, J=9.0 Hz, 2H), 4.32 (d, J=9.0 Hz, 2H), 3.68 (s, 2H), 1.26 (t, J=7.3 Hz, 3H), 0.91-0.85 (m, 4H). MS (ESI) Calcd. for C.sub.20H.sub.22N.sub.8O.sub.3S [M+H].sup.+ 455, Found 455.

Example 15

(55) ##STR00045##

Step 1: Preparation of tert-butyl 3-[4-(2-amino-[1,2,4]triazolo[1,5-a]pyridin-1-]-3-(cyanoethyl) azetidine-1-formate

(56) To 5-(1H-pyrazole-4-)-[1,2,4]triazolo[1,5-a]pyridin-2-amine (700 mg, 3.5 mmol) and tert-butyl 3-(cyanomethyl) azetidine-1-formate (747 mg, 3.9 mmol), dissolved in acetonitrile (20.00 mL), was added DBU (1.6 g, 10.5 mmol). The mixture was reacted at 40 C. for 3 hour. LC-MS showed that the raw material was completely reacted, and the target product was detected. The reaction mixture was poured into water (30 mL), stirred for 30 min. The water phase was extracted with ethyl acetate (20 mL3). The organic phase was combined, washed with saturated salt water (20 mL2), dried with anhydrous sodium sulfate, filtered, and distilled under reduced pressure to give tert-butyl 3-[4-(2-amino-[1,2,4]triazolo[1,5-a]pyridin-1-]-3-(cyanoethyl) azetidine-1-formate (1.33 g, crude product) as a brown solid. MS (ESI) Calcd. for C.sub.19H.sub.22N.sub.8O.sub.2 [M+H].sup.+ 395, Found 395.

Step 2: Preparation of tert-butyl 3-(cyanomethyl-3-[4-([1,2,4]triazolo[1,5-a]pyridin-5-) pyrazole-1-]azetidine-1-formate

(57) To 3-[4-(2-amino-[1,2,4]triazolo[1,5-a]pyridin-1-]-3-(cyanoethyl)azetidine-1-formate (150 mg, 380.3 umol) dissolved in tetrahydrofuran (2 mL) was added t-BuONO (59 mg, 570.5 umol), and stirred at 15 C. for 3 hours. LC-MS showed that the raw material was completely reacted, and the target product was detected. The reaction mixture was concentrated under reduced pressure, diluted with DCM (4 mL), and separated through preparative TLC (DCM:MeOH=10:1) to give tert-butyl 3-(cyanomethyl-3-[4-([1,2,4]triazolo[1,5-a]pyridin-5-)pyrazole-1-]azetidine-1-formate (80 mg, 55.4% yield). MS (ESI) Calcd. for C.sub.19H.sub.21N.sub.7O.sub.2 [M+H].sup.+ 380, Found 380.

Step 3: Preparation of 2-[3-[4-([1,2,4]triazolo[1,5-a]pyridin-5-)pyrazole-1-] azetidine-3-]acetonitrile

(58) Tert-butyl 3-(cyanomethy-3-[4-([1,2,4]triazolo[1,5-a]pyridin-5-)pyrazole-1-]azetidine-1-formate (80 mg, 210.9 umol) was suspended in DCM (1.5 mL). Trifluoroacetic acid (857 mg, 7.5 mmol) was added, and stirred at 15 C. for 3 hours. LC-MS showed that the reaction was completed, and the target product MS was detected. The reaction mixture was concentrated under reduced pressure to remove solvent and the rest of trifluoroacetic acid to 2-[3-[4-([1,2,4]triazolo[1,5-a]pyridin-5-) pyrazole-1-] azetidine-3-]acetonitrile (129 mg, crude product) as brown dope. MS (ESI) Calcd. for C.sub.14H.sub.13N.sub.7 [M+H].sup.+ 280, Found 280.

Step 4: Preparation of 2-[3-[4-([1,2,4]triazolo[1,5-a]pyridin-5-)pyrazole-1-]-1-(trifluoromethylsulfonyl)azetidine-3-]acetonitrile

(59) 2-[3-[4-([1,2,4]triazolo[1,5-a]pyridin-5-) pyrazole-1-] azetidine-3-]acetonitrile (60 mg, 214.8 umol) was dissolved in DCM (2 mL), DMAP (13 mg, 107.4 umol) and Et.sub.3N (109 mg, 1.1 mmol) were added, then trifluoromethanesulfonyl chloride (47 mg, 279.3 umol) was add dropwise at 15 C. The reaction mixture was stirred at 15 C. for reacting for 4 hours. LC-MS showed that the raw material was completely reacted, and the target product MS was detected. The reaction mixture was concentrated under reduced pressure to remove solvent and give 2-[3-[4-([1,2,4]triazolo[1,5-a]pyridin-5-) pyrazole-1-]-1-(trifluoromethylsulfonyl)azetidine-3-]acetonitrile (25 mg, 28.3% yield) through preparative HPLC (alkalinity). .sup.1H NMR (400 MHz, DMSO-d.sub.6) =9.28 (br. s., 1H), 8.73 (d, J=17.8 Hz, 2H), 7.82 (br. s., 3H), 5.22-4.50 (m, 4H), 3.86 (br. s., 2H). MS (ESI) Calcd. for C.sub.15H.sub.12F.sub.3N.sub.7O.sub.2S [M+H].sup.+ 412, Found 412.

Example 16

(60) ##STR00046##

Step 1: Preparation of 2-(3-(4-(2-amino-[1,2,4]triazolo[1,5-a]pyridinyl)-1H-pyrazolyl)-1-(trifluoromethyl sulfonyl)cyclobutane)acetonitrile(WX18)

(61) 2-(3-(4-(2-amino-[1,2,4]triazolo[1,5-a]pyridinyl)-1H-pyrazolyl)cyclobutane-3) acetonitrile (200 mg, 489.8 umol) was dissolved in DCM (10 mL), and TEA (198 mg, 2 mmol) was added. The resulting mixture was cooled to 0 C., then trifluoromethanesulfonyl chloride (107 mg, 636 umol) was slowly added dropwise. After dropwise adding, the reaction was warmed to room temperature and reaction was carried out at room temperature for 12 hours. LC-MS showed that the reaction was completed. The solvent was spun dry under reduced pressure. The residue was dissolved with DMF, further purified and freeze-dried through preparative HPLC (HCl) to give 2-(3-(4-(2-amino-[1,2,4]triazolo[1,5-a]pyridinyl)-1H-pyrazolyl)-1-(trifluoromethylsulfonyl)cyclobutane)acetonitrile. .sup.1H-NMR (400 MHz, MeOD-d.sub.4) =9.14 (s, 1H), 8.56 (s, 1H), 7.58 (t, J=7.8, 1H), 7.41 (d, J=7.5 Hz, 1H), 7.30 (d, J=8.8 Hz, 1H), 4.98 (d, J=9.3 Hz, 2H), 4.68 (d, J=9.0 Hz, 2H), 3.67 (s, 2H). MS (ESI) Calcd. for C.sub.15H.sub.13F.sub.3N.sub.8O.sub.2S [M+H].sup.+ 427, Found 427.

(62) Preparation of WX19: WX19 was prepared in the same method as the preparation of WX18 (Step 1). It was purified and freeze-dried through preparative HPLC (HCl) to give 2-(3-(4-(2-amino-[1,2,4]triazolo[1,5-a]pyridinyl)-1H-pyrazolyl)-1-(methylsulfonyl) cyclobutane)acetonitrile. .sup.1H-NMR (400 MHz, MeOD-d.sub.4) =9.13 (s, 1H), 8.53 (s, 1H), 7.57 (m, 1H), 7.41 (d, J=6.8 Hz, 1H), 7.30 (d, J=8.5 Hz, 1H), 4.64 (d, J=9.3 Hz, 2H), 4.35 (d, J=9.3 Hz, 2H), 3.62 (s, 2H), 3.08 (s, 3H). MS (ESI) Calcd. for C.sub.15H.sub.16N.sub.8O.sub.2S[M+H].sup.+ 373, Found 373.

Example 17

(63) ##STR00047##

Step 1: Preparation of N-(8-bromo-[1,2,4]triazolo[1,5-a]pyridin-2-yl) cyclopropanecarboxamide

(64) To 8-bromo-[1,2,4]triazolo[1,5-a]pyridin-2-amine (1.0 g, 4.7 mmol) and triethylamine (1.4 g, 14.1 mmol), dissolved in acetonitrile (15.0 mL), was added dropwise cyclopropanecarboxylic acid chloride (1.5 g, 14.1 mmol). Then the mixture is stirred at 26 C. for reacting for 12 hours. LC-MC showed that the reaction was completed. The reaction mixture was distilled under reduced pressure to remove acetonitrile. The residue was added to H.sub.2O (5 mL) and water layer was extracted with DCM (15 mL3). The organic phase was combined, washed with saturated salt water (15 mL), dried with anhydrous sodium sulfate, filtered, and distilled under reduced pressure to remove the filtrate. The residue was purified through silica gel column chromatography (DCM/MeOH=20/1) to give a yellow solid (700 mg, 47.8% yield). MS (ESI) Calcd. for C.sub.10H.sub.9N.sub.4OBr [M+H].sup.+ 282, Found 282.

Step 2: Preparation of N-[8-(1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl) cyclopropanecarboxamide

(65) To N-(8-bromo-[1,2,4]triazolo[1,5-a]pyridin-2-yl) cyclopropanecarboxamide (700 mg, 2.5 mmol) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (579 mg, 3.0 mmol), dissolved in dioxane (25 mL) and water (6 mL), was respectively added potassium carbonate (1.0 g, 7.5 mmol) and Pd(dppf)Cl.sub.2 (182 mg, 249 umol). The system is vacuumed and filled with nitrogen. Then the mixture was heated to reflux for 1 hour. LC-MC showed that the reaction was completed. The reaction mixture was distilled under reduced pressure to remove the solvent. The residue was dissolved in DCM (50 mL) and water (10 mL). The organic layer was separated and the water layer was extracted with DCM (250 mL) twice. The organic phase was combined, washed with saturated salt water (10 mL), dried with anhydrous sodium sulfate, filtered, and distilled under reduced pressure to remove the filtrate. The residue was purified through silica gel column chromatography (EA/PE=3/1 to 1/1) to give a yellow solid (300 mg, 40.4% yield). MS (ESI) Calcd. for C.sub.13H.sub.12N.sub.60 [M+H].sup.+ 269, Found 269.

Step 3: Preparation of compound of N-(8-(1-(3-(cyanomethyl)-1-(ethylsulfonyl) azetidine-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl) cyclopropanecarboxamide(WX20)

(66) To N-[8-(1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl) cyclopropanecarboxamide (100 mg, 372.8 umol) and 2-(1-ethylsulfonyl azetidine-3-yl) acetonitrile (83 mg, 447.3 umol), dissolved in acetonitrile (15 mL), was added dropwise DBU (68 mg, 447.3 umol). The formed mixture was stirred at 26 C. for reacting for 12 hours. After TLC showed that the reaction was completed, the reaction mixture was distilled under reduced pressure to remove the solvent. The residue was dissolved in DCM (15 mL) and water (10 mL). The organic layer was separated and the water layer was extracted with DCM (15 mL2) twice. The organic phase was combined, washed with saturated salt water (10 mL), dried with anhydrous sodium sulfate, filtered, and distilled under reduced pressure to remove the filtrate. The residue was purified through preparative HPLC (alkaline method) to give (WX20)(65 mg, 37.98% yield): .sup.1H NMR (400 MHz, METHANOL-d.sub.4) 6=8.72 (s, 1H), 8.37 (d, J=6.8 Hz, 1H), 8.28 (s, 1H), 7.71 (d, J=7.3 Hz, 1H), 6.98 (t, J=7.0 Hz, 1H), 4.63 (d, J=9.0 Hz, 2H), 4.28 (d, J=9.0 Hz, 2H), 3.58 (s, 2H), 3.20 (q, J=7.3 Hz, 2H), 1.44-1.31 (m, 3H), 1.07 (quin, J=3.8 Hz, 2H), 0.96 (qd, J=3.7, 7.3 Hz, 2H). MS (ESI) Calcd. for C.sub.20H.sub.22N.sub.8O.sub.3S [M+H].sup.+ 455, Found 455.

(67) Preparation of WX21: WX21 was prepared in the same method as the preparation of WX20 (Step 3). .sup.1H NMR (400 MHz, CDCl.sub.3) =8.93-8.82 (m, 1H), 8.67 (s, 1H), 8.42 (d, J=6.3 Hz, 1H), 8.15 (s, 1H), 7.63 (d, J=7.3 Hz, 1H), 6.96 (t, J=7.2 Hz, 1H), 4.62 (d, J=9.3 Hz, 2H), 4.25 (d, J=9.3 Hz, 2H), 3.42 (s, 2H), 2.54-2.42 (m, 1H), 1.87 (br. s., 1H), 1.25-1.17 (m, 4H), 1.13-1.06 (m, 2H), 0.94 (dd, J=3.0, 7.5 Hz, 2H). MS (ESI) Calcd. for C.sub.21H.sub.22N.sub.8O.sub.3S [M+H].sup.+467, Found 467.

Example 18

(68) ##STR00048##

Step 1: Preparation of N-(8-bromo-[1,2,4]triazolo[1,5-a]pyridin-2-yl)-2,2,2-trifluoro-acetamide

(69) To a solution of 8-bromo-[1,2,4]triazolo[1,5-a]pyridin-2-amine (1.0 g, 4.7 mmol) and triethylamine (1.4 g, 14.1 mmol) dissolved in dichloromethane (25.00 mL), was added dropwise trifluoroacetic acid (3.0 g, 14.1 mmol). The formed reaction mixture was stirred at 26 C. for reacting for 12 hours. After LC-MS showed that the reaction was completed, the reaction mixture was distilled under reduced pressure to remove the solvent. The residue was dissolved in DCM (50 mL) and saturated water (10 mL). The organic layer was separated and the water layer was extracted with DCM (50 mL2) twice. The organic phase was combined, washed with saturated salt water (10 mL), dried with anhydrous sodium sulfate, filtered, and distilled under reduced pressure to remove the filtrate to give N-(8-bromo-[1,2,4]triazolo[1,5-a]pyridin-2-yl)-2,2,2-trifluoro-acetamide (1.1 g) as a crude product which was used directly without purification. MS (ESI) Calcd. for C.sub.8H.sub.4N.sub.4OBrF.sub.3 [M+H].sup.+ 310, Found 310.

Step 2: Preparation of 8-(1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-amine

(70) To N-(8-bromo-[1,2,4]triazolo[1,5-a]pyridin-2-yl)-2,2,2-trifluoro-acetamide (1.1 g, 3.6 mmol) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (1.0 g, 356 umol), dissolved in dioxane (25 mL) and water (6 mL), was respectively added potassium carbonate (492 mg, 3.6 mmol) and Pd(dppf)Cl.sub.2 (260 mg, 356 umol). The system is vacuumed and filled with nitrogen. Then the mixture was heated to reflux for 1 hour. After LC-MC showed that the reaction was completed, the reaction mixture was filtered, and the filtrate was washed with water (10 mL) and then extracted with EA (30 mL3). The organic phase was combined, washed with saturated salt water (10 mL), dried with anhydrous sodium sulfate, filtered, and distilled under reduced pressure to remove the filtrate. The residue was purified through silica gel column chromatography (DCM/MeOH=DCM to 20/1) to give (430 mg, 57.3% yield) as a yellow solid. MS (ESI) Calcd. for C.sub.9H.sub.8N.sub.6 [M+H].sup.+ 201, Found 201.

Step 3: Preparation of 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-a]pyridine-8-yl)pyrazole-1-yl]-1-ethyl sulfonyl-azetidine-3-yl] acetonitrile

(71) To 8-(1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-amine (50 mg, 249.7 umol) and 2-(1-ethylsulfonyl-azetidine-3-ylene) acetonitrile (56 mg, 299.7 umol), dissolved in acetonitrile (8 mL), was added dropwise DBU (46 mg, 299.7 umol). The reaction mixture was stirred at 26 C. for reacting for 12 hours. After LC-MS showed that the reaction was completed, the reaction mixture was distilled under reduced pressure to remove acetonitrile. To the residue was added water (10 mL), and then extracted with EA (10 mL3). The organic phase was combined, washed with saturated salt water (10 mL), dried with anhydrous sodium sulfate, filtered, and distilled under reduced pressure to remove the filtrate. The residue was purified through silica gel column chromatography (DCM/MeOH=20/1) to give 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-a]pyridine-8-yl)pyrazole-1-yl]-1-ethylsulfonylazetidine-3-yl]acetonitrile (50 mg, 49.22% yield) as a white solid. MS (ESI) Calcd. for C.sub.16H.sub.18N.sub.8SO.sub.2[M+H].sup.+ 387, Found 387.

Step 4: Preparation of N-(8-(1-(3-(cyanomethyl)-1-(ethylsulfonyl)azetidine-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)-2,2,2-trifluoroacetamide(WX22)

(72) To a solution of 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-a]pyridine-8-yl)pyrazole-1-yl]-1-ethylsulfonyl-azetidine-3-yl]acetonitrile (50 mg, 129.4 umol) and triethylamine (39.28 g, 388.2 umol) dissolved in dichloromethane (5 mL), was added dropwise trifluoroacetic anhydride (81.5 mg, 388.2 umol). The reaction mixture was stirred at 26 C. for reacting for 12 hours. After TLC showed that the reaction was completed, H.sub.2O (5 mL) was added and the organic layer was separated, the water layer was extracted with DCM (15 mL3) twice. The organic phase was combined, washed with saturated salt water (10 mL), and washed with saturated salt water (10 mL), dried with anhydrous sodium sulfate, filtered, and distilled under reduced pressure to remove the filtrate. The residue was purified through preparative thin layer chromatography (DMC:MeOH=20:1) to give (WX22)(29 mg, 46.5% yield). .sup.1H NMR (400 MHz, METHANOL-d.sub.4) =8.93-8.89 (m, 1H), 8.63-8.59 (m, 1H), 8.48 (s, 1H), 8.02 (dd, J=1.0, 7.3 Hz, 1H), 7.28-7.21 (m, 1H), 4.68-4.58 (m, 4H), 4.32 (s, 2H), 3.60 (s, 2H), 3.19 (q, J=7.4 Hz, 2H), 1.38 (t, J=7.3 Hz, 3H). MS (ESI) Calcd. for C.sub.18H.sub.17N.sub.8SO.sub.3F.sub.3[M+H].sup.+483, Found 483.

Example 19

(73) ##STR00049##

Step 1: Preparation of 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-a]pyridine-8-yl)pyrazole-1-yl]-1-cyclopropylsulfonyl-azetidine-3-yl]acetonitrile

(74) To 8-(1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-amine (250 mg, 1.3 mmol) and 2-(1-cyclopropylsulfonyl-azetidine-3-ylene)acetonitrile (297 mg, 1.5 mmol), dissolved in acetonitrile (25.00 mL), was added dropwise DBU (228 mg, 1.5 mmol). The reaction mixture was stirred at 26 C. for reacting for 12 hours. After LC-MS showed that the reaction was completed, H.sub.2O (5 mL) was added and the organic layer was separated, the water layer was extracted with DCM (215 mL) twice. The organic phase was combined, washed with saturated salt water (15 mL), dried with anhydrous sodium sulfate, filtered, and distilled under reduced pressure to remove the filtrate. The residue was purified through silica gel column chromatography (DCM/MeOH=20/1) to give 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-a]pyridine-8-yl) pyrazole-1-yl]-1-cyclopropylsulfonyl-azetidine-3-yl]acetonitrile (250 mg, 45.2% yield) as a yellow solid. MS (ESI) Calcd. for C.sub.17H.sub.18N.sub.8SO.sub.2[M+H].sup.+ 399, Found 399.

Step 2. Preparation of WX23

(75) To 2-[3-[4-(2-amino-[1,2,4]triazolo[1,5-a]pyridine-8-yl)pyrazole-1-yl]-1-cyclopropylsulfonyl-azetidine-3-yl]acetonitrile (50 mg, 125.5 umol) and triethylamine (38 g, 376.5 umol) dissolved in DCM (5 mL), was added dropwise trifluoroacetic anhydride (79 mg, 376.5 umol). The reaction mixture was stirred at 26 C. for reacting for 12 hours. After LC-MS showed that the reaction was completed, H.sub.2O (5 mL) was added and the organic layer was separated, the water layer was extracted with DCM (215 mL) twice. The organic phase was combined, washed with saturated salt water (15 mL), dried with anhydrous sodium sulfate, filtered, and distilled under reduced pressure to remove the filtrate. The residue was purified through preparative HPLC (alkalinity, 0-60) to give (WX23)(17 mg, 27.4% yield). .sup.1H NMR .sup.1H NMR (400 MHz, CDCl.sub.3) =9.09-9.04 (m, 1H), 8.74 (s, 1H), 8.55-8.52 (m, 1H), 8.19 (s, 1H), 7.79-7.74 (m, 1H), 7.12 (t, J=7.0 Hz, 1H), 4.64 (d, J=9.3 Hz, 2H), 4.27 (d, J=9.3 Hz, 2H), 3.44 (s, 2H), 2.50-2.43 (m, 1H), 1.25-1.21 (m, 2H), 1.15-1.08 (m, 2H). MS (ESI) Calcd. for Cl.sub.9H.sub.17N.sub.8SO.sub.3 [M+H].sup.+ 495, Found 495.

Example 20

(76) ##STR00050##

Step 1: Preparation of tert-butyl 3-(cyanomethyl)-3-(4-(2-(2-methoxyacetamide)-[1,2,4]triazolo[1,5-c]pyridin-5-yl]-1H-pyrazole-1-yl)azetidine-1-carboxylate (1)

(77) To Intermediate 2 (0.1 g, 0.25 mmol) and triethylamine (0.15 mL, 1.2 mmol) dissolved in DMF (10.00 mL) was added 2-methoxyacetyl chloride (65 mg, 0.5 mmol). The resulting mixture was stirred at 60 C. for reacting for 16 hours until LC-MS showed that the reaction was completed. The mixture was poured into 10 ml of water and extracted with ethyl acetate (10 ml3). The organic phase was combined, washed with saturated salt water (20 mL), dried with anhydrous sodium sulfate, and concentrated to give a crude product (120 mg) which was directly used in the next step. MS (ESI) Calcd. for C.sub.21H.sub.25N.sub.9O.sub.4[M+H.sup.+] 468, Found 468.

Step 2: Preparation of N-(5-(1-(3-(cyanomethyl) azetidine-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-c]pyridin-2-yl)-2-methoxyacetamide (2)

(78) Tert-butyl 3-(cyanomethyl)-3-(4-(2-(2-methoxyacetamide)-[1,2,4]triazolo[1,5-c]pyridin-5-yl]-1H-pyrazole-1-yl)azetidine-1-carboxylate (100 mg, 0.2 mmol) was dissolved in dichloromethane (5 mL), and then TFA (5 ml) was added. The resulting mixture was stirred at 10 C. for reacting for 1 hour. LC-MS showed that the reaction was completed and the solvent was concentrated to give 100 mg of crude product which was directly used in the next step. MS (ESI) Calcd. for C.sub.16H.sub.17N.sub.9O.sub.2[M+H].sup.+ 482, Found 482.

Step 3: Preparation of N-(5-(1-(3-(cyanomethyl)-1-((trifluoromethyl)sulfonyl) azetidine-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-c]pyridin-2-yl)-2-methoxyacetamide (WX24)

(79) N-(5-(1-(3-(cyanomethyl)azetidine-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-c]pyridin-2-yl)-2-methoxyacetamide (50 mg, 0.14 mmol) was dissolved in dichloromethane (5 mL), triethylamine (42 mg, 0.4 mmol) and then trifluoromethanesulfonyl chloride (47 mg, 0.28 mmol) was added. The resulting mixture was stirred at 10 C. for reacting 1 hour. LC-MS showed that the reaction was completed and the solvent was concentrated to give a crude product (50 mg). The crude product was separated through preparative HPLC (alkalinity) to give N-(5-(1-(3-(cyanomethyl)-1-((trifluoromethyl)sulfonyl)azetidine-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-c]pyridin-2-yl)-2-methoxyacetamide(WX24, 10 mg) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.30 (s, 1H), 8.89 (s, 1H), 8.36 (d, J=6.27 Hz, 1H), 7.66 (d, J=6.02 Hz, 1H), 4.74 (s, 2H), 3.86 (s, 2H), 3.40 (s, 4H). MS (ESI) Calcd. for C.sub.17H.sub.16F.sub.3N.sub.9O.sub.4S [M+H].sup.+ 495, Found 495.

(80) Preparation of WX25: N-(5-(1-(3-(cyanomethyl)-1-((methylsulfonyl)azetidine-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-c]pyridin-2-yl)-2-methoxyacetamide(WX25) was prepared in the same method as the preparation of WX24 (Step 3). .sup.1H NMR (400 MHz, CDCl.sub.3) =9.29-9.32 (m, 1H), 8.67-8.71 (m, 1H), 8.28-8.33 (m, 1H), 7.44-7.48 (m, 1H), 4.60-4.68 (m, 1H), 4.30-4.36 (m, 2H), 4.13-4.19 (m, 1H), 3.58 (s, 2H), 3.41-3.48 (m, 1H), 3.04 (s, 3H), 1.45 (s, 3H). MS (ESI) Calcd. for C.sub.17H.sub.19N.sub.9O.sub.4S [M+H].sup.+ 446, Found 446.

Example 21

(81) ##STR00051##

Step 1: Preparation of tert-butyl 3-(4-(2-(2-cyanoacetamide)-[1,2,4]triazolo[1,5-c]pyridin-5-yl)-1H-pyrazole-1-yl)-3-(cyanomethyl)azetidine-1-carboxylate (1)

(82) To Intermediate 2 (0.1 g, 0.3 mmol) and triethylamine (0.17 ml, 1.3 mmol), dissolved in DMF (10 mL), was added 2-cyanoacetyl chloride (131 mg, 1.3 mmol). The resulting mixture was stirred at 60 C. for reacting for 2 hours until LC-MS showed that the reaction was completed. The mixture was poured into water (10 mL) and extracted with ethyl acetate (10 mL3). The organic phase was combined, dried with anhydrous sodium sulfate, and concentrated to give tert-butyl 3-(4-(2-(2-cyanoacetamide)-[1,2,4]triazolo[1,5-c]pyridin-5-yl)-1H-pyrazole-1-yl)-3-(cyanomethyl)azetidine-1-carboxylate (100 mg, crude product) which was directly used in the next step. MS (ESI) Calcd. for C.sub.21H.sub.22N.sub.10O.sub.3[M+H].sup.+ 463, Found 463.

Step 2: Preparation of 2-cyano-N-(5-(1-(3-(cyanomethyl)azetidine-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-c]pyridin-2-yl)acetamide (2)

(83) Tert-butyl 3-(4-(2-(2-cyanoacetamide)-[1,2,4]triazolo[1,5-c]pyridin-5-yl)-1H-pyrazole-1-yl)-3-(cyanomethyl)azetidine-1-carboxylate (1)(100 mg, 0.2 mmol) was dissolved in dichloromethane (5 mL) and TFA (5 mL) was added. The resulting mixture was stirred at 10 C. for reacting for 1 hour, LC-MS showed that the reaction was completed. The solvent was concentrated to give 2-cyano-N-(5-(1-(3-(cyanomethyl)azetidine-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-c]pyridin-2-yl)acetamide (2)(100 mg, crude product) which was directly used in the next step. MS (ESI) Calcd. for C.sub.16H.sub.14N.sub.10O[M+H].sup.+ 463, Found 463.

Step 3: Preparation of 2-cyano-N-(5-(1-(3-(cyanomethyl)-1-(methanesulfonamide) azetidine-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-c]pyridin-2-yl)acetamide (WX26)

(84) 2-cyano-N-(5-(1-(3-(cyanomethyl)azetidine-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-c]pyridin-2-yl)acetamide (50 mg, 0.14 mmol) was dissolved in dichloromethane (5 mL), triethylamine (42 mg, 0.4 mmol) and methanesulfonyl chloride (47 mg, 0.28 mmol). The resulting mixture was stirred at 10 C. for reacting for 1 hour, LC-MS showed that the reaction was completed. The solvent was concentrated to give 50 mg of crude product. The crude product was separated through preparative HPLC (alkalinity) to give 2-cyano-N-(5-(1-(3-(cyanomethyl)-1-(methanesulfonamide) azetidine-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-c]pyridin-2-yl)acetamide(WX26 10 mg). .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.27 (s, 1H), 8.83 (s, 1H), 8.25-8.52 (m, 1H), 7.67 (d, J=6.27 Hz, 1H), 4.56 (d, J=9.29 Hz, 4H), 4.34 (d, J=9.29 Hz, 4H), 3.73 (s, 3H).sub. MS (ESI) Calcd. for C.sub.17H.sub.16N.sub.10O.sub.3S[M+H].sup.+ 441, Found 441.

Example 22

Step 1: Preparation of 1-(3-bromophenyl) thiourea(2)

(85) ##STR00052## ##STR00053##

(86) To a dilute hydrochloric acid solution (1M, 50 mL) of 3-bromaniline (30.0 g, 174 mmol) was added potassium thiocyanate (20.0 g, 205.8 mmol) at room temperature. The mixture was stirred at 100 C. for reacting for 12 hours. TLC (PE:EA=1:1) detected that there was part of 3-bromaniline left (about 20%). The reaction mixture was cooled to 0 C., then was alkalified with ammonium hydroxide to pH=10. The resulting violet emulsion was stirred continuously for half an hour and then extracted with ethyl acetate (200 mL4). The organic phase was combined, washed with saturated salt water (30 mL), dried with anhydrous sodium sulfate, and filtered and spun dry to give thick violet suspension. Until a little cool, dichloromethane (50 mL) was added and then cooled to 0 C. in ice bath. After suction filtration of insoluble lavender solid, washed with a small amount of dichloromethane (10 mL2), and dried under vacuum to give 1-(3-bromophenyl) thiourea (25 g, 55.8% yield). .sup.1H NMR (400 MHz, CDCl3) 07.46 (s, 1H), 7.39-7.44 (m, 1H), 7.28-7.32 (m, 1H), 7.25 (s, 1H). MS (ESI) Calcd. for C.sub.7H.sub.7BrN.sub.2S [M+H].sup.+ 230, Found 230.

Step 2: Preparation of 7-bromobenzo[d]thiazole-2-amine(3)

(87) To 1-(3-bromophenyl) thiourea (5.0 g, 21.6 mmol) in acetic acid (50 mL) was added dropwise a solution of liquid bromine (4.7 g, 29.2 mmol) in chloroform (5 mL) at 0 C. The mixture was stirred at 85 C. for reacting for 3 hours. TLC (PE:EA=1:1) showed that raw material was completely reacted, and two points were obtained. The reaction mixture was filtered while it was hot. The insoluble solid was washed with a small amount of dichloromethane (10 mL2) and dried to give a product as yellow solid (4.2 g, 50% yield). The filtrate was spun dry to give yellow suspension, and the residue was beat with dichloromethane (20 mL). After suction filtration of the insoluble substance, washed with DCM (5 mL2), and dried under vacuum to give a product as a yellow solid (1.2 g, 13% yield). One part of the product P1 and P2 were dissolved in water (1 mL) respectively, alkalified with ammonium hydroxide to pH=10 and then extracted with ethyl acetate (0.2 mL). The two extracts were blew-dry with nitrogen, and were used directly for NMR detection. NMR showed that there was mainly a by-product of 5-bromide isomers in P1 and mainly 7-bromide product. The resulting crude product was directly used in the next step without further purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 07.13-7.16 (m, 1H), 7.16-7.22 (m, 2H), 7.32 (dd, J=7.28, 1.51 Hz, 1H), 7.48 (d, J=1.76 Hz, 1H), 7.62 (d, J=8.53 Hz, 1H), 7.71 (br. s., 1H), 7.73 (s, 2H). MS (ESI) Calcd. for C.sub.7H.sub.5BrN.sub.2S[M+H].sup.+ 228, Found 228.

Step 3: Preparation of N-(7-bromobenzo[d]thiazole-2-yl)cyclopropyl carboxamide (4)

(88) To 7-bromobenzo[d]thiazole-2-amine (1.2 g, 3.9 mmol, HBr salt) and triethylamine (1.6 g, 15.5 mmol) in acetonitrile (50 mL) was added dropwise cyclopropanecarbonyl chloride (1.2 g, 11.6 mmol) at 0 C. under the protection of nitrogen. The mixture was stirred at 30 C. for reacting for 12 hours. TLC (PE:EA=1:1) detected and showed that the main point was the desired mono-substituted product. The reaction mixture was quenched with water (60 mL) and extracted with ethyl acetate (30 mL3). The organic phase was combined, washed with saturated salt water (5 mL), dried with anhydrous sodium sulfate, and filtered and spun dry. The residue was purified through column chromatography (PE:EA=5:1) to give N-(7-bromobenzo[d]thiazole-2-yl) cyclopropyl carboxamide (580 mg, 40% yield) as a yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =12.85 (s, 1H), 7.75 (d, J=8.03 Hz, 1H), 7.52 (d, J=7.78 Hz, 1H), 7.38-7.43 (m, 1H), 1.98-2.05 (m, 1H), 0.95-1.02 (m, 4H). MS (ESI) Calcd. for C.sub.11H.sub.9BrN.sub.2OS [M+H].sup.+ 296, Found 296.

Step 4: Preparation of N-(7-(1H-pyrazole-4-yl)benzo[d]thiazole-2-yl) cyclopropanecarboxamide (5)

(89) To N-(7-bromobenzo[d]thiazole-2-yl) cyclopropanecarboxamide (480 mg, 1.6 mmol) and 1H-pyrazole-4-boronic acid pinacol ester (317 mg, 1.6 mmol) in dioxane (15 mL) was added Pd(dppf)Cl2 (119 mg, 162 umol), K.sub.2CO.sub.3 (672 mg, 4.9 mmol) and H.sub.2O (2.5 mL) under the protection of nitrogen. The mixture was stirred at 90 C. for reacting for 12 hours. TLC (PE:EA=1:1) detected and showed a new point. LC-MS detected the target product. The reaction mixture was diluted with water (100 mL) and then extracted with ethyl acetate (30 mL3). The organic phase was combined, washed with saturated salt water (5 mL), dried with anhydrous sodium sulfate, and filtered and spun dry. The residue was purified through column chromatography (PE:EA=1:1) to give N-(7-(1H-pyrazole-4-yl)benzo[d]thiazole-2-yl) cyclopropane carboxamide (80 mg, 15.63% yield) as a yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =13.22 (br. s., 1H), 12.70 (s, 1H), 8.23 (s, 1H), 7.99 (s, 1H), 7.64 (d, J=8.03 Hz, 1H), 7.52-7.55 (m, 1H), 7.45-7.50 (m, 1H), 2.03 (t, J=4.52 Hz, 1H), 0.95-1.00 (m, 4H). MS (ESI) Calcd. for C.sub.14H.sub.12N.sub.4OS [M+H].sup.+ 285, Found 285.

Step 5: Preparation of 7-(1H-pyrazole-4-yl)benzo[d]thiazole-2-amine (6)

(90) To a solution of N-(7-(1H-pyrazole-4-yl)benzo[d]thiazole-2-yl)cyclopropyl carboxamide (120 mg, 422.1 ummol) in methanol (3 mL) was added dropwise NaOH (240 mg, 6 mmol) in an aqueous solution (1 mL). The mixture was stirred at 80 C. for reacting for 12 hours. LCMS detected and showed that the reaction was completed. The reaction mixture was diluted with water (50 mL), neutralized with 1M of HCl to pH=7, and extracted with ethyl acetate (15 mL4). The organic phase was combined, washed with saturated salt water (5 mL), dried with anhydrous sodium sulfate, and filtered and spun dry to give the residue of 7-(1H-pyrazole-4-yl)benzo[d] thiazole-2-amine (100 mg, 87.7% yield) as a crude product which was directly used in the next step without further purification. MS (ESI) Calcd. for C.sub.10H.sub.8N.sub.4S [M+H].sup.+ 216, Found 216.8.

Step 6: Preparation of tert-butyl 3-(4-(2-aminobenzo[d]thiazole-7-yl)-1H-pyrazole-1-yl)-3-(cyanomethyl)azetidine-1-carbonate (7)

(91) To a solution of 7-(1H-pyrazole-4-yl)benzo[d]thiazole-2-amine (100 mg, 462.4 umol) and tert-butyl 3-(cyanomethyl)azetidine-1-carbonate (90 mg, 463.4 umol) in acetonitrile (3 mL) was added dropwise DBU (140.8 mg, 924.8 umol) under the protection of nitrogen. The mixture was stirred at 30 C. for reacting for 12 hours. LC-MS detected and showed that the reaction was completed. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (20 mL3). The organic phase was combined, washed with saturated salt water (30 mL), dried with anhydrous sodium sulfate, and filtered and spun dry to give the residue of tert-butyl 3-(4-(2-aminobenzo[d]thiazole-7-yl)-1H-pyrazole-1-yl)-3-(cyanomethyl)azetidine-1-carbonate (190 mg, 80% yield) as a crude product which was directly used in the next step without further purification. MS (ESI) Calcd. for C.sub.20H.sub.22N.sub.6O.sub.2S[M+H].sup.+ 410, Found 411.

Step 7: Preparation of 2-(3-(4-(2-aminobenzo[d]thiazole-7-yl)-1H-pyrazole-1-yl)-azetidine-3-yl) methyl cyanide(8)

(92) Tht mixture of tert-butyl 3-(4-(2-aminobenzo[d]thiazole-7-yl)-1H-pyrazole-1-yl)-3-(cyanomethyl) azetidine-1-carbonate (180 mg, 438.5 umol) and hydrochloride ethyl acetate (30 mL) was stirred at 25 C. for reacting for 1 hour. LCMS detected that the reaction was completed. The reaction mixture was spun dry directly to give a yellow solid of 2-(3-(4-(2-aminobenzo[d]thiazole-7-yl)-1H-pyrazole-1-yl)-azetidine-3-yl) methyl cyanide (150 mg, 78.9% yield, HCl salt) as a crude product which was directly used in the next step without further purification. MS (ESI) Calcd. for C.sub.15H.sub.14N.sub.6S [M+H].sup.+ 310, Found 310.

Step 8: Preparation of 2-(3-(4-(2-aminobenzo[d]thiazole-7-yl)-1H-pyrazole-1-yl)-(methyl sulfonyl)azetidine-3-yl) methyl cyanide(WX27)

(93) To 2-(3-(4-(2-aminobenzo[d]thiazole-7-yl)-1H-pyrazole-1-yl)-azetidine-3-yl) methyl cyanide (150 mg, 345.9 umol, HCl salt) and Et.sub.3N (140 mg, 1.4 mmol) in dichloromethane (3 mL) was added dropwise MsCl (80 mg, 698.9 umol) at 0 C. under the protection of nitrogen. The mixture was stirred at 0 C. for reacting for 1 hour. LC-MS detected that the reaction was completed. The reaction mixture was quenched with water (50 mL) and extracted with dichloromethane (15 mL3). The organic phase was combined, washed with saturated salt water (5 mL), dried with anhydrous sodium sulfate, and filtered and spun dry. The residue was purified through preparative thin layer chromatography (DMC:MeOH=10:1) to give 2-(3-(4-(2-aminobenzo[d]thiazole-7-yl)-1H-pyrazole-1-yl)-(methylsulfonyl)azetidine-3-yl) methyl cyanide (30 mg, 20.1% yield) as a yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =8.55 (s, 1H), 8.07 (s, 1H), 7.56 (s, 2H), 7.28-7.31 (m, 2H), 7.25-7.28 (m, 1H), 4.54 (d, J=9.29 Hz, 2H), 4.27 (d, J=9.29 Hz, 2H), 3.66 (s, 2H), 3.14 (s, 3H). MS (ESI) Calcd. for C.sub.16H.sub.16N.sub.6O.sub.2S.sub.2[M+H].sup.+ 389, Found 389.

Example 23

(94) ##STR00054##

Step 1: Preparation of tert-butyl 3-(cyanomethyl)-3-(4-(2-(cyclopropanecarboxamido) benzo[d]thiazole-7-yl)-1H-pyrazole-1-yl) azetidin-1-carbonate

(95) To N-[7-(1H-pyrazole-4-yl)-1,3-benzothiazol-2-yl] cyclopropanecarboxamide (100 mg, 351.7 umol) and tert-butyl 3-(cyanomethylene) azetidin-carbonate (100 mg, 513.5 umol) in acetonitrile (3 mL) was added dropwise DBU (107 mg, 703.4 umol) under the protection of nitrogen. The mixture was stirred at 25 C. for reacting for 12 hours. TLC (PE:EA=1:1) detected and showed that the reaction was completed. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (20 mL3). The organic phase was combined, washed with saturated salt water (5 mL), dried with anhydrous sodium sulfate, and filtered and spun dry. The residue was separated and purified through preparative TLC (PE:EA=1:1) to give tert-butyl 3-(cyanomethyl)-3-(4-(2-(cyclopropanecarboxamido)benzo[d]thiazole-7-yl)-1H-pyrazole-1-yl) azetidin-1-carbonate (80 mg, 47.5% yield) as a yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) =11.30 (br. s., 1H), 8.05 (d, J=12.05 Hz, 2H), 7.75 (d, J=7.78 Hz, 1H), 7.48-7.53 (m, 1H), 7.43-7.47 (m, 1H), 4.58 (d, J=9.79 Hz, 2H), 4.32 (d, J=9.54 Hz, 2H), 3.31 (s, 2H), 1.74 (dq, J=7.97, 3.95 Hz, 1H), 1.51 (s, 9H), 1.27-1.31 (m, 2H), 1.01-1.09 (m, 2H). MS (ESI) Calcd. for C.sub.24H.sub.26N.sub.6O.sub.3S [M+H].sup.+ 479, Found 479.

Step 2: Preparation of N-(7-(1-(3-(cyanomethyl)azetidin-3-yl)-1H-pyrazole-4-yl) benzo[d]thiazole-2-yl) cyclopropanecarboxamide

(96) Tht mixture of tert-butyl 3-(cyanomethyl)-3-(4-(2-(cyclopropanecarboxamido) benzo[d]thiazole-7-yl)-1H-pyrazole-1-yl) azetidin-1-carbonate (80 mg, 167.2 umol) and hydrochloride ethyl acetate (30 mL) was stirred at 25 C. for reacting for 1 hour. LCMS detected that the reaction was completed. The reaction mixture was spun dry directly. The residue was dissolved in water (50 mL), adjusted with the aqueous solution of saturated NaHCO.sub.3 to weakly alkaline (pH>7), and then extracted with ethyl acetate (15 mL3). The organic phase was combined, washed with saturated salt water (5 mL), dried with anhydrous sodium sulfate, and filtered and spun dry to give a yellow solid of N-(7-(1-(3-(cyanomethyl)azetidin-3-yl)-1H-pyrazole-4-yl) benzo[d]thiazole-2-yl) cyclopropanecarboxamide (60 mg, 85.4% yield, purity of 90%) as a crude product which was directly used in the next step without further purification. MS (ESI) Calcd. for C.sub.19H.sub.18N.sub.60S[M+H].sup.+ 37, Found 378.

Step 3: Preparation of N-(7-(1-(3-(cyanomethyl)-1-(methylsulfonyl)azetidin-3-yl)-1H-pyrazole-4-yl)benzo[d]thiazole-2-yl) cyclopropanecarboxamide

(97) To a solution of N-(7-(1-(3-(cyanomethyl)azetidin-3-yl)-1H-pyrazole-4-yl) benzo[d]thiazole-2-yl) cyclopropanecarboxamide (60 mg, 158.54 umol) and triethylamine (50 mg, 494.6 umol) in dichloromethane (5 mL) was added dropwise methylsulfonyl chloride (50 mg, 436.5 umol). The mixture was stirred at 0 C. for 0.5 hour and stirred continuously at 25 C. for 1 hour. Both LC-MS and TLC detected and showed that the reaction was completed. The reaction mixture was quenched with water (50 mL) and extracted with ethyl acetate (15 mL3). The organic phase was combined, washed with saturated salt water (5 mL), dried with anhydrous sodium sulfate, and filtered and spun dry. The residue was separated and purified through preparative TLC (PE:EA=1:1) to give N-(7-(1-(3-(cyanomethyl)-1-(methylsulfonyl)azetidin-3-yl)-1H-pyrazole-4-yl)benzo[d]thiazole-2-yl) cyclopropanecarboxamide (45 mg, 62% yield) as a white solid.

(98) 1H NMR (400 MHz, CDCl.sub.3) =12.26 (br. s., 1H), 8.07 (s, 2H), 7.74 (d, J=7.78 Hz, 1H), 7.46-7.52 (m, 1H), 7.39-7.45 (m, 1H), 4.63 (d, J=9.03 Hz, 2H), 4.30 (d, J=9.03 Hz, 2H), 3.41 (s, 2H), 3.04 (s, 3H), 1.72-1.80 (m, 1H), 1.26 (d, J=3.26 Hz, 2H), 1.01 (dd, J=7.40, 2.89 Hz, 2H). MS (ESI) Calcd. for C.sub.20H.sub.20N.sub.6O.sub.3S.sub.2[M+H].sup.+ 457, Found 457.

Example 24

(99) ##STR00055## ##STR00056##

Step 1: Preparation of 2-(benzyloxy)-1-bromo-3-nitrobenzene

(100) To 2-bromo-6-nitrophenol (5.2 g, 23.9 mmol) and K.sub.2CO.sub.3 (3.6 g, 26.3 mmol) in acetonitrile (100 mL) was benzyl bromide (4.3 g, 25.3 mmol). The mixture was stirred at 100 C. for 3 hour. TLC (PE:EA=10:1) detected and showed that the reaction was completed. The reaction mixture was filtered and the solid was eluted with ethyl acetate (10 mL3). After the filtrate was spun dry, the residue was dissolved in ethyl acetate, and washed with water (20 m/L) and saturated salt water (20 mL). The organic phase was dried with anhydrous sodium sulfate, filtered and spun dry to give a yellow solid of 2-(benzyloxy)-1-bromo-3-nitrobenzene (7.50 g, 91.51% yield) as a crude product which was directly used in the next step without further purification. .sup.1H NMR (400 MHz, CDCL.sub.3) 7.84 (d, J=8.03 Hz, 1H), 7.79 (d, J=8.03 Hz, 1H), 7.56 (d, J=6.53 Hz, 2H), 7.37-7.45 (m, 3H), 7.16 (t, J=8.28 Hz, 1H), 5.21 (s, 2H). MS (ESI) Calcd. for C.sub.13H.sub.10BrNO.sub.3 [M+H].sup.+ 308, Found 308.

Step 2: Preparation of 4-(2-(benzyloxy)-3-nitrophenyl)-1H-pyrazole

(101) To 2-(benzyloxy)-1-bromo-3-nitrobenzene (1.0 g, 3.3 mmol) and ter-butyl 1H-pyrazole-4-boronic acid pinacol ester-1-carbonate (1.0 g, 3.4 mmol) in dioxane (30 mL) was added Pd(dppf)Cl.sub.2 (250 mg, 341.7 umol), K.sub.2CO.sub.3 (1.4 g, 9.8 mmol) and H.sub.2O (2.5 mL) under the protection of nitrogen. The mixture was stirred at 100 C. for reacting for 12 hours. Both TLC (PE:EA=1:1) and LC-MS detected and showed that the reaction was completed. The reaction mixture was diluted with water (20 mL), and extracted with ethyl acetate (20 mL3). The organic phase was combined, washed with saturated salt water (5 mL), dried with anhydrous sodium sulfate, and filtered and spun dry. The residue was separated and purified through column chromatography (PE:EA=1:1) to give 4-(2-(benzyloxy)-3-nitrophenyl)-1H-pyrazole (900 mg, 89.1% yield) as a yellow oil product. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.03 (s, 2H), 7.70-7.78 (m, 2H), 7.33-7.39 (m, 5H), 7.28-7.32 (m, 1H), 4.87 (s, 2H). MS (ESI) Calcd. for C.sub.16H.sub.13N.sub.3O.sub.3 [M+H].sup.+ 296, Found 296.

Step 3: Preparation of ter-butyl 3-(4-(2-(benzyloxy)-3-nitrophenyl)-1H-pyrazole-1-yl)-3-(cyanomethyl) azetidin-1-carbonate

(102) To 4-(2-(benzyloxy)-3-nitrophenyl)-1H-pyrazole (900 mg, 3.1 mmol) and ter-butyl 3-(cyanomethylene) azetidin-1-carbonate (900 mg, 4.6 mmol) in acetonitrile (20 mL) was added dropwise DBU (928 mg, 6.1 mmol) at 0 C. under the protection of nitrogen. The mixture was stirred at 25 C. for reacting for 3 hours. TLC (PE:EA=1:1) detected and showed that the reaction was completed. The reaction mixture was quenched with water (60 mL) and extracted with ethyl acetate (20 mL3). The organic phase was combined, washed with saturated salt water (5 mL), dried with anhydrous sodium sulfate, and filtered and spun dry. The residue was separated and purified through column chromatography (PE:EA=1:1) to give ter-butyl 3-(4-(2-(benzyloxy)-3-nitrophenyl)-1H-pyrazole-1-yl)-3-(cyanomethyl) azetidin-1-carbonate (1.00 g, 60.3% yield) as a yellow oil product. .sup.1H NMR (400 MHz, CDCl.sub.3) =7.96 (d, J=9.03 Hz, 2H), 7.71-7.76 (m, 2H), 7.32-7.40 (m, 5H), 7.27-7.31 (m, 1H), 4.90 (s, 2H), 4.24 (d, J=9.54 Hz, 2H), 4.12 (d, J=9.03 Hz, 2H), 3.16 (s, 2H), 1.48 (s, 9H). MS (ESI) Calcd. for C.sub.26H.sub.27N.sub.5O.sub.5 [M+H].sup.+ 490, Found 490.

Step 4: Preparation of 2-(3-(4-2-(benzyloxy)-3-nitrophenyl-1H-pyrazole-1-yl) azetidin-3-yl)acetonitrile

(103) The mixture of 3-(4-(2-(benzyloxy)-3-nitrophenyl)-1H-pyrazole-1-yl)-3-(cyanomethyl)azetidin-1-carbonate (1.0 g, 2.04 mmol) and hydrochloride ethyl acetate (50 mL) was stirred at 25 C. for reacting for 3 hours. LC-MS detected and showed that the reaction was completed. After the reaction mixture was spun dry, the residue was dissolved in water (50 mL), adjusted with saturated NaHCO.sub.3 solution to neutrality pH=7, and then extracted with ethyl acetate (15 mL4). The organic phase was combined, washed with saturated salt water (2 mL), dried with anhydrous sodium sulfate, and filtered and spun dry to give a orange oil of 2-(3-(4-2-(benzyloxy)-3-nitrophenyl-1H-pyrazole-1-yl) azetidin-3-yl)acetonitrile (900 mg, 90.6% yield) as a crude product which can be directly used in the next step without further purification. MS (ESI) Calcd. for C.sub.21H.sub.19N.sub.5O.sub.3 [M+H].sup.+ 390, Found 390.

Step 5: Preparation of 2-(3-(4-2-(benzyloxy)-3-nitrophenyl-1H-pyrazole-1-yl)-1-(methylsulfonyl)azetidin-3-yl)acetonitrile

(104) To 2-(3-(4-2-(benzyloxy)-3-nitrophenyl-1H-pyrazole-1-yl) azetidin-3-yl)acetonitrile (900 mg, 2.3 mmol) and triethylamine (730 mg, 7.2 mmol) in dichloromethane (10 mL) was added dropwise methanesulfonyl chloride (529 mg, 4.6 mmol) at 0 C. under the protection of nitrogen. The mixture was stirred at 0 C. for reacting for 1 hour. LC-MS detected and showed that the reaction was completed. The reaction mixture was quenched with water (50 mL) and then extracted with ethyl acetate (20 mL3). The organic phase was combined, washed with saturated salt water (5 mL), dried with anhydrous sodium sulfate, and filtered and spun dry. The residue was separated and purified through column chromatography (PE:EA=1:1 (0.5% Et.sub.3N)) to give 2-(3-(4-2-(benzyloxy)-3-nitrophenyl-1H-pyrazole-1-yl)-1-(methyl sulfonyl)azetidin-3-yl)acetonitrile (450 mg, 37.5% yield) as a yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3) =7.98 (d, J=12.55 Hz, 2H), 7.71-7.78 (m, 2H), 7.34-7.41 (m, 5H), 7.30 (t, J=8.03 Hz, 1H), 4.88-4.93 (m, 2H), 4.33 (d, J=9.03 Hz, 2H), 4.09 (d, J=9.54 Hz, 2H), 3.21 (s, 2H), 2.95 (s, 3H). MS (ESI) Calcd. for C.sub.22H.sub.21N.sub.5O.sub.5S [M+H].sup.+ 468, Found 468.

Step 6: Preparation of 2-(3-(4-(3-amino-2-hydroxypheyl)-1H-pyrazole-1-yl)-1-(methylsulfonyl)azetidin-3-yl)acetonitrile

(105) To 2-(3-(4-2-(benzyloxy)-3-nitrophenyl-1H-pyrazole-1-yl)-1-(methylsulfonyl) azetidin-3-yl)acetonitrile (400 mg, 855.6 umol) in ethyl acetate (10 mL) was added Pd/C (200 mg, 1.9 mmol) under the protection of nitrogen. The mixture was stirred at 25 C. for reacting for 1.5 hours. LC-MS detected and showed that the reaction was completed. The reaction mixture was filtered to remove Pd/C, and concentrated by rotary evaporation at low temperature to give 2-(3-(4-(3-amino-2-hydroxypheyl)-1H-pyrazole-1-yl)-1-(methylsulfonyl)azetidin-3-yl)acetonitrile (300 mg, 80.75%) as a pale yellow, which can be directly used in the next step without further purification. MS (ESI) Calcd. for C.sub.15H.sub.17N.sub.5O.sub.3S [M+H].sup.+ 348, Found 348.

Step 7: Preparation of 2-(3-(4-(2-aminobenzo[d]oxazole-7-yl)-1H-pyrazole-1-yl)-1-(methylsulfonyl)azetidin-3-yl)acetonitrile

(106) To 2-(3-(4-(3-amino-2-hydroxypheyl)-1H-pyrazole-1-yl)-1-(methyl sulfonyl)azetidin-3-yl)acetonitrile (300 mg, 863.58 umol) in ethyl acetate (10 mL) solution was added cyanogen bromide (100 mg, 944.1 umol) under the protection of nitrogen. The mixture was stirred at 50 C. for reacting for 12 hours. LC-MS and TLC (DCM:MeOH=10:1) detected and showed that the reaction was completed. After the reaction mixture was directly spun dry, dissolved in DCM:MeOH=10:1 (3 mL), separated and purified through preparative TLC (DCM:MeOH=10:1) to give 2-(3-(4-(2-aminobenzo[d]oxazole-7-yl)-1H-pyrazole-1-yl)-1-(methylsulfonyl)azetidin-3-yl)acetonitrile (200 mg, 56.0% yield) as a yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.26 (br. s., 1H), 8.10 (s, 1H), 7.26-7.28 (m, 2H), 7.24 (d, J=7.28 Hz, 1H), 4.65 (d, J=9.03 Hz, 2H), 4.28 (d, J=9.29 Hz, 2H), 3.45 (s, 2H), 3.04 (s, 3H). MS (ESI) Calcd. for C.sub.16H.sub.16N.sub.6O.sub.3S [M+H].sup.+ 373, Found 373.

Step 8: Preparation of N-(7-(1-(3-(cyanomethyl)-1-(methylsulfonyl)azetidin-3-yl)-1H-pyrazole-4-yl)benzo[d]oxazole-2-yl) cyclopropanecarboxamide(WX30)

(107) To 2-(3-(4-(2-aminobenzo[d]oxazole-7-yl)-1H-pyrazole-1-yl)-1-(methylsulfonyl) azetidin-3-yl)acetonitrile (100 mg, 268.5 umol) and triethylamine (54 mg, 537.1 umol) in acetonitrile (3 mL) was added dropwise cyclopropanecarboxylic acid chloride (30 mg, 287.3 umol) under the protection of nitrogen. The mixture was stirred at 30 C. for reacting for 12 hours. Low yield for the reaction was detected and showed from LCMS. The reaction mixture was directly spun dry, and the residue was separated and purified through preparative TLC (DCM:MeOH=10:1) to give N-(7-(1-(3-(cyanomethyl)-1-(methyl sulfonyl)azetidin-3-yl)-1H-pyrazole-4-yl)benzo[d]oxazole-2-yl) cyclopropanecarboxamide (10 mg, 8.45% yield) as a yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =12.05 (br. s., 1H), 8.77 (s, 1H), 8.34 (s, 1H), 7.60 (d, J=7.53 Hz, 1H), 7.44 (d, J=7.53 Hz, 1H), 7.31-7.38 (m, 1H), 4.52 (d, J=9.29 Hz, 2H), 4.30 (d, J=9.03 Hz, 2H), 3.66 (s, 2H), 3.15 (s, 3H), 2.14 (br. s., 1H), 0.96 (d, J=5.52 Hz, 4H). MS (ESI) Calcd. for C.sub.20H.sub.20N.sub.6O.sub.4S [M+H].sup.+ 441, Found 441.

Example 25

(108) ##STR00057##

Step 1: Preparation of ethyl N-[(3-chloropyrazin 2-yl)thiocarbamoyl]carbamate

(109) To 3-chloropyrazin-2-amine (8.7 g, 67.2 mmol) dissolved in THF (100 mL) was added ethyl isothiocyanate (9.7 g, 73.9 mmol). The resulting mixture was stirred at 27 C. for reacting for 48 hours until LC-MS showed complete reaction. The reaction mixture solvent THF was spun dry under reduced pressure to give a crude product of ethyl N-[(3-chloropyrazin-2-yl)thiocarbamoyl] carbamate which was washed with tert-butyl methyl ether, dried, and directly used in the next step without further purification (purity enough). MS (ESI) Calcd. for C.sub.8H.sub.9ClN.sub.4O.sub.2S [M+H].sup.+ 261, Found 261.

Step 2: Preparation of 8-chloro-[1,2,4]triazolo[1,5-a]pyrazine-2-amine

(110) Hydroxylamine hydrochloride (20.0 g, 287.7 mmol) was suspended in 100 mL of mixture of ethanol and methanol (1:1), and then DIEA (22.3 g, 172.6 mmol) was added. After the resulting mixture was stirred at 27 C. for reacting for 1 hour, ethyl N-[(3-chloropyrazin-2-yl)thiocarbamoyl]carbamate (15.0 g, 57.54 mmol) was added to this system and refluxed (70 C.) slowly for 3 hours. After LC-MS showed that the reaction was completed, the reaction mixture was cooled to room temperature, filtered, washed with water and MTBE, and then dried under vacuum (60 C.) to give 8-chloro-[1,2,4]triazolo[1,5-a]pyrazine-2-amine (6.50 g, 64.62% yield) as a yellow solid. MS (ESI) Calcd. for C.sub.8H.sub.4ClN.sub.5[M+H].sup.+ 170, Found 170.

Step 3: Preparation of N-(8-chloro-[1,2,4]triazolo[1,5-a]pyrazine-2-yl) cyclopropanecarboxamide

(111) To 8-chloro-[1,2,4]triazolo[1,5-a]pyrazine-2-amine (2.0 g, 11.8 mmol) dissolved in anhydrous CH.sub.3CN (30 mL) was added Et.sub.3N (3 g, 29.5 mmol) at 5 C., followed by the addition of cyclopropanecarboxylic acid chloride (3.1 g, 29.5 mmol). After adding, the reaction mixture was warmed to 28 C. and stirred until LC-MS showed that all raw materials are consumed. If needed, Et.sub.3N (7.1 mmol) and cyclopropanecarboxylic acid chloride (7.1 mmol) were further added to ensure complete reaction. The solvent was spun dry under reduced pressure to give the residue. The residue was beat with Et.sub.2O (50 mL), the solid was collected by filtration, washed with H.sub.2O (250 mL), acetone (50 mL) and Et.sub.2O (50 mL), and then dried under vacuum to give N-(8-chloro-[1,2,4]triazolo[1,5-a]pyrazine-2-yl) cyclopropanecarboxamide (1.2 g) as a yellow solid. MS (ESI) Calcd. for C.sub.9H.sub.8ClN.sub.5O [M+H].sup.+ 237, Found 237.

Step 4: Preparation of N-[8-(1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyrazine-2-yl]cyclopropanecarboxamide

(112) To N-(8-chloro-[1,2,4]triazolo[1,5-a]pyrazine-2-yl) cyclopropanecarboxamide (100 mg, 420.8 umol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (122.5 mg, 631.2 umol) and K2CO3 (175 mg, 1.3 mmol), dissolved in the mixture solvent of H.sub.2O (1 mL)/dioxane (4 mL), was added Pd(dppf)Cl.sub.2.CH.sub.2Cl.sub.2 (34 mg, 42.08 umol). After vacuuming and filling with nitrogen, the mixture was stirred at 100 C. for reacting for 3 hours. After LC-MS showed that the reaction was completed, the mixture was cooled to room temperature. The mixture was filtered through diatomite bed and the diatomite was washed with DCM (30 mL). The organic layer was separated and the water layer was extracted with DCM (350 mL). The organic phase was combined, washed with saturated salt water, dried with anhydrous Na.sub.2SO.sub.4. The solvent was spun dry under reduced pressure, and the crude product was purified through thin layer chromatography (DCM/MeoH=10:1) to give N-[8-(1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyrazine-2-yl] cyclopropanecarboxamide (70 mg, 8.1% yield) as a pale yellow solid. MS ((ESI)) Calcd. for C.sub.12H.sub.11N.sub.7O [M+H].sup.+ 270, Found 270.

Step 5: Preparation of N-[8-[1-[3-(cyanomethyl)-1-ethylsulfonylazetidin-3-yl]pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyrazine-2-yl] cyclopropanecarboxamide(WX31)

(113) To N-[8-(1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyrazine-2-yl] cyclopropanecarboxamide (120 mg, 445.7 umol) and 2-(1-ethylsulfonylazetidin-3-ylidene) acetonitrile (125 mg, 668.5 umol) dissolved in DMF (10 mL), was added DBU (136 mg, 891 umol). The resulting mixture was stirred at 40 C. for reacting for 16 hours. After LC-MS showed that the reaction was completed, the solvent was spun dry under reduced pressure, and the residue was dissolved in EtOAc (50 mL). The resulting solution was washed with 1N of HCl (10 mL) and saline water (20 mL), the organic phase was dried with anhydrous Na.sub.2SO.sub.4 and the solvent was spun dry. The residue was purified through preparative thin layer chromatography (PE/EA=1:4) to give a crude compound, which was purified and freeze-dried through preparative HPLC (HCl) to give N-[8-[1-[3-(cyanomethyl)-1-ethylsulfonylazetidin-3-yl]pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]pyrazine-2-yl] cyclopropanecarboxamide (40 mg) as a white solid. .sup.1H-NMR (400 MHz, MeOD-d.sub.4) =8.07 (d, J=8 Hz, 2H), 7.86 (dd, J=7.2, 13.2 Hz, 2H), 7.67 (d, J=8.4 Hz, 2H), 7.44-7.42 (m, 1H), 4.19 (s, 2H), 3.44 (d, J=4.8 Hz, 4H), 3.26 (d, J=4.8 Hz, 4H), 2.95 (m, 1H), 0.87 (m, 2H), 0.74 (m, 2H). MS (ESI) Calcd. for C.sub.19H.sub.21N.sub.9O.sub.3S [M+H].sup.+ 456, Found 456.

Example 26

(114) ##STR00058##

Step 1: Preparation of benzyl 3-oxocyclobutane-carboxylate

(115) 3-Oxocyclobutanecarboxylic acid (3.0 g, 26.3 mmol), benzyl bromide (6.7 g, 39.4 mmol) and potassium carbonate (7.3 g, 52.6 mmol) was dissolved in acetone (30 mL), and then heated to reflux for 10 hours. After TLC showed that the reaction was completed, the reaction mixture was concentrated under reduce pressure to remove the solvent, added water (20 mL), and extracted with ethyl acetate (150 mL2). The combined organic phase was washed with saturated salt water (50 mL), dried with anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue was purified through silica gel column chromatography (petroleum ether/ethyl acetate=10:1) to give benzyl 3-oxocyclobutane-carboxylate (2.5 g, 41.9% yield) as a colorless liquid. MS (ESI) Calcd. for C.sub.12H.sub.12O.sub.3 [M+H].sup.+ 205, Found 205.

Step 2: Preparation of benzyl 3,3-difluorocyclobutane carboxylate

(116) To benzyl 3-oxocyclobutane-carboxylate (1.0 g, 4.9 mmol) in dichloromethane (35 mL), was added dropwise DAST (1.6 g, 9.8 mmol) at 60 C. After adding, the reaction mixture was warmed slowly to 15 C. and stirred for 10 hours. After TLC showed that the reaction was completed, the reaction mixture was cooled to 0 C. Saturated sodium bicarbonate solution (10 mL) for quenching reaction, the water phase was extracted with dichloromethane (30 mL2). The organic phase was combined, washed with saturated salt water (10 mL), dried with anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue was purified through silica gel column chromatography (petroleum ether/ethyl acetate=20:110:1) to give benzyl 3,3-difluorocyclobutane carboxylate (450 mg, 36.54% yield) as a colorless oil. MS (ESI) Calcd. for C.sub.12H.sub.12F.sub.2O.sub.2 [M+H].sup.+ 227, Found 227.

Step 3: Preparation of 3,3-difluorocyclobutane carboxylic acid

(117) To solution of benzyl 3,3-difluorocyclobutane carboxylate (450 mg, 2.0 mmol) dissolved in ethanol (10 mL), was added Pd/C (10%, 40 mg). The reaction mixture was stirred at room temperature for 10 hours under the atmosphere of hydrogen (15 psi). After TLC showed that the reaction was completed, the solid was filtered out, the filtrate was concentrated under vacuum to give 3,3-difluorocyclobutane carboxylic acid (250 mg, 83.1% yield) as a white solid. MS (ESI) Calcd. for C.sub.5H.sub.6F.sub.2O.sub.2[M+H].sup.+ 137, Found 137.

Step 4: Preparation of 3,3-difluorocyclobutane carbonyl chloride

(118) To a solution of 3,3-difluorocyclobutane carboxylic acid (230 mg, 1.69 mmol) and DMF (13 mg, 169.0 umol) dissolved in dichloromethane (5 mL), was added dropwise oxalyl chloride (322 mg, 2.5 mmol) at 0 C. After adding, the reaction mixture was stirred at room temperature for 3 hours. After TLC showed that the reaction was completed, the reaction mixture was concentrated under vacuum to give 3,3-difluorocyclobutane carbonyl chloride (300 mg, a crude product) as a yellow solid. This product was directly used in the next step without further purification. MS (ESI) Calcd. for C.sub.5H.sub.5ClF.sub.2O [M+H].sup.+ 155, Found 155.

Step 5: Preparation of ter-butyl 3-(cyanomethyl)-3-[4-[2-[(3,3-difluorocyclobutane carboxylic acid)amino]-[1,2,4]triazolo[1,5-c]pyrimidin-5-ylpyrazole-1-yl]azetidin-1-carboxylic acid

(119) To a solution of ter-butyl 3-[4-(2-amino-[1,2,4]triazolo[1,5-c]pyrimidin-5-yl) pyrazole-1-yl]-3-(cyanomethyl)-azetidin-1-carboxylic acid (395 mg, 1.0 mmol), DMAP (13 mg, 110 umol) and pyridine (396 mg, 5 mmol), dissolved in dichloromethane (8 mL), was added 3,3-difluorocyclobutane carbonyl chloride (294 mg, 1.9 mmol) at 0 C. The mixture was heated to 40 C. and stirred for 10 hours. The reaction was completed as shown from LCMS. The reaction mixture was poured into water (5 mL) and the water phase was extracted with dichloromethane (15 mL2). The combined organic phase was washed with saturated salt water (10 mL), dried with anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue was purified through preparative TLC (ethyl acetate/petroleum ether=1:0) to give ter-butyl 3-(cyanomethyl)-3-[4-[2-[(3,3-difluorocyclobutanecarboxylic acid)amino]-[1,2,4]triazolo[1,5-c]pyrimidin-5-ylpyrazole-1-yl] azetidin-1-carboxylic acid (80 mg, 12.00% yield) as a white solid. MS (ESI) Calcd. for C.sub.23H.sub.25F.sub.2N.sub.9O.sub.3[M+H].sup.+ 514, Found 514.

Step 6: Preparation of N-[5-[1-[3-(cyanomethyl) azetidin-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]-3,3-difluoro-cyclobutanecarboxamide

(120) To a solution of ter-butyl 3-(cyanomethyl)-3-[4-[2-[(3,3-difluorocyclobutane carboxylic acid)amino]-[1,2,4]triazolo[1,5-c]pyrimidin-5-ylpyrazole-1-yl]azetidin-1-carboxylic acid (80 mg) dissolved in dichloromethane (5 mL) was added dropwise trifluoroacetic acid (765 mg, 6.7 mmol). The reaction mixture was stirred at room temperature for 2 hours. The reaction was completed as shown from LCMS. The reaction mixture was concentrated under vacuum to give N-[5-[1-[3-(cyanomethyl) azetidin-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]-3,3-difluoro-cyclobutanecarboxamide (100 mg, crude product, TFA salt) as a brown solid. This product was directly used in the next step without purification. MS (ESI) Calcd. for C.sub.18H.sub.17F.sub.2N.sub.9O [M+H].sup.+ 414, Found 414.

Step 7: Preparation of N-[5-[1-[3-(cyanomethyl)-1-(trifluoromethylsulfonyl) azetidin-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]-3,3-difluoro-cyclobutanecarboxamide(WX32)

(121) To N-[5-[1-[3-(cyanomethyl)azetidin-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl]-3,3-difluoro-cyclobutanecarboxamide (100 mg, 189.61 umol, TFA salt) in dichloromethane (5 mL) was added trifluoromethanesulfonyl chloride (38 mg, 227.5 mmol), and then triethylamine (96 mg, 948.1 umol) was added. The reaction mixture was stirred at room temperature for 10 hours. The reaction was completed as shown from LCMS. The reaction mixture was concentrated under vacuum, and the residue was purified through preparative HPLC to give N-[5-[1-[3-(cyanomethyl)-1-(trifluoromethylsulfonyl)azetidin-3-yl]pyrazole-4-yl]-[1,2,4]triazolo [1,5-c]pyrimidin-2-yl]-3,3-difluoro-cyclobutanecarboxamide (3 mg, 2.9% yield) as a white solid. MS (ESI) Calcd. for C.sub.19H.sub.16F.sub.5N.sub.9O.sub.3S [M+H].sup.+ 546, Found 546.

Example 27

(122) ##STR00059## ##STR00060##

Step 1: Preparation of tert-butyl tert-butyl 4-[3-(cyanomethyl)-3-[4-[2-(cyclopropylcarbonyl)amino-[1,2,4]triazolo[1,5-a]pyridin-5-yl]pyridylbutane-1-yl]azetidin-1-yl]piperdine-1-carboxylate (1)

(123) To Intermediate 8 (250 mg, 689.9 umol), tert-butyl 4-oxo-piperidine-carboxylate (137 mg, 689.9 umol) and NaBH(OAc).sub.3 (292 mg, 1.4 mmol), dissolved in THF (3 mL), was added DIEA (446 mg, 3.5 mmol). The system is vacuumed and filled with nitrogen. Then the mixture was stirred at 26 C. for reacting for 12 hours. After LC-MS showed that the reaction was completed, the mixture was filtered, and the filtrate was washed with water (3 mL) and then extracted with EA (5 mL3). The organic phase was combined, washed with saturated salt water (3 mL), dried with anhydrous sodium sulfate, filtered, and distilled under reduced pressure to remove the filtrate. The residue was purified through silica gel column chromatography (DCM/MeOH=20/1) to give a yellow solid (143.00 mg, 37.99% yield). MS ESI Calcd. for C.sub.28H.sub.35N.sub.9O.sub.3 [M+H].sup.+ 546, Found 546.

Step 2: Preparation of N-[5-[1-[3-(cyanomethyl)-1-(4-piperdine) azetidin-3-yl]pyrazole-4-yl]-[1,2,4]triazolo[1,5-]pyridin-2-yl]cyclopropanecarboxamide (2)

(124) To tert-butyl tert-butyl 4-[3-(cyanomethyl)-3-[4-[2-(cyclopropylcarbonyl)amino-[1,2,4]triazolo[1,5-a]pyridin-5-yl]pyridylbutane-1-yl]azetidin-1-yl]piperdine-1-carboxylate (143 mg, 262.1 umol) dissolved in dichloromethane (5 mL) was added dropwise trifluoroacetic acid (2 mL). Then the reaction mixture was stirred at 26 C. for reacting for 3 hours. After LC-MS showed that the reaction was completed, the mixture was distilled under reduced pressure to remove the solvent to give a yellow solid (56 mg, crude product). This product was directly used in the next step without purification. MS ESI Calcd. for C.sub.23H.sub.27N.sub.90 [M+H].sup.+ 446, Found 446.

Step 3: Preparation of ethyl ethyl 6-(bromomethyl)-2-(trifluoromethyl)pyrimidin-4-carboxylate (3)

(125) To ethyl 6-methyl-2-(trifluoromethyl)pyrimidin-4-carboxylate (2.0 g, 8.5 mmol) dissolved in acetic acid (12 mL) was added dropwise Br.sub.2 (1.4 g, 8.5 mmol). The reaction mixture was stirred at 80 C. for reacting for 30 min. After LC-MS showed that the reaction was completed, the mixture was distilled under reduced pressure to remove the solvent. The residue was separated through preparation (PE/EA=3/1) to give a yellow oil liquid (610 mg, 12.3% yield). MS ESI Calcd. for C.sub.9H.sub.8BrF.sub.3N.sub.2O.sub.2 [M+H].sup.+ 313, Found 313.

Step 4: Preparation of ethyl ethyl 6-[(dimethylamino)methyl]-2-(trifluoromethyl) pyrimidin-4-carboxylate (4)

(126) To ethyl ethyl 6-(bromomethyl)-2-(trifluoromethyl)pyrimidin-4-carboxylate (610 mg, 2.0 mmol) and N-dimethylmethanamine (318 mg, 3.9 mmol), dissolved in dichloromethane (20 mL), was added dropwise triethylamine (592 mg, 5.9 mmol). The reaction mixture was stirred at 26 C. for 0.5 hour. After LC-MS showed that the reaction was completed, the filtrate was washed with water (20 mL) and then extracted with EA (20 mL3). The organic phase was combined, washed with saturated salt water (20 mL), dried with anhydrous sodium sulfate, filtered, and distilled under reduced pressure to remove the filtrate. The residue was separated and purified through preparation (PE/EA=3/1) to give a yellow liquid (310 mg, 51.6% yield). MS ESI Calcd. for C.sub.11H.sub.14F.sub.3N.sub.3O.sub.2[M+H].sup.+ 278, Found 278.

Step 5: Preparation of 6-[(dimethylamino)methyl]-2-(trifluoromethyl) pyrimidin-4-carboxylic acid (5)

(127) To ethyl ethyl 6-[(dimethylamino)methyl]-2-(trifluoromethyl) pyrimidin-4-carboxylate (310 mg, 1.1 mmol) dissolved in tetrahydrofuran (8 mL) and water (2 mL) was added lithium hydroxide (54 mg, 2.3 mmol). The reaction mixture was stirred at 26 C. for 0.5 hour. After LC-MS showed that the reaction was completed, the mixture was distilled under reduced pressure to remove the solvent to give a yellow oil (314 mg) liquid which was directly used without purification. MS ESI Calcd. for C.sub.9H.sub.10F.sub.3N.sub.3O.sub.2[M+H].sup.+ 250, Found 250.

Step 6: Preparation of N-(5-(1-(3-(cyanomethyl)-1-(1-(6-((dimethylamino)methyl)-2-(trifluoromethyl)pyrimidin-4-carbonyl)piperidine-4-yl)azetidin-3-yl)-1H-pyrazole-4-yl)-[1,2,4]triazolo[1,5-a]cyclopropanecarboxamide(WX33)

(128) To a solution of N-[5-[1-[3-(cyanomethyl)-1-(4-piperidine)azetidin-3-yl]pyrazole-4-yl]-[1,2,4] triazolo[1,5-]pyridine-2-yl] cyclopropanecarboxamide (48 mg, 107.7 umol), the compound 2 (27 mg, 107.7 umol), EDCI (52 mg, 269.4 umol) and HOBt (36 mg, 269.4 umol), dissolved in DMF (3 mL), was added TEA (55 mg, 538.7 umol). The reaction mixture was stirred at 26 C. for reacting for 12 hours. After LC-MS showed that the reaction was completed, the reaction mixture was dissolved in EA (10 mL) and water (10 mL). The organic layer was separated and the water layer was extracted with EA (215 mL) twice. The organic phase was combined, washed with saturated salt water (5 mL), dried with anhydrous sodium sulfate, filtered, distilled under reduced pressure to remove the filtrate, separated through preparation to give a white solid (WX33) (5 mg, 6.9% yield). .sup.1H NMR (400 MHz, METHANOL-d.sub.4) =9.18 (s, 1H), 8.54 (s, 1H), 7.94 (s, 1H), 7.75-7.69 (m, 1H), 7.57 (ddd, J=1.1, 8.1, 18.9 Hz, 2H), 3.87-3.73 (m, 6H), 3.53 (s, 2H), 3.43-3.35 (m, 1H), 3.30-3.21 (m, 1H), 2.74-2.66 (m, 1H), 2.37 (s, 6H), 2.06-1.93 (m, 2H), 1.50 (d, J=10.3 Hz, 1H), 1.38-1.28 (m, 4H), 1.11-1.05 (m, 2H), 1.01-0.95 (m, 2H).sub. MS ESI Calcd. for C.sub.33H.sub.35F.sub.3N.sub.12O.sub.2 [M+H].sup.+ 677, Found 677.

(129) Test for In Vitro Activity of Jak1, Jak2, and Jak3 Kinase

(130) Experimental Materials

(131) Recombinant human protease of JAK1, JAK2 and JAK3 were purchased from Life technology. LANCE Ultra ULight-JAK-1 (Tyr1023) peptide and LANCE Eu-W1024 Anti-phosphotyrosine (PT66) were purchased from PerkinElmer. Multimode ELISA, Envison(PerkinElmer) reader was used.

(132) Experimental Method

(133) The test compound was diluted according to gradient of three times concentration with a final concentration of from 10 uM to 0.17 nM at 11 concentrations totally, each concentration with two complex holes, and the content of DMSO in the detection was 1%.

(134) Enzyme Reaction of JAK1

(135) 2 nM of JAK1 Protein kinase, 50 nM of LANCE Ultra ULight-JAK-1(Tyr1023) peptide, 38 uM of ATP, 50 mM of HEPES (pH 7.5), 10 mM of MgCl.sub.2, 1 mM of EGTA, 2 mM of DTT, 0.01% BRIJ-35. Checker board is White Proxiplate 384-Plus plate (PerkinElmer). The reaction was carried out at room temperature for 90 min and the reaction system was 10 ul.

(136) Enzyme Reaction of JAK2

(137) 0.02 nM of JAK2 Protein kinase, 50 nM of LANCE Ultra ULight-JAK-1(Tyr1023) peptide, 12 uM of ATP, 50 mM of HEPES (pH 7.5), 10 mM of MgCl.sub.2, 1 mM of EGTA, 2 mM of DTT, 0.01% BRIJ-35. Checker board is White Proxiplate 384-Plus plate (PerkinElmer). The reaction was carried out at room temperature for 60 min and the reaction system was 10 ul.

(138) Enzyme Reaction of JAK3

(139) 0.05 nM of JAK3 Protein kinase, 50 nM of LANCE Ultra ULight-JAK-1(Tyr1023) peptide, 4 uM of ATP, 50 mM of HEPES (pH 7.5), 10 mM of MgCl.sub.2, 1 mM of EGTA, 2 mM of DTT, 0.01% BRIJ-35. Checker board is White Proxiplate 384-Plus plate (PerkinElmer). The reaction was carried out at room temperature for 90 min and the reaction system was 10 ul.

(140) Determination for Reaction

(141) 10 ul detection reagent was added to reaction plate, wherein the final concentration of LANCE Eu-W1024 Anti-phosphotyrosine (PT66) was 2 nM, the final concentration of EDTA was 10 nM, incubated at room temperature for 60 min, with Envison reader.

(142) Data Analysis

(143) The reading was converted to inhibition ratio (%) by the following formula: the inhibition ratio (%)=(MinRatio)/(MaxMin)100%. Four parameter curve fitting (Model 205 in XLFIT5, iDBS) measured IC50 data, as shown in Table 1.

(144) TABLE-US-00001 TABLE 1 Compound JAK2 (nM) JAK1/JAK2 (times) JAK3/JAK2 (times) Tofacitinib 4 0.5 0.075 WX00 B F1 F4 WX01 B F2 F3 WX02 A F1 F3 WX03 B F2 F3 WX04 B F1 F3 WX05 B F1 F3 WX06 B F1 F3 WX07 B F2 F4 WX08 A F1 F4 WX09 A F1 F4 WX10 C F1 F1 WX11 C F2 F3 WX12 B F2 F3 WX13 A F3 F4 WX14 A F2 F4 WX15 A F1 F4 WX16 A F1 F3 WX17 B F1 F3 WX18 A F1 F3 WX19 B F1 F3 WX20 A F1 F4 WX21 A F1 F4 WX22 C F1 F3 WX23 C F1 F3 WX24 B F3 F4 WX25 C F3 F2 WX26 C F3 F3 WX27 A F3 F2 WX28 A F3 F4 WX29 A F3 F4 WX30 A F2 F4 WX31 C F2 F3 WX32 C F2 F3 WX33 B F1 F3 A 10 nM; 10 < B 100 nM; 100 < C 1000 nM; 1 F1 5; 5 < F2 10; 10 < F3 25; 25 < F4 100
Conclusion: The selectivity of the compounds of the present invention to JAK2 is superior to that of Tofacitinib.