1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one derivative as Wee1 inhibitor

Abstract

The present invention provides a 1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one compound having an inhibitory effect on Wee1, and includes an application of the compound in treating various types of tumors. ##STR00001##

Claims

1. The compound represented by formula (I) or a pharmaceutically acceptable salt thereof, ##STR00297## wherein, T.sub.1 is N or CH; R.sub.1 is selected from H, halogen, OH, NH.sub.2, or selected from the group consisting of C.sub.1-3 alkyl and C.sub.1-3 heteroalkyl, each of which is optionally substituted by 1, 2 or 3 R; R.sub.2 is H, or selected from the group consisting of C.sub.1-3 alkyl, C.sub.1-3 heteroalkyl and 3-6 membered heterocycloalkyl, each of which is optionally substituted by 1, 2 or 3 R; R.sub.3 is selected from the group consisting of C.sub.3-5 alkenyl and ##STR00298## ring A is selected from the group consisting of phenyl and 5-6 membered heteroaryl, each of which is optionally substituted by 1, 2 or 3 R; R.sub.5 is selected from halogen, OH, NH.sub.2, C(O)NH.sub.2, or selected from the group consisting of C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, C.sub.3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C.sub.3-6 cycloalkyl-O, 3-6 membered heterocycloalkyl-O and ##STR00299## each of which is optionally substituted by 1, 2 or 3 R; R.sub.4 is selected from the group consisting of C.sub.1-6 alkyl, C.sub.3-5 alkenyl, phenyl, and C.sub.1-3 alkyl -phenyl, each of which is optionally substituted by 1, 2 or 3 R; R is selected from F, Cl, Br, I, OH, NH.sub.2, NH(CH.sub.3), N(CH.sub.3).sub.2, Me, Et, CH.sub.2F, CHF.sub.2, CF.sub.3, ##STR00300## the hetero in C.sub.1-3 heteroalkyl, C.sub.1-6 heteroalkyl, 5-6 membered heteroaryl, 3-6 membered heterocycloalkyl is selected from the group consisting of O, S, C(O), C(O)NH, C(O)O, NH and N; In any one of the cases above, the number of the heteroatoms or the heteroatom groups is independently selected from 1, 2 or 3.

2. The compound or the pharmaceutically acceptable salt as defined in claim 1, wherein, R.sub.1 is selected from H, F, Cl, Br, I, OH, NH.sub.2, or selected from the group consisting of C.sub.1-3 alkyl and C.sub.1-3 alkoxyl, each of which is optionally substituted by 1, 2 or 3 R.

3. The compound or the pharmaceutically acceptable salt as defined in claim 2, wherein R.sub.1 is selected from the group consisting of H, F, Cl, Br, I, OH, NH.sub.2, Me and ##STR00301##

4. The compound or the pharmaceutically acceptable salt as defined in claim 1, wherein, R.sub.2 is H, or selected from the group consisting of C.sub.1-3 alkyl, C(O)C.sub.1-3 alkyl, C(O)OC.sub.1-3 alkyl and oxetanyl, each of which is optionally substituted by 1, 2 or 3 R.

5. The compound or the pharmaceutically acceptable salt as defined in claim 4, wherein, R.sub.2 is selected from the group consisting of H, Me, ##STR00302##

6. The compound or the pharmaceutically acceptable salt as defined in claim 1, wherein, ring A is selected from the group consisting of phenyl, pyridinyl, pyrimidyl, thienyl, thiazolyl, and isothiazolyl, each of which is optionally substituted by 1, 2 or 3 R.

7. The compound or the pharmaceutically acceptable salt as defined in claim 6, wherein, ring A is selected from the group consisting of ##STR00303## each of which is optionally substituted by 1, 2 or 3 R.

8. The compound or the pharmaceutically acceptable salt as defined in claim 7, wherein, ring A is selected from the group consisting of ##STR00304##

9. The compound or the pharmaceutically acceptable salt as defined in claim 1, wherein, R.sub.5 is selected from F, Cl, Br, I, OH, NH.sub.2, C(O)NH.sub.2, or selected from the group consisting of C.sub.1-3 alkyl, C.sub.1-3 alkoxyl, C(O)NHC.sub.1-3 alkyl, C.sub.3-6 cycloalkyl, oxetanyl, 2-pyrrolidinonyl, cyclopropyl-O, cyclobutyl-O, oxacyclobutyl-O, oxacyclopentyl-O, azocyclobutyl, 2-oxazolidinonyl, 2-imidazolidinonyl and ##STR00305## each of which is optionally substituted by 1, 2 or 3 R.

10. The compound or the pharmaceutically acceptable salt as defined in claim 9, wherein, R.sub.5 is selected from F, Cl, Br, I, OH, NH.sub.2, C(O)NH.sub.2, or selected from the group consisting of Me, Et, ##STR00306## each of which is optionally substituted by 1, 2 or 3 R.

11. The compound or the pharmaceutically acceptable salt as defined in claim 10, wherein, R.sub.5 is selected from the group consisting of F, Cl, Br, I, OH, NH.sub.2, C(O)NH.sub.2, Me, ##STR00307##

12. The compound or the pharmaceutically acceptable salt as defined in claim 8, wherein, said ##STR00308## is selected from the group consisting of ##STR00309## ##STR00310## ##STR00311##

13. The compound or the pharmaceutically acceptable salt as defined in claim 12, wherein, R.sub.3 is selected from the group consisting of ##STR00312## ##STR00313##

14. The compound or the pharmaceutically acceptable salt as defined in claim 1, wherein, R.sub.4 is selected from the group consisting of C.sub.1-3 alkyl, C.sub.3-5 alkenyl, phenyl, and benzyl, each of which is optionally substituted by 1, 2 or 3 R.

15. The compound or the pharmaceutically acceptable salt as defined in claim 14, wherein, R.sub.4 is selected from the group consisting of Me, ##STR00314## each of which is optionally substituted by 1, 2 or 3 R.

16. The compound or the pharmaceutically acceptable salt as defined in claim 1, wherein, the moiety ##STR00315## is selected from the group consisting of ##STR00316##

17. The compound or the pharmaceutically acceptable salt defined in claim 1 is selected from: wherein, ##STR00317## R.sub.1, R.sub.2, R.sub.5, T.sub.1 and ring A are as defined in claim 1.

18. The compound or the pharmaceutically acceptable salt as defined in claim 17 is selected from the group consisting of ##STR00318## ##STR00319## ##STR00320## wherein, R.sub.1, R.sub.2, R.sub.5 are as defined in claim 17.

19. The compound or the pharmaceutically acceptable salt thereof shown as below is selected from the group consisting of ##STR00321## ##STR00322## ##STR00323## ##STR00324## ##STR00325## ##STR00326## ##STR00327## ##STR00328## ##STR00329## ##STR00330## ##STR00331## ##STR00332## ##STR00333## ##STR00334## ##STR00335## ##STR00336## ##STR00337##

Description

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

(1) The present invention was described in detail herein by the embodiments, but it's not intended to limit the invention. The present invention has been described in detail herein, also disclosed are the specific embodiments thereof, it's apparent for those skilled in the art that modifications and improvements could be done to the specific embodiments of the present invention without departing form the spirit and scope of the present invention.

Intermediate 1

(2) ##STR00132##

(3) Prepared according to the synthetic method in patent WO2007126122.

Intermediate 2

(4) ##STR00133##

(5) Synthetic Route:

(6) ##STR00134## ##STR00135##

Step 1: Synthesis of Compound I2-B

(7) Triethylamine (5.80 g, 57.29 mmol, 7.94 mL) and 4-dimethylaninopyridine (174.97 mg, 1.43 mmol) were slowly added into compound I2-A (3.50 g, 14.32 mmol) and Boc-anhydride (4.69 g, 21.48 mmol, 4.94 mL) in THF solution (40.00 mL) at r.t., the reaction mixture was stirred at 40 C. for 12 hours, then heated to 55 C. and stirred for 18 hours. The crude compound was concentrated up to dryness, purified by silica gel column (PE/EtOAc=6:1, 4:1) to give the compound I2-B. MS m z: 345.5 [M+H].sup.+

Step 2: Synthesis of Compound I2-C

(8) Under argon atmosphere, wet Pd/C (188.05 mg, 159.50 mol, purity 10%) was added into the compound I2-B in THF solution (1.10 g, 3.19 mmol), after replacing the gas in the flask 3 times with hydrogen, the reaction was stirred at 40 C. for 40 hours under 45 Psi. After completion of the reaction, the solution was filtered through diatomite, the filtrate was concentrated to give the crude compound 12-C. MS m/z: 255.1[M+H].sup.+

Step 3: Synthesis of Compound I2-D

(9) Potassium carbonate (626.92 mg, 4.54 mmol) was added into compound I2C (825.00 mg, 3.24 mmol) and p-fluoronitrobenzene (457.16 mg, 3.24 mmol, 343.73 L) in dimethyl sulfoxide solution (10.00 mL) at r.t. The reaction mixture was stirred at 120 C. for 2 hours. The reaction mixture was cooled down to r.t. and slowly added dropwise into water (50 mL) while stirring, solid precipitation appeared, then filtered to give the crude compound I2-D. MS m/z: 376.5 [M+H].sup.+

Step 4: Synthesis of Compound I2-E

(10) Trifluoroacetic acid (7.70 g, 67.53 mmol, 5.00 mL) was added into the compound I2-D (1.25 g, 3.33 mmol) in dichloromethane solution at r.t., the reaction mixture was stirred at 20-25 C. for 40 min. The reaction mixture was concentrated up to dryness, and diluted by 40 mL water, then pH was adjusted to 11-12 with 10% sodium hydroxide solution, the aqueous phase was extracted by 70 mL dichloromethane 3 times, the organic phase was washed by 100 mL saturated brine, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated up to dryness to give the crude compound I2-E. MS m/z: 276.0[M+H].sup.+

Step 5: Synthesis of Compound I2-F

(11) Formaldehyde (1.00 g, 33.41 mmol, 920.52 L), sodium triacetoxyborohydride (1.42 g, 6.68 mmol, 2.00 eq) and acetic acid (1.05 g, 17.49 mmol, 1.00 mL) were added into the compound I2-E (920.00 mg, 3.34 mmol) in methanol (30.00 mL), and was stirred at 20-25 C. for 30 min. The reaction mixture was concentrated, add 5% sodium hydroxide 30 mL and extracted with 50 mL dichloromethane for 3 times, the organic phase was washed by 100 mL saturated brine, dried over anhydrous sodium sulfate, then filtered and concentrated to give the crude compound I2-F. MS m/z: 290.1[M+H].sup.+

Step 6: Synthesis of Compound I2

(12) Under argon atmosphere, wet Pd/C (97.62 mg, 82.80 mol, 10% purity) was added into compound I2-F (800.00 mg, 2.76 mmol) in ethanol (20.00 mL) solution. The gas in the reactor was replaced by hydrogen for 3 times, the mixture was stirred at 20-25 C. and under hydrogen pressure (15 psi) for 32 hours. After the reaction was completed, the reaction mixture was passed through diatomite then filtered and concentrated to give the crude compound 12. MS m/z: 260.1 [M+H].sup.+

Intermediate 3

(13) ##STR00136##

(14) ##STR00137##

Step 1: Synthesis of Compound I3-A

(15) Oxalyl chloride (2.30 g, 24.34 mmol, 1.89 mL) was slowly added dropwise into the compound I2-E (500.00 mg, 1.82 mmol), triethylamine (920.83 mg, 9.10 mmol, 1.26 mL) in DCM (12.00 mL) solution at 0 C., the reaction mixture was stirred for 0.5 hours at 30 C. Water (20 mL) was added into the reaction system, the aqueous phase was extracted by 20 mL DCM for 3 times, the organic phase was washed by 20 mL saturated brine, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated up to dryness to give the crude compound I3-A. MS m/z: 334.1 [M+H].sup.+

Step 2: Synthesis of Compound I3

(16) Ammonium chloride solution (834.33 mg, 15.60 mmol, 545.31 L) and zinc powder (815.95 mg, 12.48 mmol) were added into the compound 13-A (520.00 mg, 1.56 mmol) in water (5.00 mL) and ethanol (50.00 mL) solution, the reaction mixture was stirred at 70 C. for 1 hour. The reaction mixture was filtered and concentrated to give 13. MS m/z: 304.1[M+H].sup.+

Embodiment 1: Compound 1

(17) ##STR00138##

(18) Synthetic Route:

(19) ##STR00139##

Step 1: Synthesis of Compound 1-A

(20) The compound I1 (2.00 g, 9.00 mmol), phenylboronic acid (2.19 g, 18.00 mmol), pyridine (1.42 g, 18.00 mmol) and copper acetate (1.63 g, 9.00 mmol) in N,N-dimethylformamide (40.00 mL) was stirred at 20 C. for 2.0 hours, then stirred at 70 C. for 12 hours. The reaction system was diluted by 100 mL water, then extracted by EtOAc (100 mL2). The organic phases were combined, washed by water (100 mL3), and then washed by saturated brine (100 mL), dried over anhydrous sodium sulfate. The desiccant was filtered off, the solvent was eliminated under reduced pressure to give the crude product. The crude product was purified by column chromatography (PE/EtOAc=5/1) to give the compound 1-A. MS m/z: 298.9[M+H].sup.+

Step 2: Synthesis of Compound 1

(21) m-Chloroperoxybenzoic acid (52.40 mg, 258.08 mol, 85% purity) was added into the compound 1-A (70.00 mg, 234.62 mol) in toluene (5.00 mL) solution. The reaction was stirred at 20 C. for 2 hours. Intermediate 12 (60.86 mg, 234.62 mol) and N,N-diisopropylethylamine (90.97 mg, 703.86 mol) were added into the reaction system. The reaction mixture was stirred at 20 C. for 14 hours. The reaction system was added into sodium thiosulfate (20 mL) while stirring, the aqueous phase was extracted by EtOAc (30 mL3). The organic phases were combined, and washed by saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuum. The crude product was separated by preparative HPLC to give the compound 1. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 8.80 (s, 1H), 7.75-7.31 (m, 7H), 6.85 (d, J=8.8 Hz, 2H), 5.78-5.53 (m, 1H), 5.08 (dd, J=1.2, 10.4 Hz, 1H), 4.92 (dd, J=1.2, 17.2 Hz, 1H), 4.28 (br s, 2H), 3.12-2.92 (m, 4H), 2.35-2.23 (m, 4H), 2.16 (s, 3H), 1.63-1.37 (m, 8H)

(22) MS m/z: 496.1 [M+H].sup.+

Embodiment 2: Compound 2

(23) ##STR00140##

(24) Synthetic Route:

(25) ##STR00141##

Step 1: Synthesis of Compound 2-B

(26) m-Chloroperoxybenzoic acid (82.34 mg, 405.55 mol, 85% purity) was added into the compound 1-A (110.00 mg, 368.682 mmol) in toluene (10.00 mL) solution. The reaction was stirred at 20 C. for 2 hours. Compound 2-A (127.37 mg, 268.68 mol, produced by hydrogenation reduction of I2-D) and N,N-diisopropylethylamine (142.94 mg, 1.11 mmol) were added into the reaction system. The reaction mixture was stirred at 20 C. for 12 hours. The aqueous phase was diluted by sodium hydroxide (30 mL, 0.5 N) and extracted by EtOAc (30 mL2). The organic phases were combined, washed by saturated brine (30 mL) and dried over anhydrous sodium sulfate, filtered and concentrated in vacuum to give the crude product. The crude product was purified by thin phase chromatography (dichloromethane/methanol=10/1) to give the compound 2-B. MS m/z: 596.2[M+H].sup.+

Step 2: Synthesis of Compound 2

(27) Compound 2-B (170.00 mg, 285.36 mol) was dissolved in dichloromethane (3.00 mL), trifluoroacetic acid (1.54 g, 13.51 mmol) was added into the reaction system, then was stirred at 20 C. for 10 min. The crude product was concentrated and separated by preparative HPLC to give the compound 2. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =8.79 (s, 1H), 7.61-7.32 (m, 7H), 6.85-6.82 (d, J=9.2 Hz, 2H), 5.78-5.54 (m, 1H), 5.06 (d, J=9.2 Hz, 1H), 4.95-4.77 (m, 1H), 3.13-2.99 (m, 4H), 2.79-2.65 (m, 4H), 1.61-1.46 (m, 4H), 1.46-1.29 (m, 4H).

(28) MS m/z: 496.1[M+H].sup.+

Embodiment 3: Compound 3

(29) ##STR00142##

(30) Synthetic Route:

(31) ##STR00143##

Step 1: Synthesis of Compound 3-A

(32) Cuprous iodide (599.92 mg, 3.15 mmol), N,N-dimethylethylenediamine (310.99 mg, 3.53 mmol, 379.26 L) and potassium carbonate (600.80 mg, 4.35 mmol) were added separately into compound I1 (700.00 mg, 3.15 mmol) and 2-bromopyridine (497.59 mg, 3.15 mmol, 299.75 L) in dioxane solution. Under nitrogen atmosphere, the reaction mixture was stirred at 95 C. for 48 hours and then added 40 mL ammonia after concentration, then extracted by EtOAc 350 mL (70 mL5), and washed by saturated brine 150 mL, dried over anhydrous sodium sulfate, then filtered to give the crude compound 3-A. MS m/z: 300.0[M+H].sup.+

Step 2: Synthesis of Compound 3

(33) m-Chloroperoxybenzoic acid (88.17 mg, 434.28 mol, 85% purity) was added into compound 3-A (100.00 mg, 334.06 mol) in dichloromethane (10.00 mL) solution, the mixture was stirred at 20-25 C. for 2 hours, then added compound 12 (86.65 mg, 334.06 mol) and N,N-diisopropylethylamine (129.52 mg, 1.00 mmol, 175.03 L), then stirred for 14 hours. The reaction mixture was diluted by dichloromethane 80 mL, then washed separately by saturated sodium bicarbonate and sodium thiosulfate 60 mL (30 mL2), dried over sodium sulfate, then filtered and concentrated to give the crude compound, the crude compound was purified by preparative HPLC (alkaline condition) to give the compound 3. .sup.1H NMR (CHLOROFORM-d, 400 MHz): =8.75 (s, 1H), 8.45 (d, J=5.0 Hz, 1H), 7.74-7.83 (m, 2H), 7.37 (br d, J=9.0 Hz, 2H), 7.11-7.18 (m, 1H), 6.86 (d, J=9.0 Hz, 2H), 5.61 (ddt, J=16.9, 10.4, 6.1 Hz, 1H), 4.95 (d, J=10.0 Hz, 1H), 4.79-4.91 (m, 1H), 4.71 (br d, J=6.0 Hz, 2H), 3.02-3.12 (m, 4H), 2.35 (br s, 4H), 2.25 (s, 3H), 1.51-1.61 (m, 8H)

(34) MS m/z: 511.1[M+H].sup.+

Embodiment 4: Synthesis of Compound 4

(35) ##STR00144##

(36) Synthetic Route:

(37) ##STR00145##

Step 1: Synthesis of Compound 4-A

(38) According to the method for preparing the compound 3-A and the 2-bromopyridine was replaced by 2-bromopyrimidine, compound 4-A crude product was obtained, the crude compound was purified by silica gel column (PE/EtOAc=6/1, 1/1) to deliver the compound 4-A. MS m/z: 300.9 [M+H].sup.+

Step 2: Synthesis of Compound 4

(39) According to the method for preparing the compound 3 and started with the compound 4-A, the crude product of compound 4 was obtained. The crude compound was purified by preparative separation (neutral condition) to give the compound 4. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.84 (d, J=5.0 Hz, 3H), 7.52 (br s, 1H), 7.26-7.26 (m, 1H), 7.23-7.26 (m, 1H), 6.91-6.96 (m, 2H), 5.69 (ddt, J=16.9, 10.3, 6.3 Hz, 1H), 5.03 (dd, J=10.2, 0.9 Hz, 1H), 4.96 (dd, J=17.1, 1.3 Hz, 1H), 4.82 (br d, J=6.3 Hz, 1H), 4.79-4.86 (m, 1H), 3.10-3.17 (m, 4H), 2.42 (br s, 4H), 2.31 (s, 3H), 1.63-1.67 (m, 4H), 1.59 (br t, J=5.5 Hz, 4H)

(40) MS m/z: 512.2[M+H].sup.+

Embodiment 5: Compound 5

(41) ##STR00146##

(42) Synthetic Route:

(43) ##STR00147##

Step 1: Synthesis of Compound 5-A

(44) According to the method for preparing the compound 3-A, and the 2-bromopyridine was replaced by 2-bromoyrazine, the compound 5-A was obtained. MS m/z: 301.0 [M+H].sup.+

Step 2: Synthesis of Compound 5

(45) m-Chloroperoxybenzoic acid (146.01 mg, 719.18 mol, 85% purity) was added into the compound 5-A (135.00 mg, 449.49 mol) in toluene (10.00 mL) solution, the mixture was stirred at 20-25 C. for 2 hours and then added the compound 12 (86.65 mg, 334.06 mol) and N,N-diisopropylethylamine (174.28 mg, 1.35 mmol, 235.51 L), followed by stirring for 13 hours. The reaction mixture was diluted by dichloromethane 80 mL, and washed by saturated sodium bicarbonate 60 mL (30 mL2), dried over anhydrous sodium sulfate, then filtered and concentrated to give the crude compound, the crude compound was purified by preparative HPLC (neutral condition) to give the compound 5. .sup.1H NMR (CHLOROFORM-d, 400 MHz): =9.26 (d, J=1.5 Hz, 1H), 8.77 (s, 1H), 8.37-8.41 (m, 2H), 7.37 (br d, J=8.5 Hz, 2H), 6.90 (d, J=9.0 Hz, 2H), 5.54-5.65 (m, 1H), 4.97 (d, J=9.5 Hz, 1H), 4.87 (d, J=18.1 Hz, 1H), 4.70 (d, J=6.5 Hz, 2H), 3.05-3.12 (m, 4H), 2.41 (br s, 4H), 2.28 (s, 3H), 1.60 (br s, 8H)

(46) MS m/z: 512.1[M+H].sup.+

Embodiment 6: Compound 6

(47) ##STR00148##

(48) ##STR00149##

Step 1: Synthesis of Compound 6-A

(49) Copper acetate (81.72 mg, 449.90 mol) and pyridine (284.70 mg, 3.60 mmol, 290.51 L) were added into I1 (100.00 mg, 449.90 mol) and 5-pyrimidinylboronic acid (111.49 mg, 899.80 mol) in dichloromethane (10.00 mL) solution. Under oxygen atmosphere, the mixture was stirred at 20-25 C. for 32 hours, EtOAc (180 mL), washed by water 240 mL (120 mL2) and saturated brine 120 mL, dried over anhydrous sodium sulfate, then filtered and concentrated to give the crude compound 6-A, which was purified by thin phase chromatography (PE/EtOAc=2/1) to give compound 6-A.

(50) MS m/z: 300.9[M+H].sup.+

Step 2: Synthesis of Compound 6

(51) According to the method for preparing the compound 3 and started with compound 6-A, the crude product of compound 6 was prepared. The crude compound was purified by preparative separation (neutral condition) to give the compound 6. .sup.1H NMR (CDCl.sub.3, 400 MHz): =9.20 (s, 1H), 8.94 (s, 2H), 8.84 (br s, 1H), 7.41 (d, J=9.0 Hz, 2H), 6.96 (d, J=9.0 Hz, 2H), 5.65-5.77 (m, 1H), 5.17 (d, J=10.5 Hz, 1H), 5.05 (d, J=18.1 Hz, 1H), 4.42 (d, J=6.0 Hz, 2H), 3.14-3.20 (m, 4H), 2.49 (br s, 1H), 2.37 (s, 3H), 1.66-1.69 (m, 8H)

(52) MS m/z: 512.1[M+H].sup.+

Embodiment 7: Compound 7

(53) ##STR00150##

(54) Synthetic Route:

(55) ##STR00151##

Step 1: Synthesis of Compound 7-A

(56) According to the method for preparing the compound 6-A, 5-pyrimidinylboronic acid was replaced by 2-thiophenylboronic acid to give the compound 7-A crude product, the crude compound was purified by silica gel column (PE:EtOAc=4:1) to give the compound 7-A. MS m/z: 304.9[M+H].sup.+

Step 2: Synthesis of Compound 7

(57) According to the method for preparing the compound 3 and started with the compound 7-A, the crude product of compound 7 was obtained. The crude compound was purified by preparative HPLC (neutral condition) to give the compound 7. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.81 (s, 1H), 7.34-7.44 (m, 3H), 7.18 (d, J=2.8 Hz, 1H), 7.06 (dd, J=5.5, 3.8 Hz, 1H), 6.88 (br d, J=8.8 Hz, 2H), 5.74 (ddt, J=16.8, 10.5, 5.9 Hz, 1H), 4.97-5.22 (m, 2H), 4.34-4.45 (m, 2H), 3.05-3.17 (m, 4H), 2.41 (br s, 4H), 2.31 (s, 3H), 1.61 (dt, J=19.1, 5.6 Hz, 8H)

(58) MS m/z: 516.0 [M+H].sup.+

Embodiment 8: Compound 8

(59) ##STR00152##

(60) Synthetic Route:

(61) ##STR00153##

Step 1: Synthesis of Compound 7-A

(62) According to the method for preparing the compound 3-A and 2-bromopyridine was replaced by 3-bromothiophene, the crude product of compound 8-A was obtained, the crude compound was purified by silica gel column (PE/EtOAc=10/1, 3/1) to give the compound 8-A. MS m/z: 304.9 [M+H].sup.+

Step 2: Synthesis of Compound 8

(63) According to the method for preparing the compound 3 and started with the compound 8-A, the crude product of compound 8 was obtained. The crude compound was purified by preparative HPLC (neutral condition) to give 8. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.80 (s, 1H), 7.38-7.45 (m, 3H), 7.29-7.33 (m, 1H), 7.20 (br d, J=5.8 Hz, 1H), 6.90 (d, J=9.0 Hz, 2H), 5.71 (dd, J=17.1, 10.3 Hz, 1H), 5.10-5.15 (m, 1H), 4.98-5.10 (m, 1H), 4.41 (br d, J=5.5 Hz, 2H), 3.07-3.17 (m, 4H), 2.38-2.49 (m, 4H), 2.33 (s, 3H), 1.59-1.62 (m, 8H)

(64) MS m/z: 516.0[M+H].sup.+

Embodiment 9: Compound 9

(65) ##STR00154##

(66) Synthetic Route:

(67) ##STR00155##

Step 1: Synthesis of Compound 9-A

(68) According to the method for preparing the compound 3-A and 2-bromopyridine was replaced by 2-bromothiazole, crude product of the compound 9-A was obtained, the crude compound was purified by silica gel column (PE/EtOAc=6/1, 3/1) to give the compound 9-A. MS m/z: 305.9[M+H].sup.+

Step 2: Synthesis of Compound 9

(69) According to the method for preparing the compound 3 and started with the compound 9-A, crude product of compound 9 was obtained, the crude compound was purified by silica gel plate (dichloromethane/methanol=6/1) to give the compound 9. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.75 (s, 1H), 7.49 (br d, J=3.5 Hz, 2H), 7.09 (d, J=3.5 Hz, 1H), 6.91 (br d, J=9.0 Hz, 2H), 5.57-5.70 (m, 1H), 4.98-5.06 (m, 2H), 4.94 (br s, 2H), 3.07-3.13 (m, 4H), 2.38 (br s, 4H), 2.26 (s, 3H), 1.57-1.57 (m, 1H), 1.54-1.54 (m, 1H), 1.54-1.57 (m, 6H)

(70) MS m/z: 517.0[M+H].sup.+

Embodiment 10: Compound 10

(71) ##STR00156##

(72) Synthetic Route:

(73) ##STR00157##

Step 1: Synthesis of Compound 10-A

(74) According to the method for preparing the compound 11-A, and 3-chloro-phenylboronic acid was replaced by 2-chloro-phenylboronic acid, the compound 10-A was obtained. MS m/z: 333.0[M+H].sup.+

Step 2: Synthesis of Compound 10

(75) According to the method for preparing the compound 3, and started with the compound 10-A, crude product of the compound 10 was obtained. The crude compound was separated by preparative HPLC to give the compound 10. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.76 (s, 1H), 7.48-7.52 (m, 1H), 7.28-7.39 (m, 5H), 6.77 (br d, J=9.0 Hz, 2H), 5.59-5.71 (m, 1H), 5.05 (d, J=11.0 Hz, 1H), 4.92 (d, J=18.1 Hz, 1H), 4.11-4.31 (m, 2H), 2.99-3.06 (m, 4H), 2.44 (br s, 4H), 2.29 (s, 3H), 1.56-1.61 (m, 4H), 1.56 (br s, 4H)

(76) MS m/z: 544.0[M+H].sup.+

Embodiment 11: Compound 11

(77) ##STR00158##

(78) Synthetic Route:

(79) ##STR00159##

Step 1: Synthesis of Compound 11-A

(80) At r.t., copper acetate (572.15 mg, 3.15 mmol) and pyridine (498.33 mg, 6.30 mmol, 508.50 L) were added into 11 (700.00 mg, 3.15 mmol) and 3-chloro-phenylboronic acid (738.85 mg, 4.72 mmol) in N,N-dimethylformamide (20.00 mL) solution, and was stirred for 48 hours under oxygen atmosphere, EtOAc (180 mL), washed by water (120 mL2) and saturated brine (120 mL), dried over anhydrous sodium sulfate, then filtered and concentrated to give the crude compound 11-A. MS m/z: 332.9 [M+H].sup.+

Step 2: Synthesis of Compound 11

(81) According to the method for preparing the compound 3, and started with the compound 11-A, crude product of the compound 11 was obtained. The crude compound was purified by preparative HPLC to give the compound 11. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.72 (s, 1H), 7.41-7.52 (m, 4H), 7.29-7.36 (m, 2H), 6.92-6.94 (br d, J=9.0 Hz, 2H), 5.56-5.72 (m, 1H), 5.10 (d, J=11.0 Hz, 1H), 5.00 (d, J=18.1 Hz, 1H), 4.39-4.40 (m, 2H), 3.12-3.15 (m, 4H), 2.48 (br s, 4H), 2.35 (s, 3H), 1.50-1.61 (m, 8H)

(82) MS m/z: 544.0[M+H].sup.+

Embodiment 12: Compound 12

(83) ##STR00160##

(84) Synthetic Route:

(85) ##STR00161##

Step 1: Synthesis of Compound 12-A

(86) According to the method for preparing the compound 11-A, and 3-chloro-phenylboronic acid was replaced by 4-chloro-phenylboronic acid, crude product of the compound 10-A was obtained, the crude compound was purified by silica gel (PE:EtOAc=10:1-5:1) to give the 12-A. MS m/z: 332.9[M+H].sup.+

Step 2: Synthesis of Compound 12

(87) According to the method for preparing the compound 3, and started with the compound 12-A, crude product of the compound 12 was obtained, the crude compound was purified by preparative HPLC (neutral condition) to give the compound 12.

(88) .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.82 (s, 1H), 7.47-7.52 (m, 2H), 7.37-7.46 (m, 4H), 6.92 (d, J=9.0 Hz, 2H), 5.65-5.75 (m, 1H), 5.13 (d, J=10.0 Hz, 1H), 5.01 (dd, J=17.1, 1.0 Hz, 1H), 4.38 (br d, J=5.5 Hz, 2H), 3.08-3.23 (m, 4H), 2.44 (br s, 4H), 2.33 (s, 3H), 1.62 (br t, J=5.8 Hz, 8H)

(89) MS m/z: 544.1[M+H].sup.+

Embodiment 13: Compound 13

(90) ##STR00162##

(91) Synthetic Route:

(92) ##STR00163##

Step 1: Synthesis of Compound I3-A

(93) According to the method for preparing the compound 3-A and 2-bromopyridine was replaced by 6-bromo-2-ethylpicolinate, crude product of the compound 13-A was obtained, the crude compound was purified by silica gel column (PE/EtOAc=5/1) to give the compound 13-A. MS m/z: 372.0 [M+H].sup.+

Step 2: Synthesis of Compound 13-B

(94) According to the method for preparing the compound 5, and started with the compound 13-A, crude product of the compound 13-B (yellow solid, 100 mg) was obtained. MS m/z: 583 [M+1].sup.+

Step 3: Synthesis of Compound 13

(95) According to the method for preparing the compound 14, methylamine solution was replaced by ammonia, crude product of the compound 13 was obtained, the crude compound was purified by preparative HPLC (neutral condition) to give the compound 13. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.83 (s, 1H), 8.11-8.16 (m, 1H), 8.00 (d, J=3.8 Hz, 2H), 7.79 (br d, J=5.0 Hz, 1H), 7.41-7.42 (m, 1H), 7.41-7.42 (m, 1H), 6.92 (d, J=9.0 Hz, 2H), 5.72 (ddt, J=16.9, 10.4, 6.1 Hz, 1H), 5.08 (dd, J=10.3, 1.0 Hz, 1H), 4.97 (dd, J=17.1, 1.3 Hz, 1H), 4.65 (d, J=6.3 Hz, 2H), 3.11-3.20 (m, 4H), 3.03-3.06 (m, 3H), 2.41 (br s, 4H), 2.30 (s, 3H), 1.51-1.71 (m, 8H)

(96) MS m/z: 568.1 [M+H].sup.+

Embodiment 14: Compound 14

(97) ##STR00164##

(98) Synthetic Route:

(99) ##STR00165##

Step 1: Synthesis of Compound 14

(100) Methylamine solution (5.00 mL, 27.5% purity) was added into the compound 13-B (50.00 mg, 85.81 mol) in methanol (3 mL) solution, the mixture was stirred at 20-25 C. for 4.5 hours and concentrated under reduced pressure to give the crude compound, the product was purified by preparative HPLC (neutral condition) to give the compound 14. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.83 (s, 1H), 8.11-8.16 (m, 1H), 8.00 (d, J=3.8 Hz, 2H), 7.79 (br d, J=5.0 Hz, 1H), 7.41-7.42 (m, 1H), 7.41-7.42 (m, 1H), 6.92 (d, J=9.0 Hz, 2H), 5.72 (ddt, J=16.9, 10.4, 6.1 Hz, 1H), 5.08 (dd, J=10.3, 1.0 Hz, 1H), 4.97 (dd, J=17.1, 1.3 Hz, 1H), 4.65 (d, J=6.3 Hz, 2H), 3.11-3.20 (m, 4H), 3.03-3.06 (m, 3H), 2.41 (br s, 4H), 2.30 (s, 3H), 1.51-1.71 (m, 8H)

(101) MS m/z: 568.1[M+1].sup.+

Embodiment 15: Compound 15

(102) ##STR00166##

(103) Synthetic Route:

(104) ##STR00167##

Step 1: Synthesis of Compound 15

(105) According to the method for preparing the compound 14, and the methylamine solution was replaced by dimethylamine solution, crude product of the compound 15 was obtained, and the crude compound was purified by preparative HPLC (neutral condition) to give the compound 15. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.82 (s, 1H), 7.90-8.00 (m, 2H), 7.53-7.56 (m, 1H), 7.43 (br d, J=8.8 Hz, 2H), 6.93 (d, J=9.0 Hz, 2H), 5.67 (dd, J=17.1, 10.3 Hz, 2H), 5.03 (d, J=10.3 Hz, 1H), 4.95 (dd, J=17.1, 1.3 Hz, 1H), 4.76 (br d, J=5.8 Hz, 2H), 3.14-3.17 (m, 6H), 2.44 (br s, 4H), 2.32 (s, 3H), 1.58-1.65 (m, 8H)

(106) MS m/z: 582.1[M+H].sup.+

Embodiment 16: Compound 16

(107) ##STR00168##

(108) Synthetic Route:

(109) ##STR00169##

Step 1: Synthesis of Compound 16-A

(110) According to the method for preparing the compound 6-A, and the 5-pyrimidinylboronic acid was replaced by 3-methoxycarbonylphenylboronic acid, compound 16-A was obtained. MS m/z: 357.2 [M+H].sup.+

Step 2: Synthesis of Compound 16-B

(111) The compound 16-A (150.00 mg, 420.88 mol) was added into ammonia 5 mL, and was stirred at 20-25 C. for 14 hours, then concentrated to give the crude compound, the product was purified by thin phase chromatography (PE/EtOAc=8/5) to give the compound 16-B. MS m/z: 342.0[M+H].sup.+

Step 3: Synthesis of Compound 16

(112) According to the method for preparing the compound 3, and started with the compound 16-B, crude product of the compound 16 was obtained, the crude compound was purified by HPLC (neutral condition) to give the compound 16. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.81 (s, 1H), 7.95 (s, 1H), 7.81 (d, J=7.0 Hz, 1H), 7.50-7.64 (m, 3H), 7.40 (br d, J=8.8 Hz, 2H), 6.90 (d, J=8.8 Hz, 2H), 5.65-5.65 (m, 1H), 5.65-5.71 (m, 1H), 5.11 (d, J=11.3 Hz, 1H), 4.99 (d, J=17.3 Hz, 1H), 4.41 (br d, J=5.5 Hz, 2H), 3.10-3.15 (m, 4H), 2.44 (br s, 4H), 2.33 (s, 3H), 1.65 (br d, J=6.0 Hz, 8H)

(113) MS m/z: 553.1[M+H].sup.+

Embodiment 17: Compound 17

(114) ##STR00170##

(115) Synthetic Route:

(116) ##STR00171##

Step 1: Synthesis of Compound 17-A

(117) According to the method for preparing the compound 3-A and the 2-bromopyridine was replaced by methyl 3-bromobenzoate, the compound 17-A was obtained. MS m/z: 357.2[M+H].sup.+

Step 2: Synthesis of Compound 17

(118) Dimethylamine solution (4.45 g, 32.57 mmol, 5.00 mL, 33% purity) was added into the compound 17-A (165.00 mg, 290.66 mol) in methanol (5.00 mL) solution. After stirring at 20-25 C. for 32 hours, the reaction mixture was concentrated, diluted by water 25 mL, extracted by dichloromethane 90 mL (30 mL3), the organic phase was washed by saturated brine 40 mL, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude compound, the crude compound was purified by preparative HPLC (alkaline condition) to give the compound 17. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.73 (s, 1H), 7.44-7.49 (m, 2H), 7.38-7.42 (m, 1H), 7.34 (br dd, J=8.3, 4.3 Hz, 2H), 7.31-7.37 (m, 1H), 6.82 (d, J=9.0 Hz, 2H), 5.56-5.68 (m, 1H), 5.03 (d, J=10.0 Hz, 1H), 4.93 (d, J=17.1 Hz, 1H), 4.67 (br s, 4H), 4.32 (br d, J=6.0 Hz, 2H), 3.06 (br t, J=5.5 Hz, 6H), 2.46 (br s, 4H), 2.31 (s, 3H), 1.59 (br t, J=5.3 Hz, 8H)

(119) MS m/z: 581.1[M+H].sup.+

Embodiment 18: Compound 18

(120) ##STR00172##

(121) ##STR00173##

Step 1: Synthesis of Compound 18-A

(122) Ammonium formate (8.45 g, 134.10 mmol), (1,1-bis(diphenylphosphino)ferrocene) palladium dichloride*dichloromethane (1.64 g, 2.01 mmol) were added into the compound 1-A (4.00 g, 13.41 mmol) in THF (80.00 mL) solution. Under nitrogen atmosphere, the reaction mixture was stirred at 80 C. for 12 hours. The reaction was cooled down to 20 C., then filtered, the residue was washed by THF (30 mL) once to give crude product. Water (100 mL) was added into the crude product while stirring, the suspension was filtered and the residue was washed by water (50 mL) to give a pale yellow solid 18-A. MS m/z: 258.9 [M+H].sup.+

Step 2: Synthesis of Compound 18-B

(123) Potassium hydroxide (81.46 mg, 1.45 mmol) and methyl iodide (1.00 g, 7.05 mmol, 438.60 L) were added into the compound 18-A (250.00 mg, 967.87 mol, 1) in ethanol (20.00 mL) solution. The reaction mixture was stirred at 80 C. for 23 hours. The reaction mixture directly evaporated to give the crude product. The crude compound was purified by silica gel column (PE/EtOAc=3/1) to give 18-B. .sup.1H NMR (400 MHz, CDCl.sub.3) 2.50 (s, 3H) 3.36 (s, 3H) 7.26 (s, 1H) 7.39-7.45 (m, 3H) 7.50-7.57 (m, 2H) 8.91 (s, 1H)

(124) MS m/z: 272.9 [M+H].sup.+

Step 3: Synthesis of Compound 18

(125) m-Chloroperoxybenzoic acid (67.10 mg, 330.48 mol, 85% purity) was added into the compound 18-B (60.00 mg, 220.32 mol) in toluene (8.00 mL) solution. The reaction was stirred at 25 C. for 2 hours. Intermediate 12 (74.29 mg, 286.42 mol) and N,N-diisopropylethylamine (85.42 mg, 660.96 mol) were added into the reaction system. The reaction mixture was stirred at 25 C. for 12 hours. Sodium thiosulfate solution (30 mL) was added into the reaction system to quench the extra m-chloroperoxybenzoic acid, then extracted with EtOAc (30 mL3). The organic phases were combined and washed by saturated brine (50 mL), dried over anhydrous sodium sulfate. The desiccant was filtered off, the solvent was eliminated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (neutral) to give the compound 18. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.62 (dt, J=18.76, 5.55 Hz, 8H) 2.32 (s, 3H) 2.44 (br s, 4H) 3.09-3.14 (m, 4H) 3.31 (s, 3H) 6.89 (d, J=9.03 Hz, 2H) 7.38-7.46 (m, 5H) 7.50-7.55 (m, 2H) 8.80 (s, 1H)

(126) MS m/z: 484.1 [M+H].sup.+

Embodiment 19: Compound 19

(127) ##STR00174##

(128) Synthetic Route:

(129) ##STR00175##

Step 1: Synthesis of Compound 19-A

(130) According to the method for preparing the compound 18-B, and methyl iodide was replaced by bromoethane, crude product of the compound 19-A was obtained. The crude compound 19-A was purified by preparative TLC (PE/EtOAc=3/1) to give 19-A. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.09 (t, J=7.15 Hz, 3H) 2.50 (s, 3H) 3.91 (q, J=7.03 Hz, 2H) 7.39-7.45 (m, 3H) 7.50-7.56 (m, 2H) 8.90 (s, 1H)

(131) MS m/z: 286.9 [M+H].sup.+

Step 2: Synthesis of Compound 19

(132) According to the method for preparing the compound 18, and started with the compound 19-A, compound 19 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.00 (t, J=6.78 Hz, 3H) 1.50-1.60 (m, 8H) 2.24 (s, 3H) 2.35 (br s, 4H) 3.03-3.07 (m, 4H) 3.80 (q, J=6.69 Hz, 2H) 6.82 (d, J=9.03 Hz, 2H) 7.31-7.41 (m, 5H) 7.43-7.48 (m, 2H) 8.72 (s, 1H)

(133) MS m/z: 498.1 [M+H].sup.+

Embodiment 20: Compound 20

(134) ##STR00176##

(135) Synthetic Route:

(136) ##STR00177##

Step 1: Synthesis of Compound 20-A

(137) According to the method for preparing the compound 18-B, and methyl iodide was replaced by bromomethylcyclopropane, compound 20-A was obtained. MS m/z: 313.0 [M+H].sup.+

Step 2: Synthesis of Compound 20

(138) According to the method for preparing the compound 18, and started with the compound 20-A, compound 20 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 0.11-0.13 (m, 2H) 0.36-0.41 (m, 2H) 0.86-0.90 (m, 1H) 1.69 (br s, 8H) 2.43 (s, 3H) 2.60 (br s, 4H) 3.10-3.14 (m, 4H) 3.70 (br d, J=6.78 Hz, 2H) 4.41 (br s, 1H) 6.89 (br d, J=9.28 Hz, 2H) 7.38 (br d, J=7.04 Hz, 1H) 7.43-7.48 (m, 1H) 7.46 (br d, J=7.54 Hz, 3H) 7.51 (br d, J=7.54 Hz, 2H) 8.81 (s, 1H)

(139) MS m/z: 524.1 [M+H].sup.+

Embodiment 21: Compound 21

(140) ##STR00178##

(141) Synthetic Route:

(142) ##STR00179##

Step 1: Synthesis of Compound 21-A

(143) According to the method for preparing the compound 18-B, and methyl iodide was replaced by 1-bromo-2-methoxyethane, compound 21-A was obtained. MS m/z: 317.0 [M+H].sup.+

Step 2: Synthesis of Compound 21

(144) According to the method for preparing the compound 18, and started with compound 21-A, compound 21 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.59-1.65 (m, 8H) 2.31 (s, 3H) 2.43 (br s, 4H) 3.10-3.14 (m, 4H) 3.23 (s, 3H) 3.46 (t, J=5.66 Hz, 2H) 4.01 (br t, J=5.40 Hz, 2H) 6.89 (d, J=8.78 Hz, 2H) 7.41-7.53 (m, 7H) 8.80 (s, 1H)

(145) MS m/z: 528.1 [M+H].sup.+

Embodiment 22: Compound 22

(146) ##STR00180##

(147) Synthetic Route:

(148) ##STR00181## ##STR00182##

Step 1: Synthesis of Compound 22-B

(149) Compound benzyl triethyl ammonium chloride (117.25 mg, 514.76 mol), potassium carbonate (1.58 g, 11.44 mmol) and compound 1-bromo-2,2-difluoroethane (1.20 g, 8.29 mmol) were added into the compound 22-A (1.50 g, 5.72 mmol, obtained when preparing the Intermediate I1) in acetonitrile (15.00 mL) solution. The reaction mixture was stirred at 50 C. for 12 hours. The reaction mixture was evaporated to give the crude product, water (20 mL) was added into the crude product and extracted by EtOAc (30 mL3). The organic phases were combined and washed by saturated brine (50 mL), dried over anhydrous sodium sulfate. The desiccant was filtered off, the solvent was eliminated under reduced pressure to give the crude product. The crude compound was purified by silica gel column (PE/EtOAc=5/1) to give the compound 22-B. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.36 (s, 6H) 1.56 (s, 3H) 3.95-4.08 (m, 2H) 5.90-6.24 (m, 1H) 7.79-7.87 (m, 2H) 7.94 (td, J=6.08, 3.14 Hz, 2H)

Step 2: Synthesis of Compound 22-C

(150) Methyl hydrazine (316.73 mg, 2.75 mmol, 359.92 L, 40% purity) was added into the compound 22-B (360.00 mg, 1.10 mmol) in THF (8.00 mL) solution at 0-5 C. The reaction mixture was stirred at 40 C. for 12 hours. The reaction mixture was cooled down to 25 C., then filtered, the filtrate was evaporated to give the crude product, hexane/EtOAc=3/1 (24 mL) was added into the crude product and stirred for 5 min, then filtered, the filtrate was concentrated and evaporated to dry to give the compound 22-C. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.48 (s, 9H) 3.68-3.78 (m, 2H) 5.75-6.11 (m, 1H)

Step 3: Synthesis of Compound 22-D

(151) Compound 4-chloro-2-(methylthio)pyrimidin-5-ethyl formate (265.00 mg, 1.14 mmol) and N,N-diisopropylethylamine (442.00 mg, 3.42 mmol, 597.30 L) were added into the compound 22-C (223.66 mg, 1.14 mmol) in THF (25.00 mL) solution. The reaction mixture was stirred at 80 C. for 12 hours. The crude product was diluted by EtOAc (30 mL) while stirring for 5 min, then filtered, the filtrate was evaporated to dry to give the compound 22-D. MS m/z: 393.1 [M+H].sup.+

Step 4: Synthesis of Compound 22-E

(152) Trifluoroacetic acid (10.00 mL) was slowly added dropwise into the compound 22-D (205.00 mg, 522.40 mol) at 0-5 C., the reaction mixture was stirred at 25 C. for 4 hours. The reaction mixture was evaporated to dry to give the compound 22-E.

(153) MS m/z: 292.9 [M+H].sup.+

Step 5: Synthesis of Compound 22-F

(154) At 0-5 C., sodium hydroxide solution (4 M, 2.00 mL) was added into the compound 22-E (700.00 mg, 2.39 mmol) in ethanol (8.00 mL) solution. The reaction mixture was stirred at 25 C. for 30 min. The reaction mixture was adjusted to pH=1 by diluted hydrochloric acid, then extracted by dichloromethane (50 mL6), the organic phases were combined and washed by saturated brine (50 mL), dried over anhydrous sodium sulfate. The desiccant was filtered off, the solvent was eliminated under reduced pressure to give the compound 22-F. MS m/z: 247.0 [M+H].sup.+

Step 6: Synthesis of Compound 22-G

(155) According to the method for preparing the compound 3-A and 2-bromopyridine was replaced by bromobenzene, crude product of the compound 22-G was obtained. The crude product was purified by column chromatography (PE/EtOAc=5/1, 3/1) to give the compound 22-G. MS m/z: 323.0 [M+H].sup.+

Step 7: Synthesis of Compound 22

(156) m-Chloroperoxybenzoic acid (54.67 mg, 269.29 mol, 85% purity) was added into the compound 22-G (62.00 mg, 192.35 mol) in toluene (5.00 mL) solution. The reaction was stirred at 30 C. for 1 hour. Intermediate 12 (74.84 mg, 288.52 mol) and N,N-diisopropylethylamine (74.58 mg, 577.05 mol) were added into the reaction system. The reaction mixture was stirred at 30 C. for 12 hours. Water (20 mL) was added into the reaction system, then extracted by EtOAc (20 mL3). The organic phases were combined, the organic phase was quenched by saturated sodium bicarbonate (30 mL) for the extra m-chloroperoxybenzoic acid and then washed by saturated brine (30 mL), dried over anhydrous sodium sulfate. The desiccant was filtered off, the solvent was eliminated under reduced pressure to give the crude product, which was purified by preparative HPLC (neutral) to give to give the compound 22. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.64 (dt, J=19.02, 5.55 Hz, 8H) 2.34 (s, 3H) 2.44 (br s, 4H) 3.09-3.20 (m, 4H) 4.06-4.19 (m, 2H) 5.83-6.16 (m, 1H) 6.90 (br d, J=8.78 Hz, 2H) 7.45 (br d, J=7.78 Hz, 4H) 7.50-7.63 (m, 3H) 8.84 (s, 1H)

(157) MS m/z: 534.0 [M+H].sup.+

Embodiment 23: Compound 23

(158) ##STR00183##

(159) Synthetic Route:

(160) ##STR00184##

Step 1: Synthesis of Compound 23-A

(161) According to the method for preparing the compound 18-B and methyl iodide was replaced by benzyl bromide, the compound 23-A was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 2.45 (s, 3H) 5.01 (s, 2H) 6.98 (dd, J=7.78, 1.76 Hz, 2H) 7.17-7.24 (m, 3H) 7.27-7.31 (m, 2H) 7.39-7.44 (m, 1H) 7.47-7.52 (m, 2H) 8.91 (s, 1H)

(162) MS m/z: 348.9 [M+H].sup.+

Step 2: Synthesis of Compound 23

(163) According to the method for preparing the compound 18, and started with the compound 23-A, the compound 23 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.61 (dt, J=19.64, 5.36 Hz, 8H) 2.31 (s, 3H) 2.41 (br s, 4H) 3.06-3.14 (m, 4H) 4.94-4.98 (m, 1H) 4.96 (s, 2H) 6.85 (br d, J=8.54 Hz, 2H) 6.98-7.06 (m, 2H) 7.16-7.24 (m, 3H) 7.32-7.41 (m, 5H) 7.47-7.52 (m, 2H) 8.80 (s, 1H)

(164) MS m/z: 560.1 [M+H].sup.+

Embodiment 24: Compound 24

(165) ##STR00185##

(166) Synthetic Route:

(167) ##STR00186##

Step 1: Synthesis of Compound 24-A

(168) According to the method for preparing the compound 3-A and 2-bromopyridine was replaced by 2-(6-bromopyridin-2-yl)isopropanol, the compound 24-A was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.59 (s, 6H) 2.58 (s, 3H) 3.71 (s, 1H) 4.81 (br d, J=6.02 Hz, 2H) 4.90-4.96 (m, 1H) 5.06 (br d, J=10.29 Hz, 1H) 5.69 (ddt, J=16.81, 10.42, 6.21, 6.21 Hz, 1H) 7.40 (d, J=7.78 Hz, 1H) 7.76 (d, J=7.78 Hz, 1H) 7.88-7.94 (m, 1H) 8.94 (s, 1H)

(169) MS m/z: 358.0 [M+H].sup.+

Step 2: Synthesis of Compound 24-B

(170) Pd(OH).sub.2/C (100.00 mg, 131.66 mol, 20% purity) was added into the compound 24-A (100.00 mg, 279.78 mol) in ethanol (8.00 mL) solution. The reaction mixture was stirred under hydrogen (15 Psi) at 25 C. for 12 hours. The reaction mixture was filtered, the filtrate was concentrated under reduced pressure to remove the solvent so as to give the compound 24-B.

(171) MS m/z: 360.2 [M+H].sup.+

Step 3: Synthesis of Compound 24

(172) According to the method for preparing the compound 18, and started with compound 24-B, the compound 24 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 0.66 (t, J=7.28 Hz, 3H) 1.34-1.40 (m, 2H) 1.45 (s, 6H) 1.48-1.54 (m, 8H) 2.19 (s, 3H) 2.31 (br s, 4H) 3.00-3.04 (m, 4H) 3.99 (t, J=7.04 Hz, 2H) 6.81 (d, J=9.04 Hz, 2H) 7.24 (d, J=7.54 Hz, 1H) 7.33 (br d, J=9.04 Hz, 2H) 7.63 (d, J=8.04 Hz, 1H) 7.73-7.79 (m, 1H) 8.70 (s, 1H)

(173) MS m/z: 571.1 [M+H].sup.+

Embodiment 25: Compound 25

(174) ##STR00187##

(175) Synthetic Route:

(176) ##STR00188##

Step 1: Synthesis of Compound 25-A

(177) According to the method for preparing the compound 27-A in embodiment 27 except for the corresponding starting material 2-bromo-6-chloropyridine, compound 25-A was obtained. MS m/z: 334.0 [M+H].sup.+

Step 2: Synthesis of Compound 25

(178) According to the method for preparing the compound 22, and started with the compound 25-A, the compound 25 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.61-1.71 (m, 8H) 2.35 (s, 3H) 2.45-2.56 (m, 4H) 3.11-3.19 (m, 4H) 4.80 (br d, J=6.28 Hz, 2H) 4.97-5.06 (m, 2H) 5.69 (ddt, J=16.86, 10.32, 6.24, 6.24 Hz, 1H) 6.88-6.95 (m, 2H) 7.20-7.25 (m, 1H) 7.42-7.48 (m, 2H) 7.75-7.80 (m, 1H) 7.82-7.86 (m, 1H) 8.82 (s, 1H)

(179) MS m/z: 545.0 [M+H].sup.+

Embodiment 26: Compound 26

(180) ##STR00189##

(181) ##STR00190##

Step 1: Synthesis of Compound 26-A

(182) According to the method for preparing the compound 27-A, 2-bromo-6-fluoropyridine was replaced by 2-bromo-5-chloropyridine, crude product of the compound 26-A was obtained, the crude product was purified by column chromatography (PE/EtOAc=5/1, 3/1) to give the compound 26-A. MS m/z: 333.9 [M+H].sup.+

Step 2: Synthesis of Compound 26

(183) According to the method for preparing the compound 22, and started with the compound 26-A, the compound 26 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.51-1.62 (m, 8H) 2.23 (s, 3H) 2.34 (br s, 4H) 3.07-3.11 (m, 4H) 4.69 (br d, J=6.02 Hz, 2H) 4.87 (d, J=17.08 Hz, 1H) 4.96 (d, J=9.54 Hz, 1H) 5.59 (ddt, J=16.88, 10.35, 6.46, 6.46 Hz, 1H) 6.87 (d, J=9.04 Hz, 2H) 7.35 (br d, J=9.04 Hz, 2H) 7.69-7.75 (m, 1H) 7.76-7.90 (m, 1H) 8.38 (d, J=2.52 Hz, 1H) 8.75 (s, 1H)

(184) MS m/z: 545.0 [M+H].sup.+

Embodiment 27: Compound 27

(185) ##STR00191##

(186) Synthetic Route:

(187) ##STR00192##

Step 1: Synthesis of Compound 27-A

(188) Potassium carbonate (213.62 mg, 1.55 mmol), CuI (213.30 mg, 1.12 mmol, 1.00 eq) and N,N-dimethylethylenediamine (110.58 mg, 1.25 mmol, 134.85 L) were added into compound I1 (250.00 mg, 1.12 mmol) and 2-bromo-6-fluoropyridine (203.02 mg, 1.15 mmol) in dioxane (8.00 mL) solution. The reaction mixture was stirred under nitrogen atmosphere at 95 C. for 1 hour. The reaction mixture was cooled down and then ammonia (30 mL) was added, extracted by EtOAc (50 mL3), the organic phases were combined, the organic phase was washed by saturated brine (50 mL) once, dried over anhydrous sodium sulfate, then filtered, the filtrate was evaporated to give the compound 27-A. MS m/z: 318.0 [M+H].sup.+

Step 2: Synthesis of Compound 27

(189) According to the method for preparing the compound 22, and started with the compound 27-A, the compound 27 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.62 (br t, J=5.52 Hz, 8H) 2.33 (s, 3H) 2.43 (br s, 4H) 3.15-3.20 (m, 4H) 4.82 (d, J=6.54 Hz, 2H) 4.98-5.08 (m, 2H) 5.71 (ddt, J=16.94, 10.42, 6.16, 6.16 Hz, 1H) 6.85 (dd, J=8.04, 2.52 Hz, 1H) 6.96 (d, J=9.04 Hz, 2H) 7.47 (br d, J=8.54 Hz, 2H) 7.83 (d, J=8.04 Hz, 1H) 7.94 (q, J=7.70 Hz, 1H) 8.84 (s, 1H)

(190) MS m/z: 529.1 [M+H].sup.+

Embodiment 28: Compound 28

(191) ##STR00193##

(192) Synthetic Route:

(193) ##STR00194##

Step 1: Synthesis of Compound 28-A

(194) According to the method for preparing the compound 27-A, the 2-bromo-6-fluoropyridine was replaced by 2-bromo-5-fluoropyridine, crude product of the compound 28-A was obtained, the crude product was purified by column chromatography (PE/EtOAc=5/1, 3/1) to give a yellow compound 28-A. MS m/z: 318.0 [M+H].sup.+

Step 2: Synthesis of Compound 28

(195) According to the method for preparing the compound 22, and started with the compound 28-A, the compound 28 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.60-1.70 (m, 8H) 2.32 (s, 3H) 2.42 (br s, 4H) 3.13-3.21 (m, 4H) 4.75 (br d, J=6.54 Hz, 2H) 4.95 (dd, J=17.08, 1.00 Hz, 1H) 5.04 (d, J=10.04 Hz, 1H) 5.69 (ddt, J=16.88, 10.36, 6.46, 6.46 Hz, 1H) 6.94 (d, J=8.54 Hz, 2H) 7.43 (d, J=9.04 Hz, 2H) 7.60 (td, J=8.16, 2.76 Hz, 1H) 7.89 (br dd, J=8.78, 3.76 Hz, 1H) 8.38 (d, J=2.52 Hz, 1H) 8.84 (s, 1H)

(196) MS m/z: 529.0 [M+H].sup.+

Embodiment 29: Compound 29

(197) ##STR00195##

(198) Synthetic Route:

(199) ##STR00196##

Step 1: Synthesis of Compound 29

(200) m-Chloroperoxybenzoic acid (117.40 mg, 578.28 mol, purity: 85%) was added into the 24-A (206.69 mg, 578.28 mol) in toluene (15.00 mL) solution, the mixture was stirred at 20-25 C. for 1 hour and then added I2 (86.65 mg, 334.06 mol) and N,N-diisopropylethylamine (224.21 mg, 1.73 mmol, 302.99 L). After further stirring for 14 hours, the product was concentrated to give the crude compound, the crude compound was purified by preparative HPLC (neutral condition) to give the compound 29. .sup.1H NMR (400 MHz, DMSO-d) 61.47 (s, 6H) 1.47-1.50 (m, 4H) 1.54 (br s, 4H) 2.17 (s, 3H) 2.30 (br s, 4H) 3.09 (br s, 4H) 4.69 (br d, J=5.02 Hz, 2H) 4.83 (br d, J=17.07 Hz, 1H) 5.00 (br d, J=10.04 Hz, 1H) 5.34 (s, 1H) 5.60-5.79 (m, 1H) 6.92 (br d, J=8.53 Hz, 2H) 7.50-7.67 (m, 3H) 7.76 (br d, J=7.53 Hz, 1H) 8.06 (br s, 1H) 8.83 (s, 1H)

(201) MS m/z: 569.3[M+H].sup.+

Embodiment 30: Compound 30

(202) ##STR00197##

(203) Synthetic Route:

(204) ##STR00198##

Step 1: Synthesis of Compound 30-A

(205) Compound triethylamine (147.00 mg, 1.45 mmol, 201.37 L) was added into the compound I2-E (80.00 mg, 290.54 mol) in dichloromethane (2.00 mL) solution at 0 C., and then slowly added dropwise acetylchloride (45.61 mg, 581.08 mol, 41.46 L). The reaction mixture was stirred at 30 C. for 0.5 hours. Water (10 mL) was added into the reaction mixture, then extracted with dichloromethane (10 mL3) and washed by saturated brine (20 mL), dried over anhydrous sodium sulfate, then filtered, the filtrate was evaporated to give the crude product. The crude product was purified by column chromatography (PE/EtOAc=1/1-0/1) to give the compound 30-A. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.55 (q, J=5.68 Hz, 4H) 1.65-1.71 (m, 4H) 2.10 (s, 3H) 3.41-3.47 (m, 6H) 3.56-3.65 (m, 2H) 6.80 (d, J=8.46 Hz, 2H) 8.12 (d, J=9.54 Hz, 2H)

(206) MS m/z: 318.1 [M+H].sup.+

Step 2: Synthesis of Compound 30-B

(207) According to the method for preparing the compound 37-B, and started with the compound 30-A, the compound 30-B was obtained.

(208) MS m/z: 288.1 [M+H].sup.+

Step 3: Synthesis of Compound 30

(209) According to the method for preparing the compound 22, and started with the compound 30-B, the compound 30 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.52 (s, 6H) 1.64 (br d, J=5.52 Hz, 8H) 2.04 (s, 3H) 3.10 (t, J=5.52 Hz, 4H) 3.35-3.40 (m, 2H) 3.51-3.56 (m, 2H) 4.67 (d, J=6.02 Hz, 2H) 4.87 (dd, J=17.08, 1.00 Hz, 1H) 4.94-5.06 (m, 1H) 5.63 (ddt, J=16.82, 10.28, 6.28, 6.28 Hz, 1H) 6.87 (d, J=9.04 Hz, 2H) 7.27 (d, J=7.54 Hz, 1H) 7.40 (br d, J=8.54 Hz, 2H) 7.69 (d, J=8.04 Hz, 1H) 7.79 (t, J=8.04 Hz, 1H) 8.76 (s, 1H)

(210) MS m/z: 597.1 [M+H].sup.+

Embodiment 31: Compound 31

(211) ##STR00199##

(212) Synthetic Route:

(213) ##STR00200##

Step 1: Synthesis of Compound 31-A

(214) According to the method for preparing the compound 30-A, and acetylchloride was replaced by methyl chloroformate, the compound 31-A was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.47-1.56 (m, 4H) 1.61-1.69 (m, 4H) 3.40-3.52 (m, 8H) 3.64-3.75 (m, 3H) 6.80 (d, J=8.50 Hz, 2H) 8.09-8.14 (m, 2H)

(215) MS m/z: 334.1 [M+H].sup.+

Step 2: Synthesis of Compound 31-B

(216) According to the method for preparing the compound 37-B, and started with the compound 31-A, the compound 31-B was obtained.

(217) MS m/z: 304.1 [M+H].sup.+

Step 3: Synthesis of Compound 31

(218) According to the method for preparing the compound 22, and started with the compound 31-B, the compound 31 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.52 (s, 6H) 1.57-1.66 (m, 8H) 3.07-3.12 (m, 4H) 3.40 (br s, 4H) 3.63 (s, 3H) 3.90 (br s, 1H) 4.61-4.70 (m, 2H) 4.87 (dd, J=17.08, 1.00 Hz, 1H) 4.95-5.00 (m, 1H) 5.63 (ddt, J=17.00, 10.48, 6.08, 6.08 Hz, 1H) 6.86 (d, J=9.04 Hz, 2H) 7.27 (d, J=7.54 Hz, 1H) 7.39 (br d, J=8.54 Hz, 2H) 7.69 (d, J=8.04 Hz, 1H) 7.79 (t, J=7.78 Hz, 1H) 8.75 (s, 1H)

(219) MS m/z: 613.0 [M+H].sup.+

Embodiment 32: Compound 32

(220) ##STR00201##

(221) Synthetic Route:

(222) ##STR00202##

Step 1: Synthesis of Compound 32-A

(223) Formaldehyde solution (27.25 g, 335.75 mmol, 25.00 mL, 37% purity), Sodium triacetoxyborohydride (17.52 g, 82.66 mmol) and glacial acetic acid (7.45 g, 123.99 mmol, 7.10 mL) were added separately into the compound I2-A (10.10 g, 41.33 mmol) in methanol (150.00 mL) solution, and was stirred at 20-25 C. for 2 hours, the solvent was concentrated, 10% NaOH solution was used to adjust the pH to around 8, then extracted by dichloromethane 800 mL (150 mL6), the organic phase was washed by saturated brine 300 mL, dried over anhydrous sodium sulfate, then filtered and concentrated to give the compound 32-A. MS m/z: 259.1 [M+H].sup.+

Step 2: Synthesis of Compound 32-B

(224) Under argon atmosphere, acetic acid (7.88 g, 131.19 mmol, 7.50 mL) and wet palladium hydroxide/carbon (600.00 mg, 20% purity) were added into the compound 32-A (7.50 g, 29.02 mmol) in ethanol (100.00 mL) solution, after replacing with hydrogen for 3 times, the mixture was stirred at 50 C. and hydrogen (50 psi) for 18 hours, the product was purified by diatomite and then filtered and concentrated to give the crude product compound 32-B. MS m/z: 169.0 [M+H].sup.+

Step 3: Synthesis of Compound 32-C

(225) Triethylamine (361.25 mg, 3.57 mmol, 494.86 L) was added into the compound 32-B (200.00 mg, 1.19 mmol) and 5-chloro-2-nitro-pyridine (188.66 mg, 1.19 mmol) in 5 mL dimethyl sulfoxide solution, and was stirred at 90 C. for 12 hours, diluted by 10 mL 10% hydrochloric acid solution, by EtOAc 60 mL (20 mL3), the aqueous phase was adjusted by 10% sodium hydroxide to pH around 10, EtOAc 120 mL (40 mL3) was added. The organic phase washed by saturated brine 70 mL, dried over anhydrous sodium sulfate, then filtered and concentrated to give the compound 32-C. MS m/z: 291.1 [M+H].sup.+

Step 4: Synthesis of Compound 32-D

(226) Under argon atmosphere, 10% wet palladium carbon (25 mg) was added into the compound 32-C (200.00 mg, 688.80 mmol) in ethanol (20 mL) solution, after replacing with hydrogen 3 times, the mixture was stirred at 20-25 C. under hydrogen pressure (15 psi) for 16 hours, then filtered with diatomite, evaporated to give the crude product compound 32-D (brown oil, 180 mg). MS m/z: 261.3 [M+H].sup.+

Step 5: Synthesis of Compound 32

(227) According to the method for preparing the compound 5, and started with the compound 32-D, crude product of the compound 32 was obtained, the product was purified by preparative HPLC (neutral condition) to give the compound 32. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.85 (s, 1H), 8.11 (d, J=9.0 Hz, 1H), 7.96 (d, J=2.5 Hz, 1H), 7.81-7.87 (m, 1H), 7.65 (d, J=8.0 Hz, 1H), 7.31 (d, J=7.5 Hz, 1H), 7.23 (dd, J=9.3, 2.8 Hz, 1H), 5.58-5.70 (m, 1H), 4.99 (d, J=10.0 Hz, 1H), 4.87 (d, J=17.1 Hz, 1H), 4.66 (d, J=6.0 Hz, 2H), 3.89 (br s, 1H), 3.03-3.12 (m, 4H), 2.44 (br s, 4H), 2.30 (s, 3H), 1.62 (br s, 8H), 1.53 (s, 6H)

(228) MS m/z: 570.2[M+1].sup.+

Embodiment 33: Compound 33

(229) ##STR00203##

(230) Synthetic Route:

(231) ##STR00204##

Step 1: Synthesis of Compound 33-A

(232) Potassium carbonate (246.39 mg, 1.78 mmol) was added into the compound 32-B (100 mg, 594.25 mol) and 2,4-difluoro-nitrobenzene (435 mg, 2.73 mmol) in 3 mL dimethyl sulfoxide solution, and stirred at 90 C. for 12 hours, then diluted with 10 mL 10% hydrochloric acid solution, and washed by EtOAc 60 mL (20 mL3), the aqueous phase was adjusted by 10% sodium hydroxide to pH around 10, then extracted with EtOAc 120 mL (40 mL3). The organic phase was washed by saturated brine 60 mL, dried over anhydrous sodium sulfate, then filtered and concentrated to give the crude product compound 33-A. MS m/z: 308.1 [M+H].sup.+

Step 2: Synthesis of Compound 33-B

(233) According to the method for preparing the compound 32-D, and started with the compound 33-A, crude product of the compound 33-B was obtained. MS m/z: 277.9 [M+H].sup.+

Step 3: Synthesis of Compound 33

(234) According to the method for preparing compound 5, and started with the compound 33-B, crude product of the compound 33 was obtained, the product was purified by preparative HPLC (neutral condition) to give the compound 33. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.78 (s, 1H), 8.04 (s, 1H), 7.77-7.82 (m, 1H), 7.66 (d, J=8.0 Hz, 1H), 7.28 (d, J=7.5 Hz, 1H), 6.60-6.65 (m, 2H), 5.59-5.67 (m, 1H), 4.98 (d, J=9.5 Hz, 1H), 4.87 (d, J=17.1 Hz, 1H), 4.83-4.84 (m, 1H), 4.67 (d, J=6.0 Hz, 2H), 3.06-3.10 (m, 4H), 2.47 (br s, 4H), 2.32 (s, 3H), 1.58 (br s, 8H), 1.52 (s, 6H)

(235) MS m/z: 587.1[M+1].sup.+

Embodiment 34: Compound 34

(236) ##STR00205##

(237) Synthetic Route:

(238) ##STR00206##

Step 1: Synthesis of Compound 34-A

(239) Triethylamine (361.25 mg, 3.57 mmol, 494.86 L) was added into the compound 32-B (200.00 mg, 1.19 mmol) and 1,2-difluoro-4-nitrobenzene (350 mg, 2.20 mmol) in 10 mL methanol solution, and was stirred at 20-25 C. for 13 hours, diluted by 10 mL 10% hydrochloric acid solution, EtOAc 60 mL (20 mL3) was added, the aqueous phase was adjusted by 10% sodium hydroxide to pH around 10, EtOAc 120 mL (40 mL3) was used. The organic phase was washed by saturated brine 70 mL, dried over anhydrous sodium sulfate, then filtered and concentrated to give the crude product compound 34-A. MS m/z: 308.1 [M+H].sup.+

Step 2: Synthesis of Compound 34-B

(240) According to the method for preparing the compound 32-D, and started with the compound 34-A, crude product of the compound 34-B was obtained. MS m/z: 277.9 [M+H].sup.+

Step 3: Synthesis of Compound 34

(241) According to the method for preparing the compound 5, and started with the compound 34-B, crude product of the compound 34 was obtained, the product was purified by preparative HPLC (neutral condition) to give the compound 34. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.78 (s, 1H), 7.84-7.90 (m, 1H), 7.72 (d, J=8.0 Hz, 1H), 7.39-7.52 (m, 1H), 7.45 (br s, 1H), 7.31 (d, J=7.5 Hz, 1H), 6.90-6.95 (m, 1H), 6.83-6.89 (m, 1H), 5.59-5.69 (m, 1H), 4.96-5.01 (m, 1H), 4.87 (dd, J=17.1, 1.0 Hz, 1H), 4.69 (d, J=6.0 Hz, 2H), 2.92-2.97 (m, 4H), 2.46 (br s, 4H), 2.31 (s, 3H), 1.61 (br d, J=6.0 Hz, 8H), 1.52 (s, 6H)

(242) MS m/z: 587.1 [M/2+1].sup.+

Embodiment 35: Compound 35

(243) ##STR00207##

(244) Synthetic Route:

(245) ##STR00208##

Step 1: Synthesis of Compound 35-A

(246) According to the method for preparing the compound 33-A, 2,4-difluoro-nitrobenzene was replaced by 4-fluoro-2-methyl-nitrobenzene, the compound 35-A was obtained. MS m/z: 304.1 [M+H].sup.+

Step 2: Synthesis of Compound 35-B

(247) According to the method for preparing the compound 32-D, and started with the compound 35-A, the compound 35-B was obtained. MS m/z: 274.1 [M+H].sup.+

Step 3: Synthesis of Compound 35

(248) According to the method for preparing the compound 5, and started with the compound 35-B, crude product of the compound 35 was obtained, the product was purified by preparative HPLC (neutral condition) to give the compound 35. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.75 (s, 1H), 7.72 (br s, 1H), 7.63 (br d, J=8.0 Hz, 1H), 7.24 (d, J=7.5 Hz, 1H), 7.02 (br s, 1H), 6.70-6.75 (m, 2H), 5.57-5.67 (m, 1H), 4.97 (d, J=10.3 Hz, 1H), 4.87 (d, J=17.1 Hz, 1H), 4.67 (br d, J=6.0 Hz, 2H), 3.86 (br s, 1H), 3.07-3.13 (m, 4H), 2.87 (br s, 4H), 2.59 (s, 3H), 2.18 (s, 3H), 1.81 (br s, 3H), 1.72-1.78 (m, 3H), 1.63-1.69 (m, 6H)

(249) MS m/z: 583.1[M+1].sup.+

Embodiment 36: Compound 36

(250) ##STR00209##

(251) Synthetic Route:

(252) ##STR00210##

Step 1: Synthesis of Compound 36-A

(253) According to the method for preparing the compound 33-A, 2,4-difluoro-nitrobenzene was replaced by 1-fluoro-2-methyl-4-nitrobenzene, the compound 36-A was obtained. MS m/z: 304.2 [M+H].sup.+.

Step 2: Synthesis of Compound 36-B

(254) According to the method for preparing the compound 32-D, and started with the compound 36-A, the compound 36-B was obtained. MS m/z: 274.3 [M+H].sup.+.

Step 3: Synthesis of Compound 36

(255) According to the method for preparing the compound 22, and started with the compound 36-B, the compound 36 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.52 (s, 6H) 1.61-1.65 (m, 4H) 1.78 (br t, J=5.28 Hz, 4H) 2.24 (s, 3H) 2.51 (s, 3H) 2.74-2.82 (m, 8H) 4.68 (br d, J=6.54 Hz, 2H) 4.86 (d, J=18.08 Hz, 1H) 4.97 (d, J=10.54 Hz, 1H) 5.63 (ddt, J=16.82, 10.42, 6.22, 6.22 Hz, 1H) 6.93 (d, J=8.54 Hz, 1H) 7.24 (dd, J=8.54, 2.51 Hz, 1H) 7.29 (d, J=7.54 Hz, 1H) 7.43 (br s, 1H) 7.70-7.75 (m, 1H) 7.76-7.86 (m, 1H) 8.77 (s, 1H)

(256) MS m/z: 583.1 [M+H].sup.+

Embodiment 37: Compound 37

(257) ##STR00211##

(258) Synthetic Route:

(259) ##STR00212##

Step 1: Synthesis of Compound 37-A

(260) Potassium carbonate (323.41 mg, 2.34 mmol) and compound 32-B (334.71 mg, 1.99 mmol) were added into the compound 5-fluoro-2-nitroanisole (200.00 mg, 1.17 mmol) in dimethyl sulfoxide (7.00 mL) solution. The reaction mixture was stirred at 100 C. for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent and give the crude product. The crude compound preparative HPLC (neutral) to give the compound 37-A. MS m/z: 320.1 [M+H].sup.+

Step 2: Synthesis of Compound 37-B

(261) Pd/C (39.50 mg, 33.50 mol, 10% purity) was added into the compound 37-A (107.00 mg, 335.00 mol) in THF (10.00 mL) solution. The reaction mixture was stirred at 30 C. for 12 hours under hydrogen pressure (15 Psi). The reaction mixture was filtered, the filtrate was concentrated and evaporated to give the compound 37-B. MS m/z: 290.3 [M+H].sup.+

Step 3: Synthesis of Compound 37

(262) According to the method for preparing the compound 22, and started with the compound 37-B, the compound 37 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.52 (s, 6H) 1.57-1.63 (m, 8H) 2.24 (s, 3H) 2.34 (br s, 4H) 3.05-3.09 (m, 4H) 3.81 (s, 3H) 3.92 (br s, 1H) 4.67 (d, J=6.02 Hz, 2H) 4.87 (d, J=17.08 Hz, 1H) 4.97 (d, J=10.04 Hz, 1H) 5.64 (ddt, J=16.82, 10.42, 6.21, 6.21 Hz, 1H) 6.45-6.49 (m, 2H) 7.28 (d, J=7.54 Hz, 1H) 7.73 (d, J=8.04 Hz, 1H) 7.80-7.86 (m, 1H) 8.10 (br d, J=7.54 Hz, 1H) 8.77 (s, 1H)

(263) MS m/z: 599.1 [M+H].sup.+

Embodiment 38: Compound 38

(264) ##STR00213##

(265) Synthetic Route:

(266) ##STR00214##

Step 1: Synthesis of Compound 38-A

(267) Triethylamine (266.09 mg, 2.63 mmol, 364.51 L) was added into the compound 32-B (221.25 mg, 1.31 mmol) and 1-fluoro-methoxyl-nitro-benzene (150 mg, 876.53 mol) in 10 mL methanol solution, and was stirred at 60 C. for 32 hours, the mixture was then poured into 20 mL water, and was stirred for 30 min, then filtered, the filtrate was adjusted by 2 mol/L hydrochloric acid to pH=1-2, after washing by EtOAc (30 mL2), 4 mol/L sodium hydroxide was used to adjust the pH=10-11, then extracted by EtOAc (40 mL3), dried over anhydrous sodium sulfate, then filtered and concentrated to give the crude product compound 38-A. MS m/z: 319.9 [M+H].sup.+

Step 2: Synthesis of Compound 38-B

(268) According to the method for preparing the compound 32-D, and started with the compound 38-A, the compound 38-B was obtained. MS m/z: 290.2 [M+H].sup.+

Step 3: Synthesis of Compound 38

(269) According to the method for preparing the compound 22, and started with the compound 38-B, the compound 38 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.60 (s, 6H) 1.64-1.72 (m, 8H) 2.31-2.36 (m, 3H) 2.46 (br s, 4H) 2.94-3.03 (m, 4H) 3.82 (s, 3H) 4.75 (d, J=6.02 Hz, 2H) 4.95 (d, J=16.06 Hz, 1H) 5.06 (d, J=10.04 Hz, 1H) 5.66-5.78 (m, 1H) 6.94 (d, J=8.54 Hz, 1H) 7.11 (br s, 2H) 7.35-7.46 (m, 1H) 7.76 (d, J=8.04 Hz, 1H) 7.86 (t, J=7.78 Hz, 1H) 8.87 (s, 1H)

(270) MS m/z: 599.1 [M+H].sup.+

Embodiment 39: Compound 39

(271) ##STR00215##

(272) Synthetic Route:

(273) ##STR00216##

Step 1: Synthesis of Compound 39-A

(274) Diisopropylethylamine (3.58 g, 27.67 mmol) was added into 4-chloro-2-methylthiopyrimidin-5-ethyl carboxylate (2.80 g, 12.03 mmol) and 2-chloro phenyl hydrazine (2.48 g, 13.83 mmol) in 90 mL THF solution, then heated to 80 C. and stirred for 16 hours. The reaction was monitored to be complete by LCMS. After concentrated under reduced pressure to remove the solvent, water 50 mL was added, and extracted by 20 mL EtOAc for 3 times, the combined EtOAc phase were washed by brine 20 mL, dried over anhydrous sodium sulfate, concentrated under reduced pressure to give the compound 39-A. .sup.1H NMR (400 MHz, CDCl.sub.3) =9.65 (br d, J=3.8 Hz, 1H), 8.71-8.65 (m, 1H), 7.30 (dd, J=1.2, 8.0 Hz, 1H), 7.15-7.08 (m, 1H), 6.92 (dd, J=1.6, 8.0 Hz, 1H), 6.84 (dt, J=1.6, 8.0 Hz, 1H), 6.78 (d, J=4.0 Hz, 1H), 4.40 (q, J=7.2 Hz, 2H), 2.25 (s, 3H), 1.42 (t, J=7.0 Hz, 3H)

Step 2: Synthesis of Compound 39-B

(275) Sodium hydroxide (5 M, 15 mL) was added into compound 39-A (4.00 g, 11.81 mmol) in methanol (30 mL) and THF (30 mL) solution, then stirred at 20-25 C. for 3 hours. The reaction was monitored to be complete by LCMS. The reaction mixture was concentrated under reduced pressure, then added 150 mL water, 6 N hydrochloric acid was used to adjust pH=3, then filtered to give a solid. The solid was washed by water 100 mL3, dried to give the compound 39-B. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =9.79 (s, 1H), 8.60 (s, 1H), 7.80 (s, 1H), 7.33 (dd, J=1.2, 7.8 Hz, 1H), 7.16-7.09 (m, 1H), 6.85-6.72 (m, 2H), 2.17 (s, 3H)

Step 3: Synthesis of Compound 39-C

(276) Toluene 25 mL and dichlorosulfoxide (8.20 g, 68.94 mmol) were added into compound 39-B (1.00 g, 3.22 mmol), the mixture was heated to 110 C. and stirred for 1 hour. The reaction was monitored to be complete by LCMS. After cooled down to r.t., the mixture was poured into the mixture of 200 mL ice and saturated sodium bicarbonate while stirring, then extracted by EtOAc (50 mL3), the combined organic phases was washed by brine 50 mL, dried over anhydrous sodium sulfate, concentrated under reduced pressure to give the compound 39-C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =8.81-8.65 (m, 1H), 7.72-7.66 (m, 1H), 7.62-7.50 (m, 4H), 2.63 (s, 3H)

Step 4: Synthesis of Compound 39-D

(277) Sodium carbonate (220.13 mg, 2.08 mmol) and 3-bromopropene (413.27 mg, 3.42 mmol) were added into compound 39-C (200.00 mg, 683.20 mol) in 5 mL acetonitrile solution, then the mixture was heated to 85 C. and stirred for 1.5 hours. The reaction was monitored to be complete by LCMS. Water 20 mL was added into the mixture, then extracted by EtOAc (6 mL3), the combined organic phases were washed by brine 6 mL, dried over anhydrous sodium sulfate, concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (PE/EtOAc=4/1-0/1) to give the compound 39-D. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =8.97 (s, 1H), 7.71-7.67 (m, 1H), 7.60-7.50 (m, 3H), 5.64 (m, 1H), 5.09 (dd, J=1.6, 10.4 Hz, 1H), 4.98 (dd, J=1.6, 17.2 Hz, 1H), 4.37 (t, J=6.0 Hz, 2H), 2.58 (s, 3H)

Step 5: Synthesis of Compound 39

(278) m-Chloroperoxybenzoic acid (51.33 mg, 297.46 mol) was added into compound 39-D (90.00 mg, 270.42 mol) in 2 mL dichloromethane solution. The mixture was stirred at 25 C. for 60 min. Diisopropylethylamine (209.70 mg, 1.62 mmol) and 12 (73.65 mg, 283.94 mol) were then added, the mixture was stirred at 25 C. for 16 hours. The reaction was monitored to be complete by LCMS. The reaction mixture was diluted by 20 mL dichloromethane, and washed by saturated sodium sulfite solution 10 mL and saturated sodium bicarbonate 10 mL, dried over anhydrous sodium sulfate, concentrated under reduced pressure to give the residue. The residue was separated by preparative HPLC to give the compound 39. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.81 (s, 1H), 7.58-7.47 (m, 3H), 7.44-7.34 (m, 3H), 6.96 (d, J=9.0 Hz, 2H), 5.68 (m, 1H), 5.18-5.00 (m, 2H), 4.39-4.19 (m, 2H), 3.19-3.08 (m, 4H), 2.40 (br s, 4H), 2.30 (s, 3H), 1.68-1.62 (m, 4H), 1.60-1.57 (m, 4H)

(279) MS m/z: 554.1 [M+H].sup.+

Embodiment 40: Compound 40

(280) ##STR00217##

(281) Synthetic Route:

(282) ##STR00218##

Step 1: Synthesis of Compound 40-A

(283) n-Butyllithium (2.5 M, 1.86 mL) was added into 2,6-dibromopyridine (1.00 g, 4.22 mmol) in 30 mL dichloromethane suspension at 60 C. The reaction mixture was stirred for 15 min, cyclobutanone (355.05 mg, 5.06 mmol) was added at one time into the reaction mixture at 60 C. The reaction mixture was stirred at 60 C. for 30 min. The reaction was monitored to be complete by LCMS. The reaction mixture was poured into saturated ammonium chloride, the organic phase was separated and washed by brine 10 mL, dried over anhydrous sodium sulfate, concentrated under reduced pressure to give a dark red oil. The residue was purified by column chromatography (PE/EtOAc=4/1) to give the compound 40-A. The product was confirmed by .sup.1H NMR (CDCl.sub.3).

(284) .sup.1H NMR (400 MHz, CDCl.sub.3) =7.62-7.57 (m, 1H), 7.53-7.50 (dd, J=0.8, 7.6 Hz, 1H), 7.39 (dd, J=0.8, 7.6 Hz, 1H), 4.30 (br, 1H), 2.55-2.45 (m, 4H), 2.12-2.00 (m, 1H), 1.91-1.78 (m, 1H)

Step 2: Synthesis of Compound 40-B

(285) According to the method for preparing the compound 3-A and 2-bromopyridine was replaced by 40-A, the compound 40-B was obtained. MS m z: 370.1 [M+H].sup.+

Step 3: Synthesis of Compound 23

(286) According to the method for preparing the compound 16, and started with the compound 40-B, crude product of the compound 40 was obtained, the product was purified by preparative separation (neutral condition) to give the compound 23. .sup.1H NMR (CDCl.sub.3, 400 MHz): =8.76 (s, 1H), 7.80-7.86 (m, 1H), 7.70 (d, J=7.8 Hz, 1H), 7.36-7.44 (m, 3H), 6.86 (d, J=9.0 Hz, 2H), 5.64 (ddt, J=16.8, 10.4, 6.2 Hz, 1H), 4.98 (dd, J=10.3, 1.0 Hz, 1H), 4.88 (dd, J=17.1, 1.3 Hz, 1H), 4.66 (br d, J=6.3 Hz, 2H), 4.00 (s, 1H), 3.04-3.12 (m, 4H), 2.46 (br t, J=7.9 Hz, 8H), 2.31 (s, 3H), 1.99-2.06 (m, 1H), 1.77-1.89 (m, 1H), 1.61 ppm (br s, 8H).

(287) MS m/z: 581.1[M+1].sup.+

Embodiment 41: Compound 41

(288) ##STR00219##

(289) Synthetic Route:

(290) ##STR00220##

Step 1: Synthesis of Compound 41-A

(291) Compound 2-bromo-6-acetyl pyridine (500.00 mg, 2.50 mmol) was added into sodium acetate (205.04 mg, 2.50 mmol) in dimethyl sulfoxide (3.00 mL) suspension. Trifluoromethyl trimethylsilane (1.42 g, 10.00 mmol) in dimethyl sulfoxide (0.50 mL) solution was added dropwise at 10-25 C., after the completion of the adding, the reaction mixture was reacted at 10-25 C. for 12 hours. Water (4.00 mL) was added to quench the reaction, the reaction flask was cooled down in an ice bath, the inner temperature was 10-25 C. After extraction by EtOAc (9.00 mL3), the organic phases were combined, the organic phases were washed by saturated sodium bicarbonate (10.00 mL), then washed by saturated brine (10 mL), dried over anhydrous sodium sulfate. The desiccant was filtered off, the filtrate was concentrated under reduced pressure to remove the solvent and give the crude product. The crude product was purified by column chromatography (PE/EtOAc=15/1) to give 41-A. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.57 (s, 3H) 5.39 (s, 1H) 7.33 (d, J=7.54 Hz, 1H) 7.40 (d, J=8.04 Hz, 1H) 7.52 (t, J=7.78 Hz, 1H)

(292) MS m/z: 271.8 [M+H].sup.+

Step 2: Synthesis of Compound 41-B

(293) According to the method for preparing the compound 27-A, the 2-bromo-6-fluoropyridine was replaced by 41-A, crude product of the compound 41-B was obtained, the crude product was purified by column chromatography (PE/EtOAc=3/1, 0/1) to give the compound 41-B. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.59 (s, 3H) 2.41 (s, 3H) 4.38-4.47 (m, 1H) 4.62-4.79 (m, 2H) 4.86-4.91 (m, 1H) 5.12 (s, 1H) 5.50 (ddt, J=16.68, 10.54, 6.22, 6.22 Hz, 1H) 7.30 (d, J=7.54 Hz, 1H) 7.75-7.79 (m, 1H) 7.81-7.87 (m, 1H) 8.77 (s, 1H)

Step 3: Synthesis of Compound 41

(294) According to the method for preparing the compound 22, and started with the compound 41-B, the compound 41 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.58-1.67 (m, 8H) 1.70 (s, 3H) 2.35 (s, 3H) 2.51 (br s, 4H) 3.05-3.11 (m, 4H) 4.50 (dd, J=15.56, 6.54 Hz, 1H) 4.71 (dd, J=15.56, 5.52 Hz, 1H) 4.86 (d, J=17.08 Hz, 1H) 4.98 (d, J=10.04 Hz, 1H) 5.56-5.67 (m, 1H) 6.86 (d, J=9.04 Hz, 2H) 7.36 (br d, J=8.54 Hz, 3H) 7.86-7.90 (m, 2H) 8.77 (s, 1H)

(295) MS m/z: 623.1 [M+H].sup.+

Embodiment 42: Compound 42

(296) ##STR00221##

(297) Synthetic Route:

(298) ##STR00222##

Step 1: Synthesis of Compound 42-A

(299) n-Butyllithium (2.5 M, 1.86 mL) was added into 2,6-dibromopyridine (1.00 g, 4.22 mmol) in 30 mL dichloromethane suspension at 60 C. The reaction mixture was stirred for 15 min, then 3-carbonyl oxetane (304.10 mg, 4.22 mmol) was added for one-time into the reaction mixture at 60 C. The reaction mixture was stirred for further 60 min at 60 C. The reaction was monitored to be complete by thin layer chromatography. The reaction mixture was poured into saturated ammonium chloride, the organic phases were separated, and washed by brine 10 mL, dried over anhydrous sodium sulfate, concentrated under reduced pressure to give yellow solid. The residue was purified by column chromatography (PE/EtOAc=2/1) to give the compound 42-A. .sup.1H NMR (400 MHz, CDCl.sub.3) =7.86 (dd, J=0.8, 7.8 Hz, 1H), 7.67 (t, J=7.8 Hz, 1H), 7.44 (d, J=7.8 Hz, 1H), 5.20 (s, 1H), 5.04-4.96 (m, 2H), 4.69-4.59 (m, 2H)

Step 2: Synthesis of Compound 42-B

(300) N,N-dimethylethylenediamine (43.62 mg, 494.89 mol) was added into the mixture of I1 (100.00 mg, 449.90 mol), cuprous iodide (85.68 mg, 449.90 mol), compound 42-A (103.50 mg, 449.90 mol) and potassium carbonate (87.05 mg, 629.86 mol) in 5 mL dioxane, under nitrogen atmosphere, the mixture was heated to 95 C. and stirred for 2 hours. The reaction was monitored to be complete by LCMS. The reaction mixture was cooled down, then 30 mL ammonia was added, and then extracted with EtOAc (10 mL2), washed by saturated brine, dried over anhydrous magnesium sulfate, concentrated under reduced pressure to give 42-B. MS m/z: 372.1 [M+H].sup.+

Step 3: Synthesis of Compound 42

(301) m-Chloroperoxybenzoic acid (110.69 mg, 545.22 mol, 85%) was added into compound 42-B (150.00 mg, 403.87 mol) in 20 mL toluene solution at 25-30 C. The mixture was stirred for 30 min at 25-30 C. Diisopropylethylamine (143.54 8 mg, 1.11 mmol) and 12 (104.76 mg, 403.87 mol) was added into the mixture under 30 C., and the mixture was stirred at 30-32 C. for 15 hours. The reaction was monitored to be complete by LCMS. The reaction mixture was diluted by 40 mL EtOAc, then washed sequentially by saturated sodium sulfite solution 20 mL, saturated sodium bicarbonate 20 mL and brine 20 mL, dried over anhydrous sodium sulfate, concentrated under reduced pressure to give the residue. The residue was separated by preparative HPLC, and then purified by thin layer chromatography (dichloromethane/methanol=10/1) to give the compound 42. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.76 (s, 1H), 8.00-7.93 (m, 1H), 7.80 (dd, J=7.7, 13.7 Hz, 2H), 7.36 (br d, J=8.3 Hz, 3H), 6.86 (d, J=9.0 Hz, 2H), 5.70-5.58 (m, 1H), 5.15 (br s, 1H), 5.04-5.00 (m, 3H), 4.90 (d, J=17.2 Hz, 1H), 4.70 (d, J=7.2 Hz, 2H), 4.57 (d, J=6.0 Hz, 2H), 3.13-3.05 (m, 4H), 2.33 (br s, 4H), 2.23 (s, 3H), 1.64-1.55 (m, 8H)

(302) MS m/z: 583.1 [M+H].sup.+

Embodiment 43: Compound 43

(303) ##STR00223##

(304) Synthetic Route:

(305) ##STR00224##

Step 1: Synthesis of Compound 43-A

(306) At 10 C. and under nitrogen atmosphere, diethylaminosulfur trifluoride (70.06 mg, 434.66 mol) was added dropwise rapidly into the compound 42-A (50.00 mg, 217.33 mol) in 2 mL dichloromethane solution. Then the mixture was stirred at 10 C. for 30 min under nitrogen atmosphere. The reaction was monitored to be complete by thin layer chromatography. The reaction mixture was quenched by 10 mL saturated sodium bicarbonate, and then extracted by dichloromethane (10 mL2), the combined organic phases were dried over anhydrous sodium sulfate, then concentrated to give the compound 43-A. The product was used directly in the next step without further purification. MS m/z: 231.8 [M+H].sup.+, 233.9 [M+H].sup.+

Step 2: Synthesis of Compound 43-B

(307) According to the method for preparing the compound 27-A, and 2-bromo-6-fluoropyridine was replaced by 43-A, the compound 43-B was obtained. MS m/z: 374.1 [M+H].sup.+

Step 3: Synthesis of Compound 43

(308) According to the method for preparing the compound 22, and started with the compound 43-B, the compound 43 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.60-1.71 (m, 8H) 2.32 (s, 3H) 2.43 (br s, 4H) 3.15-3.20 (m, 4H) 4.90-4.94 (m, 2H) 5.01 (dd, J=14.32, 6.28 Hz, 2H) 5.07 (d, J=7.04 Hz, 2H) 5.11 (d, J=7.54 Hz, 1H) 5.17 (d, J=8.04 Hz, 1H) 5.73 (ddt, J=16.76, 10.36, 6.28, 6.28 Hz, 1H) 6.95 (d, J=9.04 Hz, 2H) 7.40-7.50 (m, 3H) 7.90 (t, J=7.84 Hz, 1H) 8.01 (d, J=8.54 Hz, 1H) 8.85 (s, 1H)

(309) MS m/z: 585.1 [M+H].sup.+

Embodiment 44: Compound 44

(310) ##STR00225##

(311) Synthetic Route:

(312) ##STR00226##

Step 1: Synthesis of Compound 44-A

(313) Potassium carbonate (402.57 mg, 2.91 mmol), cuprous iodide (401.98 mg, 2.11 mmol), N,N-dimethylethylenediamine (208.38 mg, 2.36 mmol, 254.12 L), and compound 2-pyrrolidone (250.00 mg, 2.94 mmol, 225.23 L, 1.39 eq) were added into compound 2,6-dibromopyridine (500.00 mg, 2.11 mmol) in toluene (15.00 mL) solution, the reaction mixture was stirred at 110 C. for 12 hours under nitrogen atmosphere. The reaction was cooled down to r.t., ammonia (30 mL) was added into the reaction mixture, extracted by EtOAc (20 mL3), the organic phases were combined, and washed by saturated brine (50 mL), dried over anhydrous sodium sulfate, the organic phase was concentrated under reduced pressure to give the crude product, which was purified by column chromatography (PE/EtOAc=5/1, 3/1) to give the compound 44-A. MS m/z: 240.8 [M+H].sup.+

Step 2: Synthesis of Compound 44-B

(314) According to the method for preparing the compound 27-A, and the 2-bromo-6-fluoropyridine was replaced by 44-A to give the compound 44-B.

(315) MS m/z: 383.1[M+H]+, 405.1 [M+Na].sup.+

Step 3: Synthesis of Compound 44

(316) According to the method for preparing the compound 22, and started with the compound 44-B, the compound 44 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.51-1.61 (m, 8H) 2.05-2.12 (m, 2H) 2.24 (s, 3H) 2.34 (br s, 4H) 2.62 (t, J=8.04 Hz, 2H) 3.06-3.10 (m, 4H) 4.02 (t, J=7.04 Hz, 2H) 4.67-4.72 (d, J=6.02 Hz, 2H) 4.88 (d, J=17.08 Hz, 1H) 4.97 (d, J=10.04 Hz, 1H) 5.61 (ddt, J=16.88, 10.36, 6.22, 6.22 Hz, 1H) 6.85 (d, J=9.04 Hz, 2H) 7.36 (br d, J=8.54 Hz, 2H) 7.52 (d, J=8.04 Hz, 1H) 7.77 (t, J=8.04 Hz, 1H) 8.26 (d, J=8.04 Hz, 1H) 8.74 (s, 1H)

(317) MS m/z: 594.1 [M+H].sup.+

Embodiment 45: Compound 45

(318) ##STR00227##

(319) Synthetic Route:

(320) ##STR00228##

Step 1: Synthesis of Compound 45-A

(321) Potassium phosphate (2.69 g, 12.66 mmol), potassium cyclopropyltrifluoroborate (624.68 mg, 4.22 mmol) and tetrakis(triphenylphosphine)palladium (487.80 mg, 422.00 mol) were added into compound 2,6-dibromopyridine (1.00 g, 4.22 mmol) in the mixture of toluene (30.00 mL) and water (3.00 mL), the reaction was stirred at 95 C. for 12 hours under nitrogen atmosphere. The reaction was cooled down to 25 C., then water (20 mL) was added, extracted by EtOAc (30 mL3), the organic phases were combined and washed by saturated brine (50 mL), dried over anhydrous sodium sulfate, then filtered, the filtrate was evaporated to give the crude product, the crude product was purified by column chromatography (PE/EtOAc=I/O) to give the compound 45-A. MS m/z: 199.8 [M+H].sup.+

Step 2: Synthesis of Compound 45-B

(322) According to the method for preparing the compound 27-A, and 2-bromo-6-fluoropyridine was replaced by 45-A, the compound 45-B was obtained. MS m/z: 340.0 [M+H].sup.+

Step 3: Synthesis of Compound 45

(323) According to the method for preparing the compound 22, and started with the compound 45-B, compound 45 was obtained. The method for synthesis of compound 45 was the same as the embodiment 22 used for synthesis of compound 22 except for the starting material, the compound 45 was thus obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) 0.98-1.07 (m, 4H) 1.58-1.76 (m, 8H) 1.99-2.11 (m, 2H) 2.35 (s, 3H) 2.37-2.53 (m, 4H) 3.12-3.16 (m, 2H) 3.45-3.55 (m, 1H) 4.76 (br d, J=6.54 Hz, 2H) 4.91 (d, J=17.08 Hz, 1H) 5.01 (d, J=10.04 Hz, 1H) 5.61-5.74 (m, 1H) 6.85-6.94 (m, 2H) 7.06-7.11 (m, 1H) 7.45 (br d, J=8.54 Hz, 2H) 7.56-7.62 (m, 1H) 7.64-7.72 (m, 1H) 8.81 (s, 1H)

(324) MS m/z: 551.1 [M+H].sup.+

Embodiment 46: Compound 46

(325) ##STR00229##

(326) Synthetic Route:

(327) ##STR00230##

Step 1: Synthesis of Compound 46-A

(328) N,N-dimethylethylenediamine (87.25 mg, 989.79 mol) was added into the mixture of I1 (200.00 mg, 899.81 mol), cuprous iodide (171.37 mg, 899.81 mol), 2-trifluoromethyl-6-bromopyridine (209.45 mg, 926.80 mol) and potassium carbonate (174.11 mg, 1.26 mol) in 3 mL dioxane, the mixture was heated to 95 C. and stirred for 1 hour under nitrogen atmosphere. The reaction was monitored to be complete by LCMS. The reaction mixture was cooled down, 30 mL ammonia was added, then extracted by EtOAc (10 mL2), then washed by saturated brine, dried over anhydrous magnesium sulfate. The filtrate was concentrated under reduced pressure to give the pale brown colloidal product, which was purified by column chromatography (PE/EtOAc=4/1-3/1) to give the compound 46-A. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.89 (s, 1H), 8.20 (d, J=8.5 Hz, 1H), 7.98 (t, J=8.0 Hz, 1H), 7.54 (d, J=7.8 Hz, 1H), 5.64-5.54 (m 1H), 4.94 (d, J=10.4 Hz, 1H), 4.90-4.83 (m, 3H), 2.54 (s, 3H)

Step 2: Synthesis of Compound 46

(329) At 30-40 C., m-chloroperoxybenzoic acid (52.23 mg, 297.46 mol, 85%) was added into compound 46-A (70.00 mg, 190.55 mol) in 5 mL toluene solution. The mixture was stirred at 25-30 C. for 30 min. Then diisopropylethylamine (73.88 mg, 571.65 mol) and 12 (49.43 mg, 190.55 mol) were added below 30 C., the mixture was stirred at 25-30 C. for 16 hours. The reaction was monitored to be complete by LCMS. The reaction mixture was diluted by 20 mL EtOAc, then washed sequentially by saturated sodium sulfite solution 10 mL, saturated sodium bicarbonate 10 mL and brine 10 mL, dried over anhydrous sodium sulfate, concentrated under reduced pressure to give the residue, which was separated by preparative HPLC to give the compound 46. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.77 (s, 1H), 8.16 (d, J=8.4 Hz, 1H), 7.89 (t, J=7.8 Hz, 1H), 7.48 (d, J=7.6 Hz, 1H), 7.35 (d, J=9.0 Hz, 2H), 6.87 (d, J=9.0 Hz, 2H), 5.63-5.56 (m, 1H), 4.96-4.79 (m, 4H), 3.15-3.06 (m, 4H), 2.35 (br s, 4H), 2.24 (s, 3H), 1.62-1.51 (m, 8H)

(330) MS m/z: 579.0 [M+H].sup.+

Embodiment 47: Compound 47

(331) ##STR00231##

(332) Synthetic Route:

(333) ##STR00232##

Step 1: Synthesis of Compound 47-1

(334) I1 (1.0 g, 4.5 mmol) was added into 1,4-dioxane (10 mL), and 2-bromo-6-methoxylpyridine (1.02 g, 5.4 mmol, 664 L), potassium carbonate (622 mg, 4.5 mmol), cuprous iodide (857 mg, 4.5 mmol) and N,N-dimethylethylenediamine (397 mg, 4.5 mmol, 490 L) were added while stirring, heated to 95 C. under nitrogen atmosphere, then reacted for 12 h. 100 mL ammonia was added into the reacted mixture, then extracted by 100 mL EA, the organic phase was washed by 100 mL brine, dried over anhydrous sodium sulfate, filtered and concentrated, the crude product was crystallized by EtOAc, then purified by silica gel chromatography (PE/EA=10/1 to 2/1), to give the compound 47-1. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.92 (s, 1H), 7.89 (t, J=8.0 Hz, 1H), 7.43 (d, J=7.2 Hz, 1H), 6.82 (d, J=8.0 Hz, 1H), 5.83-5.69 (m, 1H), 5.07 (d, J=10.4 Hz, 1H), 4.96 (br d, J=17.2 Hz, 1H), 4.84 (d, J=6.0 Hz, 2H), 3.93 (s, 3H), 2.56 (s, 3H)

Step 2: Synthesis of Compound 47

(335) m-CPBA (139.71 mg, 688.15 mol, 85% purity) was added into 47-1 (200 mg, 607 mol) in 1 mL DCM solution, and was stirred at 25 C. for 30 min. 12 (272.47 mg, 1.05 mmol) and DIPEA (215.8 mg, 1.67 mmol, 291.62 L) were added at 30 C. and stirred for 12 h. The reacted mixture was partitioned between 10 mL DCM and 10 mL, the organic phase was washed by 10 mL saturated sodium sulfite solution, 10 mL sodium carbonate solution and 10 mL sodium chloride solution, then dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated, the product was purified by preparative HPLC (chromatographic column: waters Xbridge 15025 mmmm 5 um; mobile phases: [water (0.225% FA)-ACN]; B %: 10%-30%, 12 min).) to give the compound 47. .sup.1H NMR (400 MHz, MeOD) 8.79 (s, 1H) 7.66-7.80 (m, 1H) 7.38-7.54 (m, 2H) 6.90 (d, J=8.8 Hz, 2H) 6.61-6.70 (m, 2H) 5.64-5.87 (m, 1H) 4.91-5.07 (m, 2H) 4.70-4.82 (m, 2H) 3.92 (s, 3H) 3.12-3.15 (br, 4H) 3.12-3.15 (m, 4H) 2.31-2.52 (m, 4H) 2.28 (s, 3H) 1.54-1.73 (m, 8H) MS m/z: 541.3 [M+H].sup.+

Embodiment 48: Compound 48

(336) ##STR00233##

(337) Synthetic Route:

(338) ##STR00234##

Step 1: Synthesis of Compound 48-1

(339) 2-Bromo-6-hydroxypyridine (1.00 g, 5.75 mmol) was dissolved in 10 mL in DMF, potassium carbonate (1.59 g, 11.50 mmol) was added at 0 C., and stirred for 30 min, then iodoisopropane (1.17 g, 6.90 mmol, 689.96 L) was added dropwise. The reaction mixture was then stirred at 0 C. for 30 min, then warmed to 20 C., then the mixture was stirred for 12 h. The reaction mixture was partitioned between and extracted by 30 mL water and 30 mL, the organic phase was washed by 30 mL brine, then dried, and the filtrate was concentrated to give 48-1. .sup.1H NMR (400 MHz, CDCl.sub.3) 7.38 (t, J=7.2 Hz, 1H) 7.00 (d, J=7.2 Hz, 1H) 6.61 (d, J=8.0 Hz, 1H) 5.26-5.432 (m, 1H) 1.35 (dd, J=6.0, 1.6 Hz, 6H)

Step 2: Synthesis of Compound 48-2

(340) I1 (980.00 mg, 4.41 mmol) was added into 1,4-dioxane (10.00 mL), and 2-bromo-6-isopropylpyridine (1.14 g, 5.29 mmol), potassium carbonate (835.02 mg, 6.04 mmol), cuprous iodide (839.70 mg, 4.41 mmol) and N,N-dimethylethylenediamine (427.62 mg, 4.85 mmol, 527.92 L) were added while stirring, heated to 95 C. under nitrogen atmosphere, then stirred for 12 h. 100 mL ammonia was added into the reacted mixture, the mixture was extracted by 100 mL EA, the organic phase was washed by 100 mL brine, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated to give a residue, which was then crystallized in EtOAc, then purified by silica gel chromatography (PE/EA=10/1-2/1) to give 48-2. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.90 (s, 1H) 7.71 (t, J=8.0 Hz, 1H) 7.31 (d, J=7.6 Hz, 1H) 6.63 (d, J=8.2 Hz, 1H) 5.54-5.82 (m, 1H) 5.12-5.32 (m, 1H) 4.90-5.24 (m, 2H) 4.78 (d, J=6.4 Hz, 2H) 2.55 (s, 3H) 1.32 (d, J=6.0 Hz, 6H)

Step 3: Synthesis of Compound 48

(341) Compound 48-2 (525.00 mg, 1.47 mmol) and m-CPBA (338.24 mg, 1.67 mmol) were added into 1 mL DCM, and stirred at 25 C. for 1 h. 12 (659.65 mg, 2.54 mmol) and DIPEA (522.45 mg, 4.04 mmol, 706.01 L) were added into the mixture, and stirred at 30 C. for 12 h. The reacted mixture was partitioned between 10 mL DCM and 10 mL, the organic phase was washed separately with 10 mL saturated sodium sulfite solution, 10 mL sodium carbonate solution and 10 mL sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated, the product was purified by preparative HPLC (chromatographic column: waters Xbridge 15025 mm 5 m; mobile phases: [A-HCl/H.sub.2O=0.04% v/v; B-ACN]B %: 5%-30%, 12 min]) to give the compound 48. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.78 (s, 1H) 7.63-7.73 (m, 1H) 7.43-7.51 (m, 1H) 7.28-7.35 (m, 1H) 6.83-6.99 (m, 2H) 6.52-6.66 (m, 2H) 5.62-5.82 (m, 1H) 5.14-5.30 (m, 1H) 4.90-5.07 (m, 2H) 4.73-4.81 (m, 2H) 3.44 (s, 2H) 3.09-3.12 (m, 4H) 2.34-2.46 (m, 4H) 2.26 (s, 3H) 1.46-1.68 (m, 8H) 1.31 (d, J=6.0 Hz, 6H) MS m/z: 569.3[M+H].sup.+

Embodiment 49: Synthesis of Compound 49

(342) ##STR00235##

(343) Synthetic Route:

(344) ##STR00236##

Step 1: Synthesis of Compound 49-1

(345) 2-bromo-6-acetonylpyridine (2 g, 10.10 mmol) was added into 20 mL DCM solution, then diethylaminosulfur trifluoride (4.85 g, 30.30 mmol) was added to the mixture at 0 C., then stirred for 5 min, and the temperature was raised to 35 C., stirred for 2 hours until the reaction was complete. The reaction mixture was slowly poured into 50 mL ice water, extracted by 50 mL EA, the organic phase was washed by 50 mL saturated brine, dried over excess anhydrous sodium sulfate, then filtered, evaporated, mixed, then purified by chromatography (EA/PE=1/15) to give 49-1. MS m/z: 361.1[M+H].sup.+

Step 2: Synthesis of Compound 49-2

(346) 49-1 (1.40 g, 6.31 mmol) and I1 (1.40 g, 6.31 mmol) were added into 1,4-dioxane solution, then N,N-dimethylethylenediamine (733.25 mg, 6.31 mmol, 904.13 L), potassium carbonate (1.13 g, 8.20 mmol) and cuprous iodide (1.20 g, 6.31 mmol) were added sequentially under nitrogen atmosphere, the temperature was raised to 95 C. and stirred at reflux for 12 hours until the reaction was complete. After cooled down to 25 C., the reaction was concentrated at 45 C. 50 mL water was added to quench the reaction, then extracted by 50 mL EA, the organic phase was washed by 50 mL saturated brine, dried over anhydrous sodium sulfate, then filtered, evaporated, mixed, purified by chromatography (EA/PE=1/10-1/4) to give 49-2. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.93 (s, 1H), 8.04-7.95 (m, 2H), 7.57 (d, J=7.4 Hz, 1H), 7.24 (s, 1H), 5.67-5.60 (m, 1H), 4.99 (d, J=10.0 Hz, 1H), 4.91-4.87 (m, 2H), 2.58 (s, 3H), 2.04-1.95 (m, 3H)

Step 3: Synthesis of Compound 49-3

(347) 49-2 (300.00 mg, 0.82 mmol) was added into DCM solution, then m-CPBA (203.52 mg, 1.00 mmol, 85% purity) was added, then stirred at 25 C. for 2 hours until the reaction was complete. The mixture was concentrated under reduced pressure at 45 C., and the filtrate was slowly added into 20 mL saturated sodium sulfite solution to quench the reaction, then extracted by 20 mL DCM, the organic phase was washed by 20 mL saturated brine, dried over anhydrous sodium sulfate, then filtered, evaporated to dry to give the crude product 49-3, which was used directly in the next step. MS m/z: 380.1[M+H].sup.+

Step 4: Synthesis of Compound 49

(348) Under nitrogen atmosphere, 49-3 (300 mg, 0.79 mmol) and 12 (205.12 mg, 0.79 mmol) was added into DCM solution, then DIPEA (306.59 mg, 2.37 mmol, 0.40 mL) was added, and stirred at 30 C. for 12 hours. The solvent therein was evaporated, then purified by preparative HPLC ((chromatographic column: waters Xbridge 15025 mm 5 m; mobile phases: [water (0.225% FA)-ACN]; B (ACN) %: 10%-35%, 12 min).)) to give 49. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.83 (s, 1H), 8.03 (d, J=8.4 Hz, 1H), 7.95-7.91 (m, 1H), 7.53 (d, J=7.6 Hz, 1H), 7.44 (br d, J=8.6 Hz, 2H), 6.92 (d, J=9.2 Hz, 2H), 5.67-5.63 (m, 1H), 4.94 (d, J=19.6 Hz, 1H), 4.84 (s, 3H) 3.17-3.14 (m, 4H), 2.49 (br s, 4H), 2.36 (s, 3H), 1.68-1.67 (m, 3H), 1.66-1.62 (m, 8H) MS m/z: 575.3[M+H].sup.+

Embodiment 50: Compound 50

(349) ##STR00237##

(350) Synthetic Route:

(351) ##STR00238##

Step 1: Synthesis of Compound 50-1

(352) 2-Bromo-6-fluoropyridine (3.00 g, 17.05 mmol) and cyclopropanol (1.09 g, 18.76 mmol) were dissolved in 30 mL THF, potassium tert-butoxide (2.47 g, 22.01 mmol) was added at 0 C., then stirred at 0 C. for 12 h. 50 mL water was added into, then extracted by 100 mL EA, the organic phase was washed by 100 mL brine, dried over anhydrous sodium sulfate, and the filtrate was concentrated to give 50-1. MS m/z: 216.0 [M+H].sup.+

Step 2: Synthesis of Compound 50-2

(353) I1 (2.00 g, 9.00 mmol) was added into 1,4-dioxane (30.00 mL), and 50-1 (2.12 g, 9.90 mmol), potassium carbonate (1.70 g, 12.33 mmol), cuprous iodide (1.71 g, 9.00 mmol) and N,N-dimethylethylenediamine (872.50 mg, 9.90 mmol, 1.08 mL) were added while stirring, the temperature was raised to 95 C. and stirred for 12 h under nitrogen atmosphere. 100 mL ammonia was added to the mixture, then extracted by 100 mL EA, the organic phase was washed by 100 mL brine, dried over anhydrous sodium sulfate, the filtrate was concentrated, crystallized in EtOAc to give 50-2. MS m/z: 356.1 [M+H].sup.+

Step 3: Synthesis of Compound 50

(354) Compound 50-2 (500.00 mg, 1.41 mmol) and m-CPBA (324.43 mg, 1.60 mmol) were added into 5 mL DCM, then reacted at 25 C. for 1 h. 12 (632.73 mg, 2.44 mmol) and DIPEA (501.13 mg, 3.88 mmol, 677.20 L) were added and stirred at 30 C. for 12 h. The reacted mixture was partitioned between 10 mL DCM and 10 mL, the organic phase was washed separately by 10 mL saturated sodium sulfite solution, 10 mL sodium carbonate solution and 10 mL sodium chloride solution, dried over anhydrous sulfuric acid, the filtrate was concentrated. The residue was purified by preparative HPLC (chromatographic column: waters Xbridge 15025 mm 5 m; mobile phases: [water (0.225% FA)-ACN]; B %: 10%-30%, 12 min) to give 50. .sup.1H NMR (400 MHz, CDCl.sub.3)

(355) 8.80 (s, 1H), 7.71 (t, J=8.0 Hz, 1H), 7.47-7.50 (m, 2H), 6.91 (d, J=8.8 Hz, 2H), 6.65 (d, J=8.4 Hz, 1H), 5.70-5.61 (m, 1H), 5.02-4.98 (m, 2H), 4.88 (d, J=6.4 Hz, 2H), 4.17-4.22 (m, 1H) 3.11-3.14 (m, 4H), 2.38 (br, 4H), 2.28 (s, 3H), 1.59-1.65 (m, 8H), 0.78 (d, J=6.4 Hz, 4H), MS m/z: 569.3[M+H].sup.+

Embodiment 51: Compound 51

(356) ##STR00239##

(357) Synthetic Route:

(358) ##STR00240##

Step 1: Synthesis of Compound 51-1

(359) Anhydrous ethanol (2.92 g, 63.33 mmol, 3.70 mL) was added into THF (20.00 mL) solution, then sodium hydride (1.52 g, 63.33 mmol) was added to the mixture and stirred for 5 min, then 2,6-dibromopyridine (5.00 g, 21.11 mmol) was added, the mixture was stirred at 25 C. for 2 hours until the reaction was complete. The reaction mixture was slowly poured into 50 mL ice water, then extracted by 50 mL EA, the organic phase was washed by 50 mL saturated brine, dried over anhydrous sodium sulfate, then filtered, evaporated, mixed and purified by chromatography (EA/PE=1/20-1/10) to give 51-1. .sup.1H NMR (400 MHz, CDCl.sub.3) 7.37-7.33 (m, 1H), 6.98 (d, J=7.2 Hz, 1H), 6.61 (d, J=8.0 Hz, 1H), 4.33-4.28 (m, 2H), 1.35-1.33 (m, 3H)

Step 2: Synthesis of Compound 51-2

(360) Compound 51-1 (2.00 g, 9.90 mmol) and Intermediate 11 (2.20 g, 9.90 mmol) were added into 1,4-dioxane solution, then N,N-diethylethylenediamine (1.27 g, 10.89 mmol), potassium carbonate (1.37 g, 9.90 mmol) and cuprous iodide (1.89 g, 9.90 mmol) were added sequentially under nitrogen atmosphere, the temperature was raised to 95 C. and the reaction was stirred at reflux for 12 hours until it was complete. Then the mixture was cooled down to r.t. and concentrated, quenched by 50 mL water, then extracted by 50 mL EA, the organic phase was washed by 50 mL saturated brine, dried over anhydrous sodium sulfate, then filtered, evaporated and purified by chromatography (EA/PE=1/10-1/4) to give 51-2. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.90 (s, 1H), 7.75-7.71 (m, 1H), 7.35 (d, J=8.0 Hz, 1H), 6.67 (d, J=8.2 Hz, 1H), 5.71-5.64 (m, 1H), 5.04 (dd, J=10.0 Hz, 1H), 5.05-4.80 (m, 1H), 4.81 (d, J=6.4 Hz, 1H), 4.34-4.29 (m, 2H), 2.55 (s, 3H), 1.56 (d, J=2.4 Hz, 1H), 1.42-1.38 (m, 3H)

Step 3: Synthesis of Compound 51-3

(361) Compound 51-2 (300.00 mg, 0.87 mmol) was added into DCM solution, m-CPBA (215.37 mg, 1.06 mmol, 85% purity) was added into the reaction mixture, then stirred at 25 C. for 2 hours, until the reaction was complete. The reaction mixture was concentrated, then slowly added into 20 mL saturated sodium sulfite solution to quench the reaction, then extracted by 20 mL DCM, the organic phase was washed by 20 mL saturated brine, dried over anhydrous sodium sulfate, then filtered and evaporated to give the crude product 51-3 which was used directly in the next step.

Step 4: Synthesis of Compound 51

(362) Crude product 51-3 and Intermediate 12 (216.52 mg, 0.83 mmol) was added into DCM solution, then DIPEA (323.64 mg, 2.50 mmol, 0.44 mL) was added, under N.sub.2 atmosphere, the mixture was stirred at 30 C. for 12 hours until the reaction was complete. The solvent was evaporated, then purified by preparative HPLC (chromatographic column: waters Xbridge 15025 mm 5 m; mobile phases: [water (0.225% FA)-ACN]; B (acetonitrile) %: 10%-20%, 12 min) to give 51. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.76 (br s, 1H), 7.71-7.67 (m, 1H), 7.47 (br d, J=8.0 Hz, 2H), 7.34 (br d, J=7.6 Hz, 1H), 6.87 (br d, J=8.4 Hz, 2H), 6.64 (br d, J=8.2 Hz, 1H), 5.71-5.63 (m, 1H), 5.03-4.92 (m, 2H), 4.75 (br d, J=5.6 Hz, 2H), 4.32 (q, J=7.2 Hz, 2H), 3.11-2.88 (m, 8H), 2.71 (s, 3H), 1.84 (br s, 4H), 1.70 (br s, 4H), 1.38-1.36 (m, 3H) MS m/z: 555.3[M+H].sup.+

Embodiment 52: Compound 52

(363) ##STR00241##

(364) Synthetic Route:

(365) ##STR00242##

Step 1: Synthesis of Compound 52-1

(366) 2-Bromo-6-fluoro-pyridine (2 g, 11.36 mmol), (R)-3-hydroxytetrahydrofuran (1.10 g, 12.50 mmol, 1.00 mL) and THF (20 mL) were added into a pre-dried 100 mL three neck flask, then t-BuOK (1.65 g, 14.65 mmol) was added at 0 C., replaced with nitrogen for 3 times, the reaction was kept for 0.5 hours at 25 C. under nitrogen atmosphere. 20 mL Water was added, extracted by 30 mL3 EA, the organic phase was dried over anhydrous sodium sulfate, filtered and evaporated to give 52-1. .sup.1H NMR (400 MHz, CDCl.sub.3) =7.50-7.34 (m, 1H), 7.06 (d, J=7.4 Hz, 1H), 6.69 (d, J=8.2 Hz, 1H), 5.57 (tdd, J=2.0, 4.4, 6.4 Hz, 1H), 4.05-3.83 (m, 4H), 2.35-2.19 (m, 1H), 2.19-2.04 (m, 1H)

Step 2: Synthesis of Compound 52-2

(367) Compound I1 (1 g, 4.50 mmol) and 1,4-dioxane (20 mL) were added into a pre-dried 100 mL three neck flask, followed by adding compound 52-1 (1.21 g, 4.95 mmol), cuprous iodide (856.85 mg, 4.50 mmol), potassium carbonate (851.89 mg, 6.16 mmol), and N,N-dimethylethylenediamine (436.26 mg, 4.95 mmol, 540.60 L), replaced with nitrogen for 3 times, the reaction was kept for 16 hr at 95 C. under nitrogen atmosphere. 30 mL ammonia was added, then extracted by 30 mL3 EA, the organic phase was dried over anhydrous sodium sulfate, filtered and evaporated. The crude product was purified by chromatography (SiO.sub.2, 100-200 mesh, PE/EtOAc=10/1-1/1) to give the 52-2. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.93 (s, 1H), 7.78 (t, J=8.0 Hz, 1H), 7.39 (d, J=7.7 Hz, 1H), 6.73 (d, J=8.2 Hz, 1H), 5.69 (tdd, J=6.2, 10.4, 16.9 Hz, 1H), 5.51 (dt, J=2.3, 4.3 Hz, 1H), 5.08 (d, J=10.2 Hz, 1H), 4.98 (dd, J=1.0, 17.1 Hz, 1H), 4.77 (t, J=6.5 Hz, 2H), 4.05-3.98 (m, 2H), 3.95-3.86 (m, 2H), 2.57 (s, 3H), 2.32-2.20 (m, 1H), 2.17-2.05 (m, 1H)

Step 3: Synthesis of Compound 52

(368) Compound 52-2 (0.2 g, 518.89 mol) and DCM (2 mL) were added into a pre-dried 40 mL flask, followed by adding m-CPBA (136.95 mg, 674.55 mol, 85% purity), replaced with nitrogen for 3 times, and the reaction was kept at 25 C. for 1 hr under nitrogen atmosphere. DCM (1 mL), DIEA (193.17 mg, 1.49 mmol, 260.34 L) and 12 (135.69 mg, 523.12 mol) were added, and replaced with nitrogen, the reaction was kept at 25 C. for 16 hr under nitrogen atmosphere. 5 mL saturated sodium sulfite solution was added, then extracted with 10 mL2 DCM, dried over anhydrous sodium sulfate, the organic phase was filtered and evaporated to dry. The residue was purified by prep-HPLC (chromatographic column: Agela Durashell C18 15025 mm 5 m; mobile phases: [water (10 mM NH.sub.4HCO.sub.3)-ACN]; B %: 35%-55%, 10.5 min), freeze-dried to give 52. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.82 (s, 1H), 7.75 (t, J=7.9 Hz, 1H), 7.43 (br dd, J=7.8, 19.1 Hz, 3H), 6.92 (d, J=8.9 Hz, 2H), 6.69 (d, J=8.2 Hz, 1H), 5.76-5.64 (m, 1H), 5.54 (br s, 1H), 5.06 (d, J=9.8 Hz, 1H), 4.98 (d, J=16.8 Hz, 1H), 4.88-4.65 (m, 4H), 4.06-3.87 (m, 4H), 3.18-3.11 (m, 4H), 2.41 (br s, 3H), 2.31 (s, 3H), 2.29-2.22 (m, 1H), 2.17-2.09 (m, 1H), 1.63-1.58 (m, 8H) MS m/z: 597.4.sup.+

Embodiment 53: Compound 53

(369) ##STR00243##

(370) Synthetic Route:

(371) ##STR00244##

Step 1: Synthesis of Compound 53-1

(372) 2-Bromo-6-fluoropyridine (2 g, 11.36 mmol) and 3-hydroxyoxetane (926.05 mg, 12.50 mmol) were dissolved in anhydrous THF (10 mL), potassium tert-butoxide (1.65 g, 14.66 mmol) was added at 0 C., and stirred at 0 C. for 2 h. The reaction mixture was added into water (20 mL), then extracted with EA (220 mL), the organic phase was washed by saturated brine (30 mL), dried over anhydrous sodium sulfate, and the filtrate was concentrated to give 53-1. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 7.40-7.40 (m, 1H) 7.36-7.65 (m, 1H) 7.11 (d, J=7.6 Hz, 1H) 5.60-5.71 (m, 1H) 4.73-5.01 (m, 2H) 4.60-4.79 (m, 2H) MS m/z: 230.06 [M+H].sup.+

Step 2: Synthesis of Compound 53-2

(373) I1 (805.11 mg, 3.62 mmol) and 53-1 (1 g, 4.35 mmol) were added into 1,4-dioxane (20.00 mL), followed by cuprous iodide (689.86 mg, 3.62 mmol), potassium carbonate (685.85 mg, 4.96 mmol) and N,N-methylethylenediamine (351.24 mg, 3.98 mmol, 428.86 L), the reaction mixture was heated to 95 C. under nitrogen atmosphere, then reacted for 12 h. After adding ammonia (100 L), the reacted mixture was then extracted by EA (250 L), the organic phase was washed by saturated brine (50 mL), dried over anhydrous sodium sulfate, the filtrate was concentrated to give the crude product, which was purified by silica gel column chromatography (silica gel mesh: 100-200 mesh; PE/EA=20/1-0/1) to give 53-2. MS m/z: 371.41 [M+H].sup.+

Step 3: Synthesis of Compound 53

(374) 53-2 (200 mg, 538.48 mol) was added into anhydrous DCM (6 mL), followed by m-CPBA (159.14 mg, 783.85 mol, 85% purity), the mixture was stirred at 25 C. for 1.5 hours. Then DIPEA (173.47 mg, 1.34 mmol, 233.79 L), 12 (167.12 mg, 644.27 mol) were added, the reaction mixture was stirred at 35 C. for 16 hours. The reaction mixture was evaporated to dry, then saturated sodium sulfite solution (10 mL) was added, and extracted by DCM (310 mL), the organic phase was dried over anhydrous sodium sulfate, then filtered, concentrated under reduced pressure to give a residue, which was purified by prep-HPLC (chromatographic column: Agela Durashell C18 15025 mm 5 m; mobile phases/[water (10 mM NH.sub.4HCO.sub.3)-ACN]; B (acetonitrile) %: 28%-58%, 10.5 min) and thin layer chromatography silica gel plate (DCM/MeOH=10/1) to give the 53. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 8.80 (s, 1H) 7.76 (br t, J=8.4 Hz, 1H) 7.37-7.47 (m, 3H) 6.91 (br d, J=8.0 Hz, 2H) 6.72 (br d, J=8.0 Hz, 1H) 5.48-5.78 (m, 2H) 4.90-5.04 (m, 1H) 4.90-5.07 (m, 3H) 4.73 (br t, J=6.4 Hz, 2H) 4.64 (br d, J=6.8 Hz, 2H) 3.13 (br d, J=4.85 Hz, 4H) 2.45 (br s, 4H) 2.32 (s, 3H) 1.62-1.65 (m, 8H) MS m/z: 582.69 [M+H].sup.+

Embodiment 54: Compound 54

(375) ##STR00245##

(376) Synthetic Route:

(377) ##STR00246##

Step 1: Synthesis of Compound 54-1

(378) 2-Bromo-6-fluoropyridine (2 g, 11.36 mmol) and cyclobutanol (819.12 mg, 11.36 mmol) were dissolved in anhydrous THF (10 mL), potassium tert-butoxide (1.65 g, 14.65 mmol) was added at 0 C. and then stirred at 0 C. for 2 h. Water (20 mL) was added into the reaction mixture, and extracted by EA (220 mL), the organic phase was washed by saturated brine (30 mL), dried over anhydrous sodium sulfate, the filtrate was concentrated to give 54-1. .sup.1H NMR (400 MHz, CDCl.sub.3) =7.46-7.30 (m, 1H), 7.01 (d, J=7.5 Hz, 1H), 6.60 (d, J=8.2 Hz, 1H), 5.14 (quin, J=7.5 Hz, 1H), 2.49-2.38 (m, 2H), 2.17-2.02 (m, 2H), 1.72-1.47 (m, 2H) MS m z: 228.09 [M+H].sup.+

Step 2: Synthesis of Compound 54-2

(379) I1 (812.07 mg, 3.65 mmol, 1 eq) and 54-1 (1 g, 4.38 mmol) were added into 1,4-dioxane (20.00 mL), followed by cuprous iodide (695.83 mg, 3.65 mmol), potassium carbonate (691.80 mg, 5.01 mmol) and N,N-dimethylethylenediamine (354.27 mg, 4.02 mmol, 432.57 L), the temperature was raised to 95 C. under nitrogen atmosphere, then the reaction was kept for 12 h. Ammonia (100 mL) was added into the reacted mixture, then extracted by EA (250 mL), the organic phase was washed by saturated brine (50 mL), dried over anhydrous sodium sulfate, the filtrate was concentrated to give the crude product, which was purified by silica gel column chromatography (silica gel mesh: 100-200 mesh; PE/EA=20/1-10/1) to give 54-2. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.85 (s, 1H), 7.67 (t, J=7.9 Hz, 1H), 7.29 (d, J=7.7 Hz, 1H), 6.59 (d, J=8.2 Hz, 1H), 5.73-5.48 (m, 1H), 5.19-4.93 (m, 2H), 4.88 (br d, J=17.2 Hz, 1H), 4.75 (d, J=6.2 Hz, 2H), 2.50 (s, 3H), 2.40-2.23 (m, 2H), 2.07-2.14 (m, 2H), 1.58-1.82 (m, 2H) MS m z: 369.44 [M+H].sup.+

Step 3: Synthesis of Compound 54-3

(380) 54-2 (200 mg, 541.36 mol) was added into anhydrous DCM (5 mL), followed by m-CPBA (159.99 mg, 788.04 mol, 85% purity), the reaction mixture was stirred at 25 C. for 1.5 hours. DIPEA (174.36 mg, 1.35 mmol, 234.99 L) and I2 (153.98 mg, 593.61 mol) were added, the reaction mixture was stirred at 35 C. for 16 hours. Saturated sodium sulfite solution (10 mL) was added, then extracted by DCM (310 mL), the organic phase was dried over anhydrous sodium sulfate, then filtered, concentrated under reduced pressure to give a residue, which was purified by prep-HPLC (chromatographic column: Agela Durashell C18 15025 mm 5 m; mobile phases: [water (10 mM NH.sub.4HCO.sub.3)-ACN]; B (acetonitrile) %: 46%-66%, 10.5 min) to give the 54. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.83 (s, 1H), 7.74 (t, J=7.9 Hz, 1H), 7.48 (br d, J=8.3 Hz, 2H), 7.40 (br d, J=7.8 Hz, 1H), 6.94 (d, J=9.0 Hz, 2H), 6.64 (d, J=8.2 Hz, 1H), 5.75-5.66 (m, 1H), 4.95-5.16 (m, 3H), 4.79 (br d, J=5.8 Hz, 2H), 3.18-3.13 (m, 4H), 2.52-2.41 (m, 6H), 2.33 (s, 3H), 2.18 (ddd, J=2.7, 7.6, 9.8 Hz, 2H), 1.92-1.82 (m, 1H), 1.76-1.63 (m, 11H) MS m/z: 580.72 [M+H].sup.+

Embodiment 55: Compound 55

(381) ##STR00247##

(382) Synthetic Route:

(383) ##STR00248##

Step 1: Synthesis of Compound 55-1

(384) 2-Bromo-6-fluoropyridine (9.99 g, 56.75 mmol) and S-3-hydroxytetrahydrofuran (5 g, 56.75 mmol) were dissolved in THF (100 mL), potassium tert-butoxide (8.28 g, 73.78 mmol) was added at 0 C., then reacted at 25 C. for 12 h. The reaction mixture was extracted by 50 mL water and 100 mL EA, the organic phase was washed by 100 mL brine, dried over anhydrous sodium sulfate, filtered and concentrated to give 55-1, which was used directly in the next step.

Step 2: Synthesis of Compound 55-2

(385) 55-1 (576.54 mg, 2.36 mmol), I1 (500 mg, 2.25 mmol), cuprous iodide (428.43 mg, 2.25 mmol), potassium carbonate (432.17 mg, 3.13 mmol) and N,N-methylethylenediamine (218.13 mg, 2.47 mmol, 266.33 L, 1.10 eq) was added into 1,4-dioxane (15 mL), replaced with nitrogen 3 times and, the mixture was stirred at 95 C. for 13 hr under nitrogen atmosphere. 10 mL ammonia was added to quench the reaction, then extracted with 30 mL DCM, the organic phase was washed by 20 mL saturated brine, dried over anhydrous sodium sulfate, then filtered and evaporated to dry to give 55-2, which was used directly in the next step. MS m/z: 386.3 [M+H].sup.+

Step 3: Synthesis of Compound 55

(386) 55-2 (200 mg, 518.89 mol) was dissolved in DCM (2 mL), followed by adding m-CPBA (144.32 mg, 710.88 mol, 85% purity), the mixture was stirred at 25 C. for 1 h. DIPEA (167.66 mg, 1.30 mmol, 225.95 L) and 12 (134.59 mg, 518.88 mol) were added, then reacted at 25 C. for 12 h. Saturated sodium sulfite solution 5 mL was added into the reaction mixture, then extracted by DCM 10 mL, the organic phase was dried over anhydrous sodium sulfate, then filtered and evaporated to dry to give the crude product. The crude product was purified by prep-HPLC (chromatographic column: waters Xbridge 15025 mm 5 m; mobile phases: [A-HCl/H.sub.2O=0.04% v/v; B-ACN] B %: 1%-25%, 12 min]) and adjusted pH=7 with basic resin, then filtered and concentrated to give 55.

(387) .sup.1H NMR (400 MHz, CDCl.sub.3) =8.80 (s, 1H), 7.73 (t, J=8.0 Hz, 1H), 7.38-7.44 (m, 3H), 6.90 (d, J=8.8 Hz, 2H), 6.67 (d, J=8.0 Hz, 1H), 5.74-5.62 (m, 1H), 5.50-5.52 (m, 1H), 5.07-4.93 (m, 2H), 4.71 (br t, J=6.1 Hz, 2H), 4.05-3.84 (m, 4H), 3.17-3.08 (m, 4H), 2.38 (br s, 4H), 2.28 (s, 3H), 2.27-2.20 (m, 1H), 2.15-2.06 (m, 1H), 1.68-1.58 (m, 10H), MS m/z: 597.3 [M+H].sup.+

Embodiment 56: Compound 56

(388) ##STR00249##

(389) Synthetic Route:

(390) ##STR00250##

Step 1: Synthesis of Compound 56-1

(391) Sodium hydrogen (909.07 mg, 22.73 mmol, 60% purity) was dissolved in anhydrous THF (5 mL), the reaction was placed in an ice bath, replaced with nitrogen for 3 times, 2-bromo-6-fluoropyridine (2 g, 11.36 mmol), and 1-methylimidazolidinone (2.28 g, 22.73 mmol) were added at 0 C., the mixture was stirred at 70 C. for 16 hours. Water (20 mL) was added to quench the reaction, then extracted with EA (320 mL), the organic phase was washed by saturated brine (20 mL), dried over anhydrous sodium sulfate, the filtrate was concentrated to give the crude product, the crude product was purified by column to give the 56-1. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.23 (d, J=8.5 Hz, 1H), 7.43 (t, J=7.7 Hz, 1H), 7.05 (d, J=7.5 Hz, 1H), 4.10-3.93 (m, 2H), 3.53-3.40 (m, 2H), 2.95-2.85 (m, 3H), MS m/z: 256.10 [M+H].sup.+

Step 2: Synthesis of Compound 56-2

(392) I1 (867.89 mg, 3.90 mmol), 56-1 (1 g, 3.90 mmol), cuprous iodide (743.66 mg, 3.90 mmol), potassium carbonate (755.54 mg, 5.47 mmol) and N,N-dimethylethylenediamine (378.62 mg, 4.30 mmol, 469.17 L) were added into 1,4-dioxane (20.00 mL), the temperature was raised to 95 C. under nitrogen atmosphere, then reacted for 16 h. Ammonia (50 mL) was added into the reacted mixture, extracted by EA (350 mL), the organic phase was washed by saturated brine (50 mL), dried over anhydrous sodium sulfate, the filtrate was concentrated to give the crude product, the residue was separated by an automatic column machine (PE/EA=10/1-5/1) to give the 56-2. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 8.94 (s, 1H), 8.29 (d, J=8.4 Hz, 1H) 7.83 (t, J=8.41 Hz, 1H) 7.43 (d, J=7.6 Hz, 1H) 5.68-5.75 (m, 1H), 4.95-5.09 (m, 2H), 4.82 (d, J=6.0 Hz, 2H) 4.03 (t, J=8.4 Hz, 2H) 3.53 (t, J=8.4 Hz, 2H) 2.96 (s, 3H) 2.59 (s, 3H) MS m/z: 397.45 [M+H].sup.+

Step 3: Synthesis of Compound 56

(393) 56-2 (200 mg, 503.20 mol) was added into anhydrous DCM (10 mL), followed by adding m-CPBA (148.71 mg, 689.39 mol, 85% purity), the reaction mixture was stirred at 25 C. for 1.5 hours. DIPEA (162.55 mg, 1.26 mmol, 219.06 L) and 12 (156.59 mg, 603.70 mol) were added, the reaction mixture was stirred at 35 C. for 16 hours. Saturated sodium sulfite solution (10 mL) was added to quench the reaction, then extracted by DCM (310 mL), the organic phase was dried over anhydrous sodium sulfate, then filtered, concentrated under reduced pressure to give a residue, the residue was purified by prep-HPLC (chromatographic column/Xtimate C18 15025 mm5 m; mobile phases/[water (10 mM NH.sub.4HCO.sub.3)-ACN]; B (acetonitrile) %: 30%-50%, 10.5 min) to give 56. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.79 (s, 1H), 8.20 (d, J=8.4 Hz, 1H), 7.77 (t, J=8.0 Hz, 1H), 7.47-7.30 (m, 3H), 6.91-6.89 (d, J=8.8 Hz, 2H), 5.73-5.61 (m, 1H), 4.94-5.04 (m, J=17.2 Hz, 2H), 4.73 (br d, J=6.2 Hz, 2H), 4.01 (t, J=7.6 Hz, 2H) 3.48 (t, J=7.6 Hz, 2H) 3.14-3.10 (m, 4H), 2.92 (s, 3H), 2.41 (br s, 4H), 2.30 (s, 3H), 1.66-1.61 (m, 8H) MS m/z: 608.73 [M+H].sup.+

Embodiment 57: Compound 57

(394) ##STR00251##

(395) Synthetic Route:

(396) ##STR00252##

Step 1: Synthesis of Compound 57-1

(397) 2-Bromo-6-fluoropyridine (2 g, 11.36 mmol), potassium carbonate (3.14 g, 22.73 mmol) and oxazolidinone (1.98 g, 22.73 mmol) was placed in DMF (5 mL), and stirred at 90 C. for 16 hours. Water (10 mL) was added into the reaction mixture, then extracted by EA (10 mL), washed by water (25 mL), the organic phase was washed by brine (5 mL), dried over anhydrous sodium sulfate, the filtrate was concentrated to give the crude product, the crude product was purified by an automatic column machine COMBI-FLASH (PE/EA=50/1-2/1) to give 57-1. MS m/z: 243.06 [M+H].sup.+

Step 2: Synthesis of Compound 57-2

(398) I1 (182.89 mg, 822.85 mol) and 57-1 (200 mg, 822.85 mol) were added into 1,4-dioxane (5.00 mL), followed by adding cuprous iodide (156.71 mg, 822.85 mol), potassium carbonate (159.22 mg, 1.15 mmol) and N,N-dimethylethylenediamine (79.79 mg, 905.14 mol, 98.87 L), the temperature was raised to 95 C. under nitrogen atmosphere, then reacted for 16 h. Ammonia (20 mL) was added into the reacted mixture, then extracted by EA (310 mL), the organic phase was washed by saturated brine (20 mL), dried over anhydrous sodium sulfate, the filtrate was concentrated to give the crude product, which was separated by an automatic column machine COMBI-FLASH (PE/EA=10/1-1/1) to give 57-2. MS m/z: 384.41 [M+H].sup.+

Step 3: Synthesis of Compound 57

(399) 57-2 (195 mg, 507.27 mol) was added into anhydrous DCM (10 mL), followed by adding m-CPBA (149.91 mg, 738.39 mol, purity is 85%), the reaction mixture was stirred at 25 C. for 1.5 hours. DIPEA (163.80 mg, 1.27 mmol, 220.76 L), 12 (157.81 mg, 608.37 mol) was then added, the reaction mixture was stirred at 35 C. for 16 hours. Saturated sodium sulfite solution (10 mL) was added into the reaction mixture, then extracted by DCM (310 mL), the organic phase was dried over anhydrous sodium sulfate, then filtered, concentrated under reduced pressure to give a residue, which was purified by prep-HPLC (chromatographic column: Xtimate C18 15025 mm5 m; mobile phases: [water (10 mM NH.sub.4HCO.sub.3)-ACN]; B (acetonitrile) %: 25%-55%, 10.5 min). then thin layer chromatography silica gel plate (DCM/MeOH=10/1) to give the 57. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.79 (s, 1H), 8.12 (d, J=7.9 Hz, 1H), 7.84 (t, J=8.1 Hz, 1H), 7.55 (d, J=7.9 Hz, 1H), 7.40 (br d, J=8.8 Hz, 2H), 6.89 (d, J=9.0 Hz, 2H), 5.66 (tdd, J=6.3, 10.3, 16.9 Hz, 1H), 5.03 (dd, J=1.0, 10.2 Hz, 1H), 4.94 (dd, J=1.1, 17.1 Hz, 1H), 4.69 (br d, J=6.3 Hz, 2H), 4.49 (t, J=8.1 Hz, 2H), 4.23 (t, J=8.0 Hz, 2H), 3.17-3.08 (m, 4H), 2.39 (br s, 4H), 2.29 (s, 3H), 1.64-1.56 (m, 8H) MS m/z: 595.69 [M+H].sup.+

Embodiment 58: Compound 58

(400) ##STR00253##

Step 1: Synthesis of Compound 58-1

(401) 6-Bromopyridin-2-amine (2 g, 11.56 mmol) was dissolved in anhydrous THF (5 mL), the reaction was placed in an ice bath, and replaced with nitrogen for 3 times, 1-chloro-2-isocyanato-ethane (1.83 g, 17.34 mmol, 1.48 mL) was added at 0 C., the reaction system was stirred at 25 C. for 16 hours. Water (10 mL) was added into the reaction mixture, then extracted with EA (310 mL), the organic phase was washed by saturated brine (210 mL), dried over anhydrous sodium sulfate, the filtrate was concentrated to give the crude product, 20 mL PE was added into the crude product and stirred for 1 hour, then filtered, the filter cake was washed by PE (220 mL) and collected, evaporated under increased pressure to give a white solid compound (1.34 g, 4.81 mmol, yield 41.62%). MS m/z: 278.53 [M+H].sup.+

Step 2: Synthesis of Compound 58-2

(402) Sodium hydride (384.84 mg, 9.62 mmol, 60% purity) was added into anhydrous THF (20 mL), the reaction system was placed in an ice bath, replaced with nitrogen for 3 times, 58-1 (1.34 g, 4.81 mmol,) was added at 0 C., the mixture was stirred to react at 25 C. for 15 min, then placed in an oil bath at 70 C. and stirred to react for 2 hours, water (10 mL) was added into the reaction mixture, and extracted by EA (220 mL), the organic phase was washed by saturated brine (10 mL), dried over anhydrous sodium sulfate, the filtrate was concentrated to give 58-2. MS m/z: 242.07 [M+H].sup.+

Step 3: Synthesis of Compound 58-3

(403) 58-2 (1.1 g, 4.54 mmol), di-tert-butyl dicarbonate (991.75 mg, 4.54 mmol, 1.04 mL) and triethylamine (1.38 g, 13.63 mmol, 1.90 mL) were dissolved in anhydrous THF (5 mL), and replaced with nitrogen for 3 times, then DMAP (55.52 mg, 454.41 mol) was added, the reaction system was stirred to react at 60 C. 10 hours. The reaction mixture was concentrated under reduced pressure and evaporated to dry, saturated brine 10 mL was added, then extracted by EA 20 mL, the organic phase was washed by saturated brine 10 mL, dried over anhydrous sodium sulfate, then filtered and evaporated under reduced pressure to give the crude product. The crude product was purified by an automatic column machine COMBI-FLASH (PE/EA=20/1-5/1) to give 58-3. MS m/z: 342.19 [M+H]+

Step 4: Synthesis of Compound 58-4

(404) I1 (194.86 mg, 876.71 mol), 58-3 (300 mg, 876.71 mol), cuprous iodide (166.97 mg, 876.71 molq), potassium carbonate (169.64 mg, 1.23 mmol) and N,N-dimethylethylenediamine (85.01 mg, 964.38 mol, 105.34 L) were added into 1,4-dioxane (5 mL), the temperature was raised to 95 C. under nitrogen atmosphere, the reaction was kept for 16 h. Ammonia (10 mL) was added into the reacted mixture, then extracted by EA (310 mL), the organic phase was washed by saturated brine (10 mL), dried over anhydrous sodium sulfate, the filtrate was concentrated to give the crude product, which was separated and purified by an automatic column machine COMBI-FLASH (PE/EA=10/1-5/1) to give the 58-4. MS m/z: 483.54 [M+H].sup.+

Step 5: Synthesis of Compound 58-5

(405) 58-4 (330 mg, 682.46 mol) was added into anhydrous DCM (10 mL), followed by adding m-CPBA (161.35 mg, 794.74 mol, 85% purity), the reaction mixture was stirred at 25 C. for 1.5 hours. DIPEA (219.91 mg, 1.70 mmol, 296.37 L) and 12 (211.86 mg, 816.75 mol) were added, the reaction mixture was stirred at 35 C. for 16 hours. Saturated sodium sulfite solution (10 mL) was added into the reaction mixture, then extracted by DCM (310 mL), the organic phase was dried over anhydrous sodium sulfate, then filtered and concentrated under reduced pressure to give the crude product 58-5. MS m/z: 694.82 [M+H].sup.+

Step 6: Synthesis of Compound 58

(406) DCM (12 mL) and trifluoroacetate (3 mL) were added into 58-5 (529 mg, crude product), then replaced with nitrogen for 3 times, the reaction system was stirred to react at 25 C. for 2 hours. The reaction system was directly concentrated under reduced pressure and evaporated to give a residue, the residue was purified by prep-HPLC (separated chromatographic column/Nano-micro Kromasil C18 10030 mm 5 m; mobile phases/[water (0.1% TFA)-ACN]; B (acetonitrile) %/5%-25%, 10 min), then by thin layer chromatography silica gel plate (DCM/MeOH=10/1) to give the 58. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.83 (s, 1H), 8.22 (d, J=8.3 Hz, 1H), 7.81 (t, J=8.0 Hz, 1H), 7.55-7.30 (m, 3H), 6.93 (d, J=9.0 Hz, 2H), 5.76-5.66 (m, 1H), 4.98-5.07 (m, 2H), 4.84 (s, 1H), 4.77 (br d, J=6.0 Hz, 2H), 4.18 (t, J=8.0 Hz, 2H), 3.61 (t, J=8.0 Hz, 2H), 3.20-3.13 (m, 4H), 2.65 (br s, 4H), 2.47 (s, 3H), 1.71 (br s, 8H) MS m/z: 594.71 [M+H].sup.+

Embodiment 59: Compound 59

(407) ##STR00254##

(408) Synthetic Route:

(409) ##STR00255##

Step 1: Synthesis of Compound 59-1

(410) In a pre-dried 100 mL thumb flask, 2,6-dibromopyridine (1 g, 4.22 mmol, 255.10 L), dimethylphosphine oxide (329.47 mg, 4.22 mmol), Pd (PPh.sub.3).sub.4 (211.88 mg, 183.36 mol), acetonitrile (10 mL) and TEA (1.54 g, 15.22 mmol, 2.12 mL) were added. The reaction mixture was replaced by nitrogen for 3 times, then placed in an oil bath at 90 C. and stirred for 13 hours. The reaction mixture was concentrated. The residue was purified by a silica gel column (100-200 mesh silica gel, DCM:MeOH=I/0-5/1) to give the crude product 59-1. .sup.1H NMR (400 MHz, CDCl.sub.3): 8.10-8.07 (m, 1H), 7.73-7.68 (m, 1H), 7.59-7.57 (m, 1H), 1.80-1.76 (d, J=13.6 Hz, 6H) MS m/z: 234.1 [M+H].sup.+

Step 2: Synthesis of Compound 59-2

(411) In a pre-dried thumb flask, 59-1 (0.8 g, 3.42 mmol), I1 (690.72 mg, 3.11 mmol), cuprous iodide (591.84 mg, 3.11 mmol), potassium carbonate (588.42 mg, 4.26 mmol) N,N-dimethylethylenediamine (301.33 mg, 3.42 mmol, 367.92 L) and 1,4-dioxane (20 mL) were added in sequence, the reaction mixture was replaced by nitrogen for 3 times, then heated and stirred in an oil bath at 95 C. for 13 hours. For another batch, 59-1 (200.85 mg, 858.21 mol), I1 (190.75 mg, 858.21 mol), cuprous iodide (163.45 mg, 858.21 mol), potassium carbonate (162.50 mg, 1.18 mmol), N,N-dimethylethylenediamine (83.22 mg, 944.04 mol, 101.61 L) and 1,4-dioxane (5 mL) were added sequentially in a pre-dried thumb flask, the reaction mixture was replaced by nitrogen for 3 times, then heated and stirred in an oil bath at 95 C. for 13 hours. Half of the reaction mixture and previous batch of the reaction mixture were combined and directly evaporated. The residue was purified by a silica gel column (100-200 mesh silica gel, PE:EA=5/1-0/1) to give 0.7 g pale brown oily product. 0.5 g of the oily product was added into 20 mL water, then extracted by 60 mL DCM for 3 times, the organic phase was dried, then filtered and evaporated to dry. The residue was purified by a silica gel column (100-200 mesh silica gel, PE:EA=5/1-0/1) to give 59-2. .sup.1H NMR (400 MHz, CDCl.sub.3): 8.97 (s, 1H), 8.09-8.06 (m, 1H), 7.70-7.43 (m, 2H), 5.71-5.64 (m, 1H), 5.07-5.05 (d, J=10.4 Hz, 1H), 4.93-4.88 (m, 1H), 4.81-4.79 (d, J=6.4 Hz, 2H), 2.61 (s, 3H), 1.81-1.75 (m, 6H)

Step 3: Synthesis of Compound 59

(412) (50 mg, 133.20 mol) was dissolved in dichloromethane (2 mL), then m-CPBA (41.99 mg, 206.82 mol, 85% purity) was added and stirred at 25 C. for 1 h. 12 (34.55 mg, 133.19 mol) and DIPEA (47.34 mg, 366.27 mol, 63.80 L) were added and stirred at 35 C. for 2 h. The reacted mixture was partitioned between dichloromethane (10 mL) and water (10 mL), the organic phase was washed separately by Na.sub.2SO.sub.3 solution 10 mL, Na.sub.2CO.sub.3 solution 10 mL and NaCl solution 10 mL, dried over anhydrous sodium sulfate, then filtered, the filtrate was concentrated to give a residue. The residue was purified by prep-HPLC (chromatographic column: waters Xbridge Prep OBD C18 15030 mm 5 m; mobile phases: [A-HCl/H.sub.2O=0.04% v/v; B-ACN] B %: 1%-25%, 12 min]) to give the 59.

(413) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 8.85 (s, 1H) 8.00-8.17 (m, 3H) 7.44 (d, J=7.6 Hz, 2H) 6.96 (d, J=7.2 Hz, 2H) 5.66-5.83 (m, 1H) 4.88-5.12 (m, 2H) 4.76 (d, J=6.0 Hz, 2H) 3.18 (br, 4H) 2.42 (br, 4H) 2.32 (s, 3H) 1.80 (dd, J=2.0, 13.8 Hz, 6H), 1.61-1.63 (m, 8H) MS m/z: 587.4 [M+H].sup.+

Embodiment 60: Compound 60

(414) ##STR00256##

(415) ##STR00257##

Step 1: Synthesis of Compound 60-1

(416) Cesium carbonate (295.82 mg, 907.94 mol) and 1-bromo-2-fluoroethane (92.22 mg, 726.34 mol) were added into the compound 12-E (100.00 mg, 363.17 mol) in DMF solution (4.00 mL), the reaction was stirred at 80 C. for 1 hour, 10 mL water was added into the reaction mixture, the aqueous phase was washed by EA (15 mL3), the organic phases were combined and washed by saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, the crude product was purified by column chromatography (dichloromethane/methanol=10/1) to give the 60-1. MS m/z: 322.1 [M+H].sup.+

Step 2: Synthesis of Compound 60-2

(417) Ammonium chloride (183.08 mg, 3.42 mmol) and zinc (179.04 mg, 2.74 mmol) were added into the compound 60-1 (110.00 mg, 342.26 mol) in ethanol (20.00 mL) and water (2.5.00 mL), the reaction was stirred at 70 C. for 1 hour, the reaction mixture was filtered, the filtrate was concentrated to give a crude product 60-2. MS m/z: 292.1 [M+H].sup.+

Step 3: Synthesis of Compound 60

(418) Compound 60 was obtained by using the same methods as preparing the compound 22 in embodiment 22, except for the corresponding starting material.

(419) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.60 (s, 6H) 1.63-1.70 (m, 8H) 2.55 (br t, J=5.14 Hz, 4H) 2.71 (t, J=5.02 Hz, 1H) 2.78 (t, J=4.88 Hz, 1H) 3.14-3.19 (m, 4H) 4.00 (br s, 1H) 4.55 (t, J=4.8 Hz, 1H) 4.67 (t, J=4.8 Hz, 1H) 4.76 (d, J=6.4 Hz, 2H) 4.89-5.03 (m, 1H) 5.06 (d, J=10.0 Hz, 1H) 5.72 (ddt, J=16.82, 10.42, 6.22, 6.22 Hz, 1H) 6.95 (d, J=9.02 Hz, 2H) 7.35 (d, J=7.52 Hz, 1H) 7.47 (br d, J=8.52 Hz, 2H) 7.78 (d, J=8.02 Hz, 1H) 7.88 (t, J=7.92 Hz, 1H) 8.85 (s, 1H). MS m/z: 601.1 [M+H].sup.+

Embodiment 61: Compound 61

(420) ##STR00258##

(421) Synthetic Route:

(422) ##STR00259##

Step 1: Synthesis of Compound 61-A

(423) Cesium carbonate (295.82 mg, 907.94 mol) and 2-bromo-1,1-difluoroethane (52.64 mg, 363.17 mol) were added into the compound 12-E (100.00 mg, 363.17 mol) in DMF solution (4.00 mL), the reaction was stirred at 80 C. for 1 hour, the reaction was stirred at 85 C. for further 12 hours. Then 10 mL water was added into the reaction mixture, the aqueous phase was washed by EA (15 mL3), the organic phases were combined and washed by saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, the crude product was separated by column chromatography (PE/EA=1/1) to give the compound 61-1. MS m/z: 340.1 [M+H].sup.+

Step 2: Synthesis of Compound 61-2

(424) Ammonium chloride (295.82 mg, 907.94 mol) and zinc (52.41 mg, 801.44 mol) were added into the compound 61-1 (34.00 mg, 100.18 mol) in ethanol (10.00 mL) and water (1.00 mL), the reaction was stirred at 70 C. for 1 hour, the reaction mixture was filtered and the filtrate was concentrated to give a crude product 61-2. MS m z: 310.1 [M+H].sup.+

Step 3: Synthesis of Compound 61

(425) Compound 61 was obtained by using the same methods as preparing the compound 22 in embodiment 22, except for the corresponding starting material.

(426) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.52 (s, 6H) 1.52-1.63 (m, 8H) 2.48-2.52 (m, 4H) 2.69 (td, J=15.06, 4.52 Hz, 2H) 3.06-3.09 (m, 4H) 3.89 (br s, 1H) 4.67 (d, J=6.02 Hz, 2H) 4.87 (dd, J=17.08, 1.00 Hz, 1H) 4.97 (dd, J=10.04, 1.00 Hz, 1H) 5.58-5.70 (m, 1H) 5.80-5.99 (m, 1H) 6.86 (d, J=9.02 Hz, 2H) 7.27 (d, J=7.52 Hz, 1H) 7.38 (br d, J=8.52 Hz, 2H) 7.69 (d, J=8.02 Hz, 1H) 7.76-7.81 (m, 1H) 8.75 (s, 1H). MS m/z: 619.1 [M+H].sup.+

Embodiment 62: Compound 62

(427) ##STR00260##

(428) Synthetic Route:

(429) ##STR00261##

Step 1: Synthesis of Compound 62-1

(430) The compound 62-A was obtained by using the same methods as preparing the compound 60-1 in embodiment 60, except for the corresponding starting material. MS m/z: 329.1 [M+H].sup.+

Step 2: Synthesis of Compound 62-2

(431) Wet palladium carbon (28.74 mg, 24.36 mol, 10% purity) was added in the compound 62-1 (80.00 mg, 243.60 mol) in THF solution (10.00 mL), the reaction was stirred at 20 C. for 12 hours under hydrogen (15 psi) atmosphere, the reaction mixture was filtered and the filtrate was concentrated to give a crude product 62-2. MS m/z: 299.2 [M+H].sup.+

Step 3: Synthesis of Compound 62

(432) The compound 62 was obtained by using the same methods as preparing the compound 22 in embodiment 22, except for the corresponding starting material. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.52 (s, 6H) 1.56-1.60 (m, 8H) 2.41-2.45 (m, 4H) 2.45-2.50 (m, 2H) 2.63-2.68 (m, 2H) 3.05-3.09 (m, 4H) 3.89 (br s, 1H) 4.67 (br d, J=6.02 Hz, 2H) 4.87 (d, J=17.82 Hz, 1H) 4.97 (d, J=10.54 Hz, 1H) 5.56-5.71 (m, 1H) 6.86 (d, J=8.78 Hz, 2H) 7.27 (d, J=7.52 Hz, 1H) 7.38 (br d, J=8.02 Hz, 2H) 7.69 (d, J=8.02 Hz, 1H) 7.76-7.82 (m, 1H) 8.76 (s, 1H). MS m/z: 608.1 [M+H].sup.+

Embodiment 63: Compound 63

(433) ##STR00262##

(434) Synthetic Route:

(435) ##STR00263##

Step 1: Synthesis of Compound 63-1

(436) The compound 63-1 was obtained by using the same methods as preparing the compound 60-1 in embodiment 60, except for the corresponding starting material. MS m/z: 315.1 [M+H].sup.+

Step 2: Synthesis of Compound 63-2

(437) The compound 63-2 was obtained by using the same methods as preparing the compound 62-2 in embodiment 62, except for the corresponding starting material. MS m/z: 285.1 [M+H].sup.+

Step 3: Synthesis of Compound 63

(438) The compound 63 was obtained by using the same methods as preparing the compound 22 in embodiment 22, except for the corresponding starting material. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.51 (s, 6H) 1.54-1.57 (m, 4H) 1.58-1.63 (m, 4H) 2.49-2.55 (m, 4H) 3.04-3.11 (m, 4H) 3.48 (s, 2H) 3.87 (s, 1H) 4.66 (d, J=6.02 Hz, 2H) 4.86 (d, J=17.08 Hz, 1H) 4.96 (d, J=10.04 Hz, 1H) 5.57-5.68 (m, 1H) 6.85 (d, J=9.02 Hz, 2H) 7.24-7.28 (m, 1H) 7.38 (br d, J=8.52 Hz, 2H) 7.68 (d, J=8.02 Hz, 1H) 7.76-7.81 (m, 1H) 8.75 (s, 1H). MS m/z: 594.1 [M+H].sup.+

Embodiment 64: Compound 64

(439) ##STR00264##

(440) Synthetic Route:

(441) ##STR00265##

Step 1: Synthesis of Compound 64-1

(442) The compound 64-1 was obtained by using the same methods as preparing the compound 60-1 in embodiment 60, except for the corresponding starting material. MS m/z: 334.2[M+H].sup.+

Step 2: Synthesis of Compound 64-1

(443) The compound 64-1 was obtained by using the same methods as preparing the compound 61-1 in embodiment 61, except for the corresponding starting material. MS m/z: 304.1 [M+H].sup.+

Step 3: Synthesis of Compound 64

(444) The compound 64 was obtained by using the same methods as preparing the compound 22 in embodiment 22, except for the corresponding starting material. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.52 (s, 6H) 1.52-1.62 (m, 8H) 2.41 (br s, 4H) 2.53 (t, J=5.78 Hz, 2H) 3.05-3.10 (m, 4H) 3.29 (s, 3H) 3.46 (t, J=5.66 Hz, 2H) 3.92 (br s, 1H) 4.67 (br d, J=6.02 Hz, 2H) 4.87 (dd, J=17.08, 1.00 Hz, 1H) 4.97 (d, J=10.04 Hz, 1H) 5.63 (ddt, J=16.86, 10.38, 6.22, 6.22 Hz, 1H) 6.86 (d, J=9.04 Hz, 2H) 7.27 (d, J=7.54 Hz, 1H) 7.38 (br d, J=8.54 Hz, 2H) 7.69 (d, J=8.04 Hz, 1H) 7.76-7.82 (m, 1H) 8.75 (s, 1H). MS m/z: 613.1[M+H].sup.+

Embodiment 65: Compound 65

(445) ##STR00266##

(446) Synthetic Route:

(447) ##STR00267##

Step 1: Synthesis of Compound 65-1

(448) n-Butyllithium (2.5 M, 371.48 L) was added into 2,6-dibromopyridine (200 mg, 844.27 mol) in 6 mL DCM suspension at 60 C. The reaction mixture was stirred for 15 min, 3,3-difluorocyclobutanone (134.71 mg, 1.27 mmol) was added into the reaction mixture by one time at 60 C. The reaction mixture was stirred at 60 C. for further 60 min. The reaction mixture was poured into saturated ammonium chloride, the organic phase was separated, washed by brine 10 mL, dried over anhydrous sodium sulfate, concentrated under reduced pressure, separated and purified by silica gel column chromatography (PE/EA=5/1) to give the compound 65-1. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 7.57 (t, J=8.0 Hz, 1H) 7.48 (d, J=7.6 Hz, 1H) 7.39 (d, J=8.0 Hz, 1H) 4.48 (s, 1H) 2.98-3.08 (m, 4H)

Step 2: Synthesis of Compound 65-2

(449) At 10 C. and under nitrogen atmosphere, DAST (351.39 mg, 2.18 mmol) was added rapidly dropwise into 65-1 (250.00 mg, 1.09 mmol) in DCM (10.00 mL) solution. Then the mixture was stirred at 10 C. for 60 min under nitrogen atmosphere. The reaction was quenched by saturated sodium bicarbonate solution 10 mL, then extracted by DCM (20 mL2), the combined organic phases were dried over anhydrous sodium sulfate, then filtered and concentrated to give 65-2 crude product, which was used directly in the next step. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 7.53 (t, J=8.0 Hz, 1H) 7.39-7.43 (m, 2H) 5.05-5.13 (m, 2H) 4.87-4.95 (m, 2H)

Step 3: Synthesis of Compound 65-3

(450) N,N-dimethylethylenediamine (18.76 mg, 212.81 mol) was added into I1 (43.00 mg, 193.46 mol), cuprous iodide (36.84 mg, 193.46 mol), 65-2 (51.47 mg, 193.46 mol) and potassium carbonate (37.43 mg, 270.84 mol) in dioxane (3.00 mL) solution, the mixture was stirred at 95 C. for 2 hours under nitrogen atmosphere. 10 mL ammonia was added into the mixture and extracted with EA (15 mL2), the combined organic phases were washed by saturated brine, dried over anhydrous magnesium sulfate, then filtered and concentrated to give the crude product 65-3 (yellow oily product, 60.00 mg), which was used directly in the next step. MS m/z: 408.0 [M+H].sup.+

Step 4: Synthesis of Compound 65

(451) m-CPBA (70.64 mg, 347.93 mol, 85%) were added into 65-3 (105.00 mg, 257.73 mol) in toluene (5.00 mL) solution at 20-25 C., the mixture was stirred at 20-25 C. for 60 min, then DIEA (91.60 mg, 708.75 mol) and 12 (66.85 mg, 257.73 mol) were added below 30 C., the mixture was stirred at 20-25 C. for 16 hours. Then the mixture was diluted by 40 mL EA, and washed sequentially by 20 mL saturated sodium sulfite solution, saturated sodium carbonate 20 mL and brine 20 mL, dried over anhydrous sodium sulfate, concentrated under reduced pressure to give a crude product. The crude product was purified by high performance liquid phase (alkaline condition) and thin layer chromatography (DCM/MeOH=10/1) to give the compound 65. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 8.76 (s, 1H) 7.88 (d, J=8.0 Hz, 1H) 7.82 (t, J=8.0 Hz, 1H) 7.36-7.40 (m, 3H) 6.86 (d, J=8.8 Hz, 2H) 5.56-5.66 (m, 1H) 4.83-4.97 (m, 2H), 4.74 (d, J=6.4 Hz, 2H) 3.31-3.39 (m, 2H) 3.07-3.17 (m, 6H) 2.45 (br, 4H) 2.30 (s, 3H) 1.59-1.52 (m, 8H) MS m/z/619.0 [M+H].sup.+

Embodiment 66: Compound 66

(452) ##STR00268##

(453) Synthetic Route:

(454) ##STR00269##

Step 1: Synthesis of Compound 66-1

(455) The compound 66-1 was obtained by using the same methods as preparing the compound 46-A in embodiment 46, except for the corresponding starting material. MS m/z: 406.0 [M+H].sup.+

Step 2: Synthesis of Compound 66

(456) The compound 66 was obtained by using the same methods as preparing the compound 22 in embodiment 22, except for the corresponding starting material.

(457) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.58-1.70 (m, 8H) 2.32 (s, 3H) 2.42 (br s, 4H) 3.09-3.23 (m, 8H) 4.72 (br d, J=6.52 Hz, 2H) 4.98 (dd, J=17.06, 1.00 Hz, 1H) 5.06-5.10 (m, 1H) 5.73 (ddt, J=16.82, 10.42, 6.22, 6.22 Hz, 1H) 6.94 (d, J=9.02 Hz, 2H) 7.44 (br d, J=8.52 Hz, 2H) 7.54 (d, J=7.54 Hz, 1H) 7.86 (d, J=8.04 Hz, 1H) 7.92-7.97 (m, 1H) 8.83 (s, 1H). MS m/z: 617.1[M+H].sup.+

Embodiment 67: Synthesis of Compound 67

(458) ##STR00270##

(459) Synthetic Route:

(460) ##STR00271##

Step 1: Synthesis of Compound 67-1

(461) The compound 67-1 was obtained by using the same methods as preparing the compound 46-A in embodiment 46, except for the corresponding starting material. MS m/z: 342.1 [M+H].sup.+

Step 2: Synthesis of Compound 67

(462) The compound 67 was obtained by using the same methods as preparing the compound 22 in embodiment 22, except for the corresponding starting material. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.24 (s, 3H) 1.26 (s, 3H) 1.50-1.61 (m, 8H) 2.23 (s, 3H) 2.33 (br s, 4H) 2.96-3.04 (m, 1H) 3.04-3.11 (m, 4H) 4.78-4.85 (m, 3H) 4.92 (d, J=10.04 Hz, 1H) 5.61 (ddt, J=16.76, 10.36, 6.54, 6.54 Hz, 1H) 6.85 (d, J=9.04 Hz, 2H) 7.00 (d, J=7.02 Hz, 1H) 7.39 (br d, J=8.52 Hz, 2H) 7.60-7.69 (m, 2H) 8.74 (s, 1H). MS m/z: 553.1[M+H].sup.+

Embodiment 68: Synthesis of Compound 68

(463) ##STR00272##

(464) Synthetic Route:

(465) ##STR00273##

Step 1: Synthesis of Compound 68-1

(466) Potassium isopropenyltrifluoroborate (1.12 g, 7.60 mmol) were added into 2,6-dibromopyridine (1.50 g, 6.33 mmol) in ethanol (20 mL) solution in a sealed tube. After nitrogen bubbling for 10 min, Pd (dppf)Cl.sub.2.Math.CH.sub.2Cl.sub.2 (258.47 mg, 316.50 mol) and triethylamine (640.53 mg, 6.33 mmol, 877.44 L) were added into the mixture. The sealed tube was heated to 85 C., then stirred for 4.5 hours. After cooling cooled down to r.t., the mixture was concentrated to dryness, then water 50 mL and EA 30 mL were added, partitioned, the aqueous phase was extracted by EA (20 mL2), the combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column machine (PE/EA=100/0-30/1) to give the compound 68-1. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 7.28-7.44 (m, 3H) 5.86 (s, 1H) 5.22-5.25 (m, 1H) 2.10 (s, 3H)

Step 2: Synthesis of Compound 68-2

(467) Potassium tert-butoxide (679.85 mg, 6.06 mmol) was added into trimethyl iodine sulfoxide (1.33 g, 6.06 mmol) in DMSO (9.2 mL) and THF (6.00 mL) solution at 15-20 C. by one time. The mixture was stirred for 30 min, then 68-1 (800.00 mg, 4.04 mmol) in THF solution (9.20 mL) solution was added at the same temperature. The mixture was stirred for more than 120 min at 15-20 C., then stirred for 1 hour at 60 C. The reaction was quenched by 10 mL water, then diluted by 50 mL EA. The organic phase was washed by water (20 mL3) and brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. The crude product was purified by thin layer chromatography (PE/EA=50/1) to give the crude product compound 68-2. MS m/z: 211.8 [M+H].sup.+

Step 3: Synthesis of Compound 68-3

(468) N,N-dimethyllethylenediamine (40.85 mg, 463.40 mol) was added into I1 (103.00 mg, 463.40 mol), cuprous iodide (88.25 mg, 463.40 mol), 68-2 (184.74 mg, 463.40 mol) and potassium carbonate (64.05 mg, 463.40 mol) in dioxane (4.00 mL) solution, the mixture was stirred at 95 C. for 2 hours under nitrogen atmosphere. 10 mL ammonia was added and the mixture was extracted by EA (15 mL2), the combined organic phases were washed by saturated brine, dried over anhydrous sodium sulfate, then filtered and concentrated to give a red oily product. The product was purified by thin layer chromatography (PE/EA=3/1) to give 68-3. MS m/z: 354.1 [M+H].sup.+

Step 4: Synthesis of Compound 68

(469) m-CPBA (15.51 mg, 76.40 mol, 85%) was added into 68-3 (20.00 mg, 56.59 mol) in toluene (5.00 mL) solution at 20-25 C., the mixture was stirred at 20-25 C. for 60 min, then DIEA (20.11 mg, 155.62 mol) and 12 (14.68 mg, 56.59 mol) were added below 30 C., the mixture was stirred at 20-25 C. for 16 hours. The mixture was diluted by 40 mL EA, then washed sequentially by 20 mL saturated sodium sulfite solution, saturated sodium carbonate 20 mL and brine 20 mL, dried over anhydrous sodium sulfate, concentrated under reduced pressure to give a crude product. The crude product was purified by preparative thin layer chromatography (DCM/MeOH=10/1) twice to give the compound 68. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 8.74 (s, 1H), 7.66 (t, J=8.0 Hz, 1H) 7.54 (d, J=8.0 Hz, 1H) 7.39 (d, J=8.0 Hz, 2H) 7.12 (d, J=8.0 Hz, 1H) 6.84 (d, J=8.8 Hz, 2H) 5.56-5.64 (m, 1H) 4.81-4.95 (m, 2H), 4.71 (d, J=6.0 Hz, 2H) 3.08-3.09 (m, 4H) 2.59 (br, 4H) 2.40 (s, 3H) 1.62-1.68 (m, 8H) 1.42 (s, 3H) 0.77-0.81 (m, 4H) MS m/z/565.1 [M+H].sup.+

Embodiment 69: Compound 69

(470) ##STR00274##

(471) Synthetic Route:

(472) ##STR00275##

Step 1: Synthesis of Compound 69-1

(473) 3-Oxacyclobutanone (23.55 mg, 326.85 mol) was added into the compound 12-E (60.00 mg, 217.90 mol) in DCM (4.00 mL) solution, the reaction mixture was stirred at 20 C. for 30 min, then sodium triacetylborohydride (92.36 mg, 435.80 mol) was added, the reaction mixture was stirred at 25 C. for 30 min. The reaction mixture adjusted to pH=5-6 by adding hydrochloric acid, then adjusted to pH=9-10 by adding sodium hydroxide solution, and then extracted by DCM (10 mL3), the organic phases were combined and washed by saturated brine (15 mL1), dried over anhydrous sodium sulfate, then filtered, the filtrate was evaporated to dry to give the crude product 69-1. MS m/z: 332.2 [M+H].sup.+

Step 2: Synthesis of Compound 69-2

(474) The compound 69-2 was obtained by using the same methods as preparing the compound 61-2 in embodiment 61, except for the corresponding starting material. MS m/z: 302.1 [M+H].sup.+

Step 3: Synthesis of Compound 69

(475) The compound 69 was obtained by using the same methods as preparing the compound 22 in embodiment 22, except for the corresponding starting material. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.52 (s, 6H) 1.53-1.62 (m, 8H) 2.22 (br s, 4H) 3.03-3.12 (m, 4H) 3.42 (quin, J=6.54 Hz, 1H) 3.91 (br s, 1H) 4.54-4.62 (m, 4H) 4.67 (br d, J=6.02 Hz, 2H) 4.83-4.94 (m, 1H) 4.97 (d, J=10.54 Hz, 1H) 5.63 (ddt, J=16.82, 10.42, 6.22, 6.22 Hz, 1H) 6.86 (d, J=8.54 Hz, 2H) 7.27 (d, J=7.54 Hz, 1H) 7.38 (br d, J=8.04 Hz, 2H) 7.67-7.71 (m, 1H) 7.76-7.81 (m, 1H) 8.75 (s, 1H). MS m/z: 611.1 [M+H].sup.+

Embodiment 70: Compound 70

(476) ##STR00276##

(477) Synthetic Route

(478) ##STR00277##

Step 1: Synthesis of Compound 70-1

(479) At 0-5 C., oxalyl chloride (629.57 mg, 4.96 mmol, 434.19 L) was added into 6-bromo-2-pyridinecarboxylic acid (500.00 mg, 2.48 mmol) in DCM (10.00 mL) solution, followed by adding DMF (181.26 mg, 2.48 mmol, 190.80 L). The mixture was stirred at 0-5 C. for 0.5 hours, and then stirred at 25-35 C. for 1.5 hours, then concentrated under reduced pressure to give the yellow solid. Dimethylamine solution (2 M, 3.25 mmol, 11.38 mL) was slowly added into said yellow solid in THF solution (3.00 mL) at 0 C., the reaction mixture was stirred at 25 C. for 3 hours. The reaction mixture was concentrated up to dryness, then diluted by water 20 mL, the aqueous phase was extracted by DCM (10 mL3), the organic phase was washed by 20 mL saturated brine, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated up to dryness to give the crude product 70-1. MS m/z: 230.9 [M+H].sup.+

Step 2: Synthesis of Compound 70-2

(480) Cuprous iodide (362.41 mg, 1.90 mmol), N,N-dimethylethylenediamine (187.87 mg, 2.13 mmol, 229.11 L) and potassium carbonate (362.94 mg, 2.63 mmol) were added separately into the compound I1 (422.96 mg, 1.90 mmol) and 70-1 (450.00 mg, 1.96 mmol) in dioxane (5 mL) solution. The reaction mixture was stirred at 95 C. for 1 hour under nitrogen atmosphere then concentrated, 20 mL ammonia was added after concentration, then extracted by EA 150 mL (50 mL3) and washed by saturated brine 30 mL, dried over anhydrous sodium sulfate, then filtered to give the crude compound. The crude product was purified by silica gel chromatography (PE/EA=1/1-0/1) to give the 70-2. MS m/z: 371.0 [M+H].sup.+

Step 3: Synthesis of Compound 70

(481) m-CPBA (67.29 mg, 331.44 mol, 85% purity) was added into the compound 70-2 (111.10 mg. 299.93 mol) in toluene (8.00 mL) solution at 35-40 C., and stirred at 25-30 C. for 0.5 hours, then the compound 13 (91.00 mg, 299.93 mol) and DIPEA (91.00 mg, 299.93 mol, 157.15 L) were added into the mixture, and stirred for further 16 hours. The reaction mixture was diluted by DCM 45 mL, washed by saturated sodium bicarbonate 15 mL, dried over sodium sulfate, then filtered and concentrated to give the crude compound, the crude compound was purified by preparative HPLC (chromatographic column: Xtimate C18 15025 mm5 m; mobile phases: [water (0.05% ammonia hydroxide v/v)-ACN]; B (acetonitrile) %: 30%-60%, 10 min) to give the 70. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.76 (s, 1H), 7.92-7.83 (m, 2H), 7.48 (d, J=8.1 Hz, 1H), 7.37 (d, J=9.2 Hz, 2H), 6.87 (d, J=9.2 Hz, 2H), 5.63-5.56 (m, 1H), 4.98-4.85 (m, 2H), 4.69 (d, J=6.4 Hz, 2H), 3.63 (s, 3H), 3.40 (br, 4H), 3.13-3.07 (m, 7H), 3.02 (s, 3H), 1.65-1.59 (m, 4H), 1.44 (br, 4H). MS m/z/626.1 [M+H].sup.+

Embodiment 71: Synthesis of Compound 71

(482) ##STR00278##

(483) ##STR00279##

Step 1: Synthesis of Compound 71-1

(484) Cuprous iodide (413.91 mg, 2.17 mmol), N,N-dimethyllethylenediamine (214.57 mg, 2.43 mmol, 261.67 L) and potassium carbonate (414.52 mg, 3.00 mmol) were added separately into the compound I1 (483.07 mg, 2.17 mmol) and 42-A (500.00 mg, 2.17 mmol) in dioxane (5 mL) solution. The reaction mixture was stirred at 95 C. for 1 hour under nitrogen atmosphere, then concentrated and 20 mL ammonia was added, extracted by EA 150 mL (50 mL3) and washed by saturated brine 30 mL, dried over anhydrous sodium sulfate, then filtered to give the crude compound. The crude product was purified by column chromatography (PE/EA=1/1) to give the 71-1. MS m/z: 372.0 [M+H].sup.+

Step 2: Synthesis of Compound 71

(485) m-CPBA (71.06 mg, 350.01 mol, 85% purity) was added into the compound 71-1 (100.00 mg, 269.24 mol) in toluene (6.00 mL) solution at 35-40 C., then the mixture was stirred at 25-30 C. for 0.5 hours and then the compound 13 (81.69 mg, 269.24 mol) and DIPEA (104.39 mg, 807.72 mol, 141.07 L) were added, then stirred for further 16 hours. The reaction mixture was diluted by DCM 45 mL, washed by saturated sodium bicarbonate 15 mL, dried over sodium sulfate, then filtered and concentrated to give the crude compound, the crude compound was purified by preparative HPLC (column: YMC-Actus Triart C18 15030 mm 5 m; mobile phases: [water (0.05% HCl)-ACN]; B (acetonitrile) %: 25%-55%, 9 min) to give the 71. .sup.1H NMR (400 MHz, CDCl.sub.3) =8.83 (s, 1H), 8.07-8.00 (m, 1H), 7.88 (dd, J=7.8, 13.6 Hz, 2H), 7.45 (br d, J=8.5 Hz, 2H), 6.94 (d, J=9.0 Hz, 2H), 5.72 (tdd, J=6.0, 10.4, 16.8 Hz, 1H), 5.26 (s, 1H), 5.13-5.06 (m, 3H), 4.97 (d, J=17.1 Hz, 1H), 4.78 (d, J=7.0 Hz, 2H), 4.65 (br d, J=6.0 Hz, 2H), 3.71 (s, 3H), 3.49 (br d, J=6.0 Hz, 4H), 3.23-3.13 (m, 4H), 1.72-1.68 (m, 4H), 1.52 (br s, 4H). MS m/z: 627.1[M+H].sup.+

Embodiment 72: Compound 72

(486) ##STR00280##

(487) Synthetic Route:

(488) ##STR00281##

Step 1: Synthesis of Compound 72-1

(489) Cesium carbonate (413.91 mg, 2.17 mmol), tris(dibenzylideneacetone)dipalladium (21.21 mg, 23.16 mol) and 4,5-bisdiphenylphosphino-9,9-dimethyloxazepine (26.80 mg, 46.32 mol) were added separately into the compound 3,3-difluoroazetidine hydrochloride (30.00 mg, 231.59 mol) and 2,6-dibromopyridine (54.86 mg, 231.59 mol) in dioxane solution (1 mL), the reaction mixture was stirred at 90 C. for 16 hours under nitrogen atmosphere, then concentrated and added 10 mL water, extracted by DCM 30 mL (10 mL3) and washed by saturated brine 20 mL, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by thin layer chromatography (PE/EA=10/1) to give the 72-1. MS m/z: 250.9[M+H].sup.+

Step 2: Synthesis of Compound 72-2

(490) Cuprous iodide (42.82 mg, 224.85 mol), N,N-dimethyllethylenediamine (22.20 mg, 251.83 mol, 27.07 L) and potassium carbonate (42.88 mg, 310.29 mol) were added separately into the compound I1 (49.98 mg, 224.85 mol) and 72-1 (56.00 mg, 224.85 mol) in dioxane (3 mL) solution, the reaction mixture was stirred at 95 C. for 1 hour under nitrogen atmosphere, then concentrated and added 20 mL ammonia, extracted by EA 150 mL (50 mL3) and washed by saturated brine 30 mL, dried over anhydrous sodium sulfate, then filtered to give the crude compound. The crude product was purified by silica gel column chromatography (PE/EA=3/1) 72-1. MS m/z: 391.0 [M+H].sup.+

Step 3: Synthesis of Compound 72

(491) m-CPBA (22.51 mg, 110.88 mol, 85% purity) was added into the compound 72-2 (33.30 mg, 85.29 mol) in toluene (3.00 mL) solution at 35-40 C., the mixture was stirred at 25-30 C. for 0.5 hours and then the compound 12 (22.12 mg, 85.29 mol) and DIPEA (33.07 mg, 255.87 mol, 44.69 L) were added, then stirred for further 16 hours. The reaction mixture was diluted by EA 45 mL, then washed by saturated sodium bicarbonate 15 mL, dried over sodium sulfate, then filtered and concentrated to give the crude compound. The other batch employed the same method and used 12 7.7 mg to give a batch of crude product. The two batches of crude product was combined and purified by preparative HPLC (chromatographic column: YMC-Actus Triart C18 15030 mm 5 m; mobile phases: [water (0.05% HCl)-ACN]; B (acetonitrile) %: 15%-45%, 9 min) to give the 72. .sup.1H NMR (400 MHz, METHANOL-d.sub.4) 6=8.67 (s, 1H), 7.68 (t, J=8.0 Hz, 1H), 7.45 (d, J=8.4 Hz, 2H), 7.11 (d, J=8.0 Hz, 1H), 6.67-6.84 (m, 2H), 6.39 (d, J=8.8 Hz, 1H), 5.66-5.60 (m, 1H), 4.97 (d, J=10 Hz, 1H), 4.88 (d, J=15.6 Hz, 1H), 4.62 (d, J=6.0 Hz, 2H), 4.30 (t, J=12 Hz, 4H), 3.02-2.99 (m, 4H), 2.41 (br, 4H), 2.23 (s, 3H), 1.57-1.49 (m, 8H) MS m/z: 602.0[M+H].sup.+

Embodiment 73: Compound 73

(492) ##STR00282##

(493) Synthetic Route

(494) ##STR00283##

Step 1: Synthesis of Compound 73-1

(495) Sodium hydride (3.48 g, 86.93 mmol, 60% purity) was added into the compound 42-A (5 g, 21.73 mmol) in THF solution (3 mL) at 0 C., and then methyl iodide (10.6 g, 74.68 mmol, 4.65 mL) was added, the reaction mixture was stirred at 10 C. for 16 hours. Saturated ammonium chloride solution (30 mL) was added into the reaction mixture and the mixture was extracted with EA (20 mL2), then washed by saturated brine 20 mL, dried over sodium sulfate, then filtered and concentrated to give 73-1. .sup.1H NMR (400 MHz, CDCl.sub.3) =7.43 (d, J=8.0 Hz, 1H), 7.29 (d, J=8.0 Hz, 1H), 7.23 (d, J=6.4 Hz, 1H), 4.88 (d, J=7.6 Hz, 2H), 4.69 (d, J=6.0 Hz, 2H), 3.08 (s, 3H)

Step 2: Synthesis of Compound 73-2

(496) Cuprous iodide (4.11 g, 21.60 mmol), N,N-dimethyllethylenediamine (2.13 g, 24.19 mmol, 2.60 mL) and potassium carbonate (4.12 g, 29.80 mmol) were added separately into the compound I1 (4.8 g, 21.60 mmol) and 73-1 (5.27 g, 21.60 mmol) in dioxane (80 mL) solution, the reaction mixture was stirred at 95 C. for 1 hour under nitrogen atmosphere. Then the mixture was cooled down, 40 mL ammonia was added and extracted by EA (50 mL3), then washed by saturated brine 50 mL, dried over anhydrous sodium sulfate, then filtered to give the crude compound. The crude product was purified by column chromatography (PE/EA=3/1-1/1) to give the 73-2. MS m/z: 386.0 [M+H].sup.+

Step 3: Synthesis of Compound 73

(497) m-CPBA (2.40 g, 11.82 mmol, 85% purity) was added into the compound 73-2 (3.4 g, 8.82 mmol) in toluene (40 mL) solution at 35-40 C., the mixture was stirred at 20 C. for 1 hour and then the compound DIPEA (3.42 g, 26.46 mmol, 4.61 mL) and 12 (2.40 g, 9.26 mmol) were added, then stirred at 20 C. for further 12 hours. 30 mL water was added, the mixture was extracted by EtOAc (40 mL3), the organic phases were combined and washed sequentially by saturated sodium bicarbonate 40 mL, saturated brine 30 mL, dried over sodium sulfate, then filtered and concentrated to give the crude compound. The crude compound was purified by preparative HPLC (chromatographic column: Phenomenex luna C18 25050 mm10 m; mobile phases: [water (0.1% TFA)-ACN]; B (acetonitrile) %: 5%-30%, 23 min) to give the 73. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.69 (br dd, J=11.54, 6.02 Hz, 8H) 2.38 (s, 3H) 2.50 (br s, 4H) 3.13-3.22 (m, 4H) 3.28 (s, 3H) 4.91-4.97 (m, 5H) 5.01-5.06 (m, 3H) 5.66-5.76 (m, 1H) 6.96 (d, J=9.04 Hz, 2H) 7.35-7.39 (m, 1H) 7.48 (br d, J=9.04 Hz, 2H) 7.88-7.98 (m, 2H) 8.86 (s, 1H). MS m/z: 597.1 [M+H].sup.+

Embodiment 74: Synthesis of Compound 74, 75

(498) ##STR00284##

(499) Synthetic Route:

(500) ##STR00285##

Step 2: Synthesis of Compound 74-1

(501) 2-Bromo-6-acetyl pyridine (2 g, 10.00 mmol) was added into the white suspension of the compound sodium acetate (820.21 mg, 10.00 mmol) in dimethyl sulfoxide (12 mL), then 2 mL dimethyl sulfoxide was added, trifluoromethyl trimethylsilane (5.69 g, 39.99 mmol) was slowly added dropwise into the reaction mixture at 10-20 C., the reaction mixture was stirred at 20 C. for 12 hours. The reaction mixture was placed in an ice bath, the inner temperature was kept between 10-25 C., 16 mL water was added into the reaction mixture to quench the reaction, the aqueous phase was extracted by EA (36 mL3), the organic phases were combined and washed by saturated sodium bicarbonate 40 mL, and the organic phase was then washed by saturated brine 40 mL, dried over anhydrous sodium sulfate, then filtered, the filtrate was evaporated to give the crude product, which was purified by silica gel column chromatography (PE/EA=15/1) to give 74-1.

(502) MS m/z: 269.9[M+H].sup.+

Step 2: Synthesis of Compound 74-2

(503) The compound 74-2 was obtained by using the same methods as preparing the compound 46-A in embodiment 46, except for the corresponding starting material. MS m/z: 412.0 [M+H].sup.+

Step 3: Synthesis of Compound 74-3

(504) The compound 74-3 was obtained by using the same methods as preparing the compound 22 in embodiment 22, except for the corresponding starting material.

(505) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.52-1.61 (m, 8H) 1.70 (s, 3H) 2.23 (s, 3H) 2.34 (br s, 4H) 3.05-3.12 (m, 4H) 4.50 (dd, J=15.56, 7.04 Hz, 1H) 4.71 (dd, J=15.56, 6.02 Hz, 1H) 4.86 (d, J=18.06 Hz, 1H) 4.98 (d, J=9.54 Hz, 1H) 5.62 (ddt, J=16.76, 10.48, 6.22, 6.22 Hz, 1H) 6.87 (d, J=9.04 Hz, 2H) 7.32-7.42 (m, 3H) 7.88 (d, J=3.52 Hz, 2H) 8.76 (s, 1H). MS m/z: 623.1[M+H].sup.+

Step 4: Synthesis of Compound 74, 75

(506) Compound 74-3 was separated by SFC (chiral column: CHIRALCELR OJ-H (Particle Size: 5 m Dimensions: 30 mm 250 mm) DAICEL CHEMICAL INDUSTRIES, LTD. mobile phases: A: CO.sub.2, B: EtOH (0.10% NH.sub.3H.sub.2O), A:B=65:35, flow rate: 50 mL/min) to give 74 (7.00 min). .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.52 (br t, J=5.52 Hz, 4H) 1.58-1.61 (m, 4H) 1.70 (s, 3H) 2.23 (s, 3H) 2.33 (br s, 4H) 3.07-3.11 (m, 4H) 4.50 (dd, J=15.81, 6.78 Hz, 1H) 4.71 (dd, J=15.81, 5.77 Hz, 1H) 4.86 (dd, J=17.07, 1.00 Hz, 1H) 4.98 (d, J=9.54 Hz, 1H) 5.39 (br s, 1H) 5.56-5.67 (m, 1H) 6.87 (d, J=9.03 Hz, 2H) 7.36 (br d, J=7.53 Hz, 3H) 7.87-7.91 (m, 2H) 8.76 (s, 1H). MS m/z: 623.1 [M+H].sup.+ and 75 (5.55 min). .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.52 (br t, J=5.52 Hz, 4H) 1.57-1.61 (m, 4H) 1.70 (s, 3H) 2.23 (s, 3H) 2.33 (br s, 4H) 3.04-3.13 (m, 4H) 4.50 (dd, J=15.81, 6.78 Hz, 1H) 4.71 (dd, J=15.56, 5.52 Hz, 1H) 4.86 (dd, J=17.07, 1.00 Hz, 1H) 4.98 (d, J=9.54 Hz, 1H) 5.38 (br s, 1H) 5.56-5.67 (m, 1H) 6.87 (d, J=9.03 Hz, 2H) 7.33-7.39 (m, 3H) 7.89 (d, J=3.51 Hz, 2H) 8.77 (s, 1H). MS m/z: 623.1[M+H].sup.+

Embodiment 75: Synthesis of Compound 76

(507) ##STR00286##

(508) Synthetic Route:

(509) ##STR00287##

Step 1: Synthesis of Compound 76-1

(510) The compound 76-1 was obtained by using the same methods as preparing the compound 37-B in embodiment 37, except for the corresponding starting material I2-D. MS m/z: 346.1 [M+H].sup.+

Step 2: Synthesis of Compound 76-2

(511) The compound 76-2 was obtained by using the same methods as preparing the compound 22 in embodiment 22, except for the corresponding starting material. MS m/z: 655.3 [M+H].sup.+

Step 3: Synthesis of Compound 76

(512) The compound 76 was obtained by using the same methods as preparing the compound 2 in embodiment 2, except for the corresponding starting material. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.43-1.58 (m, 4H) 1.61 (s, 6H) 1.69-1.72 (m, 4H) 2.88 (br s, 4H) 3.16-3.20 (m, 4H) 4.76 (br d, J=6.02 Hz, 2H) 4.96 (d, J=17.06 Hz, 1H) 5.06 (d, J=10.04 Hz, 1H) 5.72 (ddt, J=16.94, 10.54, 6.08, 6.08 Hz, 1H) 6.95 (d, J=9.04 Hz, 2H) 7.36 (d, J=7.54 Hz, 1H) 7.47 (br d, J=8.54 Hz, 2H) 7.78 (d, J=8.04 Hz, 1H) 7.85-7.90 (m, 1H) 8.85 (s, 1H). MS m/z: 555.1[M+H].sup.+

Embodiment 76: Synthesis of Compound 77

(513) ##STR00288##

(514) Synthetic Route:

(515) ##STR00289##

Step 1: Synthesis of Compound 77-A

(516) The compound 77-1 was obtained by using the same methods as preparing the compound 22 in embodiment 22, except for the corresponding starting material. MS m/z: 671.3 [M+H].sup.+

Step 2: Synthesis of Compound 77

(517) The compound 77 was obtained by using the same methods as preparing the compound 2 in embodiment 2, except for the corresponding starting material. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.54 (br s, 4H) 1.68-1.75 (m, 4H) 2.89 (br s, 4H) 3.15-3.25 (m, 4H) 4.90-4.95 (m, 2H) 4.96-5.15 (m, 5H) 5.17 (d, J=7.54 Hz, 1H) 5.73 (ddt, J=16.82, 10.42, 6.22, 6.22 Hz, 1H) 6.96 (d, J=8.54 Hz, 2H) 7.38-7.51 (m, 3H) 7.87-7.95 (m, 1H) 8.01 (d, J=8.04 Hz, 1H) 8.86 (s, 1H). MS m/z: 571.1[M+H].sup.+

Embodiment 77: Synthesis of Compound 78

(518) ##STR00290##

(519) Synthetic Route:

(520) ##STR00291##

Step 1: Synthesis of Compound 78-A

(521) Difluorobromomethyl trimethylsilane (761.50 mg, 3.75 mmol), triphenylphosphine (786.73 mg, 3.00 mmol) and 1,3-dimethyl-tetrahydro-2-pyrimidinone (640.75 mg, 5.00 mmol) was added sequentially into 2-bromo-6-acetyl pyridine (500 mg, 2.50 mmol) in acetonitrile (7 mL) solution. The reaction was stirred at 20 C. for 2 hours under nitrogen atmosphere. Potassium hydroxide (3 M, 2.5 mL) solution was added into the reaction mixture, the reaction was stirred at 20 C. for 1.5 hours under nitrogen atmosphere. 2.5 mL 2 mol/L diluted HCl solution was added into the reaction mixture, then stirred for 10 min, saturated sodium bicarbonate solution was added into the reaction mixture, adjusted to pH=7-8, the aqueous phase was extracted by EA (11 mL3), the organic phases were combined and washed by saturated brine (15 mL), dried over anhydrous sodium sulfate, then filtered, the filtrate was evaporated to give the crude product. The crude product was separated by column chromatography (PE/EA=5/1, 3/1) to give 78-1. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.62-1.66 (m, 3H) 4.83 (s, 1H) 5.64-5.95 (m, 1H) 7.50 (dd, J=12.80, 7.78 Hz, 2H) 7.63-7.68 (m, 1H). MS m/z: 253.8 [M+H].sup.+

Step 2: Synthesis of Compound 78-2

(522) The compound 78-2 was obtained by using the same methods as preparing the compound 46-A in embodiment 46, except for the corresponding starting material.

(523) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.68 (s, 3H) 2.61 (s, 3H) 4.66-4.74 (m, 2H) 4.80-4.88 (m, 1H) 4.95 (dd, J=17.08, 1.00 Hz, 1H) 5.08 (d, J=9.54 Hz, 1H) 5.64-6.01 (m, 2H) 7.49 (d, J=7.54 Hz, 1H) 7.91 (d, J=7.54 Hz, 1H) 7.98-8.03 (m, 1H) 8.97 (s, 1H). MS m/z: 394.4 [M+H].sup.+

Step 3: Synthesis of Compound 78

(524) The compound 78 was obtained by using the same methods as preparing the compound 22 in embodiment 22, except for the corresponding starting material.

(525) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.62 (br s, 3H) 1.68 (br s, 8H) 2.33 (s, 3H) 2.43 (br s, 4H) 3.15-3.20 (m, 4H) 4.61-4.69 (m, 1H) 4.73-4.81 (m, 1H) 4.96 (br d, J=17.08 Hz, 1H) 5.07 (d, J=10.28 Hz, 1H) 5.65-6.00 (m, 2H) 6.96 (d, J=9.04 Hz, 2H) 7.41-7.49 (m, 3H) 7.87-7.99 (m, 2H) 8.85 (s, 1H). MS m/z: 605.2 [M+H].sup.+

Embodiment 78: Compound 79

(526) ##STR00292##

(527) Synthetic Route:

(528) ##STR00293## ##STR00294##

Step 1: Synthesis of Compound 79-1

(529) The compound 79-1 was obtained by using the same methods as preparing the compound 22-B in embodiment 22, except for the corresponding starting material. MS m/z: 278.9[M41].sup.+

Step 2: Synthesis of Compound 79-2

(530) The compound 79-2 was obtained by using the same methods as preparing the compound 22-C in embodiment 22, except for the corresponding starting material. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.41 (s, 9H) 3.97-4.07 (m, 4H) 4.26-4.41 (m, 1H) 4.63 (dd, J=16.56, 3.02 Hz, 1H).

Step 3: Synthesis of Compound 79-3

(531) The compound 79-3 was obtained by using the same methods as preparing the compound 22-D in embodiment 22, except for the corresponding starting material. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.31-1.44 (m, 9H) 1.52 (br s, 3H) 2.52 (s, 3H) 4.19-4.64 (m, 6H) 4.75 (dd, J=16.06, 3.01 Hz, 1H) 8.75 (s, 1H). MS m/z: 387.4 [M+H].sup.+

Step 4: Synthesis of Compound 79-4

(532) The compound 79-4 was obtained by using the same methods as preparing the compound 22-E in embodiment 22, except for the corresponding starting material. MS m/z: 287.3 [M+H].sup.+

Step 5: Synthesis of Compound 79-5

(533) The compound 79-5 was obtained by using the same methods as preparing the compound 22-F in embodiment 22, except for the corresponding starting material. MS m/z: 240.9 [M+H].sup.+

Step 6: Synthesis of Compound 79-6

(534) The compound 79-6 was obtained by using the same methods as preparing the compound 22-G in embodiment 22, except for the corresponding starting material. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.59 (s, 6H) 2.59 (s, 3H) 4.09-4.24 (m, 1H) 4.56 (dd, J=15.82, 3.26 Hz, 1H) 4.96 (d, J=15.56 Hz, 2H) 7.42-7.46 (m, 1H) 7.83-7.87 (m, 1H) 7.90-7.95 (m, 1H) 8.94 (s, 1H). MS m/z: 376.0[M+H].sup.+

Step 7: Synthesis of Compound 79

(535) The compound 79 was obtained by using the same methods as preparing the compound 22 in embodiment 22, except for the corresponding starting material. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.61 (s, 6H) 1.62-1.71 (m, 8H) 2.27-2.39 (m, 3H) 2.43 (br s, 4H) 3.14-3.22 (m, 4H) 3.80 (br s, 1H) 4.11-4.25 (m, 1H) 4.57 (dd, J=16.06, 3.52 Hz, 1H) 4.91 (d, J=15.56 Hz, 2H) 6.96 (d, J=9.04 Hz, 2H) 7.34-7.39 (m, 1H) 7.47 (br d, J=8.04 Hz, 2H) 7.85-7.90 (m, 2H) 8.86 (br s, 1H). MS m/z: 587.1 [M+H].sup.+

Embodiment 79: Compound 80

(536) ##STR00295##

(537) Synthetic Route:

(538) ##STR00296##

Step 1: Synthesis of Compound 80-1

(539) At 70-60 C. and under nitrogen atmosphere, n-butyllithium (2.5 M, 5.68 mL) was slowly added dropwise into 2-bromo-5-fluoropyridine (2.5 g, 14.21 mmol) in ether (20 mL) solution, after completion of the addition, the reaction mixture was stirred at 70-60 C. for 0.5 hours, then dry acetone (907.55 mg, 15.63 mmol) was added dropwise into the reaction mixture, the reaction mixture was stirred at 70-60 C. for 1 hour. At 0 C., saturated ammonium chloride (20 mL) solution was added into the reaction mixture to quench the reaction, then the mixture was extracted by EA (20 mL3), the organic phases were combined and washed by saturated brine (20 mL), dried over anhydrous sodium sulfate, then filtered, the filtrate was evaporated to give the crude product, which was purified by silica gel column chromatography (PE/EA=10/1) to give a red compound 80-1. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.51 (d, J=2.02 Hz, 6H) 7.21-7.27 (m, 1H) 7.35 (dd, J=8.54, 3.02 Hz, 1H). MS m/z: 215.9 [M+H].sup.+

Step 2: Synthesis of Compound 80-2

(540) The compound 80-2 was obtained by using the same methods as preparing the compound 22-G in embodiment 22, except for the corresponding starting materials. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.55 (d, J=1.00 Hz, 6H) 2.51 (s, 3H) 4.47 (s, 1H) 4.66 (d, J=6.02 Hz, 2H) 4.87 (dd, J=17.08, 1.00 Hz, 1H) 4.98-5.03 (m, 1H) 5.62 (ddt, J=16.94, 10.42, 6.16, 6.16 Hz, 1H) 7.57 (t, J=9.04 Hz, 1H) 7.71 (dd, J=8.54, 3.01 Hz, 1H) 8.86-8.89 (m, 1H). MS m/z: 376.0 [M+H].sup.+

Step 3: Synthesis of Compound 80

(541) The compound 80 was obtained by using the same methods as preparing the compound 22 in embodiment 22, except for the corresponding starting materials. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.59-1.63 (m, 4H) 1.64 (d, J=1.26 Hz, 6H) 1.66-1.70 (m, 4H) 2.32 (s, 3H) 2.42 (br s, 4H) 3.13-3.22 (m, 4H) 4.65-4.70 (m, 2H) 4.71 (br s, 1H) 4.96 (dd, J=17.08, 1.00 Hz, 1H) 5.07 (d, J=10.28 Hz, 1H) 5.71 (ddt, J=16.82, 10.42, 6.22, 6.22 Hz, 1H) 6.95 (d, J=9.04 Hz, 2H) 7.44 (br d, J=9.04 Hz, 2H) 7.60 (t, J=9.16 Hz, 1H) 7.79 (dd, J=8.66, 2.89 Hz, 1H) 8.84 (s, 1H). MS m/z: 587.1 [M+H].sup.+

Experiment 1: In Vitro Enzymatic Inhibitory Activity of the Compound of the Present Invention

(542) The compounds of the present invention for experimental use were all self-prepared, and their chemical names and structural formulas are shown in the preparation embodiments of the respective compounds. The experimental tests were carried out in Reaction Biology Corporation, USA, and the experimental results were provided by the company.

(543) Experimental Reagents:

(544) Basic reaction buffer: 20 mM hydroxyethylpiperazine ethanesulfuric acid (pH 7.5), 10 mM magnesium chloride, 1 mM EGTA, 0.02% Brij35, 0.02 mg/mL bovine serum albumin, 0.1 mM Na.sub.3VO.sub.4, 2 mM DTT, 1% DMSO

(545) Enzyme: Wee-1 concentration is 150 nM

(546) Matrix: MBP concentration is 20 M

(547) Compound Treatment:

(548) The tested compound was formulated into a 5 mM solution with 100% DMSO, and a 10 points 3-fold gradient dilution was performed with DMSO through an automated pipetting station epMotion 5070.

(549) Experiment Procedure:

(550) 1. Fresh matrix configuration reaction buffer was prepared;

(551) 2. The Wee-1 was added into the matrix solution and shaked gently;

(552) 3. DMSO solution of the compound were added into the kinase reaction mixture using an acoustic technique (Echo 550; nanoliter range) and incubated for 20 minutes at room temperature;

(553) 4. .sup.33P-ATP (specific activity, 10 Ci/L) was added into the reaction mixture to stimulate the reaction;

(554) 5. Incubated for 2 hours at room temperature;

(555) 6. Kinase activity was detected by filter-binding method.

(556) TABLE-US-00002 TABLE 1 In vitro enzymatic activity determination results of the compound of the present invention (IC.sub.50) Compound No. Wee1 (IC.sub.50 nM) 1 67.4 2 43.6 3 24.6 4 250.9 5 90.4 6 101.9 7 82.5 8 51.9 9 108.6 11 48.1 12 25.3 13 42.0 14 47.8 15 31.5 16 52.8 17 42.5 18 131.4 19 83.2 22 238.6 23 79.6 24 48.8 25 20.4 26 14.1 27 24.9 28 13.2 29 13.4 30 58.8 31 15.0 32 64.6 34 120.2 35 240.7 36 57.1 37 328.1 38 20.7 39 620.4 40 30.7 41 13.1 42 17.4 43 12.1 44 12.6 45 7.4 46 44.6 47 3.0 48 6.3 49 3.1 50 2.5 51 2.8 52 71 53 63 54 57 55 90 56 74 57 47 58 73 59 22.6 60 14.8 61 15.7 62 36.1 63 8.3 64 9.9 65 41.6 66 15.0 67 12.8 68 30.4 69 5.7 70 20.3 71 15.7 72 8.2 73 5.3 74 13.0 75 17 76 13 77 9 78 32 79 43 80 49
Experimental Conclusion:

(557) According to Table 1, the compounds of the present invention have a good inhibitory effect on Wee1 kinase.

Experiment 2: Pharmacokinetic Evaluation of the Compound

(558) Experimental objective: pharmacokinetics evaluation of the compound in BALB/c nude mice.

(559) Experimental Material:

(560) BALB/c mice (female)

(561) Experimental Procedure:

(562) The rodent pharmacological characteristics of the compound after intravenous administration and oral administration were tested by a standard protocol, the candidate compound in the experiment was formulated into a clear solution, and the mice were administered by a single intravenous injection and oral administration. The intravenous and oral vehicles are a certain proportion of aqueous hydroxypropyl 3-cyclodextrin or physiological saline solution. Whole blood samples within 24 hours were collected, centrifuged at 3000 g for 15 minutes, and the supernatant was separated to obtain plasma samples, 4 times volume containing internal standard of acetonitrile solution was added to precipitate protein, centrifuged to remove the supernatant and equal volume of water was added and again centrifuged to remove the supernatant, the plasma concentration was quantitatively analyzed by LC-MS/MS analysis, and the pharmacokinetic parameters such as peak concentration, peak time, clearance rate, half-life, area under the curve of the drug, and bioavailability were calculated.

(563) Experimental Results:

(564) TABLE-US-00003 TABLE 2 pharmacokinetic test results Tested compound Clearance Concentration prepared by rate Half-life integral AUC bioavailability embodiments (mL/min/kg) T.sub.1/2 (h) (nM .Math. hr) F (%) AZD1775 85.7 0.25 1200 31.0 Embodiment 57.1 1.7 1729 35.3 15 Embodiment 51 1.49 2151 32.4 29 Embodiment 23 1.81 5593 45.1 43

(565) Conclusion: the compounds of the present invention can significantly improve the pharmacokinetics indexes in mice, reduce the clearance rate of the compound in vivo, increase the half-life time, and significantly increase the concentration integral.

Experiment 3: In Vivo Drug PD Research

(566) (1) In vivo PD study of the compound used on tumors of human colon cancer LoVo cells subcutaneously xenografted to BALB/c nude mice model

(567) Experimental method: the experimental animals used were BALB/c nude mice (provided by Shanghai Xipuer-Beikai Experimental animal CO., Ltd), 6-8 weeks old, weighting 18-22 g.

(568) Human colon cancer LoVo cells, monolayer-cultured in vitro, culturing condition was Ham's F-12 medium supplemented with 10% fetal bovine serum, 100 U/mL penicillin, 100 g/mL streptomycin and 2 mM glutamine, 37 C., 5% CO.sub.2. Conventional digestion treatment passage was done with trypsin-EDTA twice a week. When the cell saturation reached 80%-90%, the cells were collected, counted, and inoculated. 0.1 mL (10106) of LoVo cells were subcutaneously inoculated into the right back of each nude mouse, and group administration was started when the average tumor volume reached 213 mm.sup.3. The administration mode was oral administration, 40 mg/kg twice a day for three consecutive weeks.

(569) After the last administration of the experiment, a portion of plasma, tumor, intestine, brain, lung, liver and pancreas was collected from the number 1-2 animals after 0.5 h of administration, and quick-froze for PK detection; a portion of plasma, tumor, intestine, brain, lung, liver and pancreas was collected from the number 3-4 animals after 2 h of administration, and quick-froze for PK detection; a portion of plasma, tumor, intestine, brain, lung, liver and pancreas was collected from the number 5-6 animals after 8 h of administration, and quick-froze for PK detection. The tumor, intestine, brain, lung, liver and pancreas were weighted separately and added 9 times volumes of homogenate MeOH/15 mM PBS (1:2, v:v) for homogenization. 20 L sample was added into 300 L of the stop buffer including internal standard for protein precipitation, and shaked to mix well. The mixture was centrifuged at 13,000 rpm for 10 minutes at 20-25 C., and 3 L of the supernatant was taken for LCMS analysis.

(570) The final experimental results was shown in Table 3:

(571) TABLE-US-00004 TABLE 3 distribution result for the mice tissues Body part Parameter AZD1775 Embodiment 30 plasma Cmax (nM) 3070 4270 Tmax (h) 0.5 2 AUC.sub.0-last (nM .Math. h) 6970 14183 Brain Cmax (nM) ND 959 Tmax (h) ND 0.5 AUC.sub.0-last (nM .Math. h) ND 2124 Colon Cmax (nM) 238000 134950 Tmax (h) 0.5 0.5 AUC.sub.0-last (nM .Math. h) 225458 401622 liver Cmax (nM) 38100 67700 Tmax (h) 0.5 2 AUC.sub.0-last (nM .Math. h) 117325 256862 lung Cmax (nM) 17240 14600 Tmax (h) 0.5 2 AUC.sub.0-last (nM .Math. h) 36531 74704 pancreas Cmax (nM) 35650 49300 Tmax (h) 0.5 2 AUC.sub.0-last (nM .Math. h) 55530 253458 tumer Cmax (nM) 3855 5425 Tmax (h) 2 8 AUC.sub.0-last (nM .Math. h) 15291 37263

(572) Conclusion: The compound of the present invention may significantly enhance its exposed quantity in mice tissues.

Biological Activity Experiment 4: Inhibition Experiment for hERG Potassium Channel

(573) Experimental objective: effect detection on the tested compounds on hERG potassium channel with automatic patch clamp method.

(574) Experimental Procedure

(575) 1. Preparation of the Compounds

(576) On the day of the experiment, a 20 mM compound mother solution was serially diluted 3 times with 100% DMSO, i.e., 10 L of a 20 mM compound mother solution was added to 20 L of DMSO, and intermediate concentrations of 6 consecutively diluted DMSO compounds were sequentially obtained. Then 10 L of the intermediate concentration of the compound was added to 4990 L of the extracellular solution, and the final concentration to be tested was obtained by 500-fold dilution, with a highest test concentration of 40 M, where the concentrations were respectively 40, 13.3, 4.4, 1.48, 0.49, and 0.16 M. Positive control compound cisapride preparation: 150 M cisapride mother liquor was serially diluted 3 times with 100% DMSO, i.e., 10 L of 150 M cisapride mother liquor was added to 20 L of DMSO, and 5 intermediate concentrations of cisapride serially diluted with DMSO were sequentially obtained. Then 10 L of cisapride intermediate concentration was added to 4990 L of extracellular fluid, the final concentration to be tested was obtained by a 500-fold dilution, with a highest test concentration of 300 nM, where the 5 concentrations was 300, 100, 33.3, 11.1 and 3.70 nM, respectively. The DMSO content in the final test concentration did not exceed 0.2%, and this concentration of DMSO had no effect on the hERG potassium channel.

(577) 2. Electrophysiological Recording Process

(578) CHO (Chinese Hamster Ovary) cells that stably expressed hERG potassium channels were recorded for hERG potassium channel currents using whole cell patch clamp technique at room temperature. A glass microelectrode with the tip resistance of about 2-5 M is connected to an Axopatch 200B (Molecular Devices) patch clamp amplifier. The clamping voltage and data recording were controlled and recorded by the pClamp 10 software via computer control with a sampling frequency of 10 kHz and a filtering frequency of 2 kHz. After obtaining the whole cell recording, the cells were clamped at 80 mV, and the step voltage of the induced hERG potassium current (I hERG) was given a 2 s depolarization voltage from 80 mV to +20 mV, and then repolarized to 50 mV for 1 s, and finally returned to 80 mV. This voltage stimulation was given every 10 s, and the administration process was started after confirmation of the stabilization of the hERG potassium current (1 minute). Compound concentrations were administered continuously starting from a low test concentration and each test concentration was given for 1 minute. Each concentration was tested with at least 3 cells (n3).

(579) 3. Data Processing

(580) Data analysis and processing were performed by pClamp 10, Patch Master, GraphPad Prism 5 and Excel. Inhibition degrees of different compound concentrations on the hERG potassium current (tail current peaks of hERG induced at 50 mV) was calculated by the formula: Fractional block %=[1(I/Io)]100%, wherein, Fractional block represents for the inhibition percentage of the potassium current, I and Io represent for the hERG potassium current amplitude before and after dosing. IC.sub.50 of the compound was fitting calculated by the formula I/Io=1/{1+([C]/IC50){circumflex over ()}n},

(581) wherein, Io and I represents for hERG potassium current amplitudes before and after dosing respectively. [C] was the concentration of the compound, n was Hill index.

(582) 4. Solution

(583) Extracellular fluid formula (mM): 140 NaCl, 5 KCl, 1 CaCl.sub.2, 1.25 MgCl.sub.2, 10 HEPES and 10 Glucoses, pH was adjusted to 7.4 by NaOH. Intracellular fluid formula (mM): 140 KCl, 1 MgCl.sub.2, 1 CaCl.sub.2, 10 EGTA and 10 HEPES, the pH was adjusted to 7.2 by KOH.

Abbreviation

(584) HEPES: 4-(2-hydroxyethyl)piperazin-1-ethanesulfonic acid, N-(2-hydroxyethyl)piperazin-N-(2-ethanesulfonic acid)

(585) EGTA: ethylene glycol bis(2-aminoethyl ether) tetraacetic acid

(586) 5. Quality Control

(587) the experimental data in the report satisfied the quality control standard:

(588) Whole cell sealing impedance >1 G

(589) Series resistance compensation would be more than 80% if the resistance was greater than 10 M

(590) hERG tail current amplitude >400 pA

(591) attenuation <2% per min

(592) current stability: 6 recorded tail current peaks and averages would not exceed 2%

(593) Pharmacological indicators: multi-concentration cisapride inhibition effect on hERG channel was used as positive control

(594) 6. Experimental Result

(595) hERG IC.sub.50 values of the compounds of the embodiments was shown in Table 4.

(596) TABLE-US-00005 TABLE 4 hERG IC.sub.50 value result for the compound of the embodiments Tested sample hERG IC.sub.50 (nM) Test times AZD1775 11.82 N = 4 Embodiment 17 >30 N = 2 Embodiment 29 29.03 N = 2

(597) Conclusion: the compound of the present invention may significantly decrease activity of hERG thus improve safety.

Experiment 5: Thermodynamic Solubility Determination

(598) Method:

(599) 1. Preparation, Shaking and Filtration of the Sample

(600) No less than 2 mg sample powder was weighted in a Whatman miniuniprep vial. If the experiment required to test the thermodynamic solubility of the sample in multiple buffer solutions, then separate vial was required for each test.

(601) 450 L buffer (pH=7.4) was added into each of the Whatman miniuniprep vial. After addition of the buffer, the Whatman filter cap of the miniuniprep is mounted and pressed until it was above the liquid level to allow the filter to contact with the buffer solution during shaking. The solubility sample was vortexed for 2 minutes. And the solution phenomenon was recorded. Then the vial was shaked at room temperature (about 22 to 25 C.) for 24 hours at a speed of 550 rpm. And the Whatman Miniunipreps filter cap was pressed to bottom to obtain the filtrate of the solubility solution of the sample. All sample vials should conduct filtration for the insoluble matter before and after and osmotic phenomenon. The buffer was diluted 50 times to obtain a sample dilution.

(602) 2 Analysis and Detection

(603) Three UV standard solutions were injected from low to high concentrations into HPLC, and then dilution and supernatant of the test compounds were injected. The sample was injected and tested twice. Integration was conducted to the UV peaks. The standard curve was established and the thermodynamic solubility of the sample was calculated.

(604) TABLE-US-00006 TABLE 5 Compound Solubility (M) pH 6.5 Solubility (M) pH 7.4 AZD1775 434.16 68.77 Embodiment 29 1467.99 1046.27

(605) As shown in Table 5, comparing to the AZD1775, the embodiment 29 of the present invention shows excellent solubility in water (under pH=6.5 and pH=7.4). Therefore, it's obvious that the compound of the present invention is easier to be dissolved in water.