Substituted-indole-compounds as estrogen receptor down-regulators
10519143 ยท 2019-12-31
Assignee
Inventors
- Jianyu Lu (Shanghai, CN)
- Charles Z. Ding (Shanghai, CN)
- Lihong HU (Shanghai, CN)
- Huijun He (Shanghai, CN)
- Shuhui Chen (Shanghai, CN)
- Jiaqiang DONG (Shanghai, CN)
- Tie-Lin Wang (Shanghai, CN)
Cpc classification
C07D409/06
CHEMISTRY; METALLURGY
A61P35/00
HUMAN NECESSITIES
C07D209/18
CHEMISTRY; METALLURGY
C07D405/06
CHEMISTRY; METALLURGY
C07D403/12
CHEMISTRY; METALLURGY
A61P15/00
HUMAN NECESSITIES
C07D401/06
CHEMISTRY; METALLURGY
International classification
A61K31/437
HUMAN NECESSITIES
C07D409/06
CHEMISTRY; METALLURGY
C07D403/12
CHEMISTRY; METALLURGY
C07D209/18
CHEMISTRY; METALLURGY
C07D401/06
CHEMISTRY; METALLURGY
Abstract
Disclosed is a new indole compound, in particular, the compound as shown in formula (I), and a preparation method, pharmaceutical composition and use thereof as an estrogen receptor down-regulator in preparing drugs for treating estrogen receptor-positive breast cancer. ##STR00001##
Claims
1. A compound represented by formula (I), a pharmaceutically acceptable salt or a hydrate thereof, ##STR00155## wherein, R.sub.1 is selected from ##STR00156## X is selected from single bond, O and S; Y and Z are CH; ring A is selected from 5-10 membered aryl and 5-10 membered heteroaryl; R.sub.2 is selected from H, halogen, CN, NO.sub.2, OH, COOH, NH.sub.2, or the group consisting of C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, C.sub.3-6 cycloalkyl and 3-6 membered heterocycloalkyl, each of which is optionally substituted with 1, 2, or 3 R; R.sub.3 is selected from H, halogen, CN, NO.sub.2, OH, COOH, NH.sub.2, or the group consisting of C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, C.sub.3-6 cycloalkyl and 3-6 membered heterocycloalkyl, each of which is optionally substituted with 1, 2, or 3 R; R.sub.4 and R.sub.5 are each independently selected from H, halogen, CN, NO.sub.2, OH, COOH, NH.sub.2, or the group consisting of C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, C.sub.3-6 cycloalkyl and 3-6 membered heterocycloalkyl, each of which is optionally substituted with 1, 2, or 3 R; R.sub.6 is selected from the group consisting of C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, C.sub.3-6 cycloalkyl and 3-6 membered heterocycloalkyl, each of which is optionally substituted with 1, 2, or 3 R; R is selected from H, F, Cl, Br, I, OH, CN, NH.sub.2, COOH, C(O)NH.sub.2, or the group consisting of C.sub.1-8 alkyl, C.sub.1-8 heteroalkyl, C.sub.3-6 cycloalkyl, 3-6 membered heterocycloalkyl and C.sub.3-6 cycloalkyl-C.sub.1-3 alkyl-, each of which is optionally substituted with 1, 2, or 3 R; n is selected from 0, 1, 2, 3 and 4; m is selected from 0, 1, 2, 3 and 4; P is selected from 0, 1, 2 and 3; or, when m is 2, R.sub.3 and R.sub.3 are connected together to form a 5-6 membered ring; R is selected from the group consisting of F, Cl, Br, I, OH, CN, NH.sub.2, COOH, Me, Et, CF.sub.3, CHF.sub.2, CH.sub.2F, NHCH.sub.3 and N(CH.sub.3).sub.2; hetero represents a heteroatom or a heteroatom group, which is selected from the group consisting of C(O)N(R), N(R), S(O).sub.2N(R), S(O) N(R), O, S, O, S, ON, C(O)O, C(O), C(S), S(O), S(O).sub.2 and N(R)C(O)N(R); in any of the above cases, the number of the heteroatom or the heteroatom group is independently selected from 1, 2 and 3.
2. A compound represented by formula (I), a pharmaceutically acceptable salt or a hydrate thereof, ##STR00157## wherein, R.sub.1 is selected from ##STR00158## Y and Z are CH; ring A is selected from 5-10 membered aryl and 5-10 membered heteroaryl; R.sub.2 is selected from H, halogen, CN, NO.sub.2, OH, COOH, NH.sub.2, or the group consisting of C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, C.sub.3-6 cycloalkyl and 3-6 membered heterocycloalkyl, each of which is optionally substituted with 1, 2, or 3 R; R.sub.3 is selected from H, halogen, CN, NO.sub.2, OH, COOH, NH.sub.2, or the group consisting of C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, C.sub.3-6 cycloalkyl and 3-6 membered heterocycloalkyl, each of which is optionally substituted with 1, 2, or 3 R; R.sub.4 and R.sub.5 are each independently selected from H, halogen, CN, NO.sub.2, OH, COOH, NH.sub.2, or the group consisting of C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, C.sub.3-6 cycloalkyl and 3-6 membered heterocycloalkyl, each of which is optionally substituted with 1, 2, or 3 R; R.sub.6 is selected from the group consisting of C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, C.sub.3-6 cycloalkyl and 3-6 membered heterocycloalkyl, each of which is optionally substituted with 1, 2, or 3 R; R is selected from H, F, Cl, Br, I, OH, CN, NH.sub.2, COOH, C(O)NH.sub.2, or the group consisting of C.sub.1-8 alkyl, C.sub.1-8 heteroalkyl, C.sub.3-6 cycloalkyl, 3-6 membered heterocycloalkyl and C.sub.3-6 cycloalkyl-C.sub.1-3 alkyl-, each of which is optionally substituted with 1, 2, or 3 R; n is selected from 0, 1, 2, 3 and 4; m is selected from 0, 1, 2, 3 and 4; or, when m is 2, R.sub.3 and R.sub.3 are connected together to form a 5-6 membered ring; R is selected from the group consisting of F, Cl, Br, I, OH, CN, NH.sub.2, COOH, Me, Et, CF.sub.3, CHF.sub.2, CH.sub.2F, NHCH.sub.3 and N(CH.sub.3).sub.2; hetero represents a heteroatom or a heteroatom group, which is selected from the group consisting of C(O)N(R), N(R), S(O).sub.2N(R), S(O)N(R), O, S, O, S, ON, C(O)O, C(O), C(S), S(O), S(O).sub.2 and N(R)C(O)N(R); in any of the above cases, the number of the heteroatom or the heteroatom group is independently selected from 1, 2 and 3.
3. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 2, wherein, R is selected from H, F, Cl, Br, I, OH, CN, NH.sub.2, COOH, or the group consisting of C.sub.1-5 alkyl, C.sub.1-5 heteroalkyl, C.sub.3-6 cycloalkyl-C.sub.1-3 alkyl-, each of which is optionally substituted with 1, 2, or 3 R.
4. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 3, wherein, R is selected from H, F, Cl, Br, I, OH, CN, NH.sub.2, COOH, C(O)NH.sub.2, or the group consisting of CH.sub.3, CH.sub.3CH.sub.2, S(O)CH.sub.3, ##STR00159## each of which is optionally substituted with 1, 2, or 3 R.
5. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 4, wherein, R is selected from the group consisting of H, F, Cl, Br, I, OH, CN, NH.sub.2, COOH, C(O)NH.sub.2, CH.sub.3, CH.sub.2Cl, CH.sub.2F, CHF.sub.2, CF.sub.3, OCF.sub.3, CH.sub.2OH, Et, S(O)CH.sub.3, ##STR00160##
6. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 1, wherein, R is selected from H, F, Cl, Br, I, OH, CN, NH.sub.2, COOH, or the group consisting of C.sub.1-5 alkyl, C.sub.1-5 heteroalkyl and C.sub.3-6 cycloalkyl-C.sub.1-3 alkyl-, each of which is optionally substituted with 1, 2, or 3 R.
7. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 6, wherein, R is selected from H, F, Cl, Br, I, OH, CN, NH.sub.2, COOH, C(O)NH.sub.2, or the group consisting of CH.sub.3, CH.sub.3CH.sub.2, S(O)CH.sub.3, ##STR00161## each of which is optionally substituted with 1, 2, or 3 R.
8. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 7, wherein, R is selected from the group consisting of H, F, Cl, Br, I, OH, CN, NH.sub.2, COOH, C(O)NH.sub.2, CH.sub.3, CH.sub.2Cl, CH.sub.2F, CHF.sub.2, CF.sub.3, OCF.sub.3, CH.sub.2OH, Et, S(O)CH.sub.3, ##STR00162##
9. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 1, wherein, the structural unit ##STR00163## is selected from ##STR00164##
10. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 9, wherein, the structural unit ##STR00165## is selected from ##STR00166##
11. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 10, wherein, the structural unit ##STR00167## is selected from the group consisting of ##STR00168##
12. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 1, wherein, R.sub.2 is selected from H, F, Cl, Br, I, CN, NO.sub.2, OH, COOH, NH.sub.2, or the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, C.sub.1-4 alkylthio, NH(C.sub.1-4 alkyl), N,N-di(C.sub.1-3 alkyl)amino and C.sub.3-6 cycloalkyl, each of which is optionally substituted with 1, 2, or 3 R, and/or, R.sub.3 is selected from H, F, Cl, Br, I, CN, NO.sub.2, OH, COOH, NH.sub.2, or the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, C.sub.1-4 alkylthio, NH(C.sub.1-4 alkyl), N,N-di(C.sub.1-3 alkyl)amino, C.sub.3-6 cycloalkyl, 3-6 membered heterocycloalkyl, 5-6 membered aryl and 5-6 membered heteroaryl, each of which is optionally substituted with 1, 2, or 3 R, and/or, R.sub.4 and R.sub.5 are each independently selected from H, F, Cl, Br, I, CN, NO.sub.2, OH, COOH, NH.sub.2, or the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, C.sub.1-4 alkylthio, NH(C.sub.1-4 alkyl), N,N-di(C.sub.1-3 alkyl)amino and C.sub.3-6 cycloalkyl, each of which is optionally substituted with 1, 2, or 3 R, and/or, ring A is selected from the group consisting of phenyl, pyridyl, pyrazinyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, thienyl, benzofuranyl, benzothienyl, indolyl, benzimidazolyl, benzothiazolyl, purinyl, quinolinyl and isoquinolinyl, and/or, R.sub.6 is selected from ##STR00169##
13. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 12, wherein, R.sub.3 is selected from H, F, Cl, Br, I, CN, NO.sub.2, OH, COOH, NH.sub.2, or the group consisting of CH.sub.3, CH.sub.3CH.sub.2, CH.sub.3O, CH.sub.3CH.sub.2O and ##STR00170## each of which is optionally substituted with 1, 2, or 3 R.
14. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 13, wherein, R.sub.3 is selected from the group consisting of H, F, Cl, CN, CH.sub.3, CF.sub.3 and ##STR00171##
15. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 12, wherein, R.sub.4 and R.sub.5 are each independently selected from the group consisting of H, F, Cl, Br, CN and CH.sub.3.
16. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 12, wherein, the structural unit ##STR00172## is selected from the group consisting of ##STR00173##
17. The compound, the pharmaceutically acceptable salt the hydrate thereof as defined in claim 16, wherein, the structural unit ##STR00174## is selected from the group consisting of ##STR00175##
18. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 2, wherein, the structural unit ##STR00176## is selected from ##STR00177##
19. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 18, wherein, the structural unit ##STR00178## is selected from ##STR00179##
20. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 1, wherein, the compound is selected from the group consisting of ##STR00180## ##STR00181## wherein, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are defined as claim 1.
21. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 2, wherein, the compound is selected from ##STR00182## wherein, R.sub.3, R.sub.5 and R are defined as in claim 2.
22. The compound, pharmaceutically acceptable salt or hydrate thereof as defined in claim 1, wherein, the compound is selected from the group consisting of ##STR00183## ##STR00184## ##STR00185## ##STR00186## ##STR00187## ##STR00188## ##STR00189## ##STR00190## ##STR00191## ##STR00192## ##STR00193## ##STR00194## ##STR00195## ##STR00196## ##STR00197## ##STR00198##
23. A pharmaceutical composition comprising a therapeutically effective amount of the compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 1 and a pharmaceutically acceptable carrier.
24. A method for treating a disorder associated with estrogen receptor in a subject in need thereof, comprising: administrating an effective amount of the compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 1, wherein, the disorder associated with estrogen receptor is estrogen receptor-positive breast cancer.
25. The compound, the pharmaceutically acceptable salt or the hydrate thereof as defined in claim 2, wherein, the compound is selected from the group consisting of ##STR00199##
Description
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT
(1) The present invention will be specifically described below by way of embodiments, but the scope of the present invention is not limited thereto.
(2) It will be appreciated by the skilled in the art that the order of the reaction step in any reaction scheme may be varied in order to prepare the compound of the present invention, which is also encompassed within the scope of the invention.
Embodiment 1
(3) ##STR00069## ##STR00070## ##STR00071##
(4) Step A: N-butyllithium (2.5M, 428.40 mL, 1.05 eq) was slowly added dropwise to a solution of 1 (100.00 g, 1.02 mol, 140.85 mL, 1.00 eq) in tetrahydrofuran (500 mL) over 1 hour at 75 C. under nitrogen atmosphere. The reaction solution was allowed to warm to 0 C. and stirred for 10 minutes, then cooled to 75 C., and hexamethylphosphoric triamide (201.06 g, 1.12 mol, 197.12 mL, 1.10 eq) was added over 1 hour. After the reaction solution was stirred at 75 C. for 1 hour, ethyl iodide (198.86 g, 1.27 mol, 101.98 mL, 1.25 eq) was added over 1 hour. The reaction solution was allowed to warm to 20 C. and stirred for 10 hours. Then 400 mL water was added, and the mixture was partitioned. The organic phase was washed three times with 400 mL water, dried over anhydrous sodium sulfate, filtered and purified by distillation to give the product 2 as a colorless liquid (65.00 g, 514.77 mmol, yield 50.47%).
(5) Step B: Triethyl phosphonoacetate 4 (11.60 g, 51.72 mmol, 10.26 mL, 1.20 eq) and lithium chloride (3.65 g, 86.20 mmol, 1.77 mL, 2.00 eq) were added to a solution of 3 (10.00 g, 43.10 mmol, 1.00 eq) in 100 mL acetonitrile, then a solution of DBU (8.53 g, 56.03 mmol, 8.45 mL, 1.30 eq) in acetonitrile was added dropwise over 30 min at 0 C. under nitrogen atmosphere. After the reaction solution was stirred at 15 C. for 1 hour, 100 mL water was added, and the mixture was partitioned. The aqueous phase was extracted twice with 70 mL dichloromethane. The organic phase was combined, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by silica gel column chromatography (PE:EA=100:1-30:1) to give the product 5 as a yellow solid (12.00 g, 39.72 mmol, yield 92.16%).
(6) Step C: Dimethylaminopyridine (3.65 g, 29.88 mmol, 0.10 eq) and Boc.sub.2O (68.46 g, 313.70 mmol, 72.07 mL, 1.05 eq) were added to a solution of 6 (35.00 g, 298.76 mmol, 1.00 eq) in 400 mL dichloromethane. The reaction solution was stirred at 20 C. for 12 hours, then washed twice with 400 mL aqueous ammonium chloride solution. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give the product 7 as a colorless oil (60.00 g, 276.17 mmol, yield 92.44%).
(7) Step D: Lithium diisopropylamide (2M, 75.95 mL, 1.10 eq) was slowly added dropwise to a solution of 7 (30.00 g, 138.08 mmol, 1.00 eq) in tetrahydrofuran (400 mL) at 75 C. under nitrogen atmosphere. The reaction solution was stirred at 75 C. for 30 minutes, then cyanogen bromide (55.40 g, 523.04 mmol, 38.47 mL, 3.79 eq) was added. The reaction solution was allowed to warm to 15 C. and stirred for 12 hours, then 400 mL water was added. After the mixture was partitioned, the organic phase was washed three times with 300 mL water, dried over anhydrous sodium sulfate and filtered to give a crude product, which was purified by silica gel column chromatography (PE:EA=1:0-50:1) to give the product 8 as a yellow oil (39.00 g, crude product).
(8) Step E: Cesium carbonate (85.81 g, 263.38 mmol, 2.00 eq), cuprous iodide (1.25 g, 6.58 mmol, 0.05 eq), palladium acetate (1.48 g, 6.58 mmol, 0.05 eq) and 1,1-bis(diphenylphosphino)ferrocene (3.65 g, 6.58 mmol, 0.05 eq) were added to a solution of 8 (39.00 g, 131.69 mmol, 1.00 eq) in 300 mL N,N-dimethylacetamide, followed by addition of 2 (33.26 g, 263.38 mmol, 2.00 eq) under nitrogen atmosphere. The reaction solution was stirred at 80 C. for 12 hours, then 1 L ethyl acetate and 1 L water were added. After the mixture was filtered and partitioned, the organic phase was washed three times with 1 L water, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by silica gel column chromatography (PE:EA=1:0-30:1) to give the product 9 as a yellow oil (27.00 g, crude product).
(9) Step F: Potassium carbonate (69.27 g, 501.25 mmol, 5.00 eq) was added to a solution of 9 (27.00 g, 100.25 mmol, 1.00 eq) in 300 mL methanol and 15 mL water. The reaction solution was stirred at 70 C. for 12 hours, then filtered and concentrated. 300 mL ethyl acetate was added and washed twice with 300 mL water. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by silica gel column chromatography (PE:EA=100:1-30:1) to give the product 10 as a yellow solid (7.50 g, 44.32 mmol, yield 44.21%). MS [ESI, M+1]: 170.1.
(10) Step G: diboron pinacol ester (4.50 g, 17.73 mmol, 1.00 eq) and tetrakis(triphenylphosphine)platinum (1.10 g, 886.50 umol, 0.05 eq) were added to a solution of 5 (3.00 g, 17.73 mmol, 1.00 eq) in 30 mL dimethyltetrahydrofuran. The reaction solution was stirred at 70 C. under nitrogen atmosphere for 5 hours, and then cooled to room temperature. The product 6 contained in the reaction solution was used in the next step without purification.
(11) Step H: Cesium carbonate (11.55 g, 35.44 mmol, 2.00 eq), compound 5 (4.28 g, 14.18 mmol, 0.80 eq) and bis(triphenylphosphine)palladium dichloride (622.02 mg, 886.00 umol, 0.05 eq) were added to a solution of 10 (7.50 g, 17.72 mmol, 1.00 eq) in 70 mL dimethyltetrahydrofuran and 3 mL water. The reaction solution was stirred at 15 C. under nitrogen atmosphere for 12 hours. The product 11 contained in the reaction solution was used in the next step without purification.
(12) Step I: 2-Chloro-4-fluoroiodobenzene (9.03 g, 35.22 mmol, 2.00 eq), potassium hydroxide solution (4M, 22.01 mL, 5.00 eq) and bis(triphenylphosphine)palladium dichloride (617.94 mg, 880.50 umol, 0.05 eq) were added to a solution of 11 (8.30 g, 17.61 mmol, 1.00 eq) in 100 mL dimethyltetrahydrofuran. The reaction solution was stirred at 70 C. under nitrogen atmosphere for 12 hours, then filtered through celite, and the filtrate was washed twice with 100 mL brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by silica gel column chromatography (PE:EA=40: 1-10:1) to give the product 13 as a yellow solid (4.50 g, 5.57 mmol, yield 31.65%, purity 58.7%). MS [ESI, M+1]: 474.3.
(13) Step J: Lithium hydroxide (1.33 g, 55.73 mmol, 10.00 eq) was added to a solution of 13 (4.50 g, 5.57 mmol, 1.00 eq) in a mixed solvent of 30 mL methanol, 30 mL tetrahydrofuran and 10 mL water. The reaction solution was stirred at 35 C. for 1 hour, then 30 mL water was added. The reaction solution was adjusted to pH 5 with 1M hydrochloric acid, then extracted twice with 50 mL ethyl acetate. The organic phase was combined, washed twice with 60 mL water, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative HPLC (formic acid system) to give the product I-1 (2.10 g, 4.69 mmol, yield 84.13%, purity 99.5%). MS [ESI, M+1]: 446.1.
(14) .sup.1H NMR EW3644-175-P1B (400 MHz, DMSO-d.sub.6): 12.34 (s, 1H), 10.75 (s, 1H), 7.56 (d, J=7.6 Hz, 1H), 7.47-7.42 (m, 3H), 7.36 (dd, J=8.8 Hz, J=1.6 Hz, 1H), 7.30 (d, J=8.0 Hz, 1H), 7.25 (dd, J=8.8 Hz, J=6.4 Hz, 1H), 7.12 (dt, J=2.4 Hz, J=8.4 Hz, 1H), 7.06 (t, J=7.6 Hz, 1H), 7.01-6.96 (m, 3H), 6.57 (s, 1H), 6.41 (d, J=16.0 Hz, 1 H), 2.71-2.59 (m, 2 H), 0.99 (t, J=7.2 Hz, 3 H).
Embodiment 2
(15) ##STR00072##
(16) Step A: A solution of compound 12 (556.08 mg, 1.18 mmol, 1.00 eq), 3-trifluoromethoxybromobenzene (572.85 mg, 2.11 mmol, 1.80 eq), aqueous potassium hydroxide solution (4M, 1.64 mL, 5.60 eq), dichlorobis(triphenylphosphine)palladium (24.64 mg, 35.10 umol, 0.03 eq) in 20 mL 2-methyltetrahydrofuran was purged with nitrogen for three times. The reaction solution was stirred at 80 C. under nitrogen atmosphere for 24 hours, then diluted with 20 mL water, extracted twice with 50 mL ethyl acetate each time. The organic phase was combined, washed once with 50 mL saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to give the compound 14 as a yellow oil (200.00 mg), which was directly used in the next step.
(17) Step B: Lithium hydroxide monohydrate (83.00 mg, 1.98 mmol, 5.00 eq) was added to a solution of compound 14 (200.00 mg, 408.56 umol, 1.00 eq) in 20 mL methanol and 6 mL water, the reaction solution was stirred at 30 C. for 3 hours, then poured into 100 mL water. The mixture was adjusted to pH 5-6 with 3 mol/L hydrochloric acid, then extracted twice with 100 mL dichloromethane each time. The organic phase was combined, dried over anhydrous sodium sulfate, filtrated and concentrated to give a crude product, which was purified by preparative chromatography (formic acid) to give I-2 (22.00 mg, 46.08 umol, yield 11.65%). MS [ESI, M+1]: 478.1.
(18) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.78 (br. s., 1H), 7.55 (d, J=7.2 Hz, 1H), 7.50-7.27 (m, 5H), 7.23 (d, J=7.6 Hz, 1H), 7.14 (d, J=7.6 Hz, 1H), 7.10-6.85 (m, 5H), 6.54 (br. s., 1H), 6.42 (d, J=16.0 Hz, 1H), 2.65-2.78 (m, 2H), 0.98 (t, J=6.8 Hz, 3H).
Embodiment 3
(19) ##STR00073##
(20) Step A: A solution of compound 12 (556.08 mg, 1.18 mmol, 1.00 eq), 2-trifluoromethoxybromobenzene (281.98 mg, 1.17 mmol, 174.06 uL, 1.00 eq), aqueous potassium hydroxide solution (4M, 1.64 mL, 5.60 eq) and dichlorobis(triphenylphosphine)palladium (24.64 mg, 35.10 umol, 0.03 eq) in 20 mL 2-methyltetrahydrofuran was purged with nitrogen for three times. The reaction solution was stirred at 80 C. under nitrogen atmosphere for 24 hours, then diluted with 20 mL water. The mixture was extracted twice with 50 mL ethyl acetate each time. The organic phase was combined, washed once with 100 mL saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to give the compound 15 as a yellow oil (380.00 mg), which was directly used in the next step.
(21) Step B: Lithium hydroxide monohydrate (83.00 mg, 1.98 mmol, 5.00 eq) was added to a solution of compound 15 (380.00 mg, 751.69 umol, 1.00 eq) in a mixed solvent of 6 mL tetrahydrofuran, 20 mL methanol and 6 mL water, the reaction solution was stirred at 30 C. for 7 hours. The reaction solution was concentrated to 1 mL, then adjusted to pH 5-6 with 3 mol/L hydrochloric acid, extracted twice with 50 mL dichloromethane each time. The organic phase was combined, dried over anhydrous sodium sulfate, filtrated and concentrated to give a crude product, which was purified by preparative chromatography (formic acid) to give I-3 (45.00 mg, 94.25 umol, yield 12.54%). MS [ESI, M+1]: 478.0.
(22) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.77 (s, 1H), 7.57 (d, J=7.6 Hz, 1H), 7.50-7.37 (m, 3H), 7.36-7.15 (m, 5H), 7.11-6.97 (m, 2H), 6.92 (d, J=8.4 Hz, 2H), 6.57 (s, 1H), 6.42 (d, J=16.4 Hz, 1H), 2.80-2.60 (m, 2H), 0.97 (t, J=7.6 Hz, 3H).
Embodiment 4
(23) ##STR00074##
(24) Step A: 3-Trifluoromethyl iodobenzene (320.84 mg, 1.18 mmol, 169.76 uL, 2.00 eq), aqueous potassium hydroxide solution (4M, 737.19 uL, 5.00 eq) and bis(triphenylphosphine)palladium dichloride (20.70 mg, 29.49 umol, 0.05 eq) were added to a solution of 12 (278.00 mg, 589.75 umol, 1.00 eq) in 10 mL dimethyltetrahydrofuran. The reaction solution was stirred at 70 C. under nitrogen atmosphere for 12 hours, then 20 mL ethyl acetate was added. The mixture was filtered through celite, and the filtrate was washed twice with 30 mL saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by silica gel column chromatography (PE:EA=20:1 to 10:1) to give the product 16 (200.00 mg, crude product) as a yellow slime. MS (ESI, M+1): 490.2.
(25) Step B: Lithium hydroxide (97.85 mg, 4.09 mmol, 10.00 eq) was added to a solution of compound 16 (200.00 mg, 408.56 umol, 1.00 eq) in a mixed solvent of 2 mL methanol, 2 mL tetrahydrofuran and 2 mL water, the reaction solution was stirred at 30 C. for 1 hour. 10 mL water was added, then the mixture was adjusted to pH 5 with 1M hydrochloric acid and extracted twice with 10 mL ethyl acetate. The organic phase was combined, washed twice with 10 mL water, dried over anhydrous sodium sulfate, filtrated and concentrated to give a crude product, which was purified by preparative HPLC (formic acid system) to give the product I-4 (20.90 mg, 45.29 umol, yield 11.09%, purity 100%). MS (ESI, M+1): 462.2.
(26) .sup.1H NMR EW3644-177-P1B (400 MHz, DMSO-d.sub.6): 10.77 (s, 1H), 7.56 (d, J=8.0 Hz, 1H), 7.51-7.46 (m, 3H), 7.40 (d, J=8.8 Hz, 4H), 7.30 (d, J=8.0 Hz, 1H), 7.06 (t, J=8.0 Hz, 1H), 6.99 (t, J=7.2 Hz, 1H), 6.90 (d, J=8.4 Hz, 2H), 6.55 (d, J=1.6 Hz, 1H), 6.40 (d, J=16.0 Hz, 1 H), 2.75 (q, J=7.2 Hz 2 H), 0.98 (t, J=7.2 Hz, 3 H).
Embodiment 5
(27) ##STR00075##
(28) Step A: A solution of compound 12 (558.00 mg, 1.18 mmol, 1.00 eq), 2-trifluoromethyl iodobenzene (572.85 mg, 2.11 mmol, 1.80 eq), aqueous potassium hydroxide solution (4M, 1.64 mL, 5.60 eq) and dichlorobis(triphenylphosphine)palladium (24.64 mg, 35.10 umol, 0.03 eq) in 10 mL 2-methyltetrahydrofuran was purged with nitrogen for three times. The reaction solution was stirred at 80 C. under nitrogen atmosphere for 24 hours, then diluted with 30 mL water. The mixture was extracted twice with 50 mL ethyl acetate each time. The organic phase was combined, washed twice with 50 mL saturated brine each time, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to give the compound 17 as a yellow oil (200.00 mg), which was directly used in the next step.
(29) Step B: Lithium hydroxide monohydrate (85.71 mg, 2.04 mmol, 5.00 eq) was added to a solution of compound 17 (200.00 mg, 408.56 umol, 1.00 eq) in a mixed solvent of 6 mL tetrahydrofuran, 20 mL methanol and 6 mL water, the reaction solution was stirred at 30 C. for 3 hours. The reaction solution was concentrated to 1 mL, then adjusted to pH 5-6 with 3 mol/L hydrochloric acid, extracted twice with 50 mL dichloromethane each time. The organic phase was combined, dried over anhydrous sodium sulfate, filtrated and concentrated to give a crude product, which was purified by preparative chromatography (formic acid) to give the product I-5 (44.90 mg, 97.30 umol, yield 22.45%). MS [ESI, M+1]: 462.1.
(30) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.79 (s, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.60-7.50 (m, 2H), 7.46-7.29 (m, 6H), 7.12-6.90 (m, 4H), 6.59-6.50 (m, 1H), 6.39 (d, J=16.0 Hz, 1H), 2.90-2.79 (m, 1H), 2.48-2.41 (m, 1H), 0.95 (t, J=7.6 Hz, 3H).
Embodiment 6
(31) ##STR00076##
(32) Step A: A solution of compound 12 (279.00 mg, 590.95 umol, 1.00 eq), 2-iodothiophene (247.74 mg, 1.18 mmol, 120.26 uL, 2.00 eq), aqueous potassium hydroxide solution (4M, 737.19 uL, 5.00 eq), dichlorobis(triphenylphosphine)palladium (20.70 mg, 29.49 umol, 0.05 eq) in 10 mL 2-methyltetrahydrofuran was purged with nitrogen for three times and stirred at 70 C. for 12 hours. The reaction solution was diluted with 20 mL ethyl acetate, filtered, and the filtrate was washed twice with 30 mL saturated brine each time, dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography (PE:EA=20:1 to 10:1) to give the compound 18 (200.00 mg) as a yellow jelly, which was directly used in the next step.
(33) Step B: Lithium hydroxide (112.03 mg, 4.68 mmol, 10.00 eq) was added to a solution of compound 18 (200.00 mg, 467.77 umol, 1.00 eq) in a mixed solvent of 2 mL methanol, 2 mL tetrahydrofuran and 2 mL water, the reaction solution was stirred at 30 C. for 1 hour. Then 10 mL water was added, the mixture was adjusted to pH 5 with 1M hydrochloric acid, extracted twice with 10 mL ethyl acetate each time. The organic phase was combined, dried over anhydrous sodium sulfate, filtrated and concentrated to give a crude product, which was purified by preparative HPLC (formic acid system) to give the product I-6 (12.40 mg, 30.83 umol, yield 6.59%, purity 99.34%). MS [ESI, M+1]: 400.1.
(34) .sup.1H NMR (400 MHz, DMSO-d.sub.6): 10.72 (s, 1H), 7.52 (d, J=8.0 Hz, 3H), 7.48 (d, J=16.0 Hz, 1H), 7.40 (dd, J=5.2 Hz, J=1.2 Hz, 1H), 7.28 (d, J=8.4 Hz, 1H), 7.09 (d, J=8.4 Hz, 2H), 7.04 (t, J=6.0 Hz, 1H), 6.97 (t, J=6.8 Hz, 1H), 6.91 (dd, J=3.6 Hz, J=4.8 Hz, 1H), 6.87 (dd, J=0.8 Hz, J=3.6 Hz, 1H), 6.49 (d, J=1.6 Hz, 1H), 6.46 (d, J=16.0 Hz, 1 H), 2.71 (q, J=7.2 Hz 2 H), 1.10 (t, J=7.2 Hz, 3 H); MS(ESI, M+1): 400.1.
Embodiment 7
(35) ##STR00077##
(36) Intermediate 13 was prepared by the method of reaction procedure 1.
(37) Step A: Compound 13 (200.00 mg, 409.32 umol, 1.00 eq) and N-bromosuccinimide (87.42 mg, 491.18 umol, 1.20 eq) were dissolved in 5 mL dichloromethane. The reaction solution was stirred at 15 C. under nitrogen atmosphere for 1 hour, then concentrated and purified by silica gel chromatography to give the product 19 as a yellow solid (150.00 mg, 271.32 umol, yield 66.28%).
(38) Step C: Compound 19 (80.00 mg, 144.70 umol, 1.00 eq), lithium hydroxide (30.36 mg, 723.51 umol, 5.00 eq) were dissolved in a mixed solvent of 1 mL methanol, 1 mL tetrahydrofuran and 0.5 mL water. The reaction solution was stirred at 30 C. under nitrogen atmosphere for 3 hours. Then the reaction solution was adjusted to pH 5-6 with hydrochloric acid (1 mol/L) and filtered to give a crude product, which was purified by preparative chromatography (formic acid system) to give the product I-7 (61.00 mg, 116.23 umol, yield 80.32%, purity 99.29%). MS [ESI, M+1]=526.0.
(39) .sup.1H NMR (400 MHz, DMSO-d.sub.6) : 11.66 (s, 1H), 7.51-7.35 (m, 6H), 7.30-7.24 (m, 1H), 7.22-7.09 (m, 3H), 7.01 (d, J=8.4 Hz, 2H), 6.39 (d, J=16.0 Hz, 1H), 2.47-2.32 (m, 2H), 0.88 (t, J=7.6 Hz, 3H).
Embodiment 8
(40) ##STR00078##
(41) Step A: Compound I-1 (150.00 mg, 336.39 umol, 1.00 eq) and N-chlorosuccinimide (53.90 mg, 403.67 umol, 1.20 eq) were dissolved in 5 mL acetonitrile. The reaction solution was stirred at 15 C. under nitrogen atmosphere for 6 hours. After completion of the reaction, the reaction solution was concentrated and purified by preparative chromatography (formic acid system) to give the product I-8 (10.00 mg, 21.56 umol, yield 11.78%, purity 97.65%). MS [ESI, M+1].sup.+=480.1.
(42) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 11.51 (s, 1H), 7.50 (d, J=8.0 Hz, 1H), 7.47-7.37 (m, 5H), 7.29 (dd, J=6.4, 8.4 Hz, 1H), 7.22-7.09 (m, 3H), 7.01 (d, J=8.4 Hz, 2H), 6.41 (d, J=16.4 Hz, 1H), 2.49-2.36 (m, 2H), 0.89 (t, J=7.6 Hz, 3H).
Embodiment 9
(43) ##STR00079## ##STR00080##
(44) Step A: N,N-dimethylpyridine (406.83 mg, 3.33 mmol, 0.05 eq) and Boc.sub.2O (15.00 g, 68.73 mmol, 15.79 mL, 1.03 eq) were added to a solution of 20 (9.00 g, 66.60 mmol, 1.00 eq) in 180 mL dichloromethane. The reaction solution was stirred at 15 C. for 12 hours, then quenched with 100 mL water at 0 C., extracted three times with 210 mL ethyl acetate. The organic phase was combined, washed with 200 mL saturated brine, dried over anhydrous sodium sulfate, filtrated, concentrated and purified by column chromatography to give the product 21 as a colorless oil (15.40 g, 65.46 mmol, yield 98.29%).
(45) Step B: Lithium diisopropylamide (2M, 70.14 mL, 2.00 eq) was added dropwise to a solution of 21 (16.50 g, 70.14 mmol, 1.00 eq) in 300 mL tetrahydrofuran at 70 C. under nitrogen atmosphere. The reaction solution was stirred at 70 C. for 30 minutes, then cyanogen bromide (22.29 g, 210.42 mmol, 15.48 mL, 3.00 eq) was added to the reaction solution. The reaction solution was stirred at 15 C. for 12 hours, then quenched with 50 mL saturated aqueous ammonium chloride solution, diluted with 50 mL water and extracted with 300 mL ethyl acetate. The organic phase was combined, washed twice with 200 mL saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by column chromatography to give the crude product 22 as a yellow oil (14.10 g, 44.88 mmol, yield 63.99%).
(46) Step C: Compound 22 (12.00 g, 38.20 mmol, 1.00 eq), 1-trimethylsilylbutyne (9.65 g, 76.40 mmol, 2.00 eq), cuprous iodide (145.50 mg, 764.00 umol, 0.02 eq), cesium carbonate (16.30 g, 50.04 mmol, 1.31 eq), 1,1-bis(diphenylphosphino)ferrocene (423.55 mg, 764.00 umol, 0.02 eq) and palladium acetate (171.53 mg, 764.00 umol, 0.02 eq) were dissolved in 100 mL N,N-dimethylacetamide, the reaction system was purged with nitrogen for three times, and the reaction solution was stirred at 80 C. under nitrogen atmosphere for 6 hours. After completion of the reaction, the reaction solution was quenched with 100 mL water, diluted with 100 mL ethyl acetate, filtered through celite, the filtrate was extracted three times with 300 mL ethyl acetate. The organic phase was combined, washed with 300 mL brine for three times, dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to give the product 23 as a yellow oil (6.00 g, 20.88 mmol, yield 54.66%).
(47) Step D: Compound 23 (6.00 g, 20.88 mmol, 1.00 eq), diboron pinacol ester (5.30 g, 20.88 mmol, 1.00 eq), tetrakis(triphenylphosphine)platinum (1.30 g, 1.04 mmol, 0.05 eq) were dissolved in 100 mL 2-methyltetrahydrofuran, the reaction system was purged with nitrogen for three times. The reaction solution was stirred at 80 C. under nitrogen atmosphere for 4 hours. After completion of the reaction, the reaction solution was quenched with 50 mL water at 0 C., then diluted with 100 mL ethyl acetate, filtered through celite, and the filtrate was extracted three times with 300 mL ethyl acetate. The organic phase was combined, washed with 100 mL brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to give the product 24 as a yellow oil (4.00 g, 7.17 mmol, yield 34.33%, purity 97%). MS [ESI, M+1].sup.+=542.3.
(48) Step E: Compound 24 (515.46 mg, 923.75 umol, 1.00 eq), 2-chloro-4-fluoroiodobenzene (236.89 mg, 923.75 umol, 1.00 eq), cesium carbonate (601.95 mg, 1.85 mmol, 2.00 eq), dichlorobis(triphenylphosphine)palladium (64.84 mg, 92.38 umol, 0.10 eq) and 1.2 mL water were dissolved in 30 mL 2-methyltetrahydrofuran, and the reaction system was purged with nitrogen for three times. The reaction solution was stirred at 75 C. under nitrogen atmosphere for 12 hours. After completion of the reaction, the reaction solution was quenched with 30 mL water at 0 C., then diluted with 100 mL ethyl acetate, filtered through celite, and the filtrate was extracted three times with 90 mL ethyl acetate. The organic phase was combined, washed with 50 mL brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography to give the product 25 as a yellow oil (500.00 mg), which was used directly in the next step. MS [ESI, M+1].sup.+=446.0.
(49) Step F: Compound 25 (200.00 mg, 450.73 umol, 1.00 eq), compound 5 (292.84 mg, 901.46 umol, 2.00 eq), dichlorobis(triphenylphosphine)palladium (63.27 mg, 90.15 umol, 0.20 eq) and aqueous potassium hydroxide (4M, 631.02 uL, 5.60 eq) were dissolved in 5 mL 2-methyltetrahydrofuran, and the reaction system was purged with nitrogen for three times. The reaction solution was stirred at 80 C. under nitrogen atmosphere for 12 hours. After completion of the reaction, the reaction solution was quenched with 30 mL water at 0 C., then diluted with 30 mL ethyl acetate, filtered through celite, and the filtrate was extracted three times with 90 mL ethyl acetate. The organic phase was combined, washed with 50 mL brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography to give the crude product 26 (90.00 mg), which was used directly in the next step. MS [ESI, M+1].sup.+=492.2.
(50) Step G: Compound 26 (90.00 mg, 182.94 umol, 1.00 eq) and lithium hydroxide (38.38 mg, 914.70 umol, 5.00 eq) were dissolved in a mixed solvent of 2 mL methanol, 1 mL tetrahydrofuran and 0.5 mL water. The reaction solution was stirred at 25 C. for 12 hours, then concentrated to 3 mL, diluted with 20 mL water. The mixture was adjusted to pH 5-6 with hydrochloric acid (1 mol/L) and extracted with 200 mL dichloromethane. The organic phase was combined, concentrated and purified by preparative chromatography (formic acid system) to give the product I-9 (10.00 mg, 21.56 umol, yield 11.78%). MS [ESI, M+1].sup.+=464.2.
(51) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.82 (s, 1H), 7.56 (dd, J=5.6, 8.8 Hz, 1H), 7.48-7.32 (m, 4H), 7.25 (dd, J=6.4, 8.4 Hz, 1H), 7.12 (dt, J=2.4, 8.4 Hz, 1H), 7.05 (dd, J=2.0, 10.0 Hz, 1H), 6.96 (d, J=8.0 Hz, 2H), 6.91-6.81 (m, 1H), 6.60 (d, J=1.6 Hz, 1H), 6.41 (d, J=14.4 Hz, 1H), 2.70-2.59 (m, 2H), 1.00 (t, J=7.2 Hz, 3H).
Embodiment 10
(52) ##STR00081##
(53) Intermediate 30 was prepared using a similar procedure to that described in Embodiment 9, but using indole as the raw material.
(54) Step A: Cesium carbonate (6.52 g, 20.00 mmol, 4.00 eq) and 1,2-dibromoethane (4.70 g, 25.00 mmol, 1.89 mL, 5.00 eq) were added to a solution of p-iodophenol (1.10 g, 5.00 mmol, 1.00 eq) in acetonitrile (20.00 mL), the reaction solution was stirred at 85 C. for 24 hours, then concentrated under reduced pressure to remove the solvent, diluted with 20 mL water and extracted three times with 30 mL ethyl acetate each time. The organic phase was combined, washed with 30 mL saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by silica gel column chromatography (PE/EA=100/1 to 20/1) to give the product 27 (1.40 g, 4.28 mmol, yield 85.64%).
(55) Step B: Cesium carbonate (3.28 g, 10.08 mmol, 3.00 eq) and compound 28 (506.30 mg, 4.03 mmol, 1.20 eq) were added to a solution of compound 27 (1.10 g, 3.36 mmol, 1.00 eq) in acetonitrile (20.00 mL), the reaction solution was stirred at 85 C. for 12 hours, then diluted with 50 mL water and extracted three times with 50 mL ethyl acetate each time. The organic phase was combined, washed twice with 50 mL saturated brine each time, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (DCM/MeOH=100/1 to 10/1) to give the product 29 (700.00 mg, 2.09 mmol, yield 62.16%).
(56) Step C: Aqueous potassium hydroxide solution (4M, 190.17 uL, 4.00 eq) was added to a mixture of compound 30 (201.61 mg, 190.17 umol, 1.00 eq), compound 29 (127.47 mg, 380.34 umol, 2.00 eq) and dichlorobis(triphenylphosphine)palladium (133.48 mg, 190.17 umol, 1.00 eq), the reaction solution was purged with nitrogen and then stirred at 80 C. under nitrogen atmosphere for 12 hours. The reaction solution was concentrated to give a crude product, which was purified by preparative high performance liquid chromatography (formic acid system) to give the product II-1 (2.80 mg, 5.49 umol, yield 2.89%, purity 99.5%). MS(ESI, M+1): 507.1. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.65 (s, 1H), 8.32 (s, 1H), 7.55 (d, J=7.6 Hz, 1H), 7.36 (dd, J=2.4, 8.8 Hz, 1H), 7.30 (d, J=8.0 Hz, 1H), 7.24-7.18 (m, 1H), 7.11 (dt, J=2.4, 8.4 Hz, 1H), 7.08-7.02 (m, 1H), 7.02-6.95 (m, 1H), 6.84 (d, J=8.8 Hz, 2H), 6.64 (d, J=8.8 Hz, 2H), 6.53 (s, 1H), 4.54 (d, J=6.4 Hz, 1H), 4.42 (d, J=6.4 Hz, 1H), 3.79 (t, J=5.2 Hz, 2H), 3.27 (t, J=7.2 Hz, 2H), 2.96 (t, J=6.4 Hz, 2H), 2.71-2.60 (m, 5H), 0.99 (t, J=7.2 Hz, 3H).
Embodiment 11
(57) ##STR00082##
(58) Step A: Compound 25 (340.00 mg, 625.18 umol, 1.00 eq) and compound 29 (419.07 mg, 1.25 mmol, 2.00 eq), dichlorobis(triphenylphosphine)palladium (219.41 mg, 312.59 umol, 0.50 eq) and aqueous potassium hydroxide solution (4M, 875.26 uL, 5.60 eq) were dissolved in 10 mL 2-methyltetrahydrofuran, the reaction system was purged with nitrogen for three times. The reaction solution was stirred at 80 C. under nitrogen atmosphere for 12 hours. After completion of the reaction, the reaction solution was cooled to 0 C., quenched with 30 mL water, diluted with 30 mL ethyl acetate, filtered through celite, and the filtrate was extracted three times with 20 mL ethyl acetate. The organic phase was combined, washed with 20 mL brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by preparative thin layer chromatography to give the crude product 31 as a yellow oil (110.00 mg, crude), which was used directly in the next step. MS [ESI, M56]=625.2.
(59) Step B: Trifluoroacetic acid (770.00 mg, 6.75 mmol, 500.00 uL, 42.22 eq) was added to a solution of compound 31 (90.00 mg, 152.37 umol, 1.00 eq) in 33 mL dichloromethane, and the reaction solution was stirred at 16 C. for 3 hours. The reaction solution was adjusted to pH 7 to 8 with saturated aqueous sodium bicarbonate solution, diluted with 20 mL water, extracted twice with 100 mL dichloromethane each time. The organic phase was combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by preparative chromatography (formic acid system) to give the product II-2 (10.00 mg, 18.86 umol, yield 12.38%, purity 99.0%).
(60) MS [ESI, M+1].sup.+=525.2.
(61) .sup.1H NMR (400 MHz, DMSO-d.sub.6) =10.71 (s, 1H), 8.29 (br. s., 1H), 7.55 (dd, J=5.6, 8.4 Hz, 1H), 7.35 (dd, J=2.4, 8.8 Hz, 1H), 7.21 (dd, J=6.4, 8.4 Hz, 1H), 7.13-7.02 (m, 2H), 6.91-6.77 (m, 3H), 6.65 (d, J=8.8 Hz, 2H), 6.56 (d, J=1.2 Hz, 1H), 4.54 (d, J=6.4 Hz, 1H), 4.42 (d, J=6.4 Hz, 1H), 3.79 (t, J=5.2 Hz, 2H), 3.28 (t, J=6.8 Hz, 2H), 2.97 (t, J=6.4 Hz, 2H), 2.71-2.62 (m, 5H), 0.99 (t, J=7.6 Hz, 3H).
Embodiment 12
(62) ##STR00083## ##STR00084##
(63) Step A: N,N-dimethylpyridine (135.61 mg, 1.11 mmol, 0.05 eq) and Boc.sub.2O (4.89 g, 22.42 mmol, 1.01 eq) were added to a solution of 32 (3.00 g, 22.20 mmol, 1.00 eq) in 30 mL dichloromethane. The reaction solution was stirred at 10-15 C. for 12 hours, then quenched with 50 mL aqueous ammonium chloride solution at 0 C., extracted twice with 25 mL dichloromethane each time. The organic phase was combined, washed with 50 mL saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give the product 33 as a white oil (5.20 g, 22.10 mmol, yield 99.57%).
(64) Step B: Lithium diisopropylamide (2M, 14.96 mL, 1.10 eq) was added dropwise to a solution of 33 (6.40 g, 27.21 mmol, 1.00 eq) in 60 mL tetrahydrofuran at 70 C., after stirring at 70 C. for 30 minutes, a solution of cyanogen bromide (8.64 g, 81.62 mmol, 6.00 mL, 3.00 eq) in 20 mL tetrahydrofuran was added dropwise to the above solution. The reaction solution was stirred at 15 C. for 12 hours. Then 40 mL water and 40 mL ethyl acetate were successively added, and the mixture was partitioned in a separating funnel. The organic phase was washed three times with 120 mL water (40 mL each time), dried over anhydrous sodium sulfate, concentrated and filtrated to give a crude product, which was purified by column chromatography to give the product 34 as a yellow oil (7.80 g, 24.83 mmol, yield 91.25%).
(65) Step C: Compound 34 (7.80 g, 24.83 mmol, 1.00 eq), 1-trimethylsilylbutyne (4.70 g, 37.24 mmol, 1.50 eq), cuprous iodide (236.43 mg, 1.24 mmol, 0.05 eq), cesium carbonate (10.52 g, 32.28 mmol, 1.30 eq), 1,1-bis(diphenylphosphino)ferrocene (688.23 mg, 1.24 mmol, 0.05 eq) and palladium acetate (278.72 mg, 1.24 mmol, 0.05 eq) were dissolved in 80 mL N,N-dimethylacetamide, the reaction system was purged with nitrogen for three times, and the reaction solution was stirred at 80-90 C. under nitrogen atmosphere for 48 hours. After completion of the reaction, the reaction solution was diluted with 100 mL water, extracted twice with 100 mL ethyl acetate each time. The organic phase was combined, washed twice with 100 mL brine each time, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to give the product 35 as a yellow oil (4.00 g, 13.92 mmol, yield 56.07%).
(66) Step D: Compound 35 (4.00 g, 13.92 mmol, 1.00 eq), diboron pinacol ester (3.57 g, 14.06 mmol, 1.01 eq), tetrakis(triphenylphosphine)platinum (865.42 mg, 696.06 umol, 0.05 eq) were dissolved in 50 mL 2-methyltetrahydrofuran, the reaction system was purged with nitrogen for three times. The reaction solution was stirred at 70-80 C. under nitrogen atmosphere for 5 hours, then concentrated under reduced pressure to give a crude product, which was purified by column chromatography to give the product 36 as a yellow solid (2.10 g, 3.88 mmol, yield 27.87%).
(67) Step E: Compound 36 (2.10 g, 3.88 mmol, 1.00 eq), 2-chloro-4-fluoroiodobenzene (994.93 mg, 3.88 mmol, 1.00 eq), cesium carbonate (2.53 g, 7.76 mmol, 2.00 eq) and dichlorobis(triphenylphosphine)palladium (272.32 mg, 387.98 umol, 0.10 eq) were dissolved in 10 mL 2-methyltetrahydrofuran, and the reaction system was purged with nitrogen for three times. The reaction solution was stirred at 70-80 C. under nitrogen atmosphere for 12 hours, then concentrated under reduced pressure, diluted with 50 mL water, extracted twice with 50 mL ethyl acetate each time. The organic phase was combined, washed with 30 mL saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give the product 37 as a yellow oil (400.00 mg).
(68) Step F: Compound 37 (200.00 mg, 367.76 umol, 1.00 eq), compound 29 (246.52 mg, 735.52 umol, 2.00 eq), dichlorobis(triphenylphosphine)palladium (25.81 mg, 36.78 umol, 0.10 eq) and aqueous potassium hydroxide solution (4M, 514.86 uL, 5.60 eq) were dissolved in 5 mL 2-methyltetrahydrofuran, and the reaction system was purged with nitrogen for three times. The reaction solution was stirred at 80 C. under nitrogen atmosphere for 12 hours, then concentrated under reduced pressure, and purified by preparative thin layer chromatography to give the crude product 38 (100.00 mg), which was used directly in the next step.
(69) Step G: Trifluoroacetic acid (770.00 mg, 6.75 mmol, 500.00 uL, 42.22 eq) was added to a solution of compound 38 (100.00 mg, 159.97 umol, 1.00 eq) in 10 mL dichloromethane, and the reaction solution was stirred at 10-16 C. for 12 hours. The reaction solution was adjusted to pH 7-8 with aqueous sodium bicarbonate solution, diluted with 20 mL water, extracted twice with 25 mL dichloromethane each time. The organic phase was combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by HPLC (formic acid system) to give the product II-3 (20.00 mg, 38.05 umol, yield 23.79%, purity 99.89%). MS (ESI, M+1): 525.2.
(70) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 5=10.75 (s, 1H), 7.34 (dd, J=2.8, 9.2 Hz, 1H), 7.31-7.25 (m, 2H), 7.20 (dd, J=6.4, 8.8 Hz, 1H), 7.13-7.07 (m, 1H), 6.88 (dt, J=2.6, 9.2 Hz, 1H), 6.83-6.80 (m, 2H), 6.65-6.62 (m, 2H), 6.52 (d, J=1.6 Hz, 1H), 4.53 (d, J=6.2 Hz, 1H), 4.41 (d, J=6.2 Hz, 1H), 3.78 (t, J=5.6 Hz, 2H), 3.28-3.23 (m, 3H), 2.94 (t, J=6.4 Hz, 2H), 2.68-2.59 (m, 4H), 0.97 (t, J=7.2 Hz, 3H).
Embodiment 13
(71) ##STR00085## ##STR00086##
(72) Step A: Boron tribromide (6.74 g, 26.91 mmol, 2.59 mL, 3.00 eq) was added dropwise to a solution of compound 39 (2.00 g, 8.97 mmol, 1.00 eq) in dichloromethane (20.00 mL). After completion of the addition, the reaction solution was stirred at 25 C. for 12 hours, then quenched by the dropwise addition of 40 mL methanol, concentrated to give a crude product, which was purified by silica gel column chromatography to give the product 40 as a brown oil (2 g).
(73) Step B: Cesium carbonate (2.34 g, 7.18 mmol, 1.00 eq) and 1,2-dibromoethane (6.74 g, 35.90 mmol, 2.71 mL, 5.00 eq) were added to a solution of compound 40 (1.50 g, 7.18 mmol, 1.00 eq) in acetonitrile (10.00 mL), the reaction solution was stirred at 80 C. for 12 hours, then poured into 50 mL water, extracted twice with 50 mL ethyl acetate each time. The organic phase was combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (PE/EA=10/1) to give the product 41 as a yellow oil (1.40 g, 4.43 mmol, yield 61.72%).
(74) Step C: Cesium carbonate (2.06 g, 6.32 mmol, 4.00 eq) and compound 28 (768.02 mg, 4.74 mmol, 3.00 eq, 2HCl) were added to a solution of compound 41 (500.00 mg, 1.58 mmol, 1.00 eq) in acetonitrile (20.00 mL), the reaction solution was stirred at 70 C. for 1 hour, then diluted with 30 mL water, extracted with 30 mL ethyl acetate, dried over anhydrous sodium sulfate, filtered, concentrated to give a crude product, which was purified by preparative TLC (PE/EA=1/2) to give the product 42 as a yellow solid (400.00 mg, 1.23 mmol, yield 78.10%).
(75) Step D: Potassium phosphate (2M, 1.64 mL, 4.00 eq) and Ad2nBuP Biphenyl (54.62 mg, 81.77 umol, 0.10 eq) were added to a solution of compound 42 (430.00 mg, 817.72 umol, 1.00 eq) and compound 30 (318.07 mg, 981.26 umol, 1.20 eq) in 8 mL tetrahydrofuran. The reaction solution was stirred at 80 C. under nitrogen atmosphere, then diluted with 50 mL water, extracted twice with 50 mL ethyl acetate each time for 36 hours, dried over anhydrous sodium sulfate, filtered, concentrated to give a crude product, which was purified by silica gel column chromatography (PE/EA=15/1 to 0/1) and preparative high performance liquid chromatography (formic acid system) to give the product II-4 (530.00 ug, 0.98 umol, yield 0.12%). MS(ESI, M+1): 543.1.
(76) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.70 (s, 1H), 7.52 (d, J=8.0 Hz, 1H), 7.35-7.31 (m, 2H), 7.26-7.20 (m, 1H), 7.15-7.10 (m, 1H), 7.10-7.03 (m, 1H), 7.01-6.95 (m, 1H), 6.50 (t, J=8.0 Hz, 3H), 4.54 (d, J=6.4 Hz, 1H), 4.42 (d, J=6.4 Hz, 1H), 3.84 (t, J=5.2 Hz, 2H), 3.28 (t, J=7.2 Hz, 2H), 2.95 (t, J=5.2 Hz, 2H), 2.90-2.75 (m, 2H), 2.74-2.60 (m, 3H), 1.03 (t, J=7.2 Hz, 3H).
Embodiment 14
(77) ##STR00087##
(78) Step A: N-chlorosuccinimide (390.71 mg, 2.93 mmol, 1.10 eq) was added to a solution of the crude product 17 (1.30 g) in 10 mL acetonitrile, the reaction solution was stirred at 15 C. under nitrogen atmosphere for 5 hours, then quenched with 40 mL water, extracted three times with 120 mL ethyl acetate (40 mL each time). The organic phase was combined, washed with 100 mL saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product 44 (1.4 g, crude).
(79) Step B: Lithium hydroxide (319.96 mg, 13.35 mmol, 5.00 eq) was added to a solution of the crude product 44 (1.40 g) in a mixed solvent of 10 mL tetrahydrofuran, 10 mL methanol and 2.5 mL water, the reaction solution was stirred at 30 C. for 1 hour, then concentrated to 1 mL under reduced pressure, adjusted to pH 5-6 with 3M hydrochloric acid and filtered. The filter cake was dried and purified by preparative chromatography (formic acid system) to give the product I-10 (691.00 mg, 1.38 mmol, yield 51.66%, purity 99%).
(80) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 12.32 (br s, 1H), 11.48 (s, 1H), 7.71 (d, J=7.6 Hz, 1H), 7.64-7.57 (m, 1H), 7.53-7.45 (m, 2H), 7.43-7.30 (m, 5H), 7.22-7.10 (m, 2H), 6.97 (d, J=8.4 Hz, 2H), 6.37 (d, J=16.0 Hz, 1H), 2.57-2.51 (m, 1H), 2.38-2.29 (m, 1H), 0.86 (t, J=7.6 Hz, 3H). MS [ESI, M+1]: 496.1.
Embodiment 15
(81) ##STR00088##
(82) Step A: N-bromosuccinimide (520.77 mg, 2.93 mmol, 1.10 eq) was added to a solution of the crude product 17 (1.30 g, 2.66 mmol, 1.00 eq) in 10 mL acetonitrile, the reaction solution was stirred at 15 C. under nitrogen atmosphere for 5 hours, then quenched with 40 mL water, extracted three times with 120 mL ethyl acetate (40 mL each time). The organic phase was combined, washed with 100 mL saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product 45 (1.6 g, crude), which was directly used in the next step.
(83) Step B: Lithium hydroxide (336.50 mg, 14.05 mmol, 5.00 eq) was added to a solution of the crude product 45 (1.60 g, 2.8 mmol, 1.00 eq) in a mixed solvent of 10 mL tetrahydrofuran, 10 mL methanol and 2.5 mL water, the reaction solution was stirred at 30 C. for 1 hour. The reaction solution turned from deep yellow to brown, then concentrated to 1 mL under reduced pressure, adjusted to pH 5-6 with 3M hydrochloric acid and filtered. The filter cake was purified by preparative chromatography (formic acid system) and then lyophilized to give the product I-11 as a pale yellow solid (811.00 mg, 1.49 mmol, yield 52.88%, purity 99%).
Embodiment 16
(84) ##STR00089##
(85) Step A: Diboron pinacol ester (329.61 mg, 1.30 mmol, 1.10 eq) and tetrakis(triphenylphosphine)platinum (73.53 mg, 59.09 umol, 0.05 eq) were added to a solution of compound 10 (200.00 mg, 1.18 mmol, 1.00 eq) in 2-methyltetrahydrofuran (5.00 mL), the reaction solution was purged three times with nitrogen and stirred at 70 C. under nitrogen atmosphere for 5 hours. After the reaction solution was cooled to 0 C., compound 5 (285.58 mg, 945.27 umol, 0.80 eq), cesium carbonate (769.97 mg, 2.36 mmol, 2.00 eq), 0.4 mL water, 5 mL 2-methyltetrahydrofuran and dichlorobis(triphenylphosphine)palladium (41.47 mg, 59.08 umol, 0.05 eq) were added, then the reaction solution was purged three times with nitrogen and stirred at 20 C. under nitrogen atmosphere for 12 hours. Then 4-fluoro-2-trifluoromethyl iodobenzene (678.60 mg, 2.34 mmol, 2.00 eq), aqueous potassium hydroxide solution (4M, 1.46 mL, 5.00 eq), dichlorobis(triphenylphosphine)palladium (41.06 mg, 58.50 umol, 0.05 eq) and 10 mL 2-methyltetrahydrofuran were added to the above solution, which was subsequently purged three times with nitrogen. After stirring at 70 C. under nitrogen atmosphere for 12 hours, the reaction solution was filtered, and the filtrate was directly concentrated and then purified by silica gel column chromatography (PE:EA=40:1 to 15:1) to give the compound 57 as a yellow solid (350.00 mg, 628.94 umol, yield 53.76%, purity 91.2%).
(86) Step B: Lithium hydroxide monohydrate (263.90 mg, 6.29 mmol, 10.00 eq) was added to a solution of compound 57 (350.00 mg, 628.94 umol, 1.00 eq) in a mixed solvent of 3 mL tetrahydrofuran, 3 mL methanol and 3 mL water, the reaction solution was stirred at 35 C. for 1 hour. 10 mL water was added to the reaction solution, then the mixture was adjusted to pH 5 with 1 mol/L hydrochloric acid and extracted twice with 10 mL ethyl acetate each time. The organic phase was combined, washed twice with 10 mL water each time, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative high performance liquid chromatography (formic acid system) to give the product I-12 (108.80 mg, 226.01 umol, yield 35.94%, purity 99.6%).
(87) .sup.1H NMR (400 MHz, DMSO-d.sub.6): 10.78 (brs, 1H), 9.57-9.54 (m, 2H), 7.48-7.38 (m, 5H), 7.31 (d, J=7.6 Hz, 1H), 7.06 (dt, J=6.8 Hz, J=0.8 Hz, 1H), 6.99 (dt, J=6.8 Hz, J=0.8 Hz, 1H), 6.93 (d, J=8.0 Hz, 2H), 6.53 (d, J=1.6 Hz, 1H), 6.40 (d, J=16.0 Hz, 1H), 2.85-2.78 (m, 1 H), 2.47-2.39 (m, 1 H), 0.95 (t, J=7.6 Hz, 3 H). MS(ESI, M+1): 480.2.
Embodiment 17
(88) ##STR00090## ##STR00091##
(89) Step A: Diboron pinacol ester (329.61 mg, 1.30 mmol, 1.10 eq) and tetrakis(triphenylphosphine)platinum (73.53 mg, 59.09 umol, 0.05 eq) were added to a solution of compound 10 (200.00 mg, 1.18 mmol, 1.00 eq) in 2-methyltetrahydrofuran (5.00 mL), the reaction solution was purged three times with nitrogen and stirred at 70 C. under nitrogen atmosphere for 5 hours. After the reaction solution was cooled to 0 C., compound 5 (285.58 mg, 945.27 umol, 0.80 eq), cesium carbonate (769.97 mg, 2.36 mmol, 2.00 eq), 0.4 mL water, 5 mL 2-methyltetrahydrofuran and dichlorobis(triphenylphosphine)palladium (41.47 mg, 59.08 umol, 0.05 eq) were added, then the reaction solution was purged three times with nitrogen and stirred at 20 C. under nitrogen atmosphere for 12 hours. Then 2-chloro-4-trifluoromethyl iodobenzene (717.09 mg, 2.34 mmol, 2.00 eq), aqueous potassium hydroxide (4M, 1.46 mL, 5.00 eq), dichlorobis(triphenylphosphine)palladium (41.06 mg, 58.50 umol, 0.05 eq) and 10 mL 2-methyltetrahydrofuran were added to the reaction solution, which was subsequently purged three times with nitrogen. After stirring at 70 C. under nitrogen atmosphere for 12 hours, the reaction solution was filtered through celite, concentrated and purified by silica gel column chromatography (PE:EA=40:1 to 15:1) to give the compound 47 as a yellow jelly (738.00 mg), which was directly used in the next step.
(90) Step B: Lithium hydroxide monohydrate (591.64 mg, 14.10 mmol, 10.00 eq) was added to a solution of compound 47 (738.80 mg, 1.41 mmol, 1.00 eq) in a mixed solvent of 3 mL methanol, 3 mL tetrahydrofuran and 3 mL water, the reaction solution was stirred at 35 C. for 1 hour. 10 mL water was added to the reaction solution, then the resulting solution was adjusted to pH 5 with 1 mol/L hydrochloric acid, extracted twice with 10 mL ethyl acetate each time. The organic phase was combined, washed twice with 10 mL water each time, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative high performance liquid chromatography (formic acid system) to give the product I-13 (135.30 mg, 267.64 umol, yield 18.98%, purity 98.10%).
(91) .sup.1H NMR (400 MHz, DMSO-d.sub.6): 10.77 (brs, 1H), 7.81 (brs, 1H), 7.62 (dd, J=8.0 Hz, J=1.2 Hz, 1H), 7.57 (d, J=8.0 Hz, 1H), 7.47-7.40 (m, 4 H), 7.31 (d, J=8.0 Hz, 1H), 7.07 (dt, J=8.0 Hz, J=1.2 Hz, 1H), 7.02-6.96 (m, 3H), 6.61 (d, J=1.6 Hz, 1H), 6.41 (d, J=16.0 Hz, 1H), 2.75-2.62 (m, 2 H), 1.00 (t, J=7.6 Hz, 3H); MS(ESI, M+1): 496.2.
Embodiment 18
(92) ##STR00092## ##STR00093##
(93) Step A: Diboron pinacol ester (1.65 g, 6.50 mmol, 1.10 eq) and tetrakis(triphenylphosphine)platinum (367.64 mg, 295.50 umol, 0.05 eq) were added to a solution of compound 10 (1.09 g, 5.91 mmol, 1.00 eq) in 2-methyltetrahydrofuran (20.00 mL), the reaction solution was purged three times with nitrogen and stirred at 70 C. under nitrogen atmosphere for 5 hours. After the reaction solution was cooled to 0 C., compound 5 (1.43 g, 4.73 mmol, 0.80 eq), cesium carbonate (3.85 g, 11.82 mmol, 2.00 eq), 1.2 mL water, 10 mL 2-methyltetrahydrofuran and dichlorobis(triphenylphosphine)palladium (207.34 mg, 295.50 umol, 0.05 eq) were added, then the reaction solution was purged three times with nitrogen and stirred at 20 C. under nitrogen atmosphere for 12 hours. Then 4-chloro-2-trifluoromethyl iodobenzene (3.63 g, 11.84 mmol, 2.00 eq), aqueous potassium hydroxide solution (4M, 7.40 mL, 5.00 eq) and dichlorobis(triphenylphosphine)palladium (207.72 mg, 296.00 umol, 0.05 eq) were added to the reaction solution, which was subsequently purged three times with nitrogen. After stirring at 70 C. under nitrogen atmosphere for 12 hours, the reaction solution was filtered, concentrated and purified by silica gel column chromatography (PE:EA=50:1 to 5:1) to give the compound 48 as a yellow jelly (2.60 g), which was directly used in the next step.
(94) Step B: Lithium hydroxide monohydrate (2.08 g, 49.60 mmol, 10.00 eq) was added to a solution of compound 48 (2.60 g, 4.96 mmol, 1.00 eq) in a mixed solvent of 20 mL methanol, 20 mL tetrahydrofuran and 20 mL water, the reaction solution was stirred at 35 C. for 1 hour. 60 mL water was added to the reaction solution, then the mixture was adjusted to pH 5 with 1 mol/L hydrochloric acid, extracted twice with 60 mL ethyl acetate each time. The organic phase was combined, washed twice with 50 mL water each time, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative high performance liquid chromatography (formic acid system) to give the product I-14 (735.10 mg, 1.48 mmol, yield 29.77%, purity 99.6%).
(95) .sup.1H NMR (400 MHz, DMSO-d.sub.6): 12.33 (brs, 1H), 10.77 (brs, 1H), 7.73 (d, J=2.0 Hz, 1H), 7.65 (dd, J=8.4 Hz, J=2.0 Hz, 1H), 7.55 (d, J=8.0 Hz, 1H), 7.45-7.37 (m, 4H), 7.31 (d, J=8.0 Hz, 1H), 7.06 (t, J=7.2 Hz, 1H), 6.99 (t, J=7.2 Hz, 1H), 6.94 (d, J=8.0 Hz, 2H), 6.53 (d, J=1.2 Hz, 1H), 6.40 (d, J=16.0 Hz, 1H), 2.87-2.78 (m, 1 H), 2.47-2.41 (m, 1 H), 0.94 (t, J=7.6 Hz, 3 H). MS (ESI, M+1 ): 496.0.
Embodiment 19
(96) ##STR00094## ##STR00095##
(97) Step A: 3-Bromo-2-cyanothiophene (195.55 mg, 1.04 mmol, 2.00 eq) and aqueous potassium hydroxide solution (4M, 520.00 uL, 4.00 eq) were added to a solution of compound 12 (300.00 mg, 519.95 umol, 1.00 eq) in 2-methyltetrahydrofuran (10.00 mL), after the reaction system was purged with nitrogen for three times, dichlorobis(triphenylphosphine)palladium (18.25 mg, 26.00 umol, 0.05 eq) was added. The reaction solution was stirred at 70 C. under nitrogen atmosphere for 12 hours, then diluted with 20 mL ethyl acetate, filtered through celite. The filtrate was washed three times with 20 mL brine each time, dried over anhydrous sodium sulfate, filtered and concentrated to give the compound 49 as a brown jelly (400.00 mg, crude product).
(98) Step B: Lithium hydroxide monohydrate (370.86 mg, 8.84 mmol, 10.00 eq) was added to a solution of compound 49 (400.00 mg, 883.84 umol, 1.00 eq) in a mixed solvent of 2 mL methanol, 2 mL tetrahydrofuran and 2 mL water, the reaction solution was stirred at 45 C. for 1 hour, then adjusted to pH 5 with 1 mol/L hydrochloric acid. 20 mL ethyl acetate was added, then washed twice with 20 mL brine each time, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative high performance liquid chromatography (formic acid system) to give the intermediate 50 (75.00 mg, 163.0716.12 umol, yield 18.45%, purity 92.3%).
(99) Step C: N-chlorosuccinimide (26.13 mg, 195.68 umol, 1.20 eq) was added to a solution of compound 50 (75.00 mg, 163.07 umol, 1.00 eq) in acetonitrile (5.00 mL), the reaction solution was stirred at 30 C. for 14 hours, then concentrated and purified by preparative high performance liquid chromatography (formic acid system) to give I-15 (7.40 mg, 16.12 umol, yield 9.89%, purity 100%).
(100) .sup.1H NMR (400 MHz, DMSO-d.sub.6): 11.46 (brs, 1H), 8.03 (d, J=5.2 Hz, 1H), 7.51 (d, J=7.6 Hz, 1H), 7.44 (d, J=8.4 Hz, 2H), 7.40 (d, J=16.4 Hz, 1H), 7.37 (d, J=8.0 Hz, 1H), 7.31 (d, J=5.2 Hz, 1H), 7.19 (dt, J=7.2 Hz, J=1.2 Hz, 1H), 7.13 (dt, J=8.0 Hz, J=1.2 Hz, 1H), 6.92 (d, J=8.4 Hz, 2H), 6.42 (d, J=16.0 Hz, 1H), 2.50-2.46 (m, 2H), 0.94 (t, J=7.6 Hz, 3 H). MS (ESI, M+1, M+23): 459.0, 481.1.
Embodiment 20
(101) ##STR00096##
(102) Step A: Bis(diphenylphosphino)ferrocene dichloropalladium (7.94 mg, 10.86 umol, 0.05 eq) was added to a solution of 51 (100.00 mg, 217.14 umol, 1.00 eq), 2,5-dichloro-3-bromothiophene (70.50 mg, 304.00 umol, 1.40 eq), aqueous sodium hydroxide solution (4M, 217.14 uL, 4.00 eq) in 2-methyltetrahydrofuran (10 mL) in a 100 mL three-necked flask. After the reaction system was purged with nitrogen under reduced pressure, the reaction solution was stirred at 70 C. (internal temperature 60 C.) for 12 hours, then turned from yellow to brown. 30 mL ethyl acetate was added to the reaction solution, and the resulting mixture was filtered through celite. The filtrate was washed three times with 30 mL saturated brine each time, dried over anhydrous sodium sulfate, filtered and concentrated to give the product 53 as a brown solid (200.00 mg, crude product), which was directly used in the next step.
(103) Step B: Lithium hydroxide monohydrate (169.04 mg, 4.03 mmol, 10.00 eq) was added to a solution of compound 53 (200.00 mg, 402.86 umol, 1.00 eq) in a mixed solvent of 2 mL methanol, 2 mL tetrahydrofuran and 2 mL water in a 100 mL single-neck flask, the reaction solution was stirred at 45 C. for 1 hour and turned from yellow to brown, then adjusted to pH 1 with 1 mol/L hydrochloric acid. 30 mL ethyl acetate and 30 mL water were added, then the resulting mixture was partitioned. The organic phase was washed twice with 30 mL saturated brine each time, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative high performance liquid chromatography (formic acid system) to give I-17 (36.00 mg, 76.71 umol, yield 19.04%, purity 99.8%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): 10.73 (brs, 1H), 7.70 (d, J=3.6 Hz, 1H), 7.53 (d, J=8.0 Hz, 3H), 7.45 (d, J=16.0 Hz, 1H), 7.29 (d, J=8.0 Hz, 1H), 7.10-7.04 (m, 3H), 6.98 (t, J=7.6 Hz, 1H), 6.91 (d, J=3.6 Hz, 1H), 6.54 (brs, 1H), 6.47 (d, J=16.0 Hz, 1H), 2.75 (q, J=7.2 Hz, 2H), 2.43 (s, 3H), 1.11 (t, J=7.6 Hz, 3 H). MS (ESI, M+1): 468.0.
Embodiment 21
(104) ##STR00097##
(105) Step A: Bis(diphenylphosphino)ferrocene dichloropalladium (7.94 mg, 10.86 umol, 0.05 eq) was added to a solution of 51 (100.00 mg, 217.14 umol, 1.00 eq), 3-bromo-2-cyanothiophene (57.17 mg, 304.00 umol, 1.40 eq), aqueous sodium hydroxide solution (4M, 217.14 uL, 4.00 eq) in 2-methyltetrahydrofuran (10 mL) in a 50 mL three-necked flask. After the reaction system was purged with nitrogen under reduced pressure, the reaction solution was stirred at 70 C. for 12 hours, then turned from yellow to brown. 30 mL ethyl acetate was added to the reaction solution, and the mixture was filtered through celite. The filtrate was washed three times with 30 mL saturated brine each time, dried over anhydrous sodium sulfate, filtered and concentrated to give the product 54 as a brown solid (200.00 mg, crude product), which was directly used in the next step.
(106) Step B: Lithium hydroxide monohydrate (185.43 mg, 4.42 mmol, 10.00 eq) was added to a solution of compound 54 (200.00 mg, 402.86 umol, 1.00 eq) in a mixed solvent of 2 mL methanol, 2 mL tetrahydrofuran and 2 mL water in a 100 mL single-neck flask, the reaction solution was stirred at 45 C. for 1 hour and turned from yellow to brown, then adjusted to pH 1 with 1 mol/L hydrochloric acid. 30 mL ethyl acetate and 30 mL water were added, then the mixture was partitioned. The organic phase was washed twice with 30 mL saturated brine each time, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative high performance liquid chromatography (formic acid system) to give I-18 (24.70 mg, 57.95 umol, yield 13.11%, purity 99.6%).
(107) .sup.1H NMR (400 MHz, DMSO-d.sub.6): 10.78 (brs, 1H), 7.99 (d, J=5.2 Hz, 1H), 7.57 (d, J=8.0 Hz, 1H), 7.49-7.45 (m, 3H), 7.31-7.28 (m, 2H), 7.07 (t, J=7.2 Hz, 1H), 7.00 (t, J=7.2 Hz, 1H), 6.93 (d, J=8.4 Hz, 2H), 6.58 (d, J=1.2 Hz, 1H), 6.45 (d, J=16.0 Hz, 1H), 2.71 (q, J=7.6 Hz, 2H), 1.02 (t, J=7.6 Hz, 3 H). MS (ESI, M+1): 425.1.
Embodiment 22
(108) ##STR00098##
(109) Step A: 19 (50.00 mg, 90.44 umol, 1.00 eq), 2,4,6-trimethylboroxine (34.06 mg, 271.32 umol, 37.84 uL, 3.00 eq), cesium carbonate (88.40 mg, 271.32 umol, 3.00 eq) and 3 mL dioxane were added to a single-neck round bottom flask. After the reaction system was purged with nitrogen, Pd(dppf)Cl.sub.2.DCM (7.39 mg, 9.04 umol, 0.1 eq) was added, and the reaction system was purged with nitrogen again. The yellow emulsible mixture was stirred at 120 C. for 1 hour and turned from bright yellow to dark yellow. The reaction solution was filtered and concentrated to give a crude product, which was purified by preparative TLC (PE:EA=5:1) to give the intermediate 55 (25.00 mg, 51.23 umol, yield 56.65%). MS (ESI, M+1): 488.1.
(110) Step B: Lithium hydroxide monohydrate (6.45 mg, 153.69 umol, 3.00 eq) was added to a solution of compound 55 (25.00 mg, 51.23 umol, 1.00 eq) in a mixed solvent of 2 mL tetrahydrofuran, 2 mL methanol and 2 mL water, the colorless transparent solution was stirred at 20 C. for 72 hours, then concentrated under reduced pressure, diluted with 10 mL water, adjusted to pH 1-2 with 1 mol/L hydrochloric acid, extracted three times with 20 mL ethyl acetate, washed with 20 mL saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative TLC (PE:EA=3:1) to give the product I-19 (20.00 mg, 43.44 umol, yield 84.79%, purity 99.9%).
(111) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.80 (s, 1H), 8.30 (s, 1H), 7.49 (d, J=7.6 Hz, 1H), 7.43-7.32 (m, 5H), 7.30 (d, J=8.0 Hz, 1H), 7.18 (dt, J=2.4, 8.5 Hz, 1H), 7.07 (dt, J=1.2, 7.6 Hz, 1H), 7.03-6.95 (m, 3H), 6.39 (d, J=16.0 Hz, 1H), 2.43 (q, J=7.6 Hz, 2H), 2.29 (s, 3H), 0.86 (t, J=7.6 Hz, 3H); MS (ESI, M+1): 460.1.
Embodiment 23
(112) ##STR00099## ##STR00100##
(113) Step A: 51 (101.68 mg, 221.57 umol, 1.00 eq), 5-chloro-2-iodobenzonitrile (70.05 mg, 265.88 umol, 1.20 eq), Pd(dppf)Cl.sub.2 (8.11 mg, 11.08 umol, 0.05 eq) and 2-methyltetrahydrofuran (5.00 mL) were added to a three-necked round bottom flask, after the reaction system was purged with nitrogen, 4M sodium hydroxide (221.57 uL, 4.00 eq) was added, and the reaction system was purged with nitrogen again. The reaction solution was heated to 68 C. and stirred for 10 hours, then filtered. The filter cake was washed with 10 mL ethyl acetate and the filtrate was concentrated to give intermediate 63 (125.00 mg), which was directly used in the next step. MS (ESI, M+1): 481.2.
(114) Step B: 63 (125.00 mg, 259.88 umol, 1.00 eq), tetrahydrofuran (2.00 mL) and methanol (2.00 mL) were added to a single-neck round bottom flask, followed by addition of lithium hydroxide monohydrate (32.71 mg, 779.64 umol, 3.00 eq) and water (2.00 mL). The reaction solution was stirred at 30 C. for 12 hours, then concentrated to remove the solvent. 5 mL water was added to the reaction solution, and the resulting mixture was adjusted to pH 3 with 2M hydrochloric acid, extracted twice with 20 mL ethyl acetate each time. The organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative high performance liquid chromatography (formic acid system) to give the product I-21 (32.00 mg, 70.51 umol, yield 27.13%, purity 99.8%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) =10.77 (s, 1H), 7.88 (d, J=2.0 Hz, 1H), 7.75 (dd, J=2.0, 8.4 Hz, 1H), 7.61 (dd, J=8.0, 12.0 Hz, 2H), 7.49-7.37 (m, 3H), 7.31 (br d, J=8.0 Hz, 1H), 7.08 (br t, J=7.2 Hz, 1H), 7.04-6.99 (m, 1H), 6.94 (br d, J=8.0 Hz, 2H), 6.62 (s, 1H), 6.44 (br d, J=16.0 Hz, 1H), 2.95-2.57 (m, 2H), 1.03 (br t, J=7.2 Hz, 3H); MS (ESI, M+1): 453.1.
(115) Step C:
(116) I-21 (80.00 mg, 176.63 umol, 1.00 eq), 3 mL dichloromethane, 6 mL acetonitrile and NCS (25.94 mg, 194.29 umol, 1.10 eq) were added to a single-neck round bottom flask under nitrogen atmosphere. The reaction solution was stirred at 25 C. for 1 hour, then quenched with 10% sodium thiosulphate (20 mL). 20 mL dichloromethane was added, and the mixture was partitioned, the aqueous phase was extracted three times with 10 mL dichloromethane each time. The organic phase was combined, washed with 20 mL saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative high performance liquid chromatography (formic acid system) to give the product I-22 (25.00 mg, 50.99 umol, yield 28.87%, purity 99.4%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) =11.47 (br s, 1H), 7.94 (br s, 1H), 7.78 (br d, J=8.0 Hz, 1H), 7.60-7.49 (m, 2H), 7.48-7.36 (m, 4H), 7.25-7.09 (m, 2H), 6.96 (br d, J=7.2 Hz, 2H), 6.41 (br d, J=15.6 Hz, 1H), 2.61-2.53 (m, 2H), 0.93 (br t, J=6.8 Hz, 3H); MS (ESI, M+1): 487.0.
Embodiment 25
(117) ##STR00101##
(118) Step A: Compound 19 (50.00 mg, 85.92 umol, 1.00 eq) and CuCN (23.08 mg, 257.75 umol, 3.00 eq) were dissolved in NMP (2.00 mL). The reaction solution was purged with nitrogen, stirred at 180 C. for 2 hours under microwave irradiation and turned from green to black. After completion of the reaction, water (20 mL) was added, and the mixture was adjusted to pH 10 with aqueous NaHCO.sub.3 (2 mL) and extracted with ethyl acetate (20 mL*3). The organic phase was combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative TLC (PE:EA=5:1) to give the product 64 as a yellow oil (10.00 mg, 16.43 umol, yield 19.13%, purity 82%). MS (ESI, M+1): 499.1.
(119) Step B: Compound 64 (80.00 mg, 160.33 umol, 1.00 eq) was dissolved in a mixed solvent of tetrahydrofuran (1.50 mL), ethanol (1.50 mL) and water (1.50 mL), followed by addition of LiOH.H.sub.2O (20.18 mg, 480.99 umol, 3.00 eq), then the reaction solution was stirred at 30 C. for 12 hours. The reaction was incomplete, then sodium hydroxide (9.62 mg, 240.50 umol, 1.50 eq) and ethanol (1.50 mL) were added to the reaction solution. The yellow reaction solution was stirred at 45 C. for 3 hours. After completion of the reaction, the reaction solution was concentrated to give a crude product, then water (1 mL) and 3 mol/L HCl were added to the crude product to adjust pH to 4. Then a solid precipitated, which was isolated by filtration. The solid was purified by preparative chromatography (formic acid system) to give the product I-23 (25.90 mg, 54.45 umol, yield 33.96%, purity 99%). MS (ESI, M+1): 471.0.
(120) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 12.36 (br s, 1H), 7.65 (d, J=6.8 Hz, 1H), 7.52 (d, J=8.0 Hz, 1H), 7.48-7.41 (m, 3H), 7.37 (br d, J=16.0 Hz, 1H), 7.34-7.25 (m, 3H), 7.25-7.17 (m, 1H), 7.01 (d, J=8.4 Hz, 2H), 6.41 (br d, J=16.4 Hz, 1H), 2.46-2.40 (m, 1H), 2.46-2.40 (m, 1H), 0.94 (t, J=7.6 Hz, 3H).
Embodiment 26
(121) ##STR00102## ##STR00103##
(122) Step A: Compound 12 (340.91 mg, 636.42 umol, 1.00 eq) was dissolved in 2-methyltetrahydrofuran (10.00 mL), then potassium hydroxide (4M, 890.99 uL, 5.60 eq) and 1-chloro-4-iodobenzene (273.16 mg, 1.15 mmol, 1.80 eq) was added. The reaction solution was purged with nitrogen for three times and stirred at 80 C. for 12 hours. After completion of the reaction, water (30 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL*3). The organic phase was combined, washed with saturated brine (30 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by column chromatography (PE:EA=1:0-20:1) to give the intermediate 65 (280.00 mg, 614.08 umol, yield 96.49%). MS (ESI, M+1): 456.0.
(123) Step B: Compound 65 (280.00 mg, 564.95 umol, 1.00 eq) was dissolved in acetonitrile (8.00 mL), NCS (82.98 mg, 621.44 umol, 1.10 eq) was added in one portion. The reaction solution was stirred at 20 C. for 2 hours. After completion of the reaction, the reaction solution was concentrated to give a crude product. Then water (5 mL) was added to the crude product, and the mixture was extracted with ethyl acetate (5 mL*4). The organic phase was combined, washed with saturated brine (10 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product 66 (230.00 mg, 389.26 umol, yield 68.90%, purity 83%), which was directly used in the next step without further purification. MS (ESI, M+1): 490.0.
(124) Step C: Compound 66 (230.00 mg, 389.26 umol, 1.00 eq) was dissolved in a mixed solvent of water (3.00 mL), methanol (6.00 mL) and tetrahydrofuran (6.00 mL), then LiOH.H.sub.2O (81.67 mg, 1.95 mmol, 5.00 eq) was added. The reaction solution was stirred at 20 C. for 12 hours. After completion of the reaction, the reaction solution was concentrated, adjusted to pH 4 with 3 mol/L HCl (1 mL), filtered to give a solid, which was dissolved in 1 mL DMF and purified by preparative chromatography (formic acid system) to give the product I-24 as a yellow solid (41.00 mg, 81.58 umol, yield 20.96%, purity 92%), but the purity was not enough. The product was purified by alkaline preparative chromatography, but the purity was still not enough, then purified by preparative TLC (dichloromethane:ethyl acetate=10:1) to give the product I-24 (9.70 mg, 20.56 umol, yield 25.20%, purity 98%).
(125) 1H NMR (400 MHz, DMSO-d.sub.6) 11.47 (s, 1H), 7.50 (d, J=7.6 Hz, 1H), 7.47-7.30 (m, 6H), 7.25-7.16 (m, 3H), 7.16-7.10 (m, 1H), 6.93 (d, J=8.4 Hz, 2H), 6.40 (d, J=16.0 Hz, 1H), 2.48-2.40 (m, 2H), 0.90 (t, J=7.6 Hz, 3H). MS (ESI, M+1): 462.0.
Embodiment 27
(126) ##STR00104## ##STR00105##
(127) Step A: Trifluoromethanesulfonic anhydride (19.07 g, 67.59 mmol, 11.15 mL, 3.00 eq) was slowly added dropwise to a solution of oxindole (3.00 g, 22.53 mmol, 1.00 eq) and triethylamine (22.80 g, 225.30 mmol, 31.23 mL, 10.00 eq) in 100 mL 2-methyltetrahydrofuran at 78 C. under nitrogen atmosphere in a reaction flask. The reaction solution was stirred for more than 2 hours, and turned from colorless to orange. Then 4-methyl-1-pentyne (3.70 g, 45.06 mmol, 5.29 mL, 2.00 eq), triethylamine (22.80 g, 225.30 mmol, 31.23 mL, 10.00 eq), tetrakis(triphenylphosphine)platinum (2.60 g, 2.25 mmol, 0.10 eq) and cuprous iodide (429.08 mg, 2.25 mmol, 0.10 eq) were added. The reaction solution was purged with nitrogen, warmed to 15 C. and stirred under nitrogen atmosphere for 12 hours. The gray suspension turned into a black solution. The reaction solution was quenched by saturated aqueous ammonium chloride solution (300 mL), extracted three times with 100 mL ethyl acetate each time. The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (EA:PE=0-5%) to give the compound 68 (4.90 g, 14.88 mmol, yield 66.04%).
(128) Step B: Potassium carbonate (3.78 g, 27.32 mmol, 2.00 eq) was added to a solution of compound 68 (4.50 g, 13.66 mmol, 1.00 eq) in 50 mL tetrahydrofuran and 50 mL methanol. The reaction solution was stirred at 25 C. for 12 hours and the colorless mixture turned from yellow to brown. 200 mL ethyl acetate and 200 mL water were added to the reaction solution. The mixture was partitioned in a separatory funnel. The organic phase was washed three times with 100 mL water each time, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the compound 69 (2.60 g, 13.18 mmol, yield 96.48%).
(129) Step C: Tetrakis(triphenylphosphine)platinum (126.29 mg, 101.50 umol, 0.05 eq) was added to a solution of compound 69 (400.00 mg, 2.03 mmol, 1.00 eq) and diboron pinacol ester (515.50 mg, 2.03 mmol, 1.00 eq) in 10 mL 2-methyltetrahydrofuran. The reaction solution was stirred at 70 C. for 5 hours and gradually turned from yellow to brown. The reaction solution of compound 70 in 10 mL 2-methyltetrahydrofuran was directly used in the next step.
(130) Step D: Compound 5 (490.63 mg, 1.62 mmol, 0.80 eq), 10 mL 2-methyltetrahydrofuran and 3 mL water were added to the solution of compound 70 (915.96 mg, 2.03 mmol, 1.00 eq) in 10 mL 2-methyltetrahydrofuran. Then cesium carbonate (1.32 g, 4.06 mmol, 2.00 eq) and bis(triphenylphosphine)palladium dichloride (142.49 mg, 203.00 umol, 0.10 eq) were quickly added to the mixture at 0 C. The reaction solution was stirred at 25 C. under nitrogen atmosphere for 12 hours and gradually turned from yellow to brown. The reaction solution of compound 71 in 20 mL 2-methyltetrahydrofuran was directly used in the next step.
(131) Step E: 2-Chloro-4-fluoro-iodobenzene (621.61 mg, 2.42 mmol, 1.20 eq) and 10 mL 2-methyltetrahydrofuran were added to the solution of compound 71 (1.01 g, 2.02 mmol, 1.00 eq) in 20 mL 2-methyltetrahydrofuran, then aqueous potassium hydroxide solution (4M, 2.53 mL, 5.00 eq) and bis(triphenylphosphine)palladium dichloride (70.89 mg, 101.00 umol, 0.05 eq) were added at 25 C. The reaction solution was stirred at 70 C. for 12 hours, then filtered, and 30 mL ethyl acetate and 30 mL water were added. The mixture was partitioned with a separatory funnel. The organic phase was washed with 30 mL brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (PE:EA=1:0 to 20:1) to give the intermediate 72 (500.00 mg, crude product).
(132) Step F: Lithium hydroxide monohydrate (417.91 mg, 9.96 mmol, 10.00 eq) was added to a solution of compound 72 (500.00 mg, 995.98 umol, 1.00 eq) in tetrahydrofuran (5.00 mL), methanol (5.00 mL) and water (5.00 mL) at 25 C., The mixture was stirred at 40 C. for 1 hour and turned from yellow to brown, then adjusted to pH 3 with 1M HCl, and 20 mL water was added. The mixture was extracted twice with 20 mL ethyl acetate each time. The organic phase was combined, washed twice with 20 mL brine each time, dried over anhydrous sodium sulfate, filtered, concentrated to give a crude product, which was purified by preparative high performance liquid chromatography (formic acid system) to give the product I-27 (150.00 mg, 307.62 umol, yield 30.89%, purity 97.2%).
(133) .sup.1H NMR (400 MHz, DMSO-d.sub.6): 10.78 (brs, 1H), 7.56 (d, J=7.6 Hz, 1H), 7.48-7.38 (m, 3H), 7.34 (dd, J=2.4 Hz, J=9.2 Hz, 1H), 7.31-7.25 (m, 2H), 7.11 (dt, J=2.4 Hz, J=8.4 Hz, 1H), 7.08-7.03 (m, 1H), 7.01-6.96 (m, 3H), 6.52 (d, J=1.2 Hz, 1H), 6.40 (d, J=16.0 Hz, 1H), 2.62-2.53 (m, 2H), 1.55-1.45 (m, 1H), 0.87 (d, J=6.8 Hz, 3H), 0.84 (d, J=6.8 Hz, 3H). MS(ESI, M+1): 474.1.
Embodiment 28
(134) ##STR00106## ##STR00107##
(135) Step A: NaOH (4M, 443.13 uL, 4.00 eq) was added to a solution of compound 51 (200.00 mg, 443.13 umol, 1.00 eq), 4-chloro-1-iodo-2-toluene (156.63 mg, 620.38 umol, 1.40 eq), Pd(dppf)Cl.sub.2 (16.21 mg, 22.16 umol, 0.05 eq) in 12 mL 2-methyltetrahydrofuran, the reaction solution was purged with nitrogen for three times, then stirred at 65 C. for 12 hours, gradually turned from yellow to black. 20 mL water was added to the above solution, and the mixture was extracted three times with ethyl acetate (20 mL*3). The organic phase was combined, washed with 20 mL saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (400 mesh, PE:EA=1:0 to 20:1) to give the compound 73 (170.00 mg, 325.53 umol, yield 73.46%, purity 90%).
(136) Step B: NCS (38.35 mg, 287.23 umol, 1.00 eq) was added to a solution of compound 73 (150.00 mg, 287.23 umol, 1.00 eq) in acetonitrile (6.00 mL) and dichloromethane (14.00 mL). The yellow solution was stirred at 30 C. for 1.5 hours. Then 30 mL water was added to the reaction solution, and the mixture was extracted twice with ethyl acetate (30 mL*2). The organic phase was combined, washed twice with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (PE:EA=1:0 to 10:1) to give the intermediate 74 (170.00 mg, crude).
(137) Step C: LiOH.H.sub.2O (70.70 mg, 1.69 mmol, 5.00 eq) was added to a solution of compound 74 (170.00 mg, 337.00 umol, 1.00 eq) in water (2.00 mL), tetrahydrofuran (2.00 mL) and ethanol (2.00 mL). The yellow solution was stirred at 30 C. for 1.5 hours, then concentrated under reduced pressure to give a residue. 1 mL water was added to the residue, the resulting solution was adjusted to pH 4 with 3M hydrochloric acid. A solid precipitated, which was isolated by filtration and then dissolved in 3 mL DMF and purified by preparative high performance liquid chromatography (formic acid system) to give the product I-28 (46.50 mg, 96.63 umol, yield 28.67%, purity 99%).
(138) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 11.46 (s, 1H), 7.50 (d, J=7.6 Hz, 1H), 7.46-7.37 (m, 4H), 7.31-7.23 (m, 2H), 7.22-7.09 (m, 2H), 7.00-6.91 (m, 3H), 6.40 (d, J=16.0 Hz, 1H), 2.45 (q, J=7.6 Hz, 2H), 2.27 (s, 3H), 0.90 (t, J=7.6 Hz, 3H). MS (ESI, M+1): 476.0.
Embodiment 29
(139) ##STR00108##
(140) Sodium hydroxide (34.87 mg, 871.64 umol, 4.00 eq) was added to a mixture of compound 51 (100.00 mg, 217.91 umol, 1.00 eq), 2-bromo-3-chlorotoluene (62.69 mg, 305.07 umol, 1.40 eq) and 2-methyltetrahydrofuran (10.00 mL) in a single-neck flask, followed by addition of 1,1-bis(diphenylphosphino)ferrocene palladium chloride (7.97 mg, 10.90 umol, 0.05 eq) under nitrogen atmosphere. The reaction solution was stirred at 70 C. for 16 hours. After completion of the reaction, the reaction solution was adjusted to pH 1 with hydrochloric acid (3M) at room temperature, then extracted with ethyl acetate (20 mL). The organic phase was concentrated to give a crude product, which was purified by preparative liquid phase chromatography (prep. HPLC: formic acid) to give the product I-29 (5.30 mg, 11.79 umol, yield 5.41%, purity 98.3%).
(141) MS (ESI, M+1): 442.0.
(142) .sup.1H NMR (400 MHz, DMSO-d.sub.6) =10.80 (s, 1H), 8.40 (s, 1H), 7.57 (d, J=7.3 Hz, 1H), 7.39-7.27 (m, 5H), 7.16-7.11 (m, 1H), 7.09-7.04 (m, 2H), 7.03-6.97 (m, 3H), 6.61 (d, J=1.4 Hz, 1H), 6.38 (d, J=15.9 Hz, 1H), 2.62-2.56 (m, 2H), 2.16 (s, 3H), 1.00 (t, J=7.5 Hz, 3H).
Embodiment 30
(143) ##STR00109## ##STR00110##
(144) Step A: Compound 12 (100.00 mg, 212.13 umol, 1.00 eq) and 6-bromo-1,4-benzodioxan (68.42 mg, 318.19 umol, 42.76 uL, 1.50 eq) were weighed and dissolved in 2-methyltetrahydrofuran (4.00 mL) under nitrogen atmosphere, followed by addition of potassium hydroxide (66.65 mg, 1.19 mmol, 5.60 eq) (dissolved in 500.00 uL water) and dichlorobis(triphenylphosphine)palladium(II) (7.44 mg, 10.61 umol, 0.05 eq), the reaction solution was stirred at 75 C. for 16 hours. After completion of the reaction, the reaction solution was filtered through silica gel (100-200 mesh), the silica gel was washed with ethyl acetate (30 mL), the filtrate was combined and concentrated to give the crude product 75 as a dark brown oil (210.00 mg), which was directly used in the next step.
(145) Step B: Lithium hydroxide monohydrate (36.75 mg, 875.78 umol, 2.00 eq) was added to a solution of the crude product 75 (210.00 mg) in tetrahydrofuran (4.00 mL), methanol (1.00 mL) and water (1.00 mL), the reaction solution was stirred at 25 C. for 4 hours. After completion of the reaction, the reaction solution was adjusted to pH 1 with hydrochloric acid (3M) at room temperature, then extracted with ethyl acetate (30 mL). The organic phase was washed with saturated brine (10 mL*2), concentrated to give a crude product, which was purified by preparative liquid phase chromatography (prep. HPLC: formic acid) to give the product I-30 (27.95 mg, 61.67 umol, yield 14.08%, purity 99.63%).
(146) MS (ESI, M+1): 452.0.
(147) .sup.1H NMR (400 MHz, DMSO-d.sub.6) =10.72 (s, 1H), 7.53 (d, J=7.8 Hz, 1H), 7.50-7.40 (m, 3H), 7.28 (d, J=8.0 Hz, 1H), 7.09-6.90 (m, 4H), 6.72-6.64 (m, 2H), 6.55 (dd, J=1.9, 8.3 Hz, 1H), 6.47 (d, J=1.4 Hz, 1H), 6.42 (d, J=16.1 Hz, 1H), 4.19 (s, 4H), 2.60 (q, J=7.2 Hz, 2H), 0.96 (t, J=7.3 Hz, 3H).
Embodiment 31
(148) ##STR00111##
(149) Compound I-30 (160.00 mg, 287.85 umol, 1.00 eq) was weighed and dissolved in acetonitrile (2.00 mL), N-chlorosuccinimide (38.44 mg, 287.85 umol, 1.00 eq) was added in portions, the reaction solution was stirred at 25 C. for 2 hours. After completion of the reaction, a saturated aqueous sodium thiosulfate solution (3 mL) was added to the reaction solution, the mixture was stirred for a few minutes, then extracted with dichloromethane (30 mL*1). The organic phase was concentrated to give a crude product, which was purified by preparative liquid phase chromatography (prep. HPLC: formic acid) to give the product I-31 as a yellow solid (25.82 mg, 51.84 umol, yield 18.01%, purity 97.56%).
(150) .sup.1H NMR (400 MHz, DMSO-d.sub.6) =11.43 (s, 1H), 8.40 (s, 1H), 7.48 (d, J=7.9 Hz, 1H), 7.41-7.33 (m, 4H), 7.20-7.10 (m, 2H), 6.94 (d, J=8.3 Hz, 2H), 6.77-6.68 (m, 2H), 6.66-6.61 (m, 1H), 6.38 (d, J=15.9 Hz, 1H), 4.21 (s, 4H), 2.39 (q, J=7.3 Hz, 2H), 0.89 (t, J=7.4 Hz, 3H); MS (ESI, M+1): 486.1.
Embodiment 32
(151) ##STR00112## ##STR00113##
(152) Step A: Compound 12 (100.00 mg, 212.13 umol, 1.00 eq) and 4-iodoanisole (74.47 mg, 318.19 umol, 1.50 eq) were weighed and dissolved in 2-methyltetrahydrofuran (4.00 mL) under nitrogen atmosphere, followed by addition of potassium hydroxide (66.65 mg, 1.19 mmol, 5.60 eq) (dissolved in 500.00 uL water) and dichlorobis(triphenylphosphine)palladium(II) (7.44 mg, 10.61 umol, 0.05 eq), the reaction solution was stirred at 75 C. for 16 hours. After completion of the reaction, the reaction solution was filtered through silica gel (100-200 mesh), the silica gel was washed with ethyl acetate (30 mL*1), the filtrate was combined and concentrated to give a crude product as a dark brown oil, which was purified by silica gel plate (PE/EA=10/1) to give the intermediate 76 (50.00 mg).
(153) Step B: Lithium hydroxide monohydrate (25.09 mg, 597.90 umol, 15.00 eq) was added to a solution of the crude product 76 (50.00 mg) in a mixed solvent of tetrahydrofuran (2.00 mL), methanol (0.50 mL) and water (0.50 mL), the reaction solution was stirred at 25 C. for 4 hours. After completion of the reaction, the reaction solution was adjusted to pH 1 with hydrochloric acid (3M) at room temperature, then extracted with ethyl acetate (30 mL*1). The organic phase was concentrated to give the crude product I-32 as a yellow solid (68.00 mg).
(154) Step C: Compound I-32 (68.00 mg) was weighed and dissolved in acetonitrile (2.00 mL) and dichloromethane (2.00 mL), N-chlorosuccinimide (10.72 mg, 80.28 umol, 0.50 eq) was added in portions, the reaction solution was stirred at 25 C. for 2 hours. After completion of the reaction, a saturated aqueous sodium thiosulfate solution (3 mL) was added to the reaction solution, the resulting mixture was stirred for a few minutes, then extracted with dichloromethane (30 mL*1). The organic phase was concentrated to give a crude product, which was purified by preparative liquid phase chromatography (prep. HPLC: formic acid) to give the product I-33 (8.71 mg, 18.39 umol, yield 11.45%, purity 96.67%).
(155) MS (ESI, M+1): 458.1.
(156) .sup.1H NMR (400 MHz, DMSO-d.sub.6) =11.44 (s, 1H), 8.42 (s, 1H), 7.48 (d, J=7.7 Hz, 1H), 7.40-7.29 (m, 4H), 7.20-7.16 (m, 1H), 7.15-7.09 (m, 3H), 6.92 (d, J=8.2 Hz, 2H), 6.84 (d, J=8.7 Hz, 2H), 6.36 (d, J=15.9 Hz, 1H), 3.73 (s, 3H), 2.43 (m, J=7.5 Hz, 2H), 0.89 (t, J=7.3 Hz, 3H).
Embodiment 34
(157) ##STR00114## ##STR00115##
(158) Step A: Compound 12 (200.00 mg, 339.42 umol, 1.00 eq) and 4-bromo-3-chlorobenzonitrile (110.21 mg, 509.13 umol, 1.50 eq) were weighed and dissolved in 2-methyltetrahydrofuran (8.00 mL) under nitrogen atmosphere, followed by addition of potassium hydroxide (106.65 mg, 1.90 mmol, 5.60 eq) (dissolved in 1.00 mL water) and dichlorobis(triphenylphosphine)palladium(II) (11.91 mg, 16.97 umol, 0.05 eq), the reaction solution was stirred at 75 C. for 8 hours. After completion of the reaction, the reaction solution was concentrated to remove 2-methyltetrahydrofuran, extracted with ethyl acetate (30 mL*1), dried, concentrated to give a crude product, which was purified by preparative TLC (PE/EA=5/1), further by preparative liquid phase chromatography (prep. HPLC: formic acid) to to give the intermediate 77 (8.00 mg, 15.76 umol, yield 4.64%, purity 94.75%).
(159) Step B: Lithium hydroxide monohydrate (5.00 mg, 119.16 umol, 7.56 eq) was added to a solution of the compound 77 (8.00 mg) in a mixed solvent of tetrahydrofuran (2.00 mL), methanol (0.50 mL) and water (0.50 mL), the reaction solution was stirred at 25 C. for 4 hours. Under the same condition, additional lithium hydroxide monohydrate (5.00 mg, 119.16 umol, 7.56 eq) was added, and the reaction was allowed to continue for another 2 hours. After completion of the reaction, the reaction solution was adjusted to pH 1 with hydrochloric acid (4M) under an ice bath, then concentrated, extracted with ethyl acetate (15 mL*1). The organic phase was concentrated to give a crude product, which was purified by preparative liquid phase chromatography (prep. HPLC: formic acid) to give the product I-34 (3.60 mg, 7.88 umol, yield 50.02%, purity 99.18%).
(160) MS (ESI, M+1): 453.0.
(161) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.77 (s, 1H), 8.38 (s, 1H), 7.99 (d, J=1.5 Hz, 1H), 7.73 (dd, J=1.6, 7.9 Hz, 1H), 7.57 (d, J=7.8 Hz, 1H), 7.46 (d, J=8.1 Hz, 1H), 7.43-7.29 (m, 4H), 7.11-7.05 (m, 1H), 7.04-6.94 (m, 3H), 6.61 (d, J=1.5 Hz, 1H), 6.40 (d, J=16.0 Hz, 1H), 2.74-2.67 (m, 2H), 1.00 (t, J=7.5 Hz, 3H).
Embodiment 35
(162) ##STR00116##
(163) Compound I-17 (70.00 mg, 124.04 umol, 1.00 eq) was weighed and dissolved in acetonitrile (4.00 mL) and dichloromethane (4.00 mL), N-chlorosuccinimide (11.59 mg, 86.83 umol, 0.70 eq) was added in portions, the reaction solution was stirred at 25 C. for 2 hours. After completion of the reaction, a saturated aqueous sodium thiosulfate solution (2 mL) was added to the reaction solution, the mixture was stirred for a few minutes, then extracted with dichloromethane (30 mL*1). The organic phase was concentrated to give a crude product, which was purified by preparative TLC plate (dichloromethane/ethyl acetate=1/1, with a small amount of acetic acid) to give a yellow solid 2 (20.00 mg, 35.25 umol, yield 28.42%, purity 88.63%). The solid 2 was purified by preparative TLC (100% ethyl acetate), further by preparative liquid phase chromatography (formic acid) to give the product I-35 as a yellow solid (8.00 mg, 15.64 umol, yield 47.38%, purity 98.3%).
(164) .sup.1H NMR (400 MHz, CHLOROFORM-d) =7.88 (s, 1H), 7.72-7.64 (m, 2H), 7.36-7.29 (m, 3H), 7.28-7.21 (m, 2H), 7.05 (d, J=8.4 Hz, 2H), 6.72 (s, 1H), 6.38 (d, J=16.0 Hz, 1H), 2.53 (q, J=7.6 Hz, 2H), 1.05 (t, J=7.6 Hz, 3H); MS (ESI, M+1): 502.0.
Embodiment 36
(165) ##STR00117##
(166) Step A: Compound I-17 (93.00 mg, 198.55 umol, 1.00 eq) was weighed and dissolved in acetonitrile (4.00 mL) and dichloromethane (4.00 mL), N-chlorosuccinimide (24.74 mg, 138.98 umol, 0.70 eq) was added in portions, the reaction solution was stirred at 25 C. for 1 hour. After completion of the reaction, a saturated aqueous sodium thiosulfate solution (3 mL) was added to the reaction solution, the mixture was stirred for a few minutes, then extracted with dichloromethane (30 mL*1). The organic phase was concentrated to give a crude product, which was purified by preparative TLC (dichloromethane/ethyl acetate=1/1, with a small amount of acetic acid) to give the intermediate 78 (95.00 mg, 151.02 umol, yield 76.06%, purity 87%).
(167) Step B: Compound 78 (95.00 mg, 168.38 umol, 1.00 eq) was dissolved in N-methylpyrrolidone (2.00 mL) in a microwave tube, followed by addition of copper cyanide (31.06 mg, 346.86 umol, 75.76 uL, 2.06 eq), the reaction solution was stirred at 180 C. for 1.5 hours under microwave irradiation. Under the same condition, additional copper cyanide (30.16 mg, 336.76 umol, 73.56 uL, 2.00 eq) was added to the reaction solution and the reaction was allowed to continue for 2 hours. Under the same condition, additional copper cyanide (45.24 mg, 505.14 umol, 110.34 uL, 3.00 eq) was added to the reaction solution and the reaction was allowed to continue for 2 hours. After completion of the reaction, the reaction solution was poured into ice water (20 mL), extracted with ethyl acetate (50 mL*2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative liquid chromatography (prep. HPLC: formic acid) to give the product I-36 as a yellow solid (7.18 mg, 13.75 umol, yield 8.16%, purity 94.46%).
(168) .sup.1H NMR (400 MHz, DMSO-d.sub.6) =12.28 (s, 1H), 7.66 (d, J=7.1 Hz, 1H), 7.55 (d, J=8.3 Hz, 2H), 7.52-7.46 (m, 2H), 7.33-7.24 (m, 3H), 7.00 (d, J=8.3 Hz, 2H), 6.47 (d, J=16.0 Hz, 1H), 2.44-2.39 (m, 2H), 0.98 (t, J=7.4 Hz, 3H); MS (ESI, M+1): 493.0.
Embodiment 37
(169) ##STR00118## ##STR00119##
(170) Step A: Compound 12 (300.00 mg, 560.05 umol, 1.00 eq) and 2,4-dichloro-1-iodobenzene (275.11 mg, 1.01 mmol, 1.80 eq), 2-methyltetrahydrofuran (10.00 mL) and aqueous potassium hydroxide solution (4M, 784.06 uL, 5.60 eq) were added to a 100 mL three-necked flask with magnetic stirring. After the reaction system was purged with nitrogen for three times, Pd(PPh.sub.3).sub.2Cl.sub.2 (11.79 mg, 16.80 umol, 0.03 eq) was added. The reaction system was purged with nitrogen for three times again, then the reaction solution was stirred at 80 C. for 12 hours, and eventually turned from yellow to black. After completion of the reaction, water (30 mL) was added to the reaction solution and the mixture was extracted with dichloromethane (30 mL*3). The organic phase was combined, washed with saturated brine (30 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by column chromatography (PE:EA=1:0 to 20:1) to give the compound 79 (330.00 mg, crude). MS (ESI, M+1): 490.0.
(171) Step B: Compound 79 (330.00 mg, 672.89 umol, 1.00 eq) was dissolved in a mixed solvent of water (4.00 mL), methanol (8.00 mL) and tetrahydrofuran (8.00 mL). LiOH.H.sub.2O (141.17 mg, 3.36 mmol, 5.00 eq) was added to the above solution, which subsequently turned from yellow to light green. The reaction solution was stirred at 20 C. for 12 hours. After the completion of reaction, the reaction solution was evaporated to give a residue. Then 3 mol/L HCl (1.5 mL) was added to the residue to adjust the pH of the resulting solution to 5, the solution was filtered to give a filter cake, which was purified by preparative chromatography (formic acid) to give the product I-37 (33.70 mg, 71.43 umol, yield 10.62%, purity 98%).
(172) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.74 (s, 1H), 8.31 (br s, 1H), 7.62-7.49 (m, 2H), 7.44-7.35 (m, 3H), 7.34-7.27 (m, 2H), 7.23 (d, J=8.0 Hz, 1H), 7.10-7.03 (m, 1H), 7.02-6.93 (m, 3H), 6.57 (d, J=1.2 Hz, 1H), 6.40 (d, J=16.0 Hz, 1H), 2.75-2.58 (m, 2H), 0.99 (t, J=7.2 Hz, 3H); MS (ESI, M+1): 462.0.
Embodiment 38
(173) ##STR00120## ##STR00121##
(174) Step A: Compound 12 (150.00 mg, 318.20 umol, 1.00 eq), 2-meTHF (5.00 mL), 2-bromo-5-chloroanisole (84.57 mg, 381.84 umol, 1.20 eq), water (1.00 mL) and NaOH (50.91 mg, 1.27 mmol, 4.00 eq) were added successively to a single-neck flask with a magnetic stirrer. The resulting mixture was purged with N.sub.2 for three times. Then Pd(dppf)Cl.sub.2 (23.28 mg, 31.82 umol, 0.10 eq) was added to the mixture under nitrogen atmosphere. The reaction solution was stirred at 75 C. for 7 h. 10 mL EA and 10 mL water were added to the reaction solution. The organic phase was washed with 10 mL water, dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to give the crude product (162.00 mg), which was directly used in the next step base on the theoretical yield.
(175) Step B: Compound 80 (154.65 mg, 318.20 umol, 1.00 eq), THF (3.00 mL), MeOH (1.00 mL), water (1.00 mL) and LiOH (22.86 mg, 954.60 umol, 3.00 eq) were successively added to a single-neck flask with a magnetic stirrer. The resulting black solution was stirred at 45 C. for 4 h. 2 mL 1N HCl was added to the reaction solution, followed by addition of 10 mL water, then extracted with 30 mL EtOAc (10 mL*3). The organic phase was combined, dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to give a crude product, which was purified by preparative chromatography (formic acid) to give the compound 81 (28.00 mg, 61.14 umol, yield 19.21%).
(176) Compound 81 (28.00 mg, 61.14 umol, 1.00 eq) and CH.sub.3CN (3.00 mL) were successively added to a single-neck flask with a magnetic stirrer to give a yellow solution, followed by addition of NCS (8.16 mg, 61.14 umol, 1.00 eq). The resulting solution was stirred at 30 C. for 4 h. Then the reaction was quenched with 5 mL Na.sub.2S.sub.2O.sub.3 at 25 C., and 20 mL water was added. The mixture was extracted with 60 mL EtOAc (20 mL*3). The organic phase was combined, washed with 30 mL water, dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to give a crude product, which was purified by preparative chromatography (formic acid) to give I-38 (18.10 mg, 36.43 umol, yield 59.58%, purity 99.1%), MS (ESI, M+1): 492.1.
(177) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 11.44 (s, 1H), 7.50 (d, J=8.0 Hz, 1H), 7.41-7.36 (m, 4H), 7.20-7.10 (m, 3H), 6.95-6.88 (m, 4H), 6.37 (d, J=16.0 Hz, 1H), 3.82 (s, 3H), 2.01-1.98 (m, 2H), 086-0.83 (m, 3H).
Embodiment 39
(178) ##STR00122##
(179) Compound 12 (110.00 mg, 190.65 umol, 1.00 eq) and 2-trifluoromethyl-4-chlorobromobenzene (53.7 mg, 210 umol, 1.1 eq.) were dissolved in 4 mL dimethyltetrahydrofuran in a 100 mL single-neck flask, followed by addition of sodium hydroxide solution (4M, 142.99 uL, 3.00 eq) and Pd(PPh.sub.3).sub.2Cl.sub.2 (6.69 mg, 9.53 umol, 0.05 eq). The reaction system was purged with nitrogen for three times, and the reaction solution was stirred at 70 C. under nitrogen atmosphere for 16 hours. After completion of the reaction, the reaction solution was concentrated to give a residue, which was purified by preparative TLC (PE:EtOAc=0:1) to give a crude product. The crude product was further purified by preparative HPLC (FA) to give I-39.
(180) .sup.1H NMR (400 MHz, CHLOROFORM-d) =7.72 (s, 1H), 7.67 (d, J=7.8 Hz, 1H), 7.60 (d, J=15.8 Hz, 1H), 7.28-7.24 (m, 2H), 7.21-7.12 (m, 2H), 7.08 (d, J=2.6 Hz, 1H), 7.05-6.99 (m, 3H), 6.90 (dd, J=2.6, 8.7 Hz, 1H), 6.72 (d, J=1.5 Hz, 1H), 6.35 (d, J=15.9 Hz, 1H), 3.81 (s, 3H), 3.03 (m, J=7.5, 13.9 Hz, 1H), 2.65-2.57 (m, 1H), 1.08 (t, J=7.5 Hz, 3H); MS (ESI, M+1): 492.5.
Embodiment 40
(181) ##STR00123##
(182) Step A: Compound 13 (1 g, 2.11 mmol, 1 eq) and dichloromethane (20 mL) were added to a 100 mL single-neck flask with a magnetic stirrer. The solution was purged with nitrogen and cooled to 0 C., followed by addition of compound 82 (1.00 g, 3.16 mmol, 1.5 eq) at 0 C. The resulting dark green solution was stirred at 25 C. under nitrogen atmosphere for 16 hours. 15 mL saturated aqueous sodium bicarbonate solution containing 10% sodium thiosulfate was added, and the resulting mixture was stirred for 10 minutes. The aqueous phase was extracted three times with 30 mL dichloromethane each time. The organic phase was combined, washed with 20 mL saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography (EA/PE=0%-20%) to give the product 83 as a yellow solid.
(183) Step B: Compound 3 (1 g, (0.15 g, 304.91 umol, 1 eq), 2 mL ethanol and 3 mL tetrahydrofuran were added to a 100 mL single-neck flask with a magnetic stirrer, followed by addition of a solution of lithium hydroxide monohydrate (38.38 mg, 914.72 umol, 3 eq) in 1 mL water. The resulting yellow solution was stirred at 50 C. under nitrogen atmosphere for 2 hours, then concentrated and diluted with 10 mL water, adjusted pH to 5-6 with 1M hydrochloric acid, extracted three times with 20 mL ethyl acetate each time. The organic phase was combined, washed with 20 mL saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by preparative high performance liquid chromatography to give I-40. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.73 (s, 1H), 7.55 (d, J=8.0 Hz, 1H), 7.45-7.27 (m, 6H), 7.19-7.11 (m, 2H), 7.10-7.04 (m, 1H), 6.95 (d, J=8.4 Hz, 2H), 6.40 (d, J=16.0 Hz, 1H), 2.64-2.56 (m, 2H), 0.93 (t, J=7.6 Hz, 3H).
Embodiment 41
(184) ##STR00124##
(185) Step A: Compound 1 (400.00 mg, 693.27 umol, 1.00 eq), 2-chloro-4-methoxybromobenzene (230.32 mg, 1.04 mmol, 1.50 eq) and 10 mL 2-methyltetrahydrofuran were added to a 100 mL single-neck flask, followed by successive addition of aqueous potassium hydroxide solution (4M, 693.27 uL, 4.00 eq) and bis(triphenylphosphine)palladium dichloride (24.33 mg, 34.66 umol, 0.05 eq). The reaction solution was purged with nitrogen, then heated to 70 C. and stirred for 12 hours. The reaction solution turned from yellow to brown. After completion of the reaction, 20 mL ethyl acetate was added to the reaction solution, and the resulting mixture was filtered through celite. The filtrate was washed three times with 20 mL saturated brine each time, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product 84 as a brown jelly (570.00 mg, crude product), which was directly used in the next step.
(186) Step B: Lithium hydroxide monohydrate (490.93 mg, 11.70 mmol, 10.00 eq) was added to a solution of compound 84 (570.00 mg, 1.17 mmol, 1.00 eq) in a mixed solvent of 3 mL THF, 3 mL methanol and 3 mL water in a flask. The reaction solution was stirred at 45 C. for 1 hour and remained brown. After completion of the reaction, the reaction solution was adjusted to pH 1 with 1M hydrochloric acid, followed by addition of 20 mL ethyl acetate. The organic phase was washed twice with 20 mL saturated brine each time, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product, which was purified by preparative high performance liquid chromatography (formic acid system) to give the compound I-41. .sup.1H NMR (400 MHz, DMSO-d.sub.6): 10.72 (brs, 1H), 7.54 (d, J=7.6 Hz, 1H), 7.43-7.39 (m, 3H), 7.29 (d, J=8.0 Hz, 1H), 7.08-6.94 (m, 6H), 6.80 (dd, J=8.8 Hz, J=2.8 Hz, 1H), 6.54 (d, J=1.2 Hz, 1H), 6.39 (d, J=16.0 Hz, 1H), 3.71 (s, 3H), 2.66-2.55 (m, 2H), 0.98 (t, J=7.2 Hz, 3 H). MS (ESI, M+1): 458.1.
Embodiment 42
(187) ##STR00125## ##STR00126##
(188) Step A: A solution of NBS (42.85 mg, 240.74 umol, 0.90 eq) in dichloromethane (1 mL) was added to a solution of compound 84 (130.00 mg, 267.49 umol, 1.00 eq) in dichloromethane (4.00 mL) and acetonitrile (4.00 mL). The reaction solution was stirred for 1 hour, then quenched with aqueous sodium thiosulfate solution and extracted with dichloromethane. The organic phase was dried over sodium sulfate and concentrated to give the intermediate 85 (180 mg, crude product), which was directly used in the next step.
(189) Step B: A solution of intermediate 85 (180.00 mg, 318.64 umol, 1.00 eq) and cuprous cyanide (171.22 mg, 1.91 mmol, 417.61 uL, 6.00 eq) in N-methylpyrrolidone (2.00 mL) were heated to 180 C. under microwave irradiation and stirred for 2 hours. Then the reaction solution was cooled to room temperature, and additional cuprous cyanide (57.07 mg, 637.28 umol, 139.20 uL, 2.00 eq) was added, and the resulting reaction solution was heated to 180 C. under microwave irradiation and stirred for another 2 hours. The reaction solution was cooled to room temperature, poured into 20 mL water, extracted with ethyl acetate and concentrated to give a crude product, which was purified by preparative HPLC to give the intermediate 86 (30.00 mg, 54.60 umol, yield 17.13%, purity 93%).
(190) Step C: Lithium hydroxide (12.32 mg, 293.61 umol, 5.38 eq) was added to a solution of intermediate 86 (30.00 mg, 54.60 umol, 1.00 eq) in a mixed solvent of tetrahydrofuran (4.00 mL), methanol (1.00 mL) and water (1.00 mL). The reaction solution was stirred for 4 hours, then adjusted to pH 1 with 4M HCl, concentrated in vacuum and extracted with ethyl acetate. The organic phase was combined, dried over sodium sulfate and concentrated to give a crude product, which was purified by preparative HPLC to give I-42. .sup.1H NMR (400 MHz, DMSO-d.sub.6): 12.32 (s, 1H), 7.66 (d, J=8.8 Hz, 1H), 7.50 (d, J=8.0 Hz, 1H), 7.55-7.40 (m, 3H), 7.33-7.23 (m, 2H), 7.15 (d, J=8.8 Hz, 1H), 7.10-7.00 (m, 3H), 6.91 (dd, J=4.8 Hz, J=16 Hz, 1H), 6.42 (d, J=16 Hz, 1H), 3.75 st, 3H), 2.53-2.31 (m, 2H), 0.93 (t, J=7.2 Hz, 3 H). (ESI, M+1): 483.1.
Embodiment 43
(191) ##STR00127## ##STR00128##
(192) Step A: KOH (4M, 259.66 uL, 3.06 eq) and Pd(Ph.sub.3P).sub.2Cl.sub.2 (47.65 mg, 67.88 umol, 0.20 eq) were added to a solution of the intermediate 12 (200.00 mg, 339.42 umol, 1.00 eq) and 2-bromo-5-methoxyphenyl cyanide (108.00 mg, 509.13 umol, 1.50 eq) in 2-methyltetrahydrofuran. The reaction solution was stirred at 65 C. under nitrogen atmosphere for 13 hours, then cooled to room temperature, and additional Pd(Ph.sub.3P).sub.2Cl.sub.2 (20.00 mg, 33 umol, 0.10 eq) was added. The reaction solution was heated to 65 C. and stirred for another 3 hours to give 87, which was directly used in the next step. The reaction solution was cooled to room temperature, then methanol (6.00 mL) and water (3.00 mL) were added and the resulting mixture was stirred at 35 C. for 2 hours. Lithium hydroxide (15 mg) was added, the reaction solution was heated to 45 C. and stirred for 3 hours. The reaction solution was cooled to room temperature, adjusted to pH 5 with 2M HCl, diluted with water, extract with ethyl acetate, dried over sodium sulfate, concentrated and purified by silica gel column chromatography to give the intermediate 88.
(193) Step B: NCS (20.49 mg, 153.45 umol, 1.00 eq) was added to a solution of intermediate 88 (70.00 mg, 153.45 umol, 1.00 eq) in acetonitrile (3.00 mL). The reaction solution was stirred at 20 C. for 3 hours, then quenched with aqueous sodium thiosulfate solution (10 mL), adjusted to pH 5 with 2M HCl, extracted with dichloromethane. The organic phase was combined, dried over sodium sulfate and concentrated to give a crude product, which was purified by preparative HPLC to give the product I-43. .sup.1H NMR (400 MHz, DMSO-d.sub.6): 11.43 (s, 1H), 7.52 (d, J=8.4 Hz, 1H), 7.42-7.39 (m, 5H), 7.37-7.35 (m, 1H), 7.31 (d, J=2.4 Hz, 1H), 7.27-7.15 (m, 2H), 6.94 (d, J=8.4 Hz, 2H), 6.39 (d, J=16.0 Hz, 1H), 3.78 (s, 3H), 2.58-2.41 (m, 2H), 0.91 (t, J=7.6 Hz, 3H); MS (ESI, M+1): 483.1.
Embodiment 44
(194) ##STR00129##
(195) Step A: NCS (15.80 mg, 118.33 umol, 1.00 eq) was added to a solution of 48 (62.00 mg, 118.33 umol, 1.00 eq) in acetonitrile (3.00 mL) and dichloromethane (5.00 mL). The reaction solution was stirred at 15 C. for 5 hours. Additional NCS (4.00 mg, 29.96 umol, 0.25 eq) was added and the reaction solution was stirred for another 1 hour, then quenched with aqueous sodium thiosulfite solution (20 mL), extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over sodium sulfate to give the intermediate 89 (66.00 mg, crude product), which was used directly in the next step.
(196) Step B: LiOH (14.88 mg, 354.57 umol, 3.00 eq) was added to a solution of the intermediate 89 (66.00 mg, 118.19 umol, 1.00 eq) in a mixed solvent of methanol (3.00 mL), tetrahydrofuran (3.00 mL) and water (3.00 mL). The reaction solution was stirred at 15 C. for 12 hours and concentrated. The residue was diluted with water (10 mL), adjusted to pH 5 with 2M HCl, extracted with dichloromethane. The organic phase was combined, washed with saturated brine, dried over sodium sulfate and concentrated to give a crude product, which was purified by preparative HPLC to give I-44. .sup.1H NMR (400 MHz, DMSO-d.sub.6): 11.51 (s, 1H), 7.81 (d, J=2.0 Hz, 1H), 7.73 (d, J=12 Hz, 1H), 7.49 (d, J=9.6 Hz, 1H), 7.46-7.35 (m, 5H), 7.22-7.13 (m, 2H), 6.97 (d, J=8.4 Hz, 2H), 6.39 (s, J=16.0 Hz, 1H), 2.55-2.47 (m, 1H), 2.38-2.25 (m, 1H), 0.87 (t, J=7.2 Hz, 3H); MS (ESI, M+1): 530.1.
Embodiment 45
(197) ##STR00130##
(198) Step A: NCS (9.48 mg, 71.00 umol, 0.9 eq) was added to a mixture of compound II-1 (0.04 g, 78.89 umol, 1 eq) and dichloromethane (4 mL) in a single-neck flask with a magnetic stirrer. The yellow solution was stirred at 15 C. for 2 hours, then concentrated to give a crude product, which was purified by preparative high performance liquid chromatography to give II-5.
Embodiment 46
(199) ##STR00131##
(200) Step A: Aqueous sodium hydroxide solution (4M), 1.00 mL, 4.00 eq) was added to a mixture of compound 11 (423.16 mg, 1.00 mmol, 1.00 eq) (a solution of 10 mL 2-methyltetrahydrofuran) and compound 29 (402.19 mg, 1.20 mmol, 1.20 eq) in a three-necked flask, followed by addition of bis(diphenylphosphino)ferrocene dichloropalladium (36.50 mg, 50.00 umol, 0.05 eq) under nitrogen atmosphere. The reaction solution was stirred at 60 C. for 4 hours. After completion of the reaction, the reaction solution was directly used in the next step without further posttreatment based on the theoretical yield for 90.
(201) Step B: Dichlorobis(tripheny/phosphine)palladium(II) (42.12 mg, 60.00 umol, 4.00 eq) and 1-chloro-2-iodobenzene (291.14 mg, 1.20 mmol, 1.20 eq) under nitrogen atmosphere in a three-necked flask. The reaction solution was added at 70 C. under nitrogen atmosphere for 4 hours. After completion of the reaction, water (40 mL) and ethyl acetate (30 mL) were added to the reaction solution, then extracted with ethyl acetate (20 mL*3). The organic phase was combined, washed with water (30 mL), dried over sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative liquid chromatography (prep. HPLC: formic acid) to give II-6.
(202) .sup.1H NMR (400 MHz, DMSO-d.sub.6) =10.65 (s, 1H), 8.35 (s, 1H), 7.54-6.84 (m, 10H), 6.62-6.54 (m, 3H), 4.493 (dd, J=6.4 Hz, 48 Hz, 2H), 3.78 (d, J=5.2 Hz, 2H), 2.95 (t, J=6.4 Hz, 2H), 2.68-2.51 (m, 6H), 0.99 (t, J=7.6 Hz, 3H); MS (ESI, M+1): 489.1.
Embodiment 47
(203) ##STR00132##
(204) Step A: 2-Iodobenzonitrile (329.79 mg, 1.44 mmol, 1.20 eq) was added to a mixture of compound 90 (605.33 mg, 1.20 mmol, 1.00 eq) (a solution in 10 mL 2-methyltetrahydrofuran), water (2.00 mL) and sodium hydroxide (192.00 mg, 4.80 mmol, 4.00 eq) in a single-neck flask, which was then purged with N.sub.2 for three times. Bis(triphenylphosphine) palladium chloride (42.11 mg, 60.00 umol, 0.05 eq) was added under nitrogen atmosphere. The reaction solution was stirred at 70 C. for 7 hours. After completion of the reaction, the reaction solution was diluted with 40 mL water, extracted with 45 mL ethyl acetate (15 mL*3). The organic phase was combined, washed with 40 mL water, dried over Na.sub.2SO.sub.4, filtered and concentrated to give a crude product, which was purified by preparative chromatography (formic acid system) to give the compound II-7. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.65 (s, 1H), 8.27 (s, 1H), 7.66-7.51 (m, 4H), 7.38-7.30 (m, 2H), 7.06-7.00 (m, 2H), 6.81 (d, J=8.8 Hz, 2H), 6.62 (d, J=8.8 Hz, 2H), 6.57 (d, J=1.6 Hz, 1H), 4.54 (d, J=6.0 Hz, 1H), 4.42 (d, J=6.0 Hz, 1H), 3.78 (t, J=5.6 Hz, 2H), 2.96 (t, J=5.6 Hz, 2H), 2.68-2.63 (m, 6H), 2.03-1.96 (m, 1H), 1.01 (t, J=7.6 Hz, 3H); MS (ESI, M+1): 480.3.
Embodiment 48
(205) ##STR00133## ##STR00134##
(206) Step A: Diboron pinacol ester (1.16 g, 4.57 mmol, 1.05 eq) and tetrakis(triphenylphosphine)platinum (271.04 mg, 217.84 umol, 0.05 eq) were added to a solution of compound 10 (750.00 mg, 4.36 mmol, 1.00 eq) in 2-methyltetrahydrofuran (20.00 mL), the reaction solution was purged with nitrogen for three times, then stirred at 70 C. (internal temperature 60 C.) under nitrogen atmosphere for 5 hours, and the resulting 11 was not isolated. After the reaction solution was cooled to 0 C., compound 91 (1.34 g, 3.91 mmol, 0.90 eq), potassium carbonate (1.20 g, 8.70 mmol, 2.00 eq), 6 mL water, 30 mL 2-methyltetrahydrofuran and dichlorobis(triphenylphosphine)palladium (152.60 mg, 217.41 umol, 0.05 eq) were added, the resulting mixture was purged with nitrogen for three times and stirred at 30 C. under nitrogen atmosphere for 12 hours, and the resulting 92 was not isolated. Then, 4-fluoro-2-chloroiodobenzene (556.54 mg, 2.17 mmol, 2.00 eq), aqueous potassium hydroxide solution (4M, 1.09 mL, 4.00 eq), dichlorobis(triphenylphosphine)palladium (76.17 mg, 108.51 umol, 0.10 eq) and 9 mL 2-methyltetrahydrofuran were added, and the resulting mixture was purged with nitrogen for three times and stirred at 60 C. under nitrogen atmosphere for 12 hours. Then 20 mL ethyl acetate was added, and the mixture was filtered. the filtrate was washed three times with 20 mL saturated brine each time, dried over anhydrous sodium sulfate, filtered, concentrated and purified by preparative high performance liquid chromatography (formic acid system) to give the compound 93 as a yellow solid (41.10 mg, 71.26 umol, yield 6.54%, purity 97.1%). MS (ESI, M+23): 536.1.
(207) Step B: N-chlorosuccinimide was added to a solution of compound 93 (41.10 mg, 77.64 umol, 1.00 eq) in 10 mL acetonitrile, and the reaction solution was stirred at 30 C. for 10 hours. After completion of the reaction, the reaction solution was directly concentrated to give a crude product, which was purified by preparative high performance liquid chromatography (formic acid system) to give II-8. .sup.1H NMR (400 MHz, DMSO-d.sub.6): 11.42 (s, 1H), 8.36 (s, 1H), 7.49 (d, J=7.6 Hz, 1H), 7.42-7.37 (m, 2H), 7.26 (dd, J=8.8 Hz, J=6.4 Hz, 1H), 7.19-7.10 (m, 3H), 6.92 (d, J=8.4 Hz, 2H), 6.64 (d, J=8.8 Hz, 2H), 3.77 (t, J=5.6 Hz, 2H), 3.33 (t, J=7.2 Hz, 2H), 2.91 (t, J=7.6 Hz, 2H), 2.71 (d, J=6.8 Hz, 2H), 2.65-2.59 (m, 3H), 2.43-2.37 (m, 2H), 0.92 (t, J=7.6 Hz, 3 H). MS (ESI, M+1): 548.1.
Embodiment 49
(208) ##STR00135##
(209) Step A: Diboron pinacol ester (147.80 mg, 582.03 umol, 1.05 eq) and tetrakis(triphenylphosphine)platinum (34.48 mg, 27.72 umol, 0.05 eq) were added to a solution of compound 10 (100.00 mg, 554.31 umol, 1.00 eq) in 2-methyltetrahydrofuran (10.00 mL), the reaction solution was purged with nitrogen for three times, then stirred at 70 C. under nitrogen atmosphere for 5 hours, and the resulting 11 was not isolated. After the reaction solution was cooled to 0 C., compound 91 (238.50 mg, 497.68 umol, 0.90 eq), potassium carbonate K.sub.2CO.sub.3 (152.91 mg, 1.11 mmol, 2.00 eq), 2.5 mL water, 10 mL 2-methyltetrahydrofuran and dichlorobis(triphenylphosphine)palladium (19.41 mg, 27.65 umol, 0.05 eq) were added, the resulting mixture was purged with nitrogen for three times and stirred at 30 C. under nitrogen atmosphere for 12 hours, and the resulting 92 was not isolated. Then, 4-chloro-2-trifluoromethyl iodobenzene (338.89 mg, 1.11 mmol, 2.00 eq), aqueous potassium hydroxide solution (4M, 552.93 uL, 4.00 eq), dichlorobis(triphenylphosphine)palladium (19.40 mg, 27.65 umol, 0.05 eq) and 10 mL 2-methyltetrahydrofuran were added, and the resulting mixture was purged with nitrogen for three times and stirred at 70 C. under nitrogen atmosphere for 12 hours. Then 20 mL ethyl acetate was added, and the mixture was filtered. The filtrate was washed three times with 20 mL saturated brine each time, dried over anhydrous sodium sulfate, filtered, concentrated and purified by preparative high performance liquid chromatography (formic acid system) to give the compound II-9. .sup.1H NMR (400 MHz, DMSO-d.sub.6): 10.67 (s, 1H), 8.38 (s, 1H), 7.71 (d, J=1.6 Hz, 1H), 7.63 (dd, J=8.4 Hz, J=1.6 Hz, 1H), 7.53 (d, J=8.0 Hz, 1H), 7.32 (dd, J=8.0 Hz, J=12.0 Hz, 2H), 7.04 (t, J=7.2 Hz, 1H), 6.98 (t, J=7.2 Hz, 1H), 6.80 (d, J=8.8 Hz, 2H), 6.63 (d, J=8.4 Hz, 2H), 6.48 (d, J=0.8 Hz, 1H), 3.77 (t, J=5.6 Hz, 2H), 3.33 (t, J=7.2 Hz, 2H), 2.92 (t, J=7.6 Hz, 2H), 2.84-2.77 (m, 1H), 2.71 (d, J=6.8 Hz, 2H), 2.66-2.61 (m, 3H), 2.45-2.38 (m, 1H), 0.93 (t, J=7.6 Hz, 3 H). MS (ESI, M+23): 564.2.
Embodiment 50
(210) ##STR00136##
(211) Step A: 1-Chloro-2-iodobenzene (517.50 mg, 2.17 mmol, 2.00 eq), potassium aqueous hydroxide (4M, 1.09 mL, 4.00 eq) and 2-methyltetrahydrofuran (5.00 mL) was added to a solution of compound 92 (555.00 mg, 1.09 mmol, 1.00 eq) in 2-methyltetrahydrofuran (7.50 mL), the reaction system was purged with nitrogen for three times, then dichlorobis(triphenylphosphine)palladium (38.08 mg, 54.26 umol, 0.05 eq) was added. The reaction solution was stirred at 70 C. under nitrogen atmosphere for 12 hours, then diluted with 20 mL ethyl acetate and filtered. The filtrate was washed three times with 20 mL brine each time, dried over anhydrous sodium sulfate, filtered, concentrated and purified by preparative HPLC (formic acid system) to give the compound II-10. .sup.1H NMR (400 MHz, DMSO-d.sub.6): 10.64 (brs, 1H), 8.29 (brs, 1H), 7.54 (d, J=8.0 Hz, 1H), 7.37-7.34 (m, 1H), 7.29 (d, J=8.0 Hz, 1H), 7.21-7.18 (m, 2H), 7.14-7.12 (m, 1H), 7.04 (dt, J=7.2 Hz, J=1.2 Hz, 1H), 6.97 (dt, J=8.0 Hz, J=1.2 Hz, 1H), 6.90-6.82 (m, 2H), 6.61-6.59 (m, 2H), 6.53 (d, J=1.2 Hz, 1H), 3.76 (t, J=5.6 Hz, 2 H), 3.33 (t, J=7.2 Hz, 2 H), 2.92 (t, J=7.2 Hz, 2 H), 2.71 (d, J=6.8 Hz, 2 H), 2.67-2.55 (m, 5H), 0.97 (t, J=7.6 Hz, 3 H). MS (ESI, M+1): 496.2.
Embodiment 51
(212) ##STR00137##
(213) Step A: Diboron pinacol ester (147.80 mg, 582.03 umol, 1.05 eq) and tetrakis(triphenylphosphine)platinum (34.48 mg, 27.72 umol, 0.05 eq) were added to a solution of compound 10 (100.00 mg, 554.31 umol, 1.00 eq) in 2-methyltetrahydrofuran (10.00 mL), the reaction solution was purged with nitrogen for three times, then stirred at 70 C. under nitrogen atmosphere for 5 hours, and the resulting 11 was not isolated. After the reaction solution was cooled to 0 C., compound 91 (238.50 mg, 497.68 umol, 0.90 eq), potassium carbonate K.sub.2CO.sub.3 (152.91 mg, 1.11 mmol, 2.00 eq), 2.5 mL water, 10 mL 2-methyltetrahydrofuran and dichlorobis(triphenylphosphine)palladium (19.41 mg, 27.65 umol, 0.05 eq) were added, the resulting mixture was purged with nitrogen for three times and stirred at 30 C. under nitrogen atmosphere for 12 hours, and the resulting 92 was not isolated. Then, 2-iodobenzonitrile (253.26 mg, 1.11 mmol, 2.00 eq), aqueous potassium hydroxide solution (4M, 552.93 uL, 4.00 eq), dichlorobis(triphenylphosphine)palladium (19.40 mg, 27.65 umol, 0.05 eq) and 5 mL 2-methyltetrahydrofuran were added, and the resulting mixture was purged with nitrogen for three times and stirred at 70 C. under nitrogen atmosphere for 12 hours. Then 20 mL ethyl acetate was added, and the mixture was filtered. The filtrate was washed three times with 20 mL saturated brine each time, dried over anhydrous sodium sulfate, filtered, concentrated and purified by preparative high performance liquid chromatography (formic acid system) to give the compound II-11. .sup.1H NMR (400 MHz, DMSO-d.sub.6): 10.65 (s, 1H), 8.28 (s, 1H), 7.65-7.61 (m, 2H), 7.56 (d, J=7.6 Hz, 1H), 7.50 (d, J=7.6 Hz, 1H), 7.35 (dt, J=7.6 Hz, J=0.8 Hz, 1H), 7.30 (d, J=8.0 Hz, 1H), 7.05 (dt, J=8.0 Hz, J=1.2 Hz, 1H), 6.99 (dt, J=8.0 Hz, J=1.2 Hz, 1H), 6.80 (d, J=8.8 Hz, 2H), 6.61 (d, J=8.8 Hz, 2H), 6.56 (d, J=1.6 Hz, 1H), 3.77 (t, J=5.2 Hz, 2H), 3.34 (t, J=7.2 Hz, 2H), 2.92 (t, J=6.8 Hz, 2H), 2.71 (d, J=7.2 Hz, 2H), 2.67-2.57 (m, 3H), 2.52-2.50 (m, 2H), 1.00 (t, J=7.6 Hz, 3 H); MS (ESI, M+1, M+23): 492.2, 509.1.
Embodiment 52
(214) ##STR00138##
(215) A solution of 51 (100.00 mg, 218.58 umol, 1.00 eq), 2,4-difluoroiodobenzene (73.44 mg, 306.01 umol, 1.40 eq) and bis(diphenylphosphino)ferrocene dichloropalladium (7.94 mg, 10.86 umol, 0.05 eq) in 2-methyltetrahydrofuran (15 mL) was purged with nitrogen, followed by addition of aqueous sodium hydroxide (4M, 218.58 uL, 4.00 eq). The reaction solution was stirred at 65 C. for 12 hours and turned from yellow to black. The resulting 15 mL black solution containing compound 94, the theoretical amount of which was 150 mg, was directly used in the next step. The above 15 mL black solution containing compound 94, the theoretical amount of which was 150 mg, and methanol (8.00 mL) were stirred at 30 C. for 15 hours. 20 mL was added to the reaction solution, then the mixture was filtered. The filtrate was adjusted to pH 4 with 3M hydrochloric acid, then extracted three times with 20 mL ethyl acetate. The organic phase was combined, washed once with 20 mL saturated brine, filtered, dried over anhydrous sodium sulfate and concentrated to give a crude product, which was dissolved in 3 mL acetonitrile and purified by high performance liquid chromatography (formic acid system) to give I-45. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.76 (s, 1H), 7.55 (d, J=7.6 Hz, 1H), 7.47-7.38 (m, 3H), 7.33-7.19 (m, 2H), 7.15-6.91 (m, 6H), 6.54 (d, J=1.6 Hz, 1H), 6.41 (d, J=16.4 Hz, 1H), 2.63 (q, J=7.2 Hz, 2H), 0.99 (t, J=7.6 Hz, 3H). MS (ESI, M+1): 430.1.
Embodiment 53
(216) ##STR00139## ##STR00140##
(217) Step A: A solution of 2-chloro-4-fluorobenzaldehyde (5.00 g, 31.53 mmol, 1.00 eq) in 12 mL tetrahydrofuran was cooled to 20 C., followed by addition of ethylmagnesium bromide (2M, 17.34 mL, 1.10 eq) under nitrogen atmosphere. The reaction solution was warmed to 0 C. and then stirred at 20 C. for 12 hours. The reaction solution was quenched with saturated 15 mL aqueous ammonium chloride solution at 0 C., diluted with 10 mL water, partitioned, the aqueous phase was extracted three times with 50 mL ethyl acetate each time. The organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by silica gel column chromatography (EA/PE=0% to 30%) to give the product 101 as a yellow oil.
(218) Step B: 101 (1.70 g, 9.01 mmol, 1.00 eq) and 50 mL dichloromethane were added to a single-neck round bottom flask, then cooled to 0 C. and Dess Martin reagent (6.11 g, 14.42 mmol, 4.46 mL, 1.60 eq) was added under nitrogen atmosphere. The reaction solution was stirred at 0 C. for 1 hour, then quenched with saturated aqueous sodium bicarbonate solution containing 15% sodium thiosulfate (20 mL) and filtered. The filtrate was partitioned, and the aqueous phase was extracted three times with 50 mL dichloromethane each time. The organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography (EA/PE=0% to 10%) to give 98.
(219) Step C: 5-Azaindole (10.00 g, 84.65 mmol, 1.00 eq), Boc.sub.2O (19.40 g, 88.88 mmol, 20.42 mL, 1.05 eq) and 200 mL dichloromethane were added to a single-neck round bottom flask, followed by addition of DMAP (1.03 g, 8.47 mmol, 0.10 eq) under nitrogen atmosphere. The reaction solution was stirred at 20 C. for 12 hours, then concentrated and purified by silica gel column chromatography (EA/PE=0%-40%) to give 95. .sup.1H NMR (400 MHz, CHLOROFORM-d) 8.89 (s, 1H), 8.47 (d, J=6.0 Hz, 1H), 7.98 (d, J=5.6 Hz, 1H), 7.61 (d, J=3.6 Hz, 1H), 6.65 (d, J=3.6 Hz, 1H), 1.69 (s, 9H).
(220) Step D: 95 (1.00 g, 4.58 mmol, 1.00 eq) was dissolved in 10 mL tetrahydrofuran and cooled to 78 C. LDA (2M, 2.75 mL, 1.20 eq) was added dropwise, and the mixture was maintained below 70 C. for 30 minutes, then a solution of 4-iodobenzaldehyde (1.06 g, 4.58 mmol, 1.00 eq) in 5 mL tetrahydrofuran was added dropwise at 70 C. and the reaction solution was stirred at this temperature for 1 hour. The reaction solution was slowly quenched with 10 mL saturated aqueous ammonium chloride solution, then diluted with 10 mL water, partitioned, and the aqueous phase was extracted three times with 30 mL ethyl acetate each time. The organic phase was combined, washed with 20 mL saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by silica gel column chromatography (EA/PE=20%-100%) to give the product 96 as a yellow solid. MS (ESI, M+1): 450.9.
(221) Step E: 96 (6.10 g, 13.55 mmol, 1.00 eq) and 100 mL tetrahydrofuran were added to a single-neck round bottom flask, followed by addition of manganese dioxide (11.78 g, 135.50 mmol, 10.00 eq). The reaction solution was stirred at 70 C. for 72 hours, then filtered. The filter cake was washed with 30 mL dichloromethane. The filtrate was concentrated and purified by silica gel column chromatography (methanol/dichloromethane=0%-10%), and further purified by preparative high performance liquid chromatography (basic) to give the product 97 as a yellow solid. MS (ESI, M+1): 349.0.
(222) Step D: Zinc powder (657.39 mg, 10.05 mmol, 14.00 eq) was added to 5 mL tetrahydrofuran, followed by dropwise addition of titanium tetrachloride (817.26 mg, 4.31 mmol, 472.40 uL, 6.00 eq) at 30 C. Then a solution of 97 (250.00 mg, 718.10 umol, 1.00 eq) and 98 (415.41 mg, 2.23 mmol, 3.10 eq) in tetrahydrofuran (20.00 mL) was added. The reaction solution was stirred at 70 C. for 14 hours and turned bright yellow. The reaction solution was cooled to room temperature, quenched with 20 mL saturated aqueous sodium bicarbonate solution, filtered to remove the yellow solid and partitioned. The aqueous phase was extracted three times with 20 mL ethyl acetate each time. The organic phase was combined, washed with brine, dried over anhydrous sodium sulfate, filtered. The filtrate was filtered, concentrated and purified by silica gel column chromatography (methanol/dichloromethane=0%-5%) to give the product 99. MS (ESI, M+1): 503.0.
(223) Step F: 99 (230.00 mg, 457.48 umol, 1.00 eq), methyl acrylate (196.92 mg, 2.29 mmol, 205.13 uL, 5.00 eq), 1 mL triethylamine, POT (69.62 mg, 228.74 umol, 0.50 eq) and 3 mL DMF were added to a three-necked round bottom flask, followed by addition of palladium acetate (30.81 mg, 137.24 umol, 0.30 eq) under nitrogen atmosphere. The reaction solution was purged with nitrogen, and stirred at 110 C. for 2 hours. Then the reaction solution was filtered, and the filter cake was washed with 20 mL dichloromethane. The filtrate was concentrated to give 100 (250.00 mg, crude). MS (ESI, M+1): 461.1.
(224) Step F: Lithium hydroxide monohydrate (57.35 mg, 1.37 mmol, 3.00 eq) was added to a solution of 100 (210.00 mg, 455.60 umol, 1.00 eq) in a mixed solvent of 5 mL tetrahydrofuran, 5 mL methanol and 5 mL water. The yellow suspension was stirred at 20 C. for 36 hours, then concentrated under reduced pressure, diluted with 10 mL water, and adjusted to pH 4-5 with citric acid. A solid precipitated, which was collected and purified by preparative high performance liquid chromatography (formic acid) to give the product I-46. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =12.75 (s, 1H), 9.31 (s, 1H), 8.43 (d, J=6.8 Hz, 1H), 7.86 (d, J=6.4 Hz, 1H), 7.51-7.43 (m, 3H), 7.39 (dd, J=2.4, 8.8 Hz, 1H), 7.32 (dd, J=6.4, 8.4 Hz, 1H), 7.20-7.13 (m, 2H), 7.01 (d, J=8.0 Hz, 2H), 6.45 (d, J=16.0 Hz, 1H), 2.61 (q, J=7.2 Hz, 2H), 1.00 (br t, J=7.6 Hz, 3H); MS (ESI, M+1): 447.0.
Embodiment 54
(225) ##STR00141##
(226) Step A: DMF (5.00 mL) was added a 50 mL single-neck flask, followed by addition of compound 48 (100.00 mg, 190.85 umol, 1.00 eq) and formamide (51.58 mg, 1.15 mmol, 45.64 uL, 6.00 eq). A solution of sodium methoxide in methanol (0.5M, 1.15 mL, 3.00 eq) was added to the solution. The reaction solution was stirred at 100 C. for 2 hours and turned from yellow to brown. After completion of the reaction, 30 mL ethyl acetate was added, then washed three times with 30 mL saturated brine each time. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated and purified by preparative high performance liquid chromatography (formic acid system) to give the product I-47. .sup.1H NMR (400 MHz, DMSO-d.sub.6): 10.76 (brs, 1H), 7.72 (d, J=2.0 Hz, 1H), 7.65 (dd, J=8.4 Hz, J=2.0 Hz, 1H), 7.55 (d, J=7.6 Hz, 1H), 7.44 (brs, 1H), 7.38 (d, J=8.4 Hz, 1H), 7.32-7.27 (m, 3H), 7.25 (d, J=15.6 Hz, 1H), 7.08-6.97 (m, 3H), 6.93 (d, J=8.0 Hz, 2H), 6.52 (d, J=1.6 Hz, 1H), 6.47 (d, J=16.0 Hz, 1H), 2.85-2.80 (m, 1H), 2.47-2.44 (m, 1H), 0.94 (t, J=7.6 Hz, 3 H). MS (ESI, M+1): 495.2.
Embodiment 55
(227) ##STR00142##
(228) Step A: Compound I-14 (100.00 mg, 177.45 umol, 1.00 eq), EDCI (51.03 mg, 266.17 umol, 1.50 eq) and DMAP (49.86 mg, 408.13 umol, 2.30 eq) were dissolved in dichloromethane (7.00 mL), followed by addition of methylsulfonamide (50.64 mg, 532.34 umol, 3.00 eq). The black reaction solution was stirred at 30 C. for 12 hours. After completion of the reaction, water (10 mL) was added, then extracted with ethyl acetate (10 mL*3). The organic phase was combined, washed once with 15 mL brine, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative chromatography (formic acid) to give the product I-48. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.80 (s, 1H), 8.15 (s, 1H), 7.75 (d, J=2.4 Hz, 1H), 7.66 (dd, J=2.0, 8.4 Hz, 1H), 7.57 (d, J=8.0 Hz, 1H), 7.45-7.29 (m, 5H), 7.11-6.91 (m, 4H), 6.54 (d, J=2.0 Hz, 1H), 6.46 (d, J=15.4 Hz, 1H), 3.13 (s, 3H), 2.89-2.78 (m, 1H), 2.46 (br d, J=7.6 Hz, 1H), 0.95 (t, J=7.6 Hz, 3H); MS (ESI, M+1): 573.1.
Embodiment 56
(229) ##STR00143##
(230) Step A: Compound 12 (272.73 mg, 509.13 umol, 1.00 eq), 2-cyano bromobenzene (139.01 mg, 763.70 umol, 1.50 eq), dichlorobis(triphenylphosphine)palladium (107.21 mg, 152.74 umol, 0.30 eq), aqueous potassium hydroxide (4M, 712.78 uL, 5.60 eq) were dissolved in 20 mL 2-methyltetrahydrofuran, the reaction solution was purged with nitrogen for three times, then stirred at 75 C. under nitrogen atmosphere for 7 hours. The reaction solution turned from yellow to dark brown and a black solid precipitated. The reaction solution was filtered through celite, and the filtrate was concentrated and purified to give the crude product 102 as a dark brown oil (250.00 mg, crude product), which was directly used in the next step without further purification.
(231) Step B: Compound 102 (250.00 mg, 559.86 umol, 1.00 eq), lithium hydroxide (67.04 mg, 2.80 mmol, 5.00 eq) were dissolved in a mixed solvent of 10 mL methanol, 5 mL tetrahydrofuran and 5 mL water. The reaction solution was stirred at 30 C. for 1 hour, then concentrated to 1 ml, adjusted to pH 5-6 with hydrochloric acid (3 mol/L), extracted twice with 100 mL dichloromethane (50 mL each time). The organic phase was combined, dried, filtered, concentrated under reduced pressure, and purified by preparative chromatography (formic acid system) to give the product I-49 as a yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.76 (s, 1H), 7.69-7.62 (m, 2H), 7.60-7.55 (m, 2H), 7.44-7.39 (m, 3H), 7.38-7.29 (m, 2H), 7.07 (t, J=7.6 Hz, 1H), 7.03-6.98 (m, 1H), 6.94 (d, J=8.0 Hz, 2H), 6.61 (s, 1H), 6.41 (d, J=16.0 Hz, 1H), 2.81-2.67 (m, 2H), 1.02 (br t, J=7.2 Hz, 3H); MS [ESI, M+1]:419.2.
Embodiment 57
(232) ##STR00144##
(233) I-50 was prepared using the intermediate 12 (227.27 mg, 424.27 mg, 1.00 eq) and o-methyliodobenzene (185.01 mg, 848.54 umol, 108.19 uL, 2.00 eq) as raw material through a similar procedure to that described in preparing I-49.
(234) The crude product was purified by preparative high performance liquid chromatography (formic acid system) to give the product I-50.
(235) .sup.1H NMR (400 MHz, DMSO-d.sub.6): 10.81 (brs, 1H), 7.54 (d, J=8.0 Hz, 1H), 7.39 (d, J=16.0 Hz, 1H), 7.34 (d, J=8.4 Hz, 2H), 7.30 (dd, J=8.0 Hz, J=0.8 Hz, 1H), 7.18 (dt, J=7.6 Hz, J=1.6 Hz, 1H), 7.14-7.03 (m, 4H), 6.99 (dt, J=7.2 Hz, J=1.2 Hz, 1H), 6.89 (d, J=8.4 Hz, 2H), 6.53 (d, J=1.2 Hz, 1H), 6.36 (d, J=16.0 Hz, 1H), 2.59-2.54 (m, 2H), 2.08 (s, 3H), 0.95 (t, J=7.6 Hz, 3 H). MS (ESI, M+1): 408.1.
Embodiment 58
(236) ##STR00145##
(237) I-51 was prepared using the intermediate 12 (327.27 mg, 610.96 umol, 1.00 eq) and o-chloroiodobenzene (291.37 mg, 1.22 mmol, 2.00 eq) as raw material through a similar procedure to that described in preparing I-49.
(238) The crude product was purified by preparative chromatography (formic acid system) to give the product I-51. MS [ESI, M+1].sup.+: 428.1, .sup.1H NMR (400 MHz, DMSO-d.sub.6) =10.74 (s, 1H), 7.56 (d, J=8.0 Hz, 1H), 7.42-7.35 (m, 4H), 7.30 (d, J=8.2 Hz, 1H), 7.24-7.15 (m, 3H), 7.09-6.94 (m, 4H), 6.57 (d, J=1.2 Hz, 1H), 6.38 (d, J=16.0 Hz, 1H), 2.75-2.59 (m, 1H), 2.75-2.56 (m, 1H), 1.00 (t, J=7.5 Hz, 3H).
Embodiment 59
(239) ##STR00146##
(240) I-52 was prepared using the intermediate 51 (101.68 mg, 221.57 umol, 1.00 eq) and 2-cyano-3-fluoroiodobenzene (65.68 mg, 265.88 umol, 1.20 eq) as raw material through a similar procedure to that described in preparing I-28.
(241) The crude product was purified by preparative high performance liquid chromatography (formic acid system) to give the product I-52. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =10.79 (s, 1H), 8.34 (br s, 1H), 7.79-7.68 (m, 1H), 7.60 (d, J=7.6 Hz, 1H), 7.47-7.39 (m, 3H), 7.34 (dd, J=8.0, 16.8 Hz, 3H), 7.11-7.06 (m, 1H), 7.04-6.99 (m, 1H), 6.95 (d, J=8.0 Hz, 2H), 6.62 (d, J=1.2 Hz, 1H), 6.43 (d, J=16.0 Hz, 1H), 2.89-2.68 (m, 2H), 1.05 (t, J=7.4 Hz, 3H). MS (ESI, M+1): 437.1.
Embodiment 60
(242) ##STR00147## ##STR00148##
(243) Step A: Bis(triphenylphosphine) palladium dichloride (30.00 mg, 42.74 umol, 0.5 eq) was added a solution of compound 12 (500.00 mg, 848.55 umol, 1.00 eq), 2,4-dichloroiodobenzene (250.00 mg, 916.43 umol, 1.08 eq), potassium hydroxide (450.00 mg, 8.02 mmol, 9.45 eq) in 2-methyltetrahydrofuran (8.00 mL) at 25 C. under nitrogen atmosphere. The reaction solution was stirred at 70 C. for 18 hours, then filtered, extracted, and purified by silica gel column chromatography to give 103.
(244) Step B: NBS (115.00 mg, 646.14 umol, 1.02 eq) was added to a solution of compound 103 (330.00 mg, 632.52 umol, 1.00 eq) in 5 mL acetonitrile. The reaction solution was stirred at 25 C. for 10 minutes, then 5 mL dichloromethane was added and the reaction solution was stirred at 25 C. for another 50 minutes. The reaction solution was extracted, partitioned, dried over anhydrous sodium sulfate and concentrated to give the product 104 as a yellow solid.
(245) Step C: Cuprous cyanide (100.00 mg, 1.1.2 mmol, 243.90 uL, 21.46 eq) was added to a solution of compound 104 (125.00 mg, 52.04 umol, 1.00 eq) in 5 mL NMP. The reaction solution was stirred at 180 C. under microwave irradiation for 3 hours. Then the reaction solution was adjusted to pH 9-10 with 20 mL aqueous sodium bicarbonate, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate and concentrated to give 105.
(246) Step D: Lithium hydroxide monohydrate (75.00 mg, 1.79 mmol, 10.24 eq) was added to a solution of compound 105 (90.00 mg, 174.61 umol, 1.00 eq) in a mixed solvent of 2 mL THF, 0.5 mL water and 2 mL methanol. The reaction solution was stirred at 25 C. for 9 hours. 10 mL water was added to the reaction solution, then the resulting solution was adjusted to pH 3-4 with 1M hydrochloric acid, extracted with ethyl acetate, concentrated to give a crude product, which was purified by preparative high performance liquid chromatography to give I-53. .sup.1H NMR (400 MHz, CDCl3): 8.50 (s, 1H), 7.80-7.34 (m, 5H), 7.26-7.75 (m, 1H), 7.64 (d, J=16.0 Hz, 1H), 7.42-7.27 (m, 5H), 7.16 (dd, J=2.0 Hz, J=9.6 Hz, 1H), 7.08 (t, J=4.8 Hz, 3 H), 6.31 (d, J=16.0 Hz, 1 H), 2.78-2.65 (m, 1H), 7.26-7.75 (m, 1H), 7.63-2.51 (m, 1H), 1.05 (t, J=7.6 Hz, 3 H); MS (ESI, M+1): 487.2.
Embodiment 61
(247) ##STR00149## ##STR00150##
(248) Step A: Intermediate 106 was prepared using intermediate 51 (200.00 mg, 443.13 umol, 1.00 eq) and m-methyliodobenzene (135.26 mg, 620.38 umol, 79.56 uL, 1.40 eq) as raw material through a similar procedure to that described in preparing intermediate 54.
(249) Step B: Intermediate 107 was prepared using intermediate 106 (400.00 mg) and NBS (168.35 mg, 945.91 umol, 1.03 eq) as raw material through a similar procedure to that described in preparing intermediate 19.
(250) Step C: I-54 was prepared using intermediate 107 (400.00 mg, 1 eq) and trimethylboroxine (300.13 mg, 2.39 mmol, 333.48 uL, 3.00 eq) as raw material through a similar procedure to that described in preparing I-19. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.78 (s, 1H), 7.48 (d, J=7.6 Hz, 1H), 7.37-7.27 (m, 4H), 7.13-6.89 (m, 8H), 6.37 (d, J=16 Hz, 1H), 2.42 (t, J=7.6 Hz, 2H), 2.23 (s, 3H), 2.19 (s, 3H), 0.86 (t, J=7.6 Hz, 3H); MS (ESI, M+1): 422.2.
Embodiment 62
(251) ##STR00151##
(252) I-55 was prepared using intermediate 51 (300.00 mg, 0.656 mmol, 1.00 eq) and 2-chloro-5-bromo-6-cyanopyridine (214.21 mg, 985.10 umol, 1.5 eq) as raw material through a similar procedure to that described in preparing I-28.
(253) The crude product was purified by preparative high performance liquid chromatography (formic acid system) to give the product I-55. .sup.1H NMR (400 MHz, DMSO-d.sub.6): 11.46 (brs, 1H), 8.25 (d, J=8.4 Hz, 1H), 7.94 (d, J=8.4 Hz, 1H), 7.61-7.34 (m, 5H), 7.26-7.13 (m, 2H), 6.96 (d, J=7.6 Hz, 1H), 6.45 (brs, 1H), 2.58 (q, J=7.2 Hz, 2H), 0.98 (t, J=7.2 Hz, 3 H). MS (ESI, M+1): 488.0.
Embodiment 63
(254) ##STR00152##
(255) I-56 was prepared using intermediate 12 (1000 mg, 1.85 mmol, 1.00 eq) and 5-methyl-3-bromothiophene (588.21 mg, 3.32 mumol, 1.8 eq) as raw material through a similar procedure to that described in preparing I-24.
(256) The crude product was purified by preparative high performance liquid chromatography (formic acid system) to give the product I-56. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 11.43 (s, 1H), 7.53-7.43 (m, 4H), 7.36 (d, J=8.0 Hz, 1H), 7.22-7.09 (m, 2H), 7.06-6.98 (m, 3H), 6.53 (s, 1H), 6.44 (d, J=16 Hz, 1H), 2.39 (q, J=7.2 Hz, 2H), 2.34 (s, 3H), 0.96 (t, J=7.2 Hz, 3H); MS (ESI, M+1): 448.0.
Embodiment 64
(257) ##STR00153##
(258) Step A: Methyl acrylate (25.32 g, 294.10 mmol, 26.38 mL, 5.00 eq), triethylamine (11.90 g, 117.64 mmol, 16.30 mL, 2.00 eq) and 1,1-bis(diphenylphosphino)ferrocene palladium chloride (2.15 g, 2.94 mmol, 0.05 eq) were added to a solution of 2,6-difluoro-4-iodoaniline (15.00 g, 58.82 mmol, 1.00 eq) in 100 mL N,N-dimethylformamide. The reaction solution was stirred at 80 C. under nitrogen atmosphere for 12 hours. Then 400 mL ethyl acetate was added, and the mixture was filtered through celite. The filtrate was washed three times with 400 mL water. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give intermediate 108.
(259) Step B: Iodine (19.64 g, 77.40 mmol, 15.59 mL, 3.00 eq) was added to a solution of 108 (5.50 g, 25.80 mmol, 1.00 eq) in 50 mL acetonitrile, followed by addition of tert-butyl nitrite (3.99 g, 38.70 mmol, 4.59 mL, 1.50 eq) at 0 C. under nitrogen atmosphere. The reaction solution was stirred at 15 C. under nitrogen atmosphere for 12 hours. Then 150 mL ethyl acetate was added, and the mixture was filtered through celite. The filtrate was washed twice with 200 mL saturated aqueous sodium sulfite. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give intermediate 109.
(260) Step C: Aqueous potassium hydroxide solution (4M, 3.33 mL, 5.00 eq) and bis(triphenylphosphine)palladium dichloride (93.44 mg, 133.00 umol, 0.05 eq) were added to a solution of 30 (1.40 g, 2.66 mmol, 1.00 eq) in 30 mL dimethyltetrahydrofuran. The reaction solution was stirred at 70 C. under nitrogen atmosphere for 12 hours, then filtered through celite. The filtrate was neutralized to pH 5 with hydrochloric acid (1M). 40 mL water was added, then extracted twice with 30 mL ethyl acetate. The organic phase was combined, washed twice with 30 mL water, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative HPLC (formic acid system) to give the product I-57. MS(ESI, M+1): 482.1 .sup.1H NMR EW3644-119-P1E (400 MHz, DMSO-d.sub.6): 12.52 (brs, 1H), 10.81 (s, 1H), 7.54 (d, J=8.0 Hz, 1H), 7.45 (d, J=16.4 Hz, 1H), 7.36-7.32 (m, 4H), 7.23 (ddd, J=2.0 Hz, J=6.4 Hz, J=8.8 Hz, 1 H), 7.13 (dt, J=2.8 Hz, J=8.4 Hz, 1 H), 7.07 (dt, J=1.2 Hz, J=7.2 Hz, 1 H), 6.99 (dt, J=0.8 Hz, J=8.0 Hz, 1 H), 6.59 (d, J=16.0 Hz, 1 H), 6.55 (d, J=1.2 Hz, 1 H), 2.88-2.81 (m, 2 H), 1.05 (t, J=7.6 Hz, 3 H).
Embodiment 65
(261) ##STR00154##
(262) II-12 was prepared using intermediate 10 (500.00 mg, 2.92 mmol, 1.00 eq) and intermediate (920 mg, 2.56 mmol, 0.9 eq) and o-cyano-p-chlorobromobenzene (922 mg, 4.26 mmol) as raw material through a similar procedure to that described in preparing II-8.
(263) The crude product was purified by preparative high performance liquid chromatography (formic acid system) to give the product I-56. .sup.1H NMR (400 MHz, DMSO-d.sub.6) =11.37 (s, 1H), 7.94 (d, J=2.4 Hz, 1H), 7.78-7.75 (m, 1H), 7.53-7.51 (m, 2H), 7.39-7.37 (m, 1H), 7.20-7.08 (m, 2H), 6.82 (d, J=8.8 Hz, 2H), 6.66 (d, J=8.8 Hz, 2H), 3.78 (t, J=5.2 Hz, 2H), 3.35-3.31 (m, 2H), 2.29 (t, J=5.2 Hz, 2H), 2.72-2.70 (m, 2H), 2.56-2.55 (m, 5H), 0.90 (t, J=7.6 Hz, 3H); MS (ESI, M+1): 555.2.
(264) Effect Embodiment 1
(265) (1) MCF-7 Cell Proliferation Inhibition Assay
(266) Experimental Reagent:
(267) RPMI 1640 medium, fetal bovine serum, Promega CellTiter-Glo reagent. The cell line MCF-7 was purchased from The European Collection of Authenticated Cell Cultures (ECACC). Envision Multilabel Reader (PerkinElmer).
(268) Experimental Method:
(269) MCF-7 cells were seeded in black 384-well plates at 600 cells per well in 30 mL cell suspension. The cell plate was placed in a carbon dioxide incubator overnight.
(270) The test compound was diluted 5-fold to the 10th concentration with Epmotion, that was diluting from 2.5 mM to 1.28 nM, each concentration set two replicated wells. 198 L medium was added to the intermediate plate, the compound with gradient concentrations was transferred to the intermediate plate at 2 L per well according to the corresponding position, then mixing and transferring to the cell plate at 20 L per well. The cell plate was incubated in a carbon dioxide incubator for 6 days.
(271) Promega CellTiter-Glo reagent was added to the cell plate at 25 L per well, cells were incubated for 10 minutes at room temperature to stabilize the luminescence signal, which was measured using PerkinElmer Envision Multilabel Reader.
(272) Data Analysis:
(273) The original data was converted to the inhibition rate using the equation (MaxRatio)/(MaxMin)*100%, and the value of IC.sub.50 could be obtained by curve fitting with four parameters. (Model 205 in XLFIT5, iDBS)
(274) (2) ER Degradation Assay in MCF-7 Cells
(275) Experimental Reagent:
(276) RPMI 1640 medium, fetal bovine serum, PBS, 16% paraformaldehyde, Triton, blocking solution, estrogen receptor antibody, near-infrared goat anti-rabbit secondary antibody, DRAQ5 dye. The cell line MCF-7 was purchased from The European Collection of Authenticated Cell Cultures (ECACC). Odyssey infrared fluorescence scanning imaging system.
(277) Experimental Method:
(278) MCF-7 cells were seeded in black 384-well plates at 3200 cells per well in 30 mL cell suspension. The cell plate was placed in a carbon dioxide incubator for 4 days.
(279) The test compound was diluted 5-fold to the 10th concentration with Epmotion, fulvestrant was diluted from 0.25 mM to 0.128 nM, other compounds were diluted from 2.5 mM to 1.28 nM, each concentration set two replicated wells. 198 L medium was added to the intermediate plate, the compound with gradient concentrations was transferred to the intermediate plate at 2 L per well according to the corresponding position, then mixing and transferring to the cell plate at 20 L per well. The cell plate was incubated in a carbon dioxide incubator for 20 hours.
(280) After 8% paraformaldehyde was added to the cell plate at 50 L per well, the cell plate was incubated at room temperature for 30 minutes, then washed twice with PBS. After pat dry, 50 L PBS containing 0.1% Triton was added, and the cell plate was incubated at room temperature for 15 minutes, then washed with PBS for five times. After 50 L blocking solution was added, the cell plate was incubated at room temperature for 1 hour. After pat dry, 50 uL blocking solution containing 0.1% estrogen receptor antibody was added, and the cell plate was incubated at 4 C. overnight. The next day, after the primary antibody was brushed down, the cell plate was washed with PBS for five times, then a blocking solution containing 0.1% near-infrared goat anti-rabbit secondary antibody and 0.05% DRAQ5 dye was added. The cell plate was incubated at room temperature for 1 hour, washed with PBS for five times. After pat dry, the cell plate was measured using Odyssey infrared fluorescence scanning imaging system.
(281) Data Analysis:
(282) The original data was converted to the inhibition rate using the equation (MaxRatio)/(MaxMin)*100%, and the value of IC.sub.50 could be obtained by curve fitting with four parameters. (Model 205 in XLFIT5, iDBS)
(283) TABLE-US-00001 TABLE 1 screening test results in vitro anti-proliferation ER degradation assay in MCF-7 cells in MCF-7 cells Compound (nM) IC.sub.50 (nM) I-1 38.5 20.8 I-2 282 68 I-3 206 93 I-4 325 131 I-5 8.04 5.87 I-6 321 64 I-7 9.52 7.88 I-8 5.59 4.49 I-9 167 93.67 I-10 13 9.95 I-11 8.53 2.44 I-12 13 5.59 I-13 27 5.83 I-14 4.11 2 I-15 7.56 2.25 I-17 8.29 6.23 I-18 22 38 I-19 4.78 1.26 I-21 14 18 I-22 0.86 0.81 I-23 4.84 2.6 I-24 21.5 7.56 I-27 8.36 13 I-28 69 64 I-29 30 42 I-30 48 20 I-31 3.66 2.66 I-32 61 30 I-33 2.86 2.35 I-34 49 31 I-35 20 5.13 I-36 6.84 1.78 I-37 11 9 I-40 2.31 2.84 I-41 3.91 2.54 I-42 0.77 1.57 I-43 1.56 0.70 I-44 5.84 12 I-45 103 178 I-46 393 220 I-47 4.27 4.13 I-48 4.88 5.77 I-49 42 29 I-50 33 9 I-51 9.24 9 I-52 40 49 I-53 4.35 2.02 I-54 23 4.89 I-55 2.42 2.82 I-56 35 18 I-57 32 18.5 II-1 4.57 5.36 II-2 7.75 34.34 II-3 6.77 12.5 II-4 6.86 26 II-5 7.23 5.25 II-6 2.46 5.28 II-7 2.61 3.41 II-8 5 26 II-9 51 23 II-10 3.92 9.88 II-11 7.43 3.93 II-12 1.56 5.51
(284) Conclusion:
(285) The compound of the present invention exhibits excellent in vitro activity.
(286) Effect Embodiment 2: Evaluation of In Vitro DMPK Properties
(287) (1) The results of in vitro DMPK properties are shown in table 2:
(288) TABLE-US-00002 TABLE 2 results of in vitro DMPK properties Compound ARN-810 I-1 plasma protein binding rate NA (too high to 97.2%/98.8% (Human/CD-1 mouse) be detected) inhibition of CYP enzymes (IC.sub.50, 13.7/0.664/0.884/ 25.5/10.1/40.4/ M) 1A2/2C9/2C19/2D6/3A4 23.6/24.6 43.4/25.1
(289) Conclusion:
(290) Compound I-1 of the present invention is superior to oral selective estrogen receptor down-regulator ARN-810 which is in clinical phase II in terms of plasma protein binding rate (PPB) and drug-drug interaction (DDI).
(291) (2) The results of in vivo PK properties are shown in table 3:
(292) TABLE-US-00003 TABLE 3 results of in vivo PK properties Compound ARN-810 I-1 Cassette PK i.V. P.O i.V. P.O (1 mpk) (2.5 mpk) (1 mpk) (2.5 mpk) C.sub.0 (For i.v.) [C.sub.max 1639 924/0.417 3479 2147/3.33 (For P.O)/T.sub.max] (nM) T.sub.1/2 (hr) 1.35 3.49 2.81 3.18 Vdss (L/Kg) 1.93 N.D. 0.992 N.D. Cl (mL/min/Kg) 18.9 N.D. 3.61 N.D. AUC.sub.0-last (nm/hr) 1987 3277 10378 18327 F % 66 70.6
(293) Conclusion:
(294) Compound I-1 of the present invention is superior to oral selective estrogen receptor down-regulator ARN-810 in clinical phase II in terms of oral peak plasma concentration Cmax, half-life, clearance rate (CL), AUC and oral bioavailability.
(295) Effect Embodiment 3: Evaluation of In Vivo Pharmacodynamics
(296) The purpose of this experiment was to evaluate antitumor activity of oral selective estrogen receptor down-regulator ARN-810 in clinical phase II and compound I-1 on MCF-7 breast cancer cell xenograft model in BALB/c nude mice (Beijing Vital River Laboratory Animal Technology Co., Ltd., 10 animals per experimental group). In this experiment, mice inoculated with MCF-7 breast cancer cells were orally administered with ARN-810 at a dose of 10/30 mg/kg, with compound I-1 at a dose of 30 mg/kg, and the inhibitory activity of the test drug on the tumor growth was evaluated.
(297) Mice were inoculated subcutaneously with 0.36 mg estrogen sustained release tablet (60 days release) on the left shoulder three days before inoculation. When cells were in the logarithmic growth phase, cells were harvested, counted and adjusted to 1010.sup.7 cells/mL, then an equal volume of Matrigel was added, mixing for inoculation. 0.2 mL MCF-7 tumor cell suspension (1010.sup.6) was inoculated subcutaneously into the right shoulder of each mouse. On the 7th day after tumor cell inoculation, mice were grouped and administrated once a day, with an average tumor volume of 169 mm.sup.3 and a body weight of 23.33 g. Tumor volume and body weight were measured twice a week after grouping, and the tumor growth rate (T/C) and tumor growth inhibition rate (TGI) were calculated based on the last measuring data on the 17th day after grouping. The results are as follows:
(298) TABLE-US-00004 TABLE 4 evaluation of antitumor activity tumor volumn (mm.sup.3)a T/C TGI Group test compound 17th day after grouping (%) (%) P 1 ARN-810 290 38 43.4 75.7 <0.001 2 I-1 201 27 30.1 93.6 <0.001 aMean SEM.
(299) Conclusion:
(300) The antitumor activity of oral administration of I-1 at a dose of 30 mg/kg is superior to that of oral administration of oral selective estrogen receptor down-regulator ARN-810 in clinical phase II at a dose of 10/30 mg/kg.
(301) Effect Embodiment 4: Evaluation of Drug Distribution in Tissues
(302) On the 20th day after grouping, the sample was collected from plasma, tumor, breast and brain of the mice at four time points (2-3 mice each time point) which was respectively 0.5 hour, 1.5 hours, 6 hours and 24 hours after administration. The blood (0.5 mL) was collected using a 1.5 mL EDTA-K2 anticoagulation tube, immediately centrifuged at 4000 rpm, 4 C. for 10 minutes to prepare plasma, the plasma sample was stored at 80 C. for determination of drug concentration. The tumor was collected using a 2 mL cryotube, and the brain was collected using a 5 mL cryotube, the samples were immediately put into liquid nitrogen for quike freeze. All samples were stored at 80 C. for determination of drug concentration after sampling. The result was converted based on corresponding AUC to compare the drug distribution.
(303) The result was shown in table 5.
(304) TABLE-US-00005 TABLE 5 evaluation of drug distribution in tissues AUC0-last (nmol .Math. h/kg) molecular dose Compound weight (mol/kg) plasma tumor brain ARN-810 446.91 67.3 33545 11368 3490 I-1 445.91 67.3 118937 55487 43167
(305) Conclusion:
(306) The distribution of compound I-1 in tumor tissue is much better than that of ARN-810, which is consistent with its better antitumor activity;
(307) The distribution of compound I-1 in brain tissue is much better than that of ARN-810, indicating that it will have excellent activity to treat metastatic ER-positive breast cancer in brain.