A pyrimidine pyridine CDK inhibitor and its preparation method and application

Abstract

The present invention discloses a pyrimidine pyridine CDK inhibitor and a preparation method and application thereof, wherein the compound is shown in formula (S). The compound of the present invention can inhibit the continuous proliferation of cancer cells and can be used to prepare drugs for treating cancer or tumor-related diseases.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry, and in particular to a pyrimidine-pyridine CDK inhibitor and a preparation method and application thereof. Background Art

[0002] Cyclin-dependent kinases (CDKs) control cell proliferation by regulating the progression of the cell cycle. When CDKs are abnormally activated, cells will continue to proliferate uncontrollably, which is one of the important signs of cancer. CDK6 inhibitors inhibit the malignant proliferation of cancer cells by regulating the transition of the cell cycle from the G1 phase to the S phase, and play an important role in cancer treatment. CDK2 also plays an important role in the transition of the cell cycle from the G1 phase to the S phase. Therefore, inhibiting CDK2 or CDK6 can produce significant cell proliferation inhibitory activity. Summary of the invention

[0003] Purpose of the invention: The present invention provides a pyrimidine pyridine CDK inhibitor, which inhibits CDK2 or CDK6. The present invention also provides a preparation method and application of the compound.

[0004] Technical solution: The compound of the present invention as shown in formula (S) or a pharmaceutically acceptable salt thereof:

[0005]

[0006] in,

[0007] R1 is selected from OR6 or NR7R8, wherein R6 is selected from C1-C3 alkyl, and R7 and R8 are selected from hydrogen or C1-C3 alkyl;

[0008] R2 is selected from hydrogen, halogen, -COOH or C1-C3 alkyl;

[0009] R3 is selected from hydrogen or C1-C3 alkyl;

[0010] R4 is selected from hydrogen, halogen or -OC1-C3 alkoxy;

[0011] R5 is selected from -CH2CH2OCH3, -C1-C5 alkyl or C3-C4 cycloalkyl.

[0012] Preferably:

[0013] The R1 is selected from OR6 or NR7R8, wherein R6 is selected from methyl, ethyl, isopropyl, R7 is selected from hydrogen or methyl, and R8 is selected from methyl or isopropyl;

[0014] Said R2 is selected from hydrogen, fluorine, -COOH, methyl or ethyl;

[0015] Said R3 is selected from hydrogen, methyl or ethyl;

[0016] Said R4 is selected from hydrogen, halogen or -OC1-C3 alkoxy;

[0017] The R5 is selected from -CH2CH2OCH3, -C1-C5 alkyl or C3-C4 cycloalkyl.

[0018] Preferably:

[0019] The R1 is selected from OR6 or NR7R8, wherein R6 is selected from methyl, ethyl, isopropyl, R7 is selected from hydrogen or methyl, and R8 is selected from methyl or isopropyl;

[0020] The R2 is selected from hydrogen, fluorine, -COOH, and methyl;

[0021] Said R3 is selected from hydrogen and methyl;

[0022] Said R4 is selected from hydrogen, fluorine or -OC1-C3 alkoxy;

[0023] The R5 is selected from -CH2CH2OCH3, -C1-C5 alkyl or C3-C4 cycloalkyl.

[0024] Preferably:

[0025] The R1 is selected from OR6 or NR7R8, wherein R6 is selected from methyl, ethyl, isopropyl, R7 is selected from hydrogen or methyl, and R8 is selected from methyl or isopropyl;

[0026] Said R2 is selected from hydrogen, fluorine, -COOH or methyl;

[0027] Said R3 is selected from hydrogen or methyl;

[0028] Said R4 is selected from hydrogen, fluorine or methoxy;

[0029] The R5 is selected from -CH2CH2OCH3, -C1-C4 alkyl or C3-C4 cycloalkyl.

[0030] Preferably:

[0031] The R1 is selected from OR6 or NR7R8, wherein R6 is selected from methyl, ethyl, isopropyl, R7 is selected from hydrogen or methyl, and R8 is selected from methyl or isopropyl;

[0032] Said R2 is selected from hydrogen, fluorine, -COOH or methyl;

[0033] Said R3 is selected from hydrogen or methyl;

[0034] Said R4 is selected from hydrogen, fluorine or methoxy;

[0035] The R5 is selected from -CH2CH2OCH3, -C1-C4 alkyl or cyclopropyl.

[0036] Preferably:

[0037] The R1 is selected from OR6 or NR7R8, wherein R6 is selected from methyl, ethyl, isopropyl, R7 is selected from hydrogen or methyl, and R8 is selected from methyl or isopropyl;

[0038] Said R2 is selected from hydrogen, fluorine, -COOH or methyl;

[0039] Said R3 is selected from hydrogen or methyl;

[0040] Said R4 is selected from hydrogen, fluorine or methoxy;

[0041] The R5 is selected from -CH2CH2OCH3, methyl, ethyl, n-propyl, isopropyl or cyclopropyl.

[0042] In some specific embodiments of the present invention, the present invention also provides a compound selected from S-1 to S-17 or a pharmaceutically acceptable salt thereof, selected from the following compounds:

[0043]

[0044]

[0045]

[0046] The above-mentioned pharmaceutically acceptable salts are acid addition salts of the compound of general formula (S), wherein the acid used for salt formation includes inorganic acids and organic acids, the inorganic acids include: hydrochloric acid, sulfuric acid, phosphoric acid and methanesulfonic acid, and the organic acids include acetic acid, trichloroacetic acid, propionic acid, butyric acid, maleic acid, p-toluenesulfonic acid, malic acid, malonic acid, cinnamic acid, citric acid, fumaric acid, camphoric acid, digluconic acid, aspartic acid and tartaric acid.

[0047] Preferably, the pharmaceutically acceptable salt described in the present invention is hydrochloride.

[0048] The preparation method of the compound of general formula (S) of the present invention,

[0049] Compound A and compound B are reacted with each other under the action of a palladium catalyst to prepare compound S;

[0050]

[0051] or compound C reacts with compound D under the action of a base to prepare compound S;

[0052]

[0053] R1 is selected from OR6 or NR7R8, wherein R6 is selected from C1-C3 alkyl, and R7 and R8 are selected from hydrogen or C1-C3 alkyl;

[0054] R2 is selected from hydrogen, halogen, -COOH or C1-C3 alkyl;

[0055] R3 is selected from hydrogen or C1-C3 alkyl;

[0056] R4 is selected from hydrogen, halogen or -OC1-C3 alkoxy;

[0057] R5 is selected from -CH2CH2OCH3, -C1-C5 alkyl or C3-C4 cycloalkyl.

[0058] The present invention also provides a pharmaceutical composition of a compound as described in formula (S) or a pharmaceutically acceptable salt, ester, stereoisomer, solvent compound or prodrug thereof, and a pharmaceutically acceptable carrier.

[0059] A pharmaceutically acceptable carrier refers to an excipient or diluent that does not cause significant irritation to an organism and does not interfere with the biological activity and properties of the administered compound.

[0060] The present invention also provides a use of a compound of formula (S) or a pharmaceutically acceptable salt, ester, stereoisomer, solvent compound or prodrug thereof or the composition of the present invention in preparing a drug.

[0061] The present invention also provides a use of a compound of formula (S) or a pharmaceutically acceptable salt, ester, stereoisomer, solvent compound or prodrug thereof or the composition of the present invention in the preparation of a CDK2 or CDK6 inhibitor; the CDK2 or CDK6 kinase inhibitor is used to treat cancer or tumor-related diseases.

[0062] Use of the compound of the present invention or its pharmaceutically acceptable salt, ester, stereoisomer, solvate or prodrug or the composition of the present invention in the preparation of a drug for treating cancer or tumor-related diseases. The cancer or tumor is a cancer or tumor related to CDK2 or CDK6. Cancer or tumor-related diseases include but are not limited to leukemia, breast cancer, prostate cancer, lung cancer, multiple myeloma, liver cancer, gastric cancer, bone cancer, brain cancer, head and neck cancer, intestinal cancer, pancreatic cancer, bladder cancer, testicular cancer, ovarian cancer, endometrial cancer, etc.

[0063] The compound of the general formula (S) or a pharmaceutically acceptable salt thereof described in the present invention has CDK2 or CDK6 target inhibitory activity and has a therapeutic effect on malignant cell proliferation tumors.

[0064] Beneficial effects: The compound represented by the general formula (S) of the present invention can inhibit the malignant proliferation of cancer cells, has good therapeutic effect, low toxicity, good drug metabolism characteristics, and is not prone to drug resistance. It can be used to prepare drugs for treating cancer or tumor-related diseases. DETAILED DESCRIPTION

[0065] The following examples are provided to facilitate a better understanding of the present invention, but are not intended to limit the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples are purchased from conventional biochemical reagent stores unless otherwise specified. The present application is described in detail below in conjunction with specific embodiments.

[0066] 1. Synthesis of intermediate reactants

[0067] The specific preparation method of the independently developed intermediate reactant (M) is as follows:

[0068] (1) Synthesis of 5-bromo-3-fluoro-2-isopropoxypyridine (M1)

[0069]

[0070] 3-Fluoro-5-bromo-2-pyridone (3.84 g, 20 mmol) and silver carbonate (16.55 g, 60 mmol) were dissolved in toluene (200 mL), and then 2-iodopropane (10.2 g, 60 mmol) was added, heated to 150°C, reacted for 15 min, and purified by rapid silica gel column to obtain compound M1 (3.74 g, yield 80%). 1 H NMR (400MHz, Chloroform-d) δ7.27-7.26(m,1H),7.16(dd,J=8.4,2.5Hz,1H),5.31-5.24(m,1H),1.38(d,J=6.8Hz,6H).

[0071] (2) Synthesis of 2-chloro-4-(5-fluoro-6-isopropoxypyridin-3-yl)pyrimidine (M2)

[0072]

[0073] Compound M1 (3.74 g, 16.0 mmol), bis(pyrimidine) (4.47 g, 17.6 mmol), dissolved in 1,4-dioxane (100 mL), then added Pd(dppf)Cl2 (584.8 mg, 0.8 mmol), potassium acetate (4.71 g, 48 mmol), replaced argon three times, heated to 100 ° C, reacted for 12 h. After the reaction was completed, water was added to quench, ethyl acetate was used, and the compound M1-2 was obtained by evaporation under reduced pressure and concentrated, without purification, and used directly in subsequent reactions. 2,4-dichloropyrimidine (2.86 g, 19.2 mmol), Pd(PPh3)2Cl2 (561.6 mg, 0.8 mmol), sodium carbonate (4.24 g, 40 mmol), water (20 ml) and ethylene glycol dimethyl ether (25 mL) were added to a three-necked flask, and argon was replaced three times. Compound M1-2 dissolved in ethylene glycol dimethyl ether (120 mL) was slowly added into a three-necked flask, heated to 80° C., reacted for 12 h, and purified by rapid silica gel column to obtain compound M2 (2.91 g, yield 68%). 1 H NMR(400MHz,Chloroform-d)δ8.63(d,J=5.3Hz,1H),8.61(d,J=2.1Hz,1H),8.11(dd, J=10.8,2.1Hz,1H),7.59(d,J=5.3Hz,1H),5.51-5.45(m,1H),1.44(d,J=6.2Hz,6H).

[0074] (3) Synthesis of 2-chloro-4-(5-fluoro-6-isopropoxypyridin-3-yl)-5-methoxypyrimidine (M3)

[0075]

[0076] Referring to the synthesis method of compound M2, the yield was 56%. 1 H NMR (400MHz, Chloroform-d) δ8.75(d,J=2.1Hz,1H),8.20(s,1H),8.09(dd,J=11.6,2.1Hz,1H),5.41-5.35(m,1H),3.96(s,3H),1.34(d,J=6.3Hz,6H).

[0077] (4) Synthesis of 2-chloro-5-fluoro-4-(5-fluoro-6-isopropoxypyridin-3-yl)pyrimidine (M4)

[0078]

[0079] Referring to the synthesis method of compound M2, the yield was 58%. 1H NMR (400MHz, Chloroform-d) δ8.77-8.76(m,1H),8.52(d,J=3.0Hz,1H),8.17(dd,J=11.1,2.1Hz,1H),5.54-5.47(m,1H),1.45(d,J=6.3Hz,6H).

[0080] (5) Synthesis of tert-butyl 4-((5-fluoro-4-(5-fluoro-6-isopropoxypyridin-3-yl)pyrimidin-2-yl)amino)piperidine-1-carboxylate (M5)

[0081]

[0082] Compound M4 (571 mg, 2 mmol) and 1-Boc-4-aminopiperidine (500 mg, 2.5 mmol) were dissolved in dioxane (6 mL), and then Pd2(dba)3 (92 mg, 0.1 mmol), Xantphos (116 mg, 0.2 mmol), and cesium carbonate (1.3 g, 4 mmol) were added, and argon was replaced three times, heated to 105°C, and reacted for 12 h. Compound M5 (432 mg, 48% yield) was obtained by cooling, filtering, concentrating, and column chromatography as a white solid. MS (M+H) + :found,450.6.

[0083] (6) Synthesis of 5-fluoro-4-(5-fluoro-6-isopropoxypyridin-3-yl)-N-(piperidin-4-yl)pyrimidin-2-amine (M6)

[0084]

[0085] Compound M5 (899 mg, 2 mmol) was dissolved in dichloromethane (10 mL), 2N hydrogen chloride-ethyl acetate solution (5 mL) was added, and the mixture was reacted at room temperature for 4 h. The pH value of the reaction solution was adjusted to neutral, and compound M6 (664 mg, 95% yield) was obtained by column chromatography as a white solid. 1 H NMR (400MHz, DMSO-d6) δ8.64(s,1H),8.48(d,J=3.6Hz,1H),8.18(d,J=11.7Hz,1H),7.56(d,J=7.3Hz,1H),5.45-5.39(m,1 H),4.08-3.96(m,1H),3.30-3.26(m,2H),3.05-2.96(m,2H),2.07-2.03(m,2H),1.76-1.67(m,2H),1.37(d,J=6.2Hz,6H).

[0086] (7) Synthesis of 5-bromo-2-isopropoxypyridine (M7)

[0087]

[0088] 3-Fluoro-5-bromo-2-pyridone was replaced by 5-bromopyridin-2(1H)-one, and the yield was 55% according to the synthesis method of compound M1. 1 H NMR (400MHz, Chloroform-d) δ7.47(d,J=2.7Hz,1H),7.32(dd,J=9.6,2.7Hz,1H),6.48(d,J=9.6Hz,1H),5.27-5.16(m,1H),1.37(d,J=6.9Hz,6H). 13 C NMR (75MHz, CDCl3) δ162.25,147.44,140.99,113.24,111.04,68.55,21.94.

[0089] (8) Synthesis of 2-chloro-5-fluoro-4-(6-isopropoxypyridin-3-yl)pyrimidine (M8)

[0090]

[0091] Referring to the synthesis method of compound M2, the yield was 65%. 1 H NMR(400MHz,Chloroform-d)δ8.98-8.97(m,1H),8.49(d,J=3.1Hz,1H),8.39-8. 36(m,1H),6.81(dd,J=8.9,0.7Hz,1H),5.45-5.39(m,1H),1.39(d,J=6.2Hz,6H).

[0092] (9) Synthesis of 5-bromo-2-ethoxypyridine (M9)

[0093]

[0094] The yield was 70% by replacing isopropyl iodide with ethyl iodide and referring to the synthesis method of compound M1. 1 H NMR (400MHz, Chloroform-d) δ7.41(d,J=2.7Hz,1H),7.34(dd,J=9.6,2.7Hz,1H),6.49(d,J=9.6Hz,1H),3.96(q,J=7.2Hz,2H),1.36(t,J=7.2Hz,3H).

[0095] (10) Synthesis of 2-chloro-4-(6-ethoxypyridin-3-yl)-5-fluoropyrimidine (M10)

[0096]

[0097] Referring to the synthesis method of compound M2, the yield was 70%. 1 H NMR(400MHz,Chloroform-d)δ8.99-8.98(m,1H),8.50(d,J=3.1Hz,1H),8.41-8.38 (m,1H),6.87(dd,J=8.9,0.8Hz,1H),4.46(q,J=7.1Hz,2H),1.44(t,J=7.1Hz,3H).

[0098] (11) Synthesis of 5-bromo-2-isopropoxy-3-methylpyridine (M11)

[0099]

[0100] The yield was 54% by replacing 3-fluoro-5-bromo-2-pyridone with 5-bromo-2-hydroxy-3-methylpyridine and referring to the synthesis method of compound M1. 1 H NMR (400MHz, Chloroform-d) δ7.30 (d, J = 2.7Hz, 1H), 7.23-7.21 (m, 1H), 5.31-5.21 (m, 1H), 2.15 (s, 3H), 1.35 (d, J = 6.8Hz, 6H).

[0101] (12) Synthesis of 2-chloro-5-fluoro-4-(6-isopropoxy-5-methylpyridin-3-yl)pyrimidine (M12)

[0102]

[0103] Referring to the synthesis method of compound M2, the yield was 63%. 1 H NMR (400MHz, Chloroform-d) δ8.80 (s, 1H), 8.46 (d, J = 3.2Hz, 1H), 8.18-8.17 (m, 1H), 5.46-5.40 (m, 1H), 2.24 (s, 3H), 1.39 (d, J = 6.2Hz, 6H). 13 C NMR (126MHz, CDCl3) δ164.24,155.75,155.52,153.64,153.05,152.97,147.95,147 .74,146.67,146.57,138.04,138.01,121.82,120.75,120.70,69.14,22.12,15.94.

[0104] (13) Synthesis of 3-bromo-6-isopropoxy-2-methylpyridine (M13)

[0105]

[0106] 3-Fluoro-5-bromo-2-pyridone was replaced with 5-bromo-6-methyl-2(1H)-pyridone, and the yield was 63% according to the synthesis method of compound M1. 1 H NMR (400MHz, Chloroform-d) δ7.58(d,J=8.6Hz,1H),6.39(d,J=8.6Hz,1H),5.29-5.19(m,1H),2.52(s,3H),1.32(d,J=6.2Hz,6H).

[0107] (14) Synthesis of 2-chloro-5-fluoro-4-(6-isopropoxy-2-methylpyridin-3-yl)pyrimidine (M14)

[0108]

[0109] Compound M13 (2.30 g, 10.0 mmol), bis(pyrimidine) (2.79 g, 11 mmol), dissolved in 1,4-dioxane (80 mL), then added Pd(dppf)Cl2 (365.5 mg, 0.5 mmol), potassium acetate (2.94 g, 30 mmol), replaced argon three times, heated to 100 ° C, reacted for 12 h. After the reaction was completed, water was added to quench, ethyl acetate was used, and the compound M13-2 was obtained by evaporation under reduced pressure and concentrated, without purification, and used directly in subsequent reactions. 2,4-dichloro-5-fluoropyrimidine (2.00 g, 12 mmol), Pd(PPh3)2Cl2 (351 mg, 0.5 mmol), sodium carbonate (2.65 g, 25 mmol), water (15 ml) and ethylene glycol dimethyl ether (20 mL) were added to a three-necked flask, and argon was replaced three times. Compound M13-2 dissolved in ethylene glycol dimethyl ether (80 mL) was slowly added into a three-necked flask, heated to 80° C., reacted for 12 h, and purified by rapid silica gel column to obtain compound M14 (1.83 g, yield 65%). 1 H NMR(400MHz,Chloroform-d)δ8.52(d,J=1.7Hz,1H),7.67(dd,J=8.6,1.3Hz,1H),6. 63(d,J=8.6Hz,1H),5.43-5.37(m,1H),2.47(d,J=2.3Hz,3H),1.37(d,J=6.2Hz,6H). 13C NMR(101MHz,DMSO)δ163.59,156.53,156.14,156.00,155.54,154.43,154.40,153.94, 149.61,149.35,141.43,141.40,119.67,119.64,108.93,68.49,23.11,23.07,22.21.

[0110] (15) Synthesis of 5-bromo-2-isopropoxynicotinate methyl ester (M15)

[0111]

[0112] The yield was 56% by replacing 3-fluoro-5-bromo-2-pyridone with 5-bromo-2-hydroxynicotinate methyl ester and referring to the synthesis method of compound M1. 1 H NMR (400MHz, Chloroform-d) δ8.30(d,J=2.6Hz,1H),8.20(d,J=2.6Hz,1H),5.39-5.33(m,1H),3.89(s,3H),1.38(d,J=6.3Hz,6H). 13 C NMR (101MHz, CDCl3) δ163.79,160.41,150.97,142.81,115.31,109.86,69.61,51.95,21.66.

[0113] (16) Synthesis of 5-(2-chloro-5-fluoropyrimidin-4-yl)-2-isopropoxynicotinic acid methyl ester (M16)

[0114]

[0115] The synthetic method of reference compound M14 had a yield of 61%. 1 H NMR (400MHz, Chloroform-d) δ9.09-9.08(m,1H),8.91(d,J=2.5Hz,1H),8.53(d,J=3.0Hz,1H),5.59-5.50(m,1H),3.94(s,3H),1.44(d,J=6.2Hz,6H).

[0116] (17) Synthesis of 5-bromo-3-fluoro-2-methoxypyridine (M17)

[0117]

[0118] The yield of M1 was 51% when isopropyl iodide was replaced by methyl iodide. 1H NMR (400MHz, Chloroform-d) δ7.99 (d, J=2.1Hz, 1H), 7.47 (dd, J=9.3, 2.1Hz, 1H), 4.00 (s, 3H). 13 C NMR (101MHz, CDCl3) δ152.54,152.43,148.27,145.63,141.95,141.89,126.17,125.99,110.00,54.02.

[0119] (18) Synthesis of 2-chloro-5-fluoro-4-(5-fluoro-6-methoxypyridin-3-yl)pyrimidine (M18)

[0120]

[0121] The synthetic method of reference compound M14 had a yield of 66%. 1 H NMR (400MHz, DMSO-d6) δ9.00(d,J=3.1Hz,1H),8.67(dd,J=2.0,1.2Hz,1H),8.21(dd,J=11.3,2.0Hz,1H),4.06(s,3H).

[0122] (19) Synthesis of 5-bromo-2-ethoxy-3-fluoropyridine (M19)

[0123]

[0124] The yield was 57% when isopropyl iodide was replaced by ethyl iodide and the synthesis method of compound M1 was referred to. 1 H NMR (300MHz, Chloroform-d) δ7.28 (dd, J=2.6, 1.7Hz, 1H), 7.19 (dd, J=8.5, 2.6Hz, 1H), 4.03 (q, J=7.2Hz, 2H), 1.38 (t, J=7.2Hz, 3H).

[0125] (20) Synthesis of 2-chloro-4-(6-ethoxy-5-fluoropyridin-3-yl)-5-fluoropyrimidine (M20)

[0126]

[0127] The yield of the synthetic method of reference compound M14 was 67%. 1H NMR (400MHz, Chloroform-d) δ8.77(dd,J=2.0,1.0Hz,1H),8.53(d,J=3.1Hz,1H),8.19(dd,J=11.0,2.0Hz,1H),4.57(q,J=7.1Hz,2H),1.49(t,J=7.1Hz,3H).

[0128] (21) Synthesis of 5-bromo-3-fluoro-N,N-dimethylpyridin-2-amine (M21)

[0129]

[0130] 5-Bromo-2,3-difluoropyridine (2.2 mL, 20 mmol), N,N-dimethylformamide (3.1 mL, 40 mmol), and sodium tert-butoxide (5.8 g, 60 mmol) were added to an eggplant-shaped bottle, and 80 mL of water was added. The mixture was heated to 140°C and reacted for 12 h. After the reaction was completed, the mixture was cooled to 25°C and purified by rapid silica gel column to obtain compound M21 (2.5 g, yield was 57%). The yield was 57%. 1 H NMR (400MHz, Chloroform-d) δ7.98 (dd, J=2.0, 1.2Hz, 1H), 7.29 (dd, J=12.6, 2.0Hz, 1H), 3.06 (d, J=2.4Hz, 6H).

[0131] (22) Synthesis of 5-(2-chloro-5-fluoropyrimidin-4-yl)-3-fluoro-N,N-dimethylpyridin-2-amine (M22).

[0132]

[0133] The yield of the synthetic method of reference compound M14 was 67%. 1 H NMR (400MHz, Chloroform-d) δ8.81-8.80 (m, 1H), 8.41 (d, J = 3.4Hz, 1H), 8.04 (dd, J = 15.4, 2.0Hz, 1H), 3.28 (d, J = 2.6Hz, 6H).

[0134] (23) Synthesis of 5-bromo-3-fluoro-N-isopropylpyridin-2-amine (M23)

[0135]

[0136] 5-Bromo-2,3-difluoropyridine (3.88 mg, 20 mmol), isopropylamine (2.1 mL, 24 mmol), and N,N-diisopropylethylamine (10.5 mL, 60 mmol) were added to an eggplant-shaped flask, and 80 mL of DMSO was added. The mixture was heated to 80°C and reacted for 2 h. After the reaction was completed, compound M21 (2.80 g, yield was 60%) was obtained by rapid silica gel column purification. 1 H NMR (400MHz, Chloroform-d) δ7.94(d,J=2.0Hz,1H),7.25(dd,J=10.3,2.0Hz,1H),4.43(d,J=7.5Hz,1H),4.24-4.17(m,1H),1.25(d,J=6.4Hz,6H).

[0137] (24) Synthesis of 5-(2-chloro-5-fluoropyrimidin-4-yl)-3-fluoro-N-isopropylpyridin-2-amine (M24).

[0138]

[0139] The yield of the synthetic method of reference compound M14 was 67%. 1 H NMR(400MHz, DMSO-d6)δ8.82(d,J=3.7Hz,1H),8.62-8.61(m,1H),7.88(dd,J=12.7 ,1.9Hz,1H),7.46(dd,J=8.0,1.9Hz,1H),4.39-4.30(m,1H),1.22(d,J=6.5Hz,6H).

[0140] 2. Synthesis of Compounds S-1-S-17

[0141] Example 1: Synthesis of 4-(5-fluoro-6-isopropoxypyridin-3-yl)-N-(1-(methylsulfonyl)piperidin-4-yl)pyrimidin-2-amine (S-1)

[0142]

[0143] Compound M2 (535 mg, 2 mmol) and 1-methylsulfonyl-4-aminopiperidine (445 mg, 2.5 mmol) were dissolved in dioxane (6 mL), and then Pd2(dba)3 (92 mg, 0.1 mmol), Xantphos (116 mg, 0.2 mmol), cesium carbonate (1.3 g, 4 mmol) were added, argon was replaced three times, heated to 105 ° C, and reacted for 12 h. Cooling, filtration, concentration, column chromatography, and compound S-1 (393 mg, 48% yield) were obtained as a white solid. 1H NMR (400MHz, DMSO-d6) δ8.79-8.74(m,1H),8.36(d,J=5.2Hz,1H),8.31-8.25(m,1H),7.31(d,J=7.9Hz,1H),7.20(d,J=5.2Hz,1H),5.43-5.3 7(m,1H),4.04-3.92(m,1H),3.57-3.52(m,2H),2.97-2.90(m,2H),2.8 9(s,3H),2.02-1.97(m,2H),1.62-1.53(m,2H),1.36(d,J=6.2Hz,6H).

[0144] Example 2: Synthesis of 4-(5-fluoro-6-isopropoxypyridin-3-yl)-5-methoxy-N-(1-(methylsulfonyl)piperidin-4-yl)pyrimidin-2-amine (S-2)

[0145]

[0146] The synthetic method of reference example 1 has a yield of 45%, and the product is a white solid. 1 H NMR (400MHz, DMSO-d6) δ8.73(d,J=2.0Hz,1H),8.33(s,1H),8.21(dd,J=12.1,2.0Hz,1H),6.95(d,J=7.8Hz,1H),5.43-5.37(m,1H), 3.89-3.83(m,4H),3.55-3.50(m,2H),2.94-2.90(m,2H),2.88(s,3H),2.01-1.96(m,2H),1.60-1.50(m,2H),1.36(d,J=6.2Hz,6H).

[0147] Example 3: Synthesis of 5-fluoro-4-(5-fluoro-6-isopropoxypyridin-3-yl)-N-(1-(methylsulfonyl)piperidin-4-yl)pyrimidin-2-amine (S-3)

[0148]

[0149] The synthetic method of reference example 1 has a yield of 43% and is a white solid. 1H NMR(400MHz,Chloroform-d)δ8.69(s,1H),8.22(d,J=3.5Hz,1H),8.06(d,J=11.3Hz,1H),5.52-5.45(m,1H),5.06(d,J=7.7Hz,1H), 3.98-3.95(m,1H),3.81-3.76(m,2H),2.98-2.92(m,2H),2.83(s,3H),2.23-2.18(m,2H),1.71-1.62(m,3H),1.44(d,J=6.2Hz,6H).

[0150] Example 4: Synthesis of N-(1-(ethylsulfonyl)piperidin-4-yl)-5-fluoro-4-(5-fluoro-6-isopropoxypyridin-3-yl)pyrimidin-2-amine (S-4)

[0151]

[0152] Compound M6 (280 mg, 0.8 mmol) and triethylamine (243 mg, 2.4 mmol) were dissolved in dichloromethane (5 mL), placed in an ice bath, ethylsulfonyl chloride (154 mg, 1.2 mmol) was added dropwise, the mixture was returned to room temperature, stirred for 2 h, concentrated and purified by column chromatography to obtain compound S-4 (300 mg, 85% yield) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ8.61(s,1H),8.46(d,J=3.6Hz,1H),8.15(d,J=11.3Hz,1H),7.40(d,J=7.8Hz,1H),5.45-5.39(m,1H),3.95-3 .85(m,1H),3.62-3.57(m,2H),3.09-2.96(m,4H),1.99-1.94(m,2H),1.58-1.49(m,2H),1.37(d,J=6.2Hz,6H),1.23(t,J=7.4Hz,3H).

[0153] Example 5: Synthesis of 5-fluoro-4-(5-fluoro-6-isopropoxypyridin-3-yl)-N-(1-(isopropylsulfonyl)piperidin-4-yl)pyrimidin-2-amine (S-5)

[0154]

[0155] The synthetic method of reference example 4 has a yield of 80% and is a white solid. 1H NMR (400MHz, DMSO-d6) δ8.61(s,1H),8.46(d,J=3.6Hz,1H),8.15(d,J=11.6Hz,1H),7.40(d,J= 7.9Hz,1H),5.45-5.39(m,1H),3.96-3.87(m,1H),3.67-3.62(m,2H),3.32-3.27(m,1H),3.11-

[0156] 3.02(m,2H),1.96-1.91(m,2H),1.51-1.46(m,2H),1.37(d,J=6.2Hz,6H),1.23(d,J=6.5Hz,6H).

[0157] Example 6: Synthesis of N-(1-(cyclopropylsulfonyl)piperidin-4-yl)-5-fluoro-4-(5-fluoro-6-isopropoxypyridin-3-yl)pyrimidin-2-amine (S-6)

[0158]

[0159] The synthetic method of reference example 4 has a yield of 82% and is a white solid. 1 H NMR (400MHz, DMSO-d6) δ8.61(s,1H),8.47(d,J=3.6Hz,1H),8.16(d,J=11.6Hz,1H),7.40(d,J=7.7Hz,1H),5.45-5.39(m,1H),3.94-3.85(m,1 H),3.62-3.57(m,2H),3.05-2.97(m,2H),2.02-1.95(m,3H),1.60-1.5 3(m,2H),1.37(d,J=6.2Hz,6H),1.01-0.98(m,2H),0.95-0.92(m,2H).

[0160] Example 7: Synthesis of 5-fluoro-4-(5-fluoro-6-isopropoxypyridin-3-yl)-N-(1-((2-methoxyethyl)sulfonyl)piperidin-4-yl)pyrimidin-2-amine (S-7)

[0161]

[0162] The synthetic method of reference example 4 has a yield of 86%, and the product is a white solid. 1H NMR (400MHz, DMSO-d6) δ8.61 (s, 1H), 8.47 (d, J = 3.5Hz, 1H), 8.15 (d, J = 11. 6Hz,1H),7.40(d,J=7.8Hz,1H),5.45-5.39(m,1H),3.91-3.83(m,1H),3.67 (t,J=6.1Hz,2H),3.60-3.54(m,2H),3.32-3.30(m,2H),3.28(s,3H),2.99 -2.92(m,2H),1.99-1.94(m,2H),1.57-1.48(m,2H),1.37(d,J=6.2Hz,6H).

[0163] Example 8: Synthesis of 5-fluoro-4-(5-fluoro-6-isopropoxypyridin-3-yl)-N-(1-(propanesulfonyl)piperidin-4-yl)pyrimidin-2-amine (S-8)

[0164]

[0165] The synthetic method of reference example 4 has a yield of 89% and is a white solid. 1 H NMR (400MHz, DMSO-d6) δ8.62(s,1H),8.47(d,J=3.5Hz,1H),8.15(d,J=11.6Hz,1H),7.40(d,J= 7.8Hz,1H),5.45-5.39(m,1H),3.94-3.85(m,1H),3.61-3.56(m,2H),3.03-2.96(m,4H),1.99-

[0166] 1.94(m,2H),1.74-1.68(m,2H),1.57-1.50(m,2H),1.37(d,J=6.1Hz,6H),1.00(t,J=7.4Hz,3H).

[0167] Example 9: Synthesis of 5-fluoro-4-(6-isopropoxypyridin-3-yl)-N-(1-(methylsulfonyl)piperidin-4-yl)pyrimidin-2-amine (S-9)

[0168]

[0169] The synthetic method of reference example 1 has a yield of 47% and is a white solid. 1H NMR (400MHz, DMSO-d6) δ8.81-8.78(m,1H),8.43(d,J=3.7Hz,1H),8.25(d,J=8.7Hz,1H),7.34(d,J=7.8Hz,1H),6.91(dd,J=8.8,0.7Hz,1H),5.37 -5.30(m,1H),3.91-3.83(m,1H),3.56-3.51(m,2H),2.94-2.89(m,2H),2 .88(s,3H),2.01-1.96(m,2H),1.61-1.51(m,2H),1.33(d,J=6.2Hz,6H).

[0170] Example 10: Synthesis of 4-(6-ethoxypyridin-3-yl)-5-fluoro-N-(1-(methylsulfonyl)piperidin-4-yl)pyrimidin-2-amine (S-10)

[0171]

[0172] The synthetic method of reference example 1 has a yield of 42% and is a white solid. 1 H NMR(400MHz,Chloroform-d)δ8.91(d,J=2.4Hz,1H),8.28-8.25(m,1H),8.20(d,J=3.6Hz,1H),6.82(dd,J=8.8,0.7Hz,1H),5.02(d,J=7.6Hz,1H),4.4 4(q,J=7.1Hz,2H),4.01-3.92(m,1H),3.80-3.75(m,2H),2.98-2.91(m,2H) ,2.82(s,3H),2.23-2.18(m,2H),1.70-1.62(m,2H),1.43(t,J=7.1Hz,3H).

[0173] Example 11: Synthesis of 5-fluoro-4-(6-isopropoxy-5-methylpyridin-3-yl)-N-(1-(methylsulfonyl)piperidin-4-yl)pyrimidin-2-amine (S-11)

[0174]

[0175] The synthetic method of reference example 1 has a yield of 43% and is a white solid. 1H NMR (400MHz, Chloroform-d) δ8.73 (s, 1H), 8.19 (d, J = 3.7Hz, 1H), 8.06-8.05 (m, 1H), 5.44-5.37 (m, 1H), 5.01 (d, J = 7.7Hz, 1H), 4.00-3. 93(m,1H),3.79-3.75(m,2H),2.97-2.91(m,2H),2.82(s,3H),2.23(s,3H),2.21-2.18(m,2H),1.71-1.62(m,2H),1.39(d,J=6.2Hz,6H).

[0176] Example 12: Synthesis of 5-fluoro-4-(6-isopropoxy-2-methylpyridin-3-yl)-N-(1-(methylsulfonyl)piperidin-4-yl)pyrimidin-2-amine (S-12)

[0177]

[0178] The synthetic method of reference example 1 has a yield of 50% and is a white solid. 1 H NMR(400MHz,Chloroform-d)δ8.20(d,J=2.1Hz,1H),7.59(dd,J=8.5,1.2Hz,1H),6.59(d,J=8.5Hz,1H),5.40-5.34(m,1H),5.03(d,J=7.6Hz,1H),3.9 7-3.88(m,1H),3.78-3.73(m,2H),2.95-2.88(m,2H),2.80(s,3H),2.45(d, J=1.9Hz,3H),2.20-2.16(m,2H),1.69-1.62(m,2H),1.36(d,J=6.2Hz,6H).

[0179] Example 13: Synthesis of 5-(5-fluoro-2-((1-(methylsulfonyl)piperidin-4-yl)amino)pyrimidin-4-yl)-2-isopropoxynicotinic acid (S-13)

[0180] 1. Synthesis of 5-(5-fluoro-2-((1-(methylsulfonyl)piperidin-4-yl)amino)pyrimidin-4-yl)-2-isopropoxynicotinate (S-13-1)

[0181]

[0182] The synthetic method of reference example 1 has a yield of 42% and is a white solid. 1H NMR(400MHz,Chloroform-d)δ9.01(d,J=2.5Hz,1H),8.82(d,J=2.5Hz,1H),8.23(d,J=3 .5Hz,1H),5.55-5.49(m,1H),5.07(d,J=7.6Hz,1H),4.01-3.95(m,1H),3.93(s,3H),3.8 0-3.75 (m, 2H), 2.99-2.92 (m, 2H), 2.82 (s, 3H), 2.23-2.19 (m, 2H), 1.71-1.64 (m, 2H), 1.43 (d, J = 6.2 Hz, 6H). 2. Synthesis of 5-(5-fluoro-2-((1-(methylsulfonyl)piperidin-4-yl)amino)pyrimidin-4-yl)-2-isopropoxynicotinic acid (S-13)

[0183]

[0184] Compound S-13-1 (374 mg, 0.8 mmol) was dissolved in 5 mL of methanol, placed in an ice bath, and 800 μL of 1 M lithium hydroxide aqueous solution was slowly added dropwise, reacted for 15 min, heated to 60° C., and reacted for 12 h. The mixture was cooled, filtered, concentrated, and column chromatographed to obtain compound S-13 (327 mg, 90% yield) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ13.10(s,1H),8.93(s,1H),8.68(d,J=2.5Hz,1H),8.46(d,J=3.6Hz,1H),7.41(d,J=7.7Hz,1H),5.48-5.42(m,1 H),3.90-3.83(m,1H),3.55-3.51(m,2H),2.93-2.90(m,2H),2.88(s,3H),2.01-1.98(m,2H),1.61-1.52(m,2H),1.35(d,J=6.2Hz,6H).

[0185] Example 14: Synthesis of 5-fluoro-4-(5-fluoro-6-methoxypyridin-3-yl)-N-(1-(methylsulfonyl)piperidin-4-yl)pyrimidin-2-amine (S-14)

[0186]

[0187] The synthetic method of reference example 1 has a yield of 49% and is a white solid. 1H NMR (400MHz, DMSO-d6) δ8.63(s,1H),8.48(d,J=3.6Hz,1H),8.17(d,J=11.5Hz,1H),7.41(d,J=7.8Hz,1H),4.03(s ,3H),3.93-3.83(m,1H),3.56-3.52(m,2H),2.94-2.90(m,2H),2.88(s,3H),2.01-1.97(m,2H),1.61-1.51(m,2H).

[0188] Example 15: Synthesis of 4-(6-ethoxy-5-fluoropyridin-3-yl)-5-fluoro-N-(1-(methylsulfonyl)piperidin-4-yl)pyrimidin-2-amine (S-15)

[0189]

[0190] The synthetic method of reference example 1 has a yield of 47% and is a white solid. 1 H NMR (400MHz, DMSO-d6) δ8.61(s,1H),8.47(d,J=3.6Hz,1H),8.16(dd,J=11.8,1.9Hz,1H),7.40(d,J=7.8Hz,1H),4.49(q,J=7.0 Hz,2H),3.92-3.84(m,1H),3.56-3.51(m,2H),2.94-2.86(m,5H),2.01-1.97(m,2H),1.61-1.52(m,2H),1.39(t,J=7.1Hz,3H).

[0191] Example 16: Synthesis of 4-(6-(dimethylamino)-5-fluoropyridin-3-yl)-5-fluoro-N-(1-(methylsulfonyl)piperidin-4-yl)pyrimidin-2-amine (S-16)

[0192]

[0193] The synthetic method of reference example 1 has a yield of 45%, and the product is a white solid. 1H NMR (400MHz, DMSO-d6) δ8.63-8.61(m,1H),8.37(d,J=3.9Hz,1H),7.92(dd,J=15.6,1.9Hz,1H),7.27(d,J=7.8Hz,1H),3.90- 3.83(m,1H),3.57-3.52(m,2H),3.16(d,J=2.6Hz,6H),2.94-2.90(m,2H),2.89(s,3H),2.01-1.96(m,2H),1.62-1.54(m,2H).

[0194] Example 17: Synthesis of 5-fluoro-4-(5-fluoro-6-(isopropylamino)pyridin-3-yl)-N-(1-(methylsulfonyl)piperidin-4-yl)pyrimidin-2-amine (S-17)

[0195]

[0196] The synthetic method of reference example 1 has a yield of 46% and is a white solid. 1 H NMR (400MHz, DMSO-d6) δ8.57(s,1H),8.35(d,J=4.0Hz,1H),7.87(dd,J=12.9,1.9Hz,1H),7.22(d,J=7.8Hz,1H),7.13-7.11(m,1H),4.35-4. 27(m,1H),3.90-3.82(m,1H),3.56-3.52(m,2H),2.94-2.90(m,2H),2. 88(s,3H),2.00-1.96(m,2H),1.60-1.50(m,2H),1.21(d,J=6.5Hz,6H)

[0197] 3. Biological Evaluation Experiment

[0198] (1) CDK2 kinase activity analysis method

[0199] a) Prepare ATP / substrate solution and CDK2 kinase solution using kinase reaction buffer.

[0200] b) Transfer compound dilutions to a 384-well plate; centrifuge and add CDK2 kinase solution to the 384-well plate, continue centrifugation for 1 minute, and then incubate at 25°C for 10 minutes.

[0201] c) Add ATP / substrate solution to the 384-well plate, centrifuge for 1 minute, and then incubate at 25°C for 60 minutes.

[0202] d) Add ADP-Glo ​​to the 384-well plate, centrifuge for 1 minute, and then incubate at 25°C for 40 minutes.

[0203] e) The reaction solution was transferred to another 384-well plate, centrifuged at 1000 rpm for 1 minute, and then incubated at 25°C for 40 minutes.

[0204] f) Read the luminescent signal with a multifunctional microplate reader and calculate the inhibition rate and IC of the compound on CDK2 kinase 50 value.

[0205] (2) CDK6 kinase activity analysis method

[0206] a) Prepare ATP / substrate solution and CDK6 kinase solution using kinase reaction buffer.

[0207] b) Transfer compound dilutions to a 384-well plate; centrifuge and add CDK6 kinase solution to the 384-well plate, continue centrifugation for 1 minute, and then incubate at 25°C for 10 minutes.

[0208] c) Add ATP / substrate solution to the 384-well plate, centrifuge for 1 minute, and then incubate at 25°C for 60 minutes.

[0209] d) Add ADP-Glo ​​to the 384-well plate, centrifuge for 1 minute, and then incubate at 25°C for 40 minutes.

[0210] e) The reaction solution was transferred to another 384-well plate, centrifuged at 1000 rpm for 1 minute, and then incubated at 25°C for 40 minutes.

[0211] f) Read the luminescent signal with a multifunctional microplate reader and calculate the inhibition rate and IC of the compound on CDK6 kinase 50 value.

[0212] Table 1 CDK kinase inhibitory activity of the compounds of the present invention

[0213]

[0214]

[0215] A:IC 50 ≤100nM; B:100nM <IC 50 ≤200nM; C:200nM <IC 50 ≤1000nM; “-”: not tested.

Claims

1. A compound represented by formula (S) or a pharmaceutically acceptable salt thereof, characterized in that: in, R1 is selected from OR6 or NR7R8, wherein R6 is selected from C1-C3 alkyl, and R7 and R8 are selected from hydrogen or C1-C3 alkyl; R2 is selected from hydrogen, halogen, -COOH or C1-C3 alkyl; R3 is selected from hydrogen or C1-C3 alkyl; R4 is selected from hydrogen, halogen or -OC1-C3 alkoxy; R5 is selected from -CH2CH2OCH3, -C1-C5 alkyl or cyclopropyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The R1 is selected from OR6 or NR7R8, wherein R6 is selected from methyl, ethyl, isopropyl, R7 is selected from hydrogen or methyl, and R8 is selected from methyl or isopropyl; Said R2 is selected from hydrogen, fluorine, -COOH, methyl or ethyl; Said R3 is selected from hydrogen, methyl or ethyl; Said R4 is selected from hydrogen, halogen or -OC1-C3 alkoxy; The R5 is selected from -CH2CH2OCH3, -C1-C5 alkyl or cyclopropyl.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The R1 is selected from OR6 or NR7R8, wherein R6 is selected from methyl, ethyl, isopropyl, R7 is selected from hydrogen or methyl, and R8 is selected from methyl or isopropyl; The R2 is selected from hydrogen, fluorine, -COOH, and methyl; Said R3 is selected from hydrogen and methyl; Said R4 is selected from hydrogen, fluorine or -OC1-C3 alkoxy; The R5 is selected from -CH2CH2OCH3, -C1-C5 alkyl or cyclopropyl.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The R1 is selected from OR6 or NR7R8, wherein R6 is selected from methyl, ethyl, isopropyl, R7 is selected from hydrogen or methyl, and R8 is selected from methyl or isopropyl; Said R2 is selected from hydrogen, fluorine, -COOH or methyl; Said R3 is selected from hydrogen or methyl; Said R4 is selected from hydrogen, fluorine or methoxy; The R5 is selected from -CH2CH2OCH3, -C1-C4 alkyl or cyclopropyl.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The R1 is selected from OR6 or NR7R8, wherein R6 is selected from methyl, ethyl, isopropyl, R7 is selected from hydrogen or methyl, and R8 is selected from methyl or isopropyl; Said R2 is selected from hydrogen, fluorine, -COOH or methyl; Said R3 is selected from hydrogen or methyl; Said R4 is selected from hydrogen, fluorine or methoxy; The R5 is selected from -CH2CH2OCH3, methyl, ethyl, n-propyl, isopropyl or cyclopropyl.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: Select any of the following compounds:

7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The pharmaceutically acceptable salt is an acid addition salt of the compound of formula (S), wherein the acid used for salt formation includes inorganic acids and organic acids, the inorganic acids include hydrochloric acid, sulfuric acid, phosphoric acid and methanesulfonic acid, and the organic acids include acetic acid, trichloroacetic acid, propionic acid, butyric acid, maleic acid, p-toluenesulfonic acid, malic acid, malonic acid, cinnamic acid, citric acid, fumaric acid, camphoric acid, digluconic acid, aspartic acid and tartaric acid.

8. A method for preparing the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof, characterized in that: Compound A and compound B are reacted with each other under the action of a palladium catalyst to prepare compound S; or compound C reacts with compound D under the action of a base to prepare compound S; R1 is selected from OR6 or NR7R8, wherein R6 is selected from C1-C3 alkyl, and R7 and R8 are selected from hydrogen or C1-C3 alkyl; R2 is selected from hydrogen, halogen, -COOH or C1-C3 alkyl; R3 is selected from hydrogen or C1-C3 alkyl; R4 is selected from hydrogen, halogen or -OC1-C3 alkoxy; R5 is selected from -CH2CH2OCH3, -C1-C5 alkyl or cyclopropyl.

9. Use of the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof in the preparation of a CDK2 or CDK6 kinase inhibitor.

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