Double-target PARP / CDK6 inhibitor as well as preparation method and medical application thereof

Abstract

The invention discloses a double-target PARP / CDK6 inhibitor and a preparation method and medical application thereof.The inhibitor comprises a compound with the structure shown in the general formula or pharmaceutically acceptable salt of the compound. The inhibitor can target PARP1 and CDK6 at the same time, can be used as a single therapeutic agent for tumors or can be combined with other anti-tumor drugs for use, and the application prospect is wide. Therefore, the effects of improving the curative effect of the existing anti-tumor drug and reducing the dosage and toxicity are achieved.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry technology and relates to the synthesis, preparation and use of new compounds, specifically dual-target PARP / CDK6 inhibitors, their preparation methods and pharmaceutical uses. Background Technology

[0002] Breast cancer has become one of the most common malignant tumors worldwide, and it is also the leading cause of cancer-related morbidity and mortality among women globally. Triple-negative breast cancer, in particular, has a poor prognosis, high metastasis rate, limited treatment options, and a high mortality rate. PARP inhibitors can induce synthetic lethality in cells with BRCA mutations in breast cancer, and therefore have been approved clinically for the treatment of BRCA-mutated breast cancer patients. However, their drug resistance and limited applicability severely restrict their clinical efficacy. Summary of the Invention

[0003] The purpose of this invention is to propose a new class of dual-target PARP / CDK6 inhibitors and their preparation method. Pharmacodynamic test results show that the compounds of this invention or their pharmaceutically acceptable salts can be used as single therapeutic agents for tumors.

[0004] The present invention discloses a dual-target PARP / CDK6 inhibitor, characterized in that it comprises a compound with the structure shown in the general formula or a pharmaceutically acceptable salt thereof:

[0005]

[0006] Wherein, R is NHCH2, NH(CH2)2, NHCH(CH3), N(CH3)CH2,

[0007] The preferred compounds of this invention are as follows:

[0008] (1) Nitrogen-(2-(4-((8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)-2-oxoethyl)-2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzamide.

[0009]

[0010] (2) Nitrogen-(3-(4-((8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)-3-oxopropyl)-2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzamide.

[0011]

[0012] (3) Nitrogen-(1-(4-((8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)-1-oxopropane-2-yl)-2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzamide.

[0013]

[0014] (4) Nitrogen-(2-(4-((8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)-2-oxoethyl)-2-fluoro-N-methyl-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzamide.

[0015]

[0016] (5) 8-Cyclopentyl-2-((1-(1-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)azacyclobutane-3-carbonyl)piperidin-4-yl)amino)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one.

[0017]

[0018] (6) 8-Cyclopentyl-2-((1-(1-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)pyrrolidine-3-carbonyl)piperidin-4-yl)amino)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one.

[0019]

[0020] (7) 8-Cyclopentyl-2-((1-(1-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)-3-methylpyrrolidine-3-carbonyl)piperidin-4-yl)amino)-5-methylpyrido[2,3-d]pyrimidine-7(8H)-one.

[0021]

[0022] (8) 8-Cyclopentyl-2-((1-(2-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)-2-azabicyclo[3.1.0]hexane-5-carbonyl)piperidin-4-yl)amino)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one.

[0023]

[0024] The present invention discloses a method for preparing a dual-target PARP / CDK6 inhibitor, comprising the following steps:

[0025] (1) 8-cyclopentyl-5-methyl-2-(piperidin-4-ylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (32.7 mg, 0.1 mmol) and the starting material of the dual-target pharmacophore linker, such as N-(tert-butoxycarbonyl)aminoacetic acid (17.5 mg, 0.1 mmol), were dissolved in dichloromethane. 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (38.0 mg, 0.1 mmol) and diisopropylethylamine (0.2 mL, 0.2 mmol) were added. After reacting for 2 h, the intermediate was obtained by column chromatography.

[0026] (2) Then the intermediate obtained in step (1) is dissolved in dichloromethane with trifluoroacetic acid (1.0 mL), reacted at room temperature for 2 h, and then evaporated to dryness for later use.

[0027] (3) The product of step (2) and 2-fluoro-5-[(4-oxo-3,4-dihydrophthalazin-1-yl)methyl]benzoic acid (26.9 mg, 0.1 mmol) were dissolved in dichloromethane, and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (38.0 mg, 0.1 mmol) and diisopropylethylamine (0.04 mL, 0.2 mmol) were added. After reacting for 2 h, compound 1 (38.5 mg) was separated by column chromatography.

[0028] The raw materials for the dual-target pharmacophore linker include, but are not limited to, N-(tert-butoxycarbonyl)aminoacetic acid, N-(tert-butoxycarbonyl)aminopropionic acid, Boc-DL-alanine, tert-butoxycarbonyl sarcosine, 1-Boc-azacyclobutane-3-carboxylic acid, 1-Boc-pyrrolidine-3-carboxylic acid, 3-methyl-1-[(2-methylpropane-2-yl)oxycarbonyl]pyrrolidine-3-carboxylic acid, or 3-[(2-methylpropane-2-yl)oxycarbonyl]-3-azabicyclo[3.1.0]hexane-1-carboxylic acid.

[0029] The synthesis methods of compounds 2-8 are the same as those of compound 1, except that the raw materials for the dual-target pharmacophore linker arm in step (1) are selected.

[0030]

[0031] The pharmaceutically acceptable salt described in this invention is an acid addition salt formed by the above-mentioned dual-target PARP / CDK6 inhibitor general formula compound and an acid, wherein the acid is selected from: hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, or ferulic acid.

[0032] A pharmaceutical composition comprising a compound of the above-described dual-target PARP / CDK6 inhibitor formula or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0033] The pharmaceutical composition can be prepared into any one of the following dosage forms: tablets, capsules, powders, syrups, liquids, suspensions, lyophilized powder for injection, or injections.

[0034] The use of the dual-target PARP / CDK6 inhibitor or a pharmaceutically acceptable salt thereof in the preparation of antitumor drugs.

[0035] The dual-target PARP / CDK6 inhibitor described in this invention is designed to increase the sensitivity of tumor cells to PARP inhibitors by inhibiting CDK6. Therefore, this inhibitor also exhibits good inhibitory effects on BRCA wild-type cells. It has the potential to address the resistance issues arising from PARP inhibitor monotherapy and broaden clinical indications to BRCA wild-type patients (including triple-negative breast cancer). Detailed Implementation

[0036] The present invention will be further illustrated by the following embodiments.

[0037] All chemical reagents used in this invention were purchased from domestic companies. The synthesis method is described in section 3. Example compound spectral data:

[0038] Example 1

[0039] Preparation of compound 1: N-(2-(4-((8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)-2-oxoethyl)-2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzamide.

[0040] (1) 8-cyclopentyl-5-methyl-2-(piperidin-4-ylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (32.7 mg) and N-(tert-butoxycarbonyl)aminoacetic acid (17.5 mg) were dissolved in dichloromethane, and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (38.0 mg) and diisopropylethylamine (0.2 mL) were added. After reacting for 2 h, the intermediate was obtained by column chromatography.

[0041] (2) Then the intermediate obtained in step (1) is dissolved in dichloromethane with trifluoroacetic acid (1.0 mL), reacted at room temperature for 2 h, and then evaporated to dryness for later use.

[0042] (3) The product of step (2) and 2-fluoro-5-[(4-oxo-3,4-dihydrophthalazin-1-yl)methyl]benzoic acid (26.9 mg) were dissolved in dichloromethane, and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (38.0 mg) and diisopropylethylamine (0.2 mL) were added. After reacting for 2 h, compound 1 (38.5 mg) was separated by column chromatography.

[0043] Results: Yield 58%; 1 H NMR (500MHz, CDCl3) δ (ppm): 10.13 (s, 1H), 8.53 (s, 1H), 8.45 (m, 1H), 8.05 (m, 1H),7.92(m,1H),7.76(m,4H),7.37(m,1H),7.09(m,1H),6.22(s,1H),5.82(t, J=9.0Hz,1H),4.31(m,5H),3.85(m,1H),3.19(t,J=12.5Hz,1H),2.80(m,1H), 2.38(m,2H),2.34(s,3H),2.21(m,2H),2.01(m,2H),1.85(m,2H),1.67(m,5H); 13 CNMR (125MHz, CDCl3)δ (ppm): 13 C NMR (125MHz, CHCl3) δ (ppm): 166.0, 164.0, 163.5, 162.9, 160.7, 158.8, 156.4, 155.5, 145.2, 134.2 (C, d, J C-C-C-C-F =2.5Hz), 133.7, 133.2(C,d,J) C-C-C-F =7.5Hz),131.7,131.6,129.5,128.3,127.2,125.1,121.3(C,d,J C-C-F =15.0Hz), 116.8(C,d,J) C-C-F =20.0Hz),48.2,43.8,42.0,40.1,37.8,35.8,32.6,29.7,28.0,18.6,17.2; ESI-MSm / z:665.37[M+H] + .

[0044] Example 2

[0045] Preparation of compound 2: N-(3-(4-((8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)-3-oxopropyl)-2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzamide.

[0046] Compound 2 was prepared using the same synthetic method as compound 1, except that in the first step of the reaction, N-(tert-butoxycarbonyl)aminoacetic acid was replaced with N-(tert-butoxycarbonyl)aminopropionic acid.

[0047] Results: Yield: 52%; 1 H NMR (500MHz, CDCl3) δ (ppm): 10.66 (s, 1H), 8.54 (s, 1H), 8.45 (d, J = 7.5Hz, 1H), 8.02 (d, J = 6.0Hz, 1 H),7.75(m,1H),7.58(s,1H),7.34(s,1H),7.04(t,J=9.0Hz,1H),6.21(s,1H),5.81(t,J=9.0Hz,1H ),4.53(m,1H),4.30(s,2H),4.10(m,1H),3.85(m,2H),3.19(t,J=12.5Hz,1H),2.89(m,1H),2.68( m,2H),2.38(m,2H),2.34(s,3H),2.11(m,2H),1.99(m,2H),1.82(m,2H),1.67(m,3H),1.45(m,2H); 13 CNMR (125MHz, CDCl3)δ (ppm): 13 C NMR (125MHz, CHCl3) δ (ppm): 169.9, 163.8, 163.2, 160.4, 160.3, 160.1, 158.8, 156.3, 155.9, 145.7, 134.2 (C, d, J C-C-C-C-F =2.5Hz), 133.7, 133.2(C,d,J) C-C-C-F =7.5Hz),131.7,131.6,129.6,128.3,127.2,125.1,121.3(C,d,J C-C-F =15.0Hz), 116.6(C,d,J) C-C-F =20.0Hz),48.3,44.2,40.5,37.8,35.8,32.6,29.7,28.0,25.6,17.2; ESI-MSm / z:679.41[M+H] +; HPLC purity 99.61% (MeOH / H2O=75 / 25; R t =4.870 min).

[0048] Example 3

[0049] Preparation of compound 3: N-(1-(4-((8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)-1-oxopropane-2-yl)-2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzamide.

[0050] Compound 3 was prepared using the same synthetic method as compound 1, except that in the first step of the reaction, N-(tert-butoxycarbonyl)aminoacetic acid was replaced with Boc-DL-alanine.

[0051] Results: Yield: 54%; 1 H NMR (500MHz, CDCl3) δ (ppm): 9.98 (s, 1H), 8.51 (m, 1H), 8.46 (m, 1H), 8.03 (m, 1H), 7 .92(m,1H),7.76(m,3H),7.35(m,1H),7.08(m,1H),6.23(m,1H),5.82(m,1H),5.13 (m,1H),4.30(s,2H),4.17(m,1H),4.00(m,1H),3.31(m,1H),2.90(s,1H),2.58(m, 1H),2.37(m,2H),2.34(s,3H),2.15(m,2H),2.02(m,3H),1.84(m,2H),1.42(s,3H); 13 CNMR (125MHz, CDCl3)δ (ppm): 13 C NMR (125MHz, CHCl3) δ (ppm): 170.4, 164.0, 162.2, 160.0, 157.0, 145.7, 135.2, 134.2 (C, d, J C-C-C-C-F =2.5Hz), 133.7, 133.2(C,d,J) C-C-C-F =7.5Hz),131.7,131.6,129.5,128.4,127.2,125.1,124.1,123.5,121.3(C,d,J C-C-F =15.0Hz), 116.6(C,d,J) C-C-F=20.0Hz),115.9,48.4,46.0,37.8,31.6,31.4,30.2,29.6,28.1,19.2,18.9,17.2; ESI-MSm / z:679.32[M+H] + ; HPLC purity 98.82% (MeOH / H2O=75 / 25; R t =5.726min).

[0052] Example 4

[0053] Preparation of compound 4: N-(2-(4-((8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)-2-oxoethyl)-2-fluoro-N-methyl-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzamide.

[0054] Compound 4 was prepared using the same synthetic method as compound 1, except that in the first step of the reaction, N-(tert-butoxycarbonyl)aminoacetic acid was replaced with tert-butoxycarbonylsarcosine.

[0055] Results: Yield: 50%; 1 H NMR (500MHz, CDCl3) δ (ppm): 10.66 (s, 1H), 8.65 (m, 1H), 8.46 (m, 1H), 7.76 (m, 2H) ,7.70(m,1H),7.30(m,2H),7.04(t,J=9.0Hz,1H),5.94(m,1H),4.31(m,1H),4.27 (s,2H),4.10(m,1H),3.69(m,2H),3.35(d,J=7.5Hz,1H),2.90(s,1H),2.58(m,1H ),2.57(s,3H),2.33(m,2H),2.21(m,2H),2.07(m,2H),2.02(m,3H),1.84(m,2H); 13 CNMR (125MHz, CDCl3)δ (ppm): 13 C NMR (125MHz, CHCl3) δ (ppm): 169.4, 164.0, 163.1, 162.9, 159.9, 154.4, 145.2, 134.2 (C, d, J C-C-C-C-F =2.5Hz), 133.7, 133.4 (C,d,J) C-C-C-F =7.5Hz),131.7,131.6,129.5,128.3,127.3,125.1,121.3(C,d,J C-C-F=15.0Hz), 116.8(C,d,J) C-C-F =20.0Hz),53.0,48.0,44.4,37.7,31.6,30.0,28.1,25.5,17.4; ESI-MSm / z:679.30[M+H] + ; HPLC purity 99.35% (MeOH / H2O=75 / 25; R t =4.442 min).

[0056] Example 5

[0057] Preparation of compound 5: 8-cyclopentyl-2-((1-(1-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)azacyclobutane-3-carbonyl)piperidin-4-yl)amino)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one.

[0058] Compound 5 was prepared using the same synthetic method as compound 1, except that in the first step of the reaction, N-(tert-butoxycarbonyl)aminoacetic acid was replaced with 1-Boc-azacyclobutane-3-carboxylic acid.

[0059] Results: Yield: 48%; 1 H NMR (500MHz, CDCl3) δ (ppm): 10.44 (s, 1H), 8.53 (s, 1H), 8.45 (d, J = 7.5Hz, 1H) ,7.77(m,3H),7.51(s,1H),7.33(s,1H),6.21(s,1H),5.81(t,J=9.0Hz,1H),4 .39(m,3H),4.30(m,4H),4.21(m,3H),3.60(m,2H),3.18(m,1H),2.96(m,1H), 2.38(m,2H),2.34(s,3H),2.13(m,2H),2.00(m,2H),1.83(m,2H),1.46(m,3H); 13 CNMR(125MHz, CDCl3)δ(ppm):169.0,166.3,166.2,163.8,160.3,160.1,156.3,155.9,145.6,134.2(C,d,J C-C-C-C-F =2.5Hz), 133.7, 132.5(C,d,J) C-C-C-F =7.5Hz),131.6,130.2,129.5,128.3,127.2,125.1,121.8(C,d,J C-C-F =15.0Hz), 116.6(C,d,J) C-C-F=20.0Hz),51.1,48.3,37.7,31.4,29.7,28.1,25.6,18.6,17.2; ESI-MSm / z:691.40[M+H] + ; HPLC purity 99.73% (MeOH / H2O=75 / 25; R t =4.759min).

[0060] Example 6

[0061] Preparation of compound 6: 8-cyclopentyl-2-((1-(1-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)pyrrolidine-3-carbonyl)piperidin-4-yl)amino)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one.

[0062] Compound 6 was prepared using the same synthetic method as compound 1, except that in the first step of the reaction, N-(tert-butoxycarbonyl)aminoacetic acid was replaced with 1-Boc-pyrrolidine-3-carboxylic acid.

[0063] Results: Yield: 54%; 1 H NMR (500MHz, CDCl3) δ (ppm): 10.93 (s, 1H), 8.56 (m, 1H), 8.45 (m, 1H), 7.75 (m, 3H), 7.40 (m ,1H),7.27(m,1H),7.01(m,1H),6.22(s,1H),5.81(m,1H),4.47(s,1H),4.28(s,2H),4.13 (m,1H),3.98(m,2H),3.72(m,2H),3.47(m,1H),3.34(m,2H),2.95(m,1H),2.38(m,2H),2. 34(s,3H),2.20(m,1H),2.17(m,2H),2.03(m,4H),1.82(m,2H),1.67(m,2H),1.53(m,2H); 13 C NMR (125MHz, CDCl3) δ (ppm): 13 C NMR (125MHz, CHCl3) δ (ppm): 170.3, 169.7, 164.8, 163.7, 160.5, 158.3, 156.6, 156.3, 155.5, 145.6, 145.5, 134.1 (C, d, J C-C-C-C-F =2.5Hz),133.8,131.6,131.4(C,d,J C-C-C-F=7.5Hz),129.6,129.0,128.4,127.2,125.4,125.1(C,d,J C-C-F =15.0Hz), 118.3, 116.4 (C,d,J) C-C-F =20.0Hz),60.4,48.8,48.4,47.4,45.4,44.2,40.7,39.3,37.8,29.1,28.1,25.6,21.0,17.1,14.2; ESI-MSm / z:705.43[M+H] + ; HPLC purity 99.72% (MeOH / H2O=75 / 25; R t =4.553min).

[0064] Example 7

[0065] Preparation of compound 7: 8-cyclopentyl-2-((1-(1-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)-3-methylpyrrolidine-3-carbonyl)piperidin-4-yl)amino)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one.

[0066] Compound 7 was prepared using the same synthetic method as compound 1, except that in the first step of the reaction, N-(tert-butoxycarbonyl)aminoacetic acid was replaced with 3-methyl-1-[(2-methylpropane-2-yl)oxycarbonyl]pyrrolidine-3-carboxylic acid.

[0067] Results: Yield: 49%; 1 H NMR (500MHz, CDCl3) δ (ppm): 10.66 (s, 1H), 8.56 (t, J = 17.5Hz, 1H), 8.46 (m, 1H) ,7.77(m,3H),7.38(m,1H),7.33(m,1H),7.29(m,1H),7.03(m,1H),6.22(s,1H) ,5.81(m,1H),4.27(m,3H),3.76(m,2H),3.35(m,2H),3.07(m,1H),2.58(m,1H) ,2.38(m,2H),2.34(s,3H),2.15(m,2H),2.00(m,4H),1.83(m,3H),1.68(m,2H); 13 C NMR (125MHz, CDCl3) δ (ppm): 172.9, 165.2, 163.8, 160.3, 156.4, 155.9, 145.6, 134.2 (C, d, J C-C-C-C-F =2.5Hz), 133.7, 131.7(C,d,J)C-C-C-F =7.5Hz),129.5,129.2,128.9,128.4,127.2,125.1(C,d,J C-C-F =15.0Hz), 116.3(C,d,J) C-C-F =20.0Hz),49.1,48.4,47.7,44.5,37.7,37.4,32.3,29.7,28.0,22.1,17.2; ESI-MSm / z:719.47[M+H] + ; HPLC purity99.28% (MeOH / H2O=75 / 25; R t =4.888min).

[0068] Example 8

[0069] Preparation of compound 8: 8-cyclopentyl-2-((1-(2-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)-2-azabicyclo[3.1.0]hexane-5-carbonyl)piperidin-4-yl)amino)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one.

[0070] Compound 8 was prepared according to the synthetic method of compound 1, except that in the first step of the reaction, N-(tert-butoxycarbonyl)aminoacetic acid was replaced with 3-[(2-methylpropane-2-yl)oxycarbonyl]-3-azabicyclo[3.1.0]hexane-1-carboxylic acid.

[0071] Results: Yield: 48%; 1 H NMR (500MHz, CDCl3) δ (ppm): 10.32 (s, 1H), 8.55 (m, 1H), 8.45 (m, 1H), 7.77 (m, 2H),7.70(m,1H),7.29(m,2H),7.04(t,J=7.5Hz,1H),6.22(s,1H),5.82(m,1H ),4.36(m,1H),4.27(s,2H),4.12(m,1H),3.65(m,3H),3.34(t,J=9.0Hz,1H), 3.16(m,2H),2.39(m,2H),2.35(s,3H),2.16(m,1H),2.03(m,3H),1.83(m,2H); 13 C NMR (125MHz, CDCl3) δ (ppm): 168.4, 166.0, 163.8, 160.3, 156.4, 155.9, 145.5, 134.0 (C, d, J C-C-C-C-F =2.5Hz), 133.7, 131.7(C,d,J) C-C-C-F=7.5Hz),129.5,128.7,128.4,127.2,125.1(C,d,J C-C-F =15.0Hz), 116.3(C,d,J) C-C-F =20.0Hz),49.4,48.4,47.9,43.5,37.7,30.2,28.1,18.6,17.2; ESI-MSm / z:717.43[M+H] + ; HPLC purity 97.74% (MeOH / H2O=75 / 25; R t =4.956 min).

[0072] Example 9

[0073] Activity testing: The pharmacological experiments and results of some compounds in this invention are as follows:

[0074] PARP1 and CDK6 inhibitory activities; MDA-MB-231, MCF-7, and MCF-10A cell inhibitory activities:

[0075] Experimental methods: The in vitro inhibitory activity of the compounds against PARP1 was determined using the HT universal chemiluminescent PARP assay kit, with Olaparib as a positive control; the in vitro inhibitory activity of the compounds against CDK family kinases was determined using the ADP-Glo ​​kinase assay kit, with Palbociclib as a positive control; the inhibitory activity against MDA-MB-231 and MCF-7 cells was determined using the MTT assay to obtain the IC50. 50 Three subwells were set up for each compound, and each experiment was repeated three times. The experimental results are expressed as mean ± SEM.

[0076] Table 1 shows the inhibitory activity of the compounds of the present invention against PARP-1 and CDK6; and the inhibitory activity of the compounds of the present invention against MDA-MB-231 and MCF-7 cells.

[0077]

[0078] a NT = Not detected;

[0079] Cell viability was detected by MTT assay after 72 hours of treatment.

[0080] Table 2. Inhibitory activity of compound 6 of the present invention against CDK family kinases at 500 nM.

[0081]

[0082]

[0083] a average inhibition rate

[0084] Table 1 shows that compound 6 has good inhibitory effects on several tumor cells, even better than the combination of positive control drugs Olaparib and Palbociclib. At the same time, it has low toxicity to normal human breast cells MCF-10A and maintains good in vitro inhibitory activity against PARP1 and CDK6. Table 2 shows that compound 6 has much higher inhibitory activity against CDK4 / 6 kinase than other CDK kinases, and has good selective inhibition, showing potential as an anti-tumor drug.

Claims

1. A dual-target PARP / CDK6 inhibitor, characterized in that... Includes compounds with structures as shown in the general formula or their pharmaceutically acceptable salts: Wherein, R is NHCH2, NH(CH2)2, NHCH(CH3), N(CH3)CH2, 2. The dual-target PARP / CDK6 inhibitor according to claim 1, characterized in that... The dual inhibitor is one of the following compounds: (1) Nitrogen-(2-(4-((8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)-2-oxoethyl)-2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzamide: (2) Nitrogen-(3-(4-((8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)-3-oxopropyl)-2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzamide: (3) Nitrogen-(1-(4-((8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)-1-oxopropane-2-yl)-2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzamide: (4) N-(2-(4-((8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)-2-oxoethyl)-2-fluoro-N-methyl-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzamide: (5) 8-Cyclopentyl-2-((1-(1-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)azacyclobutane-3-carbonyl)piperidin-4-yl)amino)-5-methylpyridino[2,3-d]pyrimidin-7(8H)-one: (6) 8-Cyclopentyl-2-((1-(1-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)pyrrolidine-3-carbonyl)piperidin-4-yl)amino)-5-methylpyridino[2,3-d]pyrimidin-7(8H)-one: (7) 8-Cyclopentyl-2-((1-(1-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)-3-methylpyrrolidine-3-carbonyl)piperidin-4-yl)amino)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one: (8) 8-Cyclopentyl-2-((1-(2-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)-2-azabicyclo[3.1.0]hexane-5-carbonyl)piperidin-4-yl)amino)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one:

3. A method for preparing a dual-target PARP / CDK6 inhibitor as described in claim 1 or 2, characterized in that: Includes the following steps: (1) 32.7 mg and 0.1 mmol of compound 8-cyclopentyl-5-methyl-2-(piperidin-4-ylamino)pyrido[2,3-d]pyrimidin-7(8H)-one and 17.5 mg and 0.1 mmol of the dual-target pharmacophore linker were dissolved in dichloromethane, and 38.0 mg and 0.1 mmol of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and 0.2 mL and 0.2 mmol of diisopropylethylamine were added. After reacting for 2 h, the intermediate was obtained by column chromatography. (2) Then the intermediate obtained in step (1) was dissolved in 1.0 mL of trifluoroacetic acid in dichloromethane and reacted at room temperature for 2 h. The solution was then evaporated to dryness for later use. (3) Dissolve the product of step (2) and 26.9 mg and 0.1 mmol of 2-fluoro-5-[(4-oxo-3,4-dihydrophthalazin-1-yl)methyl]benzoic acid in dichloromethane, add 38.0 mg and 0.1 mmol of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and 0.04 mL and 0.2 mmol of diisopropylethylamine, react for 2 h, and then separate the compound by column chromatography. The raw materials for the dual-target pharmacophore linker include, but are not limited to, N-(tert-butoxycarbonyl)aminoacetic acid, N-(tert-butoxycarbonyl)aminopropionic acid, Boc-DL-alanine, tert-butoxycarbonyl sarcosine, 1-Boc-azacyclobutane-3-carboxylic acid, 1-Boc-pyrrolidine-3-carboxylic acid, 3-methyl-1-[(2-methylpropane-2-yl)oxycarbonyl]pyrrolidine-3-carboxylic acid, or 3-[(2-methylpropane-2-yl)oxycarbonyl]-3-azabicyclo[3.1.0]hexane-1-carboxylic acid.

4. A dual-target PARP / CDK6 inhibitor as described in claim 1 or 2, characterized in that... The pharmaceutically acceptable salt is an acid addition salt formed by the above-mentioned dual-target PARP / CDK6 inhibitor general formula compound and an acid, wherein the acid is selected from: hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, or ferulic acid.

5. A pharmaceutical composition, characterized in that... A compound containing the dual-target PARP / CDK6 inhibitor formula of claim 1 or 2, or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier thereof. The pharmaceutical composition can be prepared into any one of the following dosage forms: tablets, capsules, powders, syrups, liquids, suspensions, lyophilized powder for injection, or injections.

6. Use of a dual-target PARP / CDK6 inhibitor as described in claim 1 or 2, or a pharmaceutically acceptable salt thereof, in the preparation of an antitumor drug.

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