CDK12 / 13 inhibitor and application thereof

Abstract

The invention belongs to the field of medical chemistry, and particularly relates to a CDK12 / 13 inhibitor as shown in formula I or pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the compounds and application of the compounds or the composition to preparation of drugs for treating CDK12 / 13 mediated diseases, # imgabs0 #.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry, and specifically relates to a CDK12 / 13 inhibitor or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing these compounds, and the use of these compounds or compositions for preparing drugs for treating CDK12 / 13-mediated diseases. Background Art

[0002] Cyclin-dependent kinases (CDKs) are a family of protein kinases that catalyze serine / threonine phosphorylation and are closely related to the cell cycle and transcription. They have been confirmed to be oncogenic drivers. 21 subtypes of CDKs have been found. Different CDK subtypes have different functions, but they have highly similar ATP binding pockets. Therefore, although many reported pan-CDK inhibitors have significant anti-tumor effects, they also have problems such as large toxic side effects, which seriously limit their clinical application. Therefore, it is of great significance to develop selective CDK inhibitors.

[0003] Studies have shown that CDK12 and CDK13 play a series of important biological functions in gene transcription regulation, participation in pre-mRNA splicing, promotion of cell proliferation and differentiation, and DNA damage response. With the continuous deepening of the pathogenic mechanism and genomic research of breast cancer and other tumors, studies have found that CDK12 and CDK13 are abnormally expressed or mutated in a variety of cancers, which are closely related to their occurrence and development. CDK12 / 13 is expected to become a new biomarker and a new target for personalized treatment of breast cancer and prostate cancer patients. By inhibiting CDK12 / 13 kinases, the sensitivity of tumor cells to DNA damaging drugs can be enhanced, and genomic instability can be increased, thereby effectively inhibiting tumor growth. Therefore, the development of CDK12 / 13 inhibitors has become a potential new strategy for tumor treatment. Summary of the invention

[0004] One object of the present invention is to provide a CDK12 / 13 inhibitor represented by general formula I or a pharmaceutically acceptable salt thereof.

[0005] Another object of the present invention is to provide a method for preparing the compound of formula I or a pharmaceutically acceptable salt thereof.

[0006] Another object of the present invention is to provide a composition comprising a compound of the general formula I of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, as well as a composition comprising a compound of the general formula I of the present invention or a pharmaceutically acceptable salt thereof and another one or more drugs.

[0007] Another object of the present invention is to provide use of the compound of general formula I of the present invention or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating or preventing CDK12 / 13 related diseases.

[0008] To achieve the above purpose, the present invention provides the following technical solutions:

[0009] In a first aspect, the present invention provides a CDK12 / 13 inhibitor or an enantiomer or a pharmaceutically acceptable salt thereof as shown in Formula I:

[0010]

[0011] in

[0012] R1 is halogen;

[0013] R2 is selected from hydrogen, cycloalkyl, alkyl, and the alkyl may be substituted by alkyl.

[0014] In some preferred embodiments, R1 is selected from chlorine or bromine; preferably, R1 is bromine.

[0015] In some preferred embodiments, R2 is selected from hydrogen, cyclopropyl, N,N-dimethylamino.

[0016] The present invention provides the following specific compounds or pharmaceutically acceptable salts thereof:

[0017]

[0018] In another aspect, the present invention provides a method for preparing the compound of the general formula (I) of the present invention, comprising the following synthetic steps:

[0019] Step 1: The compound of formula (1) undergoes a nucleophilic substitution reaction with aniline to obtain a compound of formula (2);

[0020] Step 2: reacting the compound of formula (2) with the compound of formula (3) to obtain the compound of formula (4);

[0021] Step 3: condensing the compound of formula (6) with the compound of formula (7) to obtain the compound of formula (8);

[0022] Step 4: The compound of formula (8) reacts with the compound of formula (4) to obtain a compound of general formula (I). The reaction scheme is as follows:

[0023]

[0024] Wherein, R1 and R2 have the definitions given in the general formula (I).

[0025] In a third aspect, the present invention provides a pharmaceutical composition comprising the compound of the present invention or an enantiomer or a pharmaceutically acceptable salt thereof.

[0026] In some embodiments, the present invention provides a compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutical composition comprising the compound of the present invention or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutical composition is used for treating a CDK12 / 13-mediated disease.

[0027] In some embodiments, the present invention provides a pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0028] The compound of the present invention or its pharmaceutically acceptable salt can be mixed with a pharmaceutically acceptable carrier, diluent or excipient to prepare a pharmaceutical preparation, suitable for oral or parenteral administration. The method of administration includes, but is not limited to intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal and oral routes. The preparation can be administered by any route, such as by infusion or push injection, by the route of absorption through the epithelium or skin mucosa (such as oral mucosa or rectum, etc.). Administration can be systemic or local. Examples of oral administration preparations include solid or liquid dosage forms, specifically, tablets, pills, granules, powders, capsules, syrups, emulsions, suspensions, etc. The preparation can be prepared by methods known in the art, and includes carriers, diluents or excipients conventionally used in the field of pharmaceutical preparations.

[0029] In a fourth aspect, the present invention provides the use of the compound represented by formula I of the present invention or its enantiomer, pharmaceutically acceptable salt, or a pharmaceutical composition comprising the same in the preparation of a medicament for treating CDK12 / 13-mediated diseases.

[0030] In some preferred embodiments, the present invention provides the use of a compound represented by formula I of the present invention or its enantiomer, a pharmaceutically acceptable salt, or a pharmaceutical composition comprising the same in the preparation of a medicament for treating CDK12 / 13-mediated diseases, wherein the CDK12 / 13-mediated diseases include but are not limited to breast cancer, ovarian cancer, and prostate cancer.

[0031] Explanation of terms

[0032] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0033] The "halogen" in the present invention refers to fluorine, chlorine, bromine or iodine.

[0034] The "pharmaceutically acceptable salt" of the present invention refers to the salt of the compound of the present invention, which is safe and effective when used in mammals and has the desired biological activity.

[0035] The "pharmaceutical composition" of the present invention refers to a mixture comprising any one of the compounds of the present invention, including corresponding isomers, prodrugs, solvates, pharmaceutically acceptable salts or chemically protected forms thereof, and one or more pharmaceutically acceptable carriers and / or one or more other drugs. The purpose of the pharmaceutical composition is to facilitate the administration of the compound to an organism. The composition is generally used to prepare a drug for the treatment and / or prevention of a disease mediated by one or more kinases.

[0036] The "pharmaceutically acceptable carrier" of the present invention refers to a carrier that does not cause significant irritation to an organism and does not interfere with the biological activity and properties of the administered compound, including all solvents, diluents or other excipients, dispersants, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants, etc. Unless any conventional carrier medium is incompatible with the compound of the present invention. Some examples of pharmaceutically acceptable carriers include, but are not limited to, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, as well as cellulose and cellulose acetate; malt, gelatin, etc.

[0037] The "excipient" of the present invention refers to an inert substance added to a pharmaceutical composition to further facilitate administration of a compound. Excipients may include calcium carbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, vegetable oils, polyethylene glycols. DETAILED DESCRIPTION

[0038] The present invention is further described in detail below in conjunction with the examples, but the present invention is not limited to these examples. The materials used in the following examples are commercially available unless otherwise specified.

[0039] Example 1: Preparation of (E)-N-(3-(((5-chloro-4-(phenylamino)pyrimidin-2-yl)amino)piperidine-1-carbonyl)phenyl)-4-(dimethylamino)but-2-enamide (I-1)

[0040] Step 1: Synthesis of 2,5-dichloro-N-phenylpyrimidin-4-amine

[0041] Take 2,4,5-trichloropyrimidine (1 g, 5.46 mmol), aniline (0.56 g, 6.02 mmol) and N,N-diisopropylethylamine (DIEA) (0.7 g, 5.43 mmol) and dissolve them in 20 ml of isopropanol. Under N2 protection and an external temperature of 90°C, condense and reflux to react. After reacting for 2 hours, pour the reaction solution into a beaker, add water (H2O) and ethyl acetate (EA) to dilute (150 ml×3), wash with saturated sodium carbonate and saturated brine in turn, combine the organic phases, dry with anhydrous sodium sulfate powder, and spin-dry the organic layer to obtain 1.3 g of a yellow solid with a yield of 98%.

[0042] Step 2: Synthesis of tert-butyl 4-((5-chloro-4-(phenylamino)pyrimidin-2-yl)amino)piperidine-1-carboxylate

[0043] Weigh 2,5-dichloro-N-phenylpyrimidine-4-amine (1.3 g, 5.44 mmol), tert-butyl 4-aminopiperidine-1-carboxylate (1.08 g, 5.4 mmol), and N,N-diisopropylethylamine (DIEA) (0.70 g, 5.42 mmol) and dissolve them in N-methylpyrrolidone (NMP). After reflux reaction at 135°C for 3 h under N2 protection, add the reaction solution into a beaker, add water (H2O) and ethyl acetate (EA) for dilution (150 ml×3), wash with saturated sodium carbonate and saturated brine in turn, combine the organic phases, dry over anhydrous sodium sulfate, spin dry the organic layer to obtain a dark yellow liquid, remove the solvent under reduced pressure, and purify the residue by silica gel column chromatography to obtain 1.05 g of a white powdery solid compound with a yield of 48%.

[0044] Step 3: 5-Chloro-N 4 -phenyl-N 2 Synthesis of -(piperidin-4-yl)pyrimidine-2,4-diamine

[0045] 4-((5-chloro-4-(phenylamino)pyrimidin-2-yl)amino)piperidine-1-carboxylic acid tert-butyl ester (1.05 g, 2.61 mmol) was added to 20 ml of 1,4-dioxane solution, and then 0.33 ml of concentrated hydrochloric acid was added. The reaction was allowed to react for 3 h at room temperature under N2 protection. TLC monitoring showed that the reaction was complete. The reaction was cooled to room temperature, 100 ml of ethyl acetate (EA) was added to quench the reaction, and then saturated sodium carbonate (Na2CO3) solution was added to adjust the pH of the solution to alkaline and then transferred to a separatory funnel. After standing and stratifying, the organic layer was rotary evaporated, and the residue was purified by silica gel column chromatography to obtain 0.62 g of a light yellow solid compound with a yield of 77%.

[0046] Step 4: Synthesis of (E)-4-(3-(dimethylamino)-2-butenylamido)benzoic acid

[0047] Weigh N,N-dimethyl trans-crotonic acid (2g, 12.08mmol) and DMF (1ml) and dissolve them in acetonitrile (20ml). Under ice bath conditions, dissolve oxalyl chloride (1.6ml, 18.12mmol) in acetonitrile (8ml), add and stir to react. After the reaction is clarified, add to the reaction bottle of methyl 3-aminobenzoate (2g, 5.79mmol) and acetonitrile (15ml). After the addition is complete, move it into the reaction bottle at room temperature and stir to react for 1.5h. Pour the reaction solution into a beaker, add ethyl acetate (EA) and water to dilute, add saturated sodium carbonate solution to adjust the pH of the solution to alkaline, transfer it to a separatory funnel, separate the liquid, and obtain an oily product by column chromatography. Then add NaOH (0.22g, 1.5eq) dissolved in anhydrous ethanol (10ml) to the obtained product. The reaction mixture was condensed and refluxed at an external temperature of 50° C. After the reaction was complete, heating was stopped and HCl was added until the pH value was acidic. The resulting liquid was rotary evaporated to precipitate 1.24 g of a white solid with a yield of 62.3%.

[0048] Step 5: Synthesis of (E)-N-(3-(((5-chloro-4-(phenylamino)pyrimidin-2-yl)amino)piperidine-1-carbonyl)phenyl)-4-(dimethylamino)but-2-enamide (I-1)

[0049] Take compound 5-chloro-N 4 -phenyl-N 2 -(Piperidin-4-yl)pyrimidine-2,4-diamine (0.62g, 2.04mmol) and compound (E)-4-(3-(dimethylamino)-2-butenylamino)benzoic acid (0.73g, 2.57mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.59g, 3.09mmol), (1-hydroxybenzotriazole) (0.40g, 3.05mmol), N,N-diisopropylethylamine (0.49g, 3.80mmol) were dissolved in N,N-dimethylformamide in a 50ml reaction bottle, and reacted at room temperature for 1.5h. TLC monitoring showed that the reaction was complete. Saturated saline solution, water, and ethyl acetate were added. After standing and stratification, the organic layer was rotary evaporated, and the residue was purified by silica gel column chromatography to obtain 150mg of a white solid compound with a yield of 13.8%. 1HNMR(600MHz,DMSO-d6)δ11.11(s,1H),10.78(s,1H),8.62(s,1H),10.10(s,1H),8.31(s,1H),7. 86(s,1H),7.69(t,J=14.1Hz,1H),7.61(d,J=7.7Hz,1H),7.42(q,J=8.4,7.9Hz,3H),7.25(t,J=7. 4Hz,1H),7.09(d,J=7.5Hz,1H),6.90-6.84(m,1H),6.55(d,J=15.3Hz,1H),4.29(s,1H),4.01-3. 93(m,2H),3.77(s,2H),2.99-2.91(m,2H),2.75(d,J=3.7Hz,6H),1.84-1.89(m,2H),1.47(s,2H). 13 C NMR(151MHz,DMSO-d6)δ175.41,169.16,163.85,159.90,156.04,155.62,142.11,139.69,139.52,137.18,129.41,128.69, 126.26,123.59,122.37,121.87,120.42,117.76,106.46,101.84,60.12,55.33,46.70,45.58,41.14,34.78,31.68,29.90.

[0050] Example 5: Preparation of N-((1R,4R)-4-((5-chloro-4-(phenylamino)pyrimidin-2-yl)amino)cyclohexyl)-3-((E)-4-(dimethylamino)but-2-enamide)benzamide (1)

[0051] The operation was the same as in Example 1 to obtain 65 mg of white solid powder. 1H NMR (600MHz, DMSO-d6) δ10.20(s,1H),8.57(s,1H),8.24(s,1H),8.02(t,J=1.9Hz,1H),7.96(s,1 H),7.90–7.83(m,1H),7.79(d,J=8.0Hz,2H),7.52(d,J=7.7Hz,1H),7.39(t,J=7.9Hz,1H),7.33(t ,J=7.7Hz,2H),7.01-7.06(m,1H),6.96(s,1H),6.76-6.78(m,1H),6.24-6.28(m,1H),3.76(s,1H) ,3.51(s,1H),3.05-3.08(m,2H),2.19(s,6H),1.97(s,2H),1.91-1.83(m,2H),1.48-1.30(m,4H). 13 C NMR(151MHz,DMSO-d6)δ166.05,163.81,160.10,155.96,155.54,155.26,153.53,145.14,141.92,139.69,139.62,136.18,12 8.93,128.64,126.38,123.41,122.36,122.20,122.08,119.19,69.99,60.16,50.30,48.51,45.56,31.63,22.73,22.27,9.71.

[0052] Example 6: Preparation of 3-acrylamido-N-(4-((5-chloro-4-(phenylamino)pyrimidin-2-yl)amino)cyclohexyl)benzamide (2)

[0053] The same operation as in Example 1 was performed to obtain 77 mg of white solid powder. 1 H NMR(600MHz,DMSO-d6)δ10.41(s,1H),10.10(s,1H),8.44–8.35(m,1H),8.31–8.23(m, 1H),8.06(s,1H),7.93–7.86(m,1H),7.65(d,J=7.9Hz,2H),7.54(d,J=7.7Hz,1H),7.4 2-7.40(m,,3H),7.27(t,J=7.7Hz,1H),6.53-6.49(m,1H),6.31-6.28(m,1H),5.82–5. 71(m,2H),3.77(s,2H),1.96(d,J=11.4Hz,2H),1.90–1.76(m,2H),1.61–1.14(m,4H). 13C NMR(151MHz,DMSO-d6)δ166.02,163.76,157.67,152.39,142.37,139.49,137.13,136.03,132.29,128.98,128.79,1 27.46,126.41,124.88,124.72,122.54,122.29,119.21,105.49,103.38,66.82,50.79,47.96,31.05,30.88,27.86.

[0054] Example 7: Preparation of (E)-N-(3-((5-chloro-4-(phenylamino)pyrimidin-2-yl)amino)cyclohexyl)-3-(4-(dimethylamino)but-2-enamide)benzamide (3)

[0055] The operation was the same as in Example 1 to obtain 47 mg of a white solid powder, 1H NMR (600 MHz, Methanol-d4) δ 8.08 (t, J = 2.0 Hz, 1H), 7.86 (s, 1H), 7.82-7.78 (m, 1H), 7.72-7.61 (m, 2H), 7.58-7.55 (m, 1H), 7.42 (t, J = 7.9 Hz, 1H), 7.36 (t, J = 7.8 Hz, 2H), 7.14-7.11 (m, 1H), 6.91-6.88 (m, 1H), 6.61-6. 57(m,1H),4.02-3.99(m,1H),3.97–3.90(m,2H),3.80-3.76(m,1H),2.88(s,6H),2.32(d,J=11.8Hz,1H),2. 01(t,J=11.8Hz,2H),1.87(m,J=13.7,3.3Hz,1H),1.53–1.40(m,1H),1.41–1.28(m,2H),1.27-1.23(m,1H). 13 C NMR(151MHz,Methanol-d4)δ177.23,167.79,163.12,159.02,156.36,155.55,152.69,138.46,138.34,135.65,132.21,131.8 3,128.71,128.21,123.77,122.90,122.75,122.06,121.85,119.01,57.55,56.47,49.67,48.29,42.07,38.32,31.57,23.01.

[0056] Example 18: Preparation of N-(4-((5-chloro-4-(phenylamino)pyrimidin-2-yl)amino)cyclohexyl)-3-(cyclopropanecarboxamido)benzamide (4)

[0057] The operation was the same as in Example 1 to obtain 119 mg of white solid powder. 1 H NMR(600MHz,DMSO-d6)δ10.33(s,1H),8.66(s,1H),8.31–8.15(m,1H),7.97(s,2H),7.80( t,J=7.8Hz,3H),7.50(d,J=7.8Hz,1H),7.45-7.35(m,3H),7.07(t,J=7.4Hz,1H),7.02(s, 1H),3.77(s,1H),1.98(d,J=12.2Hz,2H),1.88(d,J=12.1Hz,2H),1.80(t,J=7.1Hz,1H),1 .44(q,J=12.2Hz,2H),1.36(q,J=12.8,12.2Hz,2H),1.28-1.25(m,1H),0.92–0.69(m,4H). 13 C NMR (151MHz, DMSO-d6) δ172.24, 166.08, 159.67, 156.01, 154.92, 139.75, 139.56, 136.08, 128.92, 128.63, 123.54, 122.30, 122.16, 121.98, 121.93, 118.81, 101.55, 54.07, 50.38, 49.57, 48.43, 36.24, 31.60, 23.23, 14.98, 7.68. Experimental Example 1 Cell proliferation inhibition activity test

[0058] MTT assay to detect the 48h IC of drugs on breast cancer cells 50 Value: The experiment set up solvent control group and drug experimental group. Each drug experimental group set up 8 concentrations, and each concentration set up 3 parallel wells. Each experiment was repeated three times. In the 96-well plate, the cell concentration added to each well was 1×10 5 100 μL of cell suspension containing 3×10 cells / mL, i.e., each well contains 3×10 3When inoculating, pay attention to make the cells evenly distributed in each well. To prevent liquid evaporation, cells are not inoculated in a circle of wells around the 96-well plate, and PBS is added to keep it moist. After the cells adhere, 100 μL of the target compound of different concentrations is added to each well in the drug experimental group. The 96-well plate was placed in an incubator at 37°C and 5% CO2 for continued culture, and the culture was terminated after 72 hours. After the cells were treated with drugs for 48 hours, the 96-well plate was taken out, and the MTT solution was added to each well. After continuing to incubate in the cell culture incubator for 4 hours, the absorbance value (A) of each well was measured at 570nm on an automatic microplate reader. The inhibition rate was calculated by the formula. A linear regression analysis was performed on the inhibition rate against the drug concentration, and the IC was calculated using the linear equation. 50 The test results were expressed as mean ± standard deviation, and the experimental results were statistically analyzed using SPSS15.0 software, with P < 0.05 indicating a significant difference. The results are shown in Table 1.

[0059] Table 1 Effects of the tested compounds at different concentrations on the proliferation of two breast cancer tumor cells

[0060]

Claims

1. A compound represented by formula I or a pharmaceutically acceptable salt thereof: in R1 is halogen; R2 is selected from hydrogen, cycloalkyl, alkyl, and the alkyl may be substituted by alkyl.

2. The compound according to claim 1 or its isomer or pharmaceutically acceptable salt, wherein R1 is selected from chlorine or bromine.

3. The compound according to claim 1 or its isomers, pharmaceutically acceptable salts thereof are selected from hydrogen, cyclopropyl, N,N-dimethylamino.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the following compounds:

5. A pharmaceutical composition comprising the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

6. Use of the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 5 in the preparation of a medicament for treating a CDK12-related disease.

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