FAK inhibitors and their combination drugs

Abstract

To provide a FAK inhibitor in the field of tumor treatment with improved pharmacokinetics and reduced metabolic toxicity of the drug.SOLUTION: The present invention provides a compound represented by formula (I-G) or an optical isomer, a tautomer, a pharmaceutically acceptable salt, a hydrate, or a solvate thereof, where, R1 and R5 are selected from hydrogen or a methoxy group, one or more of R1, R5, R20, and R21 are substituted with hydrogen or deuterium, R17, R18, and R19 are substituted with deuterium, and / or R22, R23, and R24 are substituted with deuterium.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] This invention belongs to the field of drug chemistry and specifically relates to FAK inhibitors and their concomitant drugs. [Background technology]

[0002] Focal adhesion kinase (FAK) is an intracellular non-receptor tyrosine kinase that was first discovered in transfected V-Src chicken embryo fibroblasts. FAK is highly expressed in many tissues, and its protein sequence exhibits high homology across many species (mice, toads, humans, etc.). FAK is a crossroads of multiple intracellular signaling pathways involved in several biological processes, including tumor formation, growth, metastasis and apoptosis, and cardiovascular disease, and is currently one of the most widely studied antitumor targets.

[0003] Recent research has revealed that FAK is activated by various factors, including integrins and G protein-coupled receptors, and that FAK regulates intracellular signaling pathways such as P53 and PI3K-AKT-mTOR via two kinase-dependent and kinase-independent pathways, thereby participating in biological processes such as tumor cell survival, proliferation, and metastasis. The initial attempt aimed to suppress tumors by downregulating FAK expression in tumor cells. Transfecting with carboxyl-termined FAK (FAK-CD) silences FAK, reducing cell adhesion and proliferation, and inhibiting breast cancer cell proliferation in in vivo experiments. Furthermore, transfecting plasmids containing FAK-silenced RNA (FAK-siRNA) suppresses cancer in vivo. Simultaneously, suppressing the expression of FAK and FAK downstream signaling molecules (e.g., SRC) can enhance the antitumor effect.

[0004] Due to the important function of FAK in tumor cells, the reliability of gene transfection, and the safety of viral vectors, small molecule inhibitors based on the FAK signaling pathway have emerged and have shown promising results in recent years. Currently, several FAK inhibitors are used as antitumor drugs and are in the preclinical or clinical trial stages. In the literature ("Research progress on FAK, a novel target adhesion plaque kinase for antitumor, and its inhibitors," Chen Ying et al.), it has been reported that TAE226, also known as NVP-226, can suppress FAK activity by blocking the binding site between FAK and ATP, and the phosphorylation sites of Y397 and Y861 of FAK. However, there is still a need in this field to develop FAK inhibitors with superior inhibitory activity or superior pharmacodynamic properties.

[0005] Deuterated drugs are drugs in which some of the hydrogen atoms in the drug molecule are replaced with deuterium. Deuterium (D) is a stable isotope of hydrogen, and its form and volume in drugs are essentially the same as hydrogen. Therefore, replacing some of the hydrogen atoms in a drug molecule with deuterium does not fundamentally change the activity of the drug molecule. Also, since the mass of a deuterium atom is twice that of hydrogen, the zero-point vibrational energy of the carbon-deuterium bond (CD) is lower than that of the carbon-hydrogen bond (CH), making the carbon-deuterium bond more stable. Therefore, by replacing some of the hydrogen atoms in a drug molecule with deuterium, the degradation process of the drug is slowed, allowing the deuterated drug to act longer in the body, and the objective of altering the drug metabolism rate or metabolic pathway is achieved, thereby improving pharmacokinetics and reducing the metabolic toxicity of the drug. In light of the important applications and limitations of FAK inhibitors in the field of oncology, it is possible to discover new molecular entities by combining them with deuterated drug technology, reducing the impact on liver and kidney function, and improving drug safety. Improving efficacy is a research trend that will drive further development of this type of drug, and it has great potential for application.

[0006] While drug combination therapy is an effective way to improve treatment outcomes, there are currently no reports of using deuterated FAK inhibitors in combination with other anticancer drugs or methods.

Summary of the Invention

[0007] To solve the above problems, the present invention provides a deuterated compound and its use as a FAK inhibitor, and further provides a concept of using the aforementioned deuterated compound in combination with other anticancer drugs.

[0008] The present invention provides a compound represented by formula (I) or an optical isomer, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof, JPEG0007861083000001.jpg4385

[0009] Here, the S ring is selected from an aromatic ring or a five-membered heterocyclic ring, A, B, X, Y, Z are each independently selected from carbon or nitrogen, E is absent or a methylene group, and / or, R6 is selected from hydrogen or absent, R7 is selected from hydrogen, nitrogen or absent, R8 is selected from a haloalkyl group or a halogen, or, R7 and R8 are bonded to form a ring, and / or, R9 is selected from -NMeSO2Me, -CONHOMe, -CONHMe, amide, hydrogen or absent, R 10 is selected from hydrogen or absent, R 11 is selected from -NHSO2Me, halogen, substituted piperazine or hydrogen, the substituent on the piperazine is an ethanol group, R 12 is selected from -SO2Me or hydrogen, R 13 is selected from -CONHMe, -CONHOMe, N-alkylsulfonamide, hydrogen or absent, or, R 11 and R 13 are bonded to form a ring, Dx shown in formula (I) represents that hydrogen on at least one carbon atom of the compound in parentheses is substituted with deuterium, and x is an integer of 1 or more.

[0010] Preferably, the compound has a structure shown by formula (I-A), TIFF0007861083000002.tif4494

[0011] Here, A, B, X, Y, and Z are each independently selected from carbon or nitrogen, E is absent or a methylene group, R1 and R5 are selected from hydrogen or a methoxy group, R2 and R4 are selected from hydrogen or a methoxy group selected from, R3 is selected from hydrogen, -CONHMe, an alkoxy amide, morpholine, a methoxy group, ethylamine or a sulfonamide, or R2 and R3 are bonded to form a ring, or R3 and R4 are bonded to form a ring, and / or, R6 is selected from hydrogen or absent, R7 is selected from hydrogen, nitrogen or absent, R8 is selected from a haloalkyl group or a halogen, or R7 and R8 are bonded to form a ring, and / or, R9 is selected from -NMeSO2Me, -CONHOMe, -CONHMe, an amide hydrogen or absent, R 10 is selected from hydrogen or absent, R 11 is selected from -NHSO2Me, a halogen, a substituted piperazine or hydrogen, the substituent on the piperazine is an ethanol group, R 12 is selected from -SO2Me or hydrogen, R 13 is selected from -CONHMe, an N-alkyl sulfonamide, hydrogen or absent, or, R 11 and R 13 are bonded to form a ring, or, R 13 and R3 and R4 are bonded to form a ring, Dx shown in formula (I-A) represents that the hydrogen on at least one carbon atom of the compound in parentheses is substituted with deuterium, and x is an integer of 1 or more.

[0012] Preferably, the compound has a structure shown in formula (I-B), TIFF0007861083000003.tif3989

[0013] Here, A, X, and Y are selected from carbon or nitrogen, E is absent, R 14 、R 15 、R16 These are hydrogen and C, which are independent of each other. 1-6 Alkyl alkyl group, C 3-6 Selected from cycloalkyl groups, methyl groups, ethyl groups, or isopropyl groups, and / or, R6 is hydrogen, R7 is hydrogen, R8 is halogen, R9, R 13 R is selected from -CONHOMe, -CONHMe, or hydrogen. 10 , R 12 It is hydrogen, R 11 is selected from halogen, hydrogen, or substituted piperazines, and the substituent on the piperazine is an ethanol group. In formula (IB), Dx represents that at least one hydrogen atom on the compound in parentheses is substituted with deuterium, and x is an integer greater than or equal to 1.

[0014] Preferably, the compound has the structure shown in formula (IC), TIFF0007861083000004.tif4897

[0015] Here, A, X, and Y are selected from carbon or nitrogen, and E is selected from a methylene group or none. Selected, R9, R 13 R is selected from hydrogen, -NMeSO2Me, -CONHOMe, or -CONHMe, 10 , R 12 is hydrogen, R 11 The substituent is selected from hydrogen, a substituted piperazine, or a halogen, and the substituent on the piperazine is an ethanol group. and / or R6 is selected from hydrogen, R7 is selected from hydrogen, R8 is selected from a haloalkyl group or halogen, or R7 and R8 are bonded together to form a ring. and / or R1, R4, R5 are selected from hydrogen or a methoxy group, R2 is selected from hydrogen or a methoxy group, R3 is selected from hydrogen, -CONHMe, alkoxyamide, morpholine, methoxy group, ethylamine or sulfonamide, or R2 and R3 are bonded to form a ring. In formula (IC), Dx represents that at least one hydrogen atom on the compound in parentheses is substituted with deuterium, and x is an integer greater than or equal to 1.

[0016] Preferably, the compound has the structure shown in formula (ID), TIFF0007861083000005.tif4386

[0017] Here, R9, R 13 R is selected from -CONHOMe, -CONHMe, or hydrogen. 10 , R 12 is hydrogen, R 11 The substituent is selected from halogen, hydrogen, or substituted piperazines, and the substituent on the piperazine is an ethanol group. A, X, and Y are selected from carbon or nitrogen, and E is none. and / or R6 is selected from hydrogen, R7 is selected from hydrogen, and R8 is halogen. and / or, R 14 R is selected from a methyl group, an ethyl group, or an isopropyl group. 15 R is selected from a methyl group or a hydrogen atom. 16 It is hydrogen, In formula (ID), Dx represents that at least one hydrogen atom on the compound in parentheses is substituted with deuterium, and x is an integer greater than or equal to 1.

[0018] Preferably, the compound has the structure shown in formula (IE), TIFF0007861083000006.tif4791

[0019] Here, B and Z are selected from carbon or nitrogen, E is a methylene group, Y is nitrogen, and X and A are carbon. and / or, R6 is none, R7 is hydrogen, and R8 is a haloalkyl group. and / or, R1, R2 are hydrogen, R3 is -CONHMe, R4 is hydrogen, or R3 and R4 are bonded to form a ring. In formula (IE), Dx represents that at least one hydrogen atom on the compound in parentheses is substituted with deuterium, and x is an integer greater than or equal to 1.

[0020] Preferably, the compound has the structure shown in formula (IF), TIFF0007861083000007.tif46102

[0021] Here, Dx in formula (IF) represents that at least one hydrogen atom on the compound in parentheses is substituted with deuterium, and x is an integer greater than or equal to 1.

[0022] Preferably, the compound has the structure shown in formula (IG), TIFF0007861083000008.tif3584

[0023] Here, R1, R5, R 17 , R 18 , R 19 , R 20 , R 21 , R 22 , R 23 , R 24 One or more of these are substituted with deuterium.

[0024] Preferably, the compound is selected from, but is not limited to, one of the following compounds substituted with deuterium. JPEG0007861083000009.jpg54153JPEG0007861083000010.jpg54159 TIFF0007861083000011.tif60162

[0025] Preferably, the compound is selected from, but is not limited to, one of the following compounds or one of the following compounds substituted with deuterium. TIFF0007861083000012.tif53164TIFF0007861083000013.tif51160 TIFF0007861083000014.tif24135 TIFF0007861083000015.tif78159TIFF0007861083000016.tif79163

[0026] The present invention also provides the use of the aforementioned compounds or their optical isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, or solvates in the preparation of FAK inhibitors, preferably FAK inhibitors which are drugs for the treatment of cancer.

[0027] Here, the cancer is a solid tumor, The solid tumors include mesothelioma, pancreatic cancer, soft tissue tumors, metastatic tumors, non-solid tumors, sarcomas, adenocarcinomas, lung cancer, breast cancer, lymphoma, gastrointestinal cancer, genitourinary cancer, prostate cancer, and ovarian cancer; the gastrointestinal cancers include colon cancer; the genitourinary cancers include kidney, urothelial, or testicular tumors; and the ovarian cancers include advanced ovarian cancer. The aforementioned mesotheliomas include neurofibromas, kidney cancers, lung cancers, small cell lung cancers, non-small cell lung cancers, KRAS-mutated non-small cell lung cancers, liver cancers, thyroid cancers, breast cancers, nervous system tumors, schwannomas, meningiomas, neuromas, adenoid cystic carcinomas, ependymomas, ependymal tumors, malignant pleural tumors, malignant pleural mesotheliomas, triploidomas, negative breast cancers, non-hematological malignancies, melanomas, colorectal cancers, leukemias, adenocarcinomas, and solid tumors. The term "melanoma" includes locally advanced melanoma, locally mutated N-Ras-induced melanoma, and metastatic malignant cutaneous melanoma; the term "colorectal cancer" includes metastatic colorectal cancer; the term "leukemia" includes acute myeloid leukemia; the term "adenocarcinoma" includes adenocarcinoma; and the term "solid tumor" includes locally advanced solid tumor, metastatic solid tumor, and hepatocellular carcinoma.

[0028] The present invention also provides combination drugs for treating tumors, comprising the aforementioned compounds and anticancer drugs used for simultaneous or separate administration in the same or different standard unit formulations, as well as pharmaceutically acceptable carriers.

[0029] The aforementioned anticancer drug is an immunotherapy drug, a chemotherapy drug, or a radiotherapy drug.

[0030] The immunotherapy drug is selected from checkpoint inhibitors, PD-1 inhibitors, PD-L1 inhibitors, antibodies that inhibit CTLA-4, antibodies that inhibit TIM3, antibodies that inhibit LAG3, antibodies that inhibit TIGIT, checkpoint-targeted blockade antibodies, costimulatory antibodies, or CAR-T therapy cells.

[0031] The PD-1 inhibitor or PD-L1 inhibitors include nivolumab, CT-011, AMP-224, pembrolizumab, pidilizumab, MK-3475, BMS936559, MEDI4736, and MSB001071. This includes, but is not limited to, 8C, MPDL-3280A, SHR-1210, IBI308, BGB-A317, JS001, GLS-010, GB226 geptanolimab, HLX10, AK103, AK104, AK105, AK112, SSI-361, JY034, KN035, SHR1316, TQB2450, KL-A167, CS1001, STI-A1014, JS003, AK106, HLX-09, and mPD-1 antibodies. The aforementioned checkpoint-targeted blocking antibodies include IMP321 and MGA271. The aforementioned costimulatory antibodies include anti-4-IBB antibody, anti-OX40 antibody, anti-GITR antibody, anti-CD27 antibody, and anti-CD40 antibody.

[0032] The aforementioned chemotherapy drugs include toxic agents, alkylating agents, antimetabolites, antibiotics, hormonal therapy agents, natural product anticancer agents, topoisomerase inhibitors, immunotherapy agents, platinum complex agents, kinase inhibitors, antiproliferative agents, antibodies, interferons, or drugs that modulate androgen signaling pathways.

[0033] The aforementioned toxic agents include, but are not limited to, gemcitabine, paclitaxel, and docetaxel. The kinase inhibitors mentioned above include, but are not limited to, MEK kinase inhibitors, cMet inhibitors, VEGFR2 inhibitors, and EGFR inhibitors. Drugs that modulate the androgen signaling pathway include, but are not limited to, androgen synthesis inhibitors, CYP17A inhibitors, androgen receptor inhibitors, BET inhibitors, BRD4 inhibitors, RORγ inhibitors, CBP / P300 inhibitors, BMX inhibitors, and PARP inhibitors. Preferably, the androgen receptor inhibitors include, but are not limited to, Enzalutamide, Apalutamide, Bicalutamide, Abiraterone, ODM-201, EPI-001, ONC1-13B, EM-5854, JNJ-63576, TAS-3681, HC-1119, Proxyalutamide, and SHR3680.

[0034] The present invention also provides the use of the aforementioned combination drugs in the preparation of drugs for treating cancer. do.

[0035] Here, the cancer is a solid tumor, The solid tumors include mesothelioma, pancreatic cancer, soft tissue tumors, metastatic tumors, non-solid tumors, sarcomas, adenocarcinomas, lung cancer, breast cancer, lymphoma, gastrointestinal cancer, genitourinary cancer, prostate cancer, and ovarian cancer; the gastrointestinal cancers include colon cancer; the genitourinary cancers include kidney, urothelial, or testicular tumors; and the ovarian cancers include advanced ovarian cancer. The aforementioned mesotheliomas include neurofibromas, kidney cancers, lung cancers, small cell lung cancers, non-small cell lung cancers, KRAS-mutated non-small cell lung cancers, liver cancers, thyroid cancers, breast cancers, nervous system tumors, schwannomas, meningiomas, neuromas, adenoid cystic carcinomas, ependymomas, ependymal tumors, malignant pleural tumors, malignant pleural mesotheliomas, triploidomas, negative breast cancers, non-hematological malignancies, melanomas, colorectal cancers, leukemias, adenocarcinomas, and solid tumors. The term "melanoma" includes locally advanced melanoma, locally mutated N-Ras-induced melanoma, and metastatic malignant cutaneous melanoma; the term "colorectal cancer" includes metastatic colorectal cancer; the term "leukemia" includes acute myeloid leukemia; the term "adenocarcinoma" includes adenocarcinoma; and the term "solid tumor" includes locally advanced solid tumor, metastatic solid tumor, and hepatocellular carcinoma.

[0036] In the present invention, "alkyl group" includes linear or branched alkyl groups.

[0037] In the present invention, the term "compound of the present invention" refers to the compound shown in formula (I). This term also includes various crystalline forms of the compound of formula (I), pharmaceutically acceptable salts, hydrates or solvates, optical isomers, tautomers, and prodrugs.

[0038] In this invention, the term "pharmaceutically acceptable salt" means a salt suitable for pharmaceutical use, formed with the compound of the present invention and an acid or base. Pharmaceutically acceptable salts include inorganic and organic salts. Preferred salts are those formed with the compound of the present invention and an alkali metal. Suitable alkali metals for salt formation include, but are not limited to, lithium, sodium, potassium, calcium, and magnesium.

[0039] The method of administering the compound or drug composition of the present invention is not particularly limited, and typical methods of administration include (but are not limited to) oral, parenteral (intravenous, intramuscular, or subcutaneous), and topical administration.

[0040] The present invention provides a deuterated compound that exhibits superior pharmacokinetics, higher peak blood concentrations, higher exposure levels, and longer half-lives compared to the compound before deuteration, and thus possesses superior metabolic performance. Furthermore, the deuterated compound of the present invention can effectively inhibit FAK activity, making it highly promising for application in the preparation of FAK inhibitors and / or drugs for treating cancer. At the same time, the deuterated compound of the present invention can exert a synergistic effect when used in combination with anticancer drugs (e.g., PD-1 inhibitors), significantly improving tumor suppression effects and providing a superior option in clinical cancer treatment.

[0041] Naturally, based on the above-described content of the present invention, and in light of general technical knowledge and conventional means in the art, various other forms of modifications, substitutions, or changes can be made, provided that they do not deviate from the above-described basic technical idea of ​​the present invention.

[0042] The above-mentioned content of the present invention will be described in more detail below by means of embodiments for carrying out the invention. However, this should not be understood as limiting the scope of the subject matter of the present invention to the following embodiments. Any technology realized based on the above-mentioned content of the present invention falls within the scope of the present invention. [Brief explanation of the drawing]

[0043] [Figure 1] Figure 1 shows the efficacy test of the deuterated compound of the present invention in an animal model of MC38 tumor. [Figure 2] Figure 2 shows the efficacy testing of the deuterated compound of the present invention in an animal model of PAN02 tumor. [Modes for carrying out the invention]

[0044] All raw materials and equipment used in this invention are commercially available products.

[0045] Example 1: Synthesis of N-triduteromethyl-4-((4-(((3-(N-methylmethanesulfonamide)pyrazine-2-yl)methyl)amino)-5-(trifluoromethyl)pyrimidine-2-yl)amino)benzoic acid amide (Compound 25) TIFF0007861083000017.tif55169

[0046] Step 1: Synthesis of Compound 25-2 25-1 (200 mg, 0.39 mmol) and DMAP (1.29 g, 10.57 mmol) were added to 10 mL of dichloromethane, and then (Boc)2O (1.71 g, 7.83 mmol) was added dropwise. The system was refluxed in an oil bath for 24 hours. The next day, it was cooled to room temperature, dichloromethane and 0.1 N HCl solution were added, and the mixture was extracted. After standing, the layers were separated, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered by suction, and the solvent was removed by rotary evaporation. The crude product was separated by column chromatography to obtain a white solid 25-2 (136 mg, yield: 42.8%). MS(M+1): 811.2.

[0047] Step 2: Synthesis of Compound 25-3 25-2 (136 mg, 0.17 mmol) and deuterated methylamine hydrochloride (189 mg, 2.68 mmol) were added to 5 mL of acetonitrile and stirred at room temperature. Then, DBU (613 mg, 4.03 mmol) was added and gradually dissolved to clarify the mixture. Next, the system was placed in an oil bath and refluxed overnight. The following day, it was cooled to room temperature, the solvent was removed by rotary evaporation, and then dichloromethane and 0.1 N HCl solution were added to the system. The mixture was stirred vigorously, allowed to stand to separate the layers, and the organic phase was washed with purified water and saturated brine, respectively. The mixture was dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the mixture was separated and purified by pre-TLC (PE / EA = 2:1) to obtain a white solid 25-3 (42 mg, yield 35.3%). MS(M-Boc+1): 614.2.

[0048] Step 3: Synthesis of Compound 25 25-3 (42 mg, 0.06 mmol) was added to 2 mL of dichloromethane and stirred at room temperature (it did not dissolve and was not clarified). Next, 0.1 mL of trifluoromethanesulfonic acid was added, and the system was gradually clarified and reacted by stirring at room temperature overnight. The next day, the solvent was removed by rotary evaporation, and ethyl acetate and saturated NaHCO3 solution were added to the system, stirred vigorously, and then allowed to stand to separate the layers. The measured pH value of the aqueous phase was approximately 7-8. The organic layer was washed twice each with water and saturated brine, and dried over anhydrous sodium sulfate. The solvent was removed by rotary evaporation, yielding compound 25 (24 mg, yield: 80.0%) as a white solid.

[0049] 1 H NMR(400Hz,DMSO-d6):δ9.863(1H,s),8.688(1H,d,J=2.4Hz),8.581(1H,d,J=2.8Hz),δ8.316(1H,s),δ8.180(1H,s),7.665-7.59 4(4H,dd,J1=19.6Hz,J2=8.8Hz),7.476-7.450(1H,t,J=5.2Hz),δ5.001(2H,d,J=4.8Hz),3.221(3H,s),3.199(3H,s).LC-MS(M+H + ): 514.2.

[0050] Compounds 26-40 were produced using raw materials corresponding to each compound and a method similar to the method used to produce compound 25.

[0051] Example 2 Synthesis of N-methyl-4-((4-(((3-(N-trideuteromethylmethanesulfonamide)pyrazine-2-yl)methyl)amino)-5-(trifluoromethyl)pyrimidine-2-yl)amino)benzoic acid amide (compound 41) TIFF0007861083000018.tif60154

[0052] Step 1: Synthesis of N-trideuteromethylmethanesulfonamide (compound 41-1) 7.75 g, 109.99 mmol of deuterated methylamine hydrochloride was placed in a 250 mL round-bottom flask, and 120 mL of dichloromethane was added simultaneously. The mixture was stirred at room temperature. The system was then transferred to an ice bath for cooling and stirred while lowering the temperature. After 15 minutes, triethylamine (21.73 g, 214.75 mmol) and DMAP (128 mg, 1.05 mmol) were added in sequence. After completion, the system was kept in the ice bath, stirring to maintain the temperature, and allowed to react for 10 minutes. Then, methanesulfonyl chloride (12.0 g, 104.76 mmol) was added to the system. After completion, the ice bath was removed, and the system was stirred at room temperature overnight. The next day, the completion of the reaction was detected by TLC. The system was then subjected to suction filtration, and small amounts of ethyl acetate were used to elute the filtrate multiple times. The filtrates were combined, the solvent was removed by rotary evaporation, and 90 mL of ethyl acetate was added to the system and vigorously stirred. After 10 minutes, the system was again subjected to suction filtration. The filtrate cake was eluted multiple times with small amounts of ethyl acetate, and the filtrates were combined and concentrated under vacuum to obtain a colorless, transparent oily liquid of N-trideuteromethylmethanesulfonamide (11.16 g). No further purification was performed, and this was used directly in the next reaction. Yield: 94.9%. LC / MS (ESI+)Calcd for C2H4D3NO2S(M+H+)m / z, 113.1;Found: 113.3.

[0053] Step 2: Synthesis of N-(3-cyanopyrazine-2-yl)-N-trideuteromethylmethanesulfonamide (compound 41-2) N-trideuteromethylmethanesulfonamide (6.0g, 53.54mmol), 2-chloro-3-cyanopyrazine (6.23 g, 44.62 mmol) was placed in a 500 mL round-bottom flask, and acetonitrile (300 mL) was added simultaneously. The mixture was stirred at room temperature. Next, cesium carbonate (24.71 g, 75.85 mmol) was added to the system. After completion, the system was transferred to an 80°C oil bath and the reaction was continued with heating and stirring. After 1.5 hours, the sample was dropped onto a plate, and the TLC showed that the raw materials had been completely consumed. Heating was stopped, and the system was allowed to cool naturally to room temperature. The system was subjected to suction filtration, and the filtrate cake was eluted multiple times using small amounts of acetonitrile. The filtrates were then combined, and the solvent was removed by rotary evaporation. Then, ethyl acetate (150 mL) and water (150 mL) were added to the system, and the mixture was vigorously stirred. After standing, the layers were separated, and the aqueous phase was back-extracted with ethyl acetate (50 mL x 3). The organic phase was combined with purified water (30 mL x 3) and saturated brine (30 mL), and the mixture was washed sequentially with purified water (30 mL x 3) and saturated brine (30 mL). The mixture was dried over anhydrous sodium sulfate and concentrated under vacuum to obtain the crude product. This was separated and purified by column chromatography to obtain a pale reddish-brown oily liquid of N-(3-cyanopyrazine-2-yl)-N-deuteromethylmethanesulfonamide (5.29 g). Yield: 55.1%. LC / MS (ESI+) Calcd for C7H5D3N4O2S(M+H+)m / z, 216.1; Found: 233.1(M+H2O). 1H NMR (400MHz, DMSO-d6) δ8.65-8.63 (dd, J=6.0, 2.4Hz, 2H), 3.26 (s, 3H).

[0054] Step 3: Synthesis of N-(3-(aminomethyl)pyrazine-2-yl)-N-trideuteromethylmethanesulfonamide (compound 41-3) N-(3-cyanopyrazine-2-yl)-N-deuteromethylmethanesulfonamide (2.0 g, 9.30 mmol) was weighed into a 500 mL round-bottom flask, methanol (270 mL) was added, and the mixture was stirred at room temperature to dissolve and clarify. Next, wet palladium carbon (1 g) and aqueous ammonia (20 mL) were added to the system. Then, the system was evacuated and argon gas was introduced five times to ensure an inert gas atmosphere. The system was then subjected to hydrogenation, and the reaction was allowed to continue with stirring at room temperature after completion. After 5 hours, the sample was dropped onto a plate, and the TLC showed that the raw materials had been completely consumed. The reaction was stopped, and the hydrogenation apparatus was removed. The system was subjected to suction filtration, and the filtrate cake was repeatedly eluted with methanol. The filtrates were combined, the solvent was removed by rotary evaporation, and residual water in the system was removed multiple times using a rotary band with methanol to obtain a pale yellowish-brown, transparent oily liquid of N-(3-(aminomethyl)pyrazine-2-yl)-N-deuteromethylmethanesulfonamide. No further purification was performed, and this was used directly in the next reaction. LC / MS (ESI+)Calcd for C7H9D3N4O2S(M+H+)m / z,220.1;Found:220.1.

[0055] Step 4: Synthesis of the compound tert-butyl(4-(methylcarbamoyl)phenyl)carbamate 4-((tert-butoxycarbonyl)amino)benzoic acid (3.0 g, 12.65 mmol) was weighed into a 250 mL round-bottom flask, and 50 mL of DMF was added. The mixture was stirred at room temperature. Then, EDCI (4.8 g, 25.29 mmol), TEA (4.5 g, 44.28 mmol), methylamine hydrochloride (1.3 g, 18.98 mmol), and DMAP (16.0 mg, 0.13 mmol) were added to the system in order. After completion, the system was stirred at room temperature overnight to allow the reaction to proceed. The following day, monitoring confirmed that all raw materials had been consumed. Ethyl acetate (70 mL) and water (50 mL) were added to the system, vigorously stirred, and allowed to stand to separate the layers. The aqueous phase was back-extracted with ethyl acetate (20 mL x 3), the organic layers were combined, washed with water (20 mL x 3) and saturated brine (30 mL) respectively, dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation to obtain the crude product. Subsequently, it was separated by column chromatography to obtain 2.1 g of off-white solid tert-butyl(4-(methylcarbamoyl)phenyl)carbamate. Yield: 66.5%. MS(ESI)m / e25 1.2(M+H) + .

[0056] Step 5: Synthesis of compound 4-amino-N-methylbenzamidetrifluoroacetate 500.0 mg of tert-butyl(4-(methylcarbamoyl)phenyl)carbamate (2.00 mmol) was weighed into a 50 mL round-bottom flask, and 10 mL of dichloromethane was added. The mixture was stirred at room temperature. Then, 1 mL of trifluoroacetic acid was added to the system. After completion, the system was stirred at room temperature overnight to allow the reaction to proceed. The next day, TLC showed that the reaction was complete. The mixture was concentrated to remove the solvent and excess trifluoroacetic acid, and any remaining trifluoroacetic acid in the system was removed multiple times using a rotating band with dichloromethane until the system was completely solid, yielding a white solid 4-amino-N-methylbenzamidetrifluoroacetate (510.0 mg). No further purification was performed, and this was used directly in the next reaction.

[0057] Step 6: Synthesis of compound 4-((4-chloro-5-(trifluoromethyl)pyrimidine-2-yl)amino)-N-methylbenzamide 499.0 mg of 2,4-dichloro-5-(trifluoromethyl)pyrimidine (2.30 mmol) was weighed into a 50 mL round-bottom flask, and 5 mL of 1,2-dichloroethane and 5 mL of tert-butanol were added to the system. The mixture was stirred at room temperature to dissolve and clarify. The system was then transferred to an ice bath and stirred continuously while cooling and lowering the temperature. After 15 minutes, zinc bromide (1.4 g, 6.00 mmol) was added to the system. After completion, the system was stirred in an ice bath for 30 minutes while maintaining the temperature. Subsequently, 4-amino-N-methylbenzamide trifluoroacetate and triethylamine (648.0 mg, 6.40 mmol) synthesized in the previous step were added to the system. After adding, the ice bath was removed, and the system was stirred at room temperature overnight to allow the reaction to proceed. The following day, monitoring confirmed the completion of the reaction. The solvent was removed by rotary evaporation, ethyl acetate (30 mL) and water (20 mL) were added to the system, and after vigorous stirring and standing to separate the layers, the aqueous layer was back-extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed sequentially with water (15 mL x 3) and saturated brine (15 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. This was then separated by column chromatography to obtain 280.0 mg of a white solid 4-((4-chloro-5-(trifluoromethyl)pyrimidine-2-yl)amino)-N-methylbenzamide. Yield: 42.3%. MS(ESI)m / e331.0(M+H) + .

[0058] Step 7: Synthesis of compound N-methyl-4-((4-(((3-(N-deuteromethylmethanesulfonamide group)pyrazine-2-yl)methyl)amino)-5-(trifluoromethyl)pyrimidine-2-yl)amino)benzamide In a 25 mL round-bottom flask containing N-(3-(aminomethyl)pyrazine-2-yl)-N-deuteromethylmethanesulfonamide (compound 41-3, 65.8 mg, 0.30 mmol), 5 mL of 1,2-dichloroethane and 5 mL of tert-butanol were added and stirred at room temperature to dissolve, then clarified. Next, 4-((4-chloro-5-(trifluoromethyl)pyrimidine-2-yl)amino)-N-methylbenzamide (100.0 mg, 0.30 mmol) and diisopropylethylamine (116.3 mg, 0.90 mmol) were added sequentially to the system. After adding these, the system was transferred to an oil bath at 80°C and refluxed. After 8 hours, TLC monitoring confirmed that the raw materials had been completely consumed. Heating was stopped, and after the system had cooled to room temperature, the solvent was removed by rotary evaporation to obtain the crude product. Subsequently, this was separated and purified by pre-TLC to obtain 12 mg of the off-white solid N-methyl-4-((4-(((3-(N-triduteromethylmethanesulfonamide)pyrazine-2-yl)methyl)amino)-5-(trifluoromethyl)pyrimidine-2-yl)amino)benzamide (compound 41). Yield: 7.8%. MS(ESI)m / e514.2(M+H) + . 1 H NMR(400MHz,DMSO-d6)δ9.83(s,1H),8.69(s,1H),8.59(s,1H),8.32(s,1H),8.20(d ,J=4.0Hz,1H),7.68-7.61(dd,J=14.4,8.4Hz,4H),7.41-7.39(t,J=4.4Hz,1H),5.01(d,J=3.6Hz,2H),3.20(s,3H),2.76(d,J=4.0Hz,3H).

[0059] Compounds 42 and 43 were produced using raw materials corresponding to each compound and a method similar to the method used to produce compound 41.

[0060] Example 3 Synthesis of N-triduteromethyl-4-((4-(((3-(N-triduteromethylmethanesulfonamide)pyrazine-2-yl)methyl)amino)-5-(trifluoromethyl)pyrimidine-2-yl)amino)benzoic acid amide (compound 44) ​​and its hydrochloride salt TIFF0007861083000019.tif58159

[0061] Step 1: Synthesis of tert-butyl (4-(triduteromethylcarbamoyl)phenyl)carbamate (compound 44-1) N-Boc-4-aminobenzoic acid (6.0 g, 25.29 mmol) and EDCI (7.27 g, 37.93 mmol) were weighed into a 250 mL round-bottom flask, and DMF (50 mL) was added simultaneously. The mixture was stirred at room temperature. Next, triethylamine (6.40 g, 63.22 mmol) and deuterated methylamine hydrochloride (1.96 g, 27.82 mmol) were added to the system. After completion, the system was stirred at room temperature overnight to allow the reaction to proceed. The next day, the sample was dropped onto a plate, and TLC indicated that the reaction was complete. Ethyl acetate (50 mL) and water (50 mL) were added to the system, and the mixture was stirred vigorously. After standing, the layers were separated, the aqueous phase was back-extracted with ethyl acetate (50 mL x 3), the organic phase was combined, and the mixture was washed sequentially with water (30 mL x 3) and saturated brine (50 mL). The mixture was dried over anhydrous sodium sulfate and concentrated under vacuum to obtain the crude product. This was separated and purified by column chromatography to obtain a white solid, tert-butyl (4-(deuteromethylcarbamoyl)phenyl)carbamate (4.92 g). Yield: 76.8%. LC / MS (ESI+)Calcd for C13H15D3N2O3(M+H+)m / z, 254.2;Found: 254.2.

[0062] Step 2: Synthesis of 4-amino-N-trideuteromethylbenzamidetrifluoroacetate (compound 44-2) 3.0 g, 11.84 mmol of tert-butyl (4-(deuteromethylcarbamoyl)phenyl)carbamate was weighed into a 100 mL round-bottom flask, and 15 mL of dichloromethane was added simultaneously. The mixture was stirred at room temperature. Next, 7 mL of trifluoroacetic acid was added to the system. After completion, the system was stirred at room temperature to allow the reaction to proceed. After 5 hours, the sample was dropped onto a plate, and TLC indicated that the reaction was complete. The solvent and excess trifluoroacetic acid were removed by rotational evaporation, and the remaining trifluoroacetic acid was repeatedly removed using dichloromethane and a rotational band to obtain a white solid 4-amino-N-deuteromethylbenzamidetrifluoroacetate. No further purification was performed, and it was used directly in the next reaction. I used this.

[0063] Step 3: Synthesis of 4-((4-chloro-5-(trifluoromethyl)pyrimidine-2-yl)amino)-N-triduteromethylbenzamide (compound 44-3) 2,4-Dichloro-5-trifluoromethylpyrimidine (2.83 g, 13.02 mmol) was weighed into a 100 mL round-bottom flask, and 1,2-dichloroethane (30 mL) and tert-butanol (30 mL) were added simultaneously. The mixture was stirred at room temperature to dissolve and clarify. The system was then transferred to an ice bath and cooled, stirring continuously while lowering the temperature. After the temperature in the system had dropped to approximately 0°C, zinc bromide (8.0 g, 35.52 mmol) was added to the system. After completion, the system was allowed to react for 30 minutes while maintaining the temperature in an ice bath. Next, 4-amino-N-deuteromethylbenzamidetrifluoroacetate and triethylamine (3.83 g, 37.89 mmol) prepared in the previous step were added to the system. The ice bath was then removed, and the system was stirred at room temperature overnight to allow the reaction to proceed. The following day, when the sample was dropped onto a plate, the TLC indicated that the raw materials had been completely consumed, and the reaction was stopped. The solvent was removed by rotary evaporation, ethyl acetate (50 mL) and water (30 mL) were added to the system, and the mixture was vigorously stirred. After standing, the layers were separated, the aqueous phase was back-extracted with ethyl acetate (30 mL x 3), the organic phase was combined, and the mixture was washed sequentially with water (30 mL x 3) and saturated brine (30 mL). The mixture was dried over anhydrous sodium sulfate and concentrated in vacuum to obtain the crude product. This was separated and purified by column chromatography to obtain the off-white solid 4-((4-chloro-5-(trifluoromethyl)pyrimidine-2-yl)amino)-N-deuteromethylbenzamide (3.23 g). Yield of the reaction in steps 2-3: 81.8%.

[0064] LC / MS(ESI+)Calcd for C13H7D3ClF3N4O(M+H+)m / z,334.0;Found:334.0.1H NMR (400MHz, DMSO-d6) δ10.89(s, 1H), 8.87(s, 1H), 8.31(s, 1H), 7.84-7.77(m, 4H).

[0065] Step 4: Synthesis of N-triduteromethyl-4-((4-(((3-(N-triduteromethylmethanesulfonamide)pyrazine-2-yl)methyl)amino)-5-(trifluoromethyl)pyrimidine-2-yl)amino)benzamide (compound 44) 4-((4-chloro-5-(trifluoromethyl)pyrimidine-2-yl)amino)-N-deuteromethylbenzamide (3.1 g, 9.29 mmol) and N-(3-cyanopyrazine-2-yl)-N-deuteromethylmethanesulfonamide (2.0 g, 9.29 mmol) were weighed into a 250 mL round-bottom flask. Then, 1,2-dichloroethane (80 mL) and tert-butanol (80 mL) were added, and the mixture was stirred at room temperature to dissolve and clarify the solution. After that, diisopropylethylamine (3.6 g, 27.87 mmol) was added to the system, and once complete, the system was transferred to an oil bath at 80°C and incubated overnight under reflux and stirring. The next day, when the sample was added dropwise to a plate, TLC showed that the reaction was complete. The solvent was removed by rotary evaporation, ethyl acetate (100 mL) and water (50 mL) were added to the system, and the mixture was vigorously stirred. After standing, the layers were separated, the aqueous phase was back-extracted with ethyl acetate (50 mL x 3), the organic phase was combined, and the mixture was washed sequentially with water (30 mL x 3) and saturated brine (50 mL). The mixture was dried over anhydrous sodium sulfate and concentrated under vacuum to obtain the crude product. This was separated and purified by column chromatography to obtain the off-white solid target compound (2.36 g). This solid was then placed in a 250 mL round-bottom flask, ethyl acetate (75 mL) was added, and the mixture was stirred at room temperature to form a slurry. After 3 hours, the solution was subjected to suction filtration, and the filtration cake was eluted multiple times with small amounts of ethyl acetate (45 mL). The mixture was then dried in a vacuum drying oven at a low temperature to obtain a white solid N-duteromethyl-4-((4-(((3-(N-duteromethylmethanesulfonamide)pyrazine-2-yl)methyl)amino)-5-(trifluoromethyl)pyrimidine-2-yl)amino)benzamide (2.11 g). Yield: 44.0%. LC / MS (ESI+)Calcd for C20H15D6F3N8O3S(M+H+)m / z,517.2;Found:517.2.1H NMR(400MHz,DMSO)δ9.83(s,1H),8.69(d,J=2.8Hz,1H),8.58(d,J=2.4Hz,1H),8.31(s,1H),8.17(s ,1H),7.67-7.61(dd,J=15.4,8.6Hz,4H),7.41(t,J=5.0Hz,1H),5.00(d,J=4.8Hz,2H),3.20(s,3H).

[0066] Step 5: Synthesis of N-duteromethyl-4-((4-(((3-(N-duteromethylmethanesulfonamide)pyrazine-2-yl)methyl)amino)-5-(trifluoromethyl)pyrimidine-2-yl)amino)benzamide hydrochloride Compound 44 (500 mg, 0.97 mmol) was weighed into a 100 mL round-bottom flask, and methanol (25 mL) was added. The mixture was then heated to room temperature and mixed thoroughly. Next, a solution of hydrogen chloride in ethanol (2.25 mL, 2.0 M) was slowly added dropwise to the system. After completion, the system was continued to react with stirring at room temperature. After 1.5 hours, the system was subjected to suction filtration. The filtration cake was eluted multiple times using small amounts of methanol (15 mL), and the mixture was dried in a vacuum drying oven at a low temperature to obtain a white solid N-duteromethyl-4-((4-(((3-(N-duteromethylmethanesulfonamide)pyrazine-2-yl)methyl)amino)-5-(trifluoromethyl)pyrimidine-2-yl)amino)benzoyl hydrochloride (517 mg). Yield: 96.6%. LC / MS(ESI+)Calcd for C20H16D6ClF3N8O3S(M+H+)m / z,517.2;Found:517.2.1H NMR(400MHz,DMSO)δ10.02(s,1H),8.68(d,J=2.8Hz,1H),8.58(d,J=2.4Hz,1H),8.35(s ,1H),8.21(s,1H),7.67-7.57(m,5H),5.26(br,6H)5.00(d,J=4.8Hz,2H),3.19(s,3H).

[0067] Example 4 Synthesis of N-duteromethyl-4-((4-(((3-(N-duteromethylmethanesulfonamide)pyrazine-2-yl)duteromethyl)amino)-5-(trifluoromethyl)pyrimidine-2-yl)amino))benzamide (Compound 45) TIFF0007861083000020.tif45169

[0068] Step 1: Synthesis of compound N-(3-(aminodiduteromethyl)pyrazine-2-yl)-N-duteromethylmethanesulfonamide (45-1) Compound 41-2 (100.0 mg, 0.46 mmol) was weighed into a 25 mL round-bottom flask, and 5 mL of deuterated methanol was added. The mixture was stirred at room temperature to dissolve and clarify. Next, 20.0 mg of wet palladium carbon (treated with heavy water) and triethylamine (188.2 mg, 1.86 mmol) were added sequentially to the system, and the hydrogenation procedure was repeated 10 times. After completion, the system was stirred at room temperature to allow the reaction to proceed. After 72 hours, the completion of the reaction was confirmed by monitoring. The system was subjected to suction filtration, and deuterated methanol (10 mL) was eluted multiple times in small amounts from the filtrate cake. The filtrates were combined, and the solvent was removed by rotary evaporation to obtain a pale yellowish-brown oily liquid, N-(3-(aminoduteromethyl)pyrazine-2-yl)-N-duteromethylmethanesulfonamide. No further purification was performed, and this was used directly in the next reaction. MS(ESI)m / e222 .2(M+H) + .

[0069] Step 2: Synthesis of compound N-duteromethyl-4-((4-(((3-(N-duteromethylmethanesulfonamide)pyrazine-2-yl)duteromethyl)amino)-5-(trifluoromethyl)pyrimidine-2-yl)amino))benzamide (45) 2 mL of 1,2-dichloroethane and 2 mL of tert-butanol were added to a 25 mL round-bottom flask containing N-(3-(aminoduteromethyl)pyrazine-2-yl)-N-duteromethylmethanesulfonamide (22.1 mg, 0.10 mmol). The mixture was stirred at room temperature to dissolve and clarify. Compound 44-3 (33.4 mg, 0.10 mmol) and diisopropylethylamine (30.6 mg, 0.30 mmol) were then added to the system in sequence. After adding the compounds, the system was transferred to an oil bath at 80°C and refluxed. After 8 hours, TLC monitoring confirmed that the raw materials had been completely consumed. After stopping heating and allowing the system to cool to room temperature, the solvent was removed by rotary evaporation to obtain the crude product. This was then separated and purified by pre-TLC to obtain 8.1 mg of a white solid N-duteromethyl-4-((4-(((3-(N-duteromethylmethanesulfonamide)pyrazine-2-yl)methyl)amino)-5-(trifluoromethyl)pyrimidine-2-yl)amino))benzamide. Yield: 15.6%. MS(ESI)m / e519.2(M+H) + . 1 H NMR(400MHz,DMSO-d6)δ9.83(s,1H),8.69(d,J=2.0Hz,1H),8.58(d,J=2.4Hz,1H),8. 31(s,1H),8.17(s,1H),7.67-7.61(dd,J=15.2,8.8Hz,4H),7.39(s,1H),3.20(s,3H).

[0070] Example 5 Synthesis of 4-((4-(((3-(N-triduteromethylmethanesulfonamide)pyrazine-2-yl)methyl)amino)-5-(trifluoromethyl)pyrimidine-2-yl)amino)benzamide (Compound 52) TIFF0007861083000021.tif36170

[0071] Step 1: Synthesis of compound 4-((4-chloro-5-(trifluoromethyl)pyrimidine-2-yl)amino)benzamide(52-2) 434 mg of 2,4-dichloro-5-(trifluoromethyl)pyrimidine (2.00 mmol) was weighed into a 50 mL round-bottom flask, and 5 mL of 1,2-dichloroethane and tert-butanol were added to the system. The mixture was stirred at room temperature to dissolve and clarify. The system was then transferred to an ice bath and stirred continuously while cooling and lowering the temperature. After 15 minutes, zinc bromide (1.2 g, 5.22 mmol) was added to the system. After completion, the system was stirred in an ice bath for 30 minutes while maintaining the temperature. Subsequently, 237 mg of 4-aminobenzamide (237 mg, 1.74 mmol) and triethylamine (564 mg, 5.57 mmol) synthesized in the previous step were added to the system. After adding the compounds, the ice bath was removed, and the system was stirred overnight at room temperature to allow the reaction to proceed. The following day, monitoring confirmed the completion of the reaction. The solvent was removed by rotary evaporation, ethyl acetate (30 mL) and water (20 mL) were added to the system, and the mixture was vigorously stirred. After standing, the layers were separated, and the aqueous layer was back-extracted with ethyl acetate (10 mL x 3). The organic phases were combined and washed sequentially with water (15 mL x 3) and saturated brine (15 mL). The mixture was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. This was then separated by column chromatography to obtain 294 mg of the off-white solid 4-((4-chloro-5-(trifluoromethyl)pyrimidine-2-yl)amino)benzamide. Yield: 53.4%. MS(ESI)m / e317.0(M+H) + .

[0072] Step 2: Synthesis of compound 4-((4-(((3-(N-trideuteromethylmethanesulfonamide)pyrazine-2-yl)methyl)amino)-5-(trifluoromethyl)pyrimidine-2-yl)amino)benzamide (52) 5 mL of 1,2-dichloroethane and 5 mL of tert-butanol were added to a 25 mL round-bottom flask containing compound 52-2 prepared above, and the mixture was stirred at room temperature to dissolve and clarify. Next, compound 44-3 (63.0 mg, 0.20 mmol) and diisopropylethylamine (78 mg, 0.60 mmol) were added sequentially to the system, and the system was transferred to an oil bath at 80°C for reflux. The following day, TLC monitoring confirmed that the raw materials had been completely consumed. After stopping the heating and allowing the system to cool to room temperature, the solvent was removed by rotary evaporation to obtain the crude product, which was then separated and purified by pre-TLC to obtain 22 mg of the off-white solid 4-((4-(((3-(N-deuteromethylmethanesulfonamide)pyrazine-2-yl)methyl)amino)-5-(trifluoromethyl)pyrimidine-2-yl)amino))benzamide. Yield: 22.0%. MS(ESI)m / e500.1(M+H) + . 1 H NMR(400MHz,DMSO-d6)δ9.83(s,1H),8.69(s,2H),8.60(s,1H),8.31(s,1H),8.20(d,J=4.0Hz,1H), 7.68-7.60(dd,J=15.4,8.4Hz,4H),7.41-7.39(t,J=4.4Hz,1H),5.03(d,J=3.6Hz,2H),3.20(s,3H).

[0073] Compounds 48-50 and 53-55 were obtained by preparing compounds according to the method of the above examples using the known compounds 4-amino-3,5-diduterobenzoic acid and 4-amino-2,6-diduterobenzoic acid (Journal of Labelled Compounds and Radiopharmaceuticals, 53(11-12), 668-673; 2010) as raw materials. Compounds 59-61 were obtained by preparing compounds according to the method of the above examples using the known compound 4-amino-2,3,5,6-tetraduterobenzoic acid (Journal of Natural Products, 79(6), 1532-1537; 2016) as raw materials.

[0074] The following test examples demonstrate the beneficial effects of the present invention.

[0075] Test Example 1: Inhibitory activity of the deuterated compound of the present invention against FAK (1) Test method Inhibitory activity experiments on FAK enzyme were performed, referring to the method described in the literature (Cancer Res. 2008, 68, 1935). Specifically, the following procedures were followed: The test compound was diluted to 1000 nM, then serially diluted with DMSO in a 1:3 ratio. 0.1 L of the solution was placed in a 384-well plate, with two wells for each concentration. 5 L of 2x FAK enzyme solution was added, the mixture was centrifuged at 1000 rpm for 1 minute, and incubated at 25°C for 15 minutes. 5 L of 2x substrate solution was added and incubated at 25°C for 60 minutes. Furthermore, 5 L of Sa-XL665 solution and 5 μL of TK antibody-Eu3+ were added, the mixture was centrifuged at 1000 rpm for 1 minute, incubated at 25°C for 60 minutes, and finally, the fluorescence signal was read using an Envision 2104 plate reader to calculate the 50% inhibitory concentration IC50 of each compound against FAK enzyme. The known FAK inhibitor defactinib was used as a control.

[0076] (2) Test results Table 1 shows the inhibitory activity of each compound against FAK. The compounds produced by the present invention can effectively inhibit the activity of the FAK enzyme, and the deuterated compounds 41 and 45 of the present invention exhibit greater inhibitory activity against the FAK enzyme compared to the undeuterated compound defactinib. It was found to be more active.

[0077] [Table 1]

[0078] Test Example 2: Pharmacokinetic study of the deuterated compound of the present invention in rats (1) Test method A precise amount of the test drug (10 mg) was weighed out, and first 0.25 ml of N,N-dimethylacetamide (DMA) was added to dissolve it. Then, 0.5% sodium carboxymethylcellulose (CMC-Na) was slowly added up to 5 ml, and the mixture was homogeneously mixed using ultrasound or vortexing. 0.2 ml of the final solution prepared above was taken and stored at -20°C for concentration measurement.

[0079] Three healthy male adult SD rats (180-250g, purchased from Chengdu Dashuo Laboratory Animal Co., Ltd.) were fasted overnight (with free access to water) and then administered intragastricly at a dose of 5 ml / kg. 0.1 ml of blood was collected from the retroorbital venous plexus before administration and at 0.5, 1, 2, 4, 6, 8, 12, and 24 hours after administration. Plasma was separated by centrifugation at 4°C for 5 minutes and stored at -20°C for measurement. The concentration of the test compound in the plasma was measured by LC / MS / MS. The known FAK inhibitor defactinib was used as a control.

[0080] (2) Test results

[0081] [Table 2]

[0082] As shown in Table 2, the deuterated compounds 25 and 44 produced in the present invention exhibit superior pharmacokinetics compared to defactinib, with the compounds having higher peak blood concentration (Cmax), higher exposure (AUC), and longer half-lives. Therefore, the deuterated compounds produced by the present invention have even brighter future application potential among FAK inhibitors or drugs for treating cancer.

[0083] Test Example 3: Efficacy study in an animal tumor model using the deuterated compound of the present invention in combination with a PD-1 inhibitor. 1. MC38 tumor model: (1) Test method Cell culture: MC-38 cells were cultured in DMEM medium containing 10% fetal bovine serum (FBS). The cells were cultured in [specimen name]. MC-38 cells in the logarithmic growth phase were collected, resuspended in HBSS to the appropriate concentration, and used for subcutaneous tumor inoculation of C57BL / 6 mice.

[0084] Laboratory animals: C57BL / 6 mice, female, 6-8 weeks old, weighing approximately 18-20g, 96 individuals, purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.

[0085] Inoculation of tumor cells: Tumor cells in the logarithmic growth phase are collected, and the cell concentration is increased to 5 × 10⁻⁶ using HBSS. 6 The solution was adjusted to 5 × 10¹ / mL, and 0.1 mL was injected subcutaneously into the right side of each mouse's back using a 1 mL syringe. 5 The number of mice was determined to be per individual. Subsequently, the tumor volume was observed and measured, and the average tumor volume of the mice was 50-100 mm². 3 After growth, tumor-bearing mice were randomly divided into groups based on tumor volume and administered the treatment. Detailed information is shown in Table 3 below, with the day of group division and administration defined as day 0.

[0086] Tumor volume calculation: Mice were sacrificed on day 18, tumors were removed, and tumor volume was measured to calculate the tumor suppression rate for each group.

[0087] [Table 3]

[0088] (2) Test results Figure 1 shows the efficacy of the drug after 18 days of administration to animals, and Table 3 shows the calculated tumor suppression rate. It was found that the efficacy of compound 44 of the present invention alone was superior to that of the mPD-1 antibody alone, indicating that administration of the compound of the present invention alone has a therapeutic effect on mice with an MC38 tumor model.

[0089] Furthermore, when compound 44 was administered in combination with the mPD-1 antibody (group 5), the tumor suppression effect was significantly improved compared to when compound 44 was administered alone (group 4) or when the mPD-1 antibody was administered alone (group 2), demonstrating a synergistic effect.

[0090] Furthermore, when compound 44 of the present invention (50 mg / kg, administered once daily) was administered in combination with an mPD-1 antibody (Group 5), a higher and superior tumor suppression effect was obtained compared to when Defactinib (50 mg / kg, administered twice daily) was administered in combination with an mPD-1 antibody (Group 3). In other words, when used in combination with a PD-1 inhibitor, the compound of the present invention showed superior tumor suppression at half the dose of Defactinib. The observed effect demonstrates that the combination of the compound of the present invention with a PD-1 inhibitor exhibits significantly superior antitumor efficacy against the MC38 tumor model compared to the combination of defactinib and a PD-1 inhibitor.

[0091] 2. PAN02 Tumor Model (1) Test method PAN-02 cells in the logarithmic growth phase were collected, washed twice with PBS, and then resuspended in pre-cooled PBS for inoculation. The experimental animals were female C57BL / 6 mice, purchased from Beijing Weitong Lihua Laboratory Animals Co., Ltd. The C57BL / 6 mice were adapted to the laboratory environment for 3 days, and PAN-02 cells were subcutaneously inoculated into the right flank. The inoculation volume was 1 × 10⁶ cells. 6 The number was calculated per animal. The tumor was approximately 100 mm. 3 After growing to maturity, the animals were screened and randomly divided into groups of 8 each. They were then administered according to the group and administration plan shown in Table 4 below. The day of administration was defined as day 1, and the administration period was 33 days.

[0092] [Table 4]

[0093] (2) Test results Figure 2 shows the efficacy of the drug after 33 days of administration to animals, and Table 4 shows the calculated tumor suppression rates. When compound 44 was administered in combination with the mPD-1 antibody (group 5), the tumor suppression effect was significantly improved compared to when compound 44 was administered alone (group 4) or the mPD-1 antibody was administered alone (group 2).

[0094] On the other hand, when compound 44 of the present invention (25 mg / kg, administered twice daily) was administered in combination with an mPD-1 antibody (Group 5), the tumor suppression effect was significantly improved compared to when Defactinib (50 mg / kg, administered twice daily) was administered in combination with an mPD-1 antibody (Group 3). In other words, when used in combination with a PD-1 inhibitor, the compound of the present invention showed a superior tumor suppression effect at a lower dose than Defactinib. This indicates that the combination of the compound of the present invention and a PD-1 inhibitor has a significantly superior antitumor effect against the PAN-02 tumor model compared to the combination of Defactinib and a PD-1 inhibitor.

[0095] As described above, the present invention provides a deuterated compound that exhibits better pharmacokinetics, higher peak blood concentrations, higher exposure levels, and longer half-lives compared to the compound before deuteration, and has superior metabolic performance. Furthermore, the deuterated compound of the present invention can effectively inhibit the activity of FAK, and is useful in the preparation of FAK inhibitors and / or drugs for treating cancer. The deuterated compounds of this invention have great potential for applications in manufacturing. At the same time, when used in combination with anticancer drugs (e.g., PD-1 inhibitors), they can exert a synergistic effect, significantly improving tumor suppression and providing a superior option in clinical cancer treatment.

Claims

[Claim 1] Use of a concomitant drug in the preparation of a drug for treating cancer, The aforementioned combination drug comprises compound 44 and an anticancer drug to be administered simultaneously or separately. The aforementioned cancer is colon cancer or pancreatic cancer. The aforementioned anticancer drug is an mPD-1 antibody. 。

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