A pibk alpha / hdac6 isoform selective dual inhibitor and uses thereof

Abstract

The present application relates to a PI3K alpha / HDAC6 subtype selective dual inhibitor, which is a compound with general formula (I) as follows and pharmaceutically acceptable salts or solvates thereof: The present application also relates to the use of the PI3K alpha / HDAC6 subtype selective dual inhibitor in the preparation of antitumor drugs. The compound with general formula (I) of the present application is a subtype selective PI3K alpha / HDAC6 dual inhibitor, which has structural units required for inhibiting PI3K alpha and HDAC6, has significant PI3K alpha and HDAC6 dual inhibition activity, can avoid the shortcomings of PI3K and HDAC inhibitors and non-selective pan-PI3K / HDAC dual inhibitors in antitumor efficacy and toxicity, and can overcome the problems of drug-drug interactions, cumulative toxicity, complex pharmacokinetics, poor patient compliance and the like in the combination therapy of PI3K inhibitors and HDAC inhibitors, so the subtype selective PI3K alpha / HDAC6 dual inhibitor is particularly valuable in the treatment of proliferative diseases such as cancer.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceuticals, specifically relating to a selective dual inhibitor of PI3Kα / HDAC6 subtype and its application. Background Technology

[0002] The PI3K / Akt / mTOR signaling pathway, as an important intracellular signal transduction pathway, plays a vital biological role in cell growth, survival, proliferation, and apoptosis. Disruption of this pathway can lead to a range of diseases, including cancer, immune system disorders, and hematopoietic disorders. Related studies have shown that the key regulatory site PI3K in this pathway is closely related to tumorigenesis and development. The development of anti-tumor drugs targeting PI3K has become a research hotspot in recent years (Nature Reviews Molecular Cell Biology, 2012, 13: 195-203). Phosphatidylinositide 3-kinase (PI3K) is a class of lipid kinases comprising multiple members. Based on their structural characteristics, activation mechanisms, and selectivity for lipid substrates, PI3K kinases can be mainly divided into three classes: I, II, and III, with class I PI3K being the most thoroughly studied (Journal of Medicinal Chemistry, 2019, 62: 4815-4850). Class I PI3K kinases include four subtypes: PI3Kα, PI3Kβ, PI3Kδ, and PI3Kγ (Molecular Cancer, 2019, 18:26). Among them, PI3Kα is most closely associated with tumorigenesis and development. The main reason is that the PIK3CA gene, which encodes the catalytic subunit p110α, is one of the most easily mutated oncogenes. Mutations mainly occur at three sites: glutamate E542 and E545 on the helical domain PI3KC, and histidine H1047 on the kinase catalytic domain PI3Kc. Usually, glutamate E542 and E545 are mutated to lysine, while histidine H1047 is mutated to arginine. Tumors caused by these three mutation sites account for about 30% of all solid tumors (Science, 2004, 304:554). Mutations in PI3Kα can cause abnormal activation of the PI3K / Akt / mTOR signaling pathway, leading to excessive proliferation of tumor cells and the development of various malignant tumors, such as breast cancer, colon cancer, endometrial cancer, gastric cancer, ovarian cancer, and lung cancer (Aging, 2011, 3: 192-222). The other three subtypes, PI3Kβ, PI3Kδ, and PI3Kγ, mainly play roles in the formation of diseases such as thrombosis, immune dysfunction, leukemia, allergies, and inflammation (Journal of Medicinal Chemistry, 2019, 62: 4815-4850).Alpelisib (Norvartis) is currently the only successfully marketed selective inhibitor of the PI3Kα subtype. It was approved by the U.S. Food and Drug Administration (FDA) in 2019 for the treatment of postmenopausal women and men with HR+ / HER2- advanced or metastatic breast cancer who have disease progression during or after endocrine therapy and harbor PIK3CA gene mutations (Drugs, 2019, 79:1249-1253). However, this inhibitor has shown poor efficacy as a monotherapy, and therefore is currently mainly used in combination with fulvestrant in clinical practice.

[0003] Histone deacetylases (HDACs) are an important class of epigenetic enzymes that regulate gene expression by removing the ε-amino group from lysine residues on histones. HDACs can regulate the expression of various proteins, including tumor suppressor factors (p53, p21, etc.) and transcription factors (such as TFIIE, TCF, SF1, etc.) (Translational Oncology, 2022, 16: 101312). Dysregulation of HDACs is closely related to the occurrence and proliferation of cancer. HDAC inhibitors have become an effective method for cancer treatment, and a number of related inhibitors have been approved in recent years, including Voronostat (SAHA), Romidepsin, Belinostat, Panobinostat, and Chidamide (Bioorganic & Medicinal Chemistry Letters, 2021, 49: 128286). However, these small molecule inhibitors have produced many toxic side effects due to their lack of specificity and selectivity, such as hematological adverse reactions, gastrointestinal adverse reactions, and cardiotoxicity (Drug Safety, 2019, 42: 235-245). Notably, the HDAC6 subtype, which acts on the cytoplasm of the HDAC family, can regulate the acetylation status of various non-histone substrates (α-tubulin, HSP-90, and HSF-1, etc.) and the transformation, stress response, and metastasis of cancer cells, and is closely related to drug resistance (Bioorganic Chemistry, 2022, 127:105992). Furthermore, unlike the severe defects or lethal effects of knocking out class I HDACs, HDAC6 knockout mice can survive and remain healthy without significant phenotypic abnormalities (Neuropharmacology, 2016, 110:470-479). Therefore, targeting HDAC6 holds promise for better tolerability and efficacy. Currently, subtype-selective HDAC6 inhibitors, represented by Ricolinostat and Tubastain, are considered next-generation HDAC-targeting drugs (Bioorganic Chemistry, 2022, 127:105992).

[0004] A series of studies have shown that the combination of PI3K inhibitors and HDAC inhibitors can not only synergistically inhibit tumor growth, but also improve efficacy, limit drug resistance, and provide a better therapeutic window than single inhibitors (Cancer Cell, 2016, 29:311-323). Furthermore, given the problems associated with combination therapies, such as adverse drug interactions, cumulative toxicity, complex pharmacokinetics, and poor patient compliance (Anti-Cancer Agents in Medicinal Chemistry, 2019, 19:842-874), developing dual PI3K / HDAC inhibitors holds significant research value. This research not only promises superior efficacy compared to PI3K inhibitors, reduces drug resistance, and avoids the interactions, cumulative toxicity, complex pharmacokinetics, and poor patient compliance associated with combining PI3K and HDAC inhibitors, but also offers the potential for significant research benefits. Based on this, researchers both domestically and internationally have conducted numerous studies. However, to date, only the pan-PI3K / HDAC dual inhibitor CUDC-907 has entered clinical trials. Due to its inability to selectively inhibit specific PI3K subtypes, the inhibitor has exhibited toxicity and tolerability issues in clinical trials, causing it to remain in Phase II clinical trials (British Journal of Haematology, 2021, 195: 201-209). Summary of the Invention

[0005] On the one hand, this invention addresses the problems of existing PI3K and HDAC inhibitors, as well as non-selective pan-PI3K / HDAC dual inhibitors, in terms of antitumor efficacy and toxicity; on the other hand, it addresses the problems of drug interactions, cumulative toxic side effects, complex pharmacokinetics, and poor patient compliance associated with the combination therapy of PI3K inhibitors and HDAC inhibitors. Therefore, this invention provides a PI3Kα / HDAC6 subtype selective dual inhibitor and its application.

[0006] According to one aspect of the invention, the present invention provides a selective dual inhibitor of the PI3Kα / HDAC6 subtype, which is a compound having the general formula (I) and a pharmaceutically acceptable salt or solvate thereof:

[0007]

[0008] Where X, Y, and Z are all CH; or X and Y are CH and Z is N; or X and Z are CH and Y is N; or Y and Z are CH and X is N;

[0009] R1 is selected from C 1-6 Alkyl, fluorinated C 1-6 Alkyl, C 3-6 cycloalkyl, fluorinated C3-6 cycloalkyl, C 3-6 Heterocyclic alkyl or fluorinated C 3-6 Heterocyclic alkyl groups;

[0010] R2 is selected from hydrogen, halogens, and C. 1-6 Alkyl, fluorinated C 1-6 Alkyl, C 3-6 cycloalkyl, fluorinated C 3-6 cycloalkyl or C 3-6 Heterocyclic alkyl groups;

[0011] L is Ring A is a C ring that is replaced by at least one R3. 6-14 Aryl, C 5-14 The aromatic heterol group, R3, is independently selected from hydrogen, halogen, hydroxyl, cyano, carbamoyl, trifluoromethyl, trifluoromethoxy, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Unsaturated aliphatic hydrocarbon groups, N(R4)2, NR4OR4, NR4N(R4)2, SO2N(R4)2, NR4SO2R4, NR4CON(R4)2, NR4COOR4, NR4COR4, CON(R4)2, wherein R4 is independently selected from hydrogen, C 1-6 Alkyl or C 2-6 Unsaturated aliphatic hydrocarbon group.

[0012] Activity assays confirmed that the compound of general formula (I) of this invention is a subtype-selective dual inhibitor of PI3Kα / HDAC6, possessing the structural units required to inhibit both PI3Kα and HDAC6, and exhibiting significant dual inhibitory activity against both PI3Kα and HDAC6. Specifically, the compound of general formula (I) of this invention has a key site of action with PI3Kα to provide selective inhibition of PI3Kα, and the introduced aromatic ring or aromatic heterocyclic hydroxamic acid structure can interact more effectively with the catalytic channel and zinc ion cofactor of HDAC6 to provide selective inhibition of HDAC6. Compared to CUDC-907, the compound of general formula (I) of this invention inhibits fewer than 10 targets of the PI3K and HDAC families from two targets closely related to tumor development and progression, thus exhibiting lower toxicity and a larger tolerable dose (therapeutic window). In addition, due to the single-molecule dual-target effect, the compound of the present invention with general formula (I) can avoid the problems of drug-to-drug interactions, cumulative toxic side effects, complex pharmacokinetics, and poor patient compliance in the combination therapy of PI3K inhibitors and HDAC inhibitors, and is particularly valuable in the treatment of proliferative diseases such as cancer.

[0013] The compounds having general formula (I) in this invention can exist as hydrates, solvates, polymorphs, and mixtures thereof.

[0014] The compounds having general formula (I) in this invention may exist in an optically active form, a mixture of optical isomers, one or more tautomers and mixtures thereof, or one or more tautomers and mixtures thereof.

[0015] Preferably, R1 is selected from

[0016] Preferably, R2 is selected from methyl or chlorine.

[0017] Preferably, the compound having general formula (I) is selected from any of the compounds numbered 1-19:

[0018]

[0019]

[0020] It should be noted that the compounds numbered 1-19 above are only examples to make the technical solution of the present invention clearer. The PI3Kα / HDAC6 subtype selective dual inhibitor of the present invention includes not only the compounds numbered 1-19 above and their pharmaceutically acceptable salts or solvates.

[0021] According to another aspect of the invention, the invention provides the use of a PI3Kα / HDAC6 subtype selective dual inhibitor in the preparation of an antitumor drug.

[0022] Preferably, the tumor includes a solid tumor or a hematoma.

[0023] The indications for this invention—tumors—include, but are not limited to, lung and bronchial cancer; prostate cancer; breast cancer; pancreatic cancer; colon and rectal cancer; thyroid cancer; liver and intrahepatic bile duct cancer; hepatocellular carcinoma; gastric cancer; glioma / glioblastoma; endometrial cancer; melanoma; kidney and renal pelvis cancer; bladder cancer; endometrial cancer; cervical cancer; ovarian cancer; multiple myeloma; esophageal cancer; acute myeloid leukemia; chronic myeloid leukemia; lymphocytic leukemia; myeloid leukemia; brain cancer; oral and pharyngeal cancer; laryngeal cancer; small bowel cancer; non-Hodgkin lymphoma; melanoma; and villous colonic adenoma, etc.

[0024] Preferably, the dosage of the PI3Kα / HDAC6 subtype selective dual inhibitor is 1 mg to 1000 mg / day. It should be understood that this dosage is for illustrative purposes only and is not a limitation, and may deviate from this range depending on the severity of the condition or the dosage form.

[0025] Preferably, the PI3Kα / HDAC6 subtype selective dual inhibitor functions in the form of a composition.

[0026] In this invention, "combination" refers to a fixed combination in the form of dosage units, or a kit for combined administration, wherein a compound having general formula (I) and combination partners (such as other drugs) can be administered simultaneously and independently or separately at time intervals, particularly when these time intervals allow the combination partners to exhibit synergistic effects. This administration provides therapeutically effective levels of two or more drugs (compounds) in the patient (e.g., cocktail therapy).

[0027] Preferably, the composition further includes at least one pharmaceutical carrier or excipient.

[0028] Preferably, the composition further includes at least one anticancer agent.

[0029] This invention provides a method for treating proliferative diseases such as cancer, comprising administering, alone or in combination with one or more other anticancer agents, a therapeutically effective amount of a compound having general formula (I). Specifically, the components are formulated together as a combination therapy or administered separately. Anticancer agents suitable for use with compounds having general formula (I) include, but are not limited to, one or more compounds selected from kinase inhibitors (such as gefitinib, erlotinib, etc.), anti-estrogens (such as fulvestrant, tamoxifen, toremifene, raloxifene, anastrozole, etc.), anti-androgens (such as flutamide, bicalutamide, finasteride, aminoglutethimide, ketoconazole, and corticosteroids, etc.), cancer chemotherapy drugs, alkylating agents, chelating agents, biological effector modifiers, cancer vaccines, and compounds of substances for antisense therapy.

[0030] Preferably, the dosage form of the composition is clinically or pharmaceutically acceptable.

[0031] In summary, the subtype-selective PI3Kα / HDAC6 dual inhibitors involved in this invention can provide a new therapy for anti-tumor treatment. Multiple experiments have confirmed that the compounds of general formula (I) provided by this invention can strongly inhibit PI3Kα and HDAC6, while exhibiting excellent PI3Kα / HDAC6 subtype selectivity, and have the potential to be developed into anti-tumor drugs. Detailed Implementation

[0032] Unless otherwise specified, the term "alkyl" as used in this invention refers to a straight or branched hydrocarbon chain consisting of 1 to 6 carbon atoms.

[0033] Unless otherwise specified, the term "cycloalkyl" as used in this invention refers to monocycloalkyl, spirocycloalkyl, and bridged cycloalkyl with 3 to 11 carbon atoms, which may or may not contain alkyl substituents.

[0034] Unless otherwise specified, the term "heterocyclic alkyl" as used in this invention refers to monocyclic heteroalkyl, spirocyclic heteroalkyl, and bridged heteroalkyl compounds consisting of 2-9 carbon atoms and 1-2 heteroatoms (e.g., oxygen, nitrogen, sulfur, etc.), which may or may not contain alkyl substituents.

[0035] As used in this invention, the term "halogen" refers to fluorine, bromine, chlorine, or iodine, particularly fluorine and chlorine.

[0036] The term "fluorinated" as used in this invention means that fluorinated alkyl, cycloalkyl, or heterocycloalkyl groups can be mono-, poly-, or fully halogenated.

[0037] The term "aryl" as used in this invention refers to an all-carbon monocyclic or fused polycyclic group with 5-12 carbon atoms and a fully conjugated π-electron system.

[0038] The term "heteroaryl" as used in this invention refers to a non-all-carbon monocyclic or fused polycyclic group with 5-12 carbon atoms and a fully conjugated π-electron system.

[0039] In this invention, "pharmaceutically acceptable salts" include inorganic acid salts, lower alkyl sulfonates, aryl sulfonates, organic acid salts, amino acid salts, etc.

[0040] In this invention, "treatment" includes preventive (protective) and therapeutic treatments as well as delaying the progression of a disease or disorder.

[0041] The compounds having the general structural formula (I) provided by the present invention are prepared by the following steps, but are not limited to the following methods.

[0042] Synthesis of intermediate B

[0043]

[0044] A 2-aminothiazole derivative (starting material A) undergoes a substitution reaction with phenyl chloroformate to yield a carbamate compound (intermediate B). The base used in the reaction is diisopropylethylamine (DIPEA), the solvent is 1,4-dioxane, the reaction temperature is 40°C, and the reaction time is 12 hours.

[0045] The following is an example of synthetic intermediate B.

[0046] Synthesis of intermediate B-1

[0047] Phenyl(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamate

[0048]

[0049] At room temperature, 4-methyl-5-(2-(1,1,1,-trifluoro-2-methylpropyl-2-yl)pyridin-4-yl)thiazole (250 mg, 0.83 mmol) and DIPEA (360 μL, 2.07 mmol) were dissolved in anhydrous 1,4-dioxane. The temperature was raised to 40 °C, and phenyl chloroformate (156 μL, 1.24 mmol) was slowly added dropwise. The reaction was continued for 12 hours. After the reaction was complete, the solvent was removed by vacuum distillation, and then the mixture was dissolved in ethyl acetate, washed twice with 1N sodium hydroxide, and the organic layer was collected and dried over anhydrous sodium sulfate. The solvent was removed by vacuum distillation to obtain the crude product. The crude product was purified by column chromatography to obtain 150 mg of an off-white solid, yield: 45%. 1H NMR (400MHz, DMSO-d6) δ12.53 (s, 1H, NH), 8.63 (d, J=5.1Hz, 1H, Ar-H), 7.59 (s, 1H, Ar-H), 7.50-7.41 ( m, 3H, Ar-H), 7.36-7.21 (m, 3H, Ar-H), 2.44 (s, 3H, CH3), 1.62 (s, 6H, CH3×2); ESI-MS: m / z=422[M+H]+.

[0050] Synthesis of intermediate B-2

[0051] Phenyl(5-(2-(tert-butyl)pyridin-4-yl)-4-methylthiazolyl-2-yl)carbamate

[0052]

[0053] The synthetic method was the same as that for intermediate B-1, except that 4-methyl-5-(2-(1,1,1,-trifluoro-2-methylpropyl-2-yl)pyridin-4-yl)thiazole was replaced with 5-(2-(tert-butyl)pyridin-4-yl)-4-methylthiazole-2-amine, yielding 120 mg of an off-white solid, yield: 47%. ESI-MS: m / z = 368 [M+H]+.

[0054] Synthesis of intermediate B-3

[0055] Phenyl(5-(2-cyclobutylpyridin-4-yl)-4-methylthiazolyl-2-yl)carbamate

[0056]

[0057] The synthetic method was the same as that for intermediate B-1, except that 4-methyl-5-(2-(1,1,1,-trifluoro-2-methylpropyl-2-yl)pyridin-4-yl)thiazole was replaced with 5-(2-cyclobutylpyridin-4-yl)-4-methylthiazole-2-amine, yielding 132 mg of an off-white solid, yield: 51%. ESI-MS: m / z = 366 [M+H]+.

[0058] Synthesis of intermediate B-4

[0059] Phenyl(4-methyl-5-(2-(1-methylcyclopropyl)pyridin-4-yl)thiazolyl-2-yl)carbamate

[0060]

[0061] The synthetic method was the same as that for intermediate B-1, except that 4-methyl-5-(2-(1,1,1,-trifluoro-2-methylpropyl-2-yl)pyridin-4-yl)thiazole was replaced with 4-methyl-5-(2-(1-methylcyclopropyl)pyridin-4-yl)thiazole-2-amine, yielding 126 mg of an off-white solid, yield: 48%. ESI-MS: m / z = 366 [M+H]+.

[0062] Synthesis of intermediate B-5

[0063] Phenyl(5-(2-(1-cyanocyclopropyl)pyridin-4-yl)-4-methylthiazolyl-2-yl)carbamate

[0064]

[0065] The synthetic method was the same as that for intermediate B-1, except that 4-methyl-5-(2-(1,1,1,-trifluoro-2-methylprop-2-yl)pyridin-4-yl)thiazole was replaced with 1-(4-(2-amino-4-methylthiazolin-5-yl)pyridin-2-yl)cyclopropane-1-onitrile, yielding 106 mg of an off-white solid, yield: 58%. ESI-MS: m / z = 377 [M+H]+.

[0066] Synthesis of intermediate B-6

[0067] Phenyl(4-methyl-5-(2-(1-(trifluoromethyl)cyclopropyl)pyridin-4-yl)thiazolyl)carbamate

[0068]

[0069] The synthetic method was the same as that for intermediate B-1, except that 4-methyl-5-(2-(1,1,1,-trifluoro-2-methylpropyl-2-yl)pyridin-4-yl)thiazole was replaced with 4-methyl-5-(2-(1-(trifluoromethyl)cyclopropyl)pyridin-4-yl)thiazole-2-amine, yielding 125 mg of an off-white solid, yield: 47%. ESI-MS: m / z = 420 [M+H]+.

[0070] Synthesis of intermediate B-7

[0071] Phenyl(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyrimidin-4-yl)thiazolyl-2-yl)carbamate

[0072]

[0073] The synthetic method was the same as that for intermediate B-1, except that 4-methyl-5-(2-(1,1,1,-trifluoro-2-methylpropyl-2-yl)pyridin-4-yl)thiazole was replaced with 4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyrimidin-4-yl)thiazole-2-amine, yielding 118 mg of an off-white solid, yield: 51%. ESI-MS: m / z = 423 [M+H]+.

[0074] Synthesis of intermediate B-8

[0075] Phenyl(5-(2-(tert-butyl)pyrimidin-4-yl)-4-methylthiazolyl-2-yl)carbamate

[0076]

[0077] The synthetic method was the same as that for intermediate B-1, except that 4-methyl-5-(2-(1,1,1,-trifluoro-2-methylpropyl-2-yl)pyridin-4-yl)thiazole was replaced with 5-(2-(tert-butyl)pyrimidin-4-yl)-4-methylthiazole-2-amine, yielding 145 mg of an off-white solid, yield: 55%. ESI-MS: m / z = 369 [M+H]+.

[0078] Synthesis of intermediate B-9

[0079] Phenyl(5-(2-cyclopropylpyrimidin-4-yl)-4-methylthiazolyl-2-yl)carbamate

[0080]

[0081] The synthetic method was the same as that for intermediate B-1, except that 4-methyl-5-(2-(1,1,1,-trifluoro-2-methylpropyl-2-yl)pyridin-4-yl)thiazole was replaced with 5-(2-cyclopropylpyrimidin-4-yl)-4-methylthiazole-2-amine, yielding 135 mg of an off-white solid, yield: 51%. ESI-MS: m / z = 353 [M+H]+.

[0082] Synthesis of intermediate B-10

[0083] Phenyl(4-methyl-5-(2-(1-methylcyclopropyl)pyrimidin-4-yl)thiazolyl-2-yl)carbamate

[0084]

[0085] The synthetic method was the same as that for intermediate B-1, except that 4-methyl-5-(2-(1,1,1,-trifluoro-2-methylpropyl-2-yl)pyridin-4-yl)thiazole was replaced with 4-methyl-5-(2-(1-methylcyclopropyl)pyrimidin-4-yl)thiazole-2-amine, yielding 128 mg of an off-white solid, yield: 52%. ESI-MS: m / z = 367 [M+H]+.

[0086] Synthesis of intermediate B-11

[0087] Phenyl(5-(2-(azacyclobutan-1-yl)pyrimidin-4-yl)-4-methylthiazolyl-2-yl)carbamate

[0088]

[0089] The synthetic method was the same as that for intermediate B-1, except that 4-methyl-5-(2-(1,1,1,-trifluoro-2-methylpropyl-2-yl)pyridin-4-yl)thiazole was replaced with 5-(2-(azacyclobutan-1-yl)pyrimidin-4-yl)-4-methylthiazole-2-amine, yielding 133 mg of an off-white solid, yield: 49%. ESI-MS: m / z = 368 [M+H]+.

[0090] Synthesis of intermediate B-12

[0091] Phenyl(5-(2-(diethylamino)pyrimidin-4-yl)-4-methylthiazolyl-2-yl)carbamate

[0092]

[0093] The synthetic method was the same as that for intermediate B-1, except that 4-methyl-5-(2-(1,1,1,-trifluoro-2-methylpropyl-2-yl)pyridin-4-yl)thiazole was replaced with 5-(2-(diethylamino)pyrimidin-4-yl)-4-methylthiazole-2-amine, yielding 111 mg of an off-white solid, yield: 32%. ESI-MS: m / z = 384 [M+H]+.

[0094] Synthesis of intermediate B-13

[0095] Phenyl(4-chloro-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamate

[0096]

[0097] The synthetic method was the same as that for intermediate B-1, except that 4-methyl-5-(2-(1,1,1,-trifluoro-2-methylpropyl-2-yl)pyridin-4-yl)thiazole was replaced with 4-chloro-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazole-2-amine, yielding 127 mg of an off-white solid, yield: 52%. ESI-MS: m / z = 442 [M+H]+.

[0098] Synthesis of intermediate D

[0099]

[0100] (4R)-amino-L-proline amide (starting material C) protected by tert-butyloxycarbonyl group undergoes an acid-ammonia condensation reaction or a nucleophilic substitution reaction with a derivative containing an ester group to yield a compound containing an ester group (intermediate D). The base used in the reaction is diisopropylethylamine (DIPEA), the solvent is dichloromethane (DCM) or isopropyl alcohol, the reaction temperature is room temperature or 130°C, and the reaction time is 4 hours or 12 hours.

[0101] The following is an example of the synthesis intermediate D.

[0102] Synthesis of intermediate D-1

[0103] (2S,4R)-2-carbamoyl-4-(4-(methoxycarbonyl)benzoylamino)pyrrolidine-1-carboxylic acid tert-butyl ester

[0104]

[0105] Monomethyl terephthalate (108 mg, 0.60 mmol), HOBT (81 mg, 0.60 mmol), and EDCI (173 mg, 0.90 mmol) were dissolved in anhydrous dichloromethane at room temperature and reacted for 2 hours. DIPEA (263 μL, 1.5 mmol) was added to the solution at room temperature, and after 10 minutes, starting material C (150 mg, 0.66 mmol) was added at room temperature. The reaction was continued at room temperature for 3 hours until the reaction was complete. The reaction mixture was washed twice with saturated sodium bicarbonate, and the organic layer was collected and concentrated under vacuum. The resulting product was purified by column chromatography (4% CH3OH / CH2Cl2) to obtain a pale yellow solid (129 mg, 0.33 mmol), yield: 55%. 1 H NMR (400MHz, DMSO-d6) δ8.78-8.71 (m, 1H, NH), 8.03 (d, J=8.3Hz, 2H, Ar-H), 7.99-7.92 (m , 2H, Ar-H), 7.50-7.31 (m, 1H, NH), 7.10-6.90 (m, 1H, NH), 4.61-4.43 (m, 1H, CH), 4.25-4.1 2(m,1H,CH),3.87(s,3H,CH3),3.75-3.64(m,1H,CH),3.34-3.22(m,1H,CH),2.31-2.13( m, 1H, CH), 2.15-2.02 (m, 1H, CH), 1.38 (s, 4H, CH), 1.35 (s, 5H, CH); ESI-MS: m / z=392[M+H] + .

[0106] Synthesis of intermediate D-2

[0107] (2S,4R)-2-carbamoyl-4-(2-(4-(methoxycarbonyl)phenyl)acetamido)pyrrolidine-1-carboxylic acid tert-butyl ester

[0108]

[0109] The synthesis method was the same as that for intermediate D-1, except that monomethyl terephthalate was replaced with 4-(methoxycarbonyl)phenylacetic acid, yielding 131 mg of off-white solid, yield: 33%. 1H NMR (400MHz, DMSO-d6) δ8.44-8.36 (m, 1H, NH), 7.90 (d, J=8.2Hz, 2H, Ar-H), 7.39 (d, J=8. 1Hz, 2H, Ar-H), 7.37-7.34 (m, 1H, NH), 7.09-6.89 (m, 1H, NH), 4.34-4.17 (m, 1H, CH), 4.19 -4.04(m,1H,CH),3.84(s,3H,CH3),3.63-3.53(m,1H,CH),3.50(s,2H,CH2),3.19-3.06( m, 1H, CH), 2.17-1.75 (m, 2H, CH), 1.38 (s, 4H, CH), 1.35 (s, 5H, CH); ESI-MS: m / z=406[M+H] + .

[0110] Synthesis of intermediate D-3

[0111] (2S,4R)-2-carbamoyl-4-(5-(methoxycarbonyl)pyridine-2-acylamino)pyrrolidine-1-carboxylic acid tert-butyl ester

[0112]

[0113] The synthesis method was the same as that for intermediate D-1, except that monomethyl terephthalate was replaced with 5-(methoxycarbonyl)-2-pyridinecarboxylic acid, yielding 120 mg of a pale yellow solid, with a yield of 38%. 1 H NMR (400MHz, DMSO-d6) δ9.15-9.04 (m, 2H, Ar-H), 8.52-8.42 (m, 1H, NH), 8.21-8.11 (m, 1H, Ar-H), 7.47-7.36 (m, 1H, NH), 7.07-6.94 (m, 1H, NH), 4.71-4.50 (m, 1H, CH), 4.28-4.08 (m, 1H, CH), 3.93 (s, 3H, CH3), 3.72-3.53 (m, 2H, CH), 2.38-2.25 (m, 1H, C H), 2.07-1.96 (m, 1H, CH), 1.38 (s, 4H, CH), 1.35 (s, 5H, CH); ESI-MS: m / z=393[M+H] + .

[0114] Synthesis of intermediate D-4

[0115] 6-(((3R,5S)-1-(tert-butoxycarbonyl)-5-carbamoylpyrrolidine-3-yl)amino)nicotinic acid methyl ester

[0116]

[0117] Raw material C (500 mg, 2.18 mmol) and methyl 6-chloronicotinic acid (451 mg, 2.62 mmol) were dissolved in isopropanol. DIPEA (1.2 mL, 6.55 mmol) was added to the solution at room temperature, and the temperature was raised to 130 °C. The reaction was continued for 12 hours. After the reaction was completed, the solvent was removed by vacuum distillation to obtain the crude product. The crude product was purified by column chromatography to obtain a pale yellow solid (76 mg, 0.21 mmol), yield: 10%. 1 H NMR (400MHz, DMSO-d6) δ8.57 (s, 1H, Ar-H), 7.81 (d, J=8.9Hz, 1H, Ar-H), 7.68-7.61 (m, 1 H, Ar-H), 7.44-7.38 (m, 1H, NH), 7.04-6.95 (m, 1H, NH), 6.55-6.47 (m, 1H, NH), 4.57-4.37 (m, 1H, CH), 4.19-4.08 (m, 1H, CH), 3.75 (s, 3H, CH3), 3.73-3.63 (m, 1H, CH), 3.22-3.09 (m , 1H, CH), 2.17-2.04 (m, 2H, CH), 1.37 (s, 4H, CH), 1.35 (s, 5H, CH); ESI-MS: m / z=365[M+H] + .

[0118] Synthesis of intermediate D-5

[0119] 2-(((3R,5S)-1-(tert-butoxycarbonyl)-5-carbamoylpyrrolidine-3-yl)amino)pyrimidine-5-carboxylic acid methyl ester

[0120]

[0121] The synthesis method is the same as that for intermediate D-4, except that methyl 6-chloronicotinic acid is replaced with methyl 2-chloropyrimidine-5-carboxylic acid, yielding 146 mg of pale yellow solid, with a yield of 50%. 1H NMR (400MHz, DMSO-d6) δ8.81-8.75 (m, 1H, Ar-H), 8.74-8.68 (m, 1H, Ar-H), 8.41-8. 33 (m, 1H, NH), 7.45-7.35 (m, 1H, NH), 7.05-6.90 (m, 1H, NH), 4.63-4.43 (m, 1H, CH), 4.19-4.06 (m, 1H, CH), 3.77 (s, 3H, CH3), 3.72-3.64 (m, 1H, CH), 3.25-3.14 (m, 1H, C H), 2.20-2.00 (m, 2H, CH), 1.36 (s, 4H, CH), 1.33 (s, 5H, CH); ESI-MS: m / z=366[M+H] + .

[0122] Synthesis of intermediate D-6

[0123] (2S,4R)-2-formamido-4-((4-(methoxycarbonyl)phenyl)sulfonylamino)pyrrolidine-1-tert-butyl carboxylate

[0124]

[0125] At room temperature, starting material C (150 mg, 0.66 mmol) was dissolved in anhydrous dichloromethane, and then methyl 4-(chlorosulfonyl)benzoate (140 mg, 0.60 mmol) was slowly added dropwise under ice bath conditions. After reacting at room temperature for two hours, crude product was obtained. The crude product was purified by silica gel chromatography to obtain an off-white solid (130 mg, 0.30 mmol), yield: 50%. 1 H NMR (400MHz, DMSO-d6) δ8.32-8.23 (m, 1H, NH), 8.17 (d, J=8.2Hz, 2H, Ar-H), 7.95 (d, J=8. 2Hz, 2H, Ar-H), 7.41-7.29(m, 1H, NH), 6.99-6.86(m, 1H, NH), 4.08-3.99(m, 1H, CH), 3.90( s, 3H, CH3), 3.79-3.68 (m, 1H, CH), 3.41-3.34 (m, 1H, CH), 3.03-2.90 (m, 1H, CH), 2.04-1.8 8(m, 1H, CH), 1.87-1.73 (m, 1H, CH), 1.31 (d, J=3.28Hz, 9H, CH3×3); ESI-MS: m / z=428[M+H] + .

[0126] Synthesis of intermediate D-7

[0127] (2S,4R)-2-formamido-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)pyrrolidine-1-tert-butyl carboxylate)

[0128]

[0129] The synthesis method is the same as that for intermediate D-6, except that methyl 4-(chlorosulfonyl)benzoate is replaced with methyl 4-((chlorosulfonyl)methyl)benzoate, yielding 120 mg of off-white solid, yield: 45%. 1 H NMR (400MHz, DMSO-d6) δ7.96 (d, J=7.9Hz, 2H, Ar-H), 7.57-7.54 (m, 1H, NH), 7.52 (d, J=8. 1Hz, 2H, Ar-H), 7.41-7.28(m, 1H, NH), 7.01-6.87(m, 1H, NH), 4.48(s, 2H, CH2), 4.12-4.0 2(m,1H,CH), 3.95-3.88(m,1H,CH), 3.86(s,3H,CH3), 3.64-3.51(m,1H,CH), 3.19-3.03( m, 1H, CH), 2.11-1.94 (m, 2H, CH), 1.38 (s, 4H, CH), 1.34 (s, 5H, CH); ESI-MS: m / z=442[M+H] + .

[0130] Synthesis of intermediate F

[0131]

[0132] Intermediate D was hydrolyzed in the presence of trifluoroacetic acid (TFA) to give intermediate E. The solvent used was dichloromethane (DCM), the reaction temperature was from 0 degrees to room temperature, and the reaction time was 2 hours.

[0133]

[0134] Intermediate E and intermediate B undergo a substitution reaction under alkaline conditions to yield intermediate F. The base used in the reaction is diisopropylethylamine (DIPEA), the solvent is 1,4-dioxane, the reaction temperature is 60℃, and the reaction time is 20 hours.

[0135] The following is an example of synthetic intermediate F.

[0136] Synthesis of intermediate F-1

[0137] 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)methyl benzoate

[0138]

[0139] Dissolve the intermediate (2S,4R)-2-carbamoyl-4-(4-(methoxycarbonyl)benzoylamino)pyrrolidine-1-carboxylic acid tert-butyl ester in anhydrous dichloromethane, add an equal volume of trifluoroacetic acid dropwise under ice bath stirring, react at room temperature for 2 hours, and remove the solvent by vacuum distillation to obtain the deBoc product (intermediate E-1).

[0140] Intermediate phenyl(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazo-2-yl)carbamate (72 mg, 0.17 mmol) and DIPEA (42 μL, 0.17 mmol) were dissolved in anhydrous 1,4-dioxane at room temperature. The solution was heated to 60 °C, and intermediate E-1 (44 mg, 0.17 mmol) was slowly added dropwise. The reaction was carried out for 12 hours. After the reaction was completed, the solvent was removed by vacuum distillation to obtain the crude product. The crude product was purified by silica gel chromatography (6% CH3OH / CH2Cl2) to give an off-white solid (50 mg, 0.08 mmol), yield: 47%. 1 H NMR (400MHz, DMSO-d6) δ 11.18 (s, 1H, NH), 8.85 (d, J = 5.9Hz, 1H, Ar-H), 8.60 (d, J = 5.0Hz, 1H, Ar-H), 8.04 ( d, J=8.2Hz, 2H, Ar-H), 7.97 (d, J=8.2Hz, 2H, Ar-H), 7.55 (s, 1H, NH), 7.48 (s, 1H, NH), 7.41 (d, J=4.9Hz, 1H, Ar-H), 7.04 (s, 1H, NH), 4.67-4.53 (m, 1H, CH), 4.47-4.33 (m, 1H), 3.95-3.90 (m, 1H, CH), 3.88 (s, 3H, CH3), 3.60-3.47 (m, 1H, CH), 2.41 (s, 3H, CH3), 2.34-2.20 (m, 1H, CH), 2.18-2.03 (m, 1H, CH), 1.61 (s, 6H, CH3×2); 13C NMR (100MHz, DMSO-d6) δ174.01, 169.86, 167.79, 158.58, 149.36, 141.05, 136.87, 135.01, 133.05, 130.98, 129.66, 128.98, 128.02, 127.69, 124.08, 123.12, 121.59, 120.47, 60.32, 51.62, 450.36, 48.79, 46.74 (q, J c-F =24.0Hz), 35.42, 21.76, 21.75, 21.74, 16.76; ESI-MS: m / z=619[M+H] + .

[0141] Synthesis of intermediate F-2

[0142] 4-(2-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl)amino)-2-oxoethyl)methyl benzoate

[0143]

[0144] The synthesis method was the same as that for intermediate F-1, yielding 56 mg of off-white solid, with a yield of 52%. 1 H NMR (400MHz, DMSO-d6) δ11.05 (s, 1H, NH), 8.61 (d, J=5.1Hz, 1H, Ar-H), 8.54-8.46 (m, 1H, NH), 7 .87 (d, J=8.0Hz, 2H, Ar-H), 7.56 (s, 1H, Ar-H), 7.54-7.43 (m, 1H, NH), 7.43-7.36 (m, 3H, Ar-H), 7 .10-6.96 (m, 1H, NH), 4.50-4.27 (m, 2H, CH), 3.81 (s, 3H, CH3), 3.77-3.69 (m, 1H, CH), 3.54-3.48 (m, 2H, CH), 3.46-3.38 (m, 1H, CH), 2.41 (s, 3H, CH3), 2.15-1.94 (m, 2H, CH), 1.62 (s, 6H, CH3×2); 13C NMR (150MHz, DMSO-d6) δ174.31, 169.69, 168.52, 165.01, 159.96, 158.25, 155.34, 149.67, 146.98, 143.79, 141.36, 140.79, 134.59, 130.06, 128.09, 126.08, 121.59, 120.49, 118.62, 59.08, 51.59, 49.85, 46.79 (q, J C-F =24.0Hz), 35.75, 29.79, 21.93, 21.91, 21.89, 16.43; ESI-MS: m / z=633[M+H] + .

[0145] Synthesis of intermediate F-3

[0146] (2S,4R)-4-(5-(methoxycarbonyl)pyridine-2-acylamino)-N 1 -(4-Methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0147]

[0148] The synthesis method was the same as that for intermediate F-1, yielding 52 mg of a pale yellow solid, with a yield of 49%. 1 H NMR ((600MHz, DMSO-d6) δ 11.06 (s, 1H, NH), 9.31-9.22 (m, 1H, NH), 9.11 (d, J = 1.5Hz, 1H, Ar-H), 8.60 (d, J = 5.2Hz, 1H, Ar-H), 8.49 (dd, J=8.1, 2.1Hz, 1H, Ar-H), 8.18 (d, J=8.2Hz, 1H, Ar-H), 7.56 (s, 1H, Ar-H), 7.52-7.47 (m, 1H, NH), 7.42 (d, J=4.1Hz, 1 H, Ar-H), 7.11-7.01 (m, 1H, NH), 4.78-4.64 (m, 1H, CH), 4.45-4.32 (m, 1H, CH), 3.92 (s, 3H, CH3), 3.91-3.85 (m, 1H, CH), 3.61 -3.53 (m, 1H, CH), 2.41 (s, 3H, CH3), 2.39-2.32 (m, 1H, CH), 2.11-1.98 (m, 1H, CH), 1.62 (s, 6H, CH3×2); ESI-MS: m / z=620[M+H] + .

[0149] Synthesis of intermediate F-4

[0150] 2-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl)amino)pyridine-5-carboxylic acid methyl ester

[0151]

[0152] The synthesis method was the same as that for intermediate F-1, yielding 30 mg of a yellowish solid, with a yield of 45%. 1 H NMR (400MHz, CDCl3-d6) δ8.62 (s, 1H, NH), 8.57-8.41 (m, 1H, Ar-H), 7.81 (d, J=6.7Hz, 1H, Ar-H), 7 .67-7.35 (m, 2H, Ar-H), 7.33-7.24 (m, 1H, Ar-H), 7.20 (s, 1H, NH), 7.09 (s, 1H, NH), 6.36 (d, J=6.3 Hz, 1H, Ar-H), 6.20 (s, 1H, NH), 4.80-4.66 (m, 1H, CH), 4.64-4.50 (m, 1H, CH), 4.18-3.98 (m, 1H, CH) ), 3.74 (s, 3H, CH3), 3.62 (s, 1H, CH), 2.62-2.44 (m, 1H, CH), 2.27 (s, 4H, CH), 1.57 (s, 6H, CH3×2); 113 C NMR (101MHz, CDCl3-d6) δ175.15, 158.59, 155.46, 149.35, 148.75, 141.29, 135.45, 131.05, 12 9.98, 127.58, 125.34, 121.05, 120.79, 115.28, 110.03, 58.79, 55.45, 51.05, 48.72, 46.73 (q, J C-F =24.0Hz), 35.49, 21.89, 21.87, 21.86, 16.73; ESI-MS: m / z=592[M+H] + .

[0153] Synthesis of intermediate F-5

[0154] 2-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl)amino)pyrimidine-5-carboxylic acid methyl ester

[0155]

[0156] The synthesis method was the same as that for intermediate F-1, yielding 39 mg of a yellowish solid, with a yield of 55%. 1 H NMR (400MHz, DMSO-d6) δ11.03 (s, 1H, NH), 8.84-8.71 (m, 2H, Ar-H), 8.60 (d, J=5.1Hz, 1H, Ar-H) , 8.51-8.44 (m, 1H, NH), 7.55 (s, 1H, Ar-H), 7.49-7.44 (m, 1H, NH), 7.41 (d, J=4.7Hz, 1H, Ar-H), 7 .07-6.98 (m, 1H, NH), 4.72-4.59 (m, 1H, CH), 4.48-4.31 (m, 1H, CH), 3.96-3.86 (m, 1H, CH), 3.80 ( s, 3H, CH3), 3.55-3.42 (m, 1H, CH), 2.40 (s, 3H, CH3), 2.28-2.04 (m, 2H, CH), 1.62 (s, 6H, CH3×2); 13 C NMR (100MHz, DMSO-d6) δ173.28, 165.15, 159.03, 158.59, 156.25, 149.36, 141.48, 131.06 , 129.51, 128.31, 125.43, 121.58, 120.63, 116.28, 60.26, 56.02, 52.65, 49.69, 46.75 (q, J C-F =24.0Hz), 34.59, 21.85, 21.84, 21.82, 16.74; ESI-MS: m / z=593[M+H] + .

[0157] Synthesis of intermediate F-6

[0158] (2S,4R)-4-((4-(methoxycarbonyl)phenyl)sulfonylamino)-N 1 -(4-Methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0159]

[0160] The synthesis method was the same as that for intermediate F-1, yielding 48 mg of off-white solid, with a yield of 44%. 1H NMR (400MHz, DMSO-d6) δ10.96 (s, 1H, NH), 8.60 (d, J=5.1Hz, 1H, Ar-H), 8.45-8.35 (m, 1H, NH), 8.16 (d, J=8.3Hz, 2H , Ar-H), 7.95 (d, J=8.4Hz, 2H, Ar-H), 7.54 (s, 1H, Ar-H), 7.44-7.40 (m, 1H, NH), 7.40-7.35 (m, 1H, Ar-H), 7.05-6.9 2 (m, 1H, NH), 4.38-4.20 (m, 1H, CH), 3.91-3.87 (m, 1H, CH), 3.87 (s, 3H, CH3), 3.67-3.55 (m, 1H, CH), 3.12-3.03 (m, 1H, CH), 2.40 (s, 3H, CH3), 2.09-1.94 (m, 1H, CH), 1.90-1.72 (m, 1H, CH), 1.61 (s, 6H, CH3×2); ESI-MS: m / z=655[M+H] + .

[0161] Synthesis of intermediate F-7

[0162] (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(4-Methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0163]

[0164] The synthesis method was the same as that for intermediate F-1, yielding 55 mg of off-white solid, with a yield of 48%. 1H NMR (400MHz, DMSO-d6) δ11.05 (s, 1H, NH), 8.60 (d, J=4.9Hz, 1H, Ar-H), 7.98 (d, J=7.6Hz, 2H, Ar-H), 7.70-7 .60 (m, 1H, NH), 7.58-7.52 (m, 3H, Ar-H), 7.50-7.44 (m, 1H, NH), 7.44-7.38 (m, 1H, Ar-H), 7.07-6.96 (m, 1H, N H), 4.50 (s, 2H, CH2), 4.41-4.27 (m, 1H, CH), 4.08-3.95 (m, 1H, CH), 3.86 (s, 3H, CH3), 3.63-3.57 (m, 1H, CH) , 3.46-3.38 (m, 1H, CH), 2.41 (s, 3H, CH3), 2.15-1.98 (m, 2H, CH), 1.61 (s, 6H, CH3×2); ESI-MS: m / z=669[M+H] + .

[0165] Synthesis of intermediate F-8

[0166] (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-(tert-butyl)pyridin-4-yl)-4-methylthiazolyl-2-yl)pyrrolidine-1,2-dicarboxamide

[0167]

[0168] The synthesis method was the same as that used for intermediate F-1, yielding 50 mg of a white solid, yield: 47%. ESI-MS: m / z = 615 [M+H] + .

[0169] Synthesis of intermediate F-9

[0170] (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-Cyclobutylpyridin-4-yl)-4-methylthiazolyl-2-yl)-1,2-dicarboxylamide

[0171]

[0172] The synthesis method was the same as that used for intermediate F-1, yielding 45 mg of a white solid, yield: 45%. ES1-MS: m / z = 613 [M+H] + .

[0173] Synthesis of intermediate F-10

[0174] (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(4-Methyl-5-(2-(1-methylcyclopropyl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0175]

[0176] The synthesis method was the same as that used for intermediate F-1, yielding 37 mg of a white solid, yield: 50%. ESI-MS: m / z = 613 [M+H] + .

[0177] Synthesis of intermediate F-11

[0178] (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-(1-cyanocyclopropyl)pyridin-4-yl)-4-methylthiazo-2-yl)-1,2-dicarboxylamide

[0179]

[0180] The synthesis method was the same as that used for intermediate F-1, yielding 48 mg of a white solid, with a yield of 49%. ESI-MS: m / z = 624 [M+H] + .

[0181] Synthesis of intermediate F-12

[0182] (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(4-Methyl-5-(2-(1-(trifluoromethyl)cyclopropyl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0183]

[0184] The synthesis method was the same as that used for intermediate F-1, yielding 36 mg of a white solid, with a yield of 48%. ESI-MS: m / z = 667 [M+H] + .

[0185] Synthesis of intermediate F-13

[0186] (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(4-Methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyrimidin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0187]

[0188] The synthesis method was the same as that used for intermediate F-1, yielding 42 mg of a white solid, yield: 53%. ESI-MS: m / z = 670 [M+H] + .

[0189] Synthesis of intermediate F-14

[0190] (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-(tert-butyl)pyrimidin-4-yl)-4-methylthiazolyl-2-yl)pyrrolidine-1,2-dicarboxamide

[0191]

[0192] The synthesis method was the same as that used for intermediate F-1, yielding 38 mg of a white solid, yield: 49%. ESI-MS: m / z = 616 [M+H] + .

[0193] Synthesis of intermediate F-15

[0194] (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-Cyclopropylpyrimidin-4-yl)-4-methylthiazolyl-2-yl)pyrrolidine-1,2-dicarboxamide

[0195]

[0196] The synthesis method was the same as that used for intermediate F-1, yielding 40 mg of a white solid, with a yield of 39%. ESI-MS: m / z = 600 [M+H] + .

[0197] Synthesis of intermediate F-16

[0198] (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(4-Methyl-5-(2-(1-methylcyclopropyl)pyrimidin-4-yl)thiazolyl-2-yl)pyrrolidine-1,2-dicarboxylamide

[0199]

[0200] The synthesis method was the same as that used for intermediate F-1, yielding 42 mg of a white solid, with a yield of 41%. ESI-MS: m / z = 614 [M+H] + .

[0201] Synthesis of intermediate F-17

[0202] (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-(azacyclobutan-1-yl)pyrimidin-4-yl)-4-methylthiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0203]

[0204] The synthesis method was the same as that used for intermediate F-1, yielding 51 mg of a white solid, yield: 42%. ESI-MS: m / z = 615 [M+H] + .

[0205] Synthesis of intermediate F-18

[0206] (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-(diethylamino)pyrimidin-4-yl)-4-methylthiazolyl-2-yl)pyrrolidine-1,2-dicarboxylamide

[0207]

[0208] The synthesis method was the same as that used for intermediate F-1, yielding 55 mg of a white solid, yield: 56%. ESI-MS: m / z = 631 [M+H] + .

[0209] Synthesis of intermediate F-19

[0210] (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(4-chloro-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0211]

[0212] The synthesis method was the same as that used for intermediate F-1, yielding 43 mg of a white solid, yield: 46%. ESI-MS: m / z = 689 [M+H] + .

[0213] Synthesis of 4-substituted-L-prolyl PI3Kα / HDAC6 dual inhibitors (Examples 1-22)

[0214]

[0215] Intermediate F, containing an ester group, reacts with hydroxylamine under alkaline conditions to generate a 4-substituted-L-prolyl PI3Kα / HDAC6 dual inhibitor containing an isohydroxamic acid group. The reaction uses sodium hydroxide (NaOH) as the alkali, tetrahydrofuran / methanol (THF / MeOH) as the solvent, reacts at room temperature, and lasts for 1 hour.

[0216] Examples of the synthesized end products are given below. These examples, along with the examples above, are for illustrative purposes only and should not be construed as limiting the scope of the invention. Furthermore, it should be understood that after reading this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined by the appended claims.

[0217] Example 1

[0218] (2S,4R)-4-(4-(hydroxycarbamoyl)benzoyl)-N 1 -(4-Methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0219]

[0220] The intermediate methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate (49 mg, 0.08 mmol) was dissolved in an equal volume of tetrahydrofuran and methanol. 1 mL of methanol was added for every 0.1 mmol of substrate. NaOH (26 mg, 0.64 mmol) and NH₂OH (50 wt.% in water) were added under ice-bath stirring, with 1 mL of NH₂OH added for every 1 mmol of substrate. The reaction was carried out at room temperature for one hour. After the reaction, the pH of the mixture was adjusted to 7 to obtain the crude product. The crude product was purified by thin-layer chromatography (25% MeOH / CH₂Cl₂) to give a white solid (25 mg, 0.04 mmol), yield: 40%. 1H NMR (400MHz, DMSO-d6) δ11.09 (s, 1H, NH), 10.77 (s, 1H, NH), 8.91 (s, 1H, OH), 8.59 (d, J=4.6Hz, 1H, Ar-H), 8. 31-8.19 (m, 1H, NH), 7.94 (d, J=7.4Hz, 2H, Ar-H), 7.85 (d, J=7.8Hz, 2H, Ar-H), 7.53 (d, J=8.2Hz, 2H, Ar-H), 7. 43-7.38 (m, 1H, NH), 7.10-6.97 (m, 1H, NH), 4.66-4.55 (m, 1H, CH), 4.46-4.31 (m, 1H, CH), 3.94-3.80 (m, 1H, CH) ), 3.58-3.49 (m, 1H, CH), 2.40 (s, 3H, CH3), 2.35-2.23 (m, 1H, CH), 2.22-2.04 (m, 1H, CH), 1.61 (s, 6H, CH3×2); 13 CNMR (150MHz, DMSO-d6) δ174.02, 171.18, 166.22, 158.99, 149.57, 146.09, 145.50, 141.41, 132.6 3, 130.12, 129.73, 129.09, 127.95, 127.23, 123.63, 121.75, 120.72, 117.70, 51.47, 46.80 (q, JC- F =24.0Hz), 31.62, 30.30, 29.48, 28.32, 22.55, 21.92, 14.42; ESI-HRMS: m / zcalcd for C 27 H 28 F3N7O5S[M+H]+620.1898; found 620.1900.

[0221] Example 2

[0222] (2S,4R)-4-(2-(4-(hydroxycarbamoyl)phenyl)acetamido)-N 1 -(4-Methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0223]

[0224] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with methyl 4-(2-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl)amino)-2-oxoethyl)benzoate, yielding 17 mg of off-white solid, yield: 50%. 1 H NMR (400MHz, DMSO-d6) δ10.43 (s, 1H, NH), 9.12-8.94 (m, 1H, NH), 8.66 (s, 1H, OH), 8.56 (d, J=4.2Hz, 1H, Ar-H), 7. 92-7.74 (m, 1H, NH), 7.65 (d, J=7.7Hz, 2H, Ar-H), 7.52 (s, 1H, Ar-H), 7.49-7.44 (m, 1H, NH), 7.38 (d, J=2.6Hz, 1H, A r-H), 7.27 (d, J=7.5Hz, 2H, Ar-H), 7.06-6.93 (m, 1H, NH), 4.38-4.23 (m, 2H, CH), 4.19-4.07 (m, 1H, CH), 3.78-3.70 (m, 1H, CH), 3.44 (s, 2H, CH2), 2.37 (s, 3H, CH3), 2.11-2.04 (m, 1H, CH), 1.99-1.90 (m, 1H, CH), 1.58 (s, 6H, CH3×2); 13 C NMR (150MHz, DMSO-d6) δ174.00, 172.57, 171.86, 164.96, 160.02, 158.96, 150.46, 149.56, 147. 46, 144.59, 141.83, 141.43, 132.36, 129.36, 127.86, 127.24, 121.76, 120.72, 51.91, 46.78 (q, J C-F =24.0Hz), 42.34, 31.62, 30.29, 29.49, 29.17, 22.57, 21.91, 14.43; ESI-HRMS: m / z calcd for C 28 H 30 F3N7O5S[M+H]+634.2054; found 634.2059.

[0225] Example 3

[0226] (2S,4R)-4-(5-(hydroxycarbamoyl)pyridine-2-acylamino)-N 1 -(4-Methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0227]

[0228] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with (2S,4R)-4-(5-(methoxycarbonyl)pyridin-2-acylamino)-N 1 -(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)pyrrolidine-1,2-dicarboxamide, to give 14 mg of yellow solid, yield: 30%. 1 H NMR (400MHz, DMSO-d6) δ11.40-10.96 (m, 2H, NH), 9.52-9.30 (m, 1H, NH), 9.16 (s, 1H, OH), 9.00 (s, 1H, Ar-H), 8.60 (d, J=4.5Hz, 1H, Ar-H), 8.40-8.28 (m, 1H, Ar-H), 8.15-8.06 (m, 1H, Ar-H), 7.59-7.50 (m, 2H, Ar-H), 7.4 5-7.37 (m, 1H, NH), 7.11-7.00 (m, 1H, NH), 4.78-4.63 (m, 1H, CH), 4.45-4.35 (m, 1H, CH), 3.95-3.86 (m, 1H, CH) ), 3.61-3.48 (m, 1H, CH), 2.40 (s, 3H, CH3), 2.38-2.30 (m, 1H, CH), 2.12-2.00 (m, 1H, CH), 1.61 (s, 6H, CH3×2); 13 C NMR (150MHz, DMSO-d6) δ174.04, 173.98, 169.09, 163.92, 158.98, 151.85, 149.58, 147.35, 141. 37, 136.99, 131.19, 130.12, 128.37, 127.85, 122.40, 121.78, 120.75, 55.42, 49.03, 46.79 (q, JC-F =24.0Hz), 31.62, 30.29, 29.48, 22.56, 21.91, 14.43; ESI-HRMS: m / z calcd for C 26 H 27 F3N8O5S[M+H]+621.1850; found621.1849.

[0229] Example 4

[0230] (2S,4R)-4-((5-(hydroxycarbamoyl)pyridin-2-yl)amino)-N 1 -(4-Methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0231]

[0232] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with methyl 2-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl)amino)pyridine-5-carboxylate, yielding 15 mg of yellow solid, with a yield of 43%. 1 H NMR (400MHz, DMSO-d6) δ11.23 (s, 1H, NH), 8.57 (s, 1H, OH), 8.55 (s, 1H, Ar-H), 7.78 (d, J=8.5Hz, 1H, Ar-H), 7.68- 7.61 (m, 1H, NH), 7.52-7.44 (m, 2H, Ar-H), 7.39-7.36 (m, 1H, Ar-H), 7.35-7.28 (m, 1H, NH), 7.25-7.17 (m, 1H, NH), 7.04-6.93 (m, 1H, NH), 6.50 (d, J=8.9Hz, 1H, Ar-H), 4.60-4.54 (m, 1H, CH), 4.41-4.32 (m, 1H, CH), 4.21-4.12 (m, 1 H, CH), 3.90-3.84 (m, 1H, CH), 2.36 (s, 3H, CH3), 2.15-2.12 (m, 1H, CH), 1.98-1.93 (m, 1H, CH), 1.57 (s, 6H, CH3×2);13 C NMR (100MHz, CDCl3-d6) δ173.45, 166.29, 159.95, 159.43, 151.04, 148.97, 141.43, 140.47, 1 38.81, 138.16, 132.82, 129.56, 126.75, 121.16, 120.69, 115.46, 59.01, 51.75, 46.85 (q, JC- F =24.0Hz), 35.43, 29.68, 21.89, 21.87, 15.43, 12.04; ESI-HRMS: m / z calcdfor C 25 H 27 F3N8O4S[M+H]+593.1901; found 593.1909.

[0233] Example 5

[0234] (2S,4R)-4-((5-(hydroxycarbamoyl)pyrimidin-2-yl)amino)-N 1 -(4-Methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0235]

[0236] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with methyl 2-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl)amino)pyrimidine-5-carboxylate, yielding 36 mg of yellow solid, with a yield of 67%. 1H NMR (600MHz, DMSO-d6) δ11.90 (s, 1H, NH), 11.09 (s, 1H, OH), 8.82-8.75 (m, 1H, Ar-H), 8.74-8.69 (m, 1H, Ar-H), 8. 69-8.61 (m, 1H, NH), 8.60 (d, J=5.0Hz, 1H, Ar-H), 8.42-8.28 (m, 1H, NH), 7.55 (s, 1H, Ar-H), 7.51-7.46 (m, 1H, NH), 7.41 (d, J=4.2Hz, 1H, Ar-H), 7.09-6.97 (m, 1H, NH), 4.72-4.58 (m, 1H, CH), 4.44-4.30 (m, 1H, CH), 3.97-3.83 (m, 1 H, CH), 3.58-3.48 (m, 1H, CH), 2.40 (s, 3H, CH3), 2.26-2.15 (m, 1H, CH), 2.13-2.01 (m, 1H, CH), 1.61 (s, 6H, CH3×2); 13 C NMR (150MHz, DMSO-d6) δ174.02, 173.99, 166.29, 163.22, 162.82, 160.24, 158.98, 149. 57, 141.37, 131.60, 129.72, 127.85, 125.97, 121.76, 120.74, 59.27, 51.63, 46.79 (q, J C-F =24.0Hz), 35.43, 31.76, 29.50, 22.57, 21.90, 16.62; ESI-HRMS: m / z calcd for C 24 H 26 F3N9O4S[M+H]+594.1854; found 594.1851.

[0237] Example 6

[0238] (2S,4R)-4-((4-(hydroxycarbamoyl)phenyl)sulfonylamino)-N 1 -(4-Methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0239]

[0240] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with (2S,4R)-4-((4-(methoxycarbonyl)phenyl)sulfonylamino)-N 1 -(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)pyrrolidine-1,2-dicarboxamide, to give 18 mg of yellow solid, yield: 43%. 1 H NMR (400MHz, DMSO-d6) δ11.36 (s, 1H, NH), 10.09 (s, 1H, OH), 9.34-9.12 (m, 1H, NH), 8.59 (d, J=4.3Hz, 1H, Ar-H), 8. 53-8.36 (m, 1H, NH), 7.97 (d, J=7.6Hz, 2H, Ar-H), 7.89 (d, J=7.9Hz, 2H, Ar-H), 7.54 (s, 1H, Ar-H), 7.49-7.43 (m, 1H , NH), 7.42-7.36 (m, 1H, Ar-H), 7.06-6.93 (m, 1H, NH), 4.35-4.17 (m, 1H, CH), 3.91-3.77 (m, 1H, CH), 3.76-3.61 (m, 1H, CH), 3.07-2.96 (m, 1H, CH), 2.39 (s, 3H, CH3), 2.12-1.95 (m, 1H, CH), 1.89-1.73 (m, 1H, CH), 1.60 (s, 6H, CH3×2); 13 C NMR (150MHz, DMSO-d6) δ173.80, 169.24, 160.14, 158.97, 149.58, 147.90, 145.89, 141.35, 135.9 1, 131.08, 130.12, 129.60, 127.85, 126.24, 124.71, 121.78, 120.76, 58.43, 52.30, 46.78 (q, JC- F =23.0Hz), 31.62, 30.29, 29.48, 22.56, 21.90, 14.43; ESI-HRMS: m / z calcdfor C 26 H 28 F3N7O6S2[M+H]+656.1568; found656.1579.

[0241] Example 7

[0242] (2S,4R)-4-(((4-(hydroxycarbamoyl)phenyl)methyl)sulfonylamino)-N 1 -(4-Methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0243]

[0244] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)pyrrolidine-1,2-dicarboxamide, to give 17 mg of yellow solid, yield: 25%. 1 H NMR (400MHz, DMSO-d6) δ11.16 (s, 1H, NH), 8.59 (s, 1H, OH), 7.94 (d, J=6.6Hz, 1H, Ar-H), 7.93-7.84 (m, 1H, NH), 7.82-7.69 (m, 1H, NH), 7.58-7.49 (m, 2H, Ar-H), 7.47-7.31 (m, 4H, Ar-H), 7.20-7.14 (m, 1H, NH), 7.10 -6.93(m,1H,NH),4.44(s,2H,CH2),4.38-4.26(m,1H,CH),4.17-4.09(m,1H,CH),4.09-3.96(m,1H,CH),3 .95-3.76 (m, 1H, CH), 2.41 (s, 3H, CH3), 2.21-2.11 (m, 1H, CH), 2.01-1.91 (m, 1H, CH), 1.61 (s, 6H, CH3×2); 13 C NMR (150MHz, DMSO-d6) δ173.76, 170.35, 163.24, 158.97, 149.57, 143.57, 141.33, 136.83, 1 32.02, 131.60, 129.72, 129.14, 128.43, 127.84, 126.96, 121.77, 120.74, 65.50, 46.78 (q, J C-F=24.0Hz), 45.72, 31.62, 30.28, 29.48, 22.56, 21.90, 19.12, 8.99; ESI-HRMS: m / z calcd for C 27 H 30 F3N7O6S2[M+H]+670.1724; found 670.1736.

[0245] Example 8

[0246] (2S,4R)-4-(((4-(hydroxycarbamoyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-(tert-butyl)pyridin-4-yl)-4-methylthiazolyl-2-yl)pyrrolidine-1,2-dicarboxamide

[0247]

[0248] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-(tert-butyl)pyridin-4-yl)-4-methylthiazolyl-2-yl)pyrrolidine-1,2-dicarboxamide, to give 20 mg of white solid, yield: 36%. ESI-MS: m / z = 616 [M+H] + .

[0249] Example 9

[0250] (2S,4R)-4-(((4-(hydroxycarbamoyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-cyclobutylpyridin-4-yl)-4-methylthiazolyl-2-yl)pyrrolidine-1,2-dicarboxylamide

[0251]

[0252] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N1 -(5-(2-cyclobutylpyridin-4-yl)-4-methylthiazolyl-2-yl)-1,2-dicarboxamide, to give 30 mg of white solid, yield: 41%. ESI-MS: m / z = 614 [M+H] + .

[0253] Example 10

[0254] (2S,4R)-4-(((4-(hydroxycarbamoyl)phenyl)methyl)sulfonylamino)-N 1 -(4-Methyl-5-(2-(1-methylcyclopropyl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0255]

[0256] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(4-methyl-5-(2-(1-methylcyclopropyl)pyridin-4-yl)thiazolyl)pyrrolidine-1,2-dicarboxamide, to give 29 mg of white solid, yield: 39%. ESI-MS: m / z = 614 [M+H] + .

[0257] Example 11

[0258] (2S,4R)-4-(((4-(hydroxycarbamoyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-(1-cyanocyclopropyl)pyridin-4-yl)-4-methylthiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0259]

[0260] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1-(5-(2-(1-cyanocyclopropyl)pyridin-4-yl)-4-methylthiazolyl-2-yl)-1,2-dicarboxamide, to give 29 mg of white solid, yield: 39%. ESI-MS: m / z = 625 [M+H] + .

[0261] Example 12

[0262] (2S,4R)-4-(((4-(hydroxycarbamoyl)phenyl)methyl)sulfonylamino)-N 1 -(4-Methyl-5-(2-(1-(trifluoromethyl)cyclopropyl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0263]

[0264] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(4-methyl-5-(2-(1-(trifluoromethyl)cyclopropyl)pyridin-4-yl)thiazolyl)pyrrolidine-1,2-dicarboxamide, to give 21 mg of white solid, yield: 29%. ESI-MS: m / z = 668 [M+H] + .

[0265] Example 13

[0266] (2S,4R)-4-(((4-(hydroxycarbamoyl)phenyl)methyl)sulfonylamino)-N 1 -(4-Methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyrimidin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0267]

[0268] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1-(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyrimidin-4-yl)thiazolyl)pyrrolidine-1,2-dicarboxamide, to give 26 mg of white solid, yield: 31%. ESI-MS: m / z = 671 [M+H] + .

[0269] Example 14

[0270] (2S,4R)-4-(((4-(hydroxycarbamoyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-(tert-butyl)pyrimidin-4-yl)-4-methylthiazolyl-2-yl)-4-(cyclopropanesulfonylamino)pyrrolidine-1,2-dicarboxylamino

[0271]

[0272] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-(tert-butyl)pyrimidin-4-yl)-4-methylthiazolyl-2-yl)pyrrolidine-1,2-dicarboxamide, to give 32 mg of white solid, yield: 26%. ESI-MS: m / z = 617 [M+H] + .

[0273] Example 15

[0274] a)(2S,4R)-4-(((4-(hydroxycarbamoyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-Cyclopropylpyrimidin-4-yl)-4-methylthiazolyl-2-yl)pyrrolidine-1,2-dicarboxamide

[0275]

[0276] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1-(5-(2-cyclopropylpyrimidin-4-yl)-4-methylthiazolyl-2-yl)pyrrolidine-1,2-dicarboxamide, to give 39 mg of white solid, yield: 32%. ESI-MS: m / z = 601 [M+H] + .

[0277] Example 16

[0278] (2S,4R)-4-(((4-(hydroxycarbamoyl)phenyl)methyl)sulfonylamino)-N 1 -(4-Methyl-5-(2-(1-methylcyclopropyl)pyrimidin-4-yl)thiazolyl-2-yl)pyrrolidine-1,2-dicarboxylamide

[0279]

[0280] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(4-methyl-5-(2-(1-methylcyclopropyl)pyrimidin-4-yl)thiazolyl-2-yl)pyrrolidine-1,2-dicarboxamide, to give 35 mg of white solid, yield: 41%. ESI-MS: m / z = 615 [M+H] + .

[0281] Example 17

[0282] (2S,4R)-4-(((4-(hydroxycarbamoyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-(azacyclobutan-1-yl)pyrimidin-4-yl)-4-methylthiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0283]

[0284] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1-(5-(2-(azacyclobutan-1-yl)pyrimidin-4-yl)-4-methylthiazo-2-yl)pyrrolidine-1,2-dicarboxamide, to give 37 mg of white solid, yield: 39%. ESI-MS: m / z = 616 [M+H] + .

[0285] Example 18

[0286] (2S,4R)-4-(((4-(hydroxycarbamoyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-(diethylamino)pyrimidin-4-yl)-4-methylthiazolyl-2-yl)pyrrolidine-1,2-dicarboxylamide

[0287]

[0288] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1 -(5-(2-(diethylamino)pyrimidin-4-yl)-4-methylthiazolyl-2-yl)pyrrolidine-1,2-dicarboxamide, to give 39 mg of white solid, yield: 45%. ESI-MS: m / z = 632 [M+H] + .

[0289] Example 19

[0290] (2S,4R)-4-(((4-(hydroxycarbamoyl)phenyl)methyl)sulfonylamino)-N 1 -(4-chloro-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazo-2-yl)pyrrolidine-1,2-dicarboxylamide

[0291]

[0292] This embodiment uses the same implementation method as Example 1, except that methyl 4-(((3R,5S)-5-carbamoyl-1-((4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)carbamoyl)pyrrolidine-3-yl))carbamoyl)benzoate is replaced with (2S,4R)-4-(((4-(methoxycarbonyl)phenyl)methyl)sulfonylamino)-N 1-(4-chloro-5-(2-(1,1,1-trifluoro-2-methylpropane-2-yl)pyridin-4-yl)thiazolyl)pyrrolidine-1,2-dicarboxamide, to give 26 mg of white solid, yield: 34%. ESI-MS: m / z = 691 [M+H] + .

[0293] Test Example 1: Inhibitory activity of the compounds of this invention against PI3Kα and HDAC6

[0294] The in vitro PI3Kα inhibitory activity of the compounds of this invention was determined using the Kinase-Glo Plus Luminescent Kinase assay. First, the test compounds were diluted to the desired concentration, and 2.5 μL of each compound was added to a 384-well plate. A 1x kinase buffer was prepared using 50 mM HEPES (pH 7.5), 3 mM MgCl2, 1 mM EGTA, 100 mM NaCl, 0.03% CHAPS, and 2 mM DTT. PI3Kα was diluted to a concentration of 1.65 nM using the kinase buffer, and 2.5 μL of this kinase solution was also added to a 384-well plate. The substrates PIP2 and ATP were diluted to concentrations of 50 μM and 25 μM respectively using the kinase buffer, and 5 μL of the substrate solution was added to the 384-well plate. After reacting at room temperature for 1 hour, 10 μL of Kinase-Glo reagent was added to the reaction wells of the assay plate to terminate the reaction. Centrifuge the resulting mixture, then gently shake on a shaker for 15 minutes and measure its RLM value using a microplate reader. The inhibition rate is calculated as follows: Inhibition rate (%) = (sample RLM - min) / (max - min) × 100, where "min" represents the RLM of the enzyme-free control wells and "max" represents the RLM of the wells containing the DMSO control. IC50 50 The values ​​were calculated using Graphpad 5.0 software.

[0295] The in vitro HDAC6 enzyme inhibitory activity of the compounds of this invention was determined by the following steps. First, the test compound was diluted to the desired concentration, and the kinase was diluted to the desired concentration using 1x kinase buffer. Trypsin and Ac peptide substrate were added to 1x assay buffer to prepare a substrate solution. 15 μL of the enzyme solution was transferred to an assay plate and incubated at room temperature for 15 minutes. 10 μL of substrate solution was added to each well to begin the reaction. Paradigm measurements were performed at an excitation wavelength of 355 nm and an emission wavelength of 460 nm. The inhibition rate was calculated as follows: Inhibition rate (%) = (Max - Signal) / (Max - Min) * 100, Y = Bottom + (Top - Bottom) / (1 + (IC) * 100) * 100. 50 / X)*HillSlope; Y is the inhibition percentage, and X is the compound concentration.

[0296] Table 1. Inhibitory activity of the invented compounds against P13Kα

[0297]

[0298]

[0299] "++++" represents 1-5nM; "+++" represents 5-10nM; "++" represents 10-50nM; "+" represents >50nM.

[0300] As shown in Table 1, most of the compounds in this invention exhibit significant dual PI3Kα / HDAC6 inhibitory activity. Among these compounds with significant dual PI3Kα / HDAC6 inhibitory activity, most compounds show inhibitory activity against PI3Kα that is superior to or comparable to Alpelisib, while their inhibitory activity against HDAC6 is comparable to SAHA.

[0301] Test Example 2: Selectivity test of the compound of the present invention for PI3Kα

[0302] The in vitro enzyme inhibitory activities of the compounds in this invention were determined using the ADP-Glo ​​Kinase assay. Alpelisib, a commercially available small molecule inhibitor of PI3Kα, was selected as a positive control. First, the test compounds were diluted to the desired concentrations, and 2.5 μL of each compound was added to a 384-well plate. A 1x kinase buffer was prepared using 50 mM HEPES (pH 7.5), 3 mM MgCl2, 1 mM MEGTA, 100 mM NaCl, 0.03% CHAPS, and 2 mM DTT. PI3Kα was diluted to a concentration of 1.65 nM using the kinase buffer, and PI3Kβ, PI3Kγ, and PI3Kδ were diluted to 4.8 nM, 7.6 nM, and 5.7 nM, respectively. 2.5 μL of this kinase solution was also added to a 384-well plate. The substrates PIP2 and ATP were diluted to concentrations of 50 μM and 25 μM using the kinase buffer, and 5 μL of each substrate solution was added to the 384-well plate. First, transfer 5 μL of the reaction mixture to a new 384-well plate, then add 5 μL of ADP-Glo ​​reagent to terminate the reaction. Centrifuge the mixture, vortex for 40 minutes, then add 10 μL of kinase assay reagent to each well, continue vortexing for 1 hour, and finally measure the RLM value using a microplate reader. The inhibition rate is calculated as follows: Inhibition rate (%) = (sample RLM - min) / (max - min) × 100, where "min" represents the RLM of the enzyme-free control well, and "max" represents the RLM of the well containing DMSO control. IC50 50 The values ​​were calculated using Graphpad 5.0 software.

[0303] The assay of PI3Kα, PI3Kβ, PI3Kγ, and PI3Kδ enzyme inhibitory activities reflects the subtype-selective inhibition of PI3Kα by compounds. The following data, based on the inhibitory activity data of compounds exhibiting significant PI3Kα and HDAC6 inhibitory activities, further illustrate their selectivity for PI3Kα. It should not be construed that only the following compounds possess selective PI3Kα inhibitory activity.

[0304] Table 2. Selectivity test of compounds for PI3Kα

[0305]

[0306] "++++" represents 100-500; "+++" represents 50-100; "++" represents 10-50; "+" represents 5-10.

[0307] As shown in Table 2, Examples 3, 7, 12, 13, and 16, which conform to general formula (I), have excellent subtype selectivity for PI3Kα. Their selectivity is better than or comparable to that of Alpelisib, indicating that Examples 3, 7, 12, 13, and 16 in this invention significantly inhibit PI3Kα subtypes while reducing the toxicity caused by inhibition of other PI3K subtypes.

[0308] Test Example 3: Selectivity test of the compound of the present invention for HDAC6

[0309] The assays of HDAC1, HDAC2, HDAC3, HDAC6, HDAC8, HDAC10, and HDAC11 enzyme inhibitory activities reflect the selective inhibition of HDAC6 subtypes by the compounds. The following data, using the inhibitory activity data of compounds exhibiting significant PI3Kα and HDAC6 inhibitory activities against HDAC1, 2, 3, 6, 8, 10, and 11, further illustrate their selectivity for HDAC6. It should not be construed that only the following compounds exhibit selective HDAC6 inhibition. The assay methods for the inhibitory activities of compounds against other HDAC subtypes are the same as those for HDAC1 inhibitory activity, only the substrate in the catalytic reaction system is changed when testing the corresponding enzyme inhibitory activity.

[0310] Table 3. Selectivity test of compounds for HDAC6

[0311]

[0312] "++++" represents 100-500; "+++" represents 50-100; "++" represents 10-50; "+" represents 5-10.

[0313] Table 3 shows that the broad-spectrum HDAC inhibitor SAHA exhibits relatively similar activity against various HDAC isoforms, lacking selectivity for the HDAC6 isoform. In contrast, Examples 3, 7, 12, 13, and 16, conforming to general formula (I), demonstrate excellent isoform selectivity for HDAC6. This indicates that Examples 3, 7, 12, 13, and 16 of this invention significantly inhibit the HDAC6 isoform while also reducing the toxicity caused by inhibition of other HDAC isoforms.

[0314] The above results demonstrate that the compounds of this invention can simultaneously and selectively inhibit PI3Kα and HDAC6, which are closely related to tumor development and progression. This is highly beneficial in reducing the toxicity caused by inhibiting other PI3K and HDAC subtypes with normal physiological functions. Therefore, the compounds of this invention represent a novel class of antitumor compounds with promising applications.

[0315] The technical features of the embodiments described above can be combined arbitrarily. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as the combination of these technical features does not contradict each other, it should be considered within the scope of this specification. The embodiments described above only illustrate several implementations of the present invention, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make several modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A PI3Ka / HDAC6 isoform selective dual inhibitor which is a compound having the general formula (I) as follows: ###0001### wherein, X, Y, Z are all CH; or Y, Z are CH and X is N; R2 is selected from methyl or chloro; R1is selected from 2. A PI3Ka / HDAC6 isoform selective dual inhibitor which is selected from the following compounds: ###0002### ###0003### ###0004### ###0005### ###0006### ###0007### ###0008### ###0009### ###0010### ###0011### ###0012### ###0013### ###0014### ###0015### ###0016### ###0017### ###0018### ###0019### ###0020### ###0021### ###0022### ###0023### ###0024### ###0025### ###0026### ###0027### ###0028### ###0029### ###0030### ###0031### L is Ring A is C 6-14 aryl, C 5-14 heteroaryl, R3is independently selected from hydrogen, halogen, hydroxy, cyano, C 1-6 alkyl, or C 1-6 alkoxy. ​

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