Oxadiazole histone deacetylase / GSK-3beta dual inhibitor and application

Abstract

The invention discloses an oxadiazole histone deacetylase / GSK-3beta dual inhibitor and application, and relates to the technical field of medical chemistry. The invention provides a compound or a pharmaceutically acceptable salt thereof, and provides a structural general formula of the compound. A pharmaceutical composition containing the compound is applied to preparation of drugs for preventing and / or treating histone deacetylase / GSK-3beta diseases. The compound has a synergistic anti-tumor effect, namely GSK inhibition (such as GSK-3beta); signal channels of Wnt / beta-catenin, NF-kB and the like are regulated and controlled; tumor cell proliferation is inhibited; apoptosis is induced; the chemotherapy / radiotherapy sensitivity is enhanced, and the drug resistance is overcome; hDAC inhibition: activating cancer suppressor genes through epigenetic regulation (histone acetylation), inhibiting tumor growth and inducing cell cycle arrest and differentiation; the double targets act at the same time, so that complementary pathways can be blocked, the risk of single drug resistance is reduced, and the anti-tumor effect is enhanced.

Description

Technical Field

[0001] The present invention relates to the technical field of medicinal chemistry, and in particular to an oxadiazole histone deacetylase / GSK-3β dual inhibitor and its application. Background Art

[0002] Orderly gene transcriptional regulation is essential for maintaining normal cellular function. Disruption of gene transcriptional regulation can lead to cancer. Acetylation and deacetylation of core histones are closely linked to gene regulation. Histone acetylation and deacetylation are mediated by a pair of antagonistic proteases: histone acetyltransferases (HATs) and histone deacetylases (HDACs). HDACs are a group of enzymes that regulate a range of biological effects at the chromatin level by inducing histone deacetylation, including chromatin reorganization, transcriptional activation or repression, cell cycle progression, cell differentiation, and apoptosis. HDACs are particularly involved in regulating gene transcriptional expression following cell activation. Histone deacetylase inhibitors, by inhibiting HDAC activity, induce apoptosis and differentiation, and inhibit cell proliferation, and are considered promising anticancer drug targets. Currently, research on HDAC inhibitors spans a wide range of tumor types, including hematologic malignancies, melanoma, breast cancer, ovarian cancer, prostate cancer, lung cancer, and colon cancer. GSK3β (glycogen synthase kinase 3β, GSK3β) is a ubiquitously expressed serine / threonine kinase involved in regulating cell proliferation, differentiation, metabolism, and apoptosis. Recent studies have revealed that GSK3β plays a crucial role in tumorigenesis, progression, and treatment resistance, with its specific functions highly dependent on tumor type, microenvironment, and signaling pathway context.

[0003] HDAC inhibitors and GSK3β (glycogen synthase kinase 3β) inhibitors each have unique mechanisms of action in tumor treatment. Studies have shown that their combined use may produce a synergistic anti-tumor effect and exhibit significant advantages in overcoming drug resistance and enhancing apoptosis induction. Based on this, the present invention has developed a novel class of HDAC / GSK3β dual inhibitors, hoping to produce new, selectable HDAC / GSK3β dual inhibitors for the prevention and / or treatment of diseases associated with uncontrolled histone deacetylase or GSK3β activity, particularly tumors. Summary of the Invention

[0004] In order to solve the above technical problems, the purpose of the present invention is to provide an oxadiazole histone deacetylase GSK-3β dual inhibitor and its application, which is used to comprehensively reverse the malignant phenotype of tumors. The simultaneous action of dual targets can block complementary pathways, reduce the risk of single drug resistance, and enhance the anti-tumor effect.

[0005] The present invention solves the above technical problems by providing a compound or a pharmaceutically acceptable salt thereof, wherein the compound has the general structural formula:

[0006]

[0007] Among them, R 1 、R 2 are independently selected from hydrogen, alkyl, alkenyl, or alkynyl;

[0008] Q 1 is aryl or Het; aryl or Het is optionally replaced by at least one R 3 Replacement, R 3 is hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl or Het;

[0009] Q 2 is aryl or Het; aryl or Het is optionally replaced by at least one R 4 Replacement, R 4 is hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylamino, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl or Het;

[0010] The alkyl group is a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms connected to a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms;

[0011] Alkoxy is a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms connected to a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms; wherein each carbon atom is optionally substituted by oxygen;

[0012] Alkylamino is a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms connected to a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms; wherein each carbon atom is optionally substituted by an NH group;

[0013] Alkoxyalkyl is an alkoxy group connected to an alkyl group;

[0014] Alkenyl or alkynyl is a straight or branched unsaturated hydrocarbon group having 1 to 6 carbon atoms and containing double or triple bonds;

[0015] Aryl is a carbocyclic ring of phenyl, naphthyl, acenaphthyl or tetrahydronaphthyl, optionally substituted by 1, 2 or 3 substituents, wherein the substituents are hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl or Het;

[0016] Aralkyl or diarylalkyl is an aryl group connected to an alkyl group;

[0017] Het is a monocyclic heterocycle selected from pyrrolyl, pyrazolyl, imidazolyl, furyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl or pyridazinyl; or a monocyclic heterocycle selected from quinolyl, quinoxalinyl, indolyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzofuranyl, benzothienyl, 2,3-dihydrobenzo[1,4] ... A bicyclic heterocycle of oxacyclohexenyl or benzo[1,3]dioxolyl, or a monocyclic saturated hydrocarbon group of 3-6 carbon atoms, or a bicyclic saturated hydrocarbon group of 6-12 carbon atoms; the carbon atoms on the monocyclic or bicyclic ring are independently and optionally substituted by 1-4 O, S, N or NH; any position of each monocyclic or bicyclic ring is substituted by 1, 2 or 3 substituents, and the substituents are halogen, haloalkyl, hydroxyl, alkyl or alkoxy;

[0018] Halogen is fluorine, chlorine, bromine or iodine;

[0019] A haloalkyl group is a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms, or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms, or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms connected to a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms; wherein one or more carbon atoms are substituted by one or more halogen atoms.

[0020] Further, R 1 、R 2 independently selected from hydrogen, alkyl;

[0021] Q 1 is aryl or Het; aryl or Het optionally replaced by one or more R 5 Replacement, R 5 is hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio or alkoxyalkyl;

[0022] Q 2 is an aryl group; an optional position of the aryl group is replaced by at least one R 6 Replacement, R 6 is hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio or alkoxyalkyl

[0023] Further, R 1 、R 2independently selected from hydrogen, methyl or ethyl;

[0024] Q 1 Het or an aromatic ring substituted with a substituent; Het is dihydrobenzo[b][1,4]dioxin, benzo[d][1,3]dioxane, phenyl, naphthyl, pyrrolyl, furyl, thienyl, pyridyl, pyrazinyl, or pyrimidinyl; the substituent is methyl, ethyl, 1-2 halogens, or trifluoromethyl;

[0025] Q 2 It is a phenyl group.

[0026] Further, R 1 、R 2 is hydrogen;

[0027] Q 1 is Het, an aromatic ring or an aromatic ring substituted with a substituent; Het is dihydrobenzo[b][1,4]dioxin, benzo[d][1,3]dioxane, or phenyl; the substituent is trifluoromethyl or halogen;

[0028] Q 2 It is a phenyl group.

[0029] Further, the compounds are: N-hydroxy-4-(((5-phenyl-1,3,4-oxadiazol-2-yl)thio)methyl)benzamide, N-hydroxy-4-(((5-(4-(trifluoromethyl)phenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)benzamide, 4-(((5-(4-chlorophenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide, N-hydroxy-4-(((5-(4-methoxyphenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)benzamide, 4-(((5-(4-fluorophenyl)-1,3,4-oxadiazol-2-yl)thio)methyl) -N-hydroxybenzamide, 4-(((5-(3,4-dichlorophenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide, 4-(((5-(3-chlorophenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide, 4-(((5-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide or 4-(((5-(Benzo[d][1,3]dioxan-5-yl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide.

[0030] Furthermore, a pharmaceutically acceptable salt is an acid addition salt formed by the compound and an acid or an acid salt of an inorganic base; the acid is hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid; the inorganic base is a salt with an alkaline metal cation, an alkaline earth metal cation or an ammonium cation.

[0031] The present invention also provides a pharmaceutical composition comprising the above compound or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0032] The present invention also provides an oxadiazole histone deacetylase / GSK-3β dual inhibitor, comprising the above compound or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0033] The present invention also provides the use of the above compound or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing and / or treating histone deacetylase / GSK-3β disorders.

[0034] Furthermore, the histone deacetylase / GSK-3β disorder is lung cancer, melanoma, liver cancer, kidney cancer, leukemia, prostate cancer, thyroid cancer, skin cancer, pancreatic cancer, ovarian cancer, testicular cancer, breast cancer, bladder cancer, gallbladder cancer, myelodysplastic syndrome, lymphoma, esophageal cancer, gastrointestinal cancer, astrocytoma, neuroblastoma, glioma, schwannoma, mesothelioma, non-insulin-dependent diabetes mellitus, or an autoimmune disease.

[0035] The present invention has the following beneficial effects:

[0036] 1. GSK (glycogen synthase kinase) and HDAC (histone deacetylase) dual-target inhibitors have been a hot topic in the research and development of anti-tumor drugs in recent years. Their advantages are mainly reflected in the following aspects: Synergistic anti-tumor effects. GSK inhibition (such as GSK-3β): Regulates signaling pathways such as Wnt / β-catenin and NF-κB, inhibits tumor cell proliferation and induces apoptosis. Enhances chemotherapy / radiotherapy sensitivity and overcomes drug resistance. HDAC inhibition: Activates tumor suppressor genes through epigenetic regulation (histone acetylation) to inhibit tumor growth. Induces cell cycle arrest and differentiation. Synergistic effect: The simultaneous action of two targets can block complementary pathways, reduce the risk of single-drug resistance, and enhance anti-tumor effects. The combination of the two can more comprehensively reverse the malignant phenotype of the tumor.

[0037] 2. Pharmacological test results show that the compounds of the present invention and their pharmaceutically acceptable salts have excellent inhibitory activity against HDAC6 and GSK3β; therefore, the compounds and their pharmaceutically acceptable salts can be used to treat clinical conditions related to the above targets. Diseases related to HDAC6 and GSK3β can include, but are not limited to: lung cancer, melanoma, liver cancer, kidney cancer, leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, pancreatic cancer, ovarian cancer, testicular cancer, breast cancer, bladder cancer, gallbladder cancer, myelodysplastic syndrome, lymphoma, esophageal cancer, follicular thyroid cancer, gastrointestinal cancer, tumors of the central or peripheral nervous system (such as astrocytoma, neuroblastoma, glioma or schwannoma), mesothelioma, type II or non-insulin-dependent diabetes mellitus, and autoimmune diseases. DETAILED DESCRIPTION

[0038] The principles and features of the present invention are described below. The examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. In the examples, where specific conditions are not specified, conventional conditions or manufacturer-recommended conditions were used. Reagents or instruments used where the manufacturer is not specified are conventional products that can be purchased commercially.

[0039] Example 1

[0040] N-hydroxy-4-(((5-phenyl-1,3,4-oxadiazol-2-yl)thio)methyl)benzamide, whose structural formula is

[0041] The preparation method comprises the following steps:

[0042] Step a: Preparation of 4-(chloromethyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide:

[0043] To a 50 mL single-necked flask were added 4-(chloromethyl)benzoic acid (100 mg, 0.267 mmol), HATU (93.6 mg, 0.292 mmol), DIPEA (0.184 mL, 1.06 mmol), and 25 mL of DMF. The mixture was stirred at room temperature for 30 min, followed by the addition of O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (28.66 mg, 0.265 mmol). Stirring was continued for 6 h until the reaction was complete. 100 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (100 mL × 3). The organic phase was collected, dried over anhydrous magnesium sulfate, and filtered to obtain a filtrate, which was purified by silica gel column chromatography (EA:MeOH=50:3) to obtain 70 mg of a white solid, with a yield of 57.3%.

[0044] Step b: Preparation of benzoylhydrazide:

[0045] Methyl benzoate (0.62 g, 4.59 mmol) and hydrazine hydrate (1.43 mL, 45.9 mmol) were reacted in ethanol (20 mL) at 90°C for 16-24 h. The reaction was complete by TLC. A solid precipitated in the solution, which was filtered and dried to obtain the title compound as a white solid in a 95% yield.

[0046] Step c: Preparation of 5-phenyl-1,3,4-oxadiazole-2-thiol:

[0047] Benzoylhydrazide (0.28 g, 2.05 mmol), CS2 (0.16 g, 2.05 mmol), and triethylamine (0.62 g, 6.15 mmol) were weighed and added to a 125 mL eggplant flask. Ethanol (25 mL) was then added dropwise. The mixture was heated to reflux and reacted for 8-12 hours. TLC confirmed the reaction was complete. Dilute hydrochloric acid was added dropwise until the solution became acidic, resulting in the precipitation of solids. The filter cake was filtered and dried to obtain the product as a dark green solid with a yield of 65%.

[0048] Step d: Preparation of 4-(((5-phenyl-1,3,4-oxadiazol-2-yl)thio)methyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide:

[0049] 5-Phenyl-1,3,4-oxadiazole-2-thiol (0.25 g, 1.40 mmol) and 4-(chloromethyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide (0.36 g, 1.33 mmol) were placed in a 50 mL dry two-necked flask. Under argon protection, NaH (0.04 g, 1.73 mmol) and DMF (7.5 mL) were added and reacted at room temperature for 6-8 h. The reaction was complete when detected by TLC. Post-treatment was performed as in step a. The title compound was isolated and purified by column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain a white solid in a yield of 63%.

[0050] Step e: Synthesis of N-hydroxy-4-(((5-phenyl-1,3,4-oxadiazol-2-yl)thio)methyl)benzamide:

[0051] 4-(((5-phenyl-1,3,4-oxadiazol-2-yl)thio)methyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide (0.18 g, 0.43 mmol) and p-toluenesulfonic acid (0.02 g, 0.13 mmol) were added to a 100 mL eggplant flask, dissolved in methanol, and reacted at room temperature for 16-24 h. The reaction was complete when detected by TLC. Dilute ammonia was added dropwise to adjust the pH to weak alkaline, and solid precipitated. The filter cake was filtered and dried to obtain the title compound in a yield of 69.6%.

[0052] Melting point 180.3-181.9℃. 1H NMR (400MHz, DMSO-d6) δ11.19(s,1H),9.03(s,1H),7.95(dd,J=8.0,1.5Hz,2H),7.71(d,J=8.3Hz,2H),7.64–7.54(m,5H),4.62(s,2H). 13 C NMR (101MHz, DMSO) δ165.35,163.89,163.20,140.01,132.20,132.13,129.51,129.10,127.18,126.45,122.99,35.44.HR-MS(ESI,m / z):Calcd for 328.0750.(C 16 H 14 N3O3S + [M+H] + ).Found328.0746

[0053] Example 2

[0054] N-hydroxy-4-(((5-(4-(trifluoromethyl)phenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)benzamide, whose structural formula is:

[0055] Using p-trifluoromethyl benzoate as raw material, the preparation method was similar to that in Example 1 to obtain a pale yellow solid with a yield of 67.3% and a melting point of 201.3-203.1°C. 1 H NMR (400MHz, DMSO-d6) δ11.19(s,1H),9.03(s,1H),8.17(d,J=8.2Hz,2H),7.9 7(d,J=8.3Hz,2H),7.72(d,J=8.2Hz,2H),7.57(d,J=8.2Hz,2H),4.65(s,2H). 13 C NMR(101MHz,DMSO)δ164.29,164.16,163.87,139.87,132.23,131.72,131.40,129.10,1 27.28,127.18,126.75,126.44,126.40,125.08,122.37,35.42.HR-MS(ESI,m / z):Calcd for

[0056] 396.0624.(C 17 H 13 F3N3O3S + [M+H] + ).Found 396.0621.

[0057] Example 3

[0058] 4-(((5-(4-chlorophenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide, whose structural formula is:

[0059] Using methyl p-chlorobenzoate as raw material, the preparation method refers to Example 1, and the product is a pale yellow solid with a yield of 60.3% and a melting point of 178.5-180.1°C. 1 H NMR (400MHz, DMSO-d6) δ11.19(s,1H),9.03(s,1H),7.96(d,J=8.6Hz,2H),7.69(dd,J=17.2,8.4Hz,4H),7.55(d,J=8.2Hz,2H),4.62(s,2H). 13 C NMR (101MHz, DMSO) δ164.58,163.87,163.47,139.93,136.79,132.20,129.64,129.08,128.24,127.16,121.87,35.42.HR-MS(ESI,m / z):Calcd for 384.0180.(C 16 H 12 NaCl 35 N3O3S + [M+Na] + ).Found384.0177.

[0060] Example 4

[0061] N-hydroxy-4-(((5-(4-methoxyphenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)benzamide, whose structural formula is:

[0062] Using methyl p-methoxybenzoate as raw material, the preparation method refers to Example 1, and the product is a white solid with a yield of 65.6% and a melting point of 153.2-154.9°C. 1 H NMR (400MHz, DMSO-d6) δ11.18(s,1H),9.03(s,1H),7.89(d,J=8.9Hz,2H),7.70(d,J =8.3Hz,2H),7.54(d,J=8.3Hz,2H),7.13(d,J=8.9Hz,2H),4.60(s,2H),3.85(s,3H). 13C NMR (101MHz, DMSO) δ165.29,164.01,163.88,162.32,162.11,158.62,142.64,140.05, 132.16,129.05,128.31,127.15,115.31,114.93,55.57,35.46.HR-MS(ESI,m / z):Calcd for 380.0675.(C 17 H 15 NaN3O4S + [M+Na] + ).Found 380.0670.

[0063] Example 5

[0064] 4-(((5-(4-fluorophenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide, whose structural formula is:

[0065] Using methyl p-fluorobenzoate as raw material, the preparation method refers to Example 1. White solid, yield 63.5%, melting point 189.7-191.4°C. 1 H NMR(400MHz,DMSO-d6)δ11.19(s,1H),9.04(s,1H),8.03(dd,J=8.9,5.3Hz,2H), 7.72(d,J=8.2Hz,2H),7.56(d,J=8.2Hz,2H),7.45(t,J=8.9Hz,2H),4.63(s,2H). 13 C NMR(101MHz,DMSO)δ165.39,164.60,163.88,163.20,139.96,132.19,129.23,129.14,129.07,127.16,116.85,116.63,35.43.HR-MS(ESI,m / z):Calcd for 368.0475.(C 16 H 12 NaFN3O3S + [M+Na] + ).Found 368.0472.

[0066] Example 6

[0067] 4-(((5-(3,4-dichlorophenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide, whose structural formula is:

[0068] Using methyl 3,4-dichlorobenzoate as the starting material, the preparation method was similar to that of Example I-1. The product was a pale yellow solid with a yield of 64.8% and a melting point of 183.6-185.1°C. 1 H NMR (400MHz, DMSO-d6) δ11.19(s,1H),9.04(s,1H),8.15(d,J=1.9Hz,1H),7.93(dd,J=8.4,2 .0Hz,1H),7.87(d,J=8.4Hz,1H),7.71(d,J=8.2Hz,2H),7.56(d,J=8.2Hz,2H),4.64(s,2H). 13 C NMR(101MHz,DMSO)δ163.98,163.65,139.93,134.78,132.38,132.21,131.87,129.12,128.12,127.17,126.54,123.48,35.38.HR-MS(ESI,m / z):Calcd for 417.9790.(C 16 H 11 NaCl 35 2N3O3S + [M+Na] + ).Found 417.9787.

[0069] Example 7

[0070] 4-(((5-(3-chlorophenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide, whose structural formula is:

[0071] Using methyl 3-chlorobenzoate as raw material, the preparation method refers to Example 1. Pale yellow solid, yield 66.9%, melting point 182.1-183.8°C. 1 H NMR(400MHz,DMSO-d6)δ11.20(s,1H),9.05(s,1H),7.97–7.91(m,2H),7.71 (d,J=8.2Hz,3H),7.63(t,J=7.9Hz,1H),7.56(d,J=8.2Hz,2H),4.63(s,2H). 13 C NMR(101MHz,DMSO)δ164.23,163.78,139.97,134.12,132.21,131.92,131.55,129.12,127.18,125.99,125.15,124.93,35.39.HR-MS(ESI,m / z):Calcd for 384.0180.(C 16 H12 NaCl 35 N3O3S + [M+Na] + ).Found 384.0175.

[0072] Example 8

[0073] 4-(((5-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide, whose structural formula is:

[0074]

[0075] Using 2,3-dihydrobenzo[b][1,4]dioxin-6-carboxylic acid methyl ester as raw material, the preparation method refers to Example 1, and the product is a white solid with a yield of 58.6% and a melting point of 176.2-177.7°C. 1 H NMR (400MHz, DMSO-d6) δ11.20(s,1H),9.05(s,1H),7.70(d,J=8.2Hz,2H),7.54(d,J=8. 2Hz,2H),7.44–7.39(m,2H),7.05(d,J=8.4Hz,1H),4.59(s,2H),4.32(d,J=4.0Hz,4H). 13 C NMR (101MHz, DMSO) δ165.00,163.86,162.49,146.76,143.86,140.03,132.15,129.03, 127.13,119.99,118.20,115.91,115.09,64.46,64.09,35.41.HR-MS(ESI,m / z):Calcd for 386.0805.(C 18 H 16 N3O5S + [M+H] + ).Found386.0800.

[0076] Example 9

[0077] 4-(((5-(Benzo[d][1,3]dioxane-5-yl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide, whose structural formula is:

[0078]

[0079] Using benzo[d][1,3]dioxolane-5-carboxylic acid methyl ester as raw material, the preparation method refers to Example 1. Pale yellow solid, yield 62.9%, melting point 186.5-188.2°C. 1 H NMR (400MHz, DMSO-d6) δ11.18(s,1H),9.02(s,1H),7.70(d,J=8.2Hz,2H),7.56–7. 48(m,3H),7.44(d,J=1.6Hz,1H),7.11(d,J=8.2Hz,1H),6.16(s,2H),4.60(s,2H). 13 C NMR (101MHz, DMSO) δ165.15,163.87,162.51,150.51,148.16,140.04,132.17,129. 06,127.15,121.77,116.63,109.20,106.18,102.18,35.41.HR-MS(ESI,m / z):Calcd for 372.0648.(C 17 H 14 N3O5S + [M+H] + ).Found 372.0642.

[0080] Test example

[0081] (1) Determination of the inhibitory activity of target compounds on HDAC6

[0082] The inhibitory activity of the compounds obtained in Examples 1-9 on HDAC was determined by fluorescence resonance energy transfer (FRET) and compared with the positive control drug to screen out compounds with better activity. HDAC was obtained by purification or direct purchase of a kit.

[0083] Specific method: Add enzyme to the reaction wells and reaction buffer to the control wells. Add samples dissolved in DMSO to the reaction wells and incubate using a non-contact nanoliter acoustic pipetting system. Add the corresponding fluorescent substrate to each reaction well, rotate and shake, seal and incubate at 30°C for 1-2 hours. Add a color developer containing TMP26 to terminate the reaction and generate fluorescence. Use the EnVision multi-label microplate reader (Perkin Elmer) to detect fluorescence intensity (excitation light: 490nM, emission light: 520nM). Read the endpoint value after the color development reaches stability. Use GraphPad Prism 4 software to calculate the percentage (relative to the DMSO control group) and half-maximal inhibition rate.

[0084] (2) Determination of the inhibitory activity of the target compound on GSK-3β enzyme

[0085] Experimental steps: Purified GSK-3β enzyme (0.5 μg) was mixed with reaction buffer (50 mM Tris-HCl, pH 7.3, 10 mM magnesium acetate (MgAc2), 0.01% β-mercaptoethanol). 32 P-γ-ATP (100 μM, 0.5 μCi / reaction) and peptide substrate pIRS-1 (100 μM, sequence: RREGGMSRPAS(p)VDG). The test compound was added to the reaction system at different concentrations (1, 10, 100 μM). Reaction conditions: Incubate at 30°C for 15 min. After the reaction is terminated, the mixture is spotted on a Whatman P81 phosphocellulose filter. The filter membrane is washed with 10 mM phosphoric acid solution to remove free 32P-ATP. The 32P-labeled substrate bound to the filter membrane is detected by radioactivity counting. Activity calculation: Taking the GSK-3β activity of the group without inhibitor as 100%, the percentage of inhibition of the enzyme activity by each concentration of compound is calculated.

[0086] The above results are shown in Table 1.

[0087] Table 1 Inhibitory activity of compounds on enzymes (IC 50 , nM)

[0088]

[0089]

[0090] As shown in Table 1, the compounds of the present invention exhibit dual-target inhibitory activity. Examples 4 and 9 have comparable HDAC activity to the positive drug SAHA, while exhibiting significant inhibitory activity against GSK-3β, superior to SAHA.

[0091] (3) Determination of the in vitro antitumor activity of the target compound

[0092] In this experiment, the CTG method was used to determine the inhibitory effect of the compounds on the gastric cancer cell line AGS.

[0093] Experimental method: Collect cells in the exponential growth phase for viable cell counting. Adjust the cell suspension concentration with the corresponding culture medium of each cell. Add 90 μL of cell suspension to each well of a 96-well cell culture plate. Dissolve each test compound in DMSO to a 10mM or 5mM stock solution. Then dilute to a 10-fold solution with culture medium, and repeat each well. Add 10 μL of the corresponding 10-fold solution to each well of each cell line, with a final drug concentration of 10 μM or 25 μM, and a final DMSO concentration of 0.1%-0.5% (see compound preparation method and sample addition design: experimental well plate sample addition design). Culture in a 37°C, 5% CO2 incubator for 72 hours. After 72 hours of drug treatment, add 50 μL (1 / 2 culture volume) of CTG solution that has been melted and equilibrated to room temperature to each well, mix with a microplate shaker for 2 minutes, and place at room temperature for 10 minutes. Measure the fluorescence signal value using an Envision2104 plate reader. The cell inhibition rate is calculated using the formula: (1-V sample / V vehicle control × 100%). sample is the mean value of the drug-treated group, V vehicle control The results are shown in Table 2.

[0094] Table 2 Antiproliferative activity of compounds against AGS tumor cell lines (IC 50 , μM)

[0095]

[0096]

[0097] As shown in Table 2, some examples of the present invention exhibited excellent anti-proliferative activity, among which Examples 4 and 8 were significantly more active than the positive control drugs SAHA and ACY1215.

[0098] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, characterized in that: The general structural formula of the compound is: Among them, R 1 、R 2 are independently selected from hydrogen, alkyl, alkenyl, or alkynyl; Q 1 is aryl or Het; aryl or Het is optionally replaced by at least one R 3 Replacement, R 3 is hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl or Het; Q 2 is aryl or Het; aryl or Het is optionally replaced by at least one R 4 Replacement, R 4 is hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylamino, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl or Het; The alkyl group is a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms connected to a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms; The alkoxy group is a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms connected to a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms; wherein each carbon atom is optionally substituted by oxygen; The alkylamino group is a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms connected to a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms; wherein each carbon atom is optionally substituted by an NH group; The alkoxyalkyl group is the alkoxy group connected to the alkyl group; The alkenyl or alkynyl group is a straight or branched unsaturated hydrocarbon group containing double bonds or triple bonds having 1 to 6 carbon atoms; The aryl group is a carbon ring of phenyl, naphthyl, acenaphthenyl or tetrahydronaphthyl, optionally substituted by 1, 2 or 3 substituents, wherein the substituents are hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxyl, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl or Het; The aralkyl or diarylalkyl group is the aryl group connected to the alkyl group; Het is a monocyclic heterocyclic ring selected from pyrrolyl, pyrazolyl, imidazolyl, furyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl or pyridazinyl; or selected from quinolyl, quinoxalinyl, indolyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzofuranyl, benzothienyl, 2,3-dihydrobenzo[1,4] A bicyclic heterocycle of a dioxinyl or benzo[1,3]dioxolyl group, or a monocyclic saturated hydrocarbon group of 3-6 carbon atoms, or a bicyclic saturated hydrocarbon group of 6-12 carbon atoms; the carbon atoms on the monocyclic or bicyclic ring are independently and optionally substituted by 1-4 O, S, N or NH; any position of each monocyclic or bicyclic ring is substituted by 1, 2 or 3 substituents, and the substituents are halogen, haloalkyl, hydroxyl, alkyl or alkoxy; The halogen is fluorine, chlorine, bromine or iodine; The haloalkyl group is a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms, or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms, or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms connected to a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms; wherein one or more carbon atoms are substituted by one or more halogen atoms.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 、R 2 independently selected from hydrogen, alkyl; Q 1 is aryl or Het; aryl or Het optionally replaced by one or more R 5 Replacement, R 5 is hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio or alkoxyalkyl; Q 2 is an aryl group; an optional position of the aryl group is replaced by at least one R 6 Replacement, R 6 is hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio or alkoxyalkyl.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 、R 2 independently selected from hydrogen, methyl or ethyl; Q 1 Het or an aromatic ring substituted with a substituent; Het is dihydrobenzo[b][1,4]dioxin, benzo[d][1,3]dioxane, phenyl, naphthyl, pyrrolyl, furyl, thienyl, pyridyl, pyrazinyl, or pyrimidinyl; the substituent is methyl, ethyl, 1-2 halogens, or trifluoromethyl; Q 2 It is a phenyl group.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 、R 2 is hydrogen; Q 1 is Het, an aromatic ring or an aromatic ring substituted with a substituent; Het is dihydrobenzo[b][1,4]dioxin, benzo[d][1,3]dioxane, or phenyl; the substituent is trifluoromethyl or halogen; Q 2 It is a phenyl group.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein The compounds are: N-hydroxy-4-(((5-phenyl-1,3,4-oxadiazol-2-yl)thio)methyl)benzamide, N-hydroxy-4-(((5-(4-(trifluoromethyl)phenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)benzamide, 4-(((5-(4-chlorophenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide, N-hydroxy-4-(((5-(4-methoxyphenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)benzamide, 4-(((5-(4-fluorophenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)- N-hydroxybenzamide, 4-(((5-(3,4-dichlorophenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide, 4-(((5-(3-chlorophenyl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide, 4-(((5-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide or 4-(((5-(Benzo[d][1,3]dioxan-5-yl)-1,3,4-oxadiazol-2-yl)thio)methyl)-N-hydroxybenzamide.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein The pharmaceutically acceptable salt is an acid addition salt formed by the compound and an acid or an acid salt of an inorganic base; the acid is hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid; the inorganic base is a salt with an alkaline metal cation, an alkaline earth metal cation or an ammonium cation.

7. A pharmaceutical composition, characterized in that The invention comprises the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

8. An oxadiazole histone deacetylase / GSK-3β dual inhibitor, characterized in that: The invention comprises the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

9. Use of the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating histone deacetylase / GSK-3β disorders.

10. The use according to claim 9, characterized in that The histone deacetylase / GSK-3β disorder is lung cancer, melanoma, liver cancer, kidney cancer, leukemia, prostate cancer, thyroid cancer, skin cancer, pancreatic cancer, ovarian cancer, testicular cancer, breast cancer, bladder cancer, gallbladder cancer, myelodysplastic syndrome, lymphoma, esophageal cancer, gastrointestinal cancer, astrocytoma, neuroblastoma, glioma, schwannoma, mesothelioma, non-insulin-dependent diabetes mellitus, or an autoimmune disease.

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