Compounds targeting the inhibition of tcptp and uses thereof

Abstract

The application discloses a compound for targeting and inhibiting TCPTP and application thereof. The compound has a molecular formula of (R)-N-(1-(3-(benzyl)-1-methyl-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)piperidin-3-yl)acetamide. The compound provided by the application has a brand-new structure, and has further development potential for developing TCPTP inhibitors and improving selectivity of the TCPTP inhibitors, and has good clinical application prospects for resisting tumors and reducing blood sugar. In addition, the novel compound provided by the application has a simple synthesis method, high yield and low content of by-products, and reduces production cost.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medicine, in particular to a compound for targeting and inhibiting TCPTP and application thereof. BACKGROUND

[0002] T cell protein tyrosine phosphatase (TCPTP) is an intracellular protein tyrosine phosphatase (PTPs) encoded by protein tyrosine phosphatase non-receptor type 2 (PTPN2) gene, which is expressed in various tissues and cells at all stages.

[0003] TCPTP plays a role in autoimmune diseases, cancer and cancer immunosurveillance, and is an attractive target for immune-mediated diseases. Various evidences show that activating TC-PTP is a candidate target for treating autoimmune diseases. Studies have also found that TCPTP is highly expressed in various tumor cells, and knocking down or knocking out TCPTP can inhibit the occurrence and development of tumors. In order to support TC-PTP as an immunotherapy target, two orally bioavailable TC-PTP inhibitors ABBV-CLS-579 and ABBV-CLS-484 (patent number WO / 2019 / 246513) developed by Calico and AbbVie are currently undergoing phase I clinical trials for locally advanced or metastatic tumors.

[0004] However, due to the high conservation between TCPTP and PTP1B (tyrosine protein phosphatase), it has always been challenging to identify TCPTP selective molecules with good pharmacological characteristics. Structural studies have shown that the two enzymes are allosterically regulated by their C-termini through different mechanisms, which suggests that they can be selectively targeted by approaching regions outside the catalytic domain. The compound provided by the present application has a completely new structure, which is different from the compound type reported in the literature, and has further development potential for developing TCPTP inhibitors and improving their selectivity. SUMMARY

[0005] In view of the deficiencies of the prior art, the present application provides a compound for targeting and inhibiting TCPTP and application thereof. The compound provided by the present application has a strong inhibitory effect on TCPTP and can be used for preparing an antitumor or hypoglycemic drug.

[0006] To solve the above technical problems, the technical scheme of the present application is as follows:

[0007] In the first aspect of the present application, a compound for targeting and inhibiting TCPTP is provided, and the structure of the compound is as follows:

[0008]

[0009] In the formula, R is selected from one or more of F, Cl, Br, cyano, alkyl and alkoxy.

[0010] As a preferred solution to the above-mentioned solution, the compound is selected from one of the following structural formulas:

[0011] The preferred compound has a strong inhibitory effect on TCPTP.

[0012] In the second aspect of the present application, the use of the compound in the preparation of a TCPTP inhibitor is further provided.

[0013] In the third aspect of the present application, the use of the compound in the preparation of an antitumor drug is further provided.

[0014] In the third aspect of the present application, the use of the compound in the preparation of a hypoglycemic drug is further provided.

[0015] Compared with the prior art, the present application has the following beneficial effects: the novel compound provided by the present application has a simple synthesis method, high yield and low by-product; the obtained compound has a strong inhibitory effect on TCPTP. It has good clinical application prospect for anti-tumor and hypoglycemic. DETAILED DESCRIPTION

[0016] The technical solutions of the present application will be further described in detail below in combination with specific embodiments, but the present application is not limited to the following technical solutions.

[0017] Synthesis of compound V and its application

[0018] The synthesis route of compound V is as follows:

[0019]

[0020] (1) Compound I and compound II are put into a reaction kettle according to a molar ratio of 1:1.0-1.1, and 1.05-1.2 molar ratio of diisopropyl ethylamine is added. Unless otherwise specified, the amount of compound I is 1. The temperature is raised to 65-75℃, and the reaction is stirred for 6-10 hours. After cooling to 0-5℃, filtration is performed. The filter cake is washed with tetrahydrofuran, and the wet filter cake is added with purified water. After being stirred at room temperature for 2-3 hours, filtration is performed, and the filter cake is washed with purified water and dried to obtain compound III.

[0021] (2) Compound III and (R)-3-aminopiperidine dihydrochloride are added into the kettle according to a molar ratio of 1:1.0-1.2, and no special indication is given. The amount of substance of the compound III is 1, and 2.1-2.4 moles of sodium carbonate are added, and ethanol is added as the solvent. The temperature is raised to 75-85°C, and the reaction is stirred for 5-6 hours. After the reaction is completed, the ethanol is concentrated, and filtration is performed to obtain compound IV.

[0022] (3) Compound IV and acetyl chloride are added into the reaction kettle according to a molar ratio of 1:1.1-1.3, and no special indication is given. The amount of compound IV is 1. The reaction is stirred at room temperature for 5-6 hours. After the reaction is completed, saturated aqueous sodium bicarbonate solution is added, and the organic phase is washed with water and then separated and concentrated. The organic phase is slurried in ethanol overnight, the filter cake is filtered, and dried to obtain compound V, with a yield of 56%.

[0023] The hydrogen spectrum nuclear magnetic data and mass spectrum detection data of compound V are as follows:

[0024] 1 H-NMR (DMSO): 10.73 (s, 1H), 7.70 (d, 1H), 7.51 (dd, 1H), 7.30 (dd, 1H), 7.18 (d, 1H), 6.75 (d, 1H), 3.76 (s, 2H), 3.14-3.12 (br, 2H), 3.06 (s, 3H), 3.01-2.98 (br, 2H), 2.79-2.56 (br, 2H), 1.70-1.54 (br, 2H), 1.48-1.37 (br, 1H), 1.29 (s, 3H); MS + = 400.4.

[0025] Non-receptor tyrosine-specific phosphatase (TCPTP) inhibition activity determination

[0026] The test compounds I-V are respectively prepared into test solution of different concentrations with DMSO, and 2 μL of the test solution is respectively added into the standard activity determination system (50 mM Tris-HCl, pH 6.5, 2 mM pNPP, 2% DMSO, 30 nM hGST-PTP1B). The negative control is DMSO, and the positive control is sodium orthovanadate. The reaction temperature is 30°C, the dynamic determination wavelength is 405 nm, the time is 3 min, and the inhibition rate of the compound on the TCPTP enzyme activity is calculated according to the following formula. Inhibition rate = (experimental group A value-negative control group A value) / (control group A value-negative control group A) x 100%, and the results are shown in Table 1.

[0027] Table 1 TCPTP inhibition activity determination

[0028]

[0029] If the inhibition rate is greater than 50% at a compound concentration of 20 μg / mL, then continue to test at a concentration level of 5 μg / mL.

[0030] The test results show that the compound V has a significant inhibitory effect on non-receptor tyrosine-specific phosphatase, and has a good clinical application prospect for anti-tumor and blood glucose reduction.

[0031] Synthesis of compound 5 of example 2 and its application

[0032] The synthesis route of compound 5 is as follows:

[0033]

[0034] (1) Compound 1 and compound 2 are put into the reaction kettle according to the molar ratio of 1:1.0-1.2, and 1.0-1.2 molar ratio of diisopropyl ethylamine is added. Unless otherwise specified, compound 1 is unit 1. The temperature is raised to 65-75℃, and the reaction is stirred for 6-10 hours. After cooling to 0-5℃, filter. The filter cake is washed with tetrahydrofuran, and the wet filter cake is added with purified water. The pulp is beaten at room temperature for 2-3 hours, filtered, washed with purified water and dried to obtain compound 3.

[0035] (2) Compound 3 and (R)-3-aminopiperidine dihydrochloride are added to the kettle according to the molar ratio of 1:1.1-1.3. Unless otherwise specified, compound 3 is unit 1. Molar ratio of 2.1-2.3 sodium carbonate is added, and ethanol is added. The temperature is raised to 75-85℃, and the reaction is stirred for 5-6 hours. After the reaction is completed, the ethanol is concentrated and filtered.

[0036] (3) Compound 4 and acetyl chloride are put into the reaction kettle according to the molar ratio of 1:1.1-1.3, and appropriate volume of dichloromethane is added. Unless otherwise specified, compound 4 is unit 1. The reaction is stirred at room temperature for 5-6 hours. After the reaction is completed, saturated aqueous sodium bicarbonate solution is added, and the organic phase is washed with water and concentrated. The organic phase is added with ethanol to pulp overnight, filtered, and dried to obtain compound 5 with a yield of 55%.

[0037] The hydrogen spectrum nuclear magnetic data and mass spectrum detection data of compound 5 are as follows:

[0038] 1H-NMR (DMSO): 8.14 (s, 1H), 7.64 (d, 1H), 7.51 (dd, 1H), 7.30 (dd, 1H), 7.18 (d, 1H), 6.75 (d, 1H), 3.76 (s, 2H), 3.14-3.12 (br, 2H), 3.06 (s, 3H), 3.01-2.98 (br, 2H), 2.79-2.56 (br, 2H), 2.29 (s, 3H), 1.99 (s, 3H), 1.70-1.54 (br, 2H), 1.48-1.37 (br, 1H); MS + = 389.4.

[0039] The inhibition rates of compounds 1-5 on TCPTP inhibitory activity are shown in Table 2.

[0040] Table 2 TCPTP inhibitory activity assay

[0041]

[0042] If the inhibition rate is about 50% at a compound concentration of 20 μg / mL, then the test at a concentration level of 5 μg / mL is continued.

[0043] The test results show that compound 5 has a significant inhibitory effect on non-receptor tyrosine-specific phosphatase, and has a good clinical application prospect for anti-tumor and blood glucose reduction.

[0044] Synthesis of compound E and its application

[0045] The synthesis route of compound E is as follows:

[0046]

[0047] (1) Compound A and compound B are put into a reaction kettle at a molar ratio of 1:1.1-1.2, and 1.20-1.40 molar ratio of diisopropyl ethylamine is added. Unless otherwise specified, the amount of compound A is 1 unit. The temperature is raised to 65-75°C, and the reaction is stirred for 6-10 hours. After cooling to 0-5°C, the filter cake is washed with tetrahydrofuran, and the wet filter cake is added with purified water, and the pulp is beaten at room temperature for 2-3 hours, filtered, washed with purified water and dried to obtain compound C.

[0048] (2) Compound C and (R)-3-aminopiperidine dihydrochloride are added to the kettle at a molar ratio of 1:1.1-1.3, and 2.8-3.2 molar ratio of sodium carbonate is added. The temperature is raised to 75-85°C, and the reaction is stirred for 5-6 hours. After the reaction is completed, the ethanol is concentrated and filtered.

[0049] (3) Compound D and acetyl chloride are put into a reaction kettle in a molar ratio of 1:1.1-1.2, and a proper volume of dichloromethane is added. Unless otherwise specified, the amount of substance of Compound D is 1 unit. Stirring is carried out at room temperature for 5-6 hours. After the reaction is completed, saturated aqueous sodium bicarbonate solution is added, and the organic phase is washed with water and then separated and concentrated. The organic phase is slurried in ethanol overnight, the filter cake is filtered, and the filter cake is dried to obtain Compound E, with a yield of 58%.

[0050] The hydrogen spectrum nuclear magnetic data and mass spectrum detection data of Compound E are as follows:

[0051] 1 H-NMR (DMSO): 8.14 (s, 1H), 7.64 (d, 1H), 7.51 (dd, 1H), 7.30 (dd, 1H), 7.18 (d, 1H), 6.75 (d, 1H), 3.76 (s, 2H), 3.14-3.12 (br, 2H), 3.06 (s, 3H), 3.01-2.98 (br, 2H), 2.79-2.56 (br, 2H), 2.29 (s, 3H), 1.99 (s, 3H), 1.70-1.54 (br, 2H), 1.48-1.37 (br, 1H); MS + = 396.4.

[0052] Non-receptor tyrosine-specific phosphatase inhibition activity assay

[0053] The test compounds A-E are respectively prepared into test solution of different concentrations with DMSO, 2 μL of the test solution is respectively added into a standard activity assay system (50 mM Tris-HCl, PH 6.5, 2 mM pNPP, 2% DMSO, 30 nM hGST-PTP1B), the negative control is DMSO, and the positive control is sodium orthovanadate. The reaction temperature is 30°C, the dynamic determination wavelength is 405 nm, the time is 3 min, and the inhibition rate of the TCPTP enzyme activity of the compound is calculated according to the following formula: inhibition rate = (experimental group A value-negative control group A value) / (control group A value-negative control group A) * 100%, and the results are shown in Table 3.

[0054] Table 3 TCPTP inhibition activity assay

[0055]

[0056] If the inhibition rate is about 50% when the concentration of the compound is 20 μg / mL, the 5 μg / mL concentration level test is continued.

[0057] The test results show that Compound E has a significant inhibitory effect on non-receptor tyrosine-specific phosphatase, and has good clinical application prospects for reducing blood sugar and resisting tumors.

[0058] The hypothalamic phosphatase TCPTP activity is closely related to fat metabolism: when fasting, TCPTP is activated, thereby inhibiting the insulin signal of the brain AgRP / NPY neurons, preventing the "browning" of white fat, and reducing energy consumption. In turn, food intake reduces the TCPTP enzyme activity in the hypothalamus, thereby up-regulating the insulin signal, promoting fat browning, and accelerating energy consumption.

[0059] Insulin can act in peripheral tissues and the brain to regulate glucose metabolism. The insulin receptor signaling pathway can inhibit AgRP neurons in the hypothalamus to promote insulin inhibition of hepatic glycogen synthesis in the periphery, and activation of AgRP neurons can impair the glucose uptake capacity of brown adipose tissue. Previous studies have found that tyrosine phosphatase TCPTP can inhibit the insulin receptor signal of AgRP neurons, and TCPTP in the hypothalamus is induced to express under fasting conditions, and TCPTP is degraded after eating. TCPTP can control the insulin receptor signal of AgRP neurons, coordinating the hepatic glycogen synthesis and glucose uptake response of brown / beige adipocytes to feeding and fasting.

[0060] The above are only preferred embodiments of the present application, and the protection scope of the present application is not limited to the above-mentioned embodiments. Any modification, equivalent replacement or improvement made by those skilled in the art without departing from the principles of the present application shall be considered as falling within the protection scope of the present application.

Claims

1. A compound which targets the inhibition of TCPTP, characterized in that, The compound is selected from one of the following structural formulas: or .

2. Use of a compound according to claim 1 for the preparation of a TCPTP inhibitor.

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