Compound for targeted inhibition of TCPTP and application thereof

Abstract

The invention discloses a compound for targeted inhibition of TCPTP and application of the compound. The molecular formula of the compound is (R)-N-(1-(3-(benzyl)-1-methyl-2, 6-dioxo-1, 2, 3, 6-tetrahydropyrimidine-4-yl) piperidine-3-yl) acetamide, and the structural formula of the compound is shown in the specification. The compound provided by the invention has a brand-new structure, has a further development potential for developing a TCPTP inhibitor and improving the selectivity of the TCPTP inhibitor, and has a good antitumor and hypoglycemic clinical application prospect. In addition, the novel compound provided by the invention is simple in synthesis method, high in yield and low in byproduct content, and the production cost is reduced.

Description

Technical Field

[0001] The present invention relates to the field of medicines, and in particular to a compound for targeted inhibition of TCPTP and application thereof. Background Art

[0002] T cell protein tyrosine phosphatase (TCPTP) is an intracellular protein tyrosine phosphatase (PTPs) encoded by the protein tyrosine phosphatase non-receptor type 2 (PTPN2) gene and is expressed in various tissues and cells throughout the body 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 indicate that activation of TC-PTP is a candidate target for the treatment of autoimmune diseases. Studies have also found that TCPTP is highly expressed in a variety of tumor cells, and knocking down or knocking out TCPTP can inhibit the occurrence and development of tumors. To support TC-PTP as an immunotherapy target, two orally bioavailable TC-PTP inhibitors ABBV-CLS-579 and ABBVCLS-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 conservatism between TCPTP and PTP1B (tyrosine protein phosphatase), it is always challenging to identify TCPTP selective molecules with good pharmacological characteristics. Structural studies have shown that these two enzymes are allosterically regulated by different mechanisms through their C-terminals, which indicates that they may be selectively targeted by approaching the region outside the catalytic domain. Compounds provided by the invention have brand-new structures, are different from the compound types reported in the literature, and have the potential for further development to the development of TCPTP inhibitors and to improve their selectivity. Summary of the invention

[0005] In view of the deficiencies of the prior art, the present invention provides a compound for targeted inhibition of TCPTP and its application. The compound provided by the present invention has a strong inhibitory effect on TCPTP and can be used to prepare anti-tumor or hypoglycemic drugs.

[0006] In order to solve the above technical problems, the technical solution of the present invention is as follows:

[0007] In a first aspect, the present invention provides a compound for targeted inhibition of TCPTP, the compound having the following structural formula:

[0008]

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

[0010] As a preferred embodiment of the above scheme, the compound is selected from one of the following structural formulas;

[0011] The preferred compounds have a strong inhibitory effect on TCPTP.

[0012] The second aspect of the present invention also provides the use of the compound in the preparation of TCPTP inhibitors.

[0013] The third aspect of the present invention also provides the use of the compound in the preparation of anti-tumor drugs.

[0014] The third aspect of the present invention also provides the use of the compound in the preparation of hypoglycemic drugs.

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

[0016] The technical solution of the present invention is further described in detail below in conjunction with specific embodiments, but the present invention is not limited to the following technical solution.

[0017] Example 1 Synthesis and Application of Compound V

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

[0019]

[0020] (1) Compound I and Compound II are put into a reaction kettle at a molar ratio of 1:1.0-1.1, and diisopropylethylamine is added at a molar ratio of 1.05-1.2. Unless otherwise specified, the molar unit of Compound I is 1. The temperature is raised to 65-75°C for reaction, and after stirring for 6-10 hours, the mixture is cooled to 0-5°C and filtered. The filter cake is rinsed with tetrahydrofuran, purified water is added to the wet filter cake, and the mixture is slurried at room temperature for 2-3 hours, filtered, rinsed with purified water, and dried to obtain Compound III.

[0021] (2) Compound III and (R)-3-aminopiperidine dihydrochloride are added to a kettle at a molar ratio of 1:1.0-1.2. Unless otherwise specified, the amount of compound III in this step is 1. Sodium carbonate is added at a molar ratio of 2.1-2.4, and ethanol is added as 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 filtered to obtain compound IV.

[0022] (3) Compound IV and acetyl chloride were added to a reactor in a molar ratio of 1:1.1-1.3, and an appropriate volume of dichloromethane was added. Unless otherwise specified, the unit volume of compound IV was 1. The reaction was stirred at room temperature for 5-6 hours. After the reaction, a saturated aqueous solution of sodium bicarbonate was added, and the organic phase was washed with water and then separated and concentrated. Ethanol was added to the organic phase to pulp overnight, and the filter cake was filtered and dried to obtain compound V, with a yield of 56%.

[0023] The H NMR data and mass spectrometry 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),3.14-3.1 MS + =400.4.

[0025] Non-receptor tyrosine-specific phosphatase (TCPTP) inhibitory activity assay

[0026] Compound IV to be tested was prepared into test solutions of different concentrations with DMSO, and 2 μL of the test solution was added to the standard activity test system (50 mM Tris-HCl, pH 6.5, 2 mM pNPP, 2% DMSO, 30 nM hGST-PTP1B), negative control: DMSO; positive control: sodium orthovanadate. The reaction temperature was 30 ° C, the dynamic measurement wavelength was the light absorption at 405 nm, the time was 3 min, and the inhibition rate of the compound on TCPTP enzyme activity was 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%, the results are shown in Table 1.

[0027] Table 1 TCPTP inhibitory activity assay

[0028]

[0029] When the compound concentration was 20 μg / mL, if the inhibition rate was greater than 50%, the test was continued at the 5 μg / mL concentration level.

[0030] The experimental results show that compound V exhibits a significant inhibitory effect on non-receptor tyrosine-specific phosphatases and has good clinical application prospects in anti-tumor and hypoglycemic effects.

[0031] Example 2 Synthesis and Application of Compound 5

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

[0033]

[0034] (1) Compound 1 and Compound 2 are put into a reaction kettle at a molar ratio of 1:1.0-1.2, and diisopropylethylamine is added at a molar ratio of 1.0-1.2. Unless otherwise specified, Compound 1 is 1. The temperature is raised to 65-75°C for reaction, and after stirring for 6-10 hours, the mixture is cooled to 0-5°C and filtered. The filter cake is rinsed with tetrahydrofuran, purified water is added to the wet filter cake, and the mixture is slurried at room temperature for 2-3 hours, filtered, rinsed with purified water, and dried to obtain Compound 3.

[0035] (2) Compound 3 and (R)-3-aminopiperidine dihydrochloride are added to the kettle at a molar ratio of 1:1.1-1.3. Unless otherwise specified, the compound 3 is 1 in this step. Sodium carbonate is added at a molar ratio of 2.1-2.3, and ethanol 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.

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

[0037] The H NMR data and mass spectrometry 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,2 MS + =389.4.

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

[0040] Table 2 TCPTP inhibitory activity assay

[0041]

[0042] When the compound concentration is 20 μg / mL, if the inhibition rate is around 50%, the test will continue at the 5 μg / mL concentration level.

[0043] The experimental results show that compound 5 exhibits a significant inhibitory effect on non-receptor tyrosine-specific phosphatases and has good clinical application prospects in anti-tumor and hypoglycemic activities.

[0044] Example 3 Synthesis and Application of Compound E

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

[0046]

[0047] (1) Compound A and compound B are put into a reactor at a molar ratio of 1:1.1-1.2, and diisopropylethylamine is added at a molar ratio of 1.20-1.40. Unless otherwise specified, the amount of compound A is 1. The temperature is raised to 65-75°C for reaction, and after stirring for 6-10 hours, the mixture is cooled to 0-5°C and filtered. The filter cake is rinsed with tetrahydrofuran, purified water is added to the wet filter cake, and the mixture is slurried at room temperature for 2-3 hours, filtered, rinsed with purified water, and dried to obtain compound C.

[0048] (2) Compound C and (R)-3-aminopiperidine dihydrochloride are added to a kettle at a molar ratio of 1:1.1-1.3, and sodium carbonate and ethanol are added at a molar ratio of 2.8-3.2. 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 were added to a reaction kettle at a molar ratio of 1:1.1-1.2, and an appropriate volume of dichloromethane was added. Unless otherwise specified, the amount of compound D was 1. The reaction was stirred at room temperature for 5-6 hours. After the reaction was completed, a saturated aqueous solution of sodium bicarbonate was added, and the organic phase was washed with water and then separated and concentrated. Ethanol was added to the organic phase to pulp overnight, and the filter cake was filtered and dried to obtain compound E, with a yield of 58%.

[0050] The hydrogen nuclear magnetic resonance data and mass spectrometry 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,2 MS + =396.4.

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

[0053] Compound A-Compound E to be tested were prepared into test solutions of different concentrations with DMSO, and 2 μL of the test solutions were added to the standard activity test system (50 mM Tris-HCl, pH 6. 5.2 mM pNPP, 2% DMSO, 30 nM hGST-PTP1B), negative control: DMSO, positive control: sodium orthovanadate, reaction temperature was 30°C, dynamic measurement wavelength was light absorption at 405 nm, time was 3 min, and the TCPTP enzyme activity inhibition rate of the compound was 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%, the results are shown in Table 3.

[0054] Table 3 TCPTP inhibitory activity assay

[0055]

[0056] When the compound concentration is 20 μg / mL, if the inhibition rate is around 50%, the test will continue at the 5 μg / mL concentration level.

[0057] The test results show that compound E exhibits a significant inhibitory effect on non-receptor tyrosine-specific phosphatases and has good clinical application prospects in lowering blood sugar and anti-tumor effects.

[0058] The activity of TCPTP, a phosphatase in the hypothalamus, is closely related to fat metabolism: when fasting, TCPTP is activated, thereby inhibiting the insulin signal of the brain's AgRP / NPY neurons, preventing the "browning" of white fat, and reducing energy consumption. Conversely, food intake reduces the activity of TCPTP enzymes in the hypothalamus, thereby upregulating insulin signals, promoting fat browning, and accelerating energy consumption.

[0059] Insulin can regulate glucose metabolism in peripheral tissues and the brain. The insulin receptor signaling pathway can inhibit the AgRP neurons located in the hypothalamus to promote insulin's inhibition of glycogen synthesis in the periphery, and the activation of AgRP neurons will weaken the glucose uptake capacity of brown adipose tissue. Previous studies have found that the tyrosine phosphatase TCPTP can inhibit the insulin receptor signaling of AgRP neurons. TCPTP in the hypothalamus will be induced to express under hunger conditions, while TCPTP will be degraded after eating. TCPTP can control the insulin receptor signaling of AgRP neurons, coordinate glycogen synthesis and glucose uptake of brown / beige adipocytes in response to eating and hunger.

[0060] The above are only preferred embodiments of the present invention, and the protection scope of the present invention is not limited to the above embodiments. All technical solutions under the concept of the present invention belong to the protection scope of the present invention. It should be pointed out that for ordinary technicians in this technical field, any modification, equivalent replacement or improvement without departing from the principle of the present invention should be regarded as the protection scope of the present invention.

Claims

1. A compound for targeted inhibition of TCPTP, characterized in that, The compound structural formula is as follows: In the formula, R is selected from one or more of F, Cl, Br, cyano, alkyl and alkoxy.

2. The compound for inhibiting TCPTP according to claim 1, characterized in that: The compound is selected from one of the following structural formulas; 3. Use of the compound according to claim 1 or 2 in the preparation of a TCPTP inhibitor.

4. Use of the compound according to claim 1 or 2 in the preparation of anti-tumor drugs.

5. Use of the compound according to claim 1 or 2 in the preparation of hypoglycemic drugs.

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