Arginine methyltransferase prmt3 inhibitor and use thereof

Abstract

The application discloses a kind of targeting arginine methyltransferase PRMT3 inhibitor and its application, and its effective component is WL43 and its pharmaceutically acceptable salt, and the WL43 is 2-(benzo[c][1,2,5]oxadiazole-5-carbonyl)-N-(3,5-dichlorophenyl) hydrazine carboxamide.The molecular formula of WL43 in the application is C 14 H9Cl2N5O3, has obvious PRMT3 methylation inhibition activity, can significantly inhibit cancer cell growth proliferation, so as to play the role of treating cancer.

Description

Technical Field

[0001] This invention belongs to the field of anticancer drug technology, specifically relating to an inhibitor targeting arginine methyltransferase PRMT3 and its application. Background Technology

[0002] Cancer is one of the leading causes of death worldwide, and its incidence rate is increasing year by year, with the affected population showing a trend towards younger ages. Therefore, identifying regulatory factors that play a key role in the development and progression of cancer and designing effective and specific small molecule inhibitors based on these factors is of significant academic value and social benefit.

[0003] Protein methylation is one of the most important forms of epigenetic regulation and a crucial post-translational modification. The arginine methyltransferase (PRMT) family is a vital component of protein methyltransferases. Based on their different catalytic mechanisms, PRMTs are divided into three subfamilies: ① Type I arginine methyltransferases, which catalyze both monomethylation and asymmetric dimethylation of arginine, including PRMT1, PRMT3, PRMT4 / PRMT3, PRMT6, and PRMT8; ② Type II arginine methyltransferases, which catalyze both monomethylation and symmetric dimethylation of arginine, including PRMT5 and PRMT9; and ③ Type III arginine methyltransferase, PRMT7, which catalyzes only monomethylation of arginine. Protein arginine methylation is a post-translational modification catalyzed by protein arginine methyltransferases (PRMTs) and participates in various physiological processes, such as regulating gene expression, DNA repair, and RNA splicing. Studies have found that abnormal expression and dysfunction of protein methyltransferases (PMTs) are closely related to the occurrence and development of cancer. Research indicates that members of the PRMTs arginine methyltransferase family, through epigenetic regulatory mechanisms, participate in various biological processes and are closely related to cancer development.

[0004] Protein arginine methyltransferase 3 (PRMT3) is a type I PRMT that plays important physiological roles in regulating gene expression, lipid metabolism, and bone development. In 1998, PRMT3 was first discovered as a cytoplasmic protein that methylates the 40S ribosomal protein S2 (rpS2), affecting the normal maturation of the 80S ribosome. Studies have found that abnormal PRMT3 methylation can promote the development and progression of many cancers; for example, PRMT3 overexpression in colorectal cancer promotes cell proliferation, migration, and invasion by stabilizing c-MYC and HIF1α. Furthermore, PRMT3 promotes glycolysis and affects hepatocellular carcinoma growth by enhancing arginine methylation of lactate dehydrogenase A. On the other hand, PRMT3 can not only affect the development and progression of pancreatic cancer through metabolic reprogramming, but also increase ABCG2 expression by methylating hnRNPA1, thereby improving chemotherapy resistance in pancreatic cancer. Furthermore, researchers have found that dysmethylation of the PRMT3 protein is closely related to breast cancer, endometrial cancer, invasive microcarcinoma of the breast, and glioblastoma.

[0005] In summary, PRMT3, as an epigenetic regulator, possesses multiple biological functions. Functional studies of arginine methylation in cancer indicate that this epigenetic regulator is a potential target for anti-tumor drugs, and currently, no inhibitors targeting PRMT3 have entered clinical trials. Therefore, in-depth research into the application of PRMT3 inhibitors in cancer, and the development of targeted drugs to meet diverse clinical needs, is of great significance for cancer treatment. Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of the prior art and provide an inhibitor that targets arginine methyltransferase PRMT3.

[0007] Another object of the present invention is to provide the use of the above-mentioned arginine methyltransferase PRMT3 inhibitor in the preparation of cancer therapeutic compositions.

[0008] Another object of the present invention is to provide a cancer treatment composition.

[0009] The technical solution of the present invention is as follows:

[0010] A PRMT3 inhibitor targeting arginine methyltransferase, the active ingredient of which is WL43 and its pharmaceutically acceptable salt, wherein WL43 is 2-(benzo[c][1,2,5]oxadiazole-5-carbonyl)-N-(3,5-dichlorophenyl)hydrazine carboxamide, and its structural formula is as follows:

[0011]

[0012] In a preferred embodiment of the invention, the pharmaceutically acceptable salt is a salt formed by 2-(benzo[c][1,2,5]oxadiazole-5-carbonyl)-N-(3,5-dichlorophenyl)hydrazine carboxamide with an acid and / or a salt formed with an acidic salt of an inorganic base.

[0013] More preferably, the acid is hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-benzenesulfonic 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, and the acidic salt of the inorganic base is a salt containing a basic metal cation, a salt containing an alkaline earth metal cation, or a salt containing ammonium ions.

[0014] The use of the above-mentioned arginine methyltransferase PRMT3 inhibitor in the preparation of cancer therapeutic compositions.

[0015] In a preferred embodiment of the invention, the cancer treatment composition further includes pharmaceutically acceptable excipients.

[0016] In a preferred embodiment of the present invention, the cancer includes colorectal cancer, breast cancer, liver cancer, pancreatic cancer, glioblastoma, and endometrial cancer.

[0017] A cancer treatment composition comprising the above-mentioned PRMT3 inhibitor targeting arginine methyltransferase.

[0018] In a preferred embodiment of the present invention, the active ingredient is the PRMT3 inhibitor targeting arginine methyltransferase.

[0019] In a preferred embodiment of the invention, the cancer treatment composition further includes pharmaceutically acceptable excipients.

[0020] More preferably, the cancers include colorectal cancer, breast cancer, liver cancer, pancreatic cancer, glioblastoma, and endometrial cancer.

[0021] The beneficial effects of this invention are: the molecular formula of 2-(benzo[c][1,2,5]oxadiazole-5-carbonyl)-N-(3,5-dichlorophenyl)hydrazine carboxamide in this invention is C 14 H9Cl2N5O3 exhibits significant PRMT3 methylation inhibitory activity, which can significantly inhibit the growth and proliferation of cancer cells, thereby playing a role in cancer treatment. Detailed Implementation

[0022] The technical solution of the present invention will be further explained and described below through specific embodiments.

[0023] In the following examples, WL43 is 2-(benzo[c][1,2,5]oxadiazole-5-carbonyl)-N-(3,5-dichlorophenyl)hydrazine carboxamide, with the following structural formula:

[0024]

[0025] Its molecular formula is C 14 H9Cl2N5O3 was purchased from the Maybridge molecular library, Maybridge compound number KM09657.

[0026] Example 1

[0027] The methylation reaction was performed using the MTase-Glo Methyltransferase Assay kit from Promega. The methylation reaction system consisted of: 4×reaction buffer (80mM Tris-HCl 8.0, 200mM NaCl, 4mM EDTA, 12mM MgCl2, 0.4mg / mL BSA), 20μM SAM, 25μM histone substrate, 0.5ng PRMT3, and DMSO solution with a WL43 concentration gradient. After adding MTase-Glo Reagent, the methylation reaction was carried out at 37℃ for 2-3 hours. After the methylation reaction was complete, MTase-Glo Detection Solution was added, and the reaction was continued at 37℃ for 30 minutes. The mixture was then transferred to a 96-well plate, and the fluorescence intensity was measured using the GloMax Discover System. In this embodiment, no WL43 was added to the negative control group system, and no methyltransferase or WL43 was added to the blank control group system. The concentrations of WL43 in the experimental groups were 0.01 μM, 0.02 μM, 0.04 μM, 0.08 μM, 0.16 μM, 0.32 μM, 0.62 μM, 1.25 μM, 2.50 μM, 5.00 μM, 10.00 μM, 20.00 μM, 40.00 μM, 60.00 μM, 80.00 μM, 100.00 μM, and 200.00 μM, respectively. The fluorescence intensity values ​​of the experimental and control groups at the concentration gradients were analyzed using GraphPad Prism 9.0 software, and the enzyme inhibitory activity was calculated (the half-inhibition concentration IC50 was used in this embodiment). 50 (Indicated). The enzyme inhibitory activity data detected in each experiment are shown in Tables 1 and 2.

[0028] Table 1. Enzyme inhibitory activity of WIA3

[0029] compound <![CDATA[IC 50 (micromolar, μM) WL43 93.34

[0030] Table 2 Enzyme inhibitory activity of WIA3

[0031]

[0032]

[0033] As shown in Tables 1 and 2, WL43 exhibits significant inhibitory activity against PRMT3 methylation, with a low half-maximal inhibitory concentration (HMC). Since PRMT3 plays a crucial role in cancer cell growth and proliferation, WL43 can be used in drugs for the prevention or treatment of diseases associated with PRMT3 inhibitors, particularly in related cancer treatments.

[0034] Example 2

[0035] In this embodiment, cells from different breast cancer cell lines (MCF7, HCC1806, and HCC1937) were seeded in 96-well plates at a density of 20%–30%. After 24 hours, DMSO solutions containing different concentrations of WL43 were added. Twelve concentration gradients of the test drug (WL43) were established: 400.00 μM, 200.00 μM, 100.00 μM, 80.00 μM, 60.00 μM, 40.00 μM, 20.00 μM, 10.00 μM, 5.00 μM, 2.50 μM, 1.00 μM, and 0.50 μM. Each concentration gradient included three replicates.

[0036] Change the cell culture medium before drug administration. 48 hours after drug administration, use (CellTiter) Aqueous (single-solution cell proliferation assay kit) detects cytotoxicity using a colorimetric method. CellTiter AQueous single-solution reagent contains a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt; MTS] and an electron coupling reagent (ethylthiophenazine, PES).

[0037] Add 20 μL of CellTiter per 100 μL of culture medium. The AQueous solution was incubated at 37°C in a 5% CO2 incubator for 1 hour. The reaction was terminated by adding 25 μL of 10% SDS.

[0038] This embodiment also includes another comparative experiment. All reaction conditions are consistent with those described above, except that WL43 is replaced with DMSO solution of the existing PRMT3 enzyme activity inhibitor SGC707 (purchased from MCE, catalog number HY-19715) as a positive control, and only DMSO solution is added to the blank control. Absorbance data at 490 nm were recorded using a Thermo Multiskan MK3 microplate reader. The data were processed, and the IC50 of the drug was analyzed using GraphPad Prism 9.0 software. 50 Values. The test results are shown in Table 3.

[0039] Table 3. Cell proliferation inhibition data

[0040] <![CDATA[IC 50 ]]> WL43(μM) SGC707 (μM) Blank group MCF7 41.85 40.85 - HCC1806 54.56 70.84 - HCC1937 56.10 153.00 -

[0041] As shown in Table 3, WL43 has a significant technical effect in inhibiting the proliferation of breast cancer cells, and it can effectively improve the efficiency of inhibiting the proliferation of breast cancer cells compared with SGC707. WL43 can be used in drugs for the prevention or treatment of diseases related to PRMT3 inhibitors, especially in related cancer treatment drugs.

[0042] The above description is merely a preferred embodiment of the present invention, and therefore should not be construed as limiting the scope of the present invention. All equivalent changes and modifications made in accordance with the scope of the patent and the contents of the specification should still fall within the scope of the present invention.

Claims

1. The use of a PRMT3 inhibitor targeting arginine methyltransferase in the preparation of a cancer therapeutic composition, characterized in that: The active ingredient of this PRMT3 inhibitor targeting arginine methyltransferase is WL43 and its pharmaceutically acceptable salt. WL43 is 2-(benzo[c][1,2,5]oxadiazole-5-carbonyl)-N-(3,5-dichlorophenyl)hydrazine carboxamide, with the following structural formula: , The cancer is breast cancer.

2. The use as described in claim 1, characterized in that: The cancer treatment composition also includes pharmaceutically acceptable excipients.

3. The use as described in claim 1, characterized in that: The pharmaceutically acceptable salt is a salt formed by 2-(benzo[c][1,2,5]oxadiazole-5-carbonyl)-N-(3,5-dichlorophenyl)hydrazine carboxamide with an acid and / or with an acidic salt of an inorganic base.

4. The use as described in claim 3, characterized in that: The acid is hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-benzenesulfonic 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, and the acidic salt of the inorganic base is a salt containing a basic metal cation, a salt containing an alkaline earth metal cation, or a salt containing ammonium ions.

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