HnRNP A2B1 inhibitor based on indole-1, 2, 4-oxadiazole structure and application of hnRNP A2B1 inhibitor

Abstract

The invention relates to an hnRNP A2B1 inhibitor based on an indole-1, 2, 4-oxadiazole structure and application of the hnRNP A2B1 inhibitor, and belongs to the field of chemical medicines. The structure of the hnRNP A2B1 inhibitor is as shown in a general formula I in the specification; wherein R represents a benzyl group and a derivative group thereof, a tetrahydropyranyl group, a cyclopentenyl group, a furyl group, a cyclohexyl group and a cyclobutyl group. Through precise targeting design, direct inhibition of hnRNP A2B1 (IC50 is as low as 4.5 [mu] M) is realized, the action target is single, the off-target risk of TAK-981 and the clinical transformation bottleneck of the RNAi technology are solved, and a brand new structural framework and experimental data support are provided for research and development of the target medicine.

Description

TECHNICAL FIELD

[0001] The present application relates to an hnRNP A2B1 inhibitor based on an indole-1,2,4-oxadiazole structure and its application, belonging to the field of chemical industry and medicine. BACKGROUND

[0002] Heterogeneous nuclear ribonucleoprotein A2B1 (hnRNP A2B1) is an important DNA / RNA binding protein and a core member of the hnRNP protein family. Through its RNA recognition motif (RRM) and glycine-rich domain, it is widely involved in key biological processes such as transcriptional regulation of gene expression, pre-mRNA splicing, mRNA transport, stability maintenance, and translational regulation. Under normal physiological conditions, the dynamic balance of hnRNP A2B1 between the nucleus and cytoplasm is crucial for its function. However, in recent years, more and more studies have shown that hnRNP A2B1 is abnormally highly expressed, its activity is enhanced, or it is abnormally localized in various disease states, and is closely related to the development of the disease.

[0003] Existing technologies have confirmed that hnRNP A2B1 is significantly highly expressed in various solid tumors and hematological malignancies, including but not limited to cervical cancer, hepatocellular carcinoma, pulmonary arterial hypertension, colorectal cancer, breast cancer, glioblastoma, pancreatic cancer, renal cell carcinoma, neuroblastoma, gastric cancer, and non-small cell lung cancer. Its abnormal expression is positively correlated with tumor cell proliferation, invasion and metastasis, chemotherapy resistance, and poor prognosis. In view of the abnormal expression of hnRNP A2B1, existing technologies mainly adopt RNA interference (RNAi) strategies, including small interfering RNA (siRNA) and short hairpin RNA (shRNA). Although laboratory studies have confirmed that knocking down hnRNPA2B1 has significant therapeutic effects in various disease models, RNAi technology has the following inherent defects, such as low delivery efficiency, poor in vivo stability, potential off-target effects, and immunogenicity, which seriously limit its clinical translation application. In the aspect of small molecule inhibitors, existing technologies are still in the early stages. TAK-981, as a SUMOylation inhibitor, can indirectly affect the function of hnRNP A2B1 by inhibiting its SUMOylation modification, and shows anti-tumor activity in a glioblastoma model.

[0004] However, this compound is not a specific inhibitor of hnRNP A2B1, and its target is wide, which has the risk of off-target toxicity. So far, there is no small molecule inhibitor that directly acts on hnRNP A2B1 with high selectivity and good drugability entering the clinical research stage. SUMMARY

[0005] In view of the problem of the development of small molecule inhibitors acting on hnRNP A2B1 at the present stage, the application provides an hnRNP A2B1 inhibitor based on an indole-1,2,4-oxadiazole structure, which realizes direct inhibition of hnRNPA2B1 (IC50 as low as 4.5 muM) through precise targeting design, and has a single target point, solves the off-target risk of TAK-981 and the clinical conversion bottleneck of RNAi technology, and provides a novel structure skeleton and experimental data support for the development of drugs for the target point.

[0006] The technical scheme of the application is as follows: In a first aspect, the application provides an hnRNP A2B1 inhibitor based on an indole-1,2,4-oxadiazole structure, and the structure of the hnRNP A2B1 inhibitor is shown in the general formula I: ; In the formula, R represents a benzyl group and a derivative thereof, a tetrahydropyranyl group, a cyclopentenyl group, a furanyl group, a cyclohexyl group, and a cyclobutyl group.

[0007] As a specific embodiment of the application, the pharmaceutically acceptable salt of the hnRNP A2B1 inhibitor is also included.

[0008] As a specific embodiment of the application, the benzyl group and the derivative thereof are selected from a phenyl group, a 2-methylphenyl group, a 4-methylphenyl group, a 4-ethynylphenyl group, a 4-fluorophenyl group, a 3-fluorophenyl group, a 2-fluorophenyl group, a 4-hydroxyphenyl group, a 4-methoxyphenyl group, a 2-methoxyphenyl group, and a 4-methylthiophenyl group.

[0009] As a specific embodiment of the application, the pharmaceutically acceptable salt is selected from a hydrochloride, a sulfate, a phosphate, a hydrobromide, a maleate, a citrate, a succinate, a methanesulfonate, and a toluenesulfonate.

[0010] As a specific embodiment of the application, the hnRNP A2B1 inhibitor includes at least one of the following 17 compounds: .

[0011] In a second aspect, the application provides the use of the above-mentioned hnRNP A2B1 inhibitor based on an indole-1,2,4-oxadiazole structure in a drug for preventing and / or treating tumors.

[0012] As a specific embodiment of the application, the tumor includes at least one of cervical cancer, lung cancer, colorectal cancer, breast cancer, glioblastoma, pancreatic cancer, renal cell carcinoma, neuroblastoma, gastric cancer, and non-small cell lung cancer.

[0013] In a third aspect, the present application further provides a pharmaceutical composition for preventing tumors, comprising the above-mentioned hnRNP A2B1 inhibitor based on an indole-1,2,4-oxadiazole structure or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0014] In a fourth aspect, the present application further provides a pharmaceutical composition for treating tumors, comprising the above-mentioned hnRNP A2B1 inhibitor based on an indole-1,2,4-oxadiazole structure or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0015] As a specific embodiment of the present application, the pharmaceutical composition is a preparation, preferably, the preparation comprises tablets, capsules, powders, syrups, liquids, suspensions or injections.

[0016] Advantages The compound of the present application has the inherent advantages of small molecule compounds (strong cell membrane penetration and high metabolic stability). Compared with the delivery difficulties of RNAi technology and the off-target risk of TAK-981, such compounds are more likely to achieve efficient targeted therapy in clinical transformation, and are expected to become the core candidate drug for treating tumors with high expression of hnRNP A2B1. Through precise targeted design, direct inhibition of hnRNP A2B1 (IC50 as low as 4.5 μM) is achieved, and the target is single, solving the off-target risk of TAK-981 and the clinical transformation bottleneck of RNAi technology, providing a new structural skeleton and experimental data support for the development of drugs targeting this site. It lays a solid foundation for subsequent pharmacokinetics, toxicology research and clinical trials, and accelerates the transformation process from laboratory research to clinical application.

[0017] According to the AlphaScreen detection data of Example 2 (Table 2), of the 17 compounds of the present application, 14 showed clear hnRNP A2B1 inhibitory activity (IC50 < 50 μM), of which Formula I-3 (IC50 = 4.5 μM), Formula I-15 (IC50 = 5.6 μM), and Formula I-7 (IC50 = 6.7 μM) had the most significant inhibitory activity, proving that such compounds can efficiently block the binding function of hnRNP A2B1 and RNA, with IC50 values of the compounds distributed between 4.5 μM and > 100 μM, forming a gradient activity library. High-activity compounds (such as Formula I-3, I-7, and I-15) can be directly developed as clinical candidate drugs, and low-to-moderate-activity compounds (such as Formula I-13, IC50 = 24.6 μM; and Formula I-14, IC50 = 18.7 μM) can be used for combination therapy or prodrug design, meeting the needs of different tumor treatment scenarios (such as high-invasiveness tumors and postoperative adjuvant therapy).

[0018] The compound of the present application can block the core pathway of tumor progression from the molecular mechanism by directly inhibiting hnRNP A2B1 activity (IC50 as low as 4.5 μM). In combination with the inhibitory activity of the compound in the examples, it can be inferred that such inhibitors have potential therapeutic effects on the above-mentioned multiple cancers, especially for glioblastoma, pancreatic cancer and the like which have limited existing treatment means, thereby providing a new targeted treatment scheme. DETAILED DESCRIPTION

[0019] The present application will be further described in detail below in conjunction with specific examples. These examples are only used to illustrate the present application and are not used to limit the scope of the present application.

[0020] Example 1: Preparation of compounds 1-17 of general formula I

[0021] The starting material 2 (175 mg, 1 mmol) and the corresponding starting material 1 (1 mmol) were dissolved in DMF (2 mL), and EDCI (1.2 mmol), HOBT (1 mmol) and DIEA (2 mmol) were added, and then reacted under microwave conditions (200 W, 180°C) for 20 minutes. The solvent was concentrated under reduced pressure to obtain the crude product, which was further purified by flash column chromatography to obtain compounds I-1-I-17.

[0022] Table 1. Structure of starting material 1 and spectral data of target compounds I-1-I-17

[0023]

[0024]

[0025]

[0026] Example 2: Test of hnRNP A2B1 inhibitory activity The hnRNP A2B1 inhibitory activity of the compound was tested by AlphaScreen detection method according to the method of reference (Nucleic Acids Research, 2025, 53, gkaf176). The specific method is as follows: in a total volume of 20 microliters of PBS system, the concentration of RNA EXO motif (selected below the hook point) is 17 nanomoles, the concentration of GST-hnRNPA2B1 is 30 nanomoles, and the concentration of the test compound is different. After incubation for 15 minutes, the RNA probe was added and incubated for 60, and the Alpha signal was detected by Enspire plate reader instrument (PerkinElmer), and the IC 50The results are shown in Table 1.

[0027] Example 5: Test of anti-tumor activity in vitro The MTT method was used to test the proliferation inhibition activity of the compounds on tumor cells. The method is as follows: Hela and MCF-7 tumor cells were inoculated in a 96-well plate at a density of 1x10 5 cells / mL, and placed in a 5% CO2 incubator at 37 o C overnight. Six concentration gradients were set for each compound, and three replicates were set for each concentration. Each concentration was added to the corresponding well, and incubated in a 5% CO2, 37 o C incubator for 24 h. MTT (final concentration 0.5 mg / mL) was added, and the change in absorbance value was detected at 490 nm after 3 h of incubation. The IC 50 value of the compound was calculated, and the inhibition activity of the compound on tumor cell proliferation was evaluated. The experimental results are shown in Table 1.

[0028] Table 1: Test results of hnRNP A2B1 inhibition activity and anti-tumor activity of the compounds

[0029] The preliminary research results show that the compounds have obvious hnRNP A2B1 inhibition activity, and have broad application prospects in the preparation of tumor treatment drugs. The series of compounds described in the present application can be used as active ingredients to add conventional pharmaceutical adjuvants to prepare a neurodegenerative disease treatment drug, which can be prepared into any one of the following dosage forms: tablets, capsules, powders, syrups, liquids, suspensions, or injections.

[0030] The above illustrates the preparation steps, identification process, and pharmacological experiment process of the compounds of the present application with specific examples, but those skilled in the art can make various modifications and changes, and the appended claims cover all such modifications within the scope of the present application without departing from the spirit and scope of the present application.

Claims

1. A hnRNP A2B1 inhibitor based on the indole-1,2,4-oxadiazole structure, characterized in that, The structure of the hnRNPA2B1 inhibitor is shown in general formula I: ； Where R represents benzyl and its derivatives, tetrahydropyranyl, cyclopentenyl, furanyl, cyclohexyl, and cyclobutyl.

2. The hnRNP A2B1 inhibitor based on the indole-1,2,4-oxadiazole structure according to claim 1, characterized in that, It also includes pharmaceutically acceptable salts of hnRNP A2B1 inhibitors.

3. The hnRNP A2B1 inhibitor based on the indole-1,2,4-oxadiazole structure according to claim 1, characterized in that, The benzyl group and its derivatives are selected from phenyl, 2-methylphenyl, 4-methylphenyl, 4-ethynylphenyl, 4-fluorophenyl, 3-fluorophenyl, 2-fluorophenyl, 4-hydroxyphenyl, 4-methoxyphenyl, 2-methoxyphenyl, and 4-methylthiophenyl.

4. The hnRNP A2B1 inhibitor based on the indole-1,2,4-oxadiazole structure according to claim 2, characterized in that, The pharmaceutically acceptable salt is selected from hydrochloride, sulfate, phosphate, hydrobromide, maleate, citrate, succinate, methanesulfonate, and toluenesulfonate.

5. The hnRNP A2B1 inhibitor based on the indole-1,2,4-oxadiazole structure according to any one of claims 1-4, characterized in that, The hnRNP A2B1 inhibitor includes at least one of the following 17 compounds; 。 6. The use of the hnRNP A2B1 inhibitor based on the indole-1,2,4-oxadiazole structure according to any one of claims 1-5 in the prevention and / or treatment of tumors.

7. The application according to claim 6, characterized in that, The tumors mentioned include at least one of cervical cancer, lung cancer, colorectal cancer, breast cancer, glioblastoma, pancreatic cancer, renal cell carcinoma, neuroblastoma, gastric cancer, and non-small cell lung cancer.

8. A pharmaceutical composition for preventing tumors, characterized in that, The pharmaceutical composition comprises any one of the indole-1,2,4-oxadiazole-based hnRNP A2B1 inhibitors or pharmaceutically acceptable salts thereof, as well as pharmaceutically acceptable excipients, according to any one of claims 1-5.

9. A pharmaceutical composition for treating tumors, characterized in that, The pharmaceutical composition comprises any one of the indole-1,2,4-oxadiazole-based hnRNP A2B1 inhibitors or pharmaceutically acceptable salts thereof, as well as pharmaceutically acceptable excipients, according to any one of claims 1-5.

10. The tumor drug composition according to claim 8 or 9, characterized in that, The pharmaceutical composition is a formulation, preferably comprising tablets, capsules, powders, syrups, liquids, suspensions, or injections.

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