Application of arbidol in preparation of antitumor drugs
By using the antiviral drug Abidol to inhibit PKM2 and regulate tumor metabolism and immune response, the target specificity and toxicity of existing targeted sugar metabolism drugs have been solved, and broad-spectrum anti-tumor effects and enhancement of immunotherapy have been achieved.
Patent Information
- Application Number
- CN202510703235.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-29
- Publication Date
- 2025-08-01
AI Technical Summary
Existing targeted sugar metabolism drugs have insufficient target specificity and toxicity problems in clinical applications, resulting in forced termination in the early stages and lack of effective anti-tumor strategies. In particular, there has been no breakthrough in the development of drug targeting PKM2.
Umifenovir, a marketed antiviral drug, was used as a PKM2 inhibitor, to regulate tumor metabolism reprogramming by inhibiting the dual enzyme activity of PKM2, combining immunomodulation, and enhancing the effects of chemotherapy and immunotherapy.
Abidol can significantly inhibit the growth of a variety of tumor cells, improve chemotherapy resistance, improve immunotherapy effects, reduce R&D risks and costs, and provide broad-spectrum anti-tumor effects.
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Figure CN120392746A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of biopharmaceutical technology, and particularly to the application of Arbidol in the preparation of anti-tumor drugs. Background Art
[0002] In recent years, abnormal energy metabolism, as one of the ten hallmarks of tumors, the research and development of anti-tumor drugs that selectively regulate tumor metabolic reprogramming has become a research hotspot in the field of oncology. However, there are still many bottlenecks in the process from basic research to clinical translation. At present, many scientists are committed to inhibiting tumor proliferation by targeting key enzymes (such as HK, PFK, PK, LDH, etc.) or glucose transporters (GLUT) in the glycolytic pathway. Although these candidate drugs targeting glucose metabolism have shown significant effects in preclinical studies, due to insufficient target specificity, most drugs are forced to be terminated in the early clinical stage due to toxicity problems or insignificant therapeutic benefits. In particular, the problem of toxicity control has become the main obstacle restricting their clinical application. After years of exploration, there is still no breakthrough in the clinical research of drugs targeting tumor energy metabolism. This situation highlights the technical bottleneck in the development of metabolic reprogramming-targeted drugs and also indicates that the anti-tumor strategy targeting metabolism is facing severe challenges.
[0003] In the complex regulatory network of energy metabolism, pyruvate kinase M2 isoform (PKM2) has gradually stood out from numerous metabolic pathways and enzymes due to its unique biological characteristics and targeted therapeutic potential, and has been proven to be one of the therapeutic targets with high selectivity and translational research value. Pyruvate kinase is the last rate-limiting enzyme in the glycolytic pathway, which can irreversibly catalyze phosphoenolpyruvate (PEP) and adenosine diphosphate (ADP) to generate pyruvate and adenosine triphosphate (ATP). It is worth noting that PKM2 is highly specifically expressed in rapidly proliferating tissues and tumor cells, in sharp contrast to other isoforms (PKM1, PKR, and PKL) mainly present in mature tissues. This significant difference between tumor cells and normal cells makes PKM2 an ideal target for tumor metabolic intervention, providing an important basis for the development of highly selective anti-tumor drugs. Different from the PKM1 isoform that can spontaneously form tetramers, PKM2 has a unique "dimer / tetramer" structural conversion property. Among them, the stable tetramer form has high glycolytic activity, and the dimer form has low glycolytic activity. This dynamic structural change enables it to precisely regulate energy metabolism and anabolic metabolism in tumor cells to meet the needs of rapid tumor proliferation.
[0004] More importantly, PKM2 not only has the catalytic function of the rate-limiting enzyme of glycolysis, but also can enter the nucleus in the dimer state, showing its unique protein kinase activity. By phosphorylating key transcription factors (such as P38, HIF-1α, STAT3, etc.), histones and other signaling molecules, PKM2 directly participates in the regulation of multiple gene transcriptions and signaling pathways. In tumor cells, PKM2 participates in the regulation of multiple genes through its protein kinase activity, thus playing a core role in the processes of tumor proliferation, invasion and metastasis. In immune cells in the tumor microenvironment, PKM2 can also regulate the expression of various cytokines (such as IL-1β, IL-10, etc.) and immune-related genes such as PD-L1 through transcription factors and signaling pathways such as P38, ERK1 / 2, NF-κB, STAT3, etc. Another literature report shows that knocking out PKM2 can reshape the functional state of immune cells, thereby improving the efficacy of immunotherapy. This dual property of metabolic regulation and transcriptional regulation, as well as its multi-dimensional role in tumor cells and the immune microenvironment, make PKM2 a highly potential multi-functional target in tumor treatment, providing an important theoretical basis for the development of new anti-tumor strategies.
[0005] At present, the research and development of small molecule drugs targeting PKM2 is still in the early exploration stage. Only two candidate drugs, TP-1454 and ACT-001, have entered clinical research, and are in the phase I and phase II clinical trial stages respectively. Both of these drugs act as PKM2 tetramer stabilizers, regulating tumor metabolic reprogramming by enhancing the tetramer form of PKM2. Although the preliminary research data shows its potential value in inhibiting tumor growth and improving treatment resistance, due to the multi-stage verification process, long-term efficacy evaluation and safety monitoring involved in clinical trials, these drugs still have a long R & D cycle before actual clinical application. Summary of the Invention
[0006] The object of the present invention is to provide the application of Arbidol in the preparation of anti-tumor drugs.
[0007] In order to achieve the above-mentioned invention object, the present invention provides the following technical solutions:
[0008] The present invention provides the application of Arbidol in the preparation of anti-tumor drugs.
[0009] Preferably, the anti-tumor drug is a drug for treating the following cancers: lung cancer (small cell lung cancer, lung adenocarcinoma), cervical cancer, ovarian cancer, gastric cancer, breast cancer (triple negative type, HR positive type, HER positive type), prostate cancer, bladder cancer, bone cancer, liver cancer, pancreatic cancer, esophageal cancer, colon cancer.
[0010] Preferably, the anti-tumor drug is a tumor cell inhibitor.
[0011] Preferably, the tumor cell inhibitor is a pharmaceutical preparation that inhibits the proliferation of tumor cells.
[0012] Preferably, the tumor cell inhibitor is a pharmaceutical preparation that inhibits the growth of tumor cells. Preferably, the tumor cell inhibitor is a tumor cell killing and eliminating preparation.
[0013] Preferably, umifenovir can regulate tumor metabolic reprogramming by inhibiting the activity of PKM2 pyruvate kinase, playing a role in directly killing tumors.
[0014] The present invention also provides a drug for treating cancer, comprising umifenovir and a pharmaceutically acceptable excipient.
[0015] Preferably, the cancer is lung cancer (small cell lung cancer, lung adenocarcinoma), cervical cancer, ovarian cancer, gastric cancer, breast cancer (triple negative type, HR positive type, HER positive type), prostate cancer, bladder cancer, bone cancer, liver cancer, pancreatic cancer, esophageal cancer, colon cancer.
[0016] Preferably, the dosage form of the drug is capsule, pill, tablet, granule or injection.
[0017] Compared with the prior art, the present invention has the following beneficial effects:
[0018] The present invention uses the previously established SERScreen high-throughput drug screening method to screen out the drug Umifenovir (umifenovir) that can specifically inhibit the dual enzyme activity of PKM2. As an antiviral drug that has been marketed and widely used, the safety of umifenovir has been fully recognized in long-term clinical use and systematic research. With the existing data, the redevelopment of the anti-tumor activity of Umifenovir can simplify some of the previous tests, reduce the R & D risk and time investment, accelerate the clinical transformation speed of new therapies, save a large amount of costs for society, and enable more patients to benefit from it earlier.
[0019] According to the applicant's previous research results, Umifenovir can not only directly kill tumors by regulating metabolic reprogramming, but also enhance the immune response by regulating the activation and proliferation of lymphocytes, and significantly inhibit the expression of a variety of cytokines. Therefore, Umifenovir has important potential value both as a single anti-tumor drug and in improving the efficacy of chemotherapy, improving the inhibitory immune microenvironment, enhancing the effect of immunotherapy, and preventing and treating common adverse reactions of immunotherapy such as cytokine release syndrome.
[0020] As a non-nucleoside broad-spectrum antiviral drug, the broad-spectrum anti-tumor effect of umifenovir has not been reported. Umifenovir can regulate tumor metabolic reprogramming by inhibiting the activity of PKM2 pyruvate kinase, playing a role in directly killing tumors. Previous work has shown that it has good killing effects on 49 tumor cell lines in 8 systems. Umifenovir inhibits the activity of PKM2, the key enzyme at the end of the glycolysis pathway, and can affect DNA damage repair by blocking the generation of cell synthesis raw materials, thereby enhancing the efficacy of chemotherapy and radiotherapy. Umifenovir can inhibit the nuclear entry of PKM2 to exert protein kinase activity, enhance the activity and function of lymphocytes, reshape the immune microenvironment, and enhance the anti-tumor immune efficacy, showing broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS
[0021] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the drawings in the following description are only the embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained according to the provided drawings.
[0022] Figure 1 Is the half-lethal concentration (IC 50 ) of umifenovir against 49 tumor cells in different systems.
[0023] Figure 2 Is the statistical chart of the killing activity of umifenovir against tumor cells.
[0024] Figure 3 Is the statistical chart of the killing activity of umifenovir against tumor cells from different tissue sources.
[0025] Figure 4 Is the predicted diagram of the molecular docking binding mode of umifenovir and PKM2.
[0026] Figure 5 Is the diagram of umifenovir inhibiting the activity of PKM2 pyruvate kinase at different concentrations; A - B: The diagram of umifenovir inhibiting the activity of pyruvate kinase at different concentrations; C: The diagram of umifenovir specifically inhibiting the activity of PKM2; D: The protein sequence difference diagram of PKM1 and PKM2; E: The electrophoresis diagram of umifenovir inhibiting the nuclear translocation of PKM2.
[0027] Figure 6It is the diagram of umifenovir blocking the glycolysis pathway at pyruvate kinase; A: Metabolomics analysis suggests that umifenovir inhibits tumor glycolysis; B: Schematic diagram of pathway and metabolite changes; C: Umifenovir inhibits the levels of key metabolites in the glycolysis pathway.
[0028] Figure 7 It is the diagram of umifenovir blocking the compensatory activation of metabolic bypass in glycolysis; A: Volcano plot of differential gene expression in cells before and after umifenovir treatment; B: Venn diagram of differentially expressed genes by transcriptome sequencing; C: Summary diagram of differentially expressed gene information; D: Heat map of differentially expressed genes.
[0029] Figure 8 It is about umifenovir combined with cisplatin enhancing the tumor killing effect on the platinum-resistant cell line SKOV3-CDDP.
[0030] Figure 9 It is about umifenovir combined with programmed death receptor-1 (PD-1) monoclonal antibody enhancing the efficacy of immunotherapy.
[0031] Figure 10 It is about umifenovir combined with CAR-T therapy enhancing the efficacy of immunotherapy. Specific implementation manners
[0032] The technical solutions provided by the present invention will be described in detail below in conjunction with embodiments, but they should not be construed as limiting the protection scope of the present invention.
[0033] Example 1
[0034] ① Inoculate cells: Inoculate 1×10 4 tumor cells in the logarithmic growth phase into a 96-well plate, and use the corresponding culture medium containing 2% serum.
[0035] ② Drug treatment: Add umifenovir solution with gradient concentrations to a total volume of 0.1 ml while inoculating cells. At the same time, set up a blank control group and a negative control group, with 3 parallel controls in each group. Then place the culture plate in a constant temperature incubator at 37°C and 5% CO2 for 24 h.
[0036] ③ Detection of survival rate: Add CCK-8 reagent to each well, incubate for 4 h, measure the absorbance value of each well at a wavelength of 450 nm using a multifunctional microplate reader, and calculate the cell survival rate through GraphPad Prism software. Calculation formula:
[0037] Cell survival rate (%) = (OD value of experimental group - OD value of blank control group) / (OD value of negative control group - OD value of blank control group) × 100%
[0038] ④ Calculate the median lethal concentration: Use GraphPad Prism software for non - linear regression analysis to obtain the fitted dose - survival rate curve and IC 50 , and quantitatively evaluate the killing effect of Umifenovir on tumor cells.
[0039] At present, there is no report on whether the anti - tumor effect of Umifenovir is extensive. The applicant evaluated the killing effect of Umifenovir on various tumors in multiple systems through in vitro tumor cell killing experiments, and found that Umifenovir has a good killing effect on 49 tumor cell lines of common various systems, and has no obvious killing effect on normal human peripheral blood mononuclear cells (PBMC) ( Figure 1 ). And according to their median lethal concentration (IC 50 ) of tumor cells, they were divided into high killing activity (1 - 15 μM), medium killing activity (15 - 30 μM) and low killing activity (> 30 μM). Analysis found that Umifenovir has strong killing activity against 30.61% of tumor cells and medium or above killing activity against 85.71% of tumor cells ( Figure 2 ), and no drug resistance was shown in tumors from different tissue sources, and all can play a good killing role ( Figure 3 ). These results indicate that the anti - tumor effect of Umifenovir is broad - spectrum.
[0040] Example 2
[0041] (1) Docking prediction analysis of Umifenovir with PKM2 molecule
[0042] To evaluate the affinity of Umifenovir for its target PKM2, we performed molecular docking analysis. Use Autodock Vina v.1.2.2 software to predict their binding mode and interaction, and calculate the binding energy. The results show that Umifenovir can bind to PKM2 ( Figure 4 ), and the binding energy is - 9.61 kcal / mol, indicating that their binding is stable.
[0043] (2) Umifenovir can selectively inhibit the pyruvate kinase activity and nuclear translocation of PKM2
[0044] To confirm whether Umifenovir regulates tumor metabolism by inhibiting pyruvate kinase activity, we detected the pyruvate kinase activity of pancreatic cancer cells under treatment with different concentrations of Umifenovir. The results show that as the concentration of Umifenovir increases, the pyruvate kinase activity continuously decreases ( Figure 5.(A - B) indicates that Umifenovir clearly inhibits the activity of this key rate - limiting enzyme in the glycolysis pathway.
[0045] To further confirm whether Umifenovir selectively inhibits PKM2 activity, we constructed stable BxPC - 3 cell lines overexpressing PKM1 and PKM2, and also detected the activity of pyruvate kinase under Umifenovir treatment. The results showed that Umifenovir could only reduce the PK kinase activity in PKM2 - overexpressing cells, indicating that this inhibitory effect is specific ( Figure 5 .(C). The PKM1 and PKM2 isoforms result from alternative splicing differences in exons 9 and 10, leading to differences in their protein structures and functions ( Figure 5 .(D). In addition, through cytoplasmic / nuclear protein separation analysis, it was found that Umifenovir could significantly inhibit the nuclear translocation of PKM2 ( Figure 5 .(E).
[0046] (3) Metabolomics analysis of Umifenovir - regulated metabolic reprogramming
[0047] To explore the effect of Umifenovir on tumor metabolic reprogramming, we performed metabolomics analysis on pancreatic cancer cells BxPC - 3 and Panc - 1 before and after the treatment in Example 1. The results showed that after Umifenovir treatment, the metabolites upstream of pyruvate kinase in the glycolysis pathway increased, while the metabolites downstream and in the tricarboxylic acid cycle decreased. At the same time, the metabolites in the pentose pathway increased ( Figure 6 ), which suggests that Umifenovir inhibits the glycolysis pathway at pyruvate kinase and activates the pentose phosphate pathway to a certain extent.
[0048] (4) Umifenovir inhibits glycolysis and induces the up - regulation of the metabolic bypass gene UPP1
[0049] To further explore the mechanism of Umifenovir - regulated metabolism, we performed transcriptome sequencing analysis in 5 types of tumor cells with high killing activity. The results showed that the UPP1 gene was significantly up - regulated after Umifenovir treatment. The uridine phosphorylase encoded by it catalyzes the metabolism of uridine in the catalytic cycle to uracil and ribose, and the latter enters the pentose phosphate pathway (PPP), providing energy and nucleotide precursors for the cell. This result indicates that Umifenovir maintains energy homeostasis and biosynthetic requirements by blocking the glycolysis pathway and inducing tumor cells to activate the UPP1 - mediated metabolic bypass ( Figure 7 ).
[0050] Example 3
[0051] 1. Umifenovir combined with cisplatin enhances the antitumor effect on the platinum-resistant cell line SKOV3-CDDP:
[0052] To explore the improvement effect of Umifenovir on tumor chemoresistance, we treated the platinum-resistant ovarian cancer cell line SKOV3-CDDP with cisplatin (CDDP) combined with Umifenovir. The results showed that Umifenovir could significantly enhance the killing effect of cisplatin on the resistant cell line, providing a new potential treatment strategy for overcoming chemoresistance.
[0053] 2. Umifenovir combined with programmed death receptor-1 (PD-1) monoclonal antibody enhances the efficacy of immunotherapy
[0054] To explore the promoting effect of Umifenovir on the efficacy of PD-1 monoclonal antibody, we treated the ovarian cancer cell SKOV3 with Umifenovir combined with pembrolizumab at different effector-to-target ratios (E:T ratios). The results showed that Umifenovir could enhance the regulatory effect of pembrolizumab on T cell killing ability at E:T ratios of 40:1 and 20:1, and its effect increased with the increase in the concentration of Umifenovir.
[0055] 3. Umifenovir combined with CAR-T therapy enhances the efficacy of immunotherapy
[0056] To explore the promoting effect of Umifenovir on the efficacy of chimeric antigen receptor T cell (CAR-T), we treated the ovarian cancer cell SKOV3 with Umifenovir combined with mesothelin-CAR-T cells at different CAR-T cell-to-target ratios. The results showed that Umifenovir could enhance the regulatory effect of pembrolizumab on T cell killing ability at E:T ratios of 40:1, 20:1, 10:1 and 5:1, and its effect increased with the increase in the concentration of Umifenovir.
[0057] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.
Claims
1. Use of Arbidol in the preparation of anti-tumor drugs.
2. The application according to claim 1, characterized in that The anti-tumor drug is a drug for treating the following cancers: lung cancer, cervical cancer, ovarian cancer, gastric cancer, breast cancer, prostate cancer, bladder cancer, bone cancer or liver cancer.
3. The application according to claim 1 or 2, characterized in that, The anti-tumor drug is a tumor cell inhibitor.
4. The application according to claim 3, wherein The tumor cell inhibitor is a pharmaceutical preparation for inhibiting the proliferation of tumor cells.
5. The application according to claim 3, wherein The tumor cell inhibitor is a pharmaceutical preparation for inhibiting the growth of tumor cells.
6. The application according to claim 3, characterized in that, The tumor cell inhibitor is a tumor cell killing preparation.
7. A drug for treating cancer, comprising Arbidol and a pharmaceutically acceptable excipient.
8. The medicament according to claim 7, wherein The cancer is lung cancer, cervical cancer, ovarian cancer, gastric cancer, breast cancer, prostate cancer, bladder cancer, bone cancer or liver cancer.
9. Use of arbidol in combination for preparing an anti-tumor drug, characterized in that, Arbidol is used in combination with cisplatin, PD-1 or CAR-T.
10. The drug according to claim 9, characterized in that, The anti-tumor drug is a drug for treating the following cancers: lung cancer, cervical cancer, ovarian cancer, gastric cancer, breast cancer, prostate cancer, bladder cancer, bone cancer or liver cancer.