Composition and application thereof in tumor anti-lactylation

Abstract

The invention provides a composition and an application thereof in tumor anti-lactylation, and belongs to the technical field of biological medicines. The corilagin and the baicalein are combined to be applied to tumor treatment, a stronger glycolysis inhibition effect can be shown, a main energy supply way of tumor cells can be more effectively cut off, and rapid proliferation and growth of the tumor cells are restrained. The combination of corilagin and baicalein provided by the invention provides a new potential molecular combination drug for lung cancer treatment, provides a new treatment choice for patients, develops a new idea for the field of tumor treatment, proves the potential of a natural compound combination in tumor anti-lactic acid treatment, and has a wide application prospect. Reference and reference are provided for subsequent research and development of more tumor treatment drugs and strategies based on natural products.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to a composition and its application in tumor anti-lactic acidification. Background Art

[0002] Tumor metabolic reprogramming is one of the important characteristics of tumor cells, among which abnormal enhancement of glycolysis plays a key role in the occurrence, development and metastasis of tumors. Even under aerobic conditions, tumor cells preferentially obtain energy through the glycolysis pathway. This phenomenon is called the "Warburg effect", which results in the production of a large amount of lactic acid.

[0003] Excessive tumor lactate levels have a profound impact on the tumor microenvironment. On the one hand, the acidic microenvironment can promote the invasion and metastasis of tumor cells, enhance the migration ability of tumor cells, make it easier for them to break through the basement membrane, invade surrounding tissues and blood vessels, and then metastasize to distant sites. On the other hand, the acidic environment can also inhibit the body's immune response, interfere with the normal function of immune cells such as T cells and NK cells, and enable tumor cells to escape the surveillance and attack of the body's immune system. In addition, enhanced tumor glycolysis provides the necessary energy and biosynthetic precursors for the rapid proliferation of tumor cells, supporting the continuous division and growth of tumor cells.

[0004] Currently, therapeutic strategies targeting tumor glycolysis and lactate metabolism have attracted much attention. Although some existing anti-metabolite drugs can inhibit the metabolic process of tumor cells to a certain extent, they generally have the problem of insufficient specificity. For example, some drugs not only act on tumor cells, but also interfere with the metabolism of normal cells, thereby causing serious adverse reactions, such as bone marrow suppression, gastrointestinal reactions, etc., which reduce the patient's quality of life and tolerance to treatment. In addition, single drug treatment is often difficult to completely block the complex tumor metabolic network. Tumor cells are prone to continue to maintain their abnormal metabolic state through compensatory mechanisms, resulting in limited treatment effects and high tumor recurrence rates. GAPDH and PKM2 are one of the main metabolic enzymes in the glycolysis pathway. Molecular combinations targeting these two targets may have a synergistic effect in reducing lactic acidification.

[0005] Therefore, it is urgent to develop a treatment method with high specificity, low toxicity and side effects that can effectively inhibit tumor glycolysis and reduce tumor lactate levels. Studies have found that natural compounds have great potential in the field of tumor treatment due to their unique chemical structure and biological activity that inhibits glycolysis. Summary of the invention

[0006] The purpose of the present invention is to provide a composition and its application in tumor anti-lactic acidification. The compounds corilagin and baicalein in the present invention are combined to treat tumor anti-lactic acidification, which plays the role of inhibiting tumor glycolysis, reducing tumor lactic acid levels and treating tumors.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions: The invention provides an application of a pharmaceutical composition in preparing a cisplatin treatment synergistic drug; the pharmaceutical composition consists of baicalein and corilagin.

[0008] The present invention also provides a pharmaceutical composition, which comprises baicalein, corilagin and cisplatin.

[0009] The present invention also provides the use of the above-mentioned pharmaceutical composition in the preparation of a cancer cell or cancer organoid proliferation inhibitor.

[0010] The present invention also provides the use of the above-mentioned pharmaceutical composition in preparing drugs for treating cancer.

[0011] The present invention also provides the use of the above-mentioned pharmaceutical composition in the preparation of a cancer cell glycolysis inhibitor.

[0012] The present invention also provides the use of the above-mentioned pharmaceutical composition in preparing a cancer cell lactic acidification inhibitor.

[0013] Preferably, the concentration ratio of baicalein to corilagin is 1:0.8-1.2.

[0014] Preferably, the structural formula of baicalein is as shown in Formula A:

[0015] Formula A The structural formula of the corilagin is shown in Formula B:

[0016] Formula B.

[0017] Preferably, the cancer or cancer cell is selected from one or more of testicular cancer, ovarian cancer, nasopharyngeal cancer, laryngeal cancer, lung cancer, bladder cancer, prostate cancer, gastric cancer or malignant lymphoma.

[0018] Beneficial effects of the present invention: The present invention applies a combination of corilagin and baicalein to tumor treatment. Compared with other related treatment methods or single drugs, the combination of corilagin and baicalein involved in the present invention shows a stronger glycolysis inhibition effect, can more effectively cut off the main energy supply pathway of tumor cells, and curb the rapid proliferation and growth of tumor cells.

[0019] The combination of corilagin and baicalein provided by the present invention provides a new potential molecular combination drug for the treatment of lung cancer, provides patients with a new treatment option, and is expected to improve the treatment effect and prognosis of lung cancer patients. As natural compounds, corilagin and baicalein may have lower toxic side effects than some traditional chemically synthesized antimetabolites. The present invention has opened up new ideas for the field of tumor treatment, demonstrated the potential of natural compound combinations in tumor anti-lactation treatment, and provided reference and reference for the subsequent development of more tumor treatment drugs and strategies based on natural products. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is the result of corilagin binding to GAPDH and inhibiting GAPDH enzyme activity. Figure 1 A is the BLI experimental result of GAPDH binding to different concentrations of corilagin; Figure 1 B shows the inhibition of GAPDH enzyme activity by different concentrations of corilagin; Figure 2 This is the result of baicalein binding to PKM2 and inhibiting PKM2 enzyme activity. Figure 2 A is the BLI experimental results of PKM2 binding to different concentrations of baicalein; Figure 2 B shows the inhibition of PKM2 enzyme activity by different concentrations of baicalin; Figure 3 This is a graph showing the efficacy of the combination of corilagin and baicalin on lung cancer cells. The graph shows the effects of different concentrations of baicalin, corilagin and cisplatin on the formation of A549 cell clones; the units shown in the graph are μM.

[0021] Figure 4 This is the efficacy result of the combination of corilagin and baicalein on lung cancer organoids. Figure 4 A is the inhibitory effect of 10 μM baicalein, 10 μM corilagin, and 2 μM cisplatin on lung cancer organoid 3D spheroids; Figure 4 B is the quantification of the area of ​​lung cancer organoid spheres after co-culture with drugs; Figure 5 This is the result of the combination of corilagin and baicalein reducing lactic acid and lactic acidification in lung cancer cells. Figure 5 A is the content of lactic acid in the cell supernatant measured after 24 h of administration of 10 μM baicalein, 10 μM corilagin, and 2 μM cisplatin, respectively, or in combination; Figure 5 B is the WB of panlactation in cell lysate after 24 h of administration of 10 μM baicalein, 10 μM corilagin, and 2 μM cisplatin, respectively, or in combination; Figure 6 This is a graph showing the efficacy of the combination of corilagin and baicalin on lung cancer-bearing mice. Figure 6A shows the mouse tumors in the control group, cisplatin group, and combined drug administration group from top to bottom; Figure 6 B is the quantitative result of mouse tumor mass; Figure 7 This is a study result of the mechanism of the combination of corilagin and baicalein in reducing lactic acid in lung cancer. Figure 7 A is a CO-IP experiment of MRE11 lactylation; Figure 7 B: CO-IP experiment of AARS1 lactylation DETAILED DESCRIPTION The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0022] Example The efficacy of corilagin combined with baicalin on lung cancer cells 1.1 Target binding and enzyme activity test: The BLI method was used to verify the binding of small molecules to targets. After PKM2 and GAPDH proteins were biotinylated, the SMAP probe was used to load the biotinylated proteins onto the probe. After equilibration in the buffer, they were bound to different concentrations of baicalin and corilagin, respectively, and then eluted. According to the method of the kit, the inhibition of PK enzyme activity by different concentrations of baicalin and the inhibition of GAPDH enzyme activity by different concentrations of corilagin were determined. The results are as follows: Figure 1 and Figure 2 As shown, baicalein and corilagin can bind to PKM2 and GAPDH respectively and effectively inhibit enzyme activity.

[0023] 1.2 Efficacy evaluation: The efficacy of corilagin and baicalin was evaluated using A549 cisplatin-resistant lung cancer cell line and lung cancer organoids. First, cisplatin-resistant A549 cells were seeded into 6-well plates at 2000 cells / well and divided into baicalin (5 μM, 10 μM), corilagin (5 μM, 10 μM), cisplatin (1 μM, 2 μM), baicalin and corilagin, and a combination of the three. After 7-10 days of culture, the clones were stained with crystal violet. The results are shown in Figure 2. Figure 3 Baicalein (10 μM), corilagin (10 μM), cisplatin (2 μM), and the combination of the three effectively inhibited the proliferation of lung cancer organoids after 14 days of administration. Figure 4 These results indicate that baicalein and corilagin can effectively enhance the sensitivity of A549 cisplatin-resistant cells and lung cancer organoids to cisplatin and inhibit the proliferation of lung cancer cells and organoids.

[0024] 1.3 Evaluation of reducing lactic acid: After stimulating the A549 cisplatin-resistant lung cancer cell line with drugs for 24 hours, the lactic acid content in the cell culture medium was determined according to the kit instructions. Figure 5 As shown in A, baicalein and corilagin can effectively reduce the production of lactic acid in cells. After combined administration, the inhibitory effect of lactic acid production can be enhanced. After the cells were washed with PBS, 8M urea containing protease inhibitors was added for protein extraction. Then, the protein was quantified using the BCA kit and the Loading buffer was added to prepare the sample for WB experiment. The results are shown in Figure 5 As shown in B, the combination of baicalein and corilagin can effectively reduce the overall lactic acidification (Pan-Lac) level of cells.

[0025] The efficacy of corilagin combined with baicalin on lung cancer-bearing mice The experimental animals were SPF-grade BALB / c nude mice, 18 males, 5 weeks old, weighing 18-20g. After the A549 cells were cultured and expanded, they were digested with trypsin, washed twice with PBS, and inoculated subcutaneously into the mice. One week later, the mice were randomly divided into a control group, a cisplatin group, and a drug combination group (baicalin, corilagin, and cisplatin). The cisplatin group and the drug combination group were intraperitoneally injected with cisplatin at 3 mg / Kg, once every 3 days. The combination group was intraperitoneally injected with baicalin 10.8 mg / Kg and corilagin 25 mg / Kg, once a day. After 14 days of administration, the mouse tumors were removed and weighed. The results are as follows Figure 6 As shown, the tumor tissue in the combination group was significantly reduced compared with the control group.

[0026] Study on the mechanism of reducing lactic acidosis by the combination of corilagin and baicalein A549 cisplatin-resistant cells were inoculated into culture dishes and administered according to the groups of baicalin (10 μM), corilagin (10 μM), cisplatin (2 μM) and the combination of the three. After 24 h, the protein was extracted using RIPA lysis buffer containing protease inhibitors, phosphatase inhibitors and deacetylase inhibitors, and the protein was quantified using the BCA method. Then, an equal amount of lysate was taken and incubated with agarose beads pre-connected with MRE11 or AARS1 antibodies. After overnight at 4°C, the agarose beads were washed with PBS, and the loading buffer was added and incubated at 95°C for 10 min. After centrifugation, an equal amount of the upper layer sample was taken for SDS-page and transfer, and then blocked with skimmed milk powder and incubated with lactic acid antibody. After incubation with the secondary antibody, the image was developed, and the results are shown as follows. Figure 7 As shown, the combined administration of baicalein, corilagin and cisplatin could reduce the lactic acidification levels of MRE11 and AARSI.

[0027] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A pharmaceutical composition for use in preparing a cisplatin-based synergistic drug; the pharmaceutical composition is composed of baicalein and corilagin.

2. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises baicalein, corilagin and cisplatin.

3. Use of the pharmaceutical composition according to claim 2 in the preparation of a cancer cell or cancer organoid proliferation inhibitor.

4. Use of the pharmaceutical composition according to claim 2 in the preparation of drugs for treating cancer.

5. Use of the pharmaceutical composition according to claim 2 in the preparation of a cancer cell glycolysis inhibitor.

6. Use of the pharmaceutical composition according to claim 2 in the preparation of a cancer cell lactic acidification inhibitor.

7. The pharmaceutical composition according to claim 2, characterized in that The concentration ratio of baicalein to corilagin is 1:0.8-1.

2.

8. The pharmaceutical composition according to claim 2, characterized in that The structural formula of baicalein is shown in Formula A: Formula A The structural formula of the corilagin is shown in Formula B: Formula B.

9. The use according to any one of claims 3 to 8, characterized in that: The cancer or cancer cell is selected from one or more of testicular cancer, ovarian cancer, nasopharyngeal cancer, laryngeal cancer, lung cancer, bladder cancer, prostate cancer, gastric cancer or malignant lymphoma.

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