Application of Dianemycin in in-vitro inhibition of TNBC cell activity
By developing the small molecule compound Dianemycin, it inhibits the proliferation and migration of TNBC cells and degrades the B7-H3 protein, the problems of limited treatment options and existing compounds are solved, and significant anti-tumor effects and good safety are achieved.
Patent Information
- Application Number
- CN202510283574.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-11
- Publication Date
- 2025-06-20
AI Technical Summary
The treatment options for triple-negative breast cancer (TNBC) are limited, traditional chemotherapy has limited efficacy and serious side effects. Existing small-molecule compounds face specificity, efficacy and safety issues in clinical applications.
A novel small molecule compound Dianemycin is developed to promote apoptosis by inhibiting the proliferation, migration and invasion of TNBC cells, and inhibit TNBC cell activity by degrading the B7-H3 protein.
Dianemycin significantly inhibits the proliferation and migration of TNBC cells, promotes apoptosis, has good safety and low toxicity, and provides a new TNBC targeted therapy strategy.
Smart Images
Figure CN120173882A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of pharmaceutical technology, and more specifically, it relates to the application of Dianemycin in inhibiting the activity of TNBC cells in vitro. Background Art
[0002] Triple-negative breast cancer (TNBC) is a highly aggressive subtype of breast cancer, characterized by negative expression of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2). Due to the lack of these biomarkers, the treatment options for TNBC are limited and it is insensitive to traditional hormone therapy and HER2-targeted therapy; currently, chemotherapy is the main treatment method for TNBC, but its efficacy is limited and it is accompanied by severe side effects.
[0003] Existing studies have shown that a variety of small molecule compounds and biological agents exhibit inhibitory effects on TNBC cells in in vitro experiments; for example, Tinengotinib (TT-00420) is a novel spectrum-selective small molecule kinase inhibitor with high activity against TNBC; however, these compounds still face many challenges in clinical applications, including issues such as drug specificity, efficacy, and safety.
[0004] Therefore, the development of new TNBC treatment drugs, especially compounds with a clear mechanism of action and good safety, is of great significance for improving the treatment effect and quality of life of TNBC patients; and Dianemycin, as a novel small molecule compound, its potential to inhibit the activity of TNBC cells in vitro has not been fully explored. Therefore, the present invention aims to provide the application of Dianemycin in inhibiting the activity of TNBC cells in vitro, in order to provide a new strategy for the treatment of TNBC. Summary of the Invention
[0005] The purpose of the present invention is to provide the application of Dianemycin in inhibiting the activity of TNBC cells in vivo and in vitro. The present invention illustrates that the compound Dianemycin can effectively inhibit the proliferation, migration, and invasion of TNBC cells and promote the apoptosis of TNBC cells by providing the application of a small molecule compound Dianemycin in inhibiting the activity of TNBC cells.
[0006] The above technical objective of the present invention is achieved through the following technical solutions:
[0007] The present invention provides the application of Dianemycin in inhibiting the activity of TNBC cells.
[0008] The present invention also provides the application of Dianemycin in the preparation of drugs for the treatment of TNBC.
[0009] The present invention is further configured such that: Dianemycin inhibits the proliferation, migration and invasion of TNBC cells, promotes the apoptosis of TNBC cells, and thus inhibits the cell activity of TNBC.
[0010] The present invention is further configured such that: Dianemycin inhibits the cell activity of TNBC by promoting the degradation of B7-H3.
[0011] The present invention also provides a method for treating TNBC, which includes administering an effective amount of Dianemycin to a patient.
[0012] In summary, the present invention has the following beneficial effects:
[0013] 1. The present invention verifies through in vitro experiments that Dianemycin can significantly inhibit the proliferation, migration and invasion of triple-negative breast cancer cells, and at the same time can promote the apoptosis of Dianemycin cells;
[0014] 2. In the in vivo experiments of the present invention, it is shown that Dianemycin does not show obvious toxicity. At the same time, by monitoring the body weight change, detecting indicators such as AKP / ALT / AST in the serum, and observing the HE staining sections of various organs, it is found that Dianemycin has good safety;
[0015] 3. The present invention finds the target of Dianemycin - B7-H3 through mass spectrometry and CETSA experiments, and through WB experiments, it is found that Dianemycin can promote the degradation of B7-H3, which can provide a new target for the targeted treatment of TNBC;
[0016] 4. The present invention clarifies the role of B7-H3 in tumors and its prognostic significance. It is known that B7-H3 is one of the co-stimulatory molecules of the B7 family, and its high expression is presented in various tumor tissues, including gastric cancer, lung cancer, prostate cancer, kidney cancer, breast cancer, etc. Research shows that the high expression of B7-H3 is closely related to tumor progression, metastasis and poor prognosis. For example, in non-small cell lung cancer, B7-H3 can promote the migration and invasion of tumor cells. In addition, B7-H3 also participates in tumor angiogenesis, further supporting tumor growth;
[0017] 5. The present invention also clarifies the ADC drugs targeting B7-H3 and their limitations. Currently, the targeted therapy for B7-H3 in clinical practice mainly focuses on antibody-drug conjugates (ADCs). ADC drugs achieve precise targeting of tumor cells by conjugating antibodies with cytotoxic drugs. However, ADC drugs have some limitations: Firstly, the antibody part of ADC drugs may have immunogenicity, leading to immune responses. Secondly, insufficient stability of the linker may cause premature release of cytotoxic drugs, increasing side effects. In addition, the production cost of ADC drugs is relatively high, and their efficacy is limited in certain tumors. In contrast, small molecule drugs have unique advantages. They generally have better tissue penetration, can reach tumor cells more effectively, and have lower production costs.
[0018] 6. The present invention also clarifies the research progress of the small molecule drug Dianemycin and finds that the small molecule drug Dianemycin can specifically degrade the B7-H3 protein, thereby inhibiting its pro-cancer effect in tumors. Using triple-negative breast cancer (TNBC) as a model, the present invention systematically studied the anti-tumor activity and its molecular mechanism of Dianemycin in vitro and in vivo. The research results show that Dianemycin can significantly inhibit the proliferation and migration of TNBC cells in vitro experiments and block its downstream signaling pathway by degrading the B7-H3 protein. In in vivo experiments, Dianemycin showed significant anti-tumor effects through a mouse xenograft model. This drug can inhibit tumor growth and no obvious toxicity was observed at a certain dose. This indicates that Dianemycin has good safety.
[0019] 7. The present invention also clarifies the application prospect of Dianemycin. Based on the research results of the present invention, it is clear that Dianemycin has broad application prospects in tumor treatment. Its mechanism of specifically degrading the B7-H3 protein enables it to act precisely on tumor cells, while avoiding the immunogenicity and non-specific toxicity of ADC drugs. In addition, the excellent performance of Dianemycin in the triple-negative breast cancer model provides new ideas for the treatment of other tumors with high expression of B7-H3. BRIEF DESCRIPTION OF THE DRAWINGS
[0020] Figure 1 It is a graph showing the experimental results of the inhibitory effect of Dianemycin on the activity of TNBC cells in vitro in Example 1 of the present invention;
[0021] Figure 2 It is a graph showing the experimental results of the verification of the in vivo function and safety of Dianemycin in Example 2 of the present invention;
[0022] Figure 3It is the experimental result graph of Dianemycin promoting the degradation of B7-H3 in Example 3 of the present invention. Detailed implementation manners
[0023] The following further elaborates on the present invention in conjunction with the attached Figures 1-3 drawings.
[0024] Example 1: Experiment on the in vitro inhibition of TNBC cell activity by Dianemycin
[0025] In this example, flow cytometry apoptosis detection was performed on cells treated with Dianemycin. Compared with the control group, the results were as Figure 1 shown, and Dianemycin significantly induced apoptosis in TNBC cells ( Figure 1 A); the increased expression of cleaved PARP and cleaved caspase-9 in TNBC cells treated with Dianemycin also confirmed the above results ( Figure 1 B); moreover, through the colony formation assay, it was found that Dianemycin could significantly reduce the proliferation ability of MDA-MB-231 and MDA-MB-468 cells at low concentrations ( Figure 1 C); in addition, Dianemycin could also significantly impair the migration and invasion abilities of MDA-MB-231 and MDA-MB-468 cells ( Figure 1 D).
[0026] Experimental conclusion: The small molecule Dianemycin can inhibit the proliferation, migration and invasion of TNBC cells, and at the same time promote the apoptosis of TNBC cells.
[0027] Example 2: In vivo function and safety verification experiment of Dianemycin
[0028] In this example, MDA-MB-468 cells were orthotopically injected into nude mice. When the tumors were palpable, the mice were randomly divided into 2 groups (n = 5 in each group), an untreated control group (DMSO) and a Dianemycin treatment group (2 mg / kg); Dianemycin or DMSO was intraperitoneally injected every other day for 20 consecutive days; during the experiment, the body weight and tumor size of the mice were monitored every two days. As Figure 2 shown, the tumors in the control group grew rapidly, but Dianemycin significantly inhibited tumor growth ( Figure 2 A and Figure 2 B); however, Dianemycin had no effect on the body weight ( Figure 2 C), alkaline phosphatase (AKP), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) of the mice ( Figure 2 D, Figure 2 E andFigure 2 F); At the same time, mouse heart, liver, spleen, lung, and kidney tissues were taken for H&E staining, and no obvious morphological or structural changes were observed ( Figure 2 G).
[0029] In summary, the experimental data of this example indicate that Dianemycin is a promising candidate drug with low toxicity and acceptable anti-cancer effects.
[0030] Experimental conclusion: Dianemycin can inhibit the growth of xenograft tumors. In this example, by monitoring body weight changes, detecting indicators such as AKP / ALT / AST in serum, and observing HE staining sections of various organs, it was found that Dianemycin has no obvious toxicity.
[0031] Example 3: Experiment on Dianemycin promoting the degradation of B7-H3
[0032] To find the target molecule of Dianemycin, a Darts experiment was conducted in this example. Cells were collected, lysed with NP40 (protease and phosphatase inhibitors had been added), and centrifuged to obtain the protein supernatant. The protein supernatant was divided into two groups equally, a drug-treated group and a non-drug-treated group; a certain concentration of Dianemycin or the same volume of DMSO was added simultaneously, and incubated at room temperature for 1 h; then each group was further divided into a protease K-added group and a non-added group, and protease K or enzyme-free ddH2O was added according to a certain ratio (mass ratio), and degraded at room temperature for 5 - 10 min; protein loading buffer Loading was added, and incubated at 95 °C for 10 min to terminate the degradation; the protein samples of the drug-treated group and the non-drug-treated group were stained with Coomassie Brilliant Blue to obtain differential bands ( Figure 3 A), and the differential bands were subjected to mass spectrometry sequencing. Finally, by screening for proteins that were not present in the non-drug-treated group but detected in the drug-treated group, potential targets were determined ( Figure 3 B). Then, through the CETSA experiment, it was found that at 44 °C, the B7-H3 protein in the Dianemycin-treated group was more stable than that in the control group, further proving that B7-H3 is the target molecule of Dianemycin ( Figure 3 C). According to the potential target molecules we screened, the cells were treated with different concentrations of Dianemycin, and the target molecule B7-H3 we screened was detected by WB experiment. It was found that Dianemycin could reduce the expression of B7-H3 and showed a certain concentration dependence ( Figure 3 D).
[0033] Experimental conclusion: Using mass spectrometry to find the potential target molecule of Dianemycin, B7-H3 that can be inhibited by Dianemycin was determined by WB experiment in this example.
[0034] This specific embodiment is only an interpretation of the present invention and does not limit the present invention. After reading this specification, those skilled in the art can make modifications to this embodiment without creative contributions as needed, but as long as it is within the scope of the claims of the present invention, it is protected by the patent law.
Claims
1. Application of Dianemycin in inhibiting TNBC cell activity.
2. Application of Dianemycin in the preparation of drugs for the treatment of TNBC.
3. The use of Dianemycin in inhibiting TNBC cell activity according to claim 1, characterized in that: The dianemycin inhibits the proliferation, migration and invasion of TNBC cells and promotes apoptosis of TNBC cells, thereby inhibiting the cell activity of TNBC.
4. The use of Dianemycin in inhibiting TNBC cell activity according to claim 1, characterized in that: The dianemycin inhibits the cell activity of TNBC by promoting the degradation of B7-H3.
5. A method for treating TNBC, characterized by: This includes administering to the patient an effective amount of Dianemycin.