Triazole compound and application thereof in breast cancer treatment

Abstract

The invention discloses a triazole compound and application thereof in breast cancer treatment, and relates to the technical field of medicinal chemistry, and the triazole compound is characterized in that the triazole compound inhibits the activity and proliferation of breast cancer cells in a concentration-dependent manner, reduces the migration ability, tumorigenicity and invasion ability of the breast cancer cells, and improves the breast cancer treatment effect. The apoptosis of breast cancer cells is induced; according to the triazole compound, the expression levels of apoptosis-related proteins Bcl-2, Bax and Caspase3 are regulated, and a breast cancer cell apoptosis signal channel is activated, so that the inhibition effect on breast cancer cells is achieved, and the breast cancer cells are promoted to be converted to the apoptosis direction. The activity and proliferation of breast cancer cells are inhibited through the newly synthesized triazole compound, the migration ability, tumorigenicity and invasion ability of the cells are reduced, the transformation of the breast cancer cells to the apoptosis direction is promoted, and a foundation can be laid for developing a more effective breast cancer treatment scheme.

Description

Technical Field

[0001] The present invention relates to the technical field of medicinal chemistry, and more specifically, it relates to a triazole compound and its application in the treatment of breast cancer. Background Art

[0002] As one of the most common malignant tumors in women globally, the incidence and mortality rates of breast cancer have been consistently high, seriously threatening the lives and health of women. Although current treatment methods, such as surgery, radiotherapy, chemotherapy, endocrine therapy, and targeted therapy, have improved the survival situation of patients to a certain extent, there are still many challenges, such as the emergence of drug resistance, treatment side effects, and recurrence and metastasis. Therefore, finding new effective therapeutic drugs and strategies has become an important direction in breast cancer research.

[0003] In recent years, the research and development of new anti-cancer drugs have become a research hotspot. Due to their unique chemical structures and biological activities, triazole compounds have shown potential application values in the anti-tumor field. Research has shown that such compounds can act on tumor cells through multiple mechanisms. They can interfere with the cell cycle progression of tumor cells, leading to cell cycle arrest, thereby inhibiting the proliferation of tumor cells. In addition, they can also regulate signal pathways within tumor cells, such as the PI3K / Akt / mTOR pathway, affecting the survival and 5-fluorouracil metabolism of tumor cells. At the same time, some triazole compounds can induce apoptosis in tumor cells, promoting tumor cell death by activating endogenous and exogenous apoptosis pathways.

[0004] Apoptosis is a physiological cell death program, which is crucial for maintaining cell homeostasis and tissue balance. Abnormal regulation of apoptosis is closely related to the occurrence and development of tumors. Inducing apoptosis in tumor cells has become one of the important strategies in the research and development of anti-cancer drugs.

[0005] Therefore, the present invention aims to provide a triazole compound and its application in the treatment of breast cancer. By inhibiting the viability and proliferation of breast cancer cells with newly synthesized triazole compounds, reducing the cell migration ability, tumorigenicity, and invasion ability, and promoting the transformation of breast cancer cells towards apoptosis. Summary of the Invention

[0006] The object of the present invention is to provide a triazole compound and its application in the treatment of breast cancer. By inhibiting the viability and proliferation of breast cancer cells with newly synthesized triazole compounds, reducing the cell migration ability, tumorigenicity, and invasion ability, and promoting the transformation of breast cancer cells towards apoptosis, it can lay a foundation for the development of more effective breast cancer treatment regimens.

[0007] The present invention provides an application of a triazole compound in the treatment of breast cancer.

[0008] The present invention also provides an application of a triazole compound in the preparation of a drug for treating breast cancer.

[0009] The present invention is further configured such that: the triazole compound inhibits the viability and proliferation of breast cancer cells in a concentration-dependent manner, reduces the migration ability, tumorigenicity and invasion ability of breast cancer cells, and induces apoptosis of breast cancer cells.

[0010] The present invention is further configured such that: the triazole compound exerts an inhibitory effect on breast cancer cells and promotes the transformation of breast cancer cells towards apoptosis by regulating the expression levels of apoptosis-related proteins Bcl-2, Bax and Caspase3 and activating the apoptosis signaling pathway of breast cancer cells.

[0011] The present invention is further configured as:

[0012] The structural formula of the triazole compound is:

[0013]

[0014] The present invention also provides a drug composition, comprising the triazole compound and a pharmaceutical carrier.

[0015] In summary, the present invention has the following beneficial effects:

[0016] 1. The triazole compound yu10118 in the present invention can significantly inhibit the viability and proliferation of breast cancer cells in a concentration-dependent manner, and at the same time can reduce the migration ability, tumorigenicity and invasion ability of breast cancer cells, and induce apoptosis of breast cancer cells, thereby restricting the spread and metastasis of cancer cells;

[0017] 2. The triazole compound yu10118 in the present invention exerts an inhibitory effect on breast cancer cells by regulating the expression levels of apoptosis-related proteins Bcl-2, Bax and Caspase3, activating the apoptosis signaling pathway of breast cancer cells, and promoting the transformation of breast cancer cells towards apoptosis, so that the triazole compound yu10118 can provide new ideas for the treatment of breast cancer and serve as a potential drug target, which helps to develop more effective treatment regimens;

[0018] 3. The triazole compound yu10118 in the present invention shows potential application value in the field of anti-tumor. The drug prepared from the triazole compound yu10118 can play an important role in the treatment of breast cancer and has potential advantages in inhibiting tumor cell proliferation, reducing metastasis ability, inducing cell apoptosis and improving the sensitivity of chemotherapy drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1It is the result diagram of the effect of yu10118 on the viability of breast cancer cells in Example 1 of the present invention;

[0020] Figure 2 It is the result diagram of the effect of yu10118 on the invasion ability of breast cancer cells in Example 2 of the present invention;

[0021] Figure 3 It is the result diagram of the effect of yu10118 on the protein expression of MDA-MB-231 and MCF-7 in Example 3 of the present invention. Detailed implementation manners

[0022] The following further describes the present invention in detail with reference to the appended Figures 1 - 3 drawings.

[0023] The cell lines and main reagents used in the embodiments of the present invention are as follows:

[0024] Human breast cancer MBA-MD-231 and MCF-7 cell lines were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China); DMEM / F12 culture medium and DMEM culture medium were purchased from Gibco Company, USA; 5-fluorouracil (5-Fu), trypsin cell digestive solution, DMSO solvent, and BCA protein quantification kit were purchased from Beyotime Institute of Biotechnology, China; penicillin-streptomycin solution and cell culture grade fetal bovine serum were purchased from Sangon Biotech Co., Ltd., China; Hoechst 33342 was purchased from MERCK Company, Germany.

[0025] The cell culture process in this embodiment is as follows: Human breast cancer cell lines MBA-MD-231 and MCF-7 were selected and cultured in DMEM medium and DMEM / F12 medium containing 10% fetal bovine serum and 1% penicillin-streptomycin, respectively, and the cells were placed in an incubator at 37°C and 5% CO2. The medium was changed to fresh medium every 1-2 days. When the cell confluence reached about 80%-90%, subculture or cryopreservation was carried out.

[0026] Regarding cell grouping and drug administration, yu10118 powder was dissolved in dimethyl sulfoxide (DMSO), and a stock solution of 200 μmol / L was prepared through a 0.45 μm filter membrane and stored in a -20°C refrigerator. When used, the stock solution was diluted with DMSO to the corresponding concentration for the experiment. 5-fluorouracil (5-FU) was stored in the same way. In this embodiment, the cells were divided into a blank control group, an experimental group, and a positive control group. The cells in the blank control group were only cultured in normal culture medium without adding any drugs; the cells in the experimental group were treated with different concentrations of yu10118 solution; 5-Fu is a chemotherapy drug widely used in the treatment of breast cancer, and it was selected as the positive control.

[0027] In this example, statistical analysis was performed on the experimental results. One-way ANOVA was carried out using SPSS software. The data were presented as mean ± SD. "*" indicates that the P value is less than 0.05 and is considered statistically significant.

[0028] Example 1: Cell Proliferation Assay

[0029] In this example, cells in the logarithmic growth phase were collected, and the density of the cell suspension was adjusted to 2×10 4 cells / mL and dispensed into 96-well plates at 100 μL per well. Then, the plates were placed in a cell culture incubator at 37 °C and 5% CO2 to allow the cells to adhere well. After the cells adhered, the old culture medium was discarded, and the drug-containing culture medium with gradient dilution was added. After 24 h and 48 h respectively, the drug-containing culture medium was discarded, and fresh culture medium containing 10% CCK-8 was added, and incubation was continued for 1 h. The absorbance at 450 nm was measured using a microplate reader, and the cell viability was calculated. Cell viability = (absorbance of cells in the blank group - absorbance of cells in the experimental group) / absorbance of cells in the blank group × 100%.

[0030] The results of the effect of yu10118 on the viability of breast cancer cells in this example are as follows:

[0031] According to the results of the CCK-8 method, in the MDA-MB-231 cell line, after yu10118 treatment for 24 h and 48 h respectively, as the concentration of yu10118 gradually increased, the cell viability gradually decreased. As Figure 1 shown, (a) shows the cell viability of MDA-MB-231 cells after drug treatment for 24 h and 48 h; (b) shows the cell viability of MCF-7 cells after treatment for 24 h and 48 h; (c) shows the half-maximal inhibitory concentration (IC50) of MDA-MB-231 cells after drug treatment for 48 hours; (d) shows the half-maximal inhibitory concentration (IC50) of MCF-7 cells after drug treatment for 48 hours. The bar graphs represent the mean ± standard deviation (n = 3). 5-Fluorouracil (5-Fu) was used as a positive control, and "*" indicates significant difference (P < 0.05).

[0032] In the MCF-7 cell line, when yu10118 acted continuously for 24 h, yu10118 at 25 μmol / L had no obvious effect on cell viability (P > 0.05), but after treatment for 48 h, cell viability was significantly inhibited ( Figure 1 b). As a positive control, 5-FU also showed a strong inhibitory effect on the proliferation viability of the two cell lines ( Figure 1 a and Figure 1b), The IC50 values of yu10118 and 5-Fu against MDA-MB-231 cells after 48 h of treatment were 58.58 ± 2.87 μmol / L and 11.68 ± 4.92 μmol / L, respectively ( Figure 1 c), The IC50 values against MCF-7 cells after 48 h of treatment were 49.03 ± 5.25 μmol / L and 34.97 ± 5.07 μmol / L, respectively ( Figure 1 d).

[0033] Example 2: Cell invasion experiment

[0034] The upper and lower chambers of the invasion chamber were hydrated with basal culture medium for 1 h respectively. The hydrated culture medium in the small chamber was discarded. 300 μl of cell suspension at a density of 1.5×10 5 cells / ml after 48 h of drug treatment was added to the upper chamber. At the same time, 600 μl of complete culture medium containing fetal bovine serum was added to the lower chamber; after overnight incubation, the upper chamber was taken out alone, replaced with basal culture medium containing different concentrations of drugs, and then immediately placed back into the original well and cultured for another 48 h; the excess cells in the upper chamber were wiped off with a cotton swab and washed twice with PBS; fixed with 4% paraformaldehyde for 15 min and stained with 0.1% crystal violet for 20 min; the small chamber was inverted and observed and photographed under a microscope.

[0035] The results of the effect of yu10118 on the invasion ability of breast cancer cells in this example are as follows:

[0036] As Figure 2 shown, (a) shows the invasion of MDA-MB-231 cells after 48 h of treatment with yu10118, and the scale bar is 200 μm; (b) shows the invasion numbers of MDA-MB-231 cells at different drug concentrations; (c) shows the invasion of MCF-7 cells after 48 h of treatment with yu10118, and the scale bar is 200 μm; (d) shows the invasion numbers of MCF-7 cells; the results show that after 48 h of drug treatment, yu10118 can significantly reduce the invasion ability of MDA-MB-231 cells, and the number of cells passing through the basement membrane is significantly reduced ( Figure 2 a, Figure 2 b). At the same time, for MCF-7 cells, yu10118 also showed a strong inhibitory effect on invasion. Compared with the control group, the degree of cell invasion in the treatment group was significantly weakened ( Figure 2 c, Figure 2 d). Further observation found that with the increase of the concentration of yu10118, the invasion ability of both types of cells showed a gradually decreasing trend, indicating that yu10118 has an effective inhibitory effect on the invasion of MDA-MB-231 and MCF-7 cells, providing an important basis for further studying its anti-tumor mechanism.

[0037] Example 3: Western blot experiment

[0038] After treating MDA-MB-231 and MCF-7 cells with the drug-containing culture medium for 48 h, the cells were collected, an appropriate amount of protein lysate was added to lyse the cells on ice, and then centrifuged at 12,000 rpm for 10 min at 4 °C. The supernatant was collected, which was the total protein. The protein concentration was measured using a BCA protein quantification kit, and the protein concentrations of each sample were adjusted to be consistent. The protein samples were mixed with the loading buffer, heat-denatured, and then subjected to SDS-PAGE gel electrophoresis to separate the proteins according to their molecular weights. The proteins in the gel were transferred to a PVDF membrane by wet transfer. Subsequently, the non-specific binding sites on the membrane were blocked with 5% skim milk, and then the primary antibody incubation step was carried out: mouse anti-GAPDH (1:2500), mouse anti-BAX (1:2000), rabbit anti-Bcl-2 (1:2000), rabbit anti-Caspase-3 (1:2000), incubated at 4 °C for 12 h. Subsequently, the membrane was washed to remove the unbound primary antibody, and then HRP-labeled secondary antibody was added and incubated at room temperature for 1 h. The membrane was washed again to remove the unbound secondary antibody. BeyoECL Moon chemiluminescent substrate was evenly dropped on the membrane and placed in an imager for exposure and color development to obtain an image. Using GAPDH as the protein internal reference, the gray value of the protein band was analyzed using Image J software to obtain the protein expression level.

[0039] The results of the effect of yu10118 on the protein expression of MDA-MB-231 and MCF-7 in this example are as follows:

[0040] As Figure 3 shown, (a) shows the detection of the activities of Caspase 3 (cysteine aspartic protease 3), Bcl-2 (B-cell lymphoma-2 protein), and Bax (Bcl-2-associated X protein) after treating MDA-MB-231 cells with yu10118; (b) shows the quantitative determination of each protein level using Image-J software and normalization with GAPDH (glyceraldehyde-3-phosphate dehydrogenase) as a reference; (c) shows the expression status of related proteins in MCF-7 cells; (d) shows the quantitative determination of the protein level using Image-J software; the results of the Western Blot experiment show that yu10118 has a significant effect on the expression of Caspase3, bcl-2, and Bax proteins in MDA-MB-231 and MCF-7 breast cancer cells ( Figure 3 a, Figure 3c), Caspase3 is a key execution protein in the process of apoptosis. An increase in the total protein expression in the two cell lines indicates that the apoptosis program may be initiated. The Bax protein can promote apoptosis, and the increase in its expression in the experiment further confirms that yu10118 can induce breast cancer cells to apoptosis. While the Bcl-2 protein usually plays an anti-apoptotic role, and its expression decreased in this experiment, indicating that yu10118 weakens the anti-apoptotic ability of breast cancer cells. These changes in protein expression together suggest that yu10118 may affect the fate of breast cancer cells by regulating the balance of apoptosis-related proteins.

[0041] In summary, the present invention shows that yu10118 has a significant inhibitory effect on breast cancer cells. Its mechanism is related to the regulation of the expression of apoptosis-related proteins, which can provide new hope and research directions for the treatment of breast cancer.

[0042] This specific embodiment is only an interpretation of the present invention, and it is not a limitation of the present invention. Those skilled in the art can make modifications without creative contributions to this embodiment according to needs after reading this specification, 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 a triazole compound in the treatment of breast cancer.

2. Use of a triazole compound in the preparation of a drug for treating breast cancer.

3. The use of a triazole compound in the treatment of breast cancer according to claim 1, characterized in that: The triazole compound inhibits the activity and proliferation of breast cancer cells in a concentration-dependent manner, reduces the migration ability, tumorigenicity and invasion ability of breast cancer cells, and induces apoptosis of breast cancer cells.

4. The use of a triazole compound in the treatment of breast cancer according to claim 3, characterized in that: The triazole compound activates the apoptosis signaling pathway of breast cancer cells by regulating the expression levels of apoptosis-related proteins Bcl-2, Bax and Caspase3, thereby inhibiting breast cancer cells and promoting the transformation of breast cancer cells to apoptosis.

5. A triazole compound, characterized in that: The structural formula of the triazole compound is:

6. A pharmaceutical composition characterized by: The invention comprises the triazole compound as claimed in claim 5 and a pharmaceutically acceptable carrier.

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