Application of furazidin in antitumor drugs
By targeting and inhibiting the phosphorylation process of STAT3 protein, cyproterone acetate has solved the problem of insufficient performance of existing anti-tumor drugs in the treatment of lung cancer, cervical cancer and ovarian cancer, achieving high efficiency in inhibiting these cancer cells and low toxicity to normal cells, showing excellent anti-tumor performance.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- INST OF MATERIA MEDICA CHINESE ACAD OF MEDICAL SCI
- Filing Date
- 2024-11-26
- Publication Date
- 2026-05-26
AI Technical Summary
Existing broad-spectrum antitumor drugs, such as fluorouracil, have insufficient antitumor performance in the treatment of cancers such as lung cancer, cervical cancer, and ovarian cancer, and have poor safety and selectivity for normal cells.
Using cyproterenol as an anti-tumor drug, it inhibits the phosphorylation of STAT3 protein by targeting and preventing p-STAT3 dimer from entering the cell nucleus, thereby inhibiting the expression of BCL-2 protein. It mainly acts on the G2/M phase of tumor cells to inhibit the proliferation of tumor cells.
Glycoside showed superior antitumor activity compared to fluorouracil in various cancer cell lines, especially against lung cancer, cervical cancer and ovarian cancer cells. It also showed no significant toxicity to normal cells at low concentrations, exhibiting concentration- and time-dependent effects and significantly inhibiting the proliferation of tumor cells.
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Abstract
Description
Technical Field
[0001] This invention belongs to the field of pharmaceutical technology, specifically the field of biomedical technology concerning the inhibitory effect of cytosine on tumor cells, and more specifically, the application development and mechanism of action of cytosine on the inhibitory effects of lung cancer, cervical cancer and ovarian cancer. Background Technology
[0002] Cancer is a major public health and economic issue in the 21st century, accounting for nearly one-fifth of all non-communicable disease deaths worldwide. [1-3] Among these, treatment options for lung cancer—the second most common cancer in the world—and cervical cancer and ovarian cancer—the most prevalent gynecological malignancies among women in my country—are currently hot topics in clinical research. [4-7] In this study, we used fluorouracil, a broad-spectrum antitumor drug used in clinical applications, as a control. Based on the excellent half-maximal inhibitory concentration (IC50) results of cyhalothrin against lung cancer cells A549, cervical cancer cells HeLa, and ovarian cancer cells A2780, we directly verified the anticancer target of cyhalothrin through experiments, elucidated the anticancer mechanism of cyhalothrin, and provided theoretical and experimental basis for its entry into preclinical and clinical trials. Summary of the Invention
[0003] The purpose of this invention is to develop the application and mechanism of action of sclerotinib in tumor inhibition. This invention successfully discovered that sclerotinib possesses broad-spectrum antitumor activity, including against pancreatic cancer cells, gastric cancer cells, breast cancer cells, ovarian cancer cells, colon cancer cells, cervical cancer cells, liver cancer cells, and lung cancer cells, with particularly superior effects in inhibiting the growth of lung cancer cells, cervical cancer cells, and ovarian cancer cells. Compared to the currently available broad-spectrum antitumor drug fluorouracil, sclerotinib exhibits superior antitumor properties. Further research revealed that sclerotinib acts on the novel protein STAT3 in cancer treatment, inhibiting STAT3 phosphorylation and preventing phosphorylated protein dimers from entering the cell nucleus to initiate transcription, thereby inhibiting tumor cell proliferation. In addition, sclerotinib also inhibits the downstream anti-apoptotic protein BCL-2 of STAT3. Further research showed that the inhibitory effect of sclerotinib on cancer cells keeps them primarily in the G2 / M phase of cell proliferation, synergistically inhibiting tumor cell growth and proliferation.
[0004] To achieve the above objectives, in a first aspect, the present invention provides screening results of cytosine for 8 cancer cell lines and 25 tumor cell types.
[0005] Secondly, this invention provides a biosafety evaluation of cytosine in normal human cells.
[0006] Thirdly, this invention provides that the inhibitory effect of styrosine on lung cancer cells A549, cervical cancer cells HeLa, and ovarian cancer cells A2780 is concentration-dependent.
[0007] Fourthly, this invention provides STAT3, a target for the anticancer activity of styraxetin verified by Western blotting experiments, which inhibits the phosphorylation process of STAT3 and exhibits both time- and concentration-dependent effects.
[0008] Fifthly, the present invention provides that the inhibitory effect of cytosine on cancer cells, as determined by flow cytometry, is mainly in the G2 / M phase of cell proliferation.
[0009] Beneficial technical effects:
[0010] Firstly, a total of 8 cancer cell lines and 25 types of tumor cells were screened. Among all the screened tumor cells, cyproterone acetate had a stronger effect on tumor cells than fluorouracil, with the strongest inhibitory effect in tumor cells A549, HeLa, and A2780.
[0011] Secondly, when the concentration of cyproterone was 0.08 μM and the treatment time was 24 h, it did not have significant toxicity to four types of normal human cells.
[0012] Thirdly, when the concentration of cyhalothrin was 0.08 μM and the treatment time was 24 h, it showed inhibitory effects on all three types of tumor cells, with significant differences in activity among the three types. Furthermore, the inhibitory effect of cyhalothrin on lung cancer cells A549, cervical cancer cells HeLa, and ovarian cancer cells A2780 was concentration-dependent.
[0013] Fourthly, in A549, HeLa, and A2780 cells, cyproheptadine at a concentration of 0.08 μM significantly inhibited the expression of p-STAT3, indicating that cyproheptadine inhibited STAT3 phosphorylation, prevented p-STAT3 dimers from entering the cell nucleus, and thus inhibited the expression of downstream BCL-2 protein, thereby inhibiting the proliferation of tumor cells.
[0014] Fifthly, in A549, Hela, and A2780 cells, cytosine inhibited cell proliferation during the G2 / M phase, and in A549 and Hela cells, this inhibition was concentration-dependent. Attached Figure Description
[0015] Figure 1 This indicates the screening results of the inhibitory effect of cytosine on eight cancer cell lines.
[0016] Figure 2 This indicates that the standard CCK-8 method was used to determine the toxicity of cytosine to normal human cells.
[0017] Figure 3 The study indicated that the inhibitory effects of different concentrations of salicylic acid on lung cancer cells, cervical cancer cells, and ovarian cancer cells were determined using the standard CCK-8 assay.
[0018] Figure 4 This indicates that Western blotting experiments were used to verify the anticancer target of styrosine.
[0019] Figure 5 This indicates that flow cytometry was used to determine the inhibitory effect of cytosine on cancer cells. Detailed Implementation
[0020] The following provides a detailed description of specific embodiments of the present invention. It should be understood that the specific embodiments described herein are for illustrative and explanatory purposes only and are not intended to limit the scope of the invention.
[0021] Example 1: Screening for the inhibitory effect of cytosine on cancer cell lines. Antitumor activity was screened on 25 tumor cell lines from 8 cancer cell lines, including pancreatic cancer (Panc-1, BXPC-3, SW1990), gastric cancer (HGC-27, MKN45, AGS), breast cancer (MCF-7, MDA-MB-231, MDA-MB-468), ovarian cancer (A2780, OVCAR-3, SK-OV-3), colon cancer (HCT-116, DLD1, HT-29, SW480), cervical cancer (HeLa, C-c33 A, SiHa), liver cancer (HUH-7, HepG2, Hep 3B), and lung cancer (A549, H1299, NCI-H460).
[0022] I. Experimental Instruments and Reagents
[0023] Instruments: CO2 incubator: Thermo; centrifuge: Sigma, Germany; biosafety cabinet: Haier; EnVision: PerkinElmer; 96-well cell culture plate: SORFA.
[0024] The test drugs were: fluorouracil (FFN) purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd.; and cyproterone acetate purchased from Chengdu Yirui Biotechnology Co., Ltd.
[0025] II. Preparation of Cell Growth Culture Medium
[0026] Prepare various cell growth media under aseptic conditions, store the media in a 4°C refrigerator for later use, and preheat to 37°C before use.
[0027] III. Cell Passaging and Seeding
[0028] When the cells reached 80-90% confluence in the culture dish, they were digested with trypsin and EDTA solution, centrifuged, and then resuspended in fresh culture medium. The cells were then passaged at appropriate ratios. Specifically, HCT-116, HT-29, and HeLa were passaged at a 1:5 ratio; SW480, DLD1, A2780, and SK-OV-3 at a 1:4 ratio; H1299, NCI-H460, AGS, HGC-27, OVCAR-3, BXPC-3, MKN45, A549, MCF-7, HepG2, Hep 3B, SiHa, C-33A, C-33A, MDA-MB-231, and HUH-7 at a 1:3 ratio; and MDA-MB-468 and SW1990 at a 1:2 ratio.
[0029] One day before the assay, HCT-116, HT-29, DLD-1, A549, HeLa, and OVCAR-3 cells were seeded at 2000 cells / well in 96-well cell culture plates. SW480, MDA-MB-231, and Hep cells were seeded at 2000 cells / well. 3B, MKN45, HGC-27, SK-OV-3, A2780, and Panc-1 were seeded at 3000 cells / well in 96-well cell culture plates; MCF-7, H1299, NCI-H460, AGS, SiHa, and C-33A were seeded at 4000 cells / well in 96-well cell culture plates; HepG2, HUH-7, and BXPC-3 were seeded at 5000 cells / well in 96-well cell culture plates; and MDA-MB-468 and SW-1990 were seeded at 6000 cells / well in 96-well cell culture plates. 80 μL of cell suspension was seeded in each well. The cell culture plates were incubated at 37°C in a 5% CO2 cell culture incubator for 24 hours.
[0030] IV. Drug Treatment
[0031] The highest concentration of compound FFN was prepared to be 32.4 mM, and the highest concentration of compound PCG was prepared to be 0.162 mM. These were gradually diluted with culture medium to achieve the appropriate concentrations. After incubating the cells, the culture medium was removed, and each well was filled with the prepared culture medium containing different concentrations of the test compounds. Cells without the drug served as the control group, and the treated groups served as the treatment group. The cells were incubated at 37°C in a 5% CO2 cell culture incubator for 72 hours before the samples were collected.
[0032] After incubation, add 10 μL of CCK8 assay reagent to each well of the cell plate and incubate at 37°C for 0.5 h to 4 h. After incubation, measure the absorbance at 450 nm using a multi-functional microplate reader.
[0033] V. Data Processing
[0034] The logarithm of drug concentration was used as the X-axis and cell inhibition rate as the Y-axis for fitting, and the IC50 was calculated.50 The values are displayed on the chart.
[0035] VI. Experimental Results
[0036]
[0037] Table 1
[0038] Eight cancer cell lines and 25 types of tumor cells were screened. Among all the screened tumor cells, cytosine had a stronger effect on tumor cells than fluorouracil. The inhibitory effect was strongest in tumor cells A549, HeLa, and A2780. The specific results are shown in Table 1.
[0039] Example 2: The toxicity of cytosine to normal human cells was determined using the standard CCK-8 method.
[0040] I. Experimental Instruments and Reagents
[0041] Instruments: CO2 incubator: Thermo; centrifuge: Sigma, Germany; biosafety cabinet: Haier; EnVision: PerkinElmer; 96-well cell culture plate: SORFA.
[0042] Trial drug: Glycoside was purchased from Chengdu Yirui Biotechnology Co., Ltd.
[0043] II. Preparation of Cell Growth Culture Medium
[0044] Prepare various cell growth media under aseptic conditions, store the media in a 4°C refrigerator for later use, and preheat to 37°C before use.
[0045] III. Cell Passaging and Seeding
[0046] When the cells reach 80-90% confluence in the culture dish, digest them with trypsin and EDTA solution, centrifuge, and then resuspend the cells in fresh culture medium. Passage the cells at appropriate ratios. IOSE-80 and BEAS-2B are passaged at a 1:4 ratio, and HEK293 and THLE-2 are passaged at a 1:3 ratio.
[0047] One day prior to the assay, BEAS-2B cells were seeded at 10,000 cells / well, 3,000 cells / well, and 1,000 cells / well in 96-well cell culture plates, with 80 μL of cell suspension in each well. The plates were incubated at 37°C in a 5% CO2 incubator for 24 hours. IOSE-80 cells were seeded at 6,000 cells / well, 3,000 cells / well, and 1,000 cells / well in 96-well cell culture plates, with 80 μL of cell suspension in each well. The plates were incubated at 37°C in a 5% CO2 incubator for 24 hours. HEK293 cells were seeded at 8,000 cells / well, 2,000 cells / well, and 1,000 cells / well in 96-well cell culture plates, with 80 μL of cell suspension in each well. The plates were incubated at 37°C in a 5% CO2 incubator for 24 hours. THLE-2 cells were seeded in 96-well cell culture plates at concentrations of 15,000 cells / well, 5,000 cells / well, and 2,000 cells / well, with 80 μL of cell suspension inoculated into each well. The cell culture plates were incubated at 37°C in a 5% CO2 cell culture incubator for 24 hours.
[0048] IV. Drug Treatment
[0049] The highest concentration of compound PCG was prepared at 0.162 mM. It was gradually diluted with culture medium to achieve the appropriate concentration. After incubating the cells, the culture medium was removed, and different concentrations of the prepared culture medium containing the test compound were added to each well. Cells without the drug served as the control group, and the treated group served as the treatment group. Cells were incubated at 37°C in a 5% CO2 cell culture incubator for 24, 48, and 72 hours before being harvested.
[0050] After incubation, add 10 μL of CCK8 assay reagent to each well of the cell plate and incubate at 37°C for 2-3 hours. After incubation, measure the absorbance at 450 nm using a multi-functional microplate reader.
[0051] V. Data Processing
[0052] Substitute the measured absorbance into the following formula to calculate the cell viability.
[0053] Cell viability % = [(As-Ab) / (Ac-Ab)] × 100%
[0054] As: Absorbance of the experimental group (including cells, culture medium, CCK-8 solution and drug solution);
[0055] Ac: Absorbance of the control group (containing cells, culture medium, and CCK-8 solution, but excluding drugs);
[0056] Ab: Absorbance of blank wells (containing culture medium and CCK-8 solution, but excluding cells and drugs).
[0057] Use the R language to perform statistics and plot the graphs.
[0058] VI. Experimental Results
[0059] When the concentration of cyproterone was 0.08 μM and the treatment time was 24 h, it did not have significant toxicity to four types of normal human cells.
[0060] Example 3: The inhibitory effects of different concentrations of salicylic acid on lung cancer cells, cervical cancer cells and ovarian cancer cells were determined using the standard CCK-8 method.
[0061] I. Experimental Instruments and Reagents
[0062] Instruments: CO2 incubator: Thermo; centrifuge: Sigma, Germany; biosafety cabinet: Haier; EnVision: PerkinElmer; 96-well cell culture plate: SORFA.
[0063] Trial drug: Glycoside was purchased from Chengdu Yirui Biotechnology Co., Ltd.
[0064] II. Preparation of Cell Growth Culture Medium
[0065] Prepare various cell growth media under aseptic conditions, store the media in a 4°C refrigerator for later use, and preheat to 37°C before use.
[0066] III. Cell Passaging and Seeding
[0067] When the cells reach 80-90% confluence in the culture dish, digest them with trypsin and EDTA solution, centrifuge, and then resuspend the cells in fresh culture medium. Passage the cells at appropriate ratios: HeLa at a 1:5 ratio, A2780 at a 1:4 ratio, and A549 at a 1:3 ratio.
[0068] One day before the assay, A549 and HeLa cells were seeded at 2000 cells / well in 96-well cell culture plates, and A2780 cells were seeded at 3000 cells / well in 96-well cell culture plates. 80 μL of cell suspension was seeded in each well. The cell culture plates were incubated at 37°C in a 5% CO2 cell culture incubator for 24 hours. IV. Drug Treatment
[0069] The highest concentration of compound PCG was prepared to be 1 mM. It was gradually diluted with culture medium to achieve the appropriate concentration. After incubating the cells, the culture medium was removed, and each well was filled with prepared culture medium containing different concentrations of the test compound. Cells without the drug served as the control group, and the treated group served as the treatment group. The cells were incubated at 37°C in a 5% CO2 cell culture incubator for 24 hours before being collected.
[0070] After incubation, 10 μL of CCK8 assay reagent was added to each well of the cell plate, and the plate was incubated at 37°C for 2 hours. After incubation, the absorbance was measured at 450 nm using a multi-functional microplate reader.
[0071] V. Data Processing
[0072] Substitute the measured absorbance into the following formula to calculate the cell viability.
[0073] Cell viability % = [(As-Ab) / (Ac-Ab)] × 100%
[0074] As: Absorbance of the experimental group (including cells, culture medium, CCK-8 solution and drug solution);
[0075] Ac: Absorbance of the control group (containing cells, culture medium, and CCK-8 solution, but excluding drugs);
[0076] Ab: Absorbance of blank wells (containing culture medium and CCK-8 solution, but excluding cells and drugs).
[0077] Use the R language to perform statistics and plot the graphs.
[0078] VI. Experimental Results
[0079] At a concentration of 0.08 μM and an action time of 24 h, cytosine showed inhibitory effects on all three types of tumor cells, with significant differences observed in each cell type. Furthermore, the inhibitory effect of cytosine on lung cancer cells A549, cervical cancer cells HeLa, and ovarian cancer cells A2780 was concentration-dependent.
[0080] Experiment Example 4: Western Blot experiment to verify the anticancer target of styrax glycosides—at the same concentration but different times.
[0081] I. Experimental Instruments and Reagents
[0082] Instruments: Constant temperature incubator: Shanghai Yiheng; Centrifuge: Sigma, Germany; Biosafety cabinet: Haier; Protein vertical electrophoresis apparatus: Biorad, USA; Trans-Blot transfer tank: Biorad, USA; 96-well cell culture plate: SORFA.
[0083] Trial drug: Glycoside was purchased from Chengdu Yirui Biotechnology Co., Ltd.
[0084] II. Preparation of Cell Growth Culture Medium
[0085] Prepare various cell growth media under aseptic conditions, store the media in a 4°C refrigerator for later use, and preheat to 37°C before use.
[0086] III. Cell Passaging and Seeding
[0087] When the cells reach 80-90% confluence in the culture dish, digest them with trypsin and EDTA solution, centrifuge, and then resuspend the cells in fresh culture medium. Passage the cells at appropriate ratios: HeLa at a 1:5 ratio, A2780 at a 1:4 ratio, and A549 at a 1:3 ratio.
[0088] One day before the assay, A549 and HeLa cells were seeded at 2000 cells / well in 96-well cell culture plates, and A2780 cells were seeded at 3000 cells / well in 96-well cell culture plates. 80 μL of cell suspension was seeded in each well. The cell culture plates were placed in a cell culture incubator at 37°C and 5% CO2 and incubated for 24 hours.
[0089] IV. Drug Treatment
[0090] The highest concentration of compound PCG was prepared to be 1 mM. It was gradually diluted with culture medium to achieve the appropriate concentration. After incubating the cells, the culture medium was removed, and each well was filled with prepared culture medium containing different concentrations of the test compound. Cells without the drug served as the control group, and the treated group served as the treatment group. Cells were incubated at 37°C in a 5% CO2 cell culture incubator for 4–36 h before collection. Cells in the treated group were incubated at 37°C in a 5% CO2 cell culture incubator for 12–36 h before collection (n=3).
[0091] Western blotting was used to detect the expression of STAT3, p-STAT3, BCL-2, and β-Actin proteins in cell samples.
[0092] V. Data Processing
[0093] Substitute the measured protein expression level into the following formula to calculate the relative protein expression level.
[0094] Relative protein expression level = As / Ab
[0095] As: Protein expression level in the control group / experimental group;
[0096] Ab: β-Actin protein expression level.
[0097] Use the R language to perform statistics and plot the graphs.
[0098] VI. Experimental Results
[0099] p-STAT3 protein levels began to rise at 12 hours in the control group, therefore subsequent observations were conducted after 12 hours. In A549, HeLa, and A2780 cells, cyhalothrin at a concentration of 0.08 μM significantly inhibited p-STAT3 expression, indicating that cyhalothrin inhibited STAT3 phosphorylation, prevented p-STAT3 dimers from entering the nucleus, and thus inhibited the expression of downstream BCL-2 protein, thereby suppressing tumor cell proliferation.
[0100] Experiment Example 5: Western Blot experiment to verify the anticancer target of styrax glycosides at different concentrations at the same time.
[0101] I. Experimental Instruments and Reagents
[0102] Instruments: Constant temperature incubator: Shanghai Yiheng; Centrifuge: Sigma, Germany; Biosafety cabinet: Haier; Protein vertical electrophoresis apparatus: Biorad, USA; Trans-Blot transfer tank: Biorad, USA; 96-well cell culture plate: SORFA.
[0103] Trial drug: Glycoside was purchased from Chengdu Yirui Biotechnology Co., Ltd.
[0104] II. Preparation of Cell Growth Culture Medium
[0105] Prepare various cell growth media under aseptic conditions, store the media in a 4°C refrigerator for later use, and preheat to 37°C before use.
[0106] III. Cell Passaging and Seeding
[0107] When the cells reach 80-90% confluence in the culture dish, digest them with trypsin and EDTA solution, centrifuge, and then resuspend the cells in fresh culture medium. Passage the cells at appropriate ratios: HeLa at a 1:5 ratio, A2780 at a 1:4 ratio, and A549 at a 1:3 ratio.
[0108] One day before the assay, A549 and HeLa cells were seeded at 2000 cells / well in 96-well cell culture plates, and A2780 cells were seeded at 3000 cells / well in 96-well cell culture plates. 80 μL of cell suspension was seeded in each well. The cell culture plates were placed in a cell culture incubator at 37°C and 5% CO2 and incubated for 24 hours.
[0109] IV. Drug Treatment
[0110] The highest concentration of compound PCG was prepared to be 1 mM. It was gradually diluted with culture medium to achieve the appropriate concentration. After incubating the cells, the culture medium was removed, and each well was filled with prepared culture medium containing different concentrations of the test compound. Cells without the drug served as the control group, and the treated group served as the treatment group. The cells were incubated at 37°C in a 5% CO2 cell culture incubator for 24 hours before being collected.
[0111] Western blotting was used to detect the expression of STAT3, p-STAT3, BCL-2, and β-Actin proteins in cell samples.
[0112] V. Data Processing
[0113] Substitute the measured protein expression level into the following formula to calculate the relative protein expression level.
[0114] Relative protein expression level = As / Ab
[0115] As: Protein expression level in the control group / experimental group;
[0116] Ab: β-Actin protein expression level.
[0117] Use the R language to perform statistics and plot the graphs.
[0118] VI. Experimental Results
[0119] In A549, HeLa, and A2780 cells, cytosine at 0.08 μM for 24 h significantly inhibited the expression of p-STAT3, indicating that cytosine inhibited STAT3 phosphorylation, prevented p-STAT3 dimers from entering the nucleus, and thus inhibited the expression of downstream BCL-2 protein, thereby inhibiting the proliferation of tumor cells.
[0120] Experimental Example 6: Flow cytometry was used to determine the inhibitory effect of salicylic acid on the tumor cell cycle.
[0121] I. Experimental Instruments and Reagents
[0122] Instruments: Incubator: Shanghai Yiheng; Centrifuge: Sigma, Germany; Biosafety Cabinet: Haier; Flow Cytometer (FACSVerse): BD, USA.
[0123] Trial drug: Glycoside was purchased from Chengdu Yirui Biotechnology Co., Ltd.
[0124] II. Preparation of Cell Growth Culture Medium
[0125] Prepare various cell growth media under aseptic conditions, store the media in a 4°C refrigerator for later use, and preheat to 37°C before use.
[0126] III. Cell Passaging and Seeding
[0127] When the cells reached 80-90% confluence in the culture dish, they were digested with trypsin and EDTA solution, centrifuged, and then resuspended in fresh culture medium. The three cell types were then prepared into 5 × 10⁻⁶ cells / mL formulations. 4 Cell suspension of 1 cell / mL was seeded into 5 mL per well in a 60 mm culture plate. The cell plate was then incubated at 37°C in a 5% CO2 cell culture incubator for 24 hours.
[0128] IV. Drug Treatment
[0129] The highest concentration of compound PCG was prepared to be 1 mM. It was gradually diluted with culture medium to achieve the appropriate concentration. After incubating the cells, the culture medium was removed, and each well was filled with prepared culture medium containing different concentrations of the test compound. Cells without the drug served as the control group, and the treated group served as the treatment group. The cells were incubated at 37°C in a 5% CO2 cell culture incubator for 24 hours before being collected.
[0130] V. Cell Cycle Analysis
[0131] The processed cell samples were digested with trypsin solution, centrifuged, rinsed, fixed, and RNA in the cells was degraded with RNase A. After staining in the dark, the samples were then subjected to cell cycle analysis.
[0132] VI. Data Processing
[0133] Use the R language to perform statistics and plot the graphs.
[0134] VII. Experimental Results
[0135]
[0136] Table 2
[0137] The experimental results are shown in Table 2. In A549, Hela and A2780 cells, cytosine inhibited cell proliferation in the G2 / M phase, and in A549 and Hela cells, the inhibition was concentration-dependent.
[0138] References
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[0142] [4]ZHANGY, VACCARELLA S, MORGAN E, et al. Global variations in lungcancer incidence by histological subtype in 2020: population-based study[J]. Lancet Oncol, 2023, 24(11):1206-18.
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Claims
1. The use of the traditional Chinese medicine monomer, styraxoside, or its pharmaceutically acceptable salt, cocrystal, stereoisomer, prodrug, solvate, or metabolite as shown in I, in antitumor drugs.
2. The application according to claim 1, characterized in that, The tumors mentioned are selected from: pancreatic cancer, stomach cancer, breast cancer, ovarian cancer, colon cancer, cervical cancer, liver cancer, or lung cancer.
3. The application according to claim 1, characterized in that, Application of cyproterone acetate in the treatment of lung cancer, cervical cancer and ovarian cancer-related tumors.
4. The application according to claim 2 or 3, characterized in that, The half-maximal inhibitory concentration (IC50) of cytosine against A549 lung cancer cells was 0.01–0.1 μM. The half-maximal inhibitory concentration (IC50) of cytosine for HeLa cervical cancer cells was 0.01–0.2 μM; The half-maximal inhibitory concentration (IC50) of cytosine for ovarian cancer cells A2780 was 0.01–0.2 μM.
5. The application according to claim 2 or 3, characterized in that, Glycoside exerts its antitumor activity by inhibiting the phosphorylation of STAT3 protein in lung cancer cells, cervical cancer cells, and ovarian cancer cells, preventing phosphorylated STAT3 dimers from entering the cell nucleus and initiating transcription. In addition, glycoside inhibits the downstream anti-apoptotic protein BCL-2 of STAT3, synergistically inhibiting tumor cell proliferation.
6. The application according to claim 2 or 3, characterized in that, The inhibitory effect of cytosine on lung cancer cells, cervical cancer cells and ovarian cancer cells keeps cancer cells mainly in the G2 / M phase of cell proliferation.