A pharmaceutical composition for treating cervical cancer and its preparation method

Abstract

This invention belongs to the field of biomedical technology, and particularly relates to a pharmaceutical composition for treating cervical cancer and its preparation method. The pharmaceutical composition for treating cervical cancer of this invention comprises the following components: a apigenin derivative and acetylvalerin in a mass ratio of 1:(0.3~0.5). This invention obtains a apigenin derivative by introducing specific functional groups onto apigenin, which exhibits high pharmacological activity against cervical cancer; synergistically acting with acetylvalerin, it effectively kills cervical cancer cells while reducing damage to normal cells, thus providing a safer and more effective treatment option for cervical cancer patients.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and in particular relates to a pharmaceutical composition for treating cervical cancer and its preparation method. Background Technology

[0002] The development of cervical cancer is a complex process stemming from human papillomavirus (HPV) infection. In the initial stage, women are infected with high-risk HPV through sexual contact or other means. Most women can clear the virus through their own immune system, but a small percentage, due to weakened immunity or persistent viral infection, progress to precancerous lesions. Under long-term HPV stimulation, cervical epithelial cells gradually undergo atypical changes, forming cervical intraepithelial neoplasia. Low-grade lesions often resolve spontaneously, while high-grade lesions may develop into malignant tumors. Early-stage cervical cancer usually presents with no obvious symptoms, manifesting only as minimal lesions. Intervention at this stage often does not affect the patient's lifespan. If left untreated, the disease may progress to the middle or late stages, significantly increasing the difficulty of treatment. However, a cure is still possible through a combination of surgical and radiotherapy approaches.

[0003] Treatment options for cervical cancer are diverse and must be chosen based on the patient's condition and physical status. Surgical treatment is mainly for early-stage patients and can be categorized into fertility-preserving (e.g., cervical conization, radical hysterectomy) and fertility-non-preserving (e.g., extrafascial hysterectomy, radical hysterectomy, and pelvic lymph node dissection), aiming to remove tumor tissue. Radiation therapy is suitable for patients in the middle and late stages, those who are not candidates for surgery, or those requiring adjuvant therapy. It combines external beam radiation with intracavitary radiation to precisely kill cancer cells. Chemotherapy is commonly used for neoadjuvant therapy before surgery, concurrent irradiation, and palliative care in advanced stages, utilizing drugs such as bleomycin and fluorouracil to inhibit cancer cell growth. However, while chemotherapy drugs kill cancer cells, they also damage normal cells, leading to a series of side effects. These side effects not only affect the patient's quality of life but may also impact treatment efficacy.

[0004] Apigenin, a natural compound, possesses various pharmacological effects, including anti-allergic and anti-inflammatory properties, and also inhibits the growth of various cancer cells. However, its pharmacological activity against cervical cancer is relatively limited. Therefore, further research on apigenin is needed to enhance its pharmacological activity against cervical cancer by introducing specific functional groups, thus providing a safer and more effective treatment option for cervical cancer patients. Summary of the Invention

[0005] To overcome the shortcomings of existing technologies, one of the objectives of this invention is to provide a pharmaceutical composition for treating cervical cancer. The apigenin derivative in the pharmaceutical composition of this invention exhibits high pharmacological activity against cervical cancer; its synergistic effect with valerin effectively kills cancer cells while reducing damage to normal cells, thus providing a safer and more effective treatment option for cervical cancer patients.

[0006] A second objective of this invention is to provide a method for preparing a pharmaceutical composition for treating cervical cancer. This method is relatively simple and suitable for industrial production.

[0007] One of the objectives of this invention is achieved through the following technical solution:

[0008] A pharmaceutical composition for treating cervical cancer comprises the following components: apigenin derivative and acetylvalerine; wherein the mass ratio of apigenin derivative to acetylvalerine is 1:(0.3~0.5); the chemical structural formula of apigenin derivative is as follows: .

[0009] Furthermore, the preparation process of the apigenin derivative includes the following steps:

[0010] (1) Add apigenin to dichloromethane, add hydrogen peroxide and sodium hydroxide, and stir to react; the solution after reaction is extracted, dried and concentrated to obtain an intermediate;

[0011] (2) Add the intermediate and 5-(aminomethyl)pyrimidine-2,4-diol to a methanol aqueous solution, stir until homogeneous, and then heat to react. The solution after reaction is extracted, filtered, concentrated, and purified to obtain the final product.

[0012] Further, in step (1), the molar ratio of apigenin, hydrogen peroxide and sodium hydroxide is 1:(2-2.5):(1.5-1.8); the ratio of apigenin to dichloromethane is 10mmol:15-25mL; and the concentration of hydrogen peroxide is 30-40wt%.

[0013] Furthermore, the reaction in step (1) is carried out at a temperature of 0-5°C for 2-5 hours.

[0014] Further, in step (2), the molar ratio of the intermediate to 5-(aminomethyl)pyrimidine-2,4-diol is 1:(1.2-1.5); the volume ratio of the intermediate to the methanol aqueous solution is 10 mmol:30-40 mL, and the methanol aqueous solution is prepared by mixing methanol and water in a volume ratio of (3-5):1.

[0015] Furthermore, the reaction in step (2) is carried out at a temperature of 65-70°C for 3-5 hours.

[0016] The preparation method of the pharmaceutical composition for treating cervical cancer of the present invention includes the following steps:

[0017] Weigh out the prescribed amounts of acetylvalerin and apigenin derivative, mix them evenly, and add pharmaceutically acceptable excipients to obtain the final product.

[0018] Furthermore, the excipients are selected from at least one of xanthan gum, sucralose, maltodextrin, soluble starch, and lactose.

[0019] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0020] This invention prepares apigenin derivatives by introducing specific functional groups onto apigenin, which have high pharmacological activity against cervical cancer; and then works synergistically with acetylvalerin to effectively kill cancer cells while reducing damage to normal cells, thus providing a safer and more effective treatment option for cervical cancer patients. Attached Figure Description

[0021] Figure 1 The intermediate obtained in Embodiment 1 of the present invention 1 HNMR spectrum;

[0022] Figure 2 The MS spectrum of the intermediate obtained in Example 1 of this invention;

[0023] Figure 3 The apigenin derivative obtained in Example 1 of this invention 1 HNMR spectrum;

[0024] Figure 4 The MS spectrum of the apigenin derivative obtained in Example 1 of this invention is shown. Detailed Implementation

[0025] The present invention will now be further described in conjunction with specific embodiments. It should be noted that, without conflict, the various embodiments or technical features described below can be arbitrarily combined to form new embodiments. Specific conditions not specified in the embodiments shall be performed according to conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, all reagents or instruments used are conventional products obtained through commercial channels.

[0026] Example 1

[0027] A pharmaceutical composition for treating cervical cancer is prepared from apigenin derivative and acetylvalerin in a mass ratio of 1:0.4.

[0028] The preparation process of the apigenin derivative includes the following steps:

[0029]

[0030] (1) Add apigenin to dichloromethane, and slowly add hydrogen peroxide (35wt%) and sodium hydroxide at 0°C. The molar ratio of apigenin, hydrogen peroxide and sodium hydroxide is 1:2.2:1.6; the ratio of apigenin to dichloromethane is 10mmol:20mL; stir at 0°C for 3h; quench the reaction solution with water, then extract with dichloromethane, combine the organic layers, dry with anhydrous sodium sulfate, and concentrate to obtain the intermediate; the above intermediate 1 HNMR spectrum as follows Figure 1 As shown, the MS spectrum is as follows Figure 2 As shown, 1 H NMR (C 15 H 10 O6, DMSO-d6, 400 MHz) δ 12.10 (s, 1H), 10.32 (s, 1H), 9.08 (s, 1H), 7.28 (d, 2H), 6.75 (d, 2H), 6.08 (d, 1H), 5.98 (d, 1H), 4.95(s, 1H). MS (ESI) m / z: 286.05 [M]. 1 The results of HNMR and MS confirm that it is the target product.

[0031] (2) The intermediate and 5-(aminomethyl)pyrimidine-2,4-diol were added to an aqueous methanol solution, wherein the molar ratio of the intermediate to 5-(aminomethyl)pyrimidine-2,4-diol was 1:1.3; the volume ratio of the intermediate to the aqueous methanol solution was 10 mmol:30 mL, and the aqueous methanol solution was prepared by mixing methanol and water in a volume ratio of 4:1; the mixture was stirred until homogeneous, and then heated to 68°C and stirred for 4 h at this temperature; after the reaction was completed, the solvent was removed by concentration, the residue was diluted with saturated brine, and then extracted with ethyl acetate. The combined organic phases were dried with anhydrous sodium sulfate, filtered to remove the desiccant, concentrated, and subjected to column chromatography (eluent: V). 乙酸乙酯 :V 正己烷 After purification, apigenin derivatives were obtained (ratio = 10:90). Apigenin derivatives... 1 HNMR spectrum as follows Figure 3 As shown, the MS spectrum is as follows Figure 4 As shown, 1 H NMR (C 20 H 15N3O7, DMSO-d6, 400 MHz) δ 16.50 (s, 1H), 11.32(s, 1H), 11.18 (s, 1H), 10.20 (s, 1H), 9.70 (s, 2H), 7.48 (d, 2H), 7.32 (s,1H), 7.08 (s, 1H), 6.68 (d, 1H), 6.03-5.95 (m, 2H), 3.85 (s, 2H). MS (ESI) m / z: 410.10 [M+H] + . 1 The results of HNMR and MS confirm that it is the target product.

[0032] A method for preparing a pharmaceutical composition for treating cervical cancer includes the following steps: weighing the prescribed amounts of acetylvalerin and apigenin derivative, mixing them evenly, and adding xanthan gum to obtain the final product.

[0033] Example 2

[0034] A pharmaceutical composition for treating cervical cancer is prepared from apigenin derivative and acetylvalerin in a mass ratio of 1:0.3.

[0035] The preparation process of the apigenin derivative includes the following steps:

[0036] (1) Apigenin was added to dichloromethane, and hydrogen peroxide (35wt%) and sodium hydroxide were slowly added at 0°C. The molar ratio of apigenin, hydrogen peroxide and sodium hydroxide was 1:2:1.5; the ratio of apigenin to dichloromethane was 10mmol:15mL; the mixture was stirred at 0°C for 2 hours; the reaction solution was quenched with water, then extracted with dichloromethane, the organic layers were combined and dried with anhydrous sodium sulfate, and the intermediate was obtained; after... 1 H NMR and MS detection and identification showed that its structure was consistent with that of Example 1.

[0037] (2) The intermediate and 5-(aminomethyl)pyrimidine-2,4-diol were added to an aqueous methanol solution, wherein the molar ratio of the intermediate to 5-(aminomethyl)pyrimidine-2,4-diol was 1:1.2; the volume ratio of the intermediate to the aqueous methanol solution was 10 mmol:32 mL, and the aqueous methanol solution was prepared by mixing methanol and water in a volume ratio of 3:1; the mixture was stirred until homogeneous, and then heated to 65°C and stirred for 3 h at this temperature; after the reaction was completed, the solvent was removed by concentration, the residue was diluted with saturated brine, and then extracted with ethyl acetate. The combined organic phases were dried with anhydrous sodium sulfate, filtered to remove the desiccant, concentrated, and subjected to column chromatography (eluent: V). 乙酸乙酯 :V 正己烷 After purification (ratio 10:90), apigenin derivatives were obtained.1 H NMR and MS detection and identification showed that its structure was consistent with that of Example 1.

[0038] A method for preparing a pharmaceutical composition for treating cervical cancer includes the following steps: weighing the prescribed amounts of acetylvalerin and apigenin derivative, mixing them evenly, and adding sucralose to obtain the final product.

[0039] Example 3

[0040] A pharmaceutical composition for treating cervical cancer is prepared from apigenin derivative and acetylvalerin in a mass ratio of 1:0.5.

[0041] The preparation process of the apigenin derivative includes the following steps:

[0042] (1) Apigenin was added to dichloromethane, and hydrogen peroxide (40wt%) and sodium hydroxide were slowly added at 0°C. The molar ratio of apigenin, hydrogen peroxide and sodium hydroxide was 1:2.5:1.8; the ratio of apigenin to dichloromethane was 10mmol:25mL; the mixture was stirred at 5°C for 5h; the reaction solution was quenched with water, extracted with dichloromethane, the organic layers were combined and dried with anhydrous sodium sulfate, and concentrated to obtain the intermediate; after... 1 H NMR and MS detection and identification showed that its structure was consistent with that of Example 1.

[0043] (2) The intermediate and 5-(aminomethyl)pyrimidine-2,4-diol were added to an aqueous methanol solution, wherein the molar ratio of the intermediate to 5-(aminomethyl)pyrimidine-2,4-diol was 1:1.5; the volume ratio of the intermediate to the aqueous methanol solution was 10 mmol:36 mL, and the aqueous methanol solution was prepared by mixing methanol and water in a volume ratio of 5:1; the mixture was stirred until homogeneous, and then heated to 70°C and stirred for 5 h at this temperature; after the reaction was completed, the solvent was removed by concentration, the residue was diluted with saturated brine, and then extracted with ethyl acetate. The combined organic phases were dried with anhydrous sodium sulfate, filtered to remove the desiccant, concentrated, and subjected to column chromatography (eluent: V). 乙酸乙酯 :V 正己烷 After purification (ratio 10:90), apigenin derivatives were obtained. 1 H NMR and MS detection and identification showed that its structure was consistent with that of Example 1.

[0044] A method for preparing a pharmaceutical composition for treating cervical cancer includes the following steps: weighing the prescribed amounts of acetylvalerin and apigenin derivative, mixing them evenly, and adding maltodextrin to obtain the final product.

[0045] Comparative Example 1

[0046] Comparative Example 1 is basically the same as Example 1, except that the apigenin derivative is replaced with apigenin.

[0047] Comparative Example 2

[0048] Comparative Example 2 is basically the same as Example 1, except that acetylvalerin is omitted.

[0049] Experimental Example 1

[0050] (1) A apigenin derivative was dissolved in medical dimethyl sulfoxide to prepare a apigenin derivative stock solution of 20 mg / mL. The stock solution was then diluted with RPMI-1640 medium to prepare test samples of 2.5 µg / mL, 5 µg / mL, 10 µg / mL, 15 µg / mL, 20 µg / mL, 40 µg / mL, 80 µg / mL, 100 µg / mL, and 500 µg / mL. A blank control group (0 g / mL) was also set up.

[0051] SiHa cells in the logarithmic growth phase were diluted with RPMI-1640 medium containing 10% fetal bovine serum, and then cultured at a concentration of 1×10⁻⁶ cells / mL. 4 The cells were seeded at a density of 10 cells / mL into 96-well plates and incubated in a CO2 incubator for 24 h. The culture medium was then discarded, and the test samples were added, with three replicates for each concentration. Three replicates were also set up for the blank control group. For the negative control group, an equal volume of cells and RPMI-1640 medium containing 10% fetal bovine serum were added. The plates were incubated at 37°C for 72 h, and then MTT solution (5 mg / mL) was added to each well, followed by incubation at 37°C for 4 h. The supernatant was then aspirated, and DMSO was added for dissolution. The absorbance of each well was measured at 570 nm using a microplate reader to determine the inhibition rate of the sample against SiHa cells at the specified concentrations. Inhibition rate = [1 - (OD value of test sample group - OD value of blank control group) / (OD value of negative control group - OD value of blank control group)] × 100%. The IC50 of the test sample was calculated by dividing the inhibition rate by the drug concentration. 50 Values. The results are shown in Table 1.

[0052] (2) Acetylvalerin was dissolved in medical dimethyl sulfoxide to prepare a 20 mg / mL acetylvalerin stock solution. The stock solution was then diluted with RPMI-1640 medium to prepare test samples of 2.5 µg / mL, 5 µg / mL, 10 µg / mL, 15 µg / mL, 20 µg / mL, 40 µg / mL, 80 µg / mL, 100 µg / mL, and 500 µg / mL. A blank control group (0 µg / mL) and a negative control group (containing equal volumes of cells and RPMI-1640 medium with 10% fetal bovine serum) were also prepared. The SiHa cell inhibition rate of acetylvalerin was tested using the same method as for apigenin derivatives, thus determining the IC50 of acetylvalerin. 50 Values. The results are shown in Table 1.

[0053] Table 1

[0054]

[0055] As shown in Table 1, the IC50 values ​​of apigenin derivatives and acylvalerin are both less than 30 µg / mL, indicating that both have a strong inhibitory effect on the proliferation of cervical cancer cells and can be used to prepare drugs for the treatment of cervical cancer.

[0056] Experimental Example 2

[0057] (1) The pharmaceutical compositions of Examples 1-3 and Comparative Examples 1-2 were dissolved in medical dimethyl sulfoxide to prepare a stock solution of 20 mg / mL. The stock solution was then diluted with RPMI-1640 medium to prepare test samples of 2.5 µg / mL, 5 µg / mL, 10 µg / mL, 15 µg / mL, 20 µg / mL, 40 µg / mL, 80 µg / mL, 100 µg / mL, and 500 µg / mL (Examples 1-3 and Comparative Examples 1-2). A blank control group (0 µg / mL) was also set up.

[0058] (2) Take SiHa cells in the logarithmic growth phase, dilute them with RPMI-1640 medium containing 10% fetal bovine serum, and then add them at a rate of 1×10⁻⁶. 4 The cells were seeded at a density of 10 cells / mL into 96-well plates and incubated in a CO2 incubator for 24 h. The culture medium was then discarded, and the test samples were added, with three replicates for each concentration. Three replicates were also set up for the blank control group. For the negative control group, an equal volume of cells and RPMI-1640 medium containing 10% fetal bovine serum were added. The plates were incubated at 37°C for 72 h, and then MTT solution (5 mg / mL) was added to each well, followed by incubation at 37°C for 4 h. The supernatant was then aspirated, and DMSO was added for dissolution. The absorbance of each well was measured at 570 nm using a microplate reader to determine the inhibition rate of the sample against SiHa cells at the specified concentrations. Inhibition rate = [1 - (OD value of test sample group - OD value of blank control group) / (OD value of negative control group - OD value of blank control group)] × 100%. The IC50 of the test sample was calculated by dividing the inhibition rate by the drug concentration. 50 Values. The results are shown in Table 2.

[0059] Table 2

[0060]

[0061] As shown in Table 2, the half-maximal inhibitory concentration (IC50) of Examples 1-3 of the present invention is... 50The IC50 values ​​were lower than those of Comparative Examples 1-2. Compared to Example 1, Comparative Example 1 replaced the apigenin derivative with apigenin; Comparative Example 2 omitted acetylvalerin. The IC50 values ​​for both were... 50 All showed an increasing trend compared to Example 1. The above experimental results indicate that, compared to using a single apigenin derivative, the apigenin derivative and acetylvalerin in the pharmaceutical composition of the present invention can work synergistically, greatly reducing the half-maximal inhibitory concentration of the drug. Using a lower dose of the drug, it has a stronger inhibitory effect on the proliferation of cervical cancer cells.

[0062] Experimental Example 3

[0063] Normal L02 hepatocytes in logarithmic growth phase were resuspended in RPMI-1640 medium containing 10% fetal bovine serum to prepare a single-cell suspension, and 1 × 10⁻⁶ cells were cultured per well. 4 Cells were seeded at a density of 100 μL / mL into 96-well plates and cultured until cell confluence reached 90%. The old culture medium was gently removed, and normal L02 hepatocytes were co-cultured with the pharmaceutical compositions corresponding to Examples 1-3 and Comparative Examples 1-2, respectively, with the final concentration of the pharmaceutical composition in the culture medium controlled at 7 µg / mL. After culturing, MTT assay reagent was added to each well, and the cells were incubated at 37°C in the dark for 2 hours. The absorbance of each well was then measured at a wavelength of 490 nm, and cell viability was calculated. Cell viability (%) = (OD experimental group / OD blank group) × 100%. The experimental results are shown in Table 3.

[0064] Table 3

[0065]

[0066] As shown in Table 3, the pharmaceutical compositions of Examples 1-3 of this invention have no significant inhibitory effect on the activity of normal hepatocytes (LO2). This indicates that the combined use of apigenin derivatives and acetylvalerin has no significant effect on normal cells.

[0067] Compared with Example 1, Comparative Example 1 replaced apigenin derivative with apigenin, and Comparative Example 2 omitted acetylvalerin, resulting in a decrease in cell viability. This indicates that the apigenin derivative and acetylvalerin work synergistically to reduce damage to normal liver cells.

[0068] Test Example 4

[0069] Healthy female BALB / c nude mice, weighing 18-22g, were selected and acclimatized for one week in an SPF-grade laboratory at a temperature of 25℃ and a relative humidity of 50±5%, with normal food and water intake during this period. The nude mice used in this invention are hairless and lack a thymus, which prevents T cell proliferation and normal development, resulting in no transplant rejection. Therefore, they can be used for xenotransplantation and oncology research.

[0070] SiHa cells were cultured in RPMI-1640 medium (containing 10% fetal bovine serum and 1% antibiotics) under the following conditions: 37°C, 5% CO2, and saturated humidity. When the cells reached 80% confluence, they were passaged using 0.25% trypsin. The SiHa cells in the logarithmic growth phase were centrifuged and diluted with serum-free medium to a concentration of 2 × 10⁶ cells / mL. 8 After disinfection with povidone-iodine, 0.2 mL of the above cell suspension was subcutaneously injected into the right thigh root of a nude mouse under sterile conditions. The needle was then slowly withdrawn to establish a nude mouse model of SiHa cell xenograft tumor.

[0071] On the 15th day after injection, the above-mentioned cell xenograft nude mouse model was divided into Example 1 group, Example 2 group, Example 3 group, Comparative Example 1 group, Comparative Example 2 group, and model group, with 10 mice in each group. Each group was given the corresponding composition of Example 1-3 and Comparative Example 1-2 by gavage at a dose of 20 mg / kg. The model group was given an equal amount of physiological saline once a day for 14 consecutive days.

[0072] After the drug treatment was completed, nude mice were euthanized by cervical dislocation, and the tumor was completely dissected and weighed. The tumor inhibition rate was calculated as follows: Tumor inhibition rate (%) = (average tumor weight of model group - average tumor weight of experimental group) / average tumor weight of model group × 100%. The effects of the compositions of Examples 1-3 and Comparative Examples 1-2 on the tumor weight of SiHa cell transplanted tumors are shown in Table 4.

[0073] Table 4

[0074]

[0075] As can be seen from the experimental results in Table 4, the anti-cervical cancer effects of Examples 1-3 of the present invention are more obvious and the tumor inhibition rate is higher.

[0076] Compared with Example 1, Comparative Example 1 replaced apigenin derivative with apigenin; Comparative Example 2 omitted acetylvalerin. The tumor-suppressing effects of Comparative Examples 1 and 2 were not as good as those of Example 1.

[0077] The above results demonstrate that the apigenin derivative and acetylvalerin in the pharmaceutical composition of the present invention work synergistically to improve the therapeutic effect of the drug on a nude mouse model of cervical cancer while reducing the dosage of the apigenin derivative.

[0078] The above embodiments are merely preferred embodiments of the present invention and should not be construed as limiting the scope of protection of the present invention. Any non-substantial changes and substitutions made by those skilled in the art based on the present invention shall fall within the scope of protection claimed by the present invention.

Claims

1. A pharmaceutical composition for treating cervical cancer, characterized by, The application relates to a composition comprising a apigenin derivative and an acetoverine, wherein the mass ratio of the apigenin derivative and the acetoverine is 1: (0.3-0.5); the chemical structural formula of the apigenin derivative is as follows: .

2. The pharmaceutical composition for treating cervical cancer according to claim 1, wherein The preparation process of the apigenin derivative comprises the following steps: (1) adding apigenin into dichloromethane, adding hydrogen peroxide and sodium hydroxide, and stirring to react; after the reaction, the solution is extracted, dried, and concentrated to obtain an intermediate; (2) adding the intermediate and 5-(aminomethyl)pyrimidine-2,4-diol into a methanol aqueous solution, stirring uniformly, and heating to react; after the reaction, the solution is extracted, filtered, concentrated, and purified to obtain the apigenin derivative.

3. The pharmaceutical composition for treating cervical cancer according to claim 2, wherein In the step (1), the molar ratio of apigenin, hydrogen peroxide, and sodium hydroxide is 1:(2-2.5):(1.5-1.8); the dosage ratio of apigenin and dichloromethane is 10 mmol:15-25 mL; and the concentration of hydrogen peroxide is 30-40 wt%.

4. The pharmaceutical composition for treating cervical cancer according to claim 2, wherein In the step (1), the reaction temperature is 0-5°C, and the reaction time is 2-5 h.

5. The pharmaceutical composition for treating cervical cancer according to claim 2, wherein In the step (2), the molar ratio of the intermediate and 5-(aminomethyl)pyrimidine-2,4-diol is 1:(1.2-1.5); the dosage ratio of the intermediate and the methanol aqueous solution is 10 mmol:30-40 mL, and the methanol aqueous solution is prepared by mixing methanol and water in a volume ratio of (3-5):

1.

6. The pharmaceutical composition for treating cervical cancer according to claim 2, wherein In the step (2), the reaction temperature is 65-70°C, and the reaction time is 3-5 h.

7. The method of preparing a pharmaceutical composition for treating cervical cancer according to any one of claims 1 to 6, wherein, The method comprises the following steps: The formula amount of acetylvaltratum and the apigenin derivative is weighed, mixed uniformly, and then a pharmaceutically acceptable excipient is added to obtain the product.

8. The method of claim 7, wherein the pharmaceutical composition is prepared by mixing the active ingredient and the pharmaceutically acceptable carrier. The excipient is at least one selected from xanthan gum, sucralose, malt dextrin, soluble starch, and lactose.

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