A pharmaceutical composition for treating tumor and use thereof

Abstract

This invention relates to a pharmaceutical composition for tumor treatment and its application. Specifically, this invention relates to a composition containing a fluorouracil drug and pyrimidine nucleosides and their derivatives. Fluorouracil drugs and pyrimidine nucleotides and their derivatives have a synergistic antitumor effect; combined use can reduce the concentration of single drugs and achieve better antitumor efficacy, while reducing toxic side effects.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and more specifically, relates to an anti-tumor pharmaceutical composition. Background Technology

[0002] Hyperactive nucleotide synthesis metabolism is a common characteristic of cancer cells and an important direction for the development of anticancer drugs. Fluoropyrimidine drugs are designed based on structures similar to natural pyrimidine nucleotides. Their mechanism of action is to competitively metabolize endogenous nucleotides, disrupting the nucleotide pool homeostasis of cancer cells, interfering with and blocking the biosynthesis of DNA, RNA, and proteins, thereby exerting a therapeutic effect.

[0003] However, fluoropyrimidine drugs are metabolized and inactivated by many base-nonspecific enzymes, resulting in rapid metabolism and insufficient target specificity. Therefore, in clinical use, such drugs, like fluorouracil, require continuous intravenous infusion for 46 hours after a large intravenous bolus injection to maintain their therapeutic activity.

[0004] To overcome the problem of rapid metabolism of fluorouracil, scientists have designed drugs such as capecitabine as oral prodrugs for fluorouracil. However, these drugs cannot alter the synthetic metabolism of fluorouracil in tumor cells, and their prescribed doses still reach the gram level. Trifluuracil-tipiracil tablets are a compound fluorouracil drug that targets the same site as fluorouracil, inhibiting the synthesis of thymidine monophosphate (TMP) mediated by thymidine monophosphate synthase (TYMS). Tipiracil competitively inhibits the breakdown of trifluuracil by thymidine phosphorylase (TYMP), thereby enhancing therapeutic activity.

[0005] Although various novel molecules have been designed to modulate the metabolism of fluorouracil and its derivatives in vivo, this strategy is not only costly but also carries unpredictable side effects. Here, we propose using natural nucleotides to modulate the metabolic flux of fluorouracil drugs, thereby enhancing their therapeutic activity. Summary of the Invention

[0006] In order to address the shortcomings of existing cancer treatment drugs in clinical applications, especially the defects of most current cancer treatment drugs having large side effects and high treatment costs, this invention provides a pharmaceutical composition with a long duration of efficacy and fewer side effects.

[0007] To achieve the objectives of this invention, the inventors have provided a pharmaceutical composition for treating tumors containing effective amounts of the following active ingredients through a series of scientific formulation designs:

[0008] 1) Fluorouracil drugs and 2) Pyrimidine nucleotides and their derivatives.

[0009] Fluorouracil drugs include fluorouracil, fluorouridine, fluorouracil nucleoside, trifluthyridine, capecitabine, gemcitabine, carmoflu, tegafur, and combinations thereof.

[0010] Pyrimidine nucleotides and their derivatives, including cytosine nucleosides, cytosine nucleotides, cytosine nucleotide salts, uridine nucleosides, uridine nucleotides, uridine nucleotide salts, and combinations thereof.

[0011] In the above pharmaceutical composition, the weight ratio of fluorouracil drugs to pyrimidine nucleotides and their derivatives is 1:0.01-10000, more preferably 1:1-10000.

[0012] The pharmaceutical composition of the present invention can be used to treat various tumors in humans or animals, and its therapeutic effect on malignant tumors is also very significant. The malignant tumors include leukemia, breast cancer, lung cancer, rectal cancer, ovarian cancer, colon cancer, liver cancer, prostate cancer, stomach cancer, cervical cancer, pancreatic cancer, esophageal cancer, choriocarcinoma, malignant hydatidiform mole, bladder cancer, skin cancer, head and neck cancer, bronchogenic carcinoma, colorectal cancer, etc.

[0013] This antitumor drug composition can be administered orally or by injection for the treatment of tumors. For oral administration, the composition can be formulated into conventional oral dosage forms, such as tablets, capsules, or microemulsions. For injection administration, the composition can be formulated into conventional injectable dosage forms, such as injection solutions, sterile powders for injection, or concentrated solutions for injection. To achieve an effective therapeutic concentration in the body or at the tumor site, the appropriate administration method can be selected based on the type of tumor. For example, the drug composition of this invention can also be administered via local injection or intraperitoneal injection to obtain superior therapeutic effects. For skin tumors such as melanoma, it can also be formulated into topical preparations, including ointments, creams, or gels. Attached Figure Description

[0015] Figure 1 The results show the effects of the drug composition in Example 1 of this invention on the viability of MC38 cells.

[0016] Figure 2 The results show the effects of the drug composition in Example 1 of this invention on the viability of 4T1 cells.

[0017] Figure 3 The results show the effects of the drug composition in Example 1 of this invention on the viability of SW480 cells.

[0018] Figure 4 The results show the effects of the drug composition in Example 1 of this invention on the viability of HT29 cells.

[0019] Figure 5 This is the treatment result of the drug composition in Example 2 of the present invention on the mouse CT26 tumor model.

[0020] Detailed implementation method.

[0021] The present invention is further described in detail through specific implementation examples. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of protection of the present invention. After reading this invention, any modifications of the present invention in various equivalent forms by those skilled in the art fall within the scope defined by the appended claims. Unless otherwise specified, all raw materials and reagents of the present invention are commercially available.

[0022] Example 1

[0023] To investigate whether there is a synergistic effect between drug 1 (pyrimidine nucleotides and their derivatives) and drug 2 (fluorouracil drugs) and their impact on the viability of MC38, 4T1, SW480, and HT29 cells, the toxicity of drug 1 and drug 2 to tumor cells at different concentrations was first determined. Within the non-toxic concentration range of drug 1, gradient concentrations of drug 1 and specific concentrations of drug 2 were administered in combination, and the cells were co-incubated with tumor cells for 48-96 h. Cell viability was then assessed using CCK-8 assay.

[0024] The method for detecting cell viability is as follows: cells in the logarithmic growth phase are digested with trypsin to prepare a single-cell suspension, and cell counting is performed. The cell suspension is then divided into 3 × 10⁶ cells per well. 3 Cells were seeded at a density of [number] cells per well in 96-well cell culture plates. The seeded 96-well plates were then placed in a CO2 incubator at 37 ℃ and 5% CO2. 2 The cells were cultured for 12 hours under the specified conditions. After cell attachment, the culture medium in the wells was discarded. Subsequently, before CCK-8 cell viability assay, different concentrations of the drug were co-incubated with the cells for 48-96 hours. Next, the culture medium in the wells was discarded, and 100 μL of fresh complete cell culture medium and 10 μL of CCK-8 (Dojindo Laboratories) reagent were added. After gentle mixing, the 96-well plate was returned to the CO2 incubator for another 2 hours. Finally, the absorbance (OD value) of each well was measured using a microplate reader at wavelengths of 450 nm and 560 nm. After subtracting the background absorption at 560 nm, the obtained data were processed to calculate the cell viability and standard deviation after drug treatment. The calculation method is as follows:

[0025] Depend on Figure 1 , Figure 2 , Figure 3 and Figure 4 It is known that the combination of drug 1 and drug 2 has a synergistic effect in vitro and inhibits the viability of MC38, 4T1, SW480 and HT29 cells.

[0026] Example 2

[0027] To investigate the in vivo efficacy of the combination of drug 1 (pyrimidine nucleotides and their derivatives) and drug 2 (fluorouracil drugs), 50 μL of CT26 tumor cells were injected at a concentration of 5 × 10⁻⁶. 7 A cell suspension at a density of 100 cells / mL was subcutaneously inoculated into the right abdomen of female BALB / c mice. The day of CT26 tumor cell inoculation was designated as day 0. On day 8 post-inoculation, a group of tumors with similar volumes (approximately 100 mm²) were selected. 3 CT26 tumor-bearing mice were randomly divided into four groups of nine mice each. All mice received intraperitoneal administration three times daily for one week. Group 1 was the control group (intraperitoneal injection of saline), Group 2 was the drug 1 group (drug 1), Group 3 was the drug 2 group (drug 2), and Group 4 was the combination of drug 1 and drug 2 (drug 1 + drug 2). Starting from day 8, the mice were weighed daily, and their weight changes were recorded. Simultaneously, the major diameter (L) and minor diameter (D) of the tumor in each mouse were measured using calipers, and the formula V = L × D was applied. 2 / 2 The tumor volume of each mouse was calculated. The mean and standard error (SE) of the tumor volume for each group of mice were calculated, and tumor growth curves were plotted. On day 22, CT26 tumors were isolated from the skin of the mice and their weight was measured.

[0028] Depend on Figure 5 It can be seen that the combination of drug 1 and drug 2 has a better tumor treatment effect than drug 1 and drug 2 alone, significantly reduces tumor volume, has good safety, and does not significantly change the weight of mice.

[0029] Example 3

[0030] To investigate the in vivo efficacy of combined oral administration of drug 1 (pyrimidine nucleotides and their derivatives) and drug 2 (fluorouracil), 50 μL of CT26 tumor cells were injected at a concentration of 5 × 10⁻⁶. 7 A cell suspension at a density of 100 cells / mL was subcutaneously inoculated into the right abdomen of female BALB / c mice. The day of CT26 tumor cell inoculation was designated as day 0. These mice were randomly divided into four groups of nine mice each. All mice in each group received oral administration via gavage three times daily for one week. Group 1 was the control group (oral administration of saline), Group 2 was the drug 1 group (drug 1), Group 3 was the drug 2 group (drug 2), and Group 4 was the combination of drug 1 and drug 2 (drug 1 + drug 2). Starting from day 8, tumor volume and weight were measured daily in the mice.

[0031] The results showed that the combination of oral drugs 1 and 2 was more effective in treating tumors than drugs 1 and 2 alone, significantly reducing tumor volume and exhibiting good safety, with no significant change in mouse body weight.

[0032] Example 4

[0033] To investigate the in vivo efficacy of combined administration of drug 1 (pyrimidine nucleotides and their derivatives) and drug 2 (fluorouracil drugs) in a tumor metastasis model, 50 μL of 4T1-Luc. tumor cells were inoculated at a concentration of 5 × 10⁻⁶. 7 A cell suspension at a density of 4T1-Luc. cells / mL was inoculated into the left mammary pad of female BALB / c mice, and the day of inoculation was designated as day 0. These mice were randomly divided into four groups of nine mice each. All mice received intraperitoneal injections three times daily for one week. Group 1 was the control group (intraperitoneal injection of saline), Group 2 was the drug 1 group (drug 1), Group 3 was the drug 2 group (drug 2), and Group 4 was the combination of drug 1 and drug 2 (drug 1 + drug 2). From day 10 onwards, tumor volume and weight were measured daily. Mice were sacrificed on day 40 to observe lung tumor metastases.

[0034] The results showed that the combination of drugs 1 and 2 was more effective in treating tumors than drugs 1 and 2 alone, significantly reducing tumor volume and exhibiting good safety. The mice did not show significant changes in body weight, and the number of lung tumor metastases after treatment with the combination of drugs 1 and 2 was also less than that after treatment with drugs 1 and 2 alone.

[0035] The above descriptions are merely some embodiments of the present invention. For those skilled in the art, various modifications and improvements can be made without departing from the inventive concept of the present invention, and these all fall within the protection scope of the present invention.

Claims

1. An antitumor composition, characterized in that, The composition comprises fluorouracil drugs and pyrimidine nucleotides and their derivatives.

2. The antitumor composition according to claim 1, characterized in that, The fluorouracil drugs include at least one of fluorouracil, fluorouracil, fluorouracil nucleoside, trifluthyridine, capecitabine, gemcitabine, carmoflu, tegafur, etc.

3. The antitumor composition according to claim 1, characterized in that, The pyrimidine nucleotides and their derivatives include at least one of cytosine nucleoside, cytosine nucleotide, cytosine nucleotide salt, uridine nucleoside, uridine nucleotide, uridine nucleotide salt, etc.

4. An antitumor drug, characterized in that, It consists of the antitumor composition according to claims 1 to 3 and pharmaceutically acceptable excipients.

5. The antitumor drug according to claim 4, characterized in that, The antitumor drug is available in oral, injectable, or topical formulations.

6. The antitumor drug according to claim 5, characterized in that, The oral dosage forms are tablets, capsules, oral liquids, or granules, etc.; the injectable dosage forms are injection solutions, sterile powders for injection, or concentrated solutions for injection, etc.; the external dosage forms include ointments, creams, or gels, etc.

7. The use of the antitumor composition according to claims 1 to 3 in the preparation of an antitumor drug.

8. The application according to claim 7, characterized in that... The tumors mentioned include colon cancer, rectal cancer, breast cancer, lung cancer, liver cancer, stomach cancer, pancreatic cancer, ovarian cancer, cervical cancer, prostate cancer, bladder cancer, skin cancer, head and neck cancer, malignant hydatidiform mole, leukemia, etc.

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