Pharmaceutical composition containing cordycepin and its use in the preparation of anti-tumor drugs

Abstract

The present application provides a composition containing cordycepin, which contains cordycepin and a second active ingredient selected from artemisinin, triptolide, ganoderma polysaccharide, matrine, zidovudine, acyclovir, salinomycin or DBDx. The application also provides the use of the above-mentioned composition in the preparation of anti-tumor drugs, especially for ovarian cancer, cervical cancer, lung cancer, intestinal cancer, pancreatic cancer, breast cancer and liver cancer. The present application verifies that the composition containing cordycepin has a significant inhibitory effect on the growth of various tumors and has a significant synergistic effect, which can effectively improve the treatment effect on tumors and provides a new idea for the clinical treatment of tumors.

Description

Technical Field

[0001] This invention relates to pharmaceutical compositions. Specifically, it relates to a pharmaceutical composition containing cordycepin, and further to the use of said composition in the preparation of antitumor drugs. Background Technology

[0002] Malignant tumors are a common disease that seriously threatens human health. According to statistics, in 2023, the top five cancers with the highest incidence rates in China were: lung cancer, stomach cancer, breast cancer, esophageal cancer, and liver cancer. In today's world, there is an urgent need for anti-tumor drugs.

[0003] The current strategies for developing new anti-tumor drugs include two aspects: 1. Drugs that act on tumor cells: mainly by inhibiting tumor cell proliferation, inducing tumor cell apoptosis, and inducing tumor cell differentiation. 2. Drugs that regulate the tumor microenvironment: mainly by the following aspects: (1) inhibiting tumor angiogenesis or interfering with the tumor vascular network; (2) regulating immune cells and related cytokines infiltrating the tumor; (3) regulating growth factor secretion and growth factor receptor expression; (4) inhibiting the secretion of specific enzymes and regulating corresponding inhibitory factors; (5) interfering with the transport, uptake, and excretion of substances inside and outside tumor cells. The toxicity of currently used anti-tumor drugs is a prominent problem that plagues tumor chemotherapy. Finding drugs with low toxicity that act on specific links or molecular targets in the tumor microenvironment to improve anti-tumor effects or reduce toxicity has become a new direction in anti-tumor drug research and an urgent need for clinical treatment of tumors.

[0004] Cordycepin, also known as cordycepin, is a compound with the chemical formula C0.05. 10 H 13 N5O3, with a molecular weight of 251.246, was the first nucleoside antibiotic isolated from fungi. Cordycepin is the main active ingredient of Cordyceps militaris, possessing various physiological and pharmacological activities. Existing research indicates that cordycepin exhibits protective effects on the lungs and kidneys, anti-hypertension, anti-tumor, neuroprotective, anti-inflammatory, antioxidant, and immunomodulatory activities. Therefore, cordycepin has attracted considerable attention from scholars in the fields of anti-aging, health care, and new drug development. However, current research has not reported on the combined use of cordycepin with other non-clinical antitumor drugs, nor has it verified whether such combinations have better antitumor effects. Summary of the Invention

[0005] The purpose of this invention is to provide a novel pharmaceutical composition for targeting tumor cells and regulating the tumor microenvironment. The idea behind this invention is to investigate the combined effects of cordycepin and various non-clinical antitumor drugs on the growth inhibition of various tumor cells, and to provide a novel comprehensive targeted pharmaceutical composition with significant synergistic effects.

[0006] Based on this, the present invention provides a composition containing cordycepin, the composition containing cordycepin and a second active ingredient, the second active ingredient being selected from artemisinin, triptolide, Ganoderma lucidum polysaccharide, matrine, zidovudine, acyclovir, salinomycin or DBDx.

[0007] In this invention, DBDx can be referred to the descriptions in Chinese invention patents CN 201010133844.5 and CN 201010133858.7, or to Xiujun Liu et al., A multifunctional drug combination shows highly potent therapeutic efficacy against human cancer xenografts in athymic mice. PLoS One. 2014 Dec 22;9(12):e115790. doi: 10.1371 / journal.pone.0115790. The DBDx of this invention contains dipyridamole, ubenmelast, and dexamethasone in a mass ratio of 100:20:1.

[0008] In this invention, the mass ratio of cordycepin to the second active ingredient is 0.001 to 1000:1.

[0009] According to a particularly preferred embodiment, the mass ratio of cordycepin to artemisinin is 1:1 to 1:2, the mass ratio of cordycepin to triptolide is 1:1 to 500:1, the mass ratio of cordycepin to Ganoderma lucidum polysaccharide is 1:1 to 1:2, the mass ratio of cordycepin to matrine is 1:1, the mass ratio of cordycepin to zidovudine is 1:1 to 1:5 (particularly 1:1 to 1:2), the mass ratio of cordycepin to acyclovir is 1:1, the mass ratio of cordycepin to salinomycin is 1:1 to 10:1, and the mass ratio of cordycepin to DBDx is 1:1 to 50:121 (particularly 1:1 to 10:121).

[0010] The present invention also provides pharmaceutical compositions comprising the above-described compositions.

[0011] The pharmaceutical composition of the present invention may further include a therapeutically effective amount of the aforementioned antitumor drug and a medically acceptable carrier. The medically acceptable carrier refers to pharmaceutical carriers conventional in the pharmaceutical field, such as diluents, excipients such as water, fillers such as starch and sucrose, binders such as cellulose derivatives like gelatin and polyvinylpyrrolidone, and lubricants such as talc.

[0012] Preferably, in the pharmaceutical composition, the composition containing cordycepin has a mass content of 0.5% to 99.9% based on the total mass.

[0013] The pharmaceutical complex described in the present invention can be prepared by conventional methods in the pharmaceutical field.

[0014] The present invention also provides the use of the above pharmaceutical composition in the preparation of anti-tumor drugs.

[0015] The tumors include ovarian cancer, cervical cancer, lung cancer, intestinal cancer, pancreatic cancer, breast cancer and liver cancer.

[0016] The present invention verifies that the pharmaceutical composition has a significant synergistic effect and can be used in combination to treat ovarian cancer, cervical cancer, lung cancer, intestinal cancer, pancreatic cancer, breast cancer, liver cancer, etc. Compared with single drug use, it can effectively improve the treatment effect on tumors and reduce the occurrence and development of patient complications. Detailed implementation manners

[0017] The following examples are used to non-restrictively explain the technical solutions of the present invention.

[0018] In the present invention, unless otherwise specified, "%" for explaining concentration is percentage, and ":" is mass ratio.

[0019] The present invention relates to the following drugs and reagents:

[0020] Cordycepin: standard product, purchased from Shanghai Yuanye Bio-Technology Co., Ltd.; artemisinin, triptolide, ganoderma polysaccharide, matrine, zidovudine, acyclovir, salinomycin, etc. are all raw materials, purchased from Shanghai Yuanye Bio-Technology Co., Ltd.;

[0021] The present invention relates to the following tumor cells and tumor strains:

[0022] Human ovarian cancer cells (Skov-3), human cervical cancer epithelial cells (Hela), human lung cancer cells (H460), human colon cancer cells (HT-29), human pancreatic cancer cells (PANC-1), human breast cancer cells (MCF-7) are all purchased from the Cell Center of the Institute of Basic Medicine, Chinese Academy of Medical Sciences and cultured by conventional in vitro cell culture methods. The mouse liver cancer 22 tumor strain is provided by the Tumor Laboratory of the Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and passaged in Kunming mice in vivo.

[0023] The present invention relates to the following experimental animals:

[0024] Kunming mice, female, 6-8 weeks old, weighing 18-22 g, purchased from Beijing Huafukang Bio-Technology Co., Ltd., license number SCXK(Beijing)2024-0003;

[0025] Example 1: Inhibitory effect of cordycepin-based pharmaceutical composition on different tumor cells

[0026] Human ovarian cancer cells (Skov-3), human cervical cancer epithelial cells (HeLa), human lung cancer cells (H460), human colon cancer cells (HT-29), human pancreatic cancer cells (PANC-1), and human breast cancer cells (MCF-7) were all cultured and passaged using standard methods. Cells in the logarithmic growth phase were collected, and the cell suspension concentration was adjusted to 10. 4 Approximately 180 μL / mL of cell suspension was added to each well of a 96-well plate. After incubation at 37°C for 24 h, for the experimental groups, the concentration of each component was 50 μg / mL, and for the single-component groups, the concentration was 100 μg / mL, with an addition volume of 20 μL / well. Three parallel wells were set for each group, and a blank well (containing only solvent) was included for zeroing. After incubation at 37°C for 48 h, the liquid in the 96-well plate was washed, and 20 μL of MTT (5 mg / mL) was added to each well. The plate was then incubated at 37°C for 4 h, the supernatant was discarded, and 150 μL of DMSO was added to each well. The plate was shaken for 30 min, and the cells were detected at 492 nm using a microplate reader. The inhibition rate of each composition on different cells was calculated using SPSS statistical software.

[0027]

[0028] The experimental results showed that each component had varying degrees of inhibitory effect on the six types of tumor cells used in the experiment: human ovarian cancer cells (Skov-3), human cervical cancer epithelial cells (HeLa), human lung cancer cells (H460), human colon cancer cells (HT-29), human pancreatic cancer cells (PANC-1), and human breast cancer cells (MCF-7). The inhibitory effect of each component on tumor cells was significantly higher than that of the monomer. The inhibition rate of each component on tumor cells is shown in Table 1.

[0029] Table 1. Inhibitory effect of cordycepin combined with different components on tumor cells (inhibition rate %)

[0030]

[0031] Example 2: Investigating the efficacy of a cordycepin-based pharmaceutical composition against a mouse hepatocellular carcinoma xenograft model.

[0032] Mouse ascites liver cancer cells 22 were inoculated intraperitoneally into Kunming mice. After 2-3 passages, 0.2 mL of the diluted solution was added to physiological saline at a ratio of 1:50 (v / v). mL was injected subcutaneously into one axilla of Kunming rats. Starting on day 3, the rats were divided into groups for oral administration, including a control group, cordycepin group, artemisinin group, triptolide group, Ganoderma lucidum polysaccharide group, matrine group, zidovudine group, acyclovir group, salinomycin group, DBDx group, cordycepin + artemisinin group, cordycepin + triptolide group, cordycepin + Ganoderma lucidum polysaccharide group, cordycepin + matrine group, cordycepin + zidovudine group, cordycepin + acyclovir group, cordycepin + salinomycin group, and cordycepin + DBDx group. The administration was once daily for 10 days. The control group received physiological saline. All other groups received the prescribed medications. On day 14, the animals were sacrificed, their body weight was measured, tumors were isolated and weighed, and the tumor inhibition rate and combined effect index were calculated.

[0033] The formula for calculating the Drug Combination Index (CDI) is as follows:

[0034]

[0035] The calculation is based on tumor weight. AB is the ratio of the combined drug group to the control group, and A and B are the ratios of each individual drug group to the control group. If CDI < 1, it proves that the combined effect of the two drugs is synergistic; if CDI = 1, it proves that the combined effect of the two drugs is additive; and if CDI > 1, it proves that the combined effect of the two drugs is antagonistic.

[0036] The experimental results are shown in Table 2.

[0037] Table 2. Efficacy of cordycepin-based drug compositions in a mouse hepatocellular carcinoma xenograft model.

[0038]

[0039] As shown in Table 2, the body weight of each group increased after 10 days of administration, and there was no significant difference in body weight compared with the control group. This indicates that cordycepin, each of the second components, and the cordycepin-based composition do not have a significant effect on the body weight of mice, and the combination is safe.

[0040] Table 2 shows that the cordycepin-based compositions significantly inhibited the growth of mouse hepatocellular carcinoma xenografts (HCC 22), exhibiting stronger and more significant effects than cordycepin alone and the second component. The combined use of cordycepin with the second component (artemisinin, triptolide, Ganoderma lucidum polysaccharide, matrine, zidovudine, acyclovir, salinomycin, and DBDX, among other non-clinical anticancer drugs) significantly increased the inhibition rate of mouse HCC xenografts compared to single-drug therapy, with a combined effect index of less than 1, demonstrating a significant synergistic effect. This combination effectively improves the therapeutic effect on tumors and provides a new approach for clinical tumor treatment.

Claims

1. A composition containing cordycepin, the composition containing cordycepin and a second active ingredient, the second active ingredient being selected from artemisinin, triptolide, Ganoderma lucidum polysaccharide, matrine, zidovudine, acyclovir, salinomycin or DBDx.

2. The composition containing cordycepin according to claim 1, characterized in that... The cordycepin compounds are selected from one or more of Cordyceps militaris, mycelium produced by Cordyceps militaris fermentation, and / or fermentation broth.

3. The composition containing cordycepin according to claim 1, characterized in that... The mass ratio of cordycepin to the second active ingredient is 0.001 to 1000:

1.

4. The composition containing cordycepin according to claim 1, characterized in that... The mass ratio of cordycepin to artemisinin is 1:1 to 1:2; the mass ratio of cordycepin to triptolide is 1:1 to 500:1; the mass ratio of cordycepin to Ganoderma lucidum polysaccharide is 1:1 to 1:2; the mass ratio of cordycepin to matrine is 1:1; the mass ratio of cordycepin to zidovudine is 1:1 to 1:2; the mass ratio of cordycepin to acyclovir is 1:1; the mass ratio of cordycepin to salinomycin is 1:1 to 10:1; and the mass ratio of cordycepin to DBDx is 1:1 to 50:

121.

5. A pharmaceutical composition comprising the composition according to any one of claims 1-3.

6. The pharmaceutical composition according to claim 4, characterized in that... The pharmaceutical composition contains 0.5% to 99.9% of the composition by total mass.

7. The pharmaceutical composition according to claim 4, characterized in that... The pharmaceutical composition further includes a therapeutically effective amount of an antitumor drug and / or a medically acceptable carrier.

8. Use of the composition according to claim 1 in the preparation of an antitumor drug.

9. The application according to claim 8, characterized in that... The tumors mentioned are ovarian cancer, cervical cancer, lung cancer, colorectal cancer, pancreatic cancer, breast cancer, and / or liver cancer.

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