Pharmaceutical compositions, pharmaceutical preparations for the treatment of breast cancer and / or ovarian cancer and use thereof

Abstract

The present application relates to the technical field of traditional Chinese medicine, and discloses a pharmaceutical composition for treating breast cancer and / or ovarian cancer, a pharmaceutical preparation and purposes thereof.The pharmaceutical composition provided by the present application comprises taxus (taxus chinensis), salvia miltiorrhiza, poria cocos and licorice, is reasonable in compatibility, conforms to the compatibility principle of "monarch, minister, assistant and guide", and can play the effects of eliminating evil sores, dispelling stasis, expelling evil, supporting healthy qi, promoting blood circulation to remove blood stasis and eliminating tumors.Experiment researches show that the pharmaceutical composition has obvious therapeutic effects on breast cancer, ovarian cancer and other cancers, is convenient to administer and has no toxicity, and provides a feasible new scheme for efficiently treating breast cancer, ovarian cancer and other cancers.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, specifically to a pharmaceutical composition, pharmaceutical preparation, and application thereof for treating breast cancer and / or ovarian cancer. Background Technology

[0002] Breast cancer and ovarian cancer are among the most common types of cancer, with a trend towards affecting younger people and a high overall mortality rate. Therefore, there is an urgent need to address and resolve the treatment issues of breast cancer and ovarian cancer.

[0003] Currently, the treatment options for breast and ovarian cancer are very limited, mainly including surgical treatment, radiotherapy, and biochemical drug treatment. However, most of these treatments are painful and have poor prognoses. Summary of the Invention

[0004] In view of this, the present invention provides a pharmaceutical composition, pharmaceutical preparation and application thereof for treating breast cancer and / or ovarian cancer, to solve the problems of painful treatment process and poor prognosis of existing breast cancer and ovarian cancer treatments.

[0005] In a first aspect, the present invention provides a pharmaceutical composition for treating breast cancer and / or ovarian cancer, wherein, by weight, the raw materials of the pharmaceutical composition comprise the following components:

[0006] Taxus chinensis 15-20 parts, Salvia miltiorrhiza 6-9 parts, Poria cocos 10-15 parts, Glycyrrhiza uralensis 3-9 parts.

[0007] In one optional embodiment, the preparation process of the pharmaceutical composition includes: Take the prescribed amount of yew raw material and perform a first subcritical pure water extraction to obtain the first extract; Take the prescribed amounts of Salvia miltiorrhiza, Poria cocos and Glycyrrhiza uralensis raw materials, mix them and perform a second subcritical pure water extraction to obtain the second extract; Mix the first extract with the second extract.

[0008] In one optional embodiment, the extraction of the prescribed amount of yew raw material using a first subcritical pure water extraction method includes: Take the prescribed amount of yew raw material and place it in purified water to moisten it, thus obtaining the moistened yew raw material. The tung oil raw material after being moistened with medicine was subjected to a first subcritical pure water extraction to obtain a first extract product. The first extract product is subjected to solid-liquid separation; The conditions for the first subcritical pure water extraction include: The extraction solvent was purified water, the solid-liquid ratio was 1:(15~30)g / mL, the extraction temperature was 150~180℃, the extraction pressure was 1.0~2.0MPa, the extraction time was 30~60min, and the stirring rate was 40~50r / min.

[0009] In one optional implementation, the conditions for the first subcritical pure water extraction include: The material-to-liquid ratio was 1:15 g / mL, the extraction temperature was 180 ℃, the extraction pressure was 1.9 MPa, the extraction time was 55 min, and the stirring rate was 40 r / min.

[0010] In one optional embodiment, the first subcritical pure water extraction is performed at least twice, and the final first extract is obtained by combining the extracts from each extraction. Solid-liquid separation of the first extract can be performed, which may involve filtration, followed by vacuum concentration as needed.

[0011] In one optional embodiment, the method of taking the prescribed amounts of Danshen, Poria cocos, and licorice raw materials, mixing them, and then performing a second subcritical pure water extraction includes: Take the prescribed amounts of Salvia miltiorrhiza, Poria cocos and Glycyrrhiza uralensis, mix them and place them in purified water to moisten the herbs, and obtain the moistened mixed raw materials. The mixed raw materials after being moistened with medicine were subjected to a second subcritical pure water extraction to obtain a second extract product; The second extract product is subjected to solid-liquid separation; The conditions for the second subcritical pure water extraction include: The extraction solvent was purified water, the solid-liquid ratio was 1:(15~30)g / mL, the extraction temperature was 120~160℃, the extraction pressure was 0.7~1.3MPa, the extraction time was 30~60min, and the stirring rate was 30~40r / min.

[0012] In one optional embodiment, the conditions for the second subcritical pure water extraction include: The material-to-liquid ratio was 1:18 g / mL, the extraction temperature was 150 ℃, the extraction pressure was 1.0 MPa, the extraction time was 60 min, and the stirring rate was 30 r / min.

[0013] In one optional embodiment, the second subcritical pure water extraction is performed at least twice, and the final second extract is obtained by combining the extracts from each extraction.

[0014] The raw materials for Taxus chinensis, Salvia miltiorrhiza, Poria cocos, and Glycyrrhiza uralensis can be raw medicinal materials or processed medicinal materials that have been crushed, cut into pieces, or sliced. The purified water used for soaking the herbs can be 20-40% of the weight of the raw materials, and the soaking time can be 1-2 hours.

[0015] In one optional embodiment, after mixing the first extract with the second extract, the resulting final extract can be concentrated to a relative density of 1.2-1.3 (60°C) for later use.

[0016] The pharmacological analysis of the above-mentioned drug composition is as follows: Taxus chinensis (yew tree), the principal medicinal herb, is cold in nature and bitter and salty in taste. It enters the lung, stomach, and liver meridians. It contains a large amount of taxanes, including paclitaxel, which inhibits the division and proliferation of tumor cells. The active ingredients in Taxus chinensis (yew tree) extract include: various taxane diterpenoids such as paclitaxel; water-soluble substances such as gibberellins, polysaccharides, tannins, and amino acids; flavonoids such as pine flavonoids and polyterpenoid ketones; and polyphenolic compounds, represented by ester-type catechins.

[0017] Danshen (Salvia miltiorrhiza), an assistant herb, is slightly cold in nature and bitter in taste. It enters the heart and liver meridians. It invigorates blood circulation, removes blood stasis, calms the mind, drains pus, and relieves pain. It treats angina pectoris, irregular menstruation, dysmenorrhea, amenorrhea, metrorrhagia, leukorrhea, abdominal masses, accumulations, abdominal pain due to blood stasis, joint pain, palpitations, insomnia, malignant sores and carbuncles, hepatosplenomegaly, and angina pectoris. The active ingredients in Danshen extract include tanshinone, salvianolic acid, cryptotanshinone, protocatechuic aldehyde, and tanshinone.

[0018] Poria cocos, used as an adjuvant herb, is neutral in nature and sweet and bland in taste. It enters the heart, spleen, lung, and kidney meridians. It has the effects of promoting diuresis and eliminating dampness, benefiting the spleen and stomach and protecting the kidneys, calming the mind and promoting the production of body fluids. The active ingredients in Poria cocos extract include polysaccharides, triterpenes, and enzymes.

[0019] Licorice, an adjuvant herb, is neutral in nature and sweet in taste. It enters the heart, lung, spleen, and stomach meridians. Its functions include tonifying the spleen and replenishing qi, clearing heat and detoxifying, resolving phlegm and relieving cough, relieving spasms and pain, and harmonizing other herbs. It is used for spleen and stomach weakness, fatigue, palpitations, shortness of breath, cough with excessive phlegm, abdominal and limb spasms and pain, carbuncles and boils, and to alleviate the toxicity and harshness of other medications. The active ingredients in licorice extract include glycyrrhizin, glycyrrhizic acid, glycyrrhizin glycosides, and glycyrrhizin flavonoids.

[0020] In a second aspect, the present invention provides a pharmaceutical preparation for treating breast cancer and / or ovarian cancer, the pharmaceutical preparation comprising the above-mentioned pharmaceutical composition.

[0021] In one optional embodiment, the dosage form of the pharmaceutical preparation includes at least one of capsules, granules, decoctions, pills, tablets, medicated wines, and oral liquids.

[0022] In one optional embodiment, the paclitaxel content in each unit dose of the pharmaceutical preparation is 1–1.5 mg. The pharmaceutical preparation is administered orally, twice daily, 1–2 units (capsules / bags) each time. It can be administered alone or in combination with chemotherapeutic drugs such as liposomal doxorubicin to inhibit tumor cell proliferation, improve patient physical response, and enhance treatment adherence.

[0023] Thirdly, the present invention provides the use of the above-mentioned pharmaceutical composition or pharmaceutical preparation in the preparation of a medicament for treating breast cancer and / or ovarian cancer.

[0024] Based on the above technical solution, the present invention has at least the following beneficial effects: (1) The pharmaceutical composition provided by this invention comprises yew (Taxus chinensis), salvia miltiorrhiza, poria cocos, and licorice. This pharmaceutical composition is rationally formulated and conforms to the principle of "principal, assistant, adjuvant, and guide" in traditional Chinese medicine, and can effectively eliminate malignant sores and swellings, disperse nodules, dispel pathogenic factors and strengthen the body's resistance, promote blood circulation and remove blood stasis, and eliminate tumors. Various experimental studies have shown that the LD50 of this pharmaceutical composition... 50 With a dosage of >30g / kg body weight and no organ toxicity after 90 days of continuous administration, and an oral bioavailability ≥20%, this drug composition is clinically applicable for the treatment of breast cancer and ovarian cancer. It can be used alone or in combination with chemotherapy drugs and can improve patients' quality of life scores by ≥28%. In other words, the pharmaceutical composition of this invention has a significant therapeutic effect on cancers such as breast cancer and ovarian cancer, and is convenient to administer without toxicity, providing a practical and feasible new solution for the efficient treatment of breast cancer, ovarian cancer, and other cancers.

[0025] (2) The pharmaceutical composition provided by the present invention is prepared by a two-step subcritical pure water extraction process of yew (Taxus chinensis), salvia miltiorrhiza, poria cocos and licorice, which is “extraction of yew alone + mixed extraction of salvia miltiorrhiza / poria cocos / licorice”. Both extractions use purified water as the sole extraction medium and no organic solvents are added. This process can achieve simultaneous and efficient enrichment of paclitaxel (extraction rate ≥0.05%), tanshinone IIA (extraction rate ≥0.032%), poria cocos polysaccharide (extraction rate ≥13.5%), and glycyrrhizic acid (extraction rate ≥2.95%). Furthermore, there are no organic solvent residues in the resulting extracts, and the total purity of the active ingredients is ≥93%, which is 12%-15% higher than that of the traditional mixed extraction mode. Moreover, due to the use of the two-step subcritical pure water extraction process, the water solubility of the pharmaceutical composition is good (≥96%), and it can be prepared into a formulation without the addition of surfactants or other adjuvants, which meets the formulation standards of the Pharmacopoeia of the People's Republic of China (2025 edition).

[0026] In other words, through the selection of components and the adjustment of the preparation process, the pharmaceutical composition of the present invention has significant effects of high efficacy, water solubility, non-toxicity, and safe and stable use.

[0027] (3) This invention has tailored a specific extraction scheme based on the characteristics of each component in the raw material, specifically: The paclitaxel content in yew (Taxus chinensis) is relatively low. It is typically extracted using organic solvents through extraction, crystallization, and recrystallization. However, paclitaxel is poorly soluble in water, and organic solvent-extracted paclitaxel requires the addition of surfactants and other adjuvants during use, which can increase side effects and affect antitumor efficacy. To address this issue, this invention employs a subcritical pure water extraction process for paclitaxel extraction from yew. However, during the development of this invention, the inventors discovered that paclitaxel in yew is a lipid-soluble component, while the active ingredients in danshen, poria cocos, and licorice are mostly water-soluble (such as poria cocos polysaccharides and glycyrrhizic acid). If a mixed extraction method is used, the lipid-soluble and water-soluble components will mutually influence each other's dissolution efficiency (dissolution competition), limiting the paclitaxel extraction rate and ultimately affecting the drug's efficacy.

[0028] To address the aforementioned issues, this invention innovatively employs a two-step subcritical pure water extraction process: "Taxus chinensis extraction alone + Salvia miltiorrhiza / Poria cocos / Glycyrrhiza uralensis mixed extraction". The extraction of Taxus chinensis alone avoids the interference of dissolution of fat-soluble components, while the mixed extraction of Salvia miltiorrhiza / Poria cocos / Glycyrrhiza uralensis allows for the synergistic dissolution of water-soluble components. This not only solves the problem of dissolution interference of mixed extraction components, but also significantly improves the dissolution rate of both fat-soluble and water-soluble components, achieving simultaneous and efficient enrichment of both fat-soluble and water-soluble components. Detailed Implementation

[0029] The following embodiments are provided to better understand the present invention and are not limited to the preferred embodiments described. They do not constitute a limitation on the content and scope of protection of the present invention. Any product that is the same as or similar to the present invention, derived by any person under the guidance of the present invention or by combining the features of the present invention with other prior art, falls within the protection scope of the present invention.

[0030] For experiments not specifically described in the examples, the procedures or conditions should be followed according to the conventional experimental procedures described in the literature in this field. Reagents or instruments whose manufacturers are not specified are all commercially available conventional reagent products.

[0031] The present invention will be further described in detail below with reference to specific embodiments, which should not be construed as limiting the scope of protection claimed by the present invention.

[0032] All medicinal materials used in the embodiments and comparative examples of this invention belong to the same batch and specification. The detection of effective components was conducted using the standard testing methods specified in the Pharmacopoeia of the People's Republic of China (2025 edition). The testing equipment and operating environment were kept consistent to ensure the objectivity and accuracy of the data comparison. The source and batch number of each medicinal material are as follows: Taxus: Sichuan Chunsheng Pharmaceutical Group Co., Ltd., batch number 230601; Danshen: Sichuan Chunsheng Pharmaceutical Group Co., Ltd., batch number 230901; Poria cocos: Sichuan Chunsheng Pharmaceutical Group Co., Ltd., batch number 230901; Licorice: Sichuan Chunsheng Pharmaceutical Group Co., Ltd., batch number 230901.

[0033] Example 1 This embodiment provides a pharmaceutical composition for treating breast cancer and / or ovarian cancer, wherein the raw materials, by weight, are: 15 parts of Taxus chinensis (yew), 6 parts of Salvia miltiorrhiza, 10 parts of Poria cocos, and 3 parts of Glycyrrhiza uralensis.

[0034] The pharmaceutical composition of this embodiment was prepared according to the following method: (1) Pretreatment: Each component of the medicinal materials is crushed into particles of about 20 mesh, and then purified water of 40% of the weight of the medicinal materials is added to each component to moisten them for 1.5 hours. (2) Taxus extraction: After the taxus raw material was moistened, it was put into a subcritical extraction vessel, and purified water was added at a material-to-liquid ratio of 1:15 (g / mL). After sealing, the equipment was started, the temperature was raised to 180℃, the pressure was adjusted to 1.9MPa, and the extraction was carried out at constant temperature and pressure for 55min. During the extraction, the mixture was stirred at a rate of 40r / min to enhance the mass transfer effect. The extraction was carried out twice and the extracts were combined to obtain the first extract. The first extract was filtered to remove the yew residue, and the filtrate was collected and concentrated under reduced pressure (65℃, vacuum degree -0.06MPa) to obtain the first extract.

[0035] (3) Mixed extraction: Take the raw materials of Danshen, Poria and Licorice after moistening, put them into the subcritical extraction vessel, add purified water at a material-to-liquid ratio of 1:18 (g / mL), seal and start the equipment, heat up to 150℃, adjust the pressure to 1.0MPa, extract at constant temperature and pressure for 60min, stir at a rate of 30r / min during the extraction, extract twice and combine the extracts to obtain the second extract; The second extract product was filtered, and the filtrate was collected to obtain the second extract.

[0036] (4) Mixing: The first extract obtained in step (2) is mixed with the second extract obtained in step (3) and concentrated to a relative density of 1.2 (60°C) to obtain the pharmaceutical composition of this embodiment.

[0037] According to the standards of the Pharmacopoeia of the People's Republic of China (2025 edition), the test results show that in the drug composition of this embodiment, the extraction rate of active ingredients of Taxus chinensis is 6.0%, the extraction rate of active ingredients of Salvia miltiorrhiza / Poria cocos / Glycyrrhiza uralensis is 15%, the total purity of active ingredients is 99%, and the water solubility of the drug composition is 95%.

[0038] Example 2 The pharmaceutical composition was prepared according to the method of Example 1, except that the raw materials involved in this example are: 18 parts of Taxus chinensis, 8 parts of Salvia miltiorrhiza, 12 parts of Poria cocos, and 6 parts of Glycyrrhiza uralensis.

[0039] Example 3 The pharmaceutical composition was prepared according to the method of Example 1, except that the raw materials involved in this example are: 20 parts of Taxus chinensis, 9 parts of Salvia miltiorrhiza, 15 parts of Poria cocos, and 9 parts of Glycyrrhiza uralensis.

[0040] Example 4 The pharmaceutical composition was prepared according to the method of Example 1, except that the conditions for subcritical pure water extraction of taxane in step (2) of this example are as follows: The material-to-liquid ratio was 1:15 g / mL, the extraction temperature was 180℃, the extraction pressure was 1.0 MPa, the extraction time was 60 min, and the stirring rate was 40 r / min.

[0041] According to the standards of the Pharmacopoeia of the People's Republic of China (2025 edition), the test results show that in the drug composition of this embodiment, the extraction rate of active ingredients of Taxus chinensis is 5.1%, the extraction rate of active ingredients of Salvia miltiorrhiza / Poria cocos / Glycyrrhiza uralensis is 14.9%, the total purity of active ingredients is 92%, and the water solubility of the drug composition is 93%.

[0042] Example 5 The pharmaceutical composition was prepared according to the method of Example 1, except that the conditions for subcritical pure water extraction of taxane in step (2) of this example are as follows: The material-to-liquid ratio was 1:30 g / mL, the extraction temperature was 150℃, the extraction pressure was 2.0 MPa, the extraction time was 30 min, and the stirring rate was 50 r / min.

[0043] According to the standards of the Pharmacopoeia of the People's Republic of China (2025 edition), the test results show that in the drug composition of this embodiment, the extraction rate of active ingredients of Taxus chinensis is 4.2%, the extraction rate of active ingredients of Salvia miltiorrhiza / Poria cocos / Glycyrrhiza uralensis is 15.1%, the total purity of active ingredients is 93%, and the water solubility of the drug composition is 96%.

[0044] Example 6 The pharmaceutical composition was prepared according to the method of Example 1. The difference is that the subcritical pure water extraction conditions of the mixed raw materials of Danshen / Fuling / Glycyrrhiza in step (3) of this example are as follows: the material-liquid ratio is 1:15 g / mL, the extraction temperature is 160℃, the extraction pressure is 0.7 MPa, the extraction time is 60 min, and the stirring rate is 30 r / min.

[0045] According to the standards of the Pharmacopoeia of the People's Republic of China (2025 edition), the test results show that in the drug composition of this embodiment, the extraction rate of active ingredients of Taxus chinensis is 6.1%, the extraction rate of active ingredients of Salvia miltiorrhiza / Poria cocos / Glycyrrhiza uralensis is 13.2%, the total purity of active ingredients is 90%, and the water solubility of the drug composition is 87%.

[0046] Example 7 The pharmaceutical composition was prepared according to the method of Example 1. The difference is that the subcritical pure water extraction conditions of the mixed raw materials of Danshen / Fuling / Glycyrrhiza in step (3) of this example are as follows: the material-liquid ratio is 1:30g / mL, the extraction temperature is 120℃, the extraction pressure is 1.3MPa, the extraction time is 30min, and the stirring rate is 40r / min.

[0047] According to the standards of the Pharmacopoeia of the People's Republic of China (2025 edition), the test results show that in the drug composition of this embodiment, the extraction rate of active ingredients of Taxus chinensis is 5.8%, the extraction rate of active ingredients of Salvia miltiorrhiza / Poria cocos / Glycyrrhiza uralensis is 13.9%, the total purity of active ingredients is 91%, and the water solubility of the drug composition is 94%.

[0048] Comparative Example 1 This comparative example provides a pharmaceutical composition, in parts by weight, comprising: 15 parts of Taxus chinensis (yew), 10 parts of Poria cocos, and 3 parts of Glycyrrhiza uralensis.

[0049] The pharmaceutical composition of this embodiment was prepared according to the following method: (1) Pretreatment: Same as step (1) in Example 1.

[0050] (2) Taxus extraction: Same as step (2) in Example 1.

[0051] (3) Mixed extraction: Take the raw materials of Poria cocos and Glycyrrhiza uralensis after moistening, put them into a subcritical extraction vessel, add purified water at a material-to-liquid ratio of 1:18 (g / mL), seal the vessel and start the equipment, heat up to 150℃, adjust the pressure to 1.0MPa, and extract at constant temperature and pressure for 60min. Stir at a rate of 30r / min during the extraction process, extract twice and combine the extracts to obtain the second extract. The second extract product was filtered, and the filtrate was collected to obtain the second extract.

[0052] (4) Mixing: The first extract obtained in step (2) is mixed with the second extract obtained in step (3) and concentrated to a relative density of 1.2 (60°C) to obtain the pharmaceutical composition of this comparative example.

[0053] Comparative Example 2 This comparative example provides a pharmaceutical composition for treating breast cancer and / or ovarian cancer, wherein the raw materials, by weight, are: 15 parts of Taxus chinensis (yew), 6 parts of Salvia miltiorrhiza, and 3 parts of Glycyrrhiza uralensis.

[0054] The pharmaceutical composition of this embodiment was prepared according to the following method: (1) Pretreatment: Same as step (1) in Example 1.

[0055] (2) Taxus extraction: Same as step (2) in Example 1.

[0056] (3) Mixed extraction: Take the raw materials of Salvia miltiorrhiza and Glycyrrhiza uralensis after moistening, put them into a subcritical extraction vessel, add purified water at a material-to-liquid ratio of 1:18 (g / mL), seal the vessel and start the equipment, heat up to 150℃, adjust the pressure to 1.0MPa, and extract at constant temperature and pressure for 60min. Stir at a rate of 30r / min during the extraction process, extract twice and combine the extracts to obtain the second extract. The second extract product was filtered, and the filtrate was collected to obtain the second extract.

[0057] (4) Mixing: The first extract obtained in step (2) is mixed with the second extract obtained in step (3) and concentrated to a relative density of 1.2 (60°C) to obtain the pharmaceutical composition of this comparative example.

[0058] Comparative Example 3 This comparative example provides a pharmaceutical composition for treating breast cancer and / or ovarian cancer, wherein the raw materials, by weight, are: 15 parts of Taxus chinensis (yew), 6 parts of Salvia miltiorrhiza, 10 parts of Poria cocos, and 3 parts of Glycyrrhiza uralensis.

[0059] The pharmaceutical composition of this comparative example was prepared according to the following method: (1) Pretreatment: Each component of the medicinal materials is crushed into particles of about 20 mesh, and then purified water of 40% of the weight of the medicinal materials is added to each component to moisten them for 1.5 hours. (2) Take the raw materials of Taxus chinensis, Salvia miltiorrhiza, Poria cocos and Glycyrrhiza uralensis after moistening, put them into a subcritical extraction vessel, add purified water at a material-to-liquid ratio of 1:15 (g / mL), seal the vessel and start the equipment, raise the temperature to 180℃, adjust the pressure to 1.9MPa, and extract at constant temperature and pressure for 55min. Stir at a rate of 40r / min during the extraction process to enhance the mass transfer effect. Extract twice and combine the extracts to obtain the extract product. The above-mentioned extract was filtered, the filtrate was collected, the extract was concentrated to a relative density of 1.2 (60°C), and the pharmaceutical composition of this comparative example was obtained.

[0060] According to the standards of the Pharmacopoeia of the People's Republic of China (2025 edition), the test results showed that in the drug composition of this comparative example, the extraction rate of active ingredients of Taxus chinensis was 4.8%, the extraction rate of active ingredients of Salvia miltiorrhiza / Poria cocos / Glycyrrhiza uralensis was 11.7%, the total purity of active ingredients was 90%, and the water solubility of the drug composition was 88%.

[0061] Comparative Example 4 This comparative example provides a pharmaceutical composition for treating breast cancer and / or ovarian cancer, wherein the raw materials, by weight, are: 15 parts of Taxus chinensis (yew), 6 parts of Salvia miltiorrhiza, 10 parts of Poria cocos, and 3 parts of Glycyrrhiza uralensis.

[0062] The pharmaceutical composition of this comparative example was prepared according to the following method: (1) Pretreatment: Each component of the medicinal materials is crushed into particles of about 20 mesh, and then purified water of 40% of the weight of the medicinal materials is added to each component to moisten them for 1.5 hours. (2) Take the raw materials of Taxus chinensis, Salvia miltiorrhiza, Poria cocos and Glycyrrhiza uralensis after moistening, put them into a subcritical extraction vessel, add purified water at a material-to-liquid ratio of 1:18 (g / mL), seal the vessel and start the equipment, heat up to 150℃, adjust the pressure to 1.0MPa, and extract at constant temperature and pressure for 60min. Stir at a rate of 30r / min during the extraction process, extract twice and combine the extracts to obtain the extract product; The above-mentioned extract was filtered, the filtrate was collected, the extract was concentrated to a relative density of 1.2 (60°C), and the pharmaceutical composition of this comparative example was obtained.

[0063] According to the standards of the Pharmacopoeia of the People's Republic of China (2025 edition), the test results showed that in the drug composition of this comparative example, the extraction rate of active ingredients of Taxus chinensis was 3.7%, the extraction rate of active ingredients of Salvia miltiorrhiza / Poria cocos / Glycyrrhiza uralensis was 12.2%, the total purity of active ingredients was 89%, and the water solubility of the drug composition was 92%.

[0064] Comparative Example 5 This comparative example provides a pharmaceutical composition for treating breast cancer and / or ovarian cancer, wherein the raw materials, by weight, are: 15 parts of Taxus chinensis (yew), 6 parts of Salvia miltiorrhiza, 10 parts of Poria cocos, and 3 parts of Glycyrrhiza uralensis.

[0065] The pharmaceutical composition of this comparative example was prepared according to the following method: (1) Pretreatment: Each component of the medicinal materials is crushed into particles of about 20 mesh, and then purified water of 40% of the weight of the medicinal materials is added to each component to moisten them for 1.5 hours. (2) Taxus extraction: After moistening the taxus raw material, put it into the extraction tank, add 75% ethanol solution at a material-to-liquid ratio of 1:15 (g / mL), reflux extract at normal pressure for 1 hour, extract twice, combine the extracts to obtain taxol extract; The above paclitaxel extract was filtered and concentrated under reduced pressure to remove alcohol, yielding the first extract.

[0066] (3) Mixed extraction: Take the raw materials of Danshen, Poria and Licorice after moistening, put them into the subcritical extraction vessel, add purified water at a material-to-liquid ratio of 1:18 (g / mL), seal and start the equipment, heat up to 150℃, adjust the pressure to 1.0MPa, extract at constant temperature and pressure for 60min, stir at a rate of 30r / min during the extraction, extract twice and combine the extracts to obtain the second extract; The second extract product was filtered, and the filtrate was collected to obtain the second extract.

[0067] (4) Mixing: The first extract obtained in step (2) is mixed with the second extract obtained in step (3) and concentrated to a relative density of 1.2 (60°C) to obtain the pharmaceutical composition of this comparative example.

[0068] According to the standards of the Pharmacopoeia of the People's Republic of China (2025 edition), the test results showed that in the drug composition of this comparative example, the extraction rate of active ingredients of Taxus chinensis was 5.2%, the extraction rate of active ingredients of Salvia miltiorrhiza / Poria cocos / Glycyrrhiza uralensis was 15%, the total purity of active ingredients was 96%, and the water solubility of the drug composition was 93%.

[0069] Experimental Example 1 This experimental example is used to verify the therapeutic effect of the pharmaceutical composition of this application on breast cancer.

[0070] 1. Establishment of a mouse model of subcutaneous xenograft tumor inoculated with MCF-7 breast cancer cells (1) Cell culture: MCF-7 cells were cultured in MEM medium containing 10% fetal bovine serum (FBS) at 37°C and 5% CO2. Cells in the logarithmic growth phase with a viability of ≥90% were selected for seeding.

[0071] (2) Animal selection Female immunodeficient mice (BALB / c-nu nude mice) aged 4–6 weeks, weighing approximately 16–18 g, were selected. Estradiol benzoate (10 mg / kg) was injected intraperitoneally every 1–2 days.

[0072] (3) Cell inoculation model The preserved cells were digested with trypsin, washed with PBS to remove residual serum, and then pipetted into a single-cell suspension, which was adjusted to a concentration of 1×10⁻⁶. 7The concentration of cells / mL was increased to obtain an MCF-7 cell suspension.

[0073] Each mouse received a subcutaneous injection of 0.1 mL (i.e., 1 × 10⁻⁶) under its right axilla. 6 (100 cells) The above-mentioned MCF-7 cell suspension. Insert the needle approximately 1 cm subcutaneously and slide it back and forth several times to prevent leakage of the cell suspension. Keep the inoculation process on ice and complete it within 30 minutes to maintain cell viability.

[0074] After vaccination, observe the injection site once a day. If nodules are observed and palpated at the injection site, and the diameter is greater than 5 mm, the model is considered successfully established; if no nodules are found, the model has failed.

[0075] After successful modeling, the tumor volume was measured every two days. When the tumor volume reached approximately (100±20) mm... 3 At that time, mice can be randomly grouped.

[0076] 2. Animal grouping and administration methods (1) Animal grouping Before modeling, measure weight, observe mental state, activity level, diet and water intake, coat color, etc.

[0077] After successful modeling, 48 mice with uniform tumor size were selected and randomly divided into 6 groups of 8 mice each: S1, S2, S3, D1, D2, and the model control group. An additional 8 normal mice that did not develop the model served as the negative control group.

[0078] (2) Dosage The dosage for each group of animals is as follows: Group S1: The drug prepared in Example 1 was administered by gavage at a dose of 0.035 g of the original drug per kg of mouse body weight, 0.2 mL each time, once a day; Group S2: The drug prepared in Example 2 was administered by gavage at a dose of 0.035 g of the original drug per kg of mouse body weight, 0.2 mL each time, once a day; Group S3: The drug prepared in Example 3 was administered by gavage at a dose of 0.035 g of the original drug per kg of mouse body weight, 0.2 mL each time, once a day; Group D1: The drug prepared in Comparative Example 1 was administered by gavage at a dose of 0.035 g of the original drug per kg of mouse body weight, 0.2 mL each time, once a day. Group D2: The drug prepared in Comparative Example 2 was administered by gavage at a dose of 0.035 g of the original drug per kg of mouse body weight, 0.2 mL each time, once a day. The model control group was given purified water by gavage, 0.2 mL each time, once a day. Negative control group: 0.2 mL of purified water was administered by gavage once daily.

[0079] (3) Dosing frequency Administration was carried out daily from 8:00 AM to 11:00 AM. Administration began after successful model establishment and grouping, and continued for 21 days. Body weight and feed consumption were measured weekly.

[0080] 3. Detection indicators (1) Tumor volume measurement During the course of treatment, tumor volume was measured and calculated weekly, as follows: The length (L) and width (W) of the tumor (ellipsoid) were measured using vernier calipers, and then the volume was calculated using an empirical formula for tumor volume. Tumor volume V = L × W 2 / 2.

[0081] (2) Tumor weight measurement and tumor inhibition rate calculation 21 days after drug administration and 24 hours after drug withdrawal, surviving mice in each group were anesthetized with isoflurane, blood was collected from the orbital vein, and then euthanized by cervical dislocation. Subcutaneous tumor tissue was excised, placed in pre-cooled calcium and magnesium-free PBS, washed away to remove surface blood, blotted dry with filter paper, accurately weighed using an analytical balance, and the tumor inhibition rate was calculated. Tumor inhibition rate (%) = (1 - average tumor weight of the current group / average tumor type of the model group) × 100%.

[0082] 4. Experimental Results (1) Tumor volume measurement results The measurement results of the subcutaneous transplanted tumor volume are shown in Table 1.

[0083] Table 1 Weekly tumor volume (mm) in mice 3 () ±SD)

[0084] The results showed that, at 7, 14, and 21 days after administration, the volume of subcutaneous tumors in mice in the S1-S3 and D1-2D groups was reduced compared with that in the model control group, and the reduction in the S1-S3 group was significantly better than that in the D1-2D group. This result indicates that, under the experimental conditions, the drug composition of this application has the effect of reducing the volume of subcutaneous xenografts of breast cancer.

[0085] (2) Tumor weight measurement results and tumor inhibition rate calculation results The results of the measurement of subcutaneous xenograft weight and the calculation of tumor inhibition rate are shown in Table 2.

[0086] Table 2. Subcutaneous xenograft weights (g) in mice of each group ( ±SD) and tumor inhibition rate (%)

[0087] The results showed that, compared with the model control group, the weight of subcutaneous tumors in mice in groups S1-S3 was reduced and the tumor inhibition rate was increased, and the effect was significantly better than that in groups D1-D2. This result indicates that, under the experimental conditions, the drug composition of this application has the effect of reducing the weight of subcutaneous xenografts of breast cancer and inhibiting tumors.

[0088] Experiment Example 2 This experimental example is used to verify the therapeutic effect of the pharmaceutical composition of this application on ovarian cancer.

[0089] 1. Establishment of a mouse model of subcutaneous xenograft tumor of SKOV3 ovarian cancer cells (1) Cell culture: SKOV3 cells were cultured in McCoy's 5A medium containing 10% fetal bovine serum (FBS) at 37°C and 5% CO2. SKOV3 cells that were passaged 3-5 times, in the logarithmic growth phase, and had a viability of ≥90% were selected for seeding.

[0090] (2) Animal selection Female immunodeficient mice (BALB / c-nu nude mice) aged 4–6 weeks, weighing approximately 16–18g, were selected.

[0091] (3) Cell inoculation model The preserved cells were digested with trypsin, washed twice with PBS to remove residual serum, and then pipetted into a single-cell suspension, adjusting the concentration to 5 × 10⁻⁶. 7 cells / mL, mixed with an equal volume of Matrigel (final concentration 2.5 × 10⁻⁶). 7 (cells / mL), handle on ice to prevent freezing.

[0092] Each mouse was subcutaneously injected with 0.2 mL of the above-mentioned SKOV3 cell suspension (containing 1×10⁻⁶ cells) under its right axilla. 6 -1×10 7 (One cell). Insert the needle about 1 cm under the skin, leave it for 10 seconds after injection, and then withdraw the needle to prevent the cell suspension from spilling out.

[0093] After vaccination, observe the injection site once a day. If nodules are observed and palpated at the injection site, and the diameter is greater than 5 mm, the model is considered successfully established; if no nodules are found, the model has failed.

[0094] After successful modeling, the tumor volume was measured every two days. When the tumor volume reached approximately (100±20) mm... 3 At that time, mice can be randomly grouped.

[0095] 2. Animal grouping and administration methods (1) Animal grouping Before modeling, measure weight, observe mental state, activity level, diet and water intake, coat color, etc.

[0096] After successful modeling, 48 mice with uniform tumor size were selected and randomly divided into 6 groups of 8 mice each: S1, S2, S3, D1, D2, and the model control group. An additional 8 normal mice that did not develop the model served as the negative control group.

[0097] (2) Dosage The dosage for each group of animals is as follows: Group S1: The drug prepared in Example 1 was administered by gavage at a dose of 0.035 g of the original drug per kg of mouse body weight, 0.2 mL each time, once a day; Group S2: The drug prepared in Example 2 was administered by gavage at a dose of 0.035 g of the original drug per kg of mouse body weight, 0.2 mL each time, once a day; Group S3: The drug prepared in Example 3 was administered by gavage at a dose of 0.035 g of the original drug per kg of mouse body weight, 0.2 mL each time, once a day; Group D1: The drug prepared in Comparative Example 1 was administered by gavage at a dose of 0.035 g of the original drug per kg of mouse body weight, 0.2 mL each time, once a day. Group D2: The drug prepared in Comparative Example 2 was administered by gavage at a dose of 0.035 g of the original drug per kg of mouse body weight, 0.2 mL each time, once a day. The model control group was given purified water by gavage, 0.2 mL each time, once a day. Negative control group: 0.2 mL of purified water was administered by gavage once daily.

[0098] (3) Dosing frequency Administration was carried out daily from 8:00 AM to 11:00 AM. Administration began after successful model establishment and grouping, and continued for 21 days. Body weight and feed consumption were measured weekly.

[0099] 3. Detection indicators (1) Tumor volume measurement During the course of treatment, tumor volume was measured and calculated weekly, as follows: The length (L) and width (W) of the tumor (ellipsoid) were measured using vernier calipers, and then the volume was calculated using an empirical formula for tumor volume. Tumor volume V = L × W 2 / 2.

[0100] (2) Tumor weight measurement and tumor inhibition rate calculation 21 days after drug administration and 24 hours after drug withdrawal, surviving mice in each group were anesthetized with isoflurane, blood was collected from the orbital vein, and then euthanized by cervical dislocation. Subcutaneous tumor tissue was excised, placed in pre-cooled calcium and magnesium-free PBS, washed away to remove surface blood, blotted dry with filter paper, accurately weighed using an analytical balance, and the tumor inhibition rate was calculated. Tumor inhibition rate (%) = (1 - average tumor weight of the current group / average tumor type of the model group) × 100%.

[0101] 4. Experimental Results (1) Tumor volume measurement results The measurement results of the subcutaneous transplanted tumor volume are shown in Table 3.

[0102] Table 3 Weekly tumor volume in mice (mm) 3 () ±SD)

[0103] The results showed that, at 7, 14, and 21 days after administration, the volume of subcutaneous tumors in mice in groups S1-S3 was significantly reduced compared to the model control group (p < 0.01); the volume of subcutaneous tumors in groups D1-D4 was also reduced, but the difference was not significant. These results indicate that, under the experimental conditions, the pharmaceutical composition of this application has the effect of reducing the volume of subcutaneous ovarian cancer xenografts.

[0104] (2) Tumor weight measurement results and tumor inhibition rate calculation results The results of the measurement of subcutaneous xenograft weight and the calculation of tumor inhibition rate are shown in Table 4.

[0105] Table 4. Subcutaneous xenograft weights (g) in mice of each group ( ±SD) and tumor inhibition rate (%)

[0106] The results showed that, compared with the model control group, the weight of subcutaneous tumors in mice in groups S1-S3 was reduced and the tumor inhibition rate was increased, both with significant differences (p < 0.01); the weight of subcutaneous tumors in mice in groups D1-D4 was also reduced and the tumor inhibition rate was also increased, but the differences were not significant. These results indicate that, under the experimental conditions, the pharmaceutical composition of this application has the effect of reducing the weight of subcutaneous ovarian cancer xenografts and inhibiting tumor growth.

[0107] Example 3 Acute toxicity test in mice 1. Test Methods (1) Animal drug administration and observation The experiment divided 110 mice into 11 groups, with 10 mice in each group, half male and half female. Groups 1-10 were the experimental groups, and group 11 was the blank control group (solvent control group). The drug concentration, dosage, and clinical human dosage ratio are shown in Table 5.

[0108] Table 5 Dosage and Grouping Table

[0109] Mice were fasted for 16 hours before oral administration, but water intake was permitted. Within 4 hours after administration, mice were closely observed and their feeding, drinking, respiration, activity, and mortality were recorded according to the items in Table 6. Observations were continued three times a day (morning, noon, and evening) for 14 consecutive days. If any animals died, observation was continued for 21 days. LD50 was calculated using the modified Kohl's method. 50 And the 95% confidence interval. The calculation formula is as follows: LD 50 =log-1[Xm-i(∑p-0.5)】; Sx50=i√∑pq / n; LD 50 95% confidence limit = log -1 (log LD) 50 +1.96 × Sx50); Note: i—group interval, i.e., the difference between the logarithmic doses of two adjacent groups; Xm—Logarithm of maximum dose; P—Morbidity rate of each dose group (mortality rates are expressed as decimals); Pm—the highest mortality rate; q—Survival rate of each dose group, q=1-p; Pn—Minimum mortality rate; ∑p—the sum of mortality rates for each dose group; n—Number of animals in each group; Sx50 — the standard error of logLD50.

[0110] Mice that died during the experiment were immediately necropsed, and any abnormal tissues or organs were examined under a histopathological microscope. Mice that did not die at the end of the observation period were euthanized and necropsed, and any abnormal tissues or organs were examined under a histopathological microscope.

[0111] When male and female laboratory animals respond significantly differently to the test drug, their test data should be compiled separately, and their respective median lethal doses (LD50) should be calculated. 50 If the sex difference is not significant, the experimental data of male and female experimental animals will be centrally collected and uniformly calculated and analyzed.

[0112] (2) Result evaluation Based on the time of onset of poisoning symptoms, symptom presentation, time of death, autopsy pathological changes, and median lethal dose (LD50) in experimental animals after drug administration. 50 (Refer to Appendix 6 for the acute toxicity (LD50) of chemicals) 50 A dose grading table is used to evaluate the investigational drug. The evaluation includes: The main symptoms of poisoning from the test substance; The median lethal dose (LD50) of the test substance 50 ) and its 95% confidence limit (mg / kg); Toxicity classification of the test substance.

[0113] 2. Results and Analysis Observe the animal's skin, fur, eyes, mucous membranes, respiratory, circulatory, central nervous system, and limb movement for 14 days after administration. If any animal dies, perform a necropsy to examine the thoracic cavity, abdominal cavity, and vital organs such as the heart, liver, spleen, lungs, and kidneys for abnormalities, bleeding points, or other abnormal morphological changes. Provide the animal's time of death distribution and LD50. 50 The calculation results and a table showing the changes in body weight of the test animals before and after the experiment are provided.

[0114] (1) Determination of LD50 after administration The results of the experimental animals 14 days after drug administration are recorded in Table 6. Since the number of dead animals, the mortality rate, and the survival rate were all 0%, the median lethal dose (LD50) of the test substance could not be calculated. 50 ) and its 95% confidence limit.

[0115] Table 6 Results of acute toxicity tests in mice after oral administration

[0116] Based on the body surface area method, the dosage for mice is approximately 12.3 times that for humans, meaning the initial dosage for mice should be 12.3 × 0.035 = 0.43 g / kg body weight. However, in this experiment, the dosage groups of 16, 32, 64, 128, 256 times, and the maximum dosage (30 g / kg body weight) exceeded the initial dosage for mice by 1.3, 2.6, 5.2, 10.4, 20.8, and 69.7 times, respectively, yet no deaths occurred within 14 days, indicating that the LD50 of the test substance is [not specified]. 50 >30 g / kg indicates a non-toxic test substance.

[0117] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A pharmaceutical composition for treating breast cancer and / or ovarian cancer, characterized in that, The raw materials of the pharmaceutical composition, expressed in parts by weight, include the following components: Taxus chinensis 15-20 parts, Salvia miltiorrhiza 6-9 parts, Poria cocos 10-15 parts, Glycyrrhiza uralensis 3-9 parts.

2. The pharmaceutical composition according to claim 1, characterized in that, The preparation process of the pharmaceutical composition includes: Take the prescribed amount of yew raw material and perform a first subcritical pure water extraction to obtain the first extract; Take the prescribed amounts of Salvia miltiorrhiza, Poria cocos and Glycyrrhiza uralensis raw materials, mix them and perform a second subcritical pure water extraction to obtain the second extract; Mix the first extract with the second extract.

3. The pharmaceutical composition according to claim 2, characterized in that, The prescribed amount of yew raw material is subjected to first subcritical pure water extraction, including: Take the prescribed amount of yew raw material and place it in purified water to moisten it, thus obtaining the moistened yew raw material. The tung oil raw material after being moistened with medicine was subjected to a first subcritical pure water extraction to obtain a first extract product. The first extract product is subjected to solid-liquid separation; The conditions for the first subcritical pure water extraction include: The extraction solvent was purified water, the solid-liquid ratio was 1:(15~30)g / mL, the extraction temperature was 150~180℃, the extraction pressure was 1.0~2.0MPa, the extraction time was 30~60min, and the stirring rate was 40~50r / min.

4. The pharmaceutical composition according to claim 3, characterized in that, The conditions for the first subcritical pure water extraction include: The material-to-liquid ratio was 1:15 g / mL, the extraction temperature was 180 ℃, the extraction pressure was 1.9 MPa, the extraction time was 55 min, and the stirring rate was 40 r / min.

5. The pharmaceutical composition according to claim 2, characterized in that, The prescribed amounts of Salvia miltiorrhiza, Poria cocos, and Glycyrrhiza uralensis are mixed and then subjected to a second subcritical pure water extraction, including: Take the prescribed amounts of Salvia miltiorrhiza, Poria cocos and Glycyrrhiza uralensis, mix them and place them in purified water to moisten the herbs, and obtain the moistened mixed raw materials. The mixed raw materials after being moistened with medicine were subjected to a second subcritical pure water extraction to obtain a second extract product; The second extract product is subjected to solid-liquid separation; The conditions for the second subcritical pure water extraction include: The extraction solvent was purified water, the solid-liquid ratio was 1:(15~30)g / mL, the extraction temperature was 120~160℃, the extraction pressure was 0.7~1.3MPa, the extraction time was 30~60min, and the stirring rate was 30~40r / min.

6. The pharmaceutical composition according to claim 5, characterized in that, The conditions for the second subcritical pure water extraction include: The material-to-liquid ratio was 1:18 g / mL, the extraction temperature was 150 ℃, the extraction pressure was 1.0 MPa, the extraction time was 60 min, and the stirring rate was 30 r / min.

7. A pharmaceutical preparation for treating breast cancer and / or ovarian cancer, characterized in that, The pharmaceutical preparation comprises the pharmaceutical composition according to any one of claims 1 to 6.

8. The pharmaceutical preparation according to claim 7, characterized in that, The dosage form of the pharmaceutical preparation includes at least one of the following: capsules, granules, decoctions, pills, tablets, medicated wines, and oral liquids.

9. The pharmaceutical preparation according to claim 7 or 8, characterized in that, In a unit dose of the pharmaceutical preparation, the content of paclitaxel is 1 to 1.5 mg.

10. The use of the pharmaceutical composition of any one of claims 1 to 6 or the pharmaceutical preparation of any one of claims 7 to 9 in the preparation of a medicament for treating breast cancer and / or ovarian cancer.

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