Traditional Chinese medicine preparation for treating osteosarcoma and preparation process thereof

Abstract

The invention belongs to the field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine preparation for treating osteosarcoma and a preparation process thereof. The traditional Chinese medicine composition is prepared from oroxylum indicum, radix clematidis, scorpio, hedgehog hide, asarum, hot rhizoma cibotii, obscured homalomena rhizome, pseudo-ginseng, rhizoma drynariae, medulla stachyuri and kadsura pepper stem. Systematic experiments prove that the traditional Chinese medicine composition has a remarkable inhibition effect on osteosarcoma cells; in the aspect of clinical application, the prognosis condition of a patient is effectively improved through cooperation with operation and chemotherapy; and in terms of preparation performance, by optimizing the ratio of auxiliary materials and a preparation process, the obtained product is stable in quality and has important clinical application value and market potential.

Description

Technical Field

[0001] The present invention relates to the field of traditional Chinese medicine, and in particular to a traditional Chinese medicine preparation for treating osteosarcoma and a preparation process thereof. Background Art

[0002] Osteosarcoma is a high-grade, primary skeletal malignancy characterized by the deposition of immature osteoid matrix by spindle cells of mesenchymal origin. Classic osteosarcoma is one of the most common primary malignant bone tumors, with an annual incidence of 2-5 per million. It is most common in adolescents (15-25 years old). Its pathogenesis is likely related to changes induced during rapid skeletal growth, with the primary site of disease occurring in the metaphysis.

[0003] Currently, osteosarcoma treatment typically utilizes a comprehensive approach combining preoperative chemotherapy, surgery, and postoperative chemotherapy. Successful osteosarcoma treatment requires the integration of multiple techniques (surgery, chemotherapy, and radiotherapy) and the collaborative efforts of physicians from multiple disciplines (bone tumor surgery, imaging, pathology, and medical oncology).

[0004] Throughout the history of osteosarcoma treatment, before the recognition of the important role of systemic chemotherapy in controlling micrometastatic lesions, osteosarcoma was controlled through amputation. Most patients die within a year of amputation, often due to lung metastases, suggesting that patients present with lung micrometastases at the time of diagnosis. The consensus regarding osteosarcoma treatment is that amputation does not significantly improve survival in patients with osteosarcoma. While local tumor resection (limb-sparing surgery) offers significant advantages over amputation, it also carries the risk of difficulty completely removing the osteosarcoma and the possibility of recurrence.

[0005] Currently, there are few drugs other than chemotherapy drugs that are effective in the treatment of osteosarcoma, and even fewer Chinese medicines. In view of this, there is an urgent need to provide a Chinese medicine composition with significant efficacy and low recurrence rate for the auxiliary treatment of osteosarcoma. Summary of the Invention

[0006] To address the shortcomings of existing technologies, a comprehensive study of traditional Chinese medicine compositions for osteosarcoma has been conducted, resulting in a novel formulation that transcends the limitations of traditional Chinese medicine compositions for the adjuvant treatment of osteosarcoma. This invention, with a scientific and rigorous approach, has demonstrated through evidence-based pharmacological experiments that the traditional Chinese medicine composition exhibits excellent efficacy, a favorable prognostic effect, and a low recurrence rate, demonstrating significant commercial value and market potential.

[0007] Specifically, the technical solutions of the present invention are as follows: A traditional Chinese medicine composition for treating osteosarcoma is prepared from Oroxylum indicum, Clematis chinensis, Scorpion, Hedgehog peel, Asarum, Cibotium bark, Millennium Root, Panax notoginseng, Drynaria fortunei, Smilax glabra, and Piper nigrum.

[0008] The dosage of the traditional Chinese medicine composition described in the present invention is further selected, and the specific dosage of each traditional Chinese medicine is: 30-70 parts of Oroxylum indicum, 10-30 parts of Clematis chinensis, 0.5-1.5 parts of Scorpio, 1-5 parts of Hedgehog Skin, 1-3 parts of Asarum, 3-7 parts of Cibotium barometz, 3-7 parts of Millennium Jian, 8-12 parts of Panax notoginseng, 8-14 parts of Drynaria fortunei, 6-10 parts of Smilax glabra, and 5-9 parts of Piper mesenteriae.

[0009] Furthermore, the components of the traditional Chinese medicine composition are: 50 parts of Oroxylum indicum, 20 parts of Clematis chinensis, 1 part of Scorpio, 3 parts of Hedgehog Skin, 2 parts of Asarum, 5 parts of Cibotium bark, 5 parts of Millennium Jian, 10 parts of Panax notoginseng, 11 parts of Drynaria fortunei, 8 parts of Smilax glabra, and 7 parts of Piper nigrum.

[0010] Furthermore, the components of the traditional Chinese medicine composition are: 30 parts of Oroxylum indicum, 10 parts of Clematis chinensis, 0.5 parts of Scorpio, 1 part of Hedgehog Skin, 1 part of Asarum, 3 parts of Cibotium bark, 3 parts of Millennium Jian, 8 parts of Panax notoginseng, 8 parts of Drynaria fortunei, 6 parts of Smilax glabra, and 5 parts of Piper nigrum.

[0011] Furthermore, the components of the traditional Chinese medicine composition are: 70 parts of Oroxylum indicum, 30 parts of Clematis chinensis, 1.5 parts of Scorpio, 5 parts of Hedgehog Skin, 3 parts of Asarum, 7 parts of Cibotium bark, 7 parts of Millennium Jian, 12 parts of Panax notoginseng, 14 parts of Drynaria fortunei, 10 parts of Smilax glabra, and 9 parts of Piper nigrum.

[0012] Furthermore, the Chinese medicine composition of the present invention is an oral preparation; preferably, the oral preparation is a decoction, pills, tablets, mixture, capsule, granule or powder, more preferably a capsule or decoction.

[0013] The second object of the present invention is to provide a method for preparing the decoction, comprising the following steps: (1) Pre-treatment step: Grind the whole scorpion and Panax notoginseng into fine powder respectively, so that the particle size is less than 100 μm; (2) Soak the Oroxylum indicum and the Herba Lycopodii in water for 2-5 hours to extract the volatile oil, add hydroxypropyl-β-cyclodextrin to prepare an inclusion compound, and keep the residue and aqueous solution for later use; (3) Water decoction step: wash the medicinal residue and aqueous solution of step (2), asarum, hedgehog skin, scalded dog spine and drynaria root, add water and decoct, wrap the asarum with gauze and add 10-15 times of the total weight of the medicinal materials, soak for 20-40 minutes, boil over high heat, then turn to low heat and decoct for 20-40 minutes, filter, and set aside the medicinal residue and liquid; add 8-10 times of the total weight of the medicinal materials to the medicinal residue, boil over high heat, then turn to low heat and decoct for 10-30 minutes, combine the two filtrates, and set aside the filter residue; (4) Alcohol extraction step: Clematidis, Qiannianjian and Kadzu are crushed into coarse powder, and then added with the whole scorpion from step (1) and the medicinal residue from step (3) and heated under 70% ethanol for 1-2 times, each time for 1.5-3.5 hours, and the alcohol extracts are combined and distilled under reduced pressure to obtain an alcohol extract; (5) Concentration and molding: Add the filtrate from step (3) to the alcohol extract from step (4), stir and dissolve continuously, and concentrate under reduced pressure to a relative density of 1.05-1.11. Then add the fine powder of Panax notoginseng from step (1) and the inclusion compound from step (2), and stir to obtain a decoction.

[0014] Furthermore, the preparation method of the decoction is: (1) Pre-treatment step: Grind the whole scorpion and Panax notoginseng into fine powder respectively, so that the particle size is less than 100 μm; (2) Soak Oroxylum indicum and Herba Scutellariae in water for 3.5 hours to extract the volatile oil, add hydroxypropyl-β-cyclodextrin to prepare the inclusion compound, and keep the residue and aqueous solution for later use; (3) Water decoction step: wash the medicinal residue and aqueous solution of step (2), asarum, hedgehog skin, scalded dog spine and drynaria root, add water and decoct, wrap the asarum with gauze and add 13 times of the total weight of water to soak for 30 minutes, boil over high heat, then turn to low heat and decoct for 30 minutes, filter, and set aside the medicinal residue and liquid; add 9 times of the total weight of the medicinal materials to the medicinal residue, boil over high heat, then turn to low heat and decoct for 20 minutes, combine the two filtrates, and set aside the filter residue; (4) Alcohol extraction step: Clematidis, Qiannianjian and Kadzu are crushed into coarse powder, and then added with the whole scorpion from step (1) and the medicinal residue from step (3) and heated under 90% ethanol for reflux twice, each time for 3.5 hours, and the alcohol extracts are combined and distilled under reduced pressure to obtain an alcohol extract; (5) Concentration and molding: Add the filtrate from step (3) to the alcohol extract from step (4), stir and dissolve continuously, and concentrate under reduced pressure to a relative density of 1.08. Then, add the fine powder of Panax notoginseng from step (1) and the inclusion compound from step (2), and stir to obtain a decoction.

[0015] The second object of the present invention provides a method for preparing the capsule comprising the following steps: S1: Extract preparation (1) Take the Chinese medicine composition described in any one of claims 2 to 5 and set aside; (2) Pre-treatment step: Grind the whole scorpion and Panax notoginseng into fine powder respectively, and control the particle size to be less than 100 μm; (3) Soak the Oroxylum indicum and the Herba Lycopodii in water for 2-5 hours, extract the volatile oil, add hydroxypropyl-β-cyclodextrin to prepare the inclusion compound, and set aside the dregs and aqueous solution. (4) Water decoction step: add the medicinal residue and aqueous solution of step (3), asarum, hedgehog skin, scalded dog spine, and drynaria root to water and decoct; wrap the asarum with gauze and add 13 times of the total weight of water to soak for 30 minutes; boil over high heat, then turn to low heat and decoct for 30 minutes; filter and set aside the medicinal residue and liquid for later use; add 9 times of the total weight of the medicinal materials to the medicinal residue and boil over high heat, then turn to low heat and decoct for 20 minutes; combine the two filtrates and set aside the filter residue for later use; (5) Alcohol extraction step: Clematidis, Qiannianjian and Kadzu are crushed into coarse powder, and then added with the whole scorpion of step (1) and the medicinal residue of step (4) to 90% or more of ethanol and heated under reflux twice, each time for 3.5 hours, and the alcohol extracts are combined and distilled under reduced pressure to obtain an alcohol extract; (6) Concentration and molding: add the filtrate of step (4) to the alcohol extract of step (5), add ethanol to make the alcohol content 50-70%, let it stand for 10-20 hours, filter, recover the ethanol under reduced pressure and concentrate to a relative density of 1.15-1.35 to obtain a thick paste, vacuum degree -0.08MPa--0.10MPa, drying temperature 85℃-95℃, crush under vacuum state and pass through 80 mesh sieve to obtain fine powder, set aside; S2: Capsule Preparation 1) Add the fine powder to polyethylene glycol 400, mix well to dissolve, and dry at 40-60°C; add a mixture of sucrose and hydroxypropyl-β-cyclodextrin in an amount of 63-77% by weight, wherein the weight ratio of sucrose to hydroxypropyl-β-cyclodextrin in the mixture is 4-8:1, add the Panax notoginseng fine powder of step (1) in S1 and the inclusion compound of step (3), add a binder to prepare a soft material, sieve through a 16-mesh sieve to granulate, boil and dry, and set aside; 2) Add an appropriate amount of magnesium stearate to the mixture in step (1) and mix evenly. Sieve through a 16-mesh sieve to obtain dry granules, which are then filled into capsule shells to prepare capsules.

[0016] The adhesive is selected from 3% hydroxypropyl methylcellulose in 50% ethanol solution or 30% starch slurry.

[0017] Furthermore, the osteosarcoma described in the present invention is proximal femoral osteosarcoma.

[0018] The present invention has all the main, secondary, auxiliary and envoy ingredients, with appropriate compatibility and targeted mechanism. This prescription uses Oroxylum indicum and Clematidis as the main ingredients. Oroxylum indicum is cool in nature and bitter in taste, and can clear away heat and relieve sore throat, soothe the liver and soothe the stomach. In this prescription, its function of unblocking the meridians and dispersing blood stasis is utilized to promote the smooth flow of qi and blood to eliminate stasis in the bones. Clematidis is pungent, warm and amenable, and can dispel wind and dampness, dredge the meridians and relieve pain. It specializes in treating cold and dampness between the joints and is a commonly used herb for treating bone diseases. The combination of the two, one cooling and the other warming, achieves the effects of unblocking the meridians and dispersing blood stasis, dispelling pathogens and relieving pain, directly attacking the root cause of osteosarcoma. The auxiliary ingredients are scorpion, hedgehog skin, asarum and scalded dog spine to assist the main ingredient. Scorpion is good at clearing the meridians and dispelling wind, penetrating deep into bone crevices to expel pathogenic factors and enhance pain-relieving properties. Hedgehog bark is bitter, flat, and astringent, dissolving blood stasis and relieving pain, firming and stopping bleeding, aiding the main herb in dispersing blood stasis without harming vital energy. Asarum is pungent and warm, warming the meridians and dispelling cold, opening the orifices and relieving pain, drawing the medicinal power deep into the bone marrow. Scalding dog spine nourishes the liver and kidneys, strengthening the waist and knees, both promoting meridian dredging and consolidating kidney essence to nourish the body. These four herbs work together, either attacking or nourishing, strengthening the main herb's ability to dispel pathogenic factors while also protecting vital energy. The adjuvant herbs Qiannianjian, Panax notoginseng, Drynaria fortunei, and Xiaotongcao enhance synergy. Qiannianjian dispels wind and dampness, strengthens tendons and bones, and assists the principal and ministerial herbs in dredging the meridians while also nourishing tendons and bones. Panax notoginseng promotes blood circulation and dissipates blood stasis, reducing swelling and alleviating pain, dissipating bone swelling and promoting new growth. Drynaria nourishes the kidneys and strengthens bones, heals injuries and relieves pain, specifically targeting bone diseases and promoting bone tissue repair. Xiaotongcao clears heat and promotes urination, unblocks blood vessels, draws pathogenic factors out of the lower abdomen, and prevents internal accumulation of stasis and heat. Together, these four herbs serve as adjuvants, either tonic or dredging, assisting the principal and ministerial herbs in attacking pathogenic factors while regulating Qi and promoting repair. The guiding herb, Piper mesenteriae, dispels wind and dredges the meridians, promotes blood circulation and relieves pain, guiding the other herbs directly to the affected bone, harmonizing the cold and heat of the entire formula and concentrating the medicinal effects on the affected area.

[0019] Compared with the prior art, the present invention has the following advantages: The Chinese medicine composition of the present invention showed significant anti-tumor activity in pharmacological experiments, and in vitro experiments confirmed that it had a specific inhibitory effect on osteosarcoma cells Saos-2. 50 The value was significantly lower than that of the control group, and the clone formation rate was significantly reduced, indicating that it can effectively inhibit the proliferation of tumor cells.

[0020] Clinical cases have shown that this composition, as an adjuvant treatment, can significantly improve patients' tolerance to chemotherapy. No obvious recurrence was observed after combined with surgical treatment, achieving long-term survival of patients and verifying its exact clinical efficacy.

[0021] The results of pharmaceutical research show that the capsules of the present invention have excellent stability. After 6 months of accelerated testing, the water content is still less than 0.8%, the disintegration time is controlled within 16 minutes, and the retention rate of the active ingredient oroxylin exceeds 96%, which is far better than that of the control group, fully demonstrating its advantages as a modern Chinese medicine preparation. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 :IC of each example on SGC-7091 gastric cancer cells 50 antibacterial effect.

[0023] Figure 2 IC50 of each embodiment on Saos-2 osteosarcoma cells 50 bacteriostatic effect.

[0024] Figure 3 Clonogenic assay of each embodiment on SGC-7091 gastric cancer cells

[0025] Figure 4 Clonogenic assay of each embodiment on Saos-2 osteosarcoma cells

[0026] Figure 5 Appearance and detection of osteosarcoma in the upper segment of the right femur of a patient (male)

[0027] Figure 6 Overall schematic diagram of tumor-type prosthesis replacement surgery of a patient (male)

[0028] Figure 7 Detection diagram of osteosarcoma in the upper segment of the right femur of a patient (female)

[0029] Figure 8 Overall schematic diagram of tumor-type prosthesis replacement surgery of a patient (female) DETAILED DESCRIPTION

[0030] To verify the efficacy of the traditional Chinese medicine composition in the present application, the inventors carried out pharmacodynamic test research. It should be noted that the drugs selected for the pharmacodynamic test research of the present application are the drugs obtained by the representative formula and preparation method of the present application. Due to the limited space, the tests and results involved in the drugs obtained by other formulas and preparation methods included in the present application are not listed one by one.

[0031] Example 1: A Decoction for Treating Osteosarcoma and a Preparation Process Thereof Preparation process: (1) Pretreatment step: grind the scorpion and sanqi into fine powder with a particle size less than 100 μm; (2) Soak the wood butterfly and Xiaotongcao in water for 3.5 hours, extract the volatile oil, add hydroxypropyl-β-cyclodextrin to prepare the inclusion compound, and reserve the dregs and aqueous solution; (3) Water decoction step: wash the dregs and aqueous solution of step (2), fine-leafed asarum, hedgehog skin, scolopendrium, and dryopteris filix-mas, add water for decoction, wrap the fine-leafed asarum with gauze and add 13 times the total weight for soaking for 30 minutes, boil with a large fire, then switch to a small fire for decoction for 30 minutes, filter, and reserve the dregs and liquid; add 9 times the total weight of purified water to the dregs, boil with a large fire, then switch to a small fire for decoction for 20 minutes, combine the filtrates of the two times, and reserve the filter residue; (4) Alcohol extraction step: Clematidis, Qiannianjian and Kadzu are crushed into coarse powder, and then added with the whole scorpion from step (1) and the medicinal residue from step (3) and heated under 90% ethanol for reflux twice, each time for 3.5 hours, and the alcohol extracts are combined and distilled under reduced pressure to obtain an alcohol extract; (5) Concentration and molding: Add the filtrate from step (3) to the alcohol extract from step (4), stir and dissolve continuously, and concentrate under reduced pressure to a relative density of 1.08. Then, add the fine powder of Panax notoginseng from step (1) and the inclusion compound from step (2), and stir to obtain a decoction.

[0032] Example 2: A decoction for treating osteosarcoma and its preparation process Preparation process: (1) Pre-treatment step: Grind the whole scorpion and Panax notoginseng into fine powder respectively, so that the particle size is less than 100 μm; (2) Soak Oroxylum indicum and Herba Scutellariae in water for 2 hours to extract the volatile oil, add hydroxypropyl-β-cyclodextrin to prepare the inclusion compound, and keep the residue and aqueous solution for later use; (3) Water decoction step: wash the medicinal residue and aqueous solution of step (2), asarum, hedgehog skin, scalded dog spine and drynaria root, add water and decoct, wrap the asarum with gauze and add 10 times of the total weight of water to soak for 20 minutes, boil over high heat, then turn to low heat and decoct for 20 minutes, filter, and set aside the medicinal residue and liquid; add 8 times of the total weight of the medicinal materials to the medicinal residue, boil over high heat, then turn to low heat and decoct for 10 minutes, combine the two filtrates, and set aside the filter residue; (3) Alcohol extraction step: Clematidis, Qiannianjian and Kadzu are crushed into coarse powder, and then added with 70% or more ethanol, the whole scorpion from step (1) and the medicinal residue from step (3), and heated under reflux for 1.5 hours each time. The alcohol extracts are combined and vacuum distilled to obtain an alcohol extract; (4) Concentration and molding: Add the filtrate from step (2) to the alcohol extract from step (3), stir and dissolve continuously, concentrate under reduced pressure to a relative density of 1.05, then add the fine powder of Panax notoginseng from step (1) and the inclusion compound from step (2), stir and obtain a decoction.

[0033] Example 3: A decoction for treating osteosarcoma and its preparation process Preparation process: (1) Pre-treatment step: Grind the whole scorpion and Panax notoginseng into fine powder respectively, so that the particle size is less than 100 μm; (2) Soak Oroxylum indicum and Herba Scutellariae in water for 5 hours to extract the volatile oil, add hydroxypropyl-β-cyclodextrin to prepare the inclusion compound, and keep the residue and aqueous solution for later use; (3) Water decoction step: wash the medicinal residue and aqueous solution of step (2), asarum, hedgehog skin, scalded dog spine and drynaria root, add water and decoct, wrap the asarum with gauze and add 15 times of the total weight of water to soak for 40 minutes, boil over high heat, then turn to low heat and decoct for 40 minutes, filter, and set aside the medicinal residue and liquid; add 8-10 times of the total weight of the medicinal materials to the medicinal residue, boil over high heat, then turn to low heat and decoct for 30 minutes, combine the two filtrates, and set aside the filter residue; (4) Alcohol extraction step: Clematidis, Qiannianjian and Kadzu are crushed into coarse powder, and then added with 70% or more ethanol and heated under reflux twice, each time for 3.5 hours, and the alcohol extracts are combined and distilled under reduced pressure to obtain an alcohol extract; (5) Concentration and molding: Add the filtrate from step (2) to the alcohol extract from step (3), stir and dissolve continuously, concentrate under reduced pressure to a relative density of 1.11, then add the fine powder of Panax notoginseng from step (1) and the inclusion compound from step (2), stir and obtain a decoction.

[0034] Example 4: A capsule for treating osteosarcoma and its preparation process S1: Extract preparation (1) Take the Chinese medicinal materials of Example 1 and carry out the following preparation process; (2) Pre-treatment step: Grind the whole scorpion and Panax notoginseng into fine powder respectively, and control the particle size to be less than 100 μm; (3) Soak the Oroxylum indicum and the Herba Lycopodii in water for 2-5 hours, extract the volatile oil, add hydroxypropyl-β-cyclodextrin to prepare the inclusion compound, and set aside the dregs and aqueous solution. (4) Water decoction step: add the medicinal residue and aqueous solution of step (3), asarum, hedgehog skin, scalded dog spine, and drynaria root to water and decoct; wrap the asarum with gauze and add 13 times of the total weight of water to soak for 30 minutes; boil over high heat, then turn to low heat and decoct for 30 minutes; filter and set aside the medicinal residue and liquid for later use; add 9 times of the total weight of the medicinal materials to the medicinal residue and boil over high heat, then turn to low heat and decoct for 20 minutes; combine the two filtrates and set aside the filter residue for later use; (5) Alcohol extraction step: Clematidis, Qiannianjian and Kadzu are crushed into coarse powder, and then added with the whole scorpion of step (1) and the medicinal residue of step (4) to 90% or more of ethanol and heated under reflux twice, each time for 3.5 hours, and the alcohol extracts are combined and distilled under reduced pressure to obtain an alcohol extract; (6) Concentration and molding: add the filtrate of step (4) to the alcohol extract of step (5), add ethanol to make the alcohol content 50-70%, let it stand for 10-20 hours, filter, recover the ethanol under reduced pressure and concentrate to a relative density of 1.15-1.35 to obtain a thick paste, vacuum degree -0.08MPa--0.10MPa, drying temperature 85℃-95℃, crush under vacuum state and pass through 80 mesh sieve to obtain fine powder, set aside; S2: Capsule Preparation 1) Add the fine powder to polyethylene glycol 400, mix well to dissolve, and dry at 40-60°C; add a mixture of sucrose and hydroxypropyl-β-cyclodextrin in an amount of 63% by weight, wherein the weight ratio of sucrose to hydroxypropyl-β-cyclodextrin in the mixture is 4:1; add the Panax notoginseng fine powder of step (1) in S1 and the inclusion compound of step (3); add a 50% ethanol solution of 3% hydroxypropyl methylcellulose to prepare a soft material; granulate through a 16-mesh sieve, boil and dry, and set aside; 2) Add an appropriate amount of magnesium stearate to the mixture in step (1) and mix evenly. Sieve through a 16-mesh sieve to obtain dry granules, which are then filled into capsule shells to prepare capsules.

[0035] Example 5: A capsule for treating osteosarcoma and its preparation process S1: Same as Example 4 S2: Capsule Preparation 1) Add the fine powder to polyethylene glycol 400, mix well to dissolve, and dry at 40-60°C; add a mixture of sucrose and hydroxypropyl-β-cyclodextrin in an amount of 77% by weight, wherein the weight ratio of sucrose to hydroxypropyl-β-cyclodextrin in the mixture is 8:1, add the Panax notoginseng fine powder of step (1) in S1 and the inclusion compound of step (3), add 3% hydroxypropyl methylcellulose in 50% ethanol to prepare a soft material, sieve through a 16-mesh sieve to granulate, boil and dry, and set aside; 2) Add an appropriate amount of magnesium stearate to the mixture in step (1) and mix evenly. Sieve through a 16-mesh sieve to obtain dry granules, which are then filled into capsule shells to prepare capsules.

[0036] Example 6: A capsule for treating osteosarcoma and its preparation process S1: Same as Example 4 S2: Capsule Preparation 1) Add the fine powder to polyethylene glycol 400, mix well to dissolve, and dry at 40-60°C; add a mixture of sucrose and hydroxypropyl-β-cyclodextrin in an amount of 70% by weight, wherein the weight ratio of sucrose to hydroxypropyl-β-cyclodextrin in the mixture is 6:1, add the Panax notoginseng fine powder of step (1) in S1 and the inclusion compound of step (3), add an appropriate amount of 30% starch slurry to prepare a soft material, sieve through a 16-mesh sieve to granulate, boil and dry, and set aside; 2) Add an appropriate amount of magnesium stearate to the mixture in step (1) and mix evenly. Sieve through a 16-mesh sieve to obtain dry granules, which are then filled into capsule shells to prepare capsules.

[0037] Comparative Example 1: A decoction for treating osteosarcoma and its preparation process The preparation process is the same as that described in Example 1.

[0038] Comparative Example 2: A decoction for treating osteosarcoma and its preparation process The preparation process is the same as that described in Example 1.

[0039] Comparative Example 3: A decoction for treating osteosarcoma and its preparation process The preparation process is the same as that described in Example 1.

[0040] Comparative Example 4: A decoction for treating osteosarcoma and its preparation process The preparation process is the same as that described in Example 1.

[0041] Comparative Example 5: A decoction for treating osteosarcoma and its preparation process The preparation process is the same as that described in Example 1.

[0042] Comparative Example 6: A decoction for treating osteosarcoma and its preparation process The preparation process is the same as that described in Example 1.

[0043] Comparative Example 7: A capsule for treating osteosarcoma and its preparation process S1: Same as Example 4 S2: Capsule Preparation 1) Add the fine powder to a mixture of 70% by weight of sucrose and hydroxypropyl-β-cyclodextrin, wherein the weight ratio of sucrose to hydroxypropyl-β-cyclodextrin in the mixture is 6:1, add the Panax notoginseng fine powder of step (1) and the inclusion compound of step (3), add an appropriate amount of 30% starch slurry to prepare a soft material, sieve through a 16-mesh sieve to granulate, boil and dry, and set aside; 2) Add an appropriate amount of magnesium stearate to the mixture in step (1) and mix evenly. Sieve through a 16-mesh sieve to obtain dry granules, which are then filled into capsule shells to prepare capsules.

[0044] Comparative Example 8: A capsule for treating osteosarcoma and its preparation process S1: Same as Example 4 S2: Capsule Preparation 1) Add the fine powder to polyethylene glycol 400, mix well and dissolve, and dry at 40-60°C; add 70% by weight of sucrose, add the Panax notoginseng fine powder of step (1) in S1 and the inclusion compound of step (3), add an appropriate amount of 30% starch slurry to prepare a soft material, sieve through a 16-mesh sieve to granulate, boil and dry, and set aside; 2) Add an appropriate amount of magnesium stearate to the mixture in step (1) and mix evenly. Sieve through a 16-mesh sieve to obtain dry granules, which are then filled into capsule shells to prepare capsules.

[0045] Comparative Example 9: A capsule for treating osteosarcoma and its preparation process S1: Same as Example 4 S2: Capsule Preparation 1) Add the fine powder to polyethylene glycol 400, mix well to dissolve, and dry at 40-60°C; add a mixture of sucrose and hydroxypropyl-β-cyclodextrin in an amount of 70% by weight, wherein the weight ratio of sucrose to hydroxypropyl-β-cyclodextrin in the mixture is 1:6, add the Panax notoginseng fine powder of step (1) in S1 and the inclusion compound of step (3), add an appropriate amount of 30% starch slurry to prepare a soft material, granulate through a 16-mesh sieve, boil and dry, and set aside; 2) Add an appropriate amount of magnesium stearate to the mixture in step (1) and mix evenly. Sieve through a 16-mesh sieve to obtain dry granules, which are then filled into capsule shells to prepare capsules.

[0046] 1. Experimental verification of the effect of the Chinese medicine composition of the present invention 1. Cell selection and drugs Human gastric cancer cells SGC-7901 and human osteosarcoma cells Saos-2 were subcultured, and the decoctions of Examples 1-3 and Comparative Examples 1-6 were used.

[0047] 2. Experimental Methods 2.1 Monitoring the inhibitory effect on human gastric cancer cells SGC-7901 and human osteosarcoma cells Saos-2 Cells in the logarithmic growth phase were used for the experiment. The cells were digested, counted, and prepared into a cell suspension. The suspension was then seeded into a 96-well plate (100 μL / well) and incubated in a 37°C, 5% CO2 incubator for 24 hours. The test substance at the corresponding concentration was added to each well. A negative control group and a blank group were also set up, with 5 replicates per group. After the plate was incubated in the incubator for 72 hours, the cell morphology of each group was observed under a microscope. 10 μL of CCK8 solution was added to each well and incubated in the cell incubator for another 4 hours. The absorbance was measured at 450 nm, and the IC was calculated. 50 .

[0048] 2.2 Cloning efficiency of human gastric cancer cell line SGC-7901 and human osteosarcoma cell line Saos-2 For the clone formation assay, SGC-7091 and SAO-2 cells were seeded in 96-well plates at a density of 800 cells / well and cultured in an incubator. The blank group, Examples 1-3, and Comparative Examples 1-6 were then cultured for 14 days and stained with crystal violet for 10 minutes. The number of cell colonies was counted under an inverted fluorescence microscope (a colony containing 50 or more cells) and the clone formation rate was calculated.

[0049] 3. Statistical processing The data are expressed as mean ± SD. GraphPad Prism 9.0 statistical software was used. Tests were used for comparisons between two groups, and one-way Anova (Dunnett) was used for comparisons between multiple groups. P < 0.05 was considered statistically significant.

[0050] 4. Results Analysis 4.1 Analysis of the inhibitory effect on human gastric cancer cells SGC-7901 and human osteosarcoma cells Saos-2 Figure 1 :IC of each example on SGC-7091 gastric cancer cells 50 The antibacterial effect of Example 1-3 is significant. 50 The concentration of the present invention is significantly reduced, which further illustrates that the dosage required for achieving the same effect is lower than that of other types of combinations.

[0051] Figure 2 : The IC of Saos-2 osteosarcoma cells in each example 50 The ## indicates a significant difference (P<0.05). Compared with SGC-7091 gastric cancer cells, the effect is more significant, indicating that the inhibitory effect on osteosarcoma cells is more significant and prominent.

[0052] 4.2 Analysis of clone formation experiment results Figure 3 : Colony formation experiments on SGC-7091 gastric cancer cells in each example. ## indicates a significant difference (P<0.05). The colony formation rates of gastric cancer cells in Examples 1-3 were significantly lower than those in Comparative Examples 1-6. There was no statistically significant difference in colony formation rates between Comparative Examples 1-6.

[0053] Figure 4 : Colony formation experiments on Saos-2 osteosarcoma cells in each example. ## indicates significant difference (P<0.05). The osteosarcoma cell colony formation rates in Examples 1-3 were significantly lower than those in Comparative Examples 1-6. There was no statistically significant difference in the osteosarcoma cell colony formation rates between Comparative Examples 1-6.

[0054] 2. Clinical Cases Typical Case 1. Name: Liu ***, male, 17 years old, admitted to hospital on February 14, 2018 Chief complaint: A gradually enlarging mass was found in the proximal right thigh for 2 months and pain for half a month Physical examination: A palm-sized hard mass with clear boundaries, fixed and immobile, tenderness, high skin temperature, normal skin color, no venous distension, positive right 4-number test, no obvious limitation of right hip joint movement, and no other significant abnormalities were found during the rest of the physical examination.

[0055] Diagnosis: osteosarcoma of the right proximal femur (pathological diagnosis of osteosarcoma by puncture), such as Figure 5 shown.

[0056] Treatment course: DIA chemotherapy regimen (consisting of cisplatin (DDP), ifosfamide (IFO) and doxorubicin (ADM)) was performed in sequence.

[0057] The order of medication is: first give DDP chemotherapy, then give IFO and ADM after a one-week interval, which is one course of treatment, and give the second course of chemotherapy before surgery after a two-week interval. ), give the Chinese medicine decoction of Example 3 orally for one week before surgery, 100 mL each time, twice a day, and then undergo surgical treatment, tumor segment resection, tumor-type prosthesis replacement ( Figure 6 After surgery, the patient received 8 chemotherapy treatments in sequence and was given the Chinese herbal decoction of Example 3 orally, 100 mL each time, twice a day. The patient is still available for follow-up and in contact with WeChat, and is surviving long-term.

[0058] Case 2: Name: Zheng ***, female, 23 years old; admitted to hospital on January 16, 2022 Chief complaint: A gradually enlarging mass was found in the proximal right thigh for one month, which was painful when pressed and had a fever of 37.5℃. Physical examination: A palm-sized hard mass with clear boundaries, fixed and immobile, tenderness, high skin temperature, normal skin color, no venous distension, positive right 4-number test, no obvious limitation of right hip joint movement, and no other significant abnormalities were found during the rest of the physical examination.

[0059] The patient had a palpable hard mass in the anterolateral proximal part of the right thigh. Combined with its clear boundaries, fixation, tenderness, high skin temperature, and positive four-digit test, it was initially considered to be a bone tumor or bone tumor-like lesion.

[0060] Diagnosis: osteosarcoma of the right proximal femur (pathological diagnosis of osteosarcoma by puncture), such as Figure 7 shown.

[0061] Treatment course: DIA chemotherapy regimen (consisting of cisplatin (DDP), ifosfamide (IFO) and doxorubicin (ADM)) was performed in sequence.

[0062] The order of medication was: DDP chemotherapy, followed by an interval of 1 week; then IFO and ADM for 1 course of treatment, followed by a 2-week interval before the second course of chemotherapy). The Chinese medicine decoction of Example 3 was administered orally for one week before the operation, 100 mL each time, twice a day (the patient had poor tolerance), and then surgical treatment was performed, including tumor segment resection and tumor-type prosthesis replacement ( Figure 8 After the operation, the patient received three chemotherapy sessions in sequence and was given the capsules of Example 4, three times a day, four capsules each time, and is still alive.

[0063] 3. Determination of physical and chemical components of capsules Accelerated testing: Place the product at a temperature of 40°C ± 2°C and a relative humidity of 70% ± 5% for 6 months, and then conduct performance testing.

[0064] 3.1 Tests for dispersion uniformity, disintegration time, water content, and long-term testing Method for determining water content: The water content of the capsule is calculated using the drying method.

[0065] Method for testing dispersion uniformity: Take 2 capsules each of the capsules prepared in Examples 4-6 and Comparative Examples 7-9, place the inner particles in 100 mL of water at 20°C ± 1°C, shake for 3 minutes, and pass through a No. 2 sieve.

[0066] Disintegration time detection method: 6 capsules of each of Examples 1-6 and Comparative Examples 7-9 were taken and tested in water at a temperature of 37°C ± 1°C using a lifting tablet disintegration instrument.

[0067] The test results are shown in Table 1 Table 1 Test results of Examples 4-6 and Comparative Examples 7-9 The accelerated test results in Table 1 show that Comparative Examples 7-9 were significantly inferior to Examples 4-6 in terms of long-term stability, moisture content control, and disintegration time. Comparative Example 7 omitted polyethylene glycol 400, resulting in hygroscopic clumping and unsatisfactory uniformity, with a disintegration time extended to 23.5 minutes. Comparative Example 8 used only sucrose and PEG40, but after six months, the moisture content rose to 3.19%, and the disintegration time extended to 21.8 minutes, indicating that the sucrose-only system is highly hygroscopic and lacks stability. Comparative Example 9 used a combination of sucrose and hydroxypropyl-β-cyclodextrin (1:6). After six months, the moisture content was 2.22%, and the disintegration time was 20.9 minutes, inferior to Examples 4-6, likely due to delayed disintegration caused by starch gelatinization. By optimizing the excipient ratios, Examples 4-6 maintained a moisture content of ≤0.8% after six months of accelerated testing, and the disintegration time remained stable at approximately 15 minutes, demonstrating excellent stability and dissolution performance.

[0068] 3.2 [Content Determination] Determine according to the high performance liquid chromatography method (General Rule 0512).

[0069] Chromatographic conditions and system suitability test: octadecylsilane bonded silica gel was used as the filler; methanol-water-phosphorus (42:58:0.2) was used as the mobile phase; and the detection wavelength was 276 nm.

[0070] Preparation of reference solution: Take an appropriate amount of oroxylin B reference substance, accurately weigh it, and add 50% methanol to make a solution containing 0.10 mg per 1 mL.

[0071] Preparation of the test solution: Take 0.1 g of the capsules of Examples 4-6 and Comparative Examples 7-9, accurately weigh them, place them in a stoppered conical flask, accurately add 50 mL of methanol, weigh the weight, heat and reflux for 1 hour, cool, weigh again, make up the lost weight with methanol, shake well, filter, and take the filtrate to obtain the solution.

[0072] Determination method: Accurately aspirate 10 μL of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0073] Table 2 Oroxylin content in Examples 4-6 and Comparative Examples 7-9 As can be seen from the changes in orchidinoside content in Table 2, Examples 4-6 and Comparative Examples 7-9 exhibited significant differences after six months of accelerated testing. The orchidinoside content in Examples 4-6 remained stable, with only a slight decrease after six months, indicating that the formulation process and excipient ingredients effectively protected the active ingredient. However, Comparative Examples 7-9 showed significant degradation, suggesting that the optimization of different material systems may have accelerated the hydrolysis and oxidation of orchidinoside, leading to significant loss of the active ingredient during storage.

Claims

1. Use of a Chinese medicine composition in preparing a drug for treating osteosarcoma, characterized in that: The traditional Chinese medicine composition is prepared from Oroxylum indicum, Clematis chinensis, Scorpion, Hedgehog peel, Asarum, Cibotium barometz, Millennium Root, Panax notoginseng, Drynaria fortunei, Smilax glabra and Piper nigrum.

2. The use according to claim 1, characterized in that The components of the traditional Chinese medicine composition are: 30-70 parts of Oroxylum indicum, 10-30 parts of Clematis chinensis, 0.5-1.5 parts of Scorpio scorpion, 1-5 parts of Eryngium erinaceus, 1-3 parts of Asarum, 3-7 parts of Cibotium barometz, 3-7 parts of Millennium scutellaria, 8-12 parts of Panax notoginseng, 8-14 parts of Drynaria fortunei, 6-10 parts of Smilax glabra, and 5-9 parts of Piper mesenteriae.

3. The use according to claim 2, characterized in that The components of the traditional Chinese medicine composition are: 50 parts of Oroxylum indicum, 20 parts of Clematis chinensis, 1 part of Scorpio, 3 parts of Hedgehog peel, 2 parts of Asarum, 5 parts of Cibotium barometz, 5 parts of Millennium Root, 10 parts of Panax notoginseng, 11 parts of Drynaria fortunei, 8 parts of Smilax glabra, and 7 parts of Piper nigrum.

4. The use according to claim 1, wherein The components of the traditional Chinese medicine composition are: 30 parts of Oroxylum indicum, 10 parts of Clematis chinensis, 0.5 parts of Scorpio, 1 part of Eryngium erinaceus, 1 part of Asarum, 3 parts of Cibotium barometz, 3 parts of Millennium Root, 8 parts of Panax notoginseng, 8 parts of Drynaria fortunei, 6 parts of Smilax glabra, and 5 parts of Piper nigrum.

5. The use according to claim 1, characterized in that The components of the traditional Chinese medicine composition are: 70 parts of Oroxylum indicum, 30 parts of Clematis chinensis, 1.5 parts of Scorpio, 5 parts of Hedgehog peel, 3 parts of Asarum, 7 parts of Cibotium barometz, 7 parts of Millennium Root, 12 parts of Panax notoginseng, 14 parts of Drynaria fortunei, 10 parts of Smilax glabra, and 9 parts of Piper nigrum.

6. The use according to claim 2, characterized in that The Chinese medicine composition is an oral preparation; preferably, the oral preparation is a decoction, pills, tablets, mixture, capsule, granule or powder, more preferably a capsule or decoction.

7. The use according to claim 6, characterized in that The preparation method of the decoction comprises the following steps: (1) Pre-treatment step: Grind the whole scorpion and Panax notoginseng into fine powder respectively, so that the particle size is less than 100 μm; (2) Soak the Oroxylum indicum and the Herba Lycopodii in water for 2-5 hours to extract the volatile oil, add hydroxypropyl-β-cyclodextrin to prepare an inclusion compound, and keep the residue and aqueous solution for later use; (3) Water decoction step: wash the medicinal residue and aqueous solution of step (2), asarum, hedgehog skin, scalded dog spine and drynaria root, add water and decoct, wrap the asarum with gauze and add 10-15 times of the total weight of the medicinal materials, soak for 20-40 minutes, boil over high heat, then turn to low heat and decoct for 20-40 minutes, filter, and set aside the medicinal residue and liquid; add 8-10 times of the total weight of the medicinal materials to the medicinal residue, boil over high heat, then turn to low heat and decoct for 10-30 minutes, combine the two filtrates, and set aside the filter residue; (3) Alcohol extraction step: Clematidis, Millennium Jian, and Piper melongena were crushed into coarse powder, and then added with the whole scorpion from step (1) and the medicinal residue from step (3) and heated under 70% ethanol for 1-2 times, each time for 1.5-3.5 hours, and the alcohol extracts were combined and distilled under reduced pressure to obtain an alcohol extract; (4) Concentration and molding: Add the filtrate from step (2) to the alcohol extract from step (3), stir and dissolve continuously, and concentrate under reduced pressure to a relative density of 1.05-1.

11. Then add the fine powder of Panax notoginseng from step (1) and the inclusion compound from step (2), and stir to obtain a decoction.

8. The use according to claim 7, characterized in that The preparation method of the decoction is: (1) Pre-treatment step: Grind the whole scorpion and Panax notoginseng into fine powder respectively, so that the particle size is less than 100 μm; (2) Soak Oroxylum indicum and Herba Scutellariae in water for 3.5 hours to extract the volatile oil, add hydroxypropyl-β-cyclodextrin to prepare the inclusion compound, and keep the residue and aqueous solution for later use; (3) Water decoction step: wash the medicinal residue and aqueous solution of step (2), asarum, hedgehog skin, scalded dog spine and drynaria root, add water and decoct, wrap the asarum with gauze and add 13 times of the total weight of water to soak for 30 minutes, boil over high heat, then turn to low heat and decoct for 30 minutes, filter, and set aside the medicinal residue and liquid; add 9 times of the total weight of the medicinal materials to the medicinal residue, boil over high heat, then turn to low heat and decoct for 20 minutes, combine the two filtrates, and set aside the filter residue; (4) Alcohol extraction step: Clematidis, Qiannianjian and Kadzu are crushed into coarse powder, and then added with the whole scorpion from step (1) and the medicinal residue from step (3) and heated under 90% ethanol for reflux twice, each time for 3.5 hours, and the alcohol extracts are combined and distilled under reduced pressure to obtain an alcohol extract; (5) Concentration and molding: Add the filtrate from step (3) to the alcohol extract from step (4), stir and dissolve continuously, and concentrate under reduced pressure to a relative density of 1.

08. Then, add the fine powder of Panax notoginseng from step (1) and the inclusion compound from step (2), and stir to obtain a decoction.

9. The use according to claim 6, characterized in that The capsule preparation method comprises the following steps: S1: Extract preparation (1) Take the Chinese medicine composition described in any one of claims 2 to 5 and set aside; (2) Pre-treatment step: Grind the whole scorpion and Panax notoginseng into fine powder respectively, and control the particle size to be less than 100 μm; (3) Soak the Oroxylum indicum and the Herba Lycopodii in water for 2-5 hours, extract the volatile oil, add hydroxypropyl-β-cyclodextrin to prepare the inclusion compound, and set aside the dregs and aqueous solution. (4) Water decoction step: add the medicinal residue and aqueous solution of step (3), asarum, hedgehog skin, scalded dog spine, and drynaria root to water and decoct; wrap the asarum with gauze and add 13 times of the total weight of water to soak for 30 minutes; boil over high heat, then turn to low heat and decoct for 30 minutes; filter and set aside the medicinal residue and liquid for later use; add 9 times of the total weight of the medicinal materials to the medicinal residue and boil over high heat, then turn to low heat and decoct for 20 minutes; combine the two filtrates and set aside the filter residue for later use; (5) Alcohol extraction step: Clematidis, Qiannianjian and Kadzu are crushed into coarse powder, and then added with the whole scorpion of step (1) and the medicinal residue of step (4) to 90% or more of ethanol and heated under reflux twice, each time for 3.5 hours, and the alcohol extracts are combined and distilled under reduced pressure to obtain an alcohol extract; (6) Concentration and molding: add the filtrate of step (4) to the alcohol extract of step (5), add ethanol to make the alcohol content 50-70%, let it stand for 10-20 hours, filter, recover the ethanol under reduced pressure and concentrate to a relative density of 1.15-1.35 to obtain a thick paste, vacuum degree -0.08MPa--0.10MPa, drying temperature 85℃-95℃, crush under vacuum state and pass through 80 mesh sieve to obtain fine powder, set aside; S2: Capsule Preparation 1) Add the fine powder to polyethylene glycol 400, mix well to dissolve, and dry at 40-60°C; add a mixture of sucrose and hydroxypropyl-β-cyclodextrin in an amount of 63-77% by weight, wherein the weight ratio of sucrose to hydroxypropyl-β-cyclodextrin in the mixture is 4-8:1, add the Panax notoginseng fine powder of step (1) in S1 and the inclusion compound of step (3), add a binder to prepare a soft material, sieve through a 16-mesh sieve to granulate, boil and dry, and set aside; 2) Add an appropriate amount of magnesium stearate to the mixture in step (1) and mix evenly. Sieve through a 16-mesh sieve to obtain dry granules, which are then filled into capsule shells to prepare capsules.

10. The use according to claim 8, characterized in that The osteosarcoma is proximal femoral osteosarcoma.

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