Pharmaceutical composition containing sodium calcium edetate and dimercaptosuccinic acid as well as preparation method and application of pharmaceutical composition

Abstract

The invention discloses a pharmaceutical composition containing sodium calcium edetate and dimercaptosuccinic acid as well as a preparation method and application of the pharmaceutical composition, and relates to the technical field of biological medicines. The pharmaceutical composition comprises the following components: penthorum chinense pursh extract, tuber fleeceflower stem extract, anchusa, muskmelon pedicel, pomegranate root bark, dimercaptosuccinic acid and sodium calcium edetate. According to the invention, by carefully selecting each component and dosage and selecting proper traditional Chinese medicines and western medicines for mutual matching, the research finds that the pharmaceutical composition can effectively inhibit the growth of melanoma cells, is safe and free of side effects to human bodies after being used, and is suitable for large-area popularization and application.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to a pharmaceutical composition containing calcium sodium edetate and succimer, a preparation method and application thereof. Background Art

[0002] Melanoma is a malignant tumor formed by the malignant transformation of melanocytes. It can occur in the skin, extremities, mucous membranes, and other sites. However, due to its high metastatic potential and rapid progression, it is one of the leading causes of skin cancer-related mortality. Today, melanoma diagnosis and treatment have made significant progress both internationally and domestically. China has pioneered a unique approach to melanoma research that integrates local realities and promotes interdisciplinary innovation. However, it also faces numerous challenges, including the development of drug resistance, inadequate early diagnosis, treatment cost and accessibility, and the limitations of biomarkers.

[0003] If only traditional Chinese medicine is used to treat melanoma, although it can help improve or alleviate symptoms, it lacks the effect of directly inhibiting the proliferation or metastasis of melanoma cells and is slow to take effect. Melanoma is highly malignant and progresses rapidly. Relying solely on traditional Chinese medicine may delay the best time for treatment and lead to worsening of the disease. If only Western medicine is used to treat melanoma, chemotherapy drugs are often accompanied by serious adverse reactions such as bone marrow suppression and immune system damage, which may affect the patient's tolerance and quality of life. Some targeted drugs or chemotherapy regimens may produce drug resistance due to tumor cell mutations. The use of Western medicine alone is difficult to meet subsequent treatment needs. Therefore, there is an urgent need for a highly effective, safe and side-effect-free pharmaceutical composition that can inhibit the proliferation of melanoma. Summary of the Invention

[0004] In view of the above-mentioned defects of the prior art, the present invention proposes a pharmaceutical composition containing calcium sodium edetate and succimer, a preparation method and application thereof to solve the problems raised by the above-mentioned background technology.

[0005] In order to achieve the above object, the present invention provides the following technical solutions:

[0006] A pharmaceutical composition containing calcium sodium edetate and succimer, comprising the following ingredients by weight: 20-30 parts of ophiopogon scutellariae extract, 18-25 parts of smilax china extract, 10-20 parts of ophiopogon scutellariae, 8-16 parts of melon stalks, 5-10 parts of pomegranate root bark, 1-3 parts of succimer and 0.2-0.9 parts of calcium sodium edetate.

[0007] Preferably, the preparation method of the herb extract comprises the following steps: washing the underground part of the herb, drying it to a moisture content of 5 to 9 wt%, and then grinding it to 30 to 100 meshes to obtain herb powder; mixing the herb powder with an enzyme solution in a mass ratio of 1:(0.1 to 0.5), enzymolyzing it at a temperature of 50 to 55°C for 40 to 50 minutes, and then performing microwave treatment for 6 to 10 minutes, and drying it to obtain pretreated herb powder; adding the pretreated herb powder and tea polyphenols in a mass ratio of 1:(0.03 to 0.09) to an extraction kettle for supercritical CO2 extraction to obtain an extraction product; placing the extraction product in an environment of 2 to 5°C for a constant temperature of 100 to 150 minutes, then centrifuging to remove impurities, and evacuating it in a vacuum box at a temperature of 30 to 45°C for 20 to 50 minutes to obtain the herb extract.

[0008] Preferably, the enzyme solution is prepared by mixing cellulase, papain, and water in a mass ratio of (2-6):(3-8):(10-20).

[0009] Preferably, the power of the microwave is 300-600W, the pressure of the supercritical CO2 extraction is 30-40Mpa, the extraction temperature is 30-50°C, the extraction time is 50-180min, and the flow rate of the CO2 fluid is 10-25mL / min.

[0010] Preferably, the preparation method of the tuberculosis extract includes the following steps: crushing and sieving the tuberculosis to obtain coarse powder; adding 80% ethanol with a mass fraction of 80% ethanol that is 8 to 12 times the mass of the coarse powder to the coarse powder, extracting 2 to 3 times, each extraction lasting 30 to 60 minutes, combining the extracts, filtering, and obtaining a filtrate; placing the filtrate in a silica gel column, and eluting gradedly with a mixed solution of petroleum ether, chloroform, and ethyl acetate in a volume ratio of 1:1:1 as the mobile phase, and collecting the ethyl acetate elution phase, which is the tuberculosis extract.

[0011] Preferably, the pharmaceutical composition is in the form of an oral preparation or an external preparation.

[0012] Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

[0013] Preferably, the pharmaceutically acceptable excipients include one or more of excipients, binders, disintegrants, lubricants, coating agents, solvents, cosolvents, suspending agents, thickeners and surfactants.

[0014] Preferably, the preparation method of the pharmaceutical composition comprises the following steps:

[0015] (1) Weighing the extract of Herba Lycopodii, the extract of Herba Lycopodii, Herba Cynanchifoliae, the pedicle of Melon and the root bark of Pomegranate according to their weight to obtain a Chinese medicine mixture; adding water 10 to 17 times the total weight of the Chinese medicine mixture to the mixture, soaking for 100 to 120 minutes, then decocting for 30 to 50 minutes, filtering, repeating the decocting once for the residue after filtration, combining the filtrates, and evaporating and concentrating the filtrates to a relative density of 1.15 to 1.25 to obtain a concentrated solution;

[0016] (2) adding an equal volume of 95% ethanol to the concentrated solution obtained in step (1), mixing, refrigerating and standing at 2°C overnight, recovering the ethanol to obtain a medicinal solution, and concentrating the medicinal solution to a clear paste with a relative density of 1.35 to 1.38;

[0017] (3) Add succimer and sodium edetate to the clear paste prepared in step (2), mix well, and dry to obtain the target pharmaceutical composition.

[0018] Preferably, the pharmaceutical composition of the present application is used in the preparation of a drug for treating melanoma.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] (1) The present invention selects traditional Chinese medicine and Western medicine for treating melanoma and combines them to ensure that the prepared pharmaceutical composition can effectively inhibit the proliferation of melanoma. In addition, the selected ingredients are inexpensive and easily available. The preparation method of the pharmaceutical composition is simple to operate and does not require the assistance of large-scale equipment.

[0021] (2) The Herba Lycopodii extract of the present invention uses supercritical carbon dioxide fluid as a solvent for critical extraction, which has the characteristics of low-temperature extraction and inert protection. On the one hand, it effectively improves the extraction efficiency of the active ingredients of the Herba Lycopodii, and can extract the beneficial ingredients in the Herba Lycopodii to the maximum extent by extracting the Herba Lycopodii after processing. On the other hand, it prevents the oxidation and escape of "sensitive" substances, avoids the loss of activity of the beneficial ingredients, and can completely eliminate any organic solvents in the extraction process, thereby maintaining the naturalness of the extract.

[0022] (3) Dimercaptosuccinic acid and calcium sodium edetate are commonly used to treat heavy metal poisoning, and their application range is relatively narrow. This application proposes for the first time that dimercaptosuccinic acid and calcium sodium edetate can be used to treat melanoma, providing a new direction for the development of these two Western medicines and facilitating the subsequent development and application of dimercaptosuccinic acid and calcium sodium edetate. DETAILED DESCRIPTION

[0023] The technical solution of the present invention will be further described in detail below in conjunction with specific implementation methods.

[0024] Example 1

[0025] A pharmaceutical composition containing calcium sodium edetate and succimer, comprising the following ingredients by weight: 20 parts of an extract of Penthorum chinense Pursh (the whole herb of the Saxifragaceae plant Penthorum chinense Pursh), 18 parts of an extract of Fallopia multiflora (Thunb.) Harald. (the vine or vine with leaves of the Polygonaceae plant Fallopia multiflora (Thunb.) Harald.), 10 parts of a leaf of Rumex dentatus L. (the Polygonaceae plant), 8 parts of a fruit stalk of Cucumis melo L. (the Cucurbitaceae plant), 5 parts of a root bark of Punica granatum L. (the root and stem bark of the Punicaceae plant Punica granatum L., the Punicaceae family, the root and stem bark of which are used as medicine), 1 part of succimer, and calcium sodium edetate (C4H6O4S2). 10 H 12 CaN2Na2O8) 0.2 parts.

[0026] The preparation method of the herb extract comprises the following steps: washing the underground part of the herb, drying it to a moisture content of 5wt%, and then grinding it to 30 meshes to obtain herb powder; mixing the herb powder with an enzyme solution at a mass ratio of 1:0.1, performing enzymolysis at a temperature of 50°C for 40 minutes, then performing microwave treatment for 6 minutes, and drying to obtain pretreated herb powder; adding the pretreated herb powder and tea polyphenols at a mass ratio of 1:0.03 into an extraction kettle for supercritical CO2 extraction to obtain an extraction product; placing the extraction product in an environment of 2°C for a constant temperature of 100 minutes, then centrifuging to remove impurities, and evacuating the product in a vacuum box at a temperature of 30°C for 20 minutes to obtain the herb extract.

[0027] The enzyme solution is prepared by mixing cellulase, papain and water in a mass ratio of 2:3:10.

[0028] The power of the microwave is 300 W, the pressure of the supercritical CO2 extraction is 30 MPa, the extraction temperature is 30° C., the extraction time is 50 min, and the flow rate of the CO2 fluid is 10 mL / min.

[0029] The preparation method of the tuberculosis extract comprises the following steps: crushing and sieving the tuberculosis to obtain coarse powder; adding 80% ethanol with a mass fraction of 8 times the mass of the coarse powder to the coarse powder, extracting twice, each extraction lasting 30 minutes, combining the extracts, filtering, and obtaining a filtrate; placing the filtrate in a silica gel column, and graded eluting with a mixed solution of petroleum ether, chloroform, and ethyl acetate in a volume ratio of 1:1:1 as a mobile phase, and collecting the ethyl acetate elution phase to obtain the tuberculosis extract.

[0030] The dosage form of the pharmaceutical composition is any one of an oral preparation or an external preparation.

[0031] The pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0032] The pharmaceutically acceptable excipients include one or more of excipients, binders, disintegrants, lubricants, coating agents, solvents, cosolvents, suspending agents, thickeners and surfactants.

[0033] The preparation method of the pharmaceutical composition comprises the following steps:

[0034] (1) Weighing the extract of Herba Lycopodii, the extract of Herba Lycopodii, Herba Cynanchifoliae, the pedicle of Melon and the root bark of Pomegranate according to their weight to obtain a Chinese medicine mixture; adding water 10 times the total weight of the Chinese medicine mixture to the mixture and soaking for 100 minutes, then decocting for 30 minutes, filtering, repeating the decocting once for the residue after filtration, combining the filtrates, and evaporating and concentrating the filtrates to a relative density of 1.15 to obtain a concentrated solution;

[0035] (2) adding an equal volume of 95% ethanol to the concentrated solution obtained in step (1), mixing, refrigerating and standing at 2°C overnight, recovering the ethanol to obtain a medicinal solution, and concentrating the medicinal solution to a clear paste with a relative density of 1.35;

[0036] (3) Add succimer and sodium edetate to the clear paste prepared in step (2), mix well, and dry to obtain the target pharmaceutical composition.

[0037] Example 2

[0038] A pharmaceutical composition containing calcium sodium edetate and succimer, comprising the following ingredients in parts by weight: 30 parts of Herba Lycopodii Herba Extracts, 25 parts of Polygonum multiflorum Extracts, 20 parts of Herba Cynanchifoliae, 16 parts of Cucumis melo stalks, 10 parts of Pomegranate root bark, 3 parts of succimer and 0.9 parts of calcium sodium edetate.

[0039] The preparation method of the herb extract comprises the following steps: washing the underground part of the herb, drying it to a moisture content of 9wt%, and then grinding it to 100 meshes to obtain herb powder; mixing the herb powder with an enzyme solution at a mass ratio of 1:0.1, performing enzymolysis at a temperature of 55°C for 50 minutes, then performing microwave treatment for 10 minutes, and drying to obtain pretreated herb powder; adding the pretreated herb powder and tea polyphenols at a mass ratio of 1:0.09 into an extraction kettle for supercritical CO2 extraction to obtain an extraction product; placing the extraction product in an environment of 5°C at a constant temperature for 150 minutes, then centrifuging to remove impurities, and evacuating the product in a vacuum box at a temperature of 45°C for 50 minutes to obtain the herb extract.

[0040] The enzyme solution is prepared by mixing cellulase, papain and water in a mass ratio of 6:8:20.

[0041] The power of the microwave is 600 W, the pressure of the supercritical CO2 extraction is 40 MPa, the extraction temperature is 50° C., the extraction time is 180 min, and the flow rate of the CO2 fluid is 25 mL / min.

[0042] The preparation method of the tuberculosis extract comprises the following steps: crushing and sieving the tuberculosis to obtain coarse powder; adding 80% ethanol with a mass fraction of 12 times the mass of the coarse powder to the coarse powder, extracting three times, each extraction lasting 60 minutes, combining the extracts, filtering, and obtaining a filtrate; placing the filtrate in a silica gel column, and eluting by graded steps using a mixed solution of petroleum ether, chloroform, and ethyl acetate in a volume ratio of 1:1:1 as a mobile phase, and collecting the ethyl acetate elution phase, which is the tuberculosis extract.

[0043] The dosage form of the pharmaceutical composition is any one of an oral preparation or an external preparation.

[0044] The pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0045] The pharmaceutically acceptable excipients include one or more of excipients, binders, disintegrants, lubricants, coating agents, solvents, cosolvents, suspending agents, thickeners and surfactants.

[0046] The preparation method of the pharmaceutical composition comprises the following steps:

[0047] (1) Weighing the extract of Herba Lycopodii, the extract of Herba Lycopodii, Herba Cynanchifoliae, the pedicle of Melon and the root bark of Pomegranate according to their weight to obtain a Chinese medicine mixture; adding water 17 times the total weight of the Chinese medicine mixture to the mixture and soaking for 120 minutes, then decocting for 50 minutes, filtering, repeating the decocting once for the residue after filtration, combining the filtrates, and evaporating and concentrating the filtrates to a relative density of 1.25 to obtain a concentrate;

[0048] (2) adding an equal volume of 95% ethanol to the concentrated solution obtained in step (1), mixing, refrigerating and standing at 2°C overnight, recovering the ethanol to obtain a medicinal solution, and concentrating the medicinal solution to a clear paste with a relative density of 1.38;

[0049] (3) Add succimer and sodium edetate to the clear paste prepared in step (2), mix well, and dry to obtain the target pharmaceutical composition.

[0050] Example 3

[0051] A pharmaceutical composition containing calcium sodium edetate and succimer, comprising the following ingredients in parts by weight: 25 parts of Herba Lycopodii Herba Extracts, 20 parts of Polygonum multiflorum Extracts, 15 parts of Herba Cynanchifoliae, 10 parts of Cucumis melo stalks, 8 parts of Pomegranate root bark, 2 parts of succimer and 0.5 parts of calcium sodium edetate.

[0052] The preparation method of the herb extract comprises the following steps: washing the underground part of the herb, drying it to a moisture content of 8wt%, and then grinding it to 80 meshes to obtain herb powder; mixing the herb powder with an enzyme solution at a mass ratio of 1:0.4, performing enzymolysis at a temperature of 52°C for 45 minutes, then performing microwave treatment for 8 minutes, and drying to obtain pretreated herb powder; adding the pretreated herb powder and tea polyphenols at a mass ratio of 1:0.07 into an extraction kettle for supercritical CO2 extraction to obtain an extraction product; placing the extraction product in an environment of 3°C at a constant temperature for 120 minutes, then centrifuging to remove impurities, and evacuating the product in a vacuum box at a temperature of 40°C for 30 minutes to obtain the herb extract.

[0053] The enzyme solution is prepared by mixing cellulase, papain and water in a mass ratio of 4:7:15.

[0054] The power of the microwave is 500W, the pressure of the supercritical CO2 extraction is 35 MPa, the extraction temperature is 40°C, the extraction time is 100 min, and the flow rate of the CO2 fluid is 20 mL / min.

[0055] The preparation method of the tuberculosis extract comprises the following steps: crushing and sieving the tuberculosis to obtain coarse powder; adding 80% ethanol with a mass fraction of 10 times the mass of the coarse powder to the coarse powder, extracting three times, each extraction lasting 50 minutes, combining the extracts, filtering, and obtaining a filtrate; placing the filtrate in a silica gel column, and eluting the filtrate gradedly using a mixed solution of petroleum ether, chloroform, and ethyl acetate in a volume ratio of 1:1:1 as a mobile phase, and collecting the ethyl acetate elution phase to obtain the tuberculosis extract.

[0056] The dosage form of the pharmaceutical composition is any one of an oral preparation or an external preparation.

[0057] The pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0058] The pharmaceutically acceptable excipients include one or more of excipients, binders, disintegrants, lubricants, coating agents, solvents, cosolvents, suspending agents, thickeners and surfactants.

[0059] The preparation method of the pharmaceutical composition comprises the following steps:

[0060] (1) Weighing the extract of Herba Lycopodii, the extract of Herba Lycopodii, Herba Cynanchifoliae, the pedicle of Melon and the root bark of Pomegranate according to their weight to obtain a Chinese medicine mixture; adding water 15 times the total weight of the Chinese medicine mixture to the mixture and soaking for 110 minutes, then decocting for 40 minutes, filtering, repeating the decocting once for the residue after filtration, combining the filtrates, and evaporating and concentrating the filtrates to a relative density of 1.20 to obtain a concentrated solution;

[0061] (2) adding an equal volume of 95% ethanol to the concentrated solution obtained in step (1), mixing, refrigerating and standing at 2°C overnight, recovering the ethanol to obtain a medicinal solution, and concentrating the medicinal solution to a clear paste with a relative density of 1.37;

[0062] (3) Add succimer and sodium edetate to the clear paste prepared in step (2), mix well, and dry to obtain the target pharmaceutical composition.

[0063] Comparative Example 1

[0064] This comparative example differs from Example 3 in that the pharmaceutical composition does not contain the Herba Lycopodii extract, and the other ingredients and steps remain unchanged.

[0065] A pharmaceutical composition containing calcium sodium edetate and succimer, comprising the following ingredients in parts by weight: 20 parts of schizonepeta multiflora extract, 15 parts of ragweed, 10 parts of melon stalks, 8 parts of pomegranate root bark, 2 parts of succimer and 0.5 parts of calcium sodium edetate.

[0066] The preparation method of the tuberculosis extract comprises the following steps: crushing and sieving the tuberculosis to obtain coarse powder; adding 80% ethanol with a mass fraction of 10 times the mass of the coarse powder to the coarse powder, extracting three times, each extraction lasting 50 minutes, combining the extracts, filtering, and obtaining a filtrate; placing the filtrate in a silica gel column, and eluting the filtrate gradedly using a mixed solution of petroleum ether, chloroform, and ethyl acetate in a volume ratio of 1:1:1 as a mobile phase, and collecting the ethyl acetate elution phase to obtain the tuberculosis extract.

[0067] The dosage form of the pharmaceutical composition is any one of an oral preparation or an external preparation.

[0068] The pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0069] The pharmaceutically acceptable excipients include one or more of excipients, binders, disintegrants, lubricants, coating agents, solvents, cosolvents, suspending agents, thickeners and surfactants.

[0070] The preparation method of the pharmaceutical composition comprises the following steps:

[0071] (1) Weighing the extract of Polygonum multiflorum, Herba Cynanchifoliae, Melon pedicles, and Pomegranate root bark by weight to obtain a Chinese medicine mixture; adding 15 times the total weight of the Chinese medicine mixture in water to the mixture and soaking for 110 minutes, then decocting for 40 minutes, filtering, and repeatedly decocting the residue after filtration once, combining the filtrates, and evaporating and concentrating the filtrates to a relative density of 1.20 to obtain a concentrated solution;

[0072] (2) adding an equal volume of 95% ethanol to the concentrated solution obtained in step (1), mixing, refrigerating and standing at 2°C overnight, recovering the ethanol to obtain a medicinal solution, and concentrating the medicinal solution to a clear paste with a relative density of 1.37;

[0073] (3) Add succimer and sodium edetate to the clear paste prepared in step (2), mix well, and dry to obtain the target pharmaceutical composition.

[0074] Comparative Example 2

[0075] This comparative example differs from Example 3 in that the Herba Artemisiae Scopariae extract in the pharmaceutical composition is replaced with the Herba Artemisiae Scopariae extract, and the other ingredients and steps remain unchanged.

[0076] A pharmaceutical composition containing calcium sodium edetate and succimer, comprising the following ingredients in parts by weight: 25 parts of Artemisia capillaris extract, 20 parts of Polygonum multiflorum extract, 15 parts of Herba Cynanchifoliae, 10 parts of Cucumis melo stalks, 8 parts of Pomegranate root bark, 2 parts of succimer and 0.5 parts of calcium sodium edetate.

[0077] The preparation method of the artemisia capillaris extract comprises the following steps: washing the underground part of the artemisia capillaris, drying it to a moisture content of 8wt%, and then grinding it to 80 meshes to obtain artemisia capillaris powder; mixing the artemisia capillaris powder with an enzyme solution at a mass ratio of 1:0.4, performing enzymatic hydrolysis at a temperature of 52°C for 45 minutes, and then performing microwave treatment for 8 minutes to obtain pretreated artemisia capillaris powder; adding the pretreated artemisia capillaris powder and tea polyphenols at a mass ratio of 1:0.07 into an extraction kettle for supercritical CO2 extraction to obtain an extraction product; placing the extraction product in an environment of 3°C at a constant temperature for 120 minutes, then centrifuging to remove impurities, and evacuating the product in a vacuum box at a temperature of 40°C for 30 minutes to obtain the artemisia capillaris extract.

[0078] The enzyme solution is prepared by mixing cellulase, papain and water in a mass ratio of 4:7:15.

[0079] The power of the microwave is 500 W, the pressure of the supercritical CO2 extraction is 35 MPa, the extraction temperature is 40° C., the extraction time is 100 min, and the flow rate of the CO2 fluid is 20 mL / min.

[0080] The preparation method of the tuberculosis extract comprises the following steps: crushing and sieving the tuberculosis to obtain coarse powder; adding 80% ethanol with a mass fraction of 10 times the mass of the coarse powder to the coarse powder, extracting three times, each extraction lasting 50 minutes, combining the extracts, filtering, and obtaining a filtrate; placing the filtrate in a silica gel column, and eluting the filtrate gradedly using a mixed solution of petroleum ether, chloroform, and ethyl acetate in a volume ratio of 1:1:1 as a mobile phase, and collecting the ethyl acetate elution phase to obtain the tuberculosis extract.

[0081] The dosage form of the pharmaceutical composition is any one of an oral preparation or an external preparation.

[0082] The pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0083] The pharmaceutically acceptable excipients include one or more of excipients, binders, disintegrants, lubricants, coating agents, solvents, cosolvents, suspending agents, thickeners and surfactants.

[0084] The preparation method of the pharmaceutical composition comprises the following steps:

[0085] (1) Weighing the extract of Artemisia capillaris, the extract of Polygonum multiflorum, Herba Cynanchifoliae, Melon pedicles and Pomegranate root bark according to weight to obtain a Chinese medicine mixture; adding water 15 times the total weight of the Chinese medicine mixture to the mixture and soaking for 110 minutes, then decocting for 40 minutes, filtering, repeating the decocting once for the residue after filtration, combining the filtrates, and evaporating and concentrating the filtrates to a relative density of 1.20 to obtain a concentrated solution;

[0086] (2) adding an equal volume of 95% ethanol to the concentrated solution obtained in step (1), mixing, refrigerating and standing at 2°C overnight, recovering the ethanol to obtain a medicinal solution, and concentrating the medicinal solution to a clear paste with a relative density of 1.37;

[0087] (3) Add succimer and sodium edetate to the clear paste prepared in step (2), mix well, and dry to obtain the target pharmaceutical composition.

[0088] Comparative Example 3

[0089] This comparative example differs from Example 3 in that the preparation method of the Herba Lycopodii extract in the pharmaceutical composition is changed, while the other ingredients and steps remain unchanged.

[0090] A pharmaceutical composition containing calcium sodium edetate and succimer, comprising the following ingredients in parts by weight: 25 parts of Herba Lycopodii Herba Extracts, 20 parts of Polygonum multiflorum Extracts, 15 parts of Herba Cynanchifoliae, 10 parts of Cucumis melo stalks, 8 parts of Pomegranate root bark, 2 parts of succimer and 0.5 parts of calcium sodium edetate.

[0091] The preparation method of the Herba Lycopersicum var. truncatum extract comprises the following steps: washing the underground part of the Herba Lycopersicum var. truncatum, drying it to a moisture content of 8wt%, and then grinding it to 80 mesh to obtain Herba Lycopersicum var. truncatum powder; mixing the Herba Lycopersicum var. truncatum powder with an enzyme solution at a mass ratio of 1:0.4, performing enzymolysis at a temperature of 52°C for 45 minutes, and then performing microwave treatment for 8 minutes, and drying to obtain pretreated Herba Lycopersicum var. truncatum powder, namely the Herba Lycopersicum var. truncatum extract.

[0092] The enzyme solution is prepared by mixing cellulase, papain and water in a mass ratio of 4:7:15.

[0093] The power of the microwave is 500W.

[0094] The preparation method of the tuberculosis extract comprises the following steps: crushing and sieving the tuberculosis to obtain coarse powder; adding 80% ethanol with a mass fraction of 10 times the mass of the coarse powder to the coarse powder, extracting three times, each extraction lasting 50 minutes, combining the extracts, filtering, and obtaining a filtrate; placing the filtrate in a silica gel column, and graded eluting with a mixed solution of petroleum ether solution, chloroform solution, and ethyl acetate in a volume ratio of 1:1:1 as a mobile phase, and collecting the ethyl acetate elution phase to obtain the tuberculosis extract.

[0095] The dosage form of the pharmaceutical composition is any one of an oral preparation or an external preparation.

[0096] The pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0097] The pharmaceutically acceptable excipients include one or more of excipients, binders, disintegrants, lubricants, coating agents, solvents, cosolvents, suspending agents, thickeners and surfactants.

[0098] The preparation method of the pharmaceutical composition comprises the following steps:

[0099] (1) Weighing the extract of Herba Lycopodii, the extract of Herba Lycopodii, Herba Cynanchifoliae, the pedicle of Melon and the root bark of Pomegranate according to their weight to obtain a Chinese medicine mixture; adding water 15 times the total weight of the Chinese medicine mixture to the mixture and soaking for 110 minutes, then decocting for 40 minutes, filtering, repeating the decocting once for the residue after filtration, combining the filtrates, and evaporating and concentrating the filtrates to a relative density of 1.20 to obtain a concentrated solution;

[0100] (2) adding an equal volume of 95% ethanol to the concentrated solution obtained in step (1), mixing, refrigerating and standing at 2°C overnight, recovering the ethanol to obtain a medicinal solution, and concentrating the medicinal solution to a clear paste with a relative density of 1.37;

[0101] (3) Add succimer and sodium edetate to the clear paste prepared in step (2), mix well, and dry to obtain the target pharmaceutical composition.

[0102] Comparative Example 4

[0103] This comparative example differs from Example 3 in that the pharmaceutical composition does not contain the Polygonum multiflorum extract, and the other ingredients and steps remain unchanged.

[0104] A pharmaceutical composition containing calcium sodium edetate and succimer, comprising the following ingredients by weight: 25 parts of scutellaria baicalensis extract, 15 parts of scutellaria baicalensis, 10 parts of melon stalks, 8 parts of pomegranate root bark, 2 parts of succimer and 0.5 parts of calcium sodium edetate.

[0105] The preparation method of the herb extract comprises the following steps: washing the underground part of the herb, drying it to a moisture content of 8wt%, and then grinding it to 80 meshes to obtain herb powder; mixing the herb powder with an enzyme solution at a mass ratio of 1:0.4, performing enzymolysis at a temperature of 52°C for 45 minutes, then performing microwave treatment for 8 minutes, and drying to obtain pretreated herb powder; adding the pretreated herb powder and tea polyphenols at a mass ratio of 1:0.07 into an extraction kettle for supercritical CO2 extraction to obtain an extraction product; placing the extraction product in an environment of 3°C at a constant temperature for 120 minutes, then centrifuging to remove impurities, and evacuating the product in a vacuum box at a temperature of 40°C for 30 minutes to obtain the herb extract.

[0106] The enzyme solution is prepared by mixing cellulase, papain and water in a mass ratio of 4:7:15.

[0107] The power of the microwave is 500 W, the pressure of the supercritical CO2 extraction is 35 MPa, the extraction temperature is 40° C., the extraction time is 100 min, and the flow rate of the CO2 fluid is 20 mL / min.

[0108] The dosage form of the pharmaceutical composition is any one of an oral preparation or an external preparation.

[0109] The pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0110] The pharmaceutically acceptable excipients include one or more of excipients, binders, disintegrants, lubricants, coating agents, solvents, cosolvents, suspending agents, thickeners and surfactants.

[0111] The preparation method of the pharmaceutical composition comprises the following steps:

[0112] (1) Weighing the extract of Herba Cynanchifoliae, Herba Cynanchi, Melon Stem and Pomegranate Root Bark by weight to obtain a Chinese medicine mixture; adding 15 times the total weight of the Chinese medicine mixture in water to the mixture and soaking for 110 minutes, then decocting for 40 minutes, filtering, repeating the decocting once for the residue after filtration, combining the filtrates, and evaporating and concentrating the filtrates to a relative density of 1.20 to obtain a concentrate;

[0113] (2) adding an equal volume of 95% ethanol to the concentrated solution obtained in step (1), mixing, refrigerating and standing at 2°C overnight, recovering the ethanol to obtain a medicinal solution, and concentrating the medicinal solution to a clear paste with a relative density of 1.37;

[0114] (3) Add succimer and sodium edetate to the clear paste prepared in step (2), mix well, and dry to obtain the target pharmaceutical composition.

[0115] Comparative Example 5

[0116] This comparative example differs from Example 3 in that the preparation method of the Polygonum multiflorum extract in the pharmaceutical composition is changed, while the other ingredients and steps remain unchanged.

[0117] A pharmaceutical composition containing calcium sodium edetate and succimer, comprising the following ingredients in parts by weight: 25 parts of Herba Lycopodii Herba Extracts, 20 parts of Polygonum multiflorum Extracts, 15 parts of Herba Cynanchifoliae, 10 parts of Cucumis melo stalks, 8 parts of Pomegranate root bark, 2 parts of succimer and 0.5 parts of calcium sodium edetate.

[0118] The preparation method of the Herba Lycopersicum var. truncatum extract comprises the following steps: washing the underground part of the Herba Lycopersicum var. truncatum, drying it to a moisture content of 8wt%, and then grinding it to 80 mesh to obtain Herba Lycopersicum var. truncatum powder; mixing the Herba Lycopersicum var. truncatum powder with an enzyme solution at a mass ratio of 1:0.4, performing enzymolysis at a temperature of 52°C for 45 minutes, then performing microwave treatment for 8 minutes, and drying to obtain pretreated Herba Lycopersicum var. truncatum powder; adding the pretreated Herba Lycopersicum var. truncatum powder and tea polyphenols at a mass ratio of 1:0.07 into an extraction kettle for supercritical CO2 extraction to obtain an extraction product; placing the extraction product in an environment of 3°C at a constant temperature for 120 minutes, then centrifuging to remove impurities, and evacuating the product in a vacuum box at a temperature of 40°C for 30 minutes to obtain the Herba Lycopersicum var. truncatum extract;

[0119] The enzyme solution is prepared by mixing cellulase, papain and water in a mass ratio of 4:7:15.

[0120] The power of the microwave is 500W, the pressure of the supercritical CO2 extraction is 35 MPa, the extraction temperature is 40°C, the extraction time is 100 min, and the flow rate of the CO2 fluid is 20 mL / min.

[0121] The preparation method of the tuberculosis extract comprises the following steps: crushing and sieving the tuberculosis to obtain coarse powder; adding 80% ethanol with a mass fraction of 10 times the mass of the coarse powder to the coarse powder, extracting three times, each extraction lasting 50 minutes, combining the extracts, filtering to obtain a filtrate; and drying and concentrating the filtrate to obtain the tuberculosis extract.

[0122] The dosage form of the pharmaceutical composition is any one of an oral preparation or an external preparation.

[0123] The pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0124] The pharmaceutically acceptable excipients include one or more of excipients, binders, disintegrants, lubricants, coating agents, solvents, cosolvents, suspending agents, thickeners and surfactants.

[0125] The preparation method of the pharmaceutical composition comprises the following steps:

[0126] (1) Weighing the extract of Herba Lycopodii, the extract of Herba Lycopodii, Herba Cynanchifoliae, the pedicle of Melon and the root bark of Pomegranate according to their weight to obtain a Chinese medicine mixture; adding water 15 times the total weight of the Chinese medicine mixture to the mixture and soaking for 110 minutes, then decocting for 40 minutes, filtering, repeating the decocting once for the residue after filtration, combining the filtrates, and evaporating and concentrating the filtrates to a relative density of 1.20 to obtain a concentrated solution;

[0127] (2) adding an equal volume of 95% ethanol to the concentrated solution obtained in step (1), mixing, refrigerating and standing at 2°C overnight, recovering the ethanol to obtain a medicinal solution, and concentrating the medicinal solution to a clear paste with a relative density of 1.37;

[0128] (3) Add succimer and sodium edetate to the clear paste prepared in step (2), mix well, and dry to obtain the target pharmaceutical composition.

[0129] Comparative Example 6

[0130] This comparative example differs from Example 3 in that the pharmaceutical composition does not contain Herba Cynoglossi, and the other ingredients and steps remain unchanged.

[0131] A pharmaceutical composition containing calcium sodium edetate and succimer, comprising the following ingredients in parts by weight: 25 parts of Herba Lycopodii Herba Extracts, 20 parts of Polygonum multiflorum Extracts, 10 parts of Cucumis melo stalks, 8 parts of Pomegranate root barks, 2 parts of succimer and 0.5 parts of calcium sodium edetate.

[0132] The preparation method of the Herba Lycopersicum var. truncatum extract comprises the following steps: washing the underground part of the Herba Lycopersicum var. truncatum, drying it to a moisture content of 8wt%, and then grinding it to 80 mesh to obtain Herba Lycopersicum var. truncatum powder; mixing the Herba Lycopersicum var. truncatum powder with an enzyme solution at a mass ratio of 1:0.4, performing enzymolysis at a temperature of 52°C for 45 minutes, then performing microwave treatment for 8 minutes, and drying to obtain pretreated Herba Lycopersicum var. truncatum powder; adding the pretreated Herba Lycopersicum var. truncatum powder and tea polyphenols at a mass ratio of 1:0.07 into an extraction kettle for supercritical CO2 extraction to obtain an extraction product; placing the extraction product in an environment of 3°C at a constant temperature for 120 minutes, then centrifuging to remove impurities, and evacuating the product in a vacuum box at a temperature of 40°C for 30 minutes to obtain the Herba Lycopersicum var. truncatum extract;

[0133] The enzyme solution is prepared by mixing cellulase, papain and water in a mass ratio of 4:7:15.

[0134] The power of the microwave is 500 W, the pressure of the supercritical CO2 extraction is 35 MPa, the extraction temperature is 40° C., the extraction time is 100 min, and the flow rate of the CO2 fluid is 20 mL / min.

[0135] The preparation method of the tuberculosis extract comprises the following steps: crushing and sieving the tuberculosis to obtain coarse powder; adding 80% ethanol with a mass fraction of 10 times the mass of the coarse powder to the coarse powder, extracting three times, each extraction lasting 50 minutes, combining the extracts, filtering, and obtaining a filtrate; placing the filtrate in a silica gel column, and eluting the filtrate gradedly using a mixed solution of petroleum ether, chloroform, and ethyl acetate in a volume ratio of 1:1:1 as a mobile phase, and collecting the ethyl acetate elution phase to obtain the tuberculosis extract.

[0136] The dosage form of the pharmaceutical composition is any one of an oral preparation or an external preparation.

[0137] The pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0138] The pharmaceutically acceptable excipients include one or more of excipients, binders, disintegrants, lubricants, coating agents, solvents, cosolvents, suspending agents, thickeners and surfactants.

[0139] The preparation method of the pharmaceutical composition comprises the following steps:

[0140] (1) Weighing the extract of Herba Lycopodii, the extract of Caulis Smilacis Glabrae, the pedicle of Melon and the root bark of Pomegranate according to their weight to obtain a Chinese medicine mixture; adding water 15 times the total weight of the Chinese medicine mixture to the mixture and soaking for 110 minutes, then decocting for 40 minutes, filtering, repeating the decocting once for the residue after filtration, combining the filtrates, and evaporating and concentrating the filtrates to a relative density of 1.20 to obtain a concentrated solution;

[0141] (2) adding an equal volume of 95% ethanol to the concentrated solution obtained in step (1), mixing, refrigerating and standing at 2°C overnight, recovering the ethanol to obtain a medicinal solution, and concentrating the medicinal solution to a clear paste with a relative density of 1.37;

[0142] (3) Add succimer and sodium edetate to the clear paste prepared in step (2), mix well, and dry to obtain the target pharmaceutical composition.

[0143] Comparative Example 7

[0144] This comparative example differs from Example 3 in that the pharmaceutical composition does not contain muskmelon stalks and pomegranate root bark, and the other ingredients and steps remain unchanged.

[0145] A pharmaceutical composition containing calcium sodium edetate and succimer, comprising the following ingredients in parts by weight: 25 parts of Herba Lycopodii Herba Extracts, 20 parts of Polygonum multiflorum Extracts, 15 parts of Herba Cynanchifoliae, 2 parts of succimer and 0.5 parts of calcium sodium edetate.

[0146] The preparation method of the Herba Lycopersicum var. truncatum extract comprises the following steps: washing the underground part of the Herba Lycopersicum var. truncatum, drying it to a moisture content of 8wt%, and then grinding it to 80 mesh to obtain Herba Lycopersicum var. truncatum powder; mixing the Herba Lycopersicum var. truncatum powder with an enzyme solution at a mass ratio of 1:0.4, performing enzymolysis at a temperature of 52°C for 45 minutes, then performing microwave treatment for 8 minutes, and drying to obtain pretreated Herba Lycopersicum var. truncatum powder; adding the pretreated Herba Lycopersicum var. truncatum powder and tea polyphenols at a mass ratio of 1:0.07 into an extraction kettle for supercritical CO2 extraction to obtain an extraction product; placing the extraction product in an environment of 3°C at a constant temperature for 120 minutes, then centrifuging to remove impurities, and evacuating the product in a vacuum box at a temperature of 40°C for 30 minutes to obtain the Herba Lycopersicum var. truncatum extract;

[0147] The enzyme solution is prepared by mixing cellulase, papain and water in a mass ratio of 4:7:15.

[0148] The power of the microwave is 500 W, the pressure of the supercritical CO2 extraction is 35 MPa, the extraction temperature is 40° C., the extraction time is 100 min, and the flow rate of the CO2 fluid is 20 mL / min.

[0149] The preparation method of the tuberculosis extract comprises the following steps: crushing and sieving the tuberculosis to obtain coarse powder; adding 80% ethanol with a mass fraction of 10 times the mass of the coarse powder to the coarse powder, extracting three times, each extraction lasting 50 minutes, combining the extracts, filtering, and obtaining a filtrate; placing the filtrate in a silica gel column, and eluting the filtrate gradedly using a mixed solution of petroleum ether, chloroform, and ethyl acetate in a volume ratio of 1:1:1 as a mobile phase, and collecting the ethyl acetate elution phase to obtain the tuberculosis extract.

[0150] The dosage form of the pharmaceutical composition is any one of an oral preparation or an external preparation.

[0151] The pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0152] The pharmaceutically acceptable excipients include one or more of excipients, binders, disintegrants, lubricants, coating agents, solvents, cosolvents, suspending agents, thickeners and surfactants.

[0153] The preparation method of the pharmaceutical composition comprises the following steps:

[0154] (1) Weighing the extract of Herba Lysimachiae, the extract of Herba Lysimachiae and Herba Cynanchii in parts by weight to obtain a Chinese medicine mixture; adding water 15 times the total weight of the Chinese medicine mixture to the mixture and soaking for 110 minutes, then decocting for 40 minutes, filtering, repeating the decocting once for the residue after filtration, combining the filtrates, and evaporating and concentrating the filtrates to a relative density of 1.20 to obtain a concentrated solution;

[0155] (2) adding an equal volume of 95% ethanol to the concentrated solution obtained in step (1), mixing, refrigerating and standing at 2°C overnight, recovering the ethanol to obtain a medicinal solution, and concentrating the medicinal solution to a clear paste with a relative density of 1.37;

[0156] (3) Add succimer and sodium edetate to the clear paste prepared in step (2), mix well, and dry to obtain the target pharmaceutical composition.

[0157] Comparative Example 8

[0158] This comparative example differs from Example 3 in that the pharmaceutical composition does not contain succimer, and the other ingredients and steps remain unchanged.

[0159] A pharmaceutical composition containing calcium sodium edetate comprises the following ingredients in parts by weight: 25 parts of radix scutellariae extract, 20 parts of scutellariae extract, 15 parts of scutellariae scutellariae, 10 parts of melon stalks, 8 parts of pomegranate root bark and 0.5 parts of calcium sodium edetate.

[0160] The preparation method of the Herba Lycopersicum var. truncatum extract comprises the following steps: washing the underground part of the Herba Lycopersicum var. truncatum, drying it to a moisture content of 8wt%, and then grinding it to 80 mesh to obtain Herba Lycopersicum var. truncatum powder; mixing the Herba Lycopersicum var. truncatum powder with an enzyme solution at a mass ratio of 1:0.4, performing enzymolysis at a temperature of 52°C for 45 minutes, then performing microwave treatment for 8 minutes, and drying to obtain pretreated Herba Lycopersicum var. truncatum powder; adding the pretreated Herba Lycopersicum var. truncatum powder and tea polyphenols at a mass ratio of 1:0.07 into an extraction kettle for supercritical CO2 extraction to obtain an extraction product; placing the extraction product in an environment of 3°C at a constant temperature for 120 minutes, then centrifuging to remove impurities, and evacuating the product in a vacuum box at a temperature of 40°C for 30 minutes to obtain the Herba Lycopersicum var. truncatum extract;

[0161] The enzyme solution is prepared by mixing cellulase, papain and water in a mass ratio of 4:7:15.

[0162] The power of the microwave is 500 W, the pressure of the supercritical CO2 extraction is 35 MPa, the extraction temperature is 40° C., the extraction time is 100 min, and the flow rate of the CO2 fluid is 20 mL / min.

[0163] The preparation method of the tuberculosis extract comprises the following steps: crushing and sieving the tuberculosis to obtain coarse powder; adding 80% ethanol with a mass fraction of 10 times the mass of the coarse powder to the coarse powder, extracting three times, each extraction lasting 50 minutes, combining the extracts, filtering, and obtaining a filtrate; placing the filtrate in a silica gel column, and eluting the filtrate gradedly using a mixed solution of petroleum ether, chloroform, and ethyl acetate in a volume ratio of 1:1:1 as a mobile phase, and collecting the ethyl acetate elution phase to obtain the tuberculosis extract.

[0164] The dosage form of the pharmaceutical composition is any one of an oral preparation or an external preparation.

[0165] The pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0166] The pharmaceutically acceptable excipients include one or more of excipients, binders, disintegrants, lubricants, coating agents, solvents, cosolvents, suspending agents, thickeners and surfactants.

[0167] The preparation method of the pharmaceutical composition comprises the following steps:

[0168] (1) Weighing the extract of Herba Lycopodii, the extract of Herba Lycopodii, Herba Cynanchifoliae, the pedicle of Melon and the root bark of Pomegranate according to their weight to obtain a Chinese medicine mixture; adding water 15 times the total weight of the Chinese medicine mixture to the mixture and soaking for 110 minutes, then decocting for 40 minutes, filtering, repeating the decocting once for the residue after filtration, combining the filtrates, and evaporating and concentrating the filtrates to a relative density of 1.20 to obtain a concentrated solution;

[0169] (2) adding an equal volume of 95% ethanol to the concentrated solution obtained in step (1), mixing, refrigerating and standing at 2°C overnight, recovering the ethanol to obtain a medicinal solution, and concentrating the medicinal solution to a clear paste with a relative density of 1.37;

[0170] (3) Adding calcium sodium edetate to the clear paste prepared in step (2), mixing, and drying to obtain the target pharmaceutical composition.

[0171] Comparative Example 9

[0172] This comparative example differs from Example 3 in that the pharmaceutical composition does not contain edetate calcium sodium, and the other ingredients and steps remain unchanged.

[0173] A pharmaceutical composition containing dithiothreitol comprises the following ingredients in parts by weight: 25 parts of Herba Lycopodii Herba Extracts, 20 parts of Polygonum multiflorum Extracts, 15 parts of Herba Cynanchifoliae, 10 parts of Cucumis melo pedicles, 8 parts of Pomegranate root barks and 2 parts of dithiothreitol.

[0174] The preparation method of the Herba Lycopersicum var. truncatum extract comprises the following steps: washing the underground part of the Herba Lycopersicum var. truncatum, drying it to a moisture content of 8wt%, and then grinding it to 80 mesh to obtain Herba Lycopersicum var. truncatum powder; mixing the Herba Lycopersicum var. truncatum powder with an enzyme solution at a mass ratio of 1:0.4, performing enzymolysis at a temperature of 52°C for 45 minutes, then performing microwave treatment for 8 minutes, and drying to obtain pretreated Herba Lycopersicum var. truncatum powder; adding the pretreated Herba Lycopersicum var. truncatum powder and tea polyphenols at a mass ratio of 1:0.07 into an extraction kettle for supercritical CO2 extraction to obtain an extraction product; placing the extraction product in an environment of 3°C at a constant temperature for 120 minutes, then centrifuging to remove impurities, and evacuating the product in a vacuum box at a temperature of 40°C for 30 minutes to obtain the Herba Lycopersicum var. truncatum extract;

[0175] The enzyme solution is prepared by mixing cellulase, papain and water in a mass ratio of 4:7:15.

[0176] The power of the microwave is 500 W, the pressure of the supercritical CO2 extraction is 35 MPa, the extraction temperature is 40° C., the extraction time is 100 min, and the flow rate of the CO2 fluid is 20 mL / min.

[0177] The preparation method of the tuberculosis extract comprises the following steps: crushing and sieving the tuberculosis to obtain coarse powder; adding 80% ethanol with a mass fraction of 10 times the mass of the coarse powder to the coarse powder, extracting three times, each extraction lasting 50 minutes, combining the extracts, filtering, and obtaining a filtrate; placing the filtrate in a silica gel column, and eluting the filtrate gradedly using a mixed solution of petroleum ether, chloroform, and ethyl acetate in a volume ratio of 1:1:1 as a mobile phase, and collecting the ethyl acetate elution phase to obtain the tuberculosis extract.

[0178] The dosage form of the pharmaceutical composition is any one of an oral preparation or an external preparation.

[0179] The pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0180] The pharmaceutically acceptable excipients include one or more of excipients, binders, disintegrants, lubricants, coating agents, solvents, cosolvents, suspending agents, thickeners and surfactants.

[0181] The preparation method of the pharmaceutical composition comprises the following steps:

[0182] (1) Weighing the extract of Herba Lycopodii, the extract of Herba Lycopodii, Herba Cynanchifoliae, the pedicle of Melon and the root bark of Pomegranate according to their weight to obtain a Chinese medicine mixture; adding water 15 times the total weight of the Chinese medicine mixture to the mixture and soaking for 110 minutes, then decocting for 40 minutes, filtering, repeating the decocting once for the residue after filtration, combining the filtrates, and evaporating and concentrating the filtrates to a relative density of 1.20 to obtain a concentrated solution;

[0183] (2) adding an equal volume of 95% ethanol to the concentrated solution obtained in step (1), mixing, refrigerating and standing at 2°C overnight, recovering the ethanol to obtain a medicinal solution, and concentrating the medicinal solution to a clear paste with a relative density of 1.37;

[0184] (3) Add succimer to the clear paste prepared in step (2), mix well, and dry to obtain the target pharmaceutical composition.

[0185] Experimental Example 1 In vitro experiment on inhibiting proliferation of human melanoma SK-MEL-5 cells

[0186] 1.1 Experimental Materials

[0187] The pharmaceutical compositions prepared in Examples 1 to 3 and Comparative Examples 1 to 9; human melanoma SK-MEL-5 cells were purchased from Shanghai Fuyu Biotechnology Co., Ltd. and cultured in DME M (Dulbecco's modified eagle medium) containing 10% FBS (fetal bovine serum) by volume at a constant temperature of 37°C and an ambient CO2 concentration of 5% for later use; Vemurafenib tablets were purchased from Zhengzhou Baorentang Pharmaceutical Technology Co., Ltd.

[0188] 1.2 Experimental methods

[0189] Melanoma SK MEL 5 cells were cultured at 5×10 3 Each well was inoculated in a 96-well plate and cultured (10% FBS / DMEM, ambient temperature 37°C, CO2 concentration 5%), divided into experimental group, positive control group and blank control group, with 5 replicates per group. After 14-16 hours of culture, pharmaceutical composition (12 mmol / L) was added to the culture medium of the experimental group (pharmaceutical composition prepared by Examples 1-3 and Comparative Examples 1-9), and vemurafenib was added to the positive control group so that the final concentration of the pharmaceutical composition in the experimental group culture medium and the final concentration of vemurafenib in the positive control group were 12 μmol / L. The blank control group was only added with culture medium without the addition of pharmaceutical composition. CCK8 was added to both the experimental group and the control group at 48 hours and incubated in an incubator for 2 hours, and then the absorbance (OD value) of each well at a wavelength of 450 nm was detected by microplate reader.

[0190] 1.3 Experimental Results

[0191] The inhibitory effect of the pharmaceutical composition prepared in the present application on the proliferation of melanoma cells was detected by CCK8, and the results are shown in Table 1:

[0192] Table 1 Proliferation results of human melanoma SK-MEL-5 cells

[0193]

[0194]

[0195] As can be seen from Table 1, the pharmaceutical composition prepared in the present application has an inhibitory effect on the proliferation of human melanoma SK-MEL-5 cells. The survival rate of human melanoma SK-MEL-5 cells in Examples 1 to 3 is only 9.5% to 12.4%, the survival rate of human melanoma SK-MEL-5 cells in the positive control group is 21.7%, and the survival rate of human melanoma SK-MEL-5 cells in Comparative Examples 1 to 9 is 33.4% to 60.3%. The blank control group has no effect on the proliferation of human melanoma SK-MEL-5 cells. It can be seen that the pharmaceutical composition prepared in the present application has a better inhibitory effect on human melanoma SK-MEL-5 cells than the positive control group. Comparison of Example 3 with Comparative Examples 1 to 5 shows that the components of the Herba Lycopodii Extract and the Herba Polygoni Multiflori Extract and their preparation methods have a significant effect on the efficacy of the pharmaceutical composition. If the Herba Lycopodii Extract and the Herba Polygoni Multiflori Extract are not added, the efficacy of the pharmaceutical composition will decrease. Even if the Herba Lycopodii is replaced with Artemisia Capillaris, which has similar pharmacological properties, it cannot replace the role of the Herba Lycopodii in the pharmaceutical composition. If the Herba Lycopodii Extract and the Herba Polygoni Multiflori Extract are not subjected to supercritical carbon dioxide extraction and silica gel column elution respectively during the preparation process, the active ingredients in the extract will be lost or the beneficial ingredients will not be extracted. Comparison of Example 3 with Comparative Examples 6 to 9 shows that the various components in the pharmaceutical composition of the present application are carefully selected and added by the applicant. If a certain component is missing, the inhibitory effect on the proliferation of melanoma cells can be reduced.

[0196] Experimental Example 2 Effects on Tumor Growth in B16F10 Tumor-Bearing C57BL / 6 Mice

[0197] 2.1 Experimental Materials

[0198] The pharmaceutical compositions prepared in Example 3 and Comparative Examples 1 to 9; human melanoma SK-MEL-5 cells were purchased from Shanghai Fuyu Biotechnology Co., Ltd. and cultured in DMEM (Dulbecco's modified eagle medium) containing 10% FBS (fetal bovine serum) by volume, at a constant temperature of 37°C and an ambient CO2 concentration of 5% for later use; Vemurafenib tablets were purchased from Zhengzhou Baorentang Pharmaceutical Technology Co., Ltd.

[0199] 2.2 Experimental methods

[0200] Ten C57BL / 6 mice were shaved on their backs and subcutaneously inoculated with mouse melanoma B16F10 cells. Once palpable tumors were achieved, the mice were randomly divided into 12 groups of 10 mice each. The experimental group was gavaged with the pharmaceutical composition prepared in Example 3 and Comparative Examples 1-9 at a dose of 15 mg / kg, while the positive control group was gavaged with vemurafenib tablets at a dose of 15 mg / kg. The blank control group was gavaged with distilled water. Ten days after gavage, the mice's body weight and tumor volume were measured until the study endpoint. Tumor volume inhibition (TGI) = (1 - tumor weight in the experimental group / tumor weight in the blank control group) × 100%.

[0201] 2.3 Experimental Results

[0202] Table 2 Tumor growth

[0203]

[0204] As shown in Table 2, the pharmaceutical compositions prepared herein can effectively inhibit melanoma tumor volume and prevent tumor enlargement. Furthermore, experiments revealed that no mice died after taking the pharmaceutical compositions prepared in Example 3 and Comparative Examples 1-9, and the weights of mice taking the pharmaceutical compositions were similar to those in the blank control group, indicating that the pharmaceutical compositions did not affect mouse growth and had no significant side effects.

[0205] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A pharmaceutical composition containing calcium sodium edetate and succimer, characterized in that: The invention comprises the following ingredients in parts by weight: 20-30 parts of Herba Lycopodii Herba Extract, 18-25 parts of Polygonum multiflorum Extract, 10-20 parts of Herba Cynoglossi, 8-16 parts of Cucumis melo pedicles, 5-10 parts of Pomegranate root bark, 1-3 parts of succimer dithiocarpa and 0.2-0.9 parts of sodium calcium edetate.

2. The pharmaceutical composition according to claim 1, characterized in that The preparation method of the herb extract comprises the following steps: washing the underground part of the herb, drying it to a moisture content of 5-9wt%, and then grinding it to 30-100 meshes to obtain herb powder; mixing the herb powder with an enzyme solution at a mass ratio of 1:(0.1-0.5), performing enzymatic hydrolysis at a temperature of 50-55°C for 40-50 minutes, then performing microwave treatment for 6-10 minutes, and drying to obtain pretreated herb powder; adding the pretreated herb powder and tea polyphenols at a mass ratio of 1:(0.03-0.09) into an extraction kettle for supercritical CO2 extraction to obtain an extraction product; placing the extraction product in an environment of 2-5°C for a constant temperature of 100-150 minutes, then centrifuging to remove impurities, and evacuating the mixture in a vacuum box at a temperature of 30-45°C for 20-50 minutes to obtain the herb extract.

3. The pharmaceutical composition according to claim 2, characterized in that The enzyme solution is prepared by mixing cellulase, papain and water in a mass ratio of (2-6):(3-8):(10-20).

4. The pharmaceutical composition according to claim 2, characterized in that The power of the microwave is 300-600W, the pressure of the supercritical CO2 extraction is 30-40Mpa, the extraction temperature is 30-50°C, the extraction time is 50-180min, and the flow rate of the CO2 fluid is 10-25mL / min.

5. The pharmaceutical composition according to claim 1, characterized in that The preparation method of the tuberculosis extract comprises the following steps: crushing and sieving the tuberculosis to obtain coarse powder; adding 80% ethanol with a mass fraction of 80% ethanol, which is 8 to 12 times the mass of the coarse powder, to the coarse powder, extracting for 2 to 3 times, each extraction lasting 30 to 60 minutes, combining the extracts, filtering, and obtaining a filtrate; placing the filtrate in a silica gel column, and eluting the filtrate gradedly using a mixed solution of petroleum ether, chloroform, and ethyl acetate in a volume ratio of 1:1:1 as a mobile phase, and collecting the ethyl acetate elution phase, which is the tuberculosis extract.

6. The pharmaceutical composition according to claim 1, characterized in that The dosage form of the pharmaceutical composition is any one of an oral preparation or an external preparation.

7. The pharmaceutical composition according to claim 1, characterized in that The pharmaceutical composition further includes pharmaceutically acceptable excipients.

8. The pharmaceutical composition according to claim 7, characterized in that The pharmaceutically acceptable excipients include one or more of excipients, binders, disintegrants, lubricants, coating agents, solvents, cosolvents, suspending agents, thickeners and surfactants.

9. The method for preparing the pharmaceutical composition according to claim 1, wherein The following steps are involved: (1) Weighing the extract of Herba Lycopodii, the extract of Herba Lycopodii, Herba Cynanchifoliae, the pedicle of Melon and the root bark of Pomegranate according to their weight to obtain a Chinese medicine mixture; adding water 10 to 17 times the total weight of the Chinese medicine mixture to the mixture, soaking for 100 to 120 minutes, then decocting for 30 to 50 minutes, filtering, repeating the decocting once for the residue after filtration, combining the filtrates, and evaporating and concentrating the filtrates to a relative density of 1.15 to 1.25 to obtain a concentrated solution; (2) adding an equal volume of 95% ethanol to the concentrated solution obtained in step (1), mixing, refrigerating and standing at 2°C overnight, recovering the ethanol to obtain a medicinal solution, and concentrating the medicinal solution to a clear paste with a relative density of 1.35 to 1.38; (3) Add succimer and sodium edetate to the clear paste prepared in step (2), mix well, and dry to obtain the target pharmaceutical composition.

10. Use of the pharmaceutical composition according to any one of claims 1 to 8 or the pharmaceutical composition prepared by the preparation method according to claim 9 in preparing a drug for treating melanoma.

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