A traditional Chinese medicine composition of Jinwugongtong and a preparation method and application thereof

Abstract

The present application relates to a kind of traditional Chinese medicine composition of Jinyu Gutong and preparation method and application, the composition and preparation are made from active ingredients of dog spine, epimedium, earth party member etc..By reasonable lord vassal scribe composition prescription, mainly is to exert the efficacy of active ingredients of dog spine, fuzhriteng, epimedium, earth cow knee, wilson wonder, radix puerariae, expelling wind and removing dampness, strengthening sinew and activating collateral, preventing and treating osteoporosis, again combining the efficacy of active ingredients of papaya and turmeric, and the efficacy of wucaishesheng, such as anti-inflammatory and bactericidal, expelling wind and dredging collaterals and antispasmodic, and the efficacy of earth party member active ingredient, spleen and qi, plus the dosage of specific proportion of drug, make the whole composition achieve synergistic effect.Compared with prior art, the present application carries out secondary development research on the composition of "Jinyu Gutong Capsule", improves the effect of expelling wind and removing dampness, promoting blood circulation and removing obstruction in the collaterals, maximizes the drug efficacy on lumbar and leg soreness, limb numbness, treatment of osteoporosis and other symptoms caused by wind-cold-dampness arthralgia, and does not produce drug dependence, non-toxic side effects.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine compositions, specifically to a Jinwu Gutong traditional Chinese medicine composition, its preparation method, and its application. Background Technology

[0002] "Jinwu Gutong Capsules" originate from a classic Miao medicine formula and are a core Miao medicine product of Guizhou Shengshi Longfang Pharmaceutical Co., Ltd. It is made from a formula of Cibotium barometz, Epimedium, Codonopsis pilosula, Psoralea corylifolia, Chaenomeles speciosa, Curcuma longa, Pueraria lobata, Achyranthes bidentata, Clematis chinensis, and Zaocys dhumnades. It nourishes the liver and kidneys, dispels wind and dampness, and invigorates blood circulation (Miao medicine: Wei Xiang Yang Diu Xiang, Yang An Dang Meng, Jiang Shi Feng, Gao Ji Tiao Ga Bian Meng). It is used for symptoms such as lower back and leg pain and limb numbness caused by liver and kidney deficiency, wind-cold-dampness syndrome, osteoporosis, and bone hyperplasia. It is included in the "National Compendium of Standard Chinese Patent Medicines: Brain System, Meridians, and Limbs" and the "National Medical Insurance" (2022 edition).

[0003] In 2005, the inventors conducted research on Jinwu Gutong tablets and applied for a Chinese invention patent for Jinwu Gutong tablets and its preparation method (patent number CN200510003185.2). This invention patent mainly studies Jinwu Gutong tablets and its preparation process, focusing on the proportion and dosage of each Chinese medicinal material in the formula, the separate drying of different medicinal materials, multiple decoctions, and the final concentration and mixing. The preparation drug of this invention has achieved the expected therapeutic effect and has also enriched the variety of Jinwu Gutong compositions that only have a single dosage form of capsules, making it convenient for different patients to choose to take it according to their preference for dosage form. Because this invention did not study the active ingredients of the medicinal materials in the formulation, and due to historical reasons, the preparation method of Jinwu Gutong Capsules did not specify the exact amount of solvent added or other specific process parameters, and the quality standards did not require the amount of active ingredients in the medicinal materials, the inventors, after years of research and practice, proposed a new approach to study the active ingredients of the Jinwu Gutong composition. The main advancements of this invention compared to the inventors' original products are: first, it clarifies the active ingredients of each medicinal material in the Jinwu Gutong formulation that exert its medicinal effect; second, it accurately determines the amount of each component in the composition; and third, it reduces the potential side effects caused by the complexity of the raw materials of Chinese medicinal materials.

[0004] In existing technologies, such as Chinese invention patent CN105663901A, a traditional Chinese medicine preparation for treating degenerative osteoarthritis is disclosed. Its raw materials include 29 kinds of herbs such as white peony root, angelica root, chuanxiong rhizome, salvia root, processed pangolin scales, notoginseng, raw astragalus root, raw licorice root, coix seed, cistanche, cibotium barometz, achyranthes root, kudzu root, trichosanthes root, raw eucommia bark, drynaria rhizome, morinda root, epimedium, mulberry mistletoe, agkistrodon halys, processed aconite root, corydalis rhizome, raw rhubarb, gentian root, fangji root, raw rehmannia root, asarum root, angelica pubescens root, and chicken blood vine. This formula uses as many as 29 kinds of Chinese medicinal materials, making the formula too complicated and the effective components unclear.

[0005] At present, the active ingredient research of traditional Chinese medicine composition and the low content of active ingredient, poor bioavailability of traditional Chinese medicine composition are common, and the research on other related drugs and their extracts has not been carried out in depth. In recent years, the extensive research on the chemical composition, pharmacological action and application of traditional Chinese medicine at home and abroad makes the active ingredient of traditional Chinese medicine an important way for new drug research and development. As we all know, traditional Chinese medicine believes that "medicine is three parts of poison", if not reasonably matched and processed, it will produce side effects in treatment, and the effective components obtained from the extraction process of traditional Chinese medicine raw materials are not clear, so the dosage cannot be accurately used, the drug dosage is large and the effect is slow. It can be seen that it is meaningful to upgrade and improve the existing products on the market, to clarify and fully utilize the effective active ingredients of components, and it is an important research direction for us to further optimize the production process and safely and effectively control the quality of drugs.

[0006] The inventor changes the traditional use of Chinese herbal medicine raw materials on the basis of the research of "UPLC-Q-Exactive-MS / MS combined with network pharmacology to explore the active ingredients and mechanism of action of JINU GUTONG capsule in the treatment of osteoporosis", and carries out effective active ingredient research on JINU GUTONG composition, selects the core component of the effective active ingredient of Chinese herbal medicine for traditional Chinese medicine composition protection, clarifies the effective components of the prescription, and accurately uses the amount, so as to overcome the defects of the existing products that most of the components of the prescription are more, the main and secondary effects are not obvious, or the product has multiple effects, but the actual effect is not ideal. Therefore, the present study carries out secondary development research on the prescription of "JINU GUTONG capsule", and scientifically selects the active ingredients of dog spine, icariin and icariin, the active ingredients of radix ginseng, the active ingredients of isopsoralen, the active ingredients of papaya polyphenol, the active ingredients of curcumin, the active ingredients of pueraria, the active ingredients of dehydroandrostanol, the active ingredients of radix clematidis and ophiophagus, clarifies the effective components of the prescription, accurately uses the amount of prescription, and promotes the development of modernization of traditional Chinese medicine. SUMMARY

[0007] In order to solve the above technical problems existing in the prior art, the present application provides a JINU GUTONG traditional Chinese medicine composition and its preparation method and application, which selects the effective components of traditional Chinese medicine, clarifies the effective component dosage and effect, and has a positive effect on the modernization development of traditional Chinese medicine of "JINU GUTONG capsule".

[0008] Specifically, the present application is realized by the following technical solutions:

[0009] The application provides a traditional Chinese medicine composition of Jinwu Goutong, which is composed of active ingredients of dog spine, epimedium, radix codonopsis, psoralea, papaya, curcuma, radix puerariae, radix et rhizoma bidentatae and amphiesma, wherein the active ingredient of dog spine is protocatechuic acid; the active ingredient of epimedium is icariin and icariin; the active ingredient of radix codonopsis is codonoloside; the active ingredient of psoralea is isopsoralen; the active ingredient of papaya is papaya polyphenol; the active ingredient of curcuma is curcumin; the active ingredient of radix puerariae is puerarin; the active ingredient of radix et rhizoma bidentatae is dehydroepiandrosterone; and the active ingredient of amphiesma is pulchellin.

[0010] Further, the traditional Chinese medicine composition contains, by weight, 35.8-40.3 mg of protocatechuic acid, 12.0-13.5 g of icariin, 7.2-8.1 g of icariin, 3.8-4.3 g of codonoloside, 1.2-1.35 mg of isopsoralen, 0.9-1.02 g of papaya polyphenol, 144-162 mg of curcumin, 1.04-1.17 g of puerarin, 125.4-141.1 mg of dehydroepiandrosterone, 320-360 mg of pulchellin and 115-125 g of amphiesma.

[0011] Still further, the traditional Chinese medicine composition contains, by weight, 38 mg of protocatechuic acid, 12.75 g of icariin, 7.65 g of icariin, 4.08 g of codonoloside, 1.275 mg of isopsoralen, 0.97 g of papaya polyphenol, 153 mg of curcumin, 1.1 g of puerarin, 133.2 mg of dehydroepiandrosterone, 340 mg of pulchellin and 120 g of amphiesma.

[0012] Still further, the preparation method of the traditional Chinese medicine composition comprises the following steps:

[0013] (1) protocatechuic acid, icariin, icariin, codonoloside, isopsoralen, papaya polyphenol, curcumin, puerarin, dehydroepiandrosterone and pulchellin are weighed according to the prescription amount, and then mixed uniformly, and the mixture is prepared for use;

[0014] (2) amphiesma is decocted with water for three times, each time for 2 hours, the decoction is combined, filtered, and the filtrate is concentrated to a clear extract with a relative density of 1.10-1.20 (80℃), and then cooled and prepared for use;

[0015] (3) the mixture obtained in steps (1) and (2) is added to the excipient to prepare a slurry, and then the slurry is subjected to boiling granulation and drying, and the temperature is controlled at 60-80℃, so as to obtain a dry mixture prepared for use;

[0016] (4) the dry mixture obtained in step (3) is sieved through a 80-100 mesh sieve, and then filled into a hollow capsule, or tabletted after granulation, or granulated after granulation, so as to obtain the traditional Chinese medicine composition.

[0017] The application provides the capsule, tablet and granule prepared by the above method.

[0018] The raw material and effective component used in the application have the following pharmacological properties:

[0019] Cibotium barometz (L.) J. Sm. has the functions of expelling wind and dampness, tonifying liver and kidney and strengthening waist and knees. It is recorded in Shennong Bencao Jing that it can treat waist and back weakness, joint pain, knee pain caused by dampness and cold and benefit old people. Cibotium barometz (L.) J. Sm. contains tannins, protocatechuic acid, volatile oil, pterin, aromatic compounds, drynaria alcohol, kaempferol and other chemical components, has the effects of anti-inflammatory analgesia, anti-rheumatism, anti-osteoporosis, liver protection, anti-oxidation, anti-cancer, treating ulcer, promoting blood circulation and stopping bleeding, anti-platelet aggregation and protecting brain nerves. According to the relevant literature, the average content of protocatechuic acid in Cibotium barometz (L.) J. Sm. is about 0.028%.

[0020] Herba Epimedii is listed in Shennong Bencao Jing as a medium-grade medicinal herb, has the effects of tonifying kidney yang, strengthening muscles and bones and expelling wind and dampness, is used for treating impotence and spermatorrhea caused by kidney yang deficiency, soft muscles and bones, rheumatism and arthralgia, numbness and spasm, and is a commonly used traditional Chinese medicine in clinic. Herba Epimedii contains icariin, icariin A, icariin C, icariin E, cycloartem, mannose, rhamnose, glucose, berberine, alkaloid A and other chemical components, can increase cardiovascular and cerebrovascular blood flow, promote hematopoietic function, immune function and bone metabolism, has the effects of tonifying kidney and strengthening yang and anti-aging. Icariin and icariin are the main active components of Herba Epimedii, have the effects of anti-oxidation, improving reproductive function, regulating bone metabolism, enhancing immunity and anti-inflammation. According to the relevant literature, the content of icariin in Herba Epimedii is 75.31 mg / g, and the content of icariin is 45.13 mg / g.

[0021] Campanumoea javanica Bl. subsp. javanica and Campanumoea javanica Bl. has the functions of tonifying middle energizer and replenishing qi, invigorating spleen and lung. According to modern medical analysis, the main components of Codonopsis pilosula include volatile oil, palmitic acid, alpha-pinene, borneol, syringin, codonopsis glycoside, choline, ryegrass alkaloid, sterol, acetate, friedelin, syringaldehyde, vanillic acid, 5-hydroxymethylfurfural and the like. According to the relevant literature, the content of active component codonopsis glycoside in Codonopsis pilosula is 36-73 mg / g.

[0022] Psoralea is a one-year-old herbaceous plant of the legume Psoralea, branches are hard, sparse white hair, with obvious gland points. It has the effects of invigorating deficiency, moistening lung, strengthening sinew and activating collateral, and is used for treating lumbar muscle strain, rheumatoid arthritis, treating lung heat, lung deficiency cough, pulmonary tuberculosis, chronic bronchitis, chronic hepatitis, spermatorrhea, leukorrhea. "Medicine Nature" records "main men's waist pain, cold and wet, all cold and stubborn, stop urination, cold in the abdomen". The main chemical components of Psoralea are psoralen, psoralen, psoralen, psoralen, psoralen, psoralen, psoralen, etc. According to the literature, isopsoralen can significantly promote the proliferation of MCF-7 cells, significantly increase the proliferation index of MCF-7 cells, and significantly increase the proportion of S phase cells. According to the relevant literature, the content of active ingredient isopsoralen in Psoralea is about 7.98-11.7mg / g.

[0023] Pseudocydonia sinensis (Thouin) C. K. Schneid. is a shrub or small tree of Rosaceae Pseudocydonia. Pseudocydonia has the effects of invigorating spleen, digesting, anti-inflammatory, protecting blood vessels, promoting breast development, containing vitamin C, carotene, papain, papain, malic acid, citric acid, tartaric acid, and polyphenol, etc. It has the effects of anti-oxidation, anti-aging, removing free radicals in the body, increasing skin elasticity and reducing wrinkle formation, and removing black pigment deposition in the body. It can improve human immunity, regulate endocrine function, and enhance constitution. Pseudocydonia can treat swelling of feet and muscle stiffness. According to the relevant literature, the content of active ingredient polyphenol in Pseudocydonia is about 8.55-10.74mg / g.

[0024] Curcuma longa L. is also known as Yujin, Baodingxiang, Haiming, and Huangjiang. It is a perennial herb of Zingiberales, Zingiberaceae, and Curcuma. Curcuma can promote blood circulation and remove blood stasis, and can also relieve pain. The main chemical components of Curcuma are curcumin, curcumin, gingerene, ar-turmerone, ar-turmerone, stigmasterol, sitosterol, monoene acid, diene acid, etc. It is used for treating chest and abdominal distension, shoulder and arm pain, intractable heart pain, postpartum blood pain, initial tinea, irregular menstruation, amenorrhea, and contusion. According to the relevant literature, the content of active ingredient curcumin in Curcuma is 1.5mg / g.

[0025] Radix Puerariae Radix Puerariae is the dried root of Pueraria lobata or Pueraria thomsonii, which belongs to Leguminosae. It has the functions of clearing heat and detoxifying, generating fluid and relieving thirst, tonifying kidney and spleen, benefiting stomach and tranquilizing mind, clearing heart and eyesight, nourishing skin, moistening intestines and defecation, and dispelling alcohol. Radix Puerariae contains daidzin, daidzein, glycitin, puerarin, puerarin-7-xyloside, etc. Puerarin is one of the main effective components of total flavonoids in Radix Puerariae, which has the functions of protecting blood vessels, reducing blood sugar and blood lipid, preventing and treating tumors, and regulating immunity. According to the relevant literature, the content of Radix Puerariae in terms of puerarin in each capsule of Jinwugutong capsules should not be less than 1.3 mg.

[0026] Achyranthes aspera L. is a plant of Achyranthes aspera L. in Amaranthaceae. The root is used for clearing heat and detoxifying, and has the functions of diuresis, and is used for treating cold, tonsillitis, diphtheria, mumps, urinary stones, nephritis edema, etc. The main chemical components of Achyranthes aspera L. are achyranthoside, oleanolic acid, achyranthesterone, sitosterol, achyranthesterone, rutin, isoquercitrin, kaempferol, etc. Achyranthesterone is one of the active components for promoting the proliferation of osteoblast-like cells in Achyranthes aspera L. According to the relevant literature, the content of achyranthesterone in Achyranthes aspera L. is about 0.1306%.

[0027] Clematis chinensis Osbeck is a perennial woody vine of Ranunculaceae and Clematis. It has the functions of dispelling wind and dampness, dredging collaterals and relieving pain, and is used for treating rheumatism, gout, limb numbness, muscle and tendon spasm, difficulty in flexing and extending, bone pharyngeal swelling, etc. Clematis chinensis Osbeck is rich in clematichinenoside, clematichinensin, anemonin, ranunculin, palmitic acid, linoleic acid, etc. Modern pharmacological research shows that Clematis chinensis Osbeck has the functions of analgesia, cholagogue, antibiosis, resisting histamine excitement, and antidiuresis, and also has the functions of inducing labor, relaxing smooth muscle, and making local muscle relaxed and promoting bone spur to fall off. According to the relevant literature, the content of anemonin in Clematis chinensis Osbeck is about 0.25%.

[0028] Zaocys dhumnades, also known as black snake, green snake and black flower snake, is a snake of Colubridae, which has the functions of dispelling wind, dredging collaterals and relieving convulsion. It is used for treating rheumatism, numbness and spasm, hemiplegia, tetanus, leprosy, tinea, scrofula and sore. Zaocys dhumnades mainly contains protein, fat, snake serum, thiamine, riboflavin, nicotinic acid, vitamins, etc. The meat of Zaocys dhumnades can dispel wind and dampness, dredge collaterals, and is used for treating numbness of skin, bone and joint tuberculosis, tinea, leprosy, tetanus, and infantile paralysis.

[0029] The present application has at least the following advantages and beneficial effects:

[0030] In the process of combining Cibotium barometz, Epimedium, Codonopsis pilosula, Psoralea corylifolia, Chaenomeles speciosa, Curcuma longa, Pueraria lobata, Achyranthes bidentata, Clematis chinensis, and Zaocys dhumnades, this invention has discovered that a drug preparation composed of specific dosage ratios has good effects in dispelling wind and dampness, promoting blood circulation and unblocking collaterals. Furthermore, it has been found that the active ingredients extracted from Cibotium barometz, Epimedium, Codonopsis pilosula, Psoralea corylifolia, Chaenomeles speciosa, Curcuma longa, Pueraria lobata, Achyranthes bidentata, and Clematis chinensis have further clarified the therapeutic components of each formulation, providing an ideal foundation for the modernization of traditional Chinese medicine research. This makes the combination of formulations more effective in promoting qi circulation and removing blood stasis, anti-inflammatory and antibacterial, dispelling wind and unblocking collaterals, and relieving spasms. The traditional Chinese medicine composition of this invention utilizes the effective monomeric components of medicinal materials to enhance the effects of dispelling wind and dampness, promoting blood circulation and unblocking collaterals, relieving lower back and leg pain and limb numbness caused by wind-cold-dampness syndrome, and treating osteoporosis. Through rational combination, the monomeric components promote each other's absorption and distribution, strengthen the synergistic interaction between several monomers, and have a stronger additive effect than individual drugs, thereby maximizing the therapeutic effect without drug dependence or toxic side effects. It achieves better therapeutic effects in treating lower back and leg pain, limb numbness, and preventing osteoporosis. Attached Figure Description

[0031] Figure 1 Microscopic images showing the effects of Jinwu Gutong capsules on the morphology of rat femoral tissue. Detailed Implementation

[0032] The pharmaceutical composition or preparation of the present invention is prepared by directly using extracts of traditional Chinese medicinal herbs such as Cibotium barometz, Epimedium, Codonopsis pilosula, Psoralea corylifolia, Chaenomeles speciosa, Curcuma longa, Pueraria lobata, Achyranthes bidentata, and Clematis chinensis, along with Zaocys dhumnades. Preferred active ingredients include protocatechuic acid from Cibotium barometz, icariin and icariin from Epimedium, codonopsis pilosula glycoside from Codonopsis pilosula, isopsoralen from Psoralea corylifolia, papain from Chaenomeles speciosa, curcumin from Curcuma longa, puerarin from Pueraria lobata, ecdysterone from Achyranthes bidentata, and anechoic acid from Clematis chinensis. Specific embodiments are as follows:

[0033] Examples 1-5: A Jinwu Gutong composition, as shown in Table 1, comprises the following components by weight:

[0034] Table 1:

[0035] Parts by weight Example 1 Example 2 Example 3 Example 4 Example 5 Protocatechuic acid 38 mg 38 mg 38 mg 38 mg 38 mg Icariin 12.75g 9g 18g 12g 12g Icaritin 7.65g 5g 10g 7g 7g Tangshenoside 4.08g 4.08g 4.08g 4.08g 4.08g Isopsoralen 1.275 mg 1.275 mg 1.275 mg 0.5 mg 3.5 mg Papain polyphenol 0.97g 0.97g 0.97g 0.97g 0.97g Curcumin 153 mg 153 mg 153 mg 153 mg 153 mg Puerarin 1.1g 1.1g 1.1g 1.1g 1.1g Ecdysterone 133.2 mg 133.2 mg 133.2 mg 133.2 mg 133.2 mg Anemonin 340 mg 340 mg 340 mg 340 mg 340 mg Rhabdophis 120g 120g 120g 120g 120g

[0036] Example 1: Preparation of the Golden Crow Bone-Strengthening Capsules of the Present Invention

[0037] Take 38mg of protocatechuic acid, 12.75g of icariin, 7.65g of icariin, 4.08g of tanshinone, 1.275mg of isopsoralen, 0.97g of papaya polyphenol, 153mg of curcumin, 1.1g of pueraria, 133.2mg of dehydroepiandrosterone, 340mg of anemarrhena, 120g of rinkhale, 350g of starch;

[0038] Preparation method: take each component by weight, mix evenly, boiling granulation drying, temperature control at 65℃, dry mixture 80 mesh crushing sieve, into 0 hollow capsule, get 1000 gold black bone capsule.

[0039] Specification: 0.35g / pill;

[0040] Usage: oral, 3 pills at a time, 3 times a day.

[0041] Example 2 Preparation of gold black bone capsule of the application (low dose of active ingredients of icariin)

[0042] Take 38mg of protocatechuic acid, 9g of icariin, 5g of icariin, 4.08g of tanshinone, 1.275mg of isopsoralen, 0.97g of papaya polyphenol, 153mg of curcumin, 1.1g of pueraria, 133.2mg of dehydroepiandrosterone, 340mg of anemarrhena, 120g of rinkhale, 350g of starch;

[0043] Preparation method: take each component by weight, mix evenly, boiling granulation drying, temperature control at 65℃, dry mixture 80 mesh crushing sieve, into 0 hollow capsule, get 1000 gold black bone capsule.

[0044] Specification: 0.35g / pill;

[0045] Usage: oral, 3 pills at a time, 3 times a day.

[0046] Example 3 Preparation of gold black bone capsule of the application (high dose of active ingredients of icariin)

[0047] Take 38mg of protocatechuic acid, 18g of icariin, 10g of icariin, 4.08g of tanshinone, 1.275mg of isopsoralen, 0.97g of papaya polyphenol, 153mg of curcumin, 1.1g of pueraria, 133.2mg of dehydroepiandrosterone, 340mg of anemarrhena, 120g of rinkhale, 200g of starch, 150g of dextrin;

[0048] Preparation method: take each component by weight, mix evenly, boiling granulation drying, temperature control at 65℃, dry mixture 80 mesh crushing sieve, whole grain after pressing, get 1000 gold black bone capsule.

[0049] Specification: 0.35g / pill;

[0050] Usage: orally, 3 pills at a time, 3 times a day.

[0051] Example 4 Preparation of Jinwu Guttong Granules of the Invention (Low Dose of Psoralen Active Ingredient)

[0052] Take 38 mg of protocatechuic acid, 12.75 g of icariin, 7.65 g of icariin, 4.08 g of tanshinone, 0.5 mg of isopsoralen, 0.97 g of papaya polyphenol, 153 mg of curcumin, 1.1 g of puerarin, 133.2 mg of dehydroepiandrosterone, 340 mg of anemonin, 120 g of rinkhale, and 5000 g of dextrin.

[0053] Preparation method: take each ingredient according to the amount, mix uniformly, and dry the granules by boiling at a temperature of 65℃. Crush the dry mixture to 80 mesh, sieve the dry mixture, and obtain 1000 packs of Jinwu Guttong Granules.

[0054] Specification: 5 g / pack;

[0055] Usage: orally, 3 packs at a time, 3 times a day.

[0056] Example 5 Preparation of Jinwu Guttong Capsules of the Invention (High Dose of Psoralen Active Ingredient)

[0057] Take 38 mg of protocatechuic acid, 12.75 g of icariin, 7.65 g of icariin, 4.08 g of tanshinone, 3.5 mg of isopsoralen, 0.97 g of papaya polyphenol, 153 mg of curcumin, 1.1 g of puerarin, 133.2 mg of dehydroepiandrosterone, 340 mg of anemonin, 120 g of rinkhale, and 350 g of starch.

[0058] Preparation method and usage method are the same as in Example 1.

[0059] Specification: 0.35 g / pill;

[0060] Usage: orally, 3 pills at a time, 3 times a day.

[0061] Experimental Example 1, Raw Material Prescription, Raw Material Dosage Screening, and Comparative Study

[0062] During the research stage of the present application, the preparation process of the original Jinwugutong capsule was studied. It was found that on the basis of the original preparation process, Puerariae radix and Curcumae radix were dried, crushed into fine powder, and sieved through an 80-mesh sieve. The fine powder was reserved. Eight kinds of ingredients including Dog spine, Herba Epimedii, Radix Codonopsis, Fructus Chaenomelis, Radix et Rhizoma Polygoni, Radix et Rhizoma Bidentatae, Radix et Rhizoma Clematidis, and Agkistrodon were decocted with 5 times the amount of water for three times, each time for 2 hours. The decoction was combined and concentrated into a clear paste. After cooling, the above fine powder was added. The active ingredients had ideal transfer rate and flow extract yield. The content of each active ingredient in the capsule was detected. The content range of protocatechuic acid, icariin, icariin, radix ginseng, isopsoralen, papaya polyphenol, curcumin, puerarin, ecdysterone, and anemonin was 0.015%-0.035% of crude drug, 74-76 mg / g of crude drug, 44-46 mg / g of crude drug, 35-45 mg / g of crude drug, 8.0-10.0 mg / g of crude drug, 8.5-10.5 mg / g of crude drug, 1.0-2.0 mg / g of crude drug, 3.1-4.2 mg / g of crude drug, 0.1%-0.2% of crude drug, and 0.1%-0.3% of crude drug, respectively. Therefore, further research on the prescription composition and dosage was carried out.

[0063] Comparative sample 1 (lacking active ingredient group of Herba Epimedii): The prescription was protocatechuic acid, radix ginseng, isopsoralen, papaya polyphenol, curcumin, puerarin, ecdysterone, anemonin, and Agkistrodon. The dosage and preparation method of Example 1 were used to prepare the composition comparative sample 1.

[0064] Comparative sample 2 (lacking active ingredient group of Fructus Chaenomelis): The prescription was protocatechuic acid, icariin, icariin, radix ginseng, papaya polyphenol, curcumin, puerarin, ecdysterone, anemonin, and Agkistrodon. The same dosage and preparation method as Example 1 were used to prepare the comparative sample 2.

[0065] Comparative sample 3 (isopsoralen was replaced by psoralen): The prescription was protocatechuic acid, icariin, icariin, radix ginseng, psoralen, papaya polyphenol, curcumin, puerarin, ecdysterone, anemonin, and Agkistrodon. The same dosage and preparation method as Example 1 were used to prepare the comparative sample 3.

[0066] Comparative sample 4 (puerarin was replaced by daidzin): The prescription was protocatechuic acid, icariin, icariin, radix ginseng, isopsoralen, papaya polyphenol, curcumin, daidzin, ecdysterone, anemonin, and Agkistrodon. The same dosage and preparation method as Example 1 were used to prepare the comparative sample 4.

[0067] Comparative sample 5: Jinwugutong capsule prepared from traditional Chinese medicinal raw materials.

[0068] Comparative samples 1-5: A kind of Jinwugutong composition, as shown in Table 2, including components in weight parts:

[0069] Table 2:

[0070]

[0071] Method: The above-mentioned comparative samples 1-4 were prepared according to the method of the embodiment of the present application, wherein the comparative samples 1-2 lack icariin, icariin, and isobavachin, and there are certain differences in the color and smell of the prepared medicines, wherein the comparative samples 3-4 replace isobavachin or puerarin, and there is no significant difference in the color and smell of the prepared medicines. Further long-term toxicity study experiments were carried out.

[0072] Experimental Example Two, Long-term Toxicity Study:

[0073] 1. The influence of the sample of the embodiment of the present application on the organ coefficients of rats

[0074] (1) Medicine: The sample 1-5 of the embodiment prepared by the present application, the comparative sample 1-5, and the blank group.

[0075] (2) Test object: 120 SPF rats, weighing 180-220 g, half male and half female, purchased from Liaoning Changsheng Biotechnology Co., Ltd., license number: SCXK(Liaoning)2020-0001, animal qualification certificate number: 210726210101193026. After purchasing the rats, they were adaptively fed for 1 week before the experiment. Rats were housed in cages at 5 per cage, room temperature (20-25°C), RH 40%-70%, light and dark for 12 h each, and the animals were free to drink water and eat food during the experiment.

[0076] (3) Dosing time: The above groups were dosed according to the relevant dose in units of weeks, with continuous gavage for 6 days and 1 day of drug withdrawal for treatment, for 12 weeks. After the last dose, the animals were fed normally for 4 weeks.

[0077] (4) General condition observation and recording: daily observation and recording of appearance, behavior, gland secretion, respiration, feces, food intake, body weight, and local reaction to administration, weekly determination of body weight and recording of food intake.

[0078] (5) Animal disposal: After the last dose or normal diet and water recovery feeding for 4 weeks, the body weight of each group was recorded, and the animals were fasted for 12 h without water, blood was collected from the abdominal aorta, and all animals were sacrificed, systemically dissected, and the weights of heart, liver, spleen, lung, kidney, adrenal gland, thymus, brain, testis, epididymis, ovary, uterus, etc. were measured and histopathological examination of liver and kidney tissues was performed, the organ coefficients were calculated, and blood biochemical indicators (AST, ALT, BUN, CREA, ALP, TBIL) were detected.

[0079] (6) Blood biochemical index detection: according to the "Guiding Principles for Clinical Evaluation of Traditional Chinese Medicine Drug-induced Liver Injury", the biochemical indexes such as AST, ALT, BUN, CREA, ALP, TBIL, etc. are detected.

[0080] Table 3: Comparison of the influence of the sample of the embodiment of the present application on the organ coefficients of rats

[0081]

[0082] Table 3 (continued)

[0083]

[0084] The results show that: (1) Comparison of the sample of the embodiment: compared with the blank group, the organ coefficients of rats in the sample 1 (prepared according to the prescription and dosage screened according to the present application) had no significant change (p<0.05). The liver coefficient of the sample 2 (low-dose group of icariin and epimedium-derived active ingredients) increased slightly (p<0.05). The kidney coefficient of the sample 3 group (high-dose group of icariin and epimedium-derived active ingredients) increased (p<0.05), and the ovary coefficient decreased (p<0.05). The organ coefficients of rats in the sample 4 group (low-dose of isopsoralen) had no obvious change (p<0.05), and the liver, kidney and testis coefficients of the sample 5 group (high-dose of isopsoralen) increased (p<0.05), as shown in Table 3.

[0085] (2) Comparison of the comparative sample: compared with the blank group, the testis and ovary coefficients of the comparative sample 1 (lacking icariin and epimedium-derived active ingredients) and the comparative sample 2 (lacking isopsoralen) decreased significantly (p<0.05), the liver coefficient of the comparative sample 3 (isopsoralen replaced by psoralen) increased (p<0.05), the liver, lung and testis coefficients of the comparative sample 4 (daidzin replaced by puerarin) did not increase (p<0.05), and the relevant coefficients of the comparative sample 5 (existing medicine Jinwugutong Capsules prepared from traditional Chinese medicinal materials) did not change significantly (p<0.05).

[0086] (3) Recovery period: after the last administration, the rats were fed normally for 4 weeks, and there was no significant difference in the organ coefficients compared with the blank group.

[0087] The experiment studies the safety of Jinwugutong Capsules under repeated administration, observes whether the test animals have liver and kidney toxicity, main manifestations, affected organs and recovery after drug withdrawal. Based on the Technical Guidelines for Long-term Toxicity Research of Traditional Chinese Medicine and Natural Medicine, the long-term toxicity experiment of normal rats is conducted for 90 days to clarify the toxicity, main manifestations, affected organs and recovery of long-term use of Jinwugutong Capsules. In summary, under the conditions of this experiment, Jinwugutong Capsules did not cause obvious long-term toxicity. At a clinical dose of 20 times, Jinwugutong Capsules had no liver and kidney toxicity on experimental rats.

[0088] Disintegration time limit: 3 capsules of the prepared sample in Example 1 were placed in beakers containing 200 ml of water (water temperature was 15-25°C), and many bubbles were released. The capsules were completely dissolved in water, and no aggregated particles remained. Each capsule disintegrated within 30 minutes, meeting the relevant standards.

[0089] Dose difference: 10 capsules of the prepared sample in Example 1 were precisely weighed for the content and shell according to the standard method in the Pharmacopoeia. The dose difference was less than 10% compared with the labeled dose, meeting the relevant requirements.

[0090] Stability: Through the accelerated stability experiment, 1 capsule of each sample prepared in Examples 1-5 was randomly selected and placed at a temperature of 40°C ± 2°C and a relative humidity of 75% ± 5%. The samples were taken at the end of the first month, second month, third month, and sixth month of the experiment for examination. The results showed that there were no abnormalities in the examination items, and the content of the active ingredient did not change significantly, meeting the quality standard requirements.

[0091] Experimental Example Four, Efficacy Study

[0092] (1) Animals: 80 SPF non-pregnant female SD rats, weighing about 250-300g, purchased from the Animal Institute of Guizhou University of Chinese Medicine (Certificate No. 10662350000051 Animal License No.: SCXK(Jiang)2021-0003).

[0093] (2) Drugs: Jinwugutong Capsules prepared by the present application, original product Jinwugutong Capsules (Guizhou Shengshi Longfang Pharmaceutical Co., Ltd.), alendronate sodium tablets (Hangzhou Merck Dong Pharmaceutical Co., Ltd.), and lipopolysaccharide (Shanghai Yuan Ye Biological Technology Co., Ltd.).

[0094] (3) Animal modeling: 80 SPF female rats, after adaptive feeding for one week, 10% chloral hydrate 0.3 mL / 100 g intraperitoneal injection, after the rats were anesthetized, the skin and muscle were cut along the dorsal lumbar vertebrae 1 cm from the midline, the abdominal cavity was entered, the ovary was clamped with a sterile forceps, the oviduct was ligated at the uterine horn, and the bilateral ovaries of the rats were removed. The modeling method of the sham operation group is the same, but the ovary is not removed, and the fat mass equal to the size of the ovary near the ovary is removed.

[0095] (4) Animal grouping: after the modeling is completed, 72 rats are normally fed for 12 weeks, and the rats are randomly divided into 9 groups: sham operation group (abbreviation: Sham), model group (abbreviation: OVX), alendronate sodium group (abbreviation: AS), lipopolysaccharide group (abbreviation: L), original product Jinwugutong capsule group (abbreviation: J), alendronate sodium + LPS immune stimulation group (abbreviation: LAS), Jinwugutong capsule + LPS immune stimulation group (abbreviation: LJ), Jinwugutong capsule + alendronate sodium tablet combined drug group (abbreviation: JAS), and Jinwugutong capsule prepared by the application (abbreviation: ZJ).

[0096] (5) Dose design

[0097] Jinwugutong capsule and alendronate sodium tablet dose: according to the equivalent conversion dose of human and animal homologous drugs, the administration dose of Jinwugutong capsule is 0.405 g / kg, and the administration dose of alendronate sodium tablet is 0.9 mg / kg.

[0098] Lipopolysaccharide dose: according to relevant literature, the lipopolysaccharide dose is designed as 0.1 mg / kg solution.

[0099] (6) Solution configuration

[0100] Lipopolysaccharide solution configuration: accurately weigh a certain amount of lipopolysaccharide powder, add an appropriate amount of normal saline, vortex for 30 minutes, prepare a 1 mg / mL concentrated stock solution; then take an appropriate amount of concentrated stock solution with a pipette, add 5 times the volume of normal saline, vortex for 15 minutes, prepare a 0.2 mg / mL secondary concentrated stock solution; then dilute the appropriate volume of secondary concentrated stock solution with normal saline by 4 times, vortex for 15 minutes, and prepare a 0.05 mg / mL lipopolysaccharide solution.

[0101] Alendronate sodium suspension solution configuration: grind the alendronate sodium tablet into powder, weigh an appropriate amount of alendronate sodium tablet powder, dissolve in warm water, and prepare a suspension solution for standby.

[0102] Jinwugutong capsule solution configuration: take Jinwugutong capsule, remove the capsule shell, weigh an appropriate amount of content, dissolve in warm water, and prepare a suspension solution for standby.

[0103] (7) Animal administration

[0104] LAS, AS group: intragastric administration of 0.9mg / kg alendronate sodium tablet suspension; LJ, J, ZJ group: intragastric administration of 0.405g / kg Jinwugutong capsule suspension; JAS group 0.9mg / kg alendronate sodium tablet suspension + 0.405g / kg Jinwugutong capsule suspension, the rest of the group intragastric administration of the same volume of distilled water, in units of weeks, 6 days of continuous intragastric administration per week, 1 day of drug withdrawal, for 4 weeks. After the last administration, fasting without water, 12h, L, LJ, LAS tail vein LPS solution, the rest of the group tail vein injection of normal saline, injection volume 20mL / kg, half an hour later, intragastric administration of corresponding drugs, 12h after the last administration, dissection.

[0105] (8) Specimen collection: at the 4th week of administration, the rats were fasted for 24h, and the urine was collected. The obtained urine was centrifuged at 3000r / min for 10min, and the separated urine was stored in a-20℃ refrigerator. After the last administration, the rats were fasted without water, and blood was collected from the abdominal vein after anesthesia, centrifuged, and the supernatant was taken. Six serum samples were selected for liver function index and bone homeostasis index detection. The liver, kidney, uterus, left tibia and femur of the rats were fixed in 10% paraformaldehyde.

[0106] (9) Determination of bone metabolism index: the contents of inorganic calcium, phosphorus and urine creatinine in urine and the contents of serum TRAP, E2, β-CTX, ALP and BGP were detected according to the instructions of the kit.

[0107] The microscopic graph of the influence of the prepared Jinwugutong capsule of the application on the femur tissue morphology of rats (see the description attached Figure 1 ) showed that compared with the Sham, the bone trabecula of the model group rats was obviously sparse, and the bone marrow cavity was significantly expanded; compared with the model group, the number of bone trabecula of the J group, the AS group and the ZJ group rats was obviously increased, arranged in order, connected closely, and the bone marrow cavity was small. Figure 1 After intragastric administration of Jinwugutong capsule in this study, Ca2+ / Cr and Pi / Cr were significantly down-regulated, indicating that Jinwugutong capsule can resist calcium and phosphorus loss in ovariectomized rats. The results of HE staining of rat femur tissue and serum biochemical results also showed that intragastric administration of Jinwugutong capsule can increase the number of bone trabecula and the tightness of connection, indicating that Jinwugutong capsule can resist ovariectomized osteoporosis.

[0108] (10) The influence of the prepared Jinwugutong capsule of the application on the liver function of rats in the treatment of osteoporosis was studied: the osteoporosis model of ovariectomized rats was replicated using 250g-300g SD rats, and the immune stress state of rats was induced by LPS. From the results of serum biochemistry, urine biochemistry, organ coefficient and histopathology, the specific changes are shown in Table 4.

[0109] Table 4 Change table of organ coefficient of rats in each group

[0110] Group Liver Kidney Uterus Sham 2.352±0.265 0.516±0.068 0.189±0.089 OVX 2.085±0.135 0.491±0.055 0.086±0.033 L 2.658±0.117 0.551±0.048 0.088±0.024 J 2.030±0.188 0.406±0.048 0.121±0.072 LJ 2.501±0.221 0.588±0.053 0.113±0.048 AS 2.432±0.240 0.524±0.085 0.132±0.104 LAS 2.409±0.220 0.665±0.036 0.132±0.167 JAS 2.172±0.225 0.475±0.045 0.114±0.084 ZJ 1.950±0.885 0.361±0.045 0.117±0.07

[0111] Results show that: compared with the Sham group: the liver coefficient of the L group is significantly higher than that of the Sham group (p<0.05), the kidney coefficient is higher than that of the Sham group, but there is no significant difference; the uterus coefficient of the OVX group is significantly smaller than that of the Sham group (p<0.05); compared with the J group: the liver and kidney coefficients of the LJ group are significantly higher than those of the J group (p<0.05); compared with the AS group: the kidney coefficient of the LAS group is increased, but there is no significant difference; the kidney coefficient of the J group is lower than that of the JAS group (p<0.05), compared with the J group: the kidney coefficient of the ZJ group is lower than that of the J group (p<0.05); compared with the OVX group: the uterus coefficient of each drug group is slightly higher than that of the OVX group, and there is no difference between groups, see Table 5. It can be seen that the prepared Jinwu Gutong capsules of the application can increase the serum BGP and E2 levels, and reduce the serum TRAP, β-CTX and ALP levels, and from the data, it can be seen that the prepared Jinwu Gutong capsules of the application resist the loss of calcium and phosphorus in ovariectomized rats, increase the trabecular bone number of ovariectomized rats to treat osteoporosis, and do not aggravate the liver damage of immune stress rats.

Claims

1. A traditional Chinese medicine composition for treating osteoporosis, comprising the active ingredients of Cibotium barometz, Epimedium, Codonopsis pilosula, Psoralea corylifolia, Chaenomeles speciosa, Curcuma longa, Pueraria lobata, Achyranthes bidentata, Clematis chinensis, and Zaocys dhumnades, wherein the active ingredient of Cibotium barometz is protocatechuic acid; the active ingredients of Epimedium are icariin and icariin; the active ingredient of Codonopsis pilosula is codonopsis glycoside; the active ingredient of Psoralea corylifolia is isopsoralen; the active ingredient of Chaenomeles speciosa is papain polyphenol; the active ingredient of Curcuma longa is curcumin; the active ingredient of Pueraria lobata is puerarin; the active ingredient of Achyranthes bidentata is ecdysterone; and the active ingredient of Clematis chinensis is anechoic acid. The above active ingredients, calculated by weight, contain protocatechuic acid 33.6-42.6 mg, icariin 11.25-14.25 g, icariin 6.75-8.55 g, codonopin 3.6-4.6 g, isopsoralen 1.125-1.425 mg, papaya polyphenols 0.85-1.08 g, curcumin 135-171 mg, puerarin 0.98-1.24 g, ecdysterone 117.5-148.8 mg, anestheticin 300-380 mg, and black sedge 110-130 g.

2. The traditional Chinese medicine composition according to claim 1, by weight, contains 35.8-40.3 mg of protocatechuic acid, 12.0-13.5 g of icariin, 7.2-8.1 g of icariin, 3.8-4.3 g of codonopsis glycoside, 1.2-1.35 mg of isopsoralen, 0.9-1.02 g of papain, 144-162 mg of curcumin, 1.04-1.17 g of puerarin, 125.4-141.1 mg of ecdysterone, 320-360 mg of anesthetic, and 115-125 g of Zaocys dhumnades.

3. The traditional Chinese medicine composition according to claim 2, by weight, contains 38 mg of protocatechuic acid, 12.75 g of icariin, 7.65 g of icariin, 4.1 g of codonopsis glycoside, 1.275 mg of isopsoralen, 0.97 g of papaya polyphenols, 153 mg of curcumin, 1.1 g of puerarin, 133.2 mg of ecdysterone, 340 mg of anesthetic, and 120 g of Zaocys dhumnades.

4. A Jinwu Gutong pharmaceutical preparation, made from the traditional Chinese medicine composition and excipients described in any one of claims 1-3.

5. A method for preparing the Jinwu Gutong pharmaceutical preparation as described in claim 4, characterized in that, Includes the following steps: (1) Weigh out the following ingredients according to the formula: protocatechuic acid, icariin, icariin, codonopsis glycoside, isopsoralen, papaya polyphenol, curcumin, puerarin, ecdysterone, and anechosamine, mix them evenly, and set the mixture aside. (2) Boil the black-striped snake in water three times, each time for 2 hours, combine the decoctions, filter, concentrate the filtrate to a clear paste with a relative density of 1.10-1.20 at 80℃, and let it cool for later use; (3) Add the mixture obtained in steps (1) and (2) to the auxiliary materials to make a slurry, and perform boiling granulation and drying. The drying temperature is controlled at 60-80℃ to obtain a dried mixture for later use. (4) Crush the mixture obtained from drying in step (3) into 80-100 mesh and sieve it, then fill it into empty capsules, or granulate the mixture obtained from drying in step (3) and compress it into tablets, or sieve the granules to obtain the final product.

6. The Jinwu Gutong pharmaceutical preparation according to claim 4, wherein the dosage form is capsule, tablet or granule.

7. The preparation method according to claim 5, characterized in that, The boiling granulation drying temperature in step (3) is 65°C.

8. The preparation method according to claim 5, characterized in that, The mixture obtained in step (4) is dried, pulverized at 80 mesh, and sieved.

9. The use of the traditional Chinese medicine composition according to any one of claims 1-3 in the preparation of a medicament for treating osteoporosis.

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