Traditional Chinese medicine composition for treating osteoporosis

Abstract

The invention provides a traditional Chinese medicine composition for treating osteoporosis. The traditional Chinese medicine composition comprises rhizoma drynariae, eucommia ulmoides, fructus psoraleae, ligusticum wallichii, raw radix astragali, codonopsis pilosula, radix cyathulae and radix achyranthis bidentatae. The traditional Chinese medicine composition can be used for effectively treating osteoporosis, myopenia-osteoporosis and climacteric osteoporosis.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of traditional Chinese medicine compositions, in particular to a traditional Chinese medicine composition for treating osteoporosis and a preparation method and application thereof. BACKGROUND

[0002] Sarcopenia-osteoporosis is a high-incidence comorbid syndrome in the elderly population, which is characterized by muscle mass reduction, muscle strength decline (sarcopenia) and bone density reduction, bone fragility increase (osteoporosis). The medical community initially regarded sarcopenia (SP) and osteoporosis (OP) as two independent diseases, but as the research on the interaction between muscle and bone deepened, more and more evidence showed that the occurrence of SP and other muscle diseases would increase the risk of OP and other bone diseases, and vice versa. The comorbidity of the two would further exacerbate the original negative health outcomes. Some scholars described this relationship as a "dangerous duo". The two promote each other through endocrine, mechanical and inflammatory pathways, forming a vicious cycle: muscle atrophy leads to reduced mechanical stimulation of the skeleton, accelerating bone loss; and increased bone fragility limits mobility, further exacerbating muscle degradation.

[0003] In 2009, Binkley et al. proposed the concept of "sarcopenia-osteoporosis", which first explicitly defined the relationship between sarcopenia and osteoporosis in terms of disease name. It was then proposed that sarcopenia and osteoporosis could be combined and treated as a "motor dysfunction syndrome". The concept of sarcopenia-osteoporosis is conducive to people's thinking about the nature of the interaction between muscle and bone from a holistic perspective, and will provide new ideas for the study of the musculoskeletal system.

[0004] Currently, the main treatments for sarcopenia-osteoporosis are exercise therapy and nutritional support. Exercise therapy can significantly increase muscle mass, muscle strength and improve physical function, and has a promoting effect on maintaining bone structure, improving bone density and reducing the risk of falls and fragile fractures. Exercise therapy needs to follow the principles of individualization and long-term adherence, but needs to avoid adverse events such as trauma caused by improper exercise. There are few drugs in Western medicine that target both osteoporosis and sarcopenia, and their efficacy is uncertain. Anti-bone resorption drugs such as bisphosphonates and RANKL inhibitors are mainly used, which can delay bone loss, but have limited effect on improving muscle function, and long-term use may cause gastrointestinal reactions, jaw necrosis and other side effects.

[0005] Existing traditional Chinese medicine products such as artificial tiger bone powder (Jintiange capsules), Gushukang capsules / granules, Xianlinggubao capsules, Zhuangguzitong capsules, etc. are all indicated for osteoporosis. There is no marketed traditional Chinese medicine product for treating sarcopenia-osteoporosis, and there is also no registered traditional Chinese medicine product for clinical trials. SUMMARY

[0006] The present application provides a traditional Chinese medicine composition, which can effectively treat osteoporosis, especially muscle-osteoporosis and menopausal osteoporosis. Application of the traditional Chinese medicine composition can effectively reduce pathological weight gain, improve grip strength, increase trabecular bone thickness, bone volume fraction and bone mineral density, reduce trabecular bone separation degree, and improve muscle structure and bone tissue structure. Further, the traditional Chinese medicine composition plays a role in bone protection by increasing the content of iridin; reduces bone turnover rate by reducing ALP in serum; and achieves the purpose of treating menopausal osteoporosis by increasing the amount of estrogen. In addition, through detection of serum markers, it is found that the traditional Chinese medicine composition can reduce bone resorption, inhibit osteoblast differentiation and inhibit osteoclast generation. The specific scheme is as follows:

[0007] In a first aspect of the present application, a traditional Chinese medicine composition is provided, which comprises rhizoma drynariae, eucommia ulmoides, psoralea corylifolia, chuanxiong rhizome, radix astragali, radix codonopsis, radix cyathulae and radix cyathulae.

[0008] Among them, the rhizoma drynariae, eucommia ulmoides and psoralea corylifolia are monarch drugs, which can tonify kidney and strengthen bone;

[0009] The radix astragali and radix codonopsis are minister drugs, which can tonify qi and invigorate the spleen, and tonify postnatal essence to strengthen the skeleton;

[0010] The radix cyathulae and radix cyathulae are auxiliary drugs, which can tonify liver and kidney, strengthen sinew and bone, and activate blood and unblock channels;

[0011] The chuanxiong rhizome is a guide drug, which can dredge bone nutrition pathway and promote bone growth.

[0012] Preferably, the monarch drugs are rhizoma drynariae, eucommia ulmoides and psoralea corylifolia.

[0013] Preferably, the traditional Chinese medicine composition in the present application comprises rhizoma drynariae, eucommia ulmoides, psoralea corylifolia, chuanxiong rhizome, radix astragali, radix codonopsis, radix cyathulae and radix cyathulae.

[0014] In a specific embodiment of the present application, the traditional Chinese medicine composition comprises, by mass fraction:

[0015] Rhizoma drynariae 8-20 parts, for example 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 parts, etc.; preferably 14-16 parts;

[0016] Eucommia ulmoides 10-20 parts, for example 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 parts, etc.; preferably 14-16 parts;

[0017] Psoralea corylifolia 5-15 parts, for example 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 parts, etc.; preferably 8-12 parts;

[0018] Chuanxiong 10-20 parts, for example 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 parts, etc.; preferably 14-16 parts;

[0019] Huangqi 20-40 parts, for example 20, 25, 30, 35, or 40 parts, etc.; preferably 25-35 parts;

[0020] Dangshen 20-40 parts, for example 20, 25, 30, 35, or 40 parts, etc.; preferably 25-35 parts;

[0021] Chuanniuxie 10-20 parts, for example 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 parts, etc.; preferably 14-16 parts; and

[0022] Huaijiuxie 10-20 parts, for example 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 parts, etc.; preferably 14-16 parts. In one specific embodiment of the present application, the Chinese medicine ingredients in the Chinese medicine composition, by mass parts, include or are:

[0023] Gushu bu 8-20 parts, for example 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 parts, etc.; preferably 14-16 parts;

[0024] Duzhong 10-20 parts, for example 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 parts, etc.; preferably 14-16 parts;

[0025] Buguzhi 5-15 parts, for example 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 parts, etc.; preferably 8-12 parts;

[0026] Chuanxiong 10-20 parts, for example 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 parts, etc.; preferably 14-16 parts;

[0027] Huangqi 20-40 parts, for example 20, 25, 30, 35, or 40 parts, etc.; preferably 25-35 parts;

[0028] Dangshen 20-40 parts, for example 20, 25, 30, 35, or 40 parts, etc.; preferably 25-35 parts;

[0029] Chuanniuxie 10-20 parts, for example 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 parts, etc.; preferably 14-16 parts; and

[0030] Drynaria fortunei 10-20 parts, for example 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 parts, etc.; preferably 14-16 parts.

[0031] In one embodiment of the present application, the traditional Chinese medicine composition comprises, by mass parts, Drynaria fortunei 10-20 parts, Eucommia ulmoides Oliv. 10-20 parts, Psoralea corylifolia 8-12 parts, Ligusticum chuanxiong Hort. 10-20 parts, Astragalus membranaceus var. mongholicus 25-35 parts, Codonopsis pilosula 25-35 parts, Radix Cyathulae 10-20 parts, and Radix Hedysari 10-20 parts.

[0032] In one embodiment of the present application, the traditional Chinese medicine composition comprises, by mass parts, Drynaria fortunei 10-20 parts, Eucommia ulmoides Oliv. 10-20 parts, Psoralea corylifolia 8-12 parts, Ligusticum chuanxiong Hort. 10-20 parts, Astragalus membranaceus var. mongholicus 25-35 parts, Codonopsis pilosula 25-35 parts, Radix Cyathulae 10-20 parts, and Radix Hedysari 10-20 parts.

[0033] In one embodiment of the present application, the traditional Chinese medicine composition comprises, by mass parts, Drynaria fortunei 10-20 parts, Eucommia ulmoides Oliv. 10-20 parts, Psoralea corylifolia 8-12 parts, Ligusticum chuanxiong Hort. 10-20 parts, Astragalus membranaceus var. mongholicus 25-35 parts, Codonopsis pilosula 25-35 parts, Radix Cyathulae 10-20 parts, and Radix Hedysari 10-20 parts.

[0034] In one embodiment of the present application, the traditional Chinese medicine composition comprises, by mass parts, Drynaria fortunei 10-20 parts, Eucommia ulmoides Oliv. 10-20 parts, Psoralea corylifolia 8-12 parts, Ligusticum chuanxiong Hort. 10-20 parts, Astragalus membranaceus var. mongholicus 25-35 parts, Codonopsis pilosula 25-35 parts, Radix Cyathulae 10-20 parts, and Radix Hedysari 10-20 parts.

[0035] The Drynaria fortunei is the dried rhizome of Drynaria fortunei of the family Drynariaceae.

[0036] The Eucommia ulmoides Oliv. is the dried bark of Eucommia ulmoides Oliv. of the family Eucommiaceae.

[0037] The Psoralea corylifolia is the dried mature fruit of Psoralea corylifolia of the family Leguminosae.

[0038] The Ligusticum chuanxiong Hort. is the dried rhizome of Ligusticum chuanxiong Hort. of the family Umbelliferae.

[0039] The Astragalus membranaceus var. mongholicus is the dried root of Astragalus membranaceus var. mongholicus of the family Leguminosae.

[0040] The Codonopsis pilosula is the dried root of Codonopsis pilosula of the family Cucurbitaceae.

[0041] The said Cyathula is the dried root of Cyathula officinalis Kuan.

[0042] The said Cyathula is the dried root of Cyathula officinalis Kuan.

[0043] Preferably, the said Eucommia ulmoides is salt Eucommia ulmoides.

[0044] Further preferably, the said salt Eucommia ulmoides is salt Eucommia ulmoides prepared by frying broken silk with salt water.

[0045] Preferably, the said Fructus Psoraleae is salt Fructus Psoraleae.

[0046] Further preferably, the said salt Fructus Psoraleae is salt Fructus Psoraleae evenly mixed with salt, slightly moistened, and fried in a pan with a low fire until slightly puffed.

[0047] Preferably, the said Drynariae Rhizoma is scalded Drynariae Rhizoma.

[0048] Further preferably, the said scalded Drynariae Rhizoma is Drynariae Rhizoma prepared by sand scalding.

[0049] Preferably, the dosage form of the said traditional Chinese medicine composition includes tablets, capsules, powders, pills, granules, injections, decoctions, wine, ointments, pastes, plasters, or oral liquids.

[0050] The said traditional Chinese medicine composition further includes pharmaceutically acceptable excipients.

[0051] Preferably, the said excipients include one or more of diluents, disintegrants, lubricants, excipients, binders, glidants, fillers, or surfactants.

[0052] Further preferably, the said diluents include but are not limited to one or more of lactose, starch, pre-gelatinized starch, microcrystalline cellulose, sorbitol, mannitol, or inorganic calcium salt.

[0053] Further preferably, the said disintegrants include but are not limited to one or more of cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, alginic acid, or microcrystalline cellulose.

[0054] Further preferably, the said lubricants include but are not limited to one or more of stearic acid, sodium stearate, magnesium stearate, calcium stearate, polyethylene glycol, talc, or hydrogenated vegetable oil.

[0055] Further preferably, the said excipients include one or more of fructose, galactose, mannitol, polyvinyl alcohol, sodium carboxymethyl cellulose, erythritol, or cyclodextrin.

[0056] Further preferably, the binder includes, but is not limited to, one or more than two of starch paste, hydroxypropyl methylcellulose, and polyvinylpyrrolidone.

[0057] Further preferably, the glidant includes, but is not limited to, one or more than two of microfine silica, talc, or magnesium trisilicate.

[0058] Further preferably, the filler includes one or more than two of sucrose, bifidus sugar, maltodextrin, silicon dioxide, calcium carbonate, magnesium carbonate, aluminum powder, or phenolic resin.

[0059] Further preferably, the surfactant includes one or more than two of Tween-80, cetyltrimethylammonium bromide, sodium stearyl sulfate, or sodium lauryl sulfate.

[0060] The traditional Chinese medicine composition can treat osteoporosis.

[0061] The traditional Chinese medicine composition can treat sarcopenia-osteoporosis.

[0062] The traditional Chinese medicine composition can treat menopausal osteoporosis.

[0063] Preferably, the treatment of osteoporosis includes taking 1-5 (e.g., 2, 3, 4, 4.5, 5, etc.) g / kg of crude drug per day, and taking the traditional Chinese medicine composition for 1-24 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24, preferably 2-6) months.

[0064] In a second aspect, the present application provides a preparation method of the traditional Chinese medicine composition of the first aspect, which includes powdering, water decocting, stir-frying, steeping, percolation, supercritical extraction, or reflux extraction.

[0065] Preferably, the water decocting is performed at least once, e.g., 1, 2, 3, 4, or 5 times.

[0066] Preferably, the water decocting is performed at least once, e.g., 1, 2, 3, 4, or 5 times.

[0067] Preferably, the water decocting is performed at least once, e.g., 1, 2, 3, 4, or 5 times.

[0068] Preferably, the water decocting is performed at least once, e.g., 1, 2, 3, 4, or 5 times.

[0069] Preferably, the water decocting is performed at least once, e.g., 1, 2, 3, 4, or 5 times.

[0070] Preferably, the time for each water decoction is 0.5-2h.

[0071] Preferably, the obtained medicinal liquid is concentrated to 2-4g / mL.

[0072] In one specific embodiment of the present application, the preparation method comprises adding water with a volume of 8-12 times the total mass of the ingredients of the traditional Chinese medicine composition, decocting for 0.5-2h, filtering the medicinal liquid while hot, adding water with a volume of 5-10 times to the medicinal residue and decocting for another 0.5-2h, combining the two filtrates, and concentrating the combined filtrate to a medicinal liquid with a concentration of 2-4g / mL.

[0073] In a third aspect of the present application, the traditional Chinese medicine composition of the first aspect or the traditional Chinese medicine composition obtained by the preparation method of the second aspect is used in the preparation of a drug for treating a disease.

[0074] The disease is osteoporosis.

[0075] Preferably, the disease is sarcopenia-osteoporosis or menopausal osteoporosis.

[0076] Preferably, the treatment of the disease comprises administering an effective amount of the traditional Chinese medicine composition to a subject in need thereof.

[0077] Preferably, the traditional Chinese medicine composition can effectively reduce pathological weight gain, improve grip strength, increase trabecular bone thickness, bone volume fraction, bone mineral density, reduce trabecular bone separation degree, and improve muscle structure and bone tissue structure.

[0078] Further preferably, the muscle structure is skeletal muscle structure, such as the skeletal muscle structure of the limbs.

[0079] Further preferably, the bone tissue structure is femur structure.

[0080] Preferably, the traditional Chinese medicine composition plays a role in bone protection by increasing the content of iridin.

[0081] Preferably, the traditional Chinese medicine composition reduces the bone turnover rate by reducing ALP in serum.

[0082] Preferably, the traditional Chinese medicine composition achieves the purpose of treating menopausal osteoporosis by increasing the amount of estrogen.

[0083] The traditional Chinese medicine composition can reduce bone resorption, inhibit osteoblast differentiation, and inhibit osteoclastogenesis.

[0084] In a fourth aspect of the present application, the traditional Chinese medicine composition of the first aspect or the traditional Chinese medicine composition obtained by the preparation method of the second aspect is used, and the use comprises:

[0085] A) use in the manufacture of a product for reducing pathological weight gain, increasing grip strength;

[0086] B) use in the manufacture of a product for increasing trabecular bone thickness, bone volume fraction and or bone mineral density, and / or reducing trabecular separation;

[0087] C) use in the manufacture of a product for improving muscle structure and bone tissue structure;

[0088] D) use in the manufacture of a product for increasing iridin in serum;

[0089] E) use in the manufacture of a product for reducing serum alkaline phosphatase (ALP) in serum;

[0090] F) use in the manufacture of a product for increasing the amount of estrogen; or,

[0091] G) use in the manufacture of a product for reducing bone resorption, inhibiting osteoblast differentiation and inhibiting osteoclastogenesis.

[0092] Preferably, the muscle structure is skeletal muscle structure, for example, skeletal muscle structure of the limbs.

[0093] Preferably, the bone tissue structure is femur structure.

[0094] Preferably, the product is a medicament.

[0095] In a fifth aspect, the present application provides a method for treating and / or preventing osteoporosis or sarcopenia-osteoporosis, the method comprising administering to a subject in need thereof an effective amount of the traditional Chinese medicine composition of the first aspect or the traditional Chinese medicine composition obtained by the preparation method of the second aspect.

[0096] In a sixth aspect, the present application provides a method for treating and / or preventing menopausal osteoporosis, the method comprising administering to a subject in need thereof an effective amount of the traditional Chinese medicine composition of the first aspect or the traditional Chinese medicine composition obtained by the preparation method of the second aspect.

[0097] In a seventh aspect, the present application provides a method for improving muscle structure and bone tissue structure, the method comprising administering the traditional Chinese medicine composition of the first aspect or the traditional Chinese medicine composition obtained by the preparation method of the second aspect.

[0098] The term "treatment" as used herein refers to slowing, interrupting, halting, controlling, stopping, reducing, or reversing the progression or severity of a sign, symptom, disorder, condition, or disease, after the disease has begun to develop, but does not necessarily involve the complete elimination of all disease-related signs, symptoms, conditions, or disorders.

[0099] "Prevent" as used herein refers to a manner implemented in order to stop or delay the occurrence of a disease or a disorder or a symptom in a body.

[0100] "Effective amount" as used herein refers to an amount or dosage of the traditional Chinese medicine composition of the present application which provides a desired treatment after being administered to a subject or an organ in a single or multiple doses.

[0101] "Subject" as used herein can be a human or a non-human mammal, and can also be a cell, tissue or organ of a human or a non-human mammal, which can be a wild animal, a zoo animal, an economic animal, a pet, a laboratory animal, etc. Preferably, the non-human mammal includes but is not limited to a pig, a cow, a sheep, a horse, an ass, a fox, a raccoon dog, a mink, a camel, a dog, a cat, a rabbit, a mouse (e.g. a rat, a mouse, a guinea pig, a hamster, a gerbil, a chinchilla, a squirrel) or a monkey, etc. BRIEF DESCRIPTION OF DRAWINGS

[0102] Hereinafter, embodiments of the present application will be described in detail with reference to the accompanying drawings, in which:

[0103] Figure 1 Fig. 1 is a weight change curve of rats in each group before modeling, 4 weeks, 8 weeks, 12 weeks after modeling, 4 weeks and 8 weeks after administration, wherein NC represents a sham operation group, Model represents a model group, and AS represents an alendronate sodium tablet group;

[0104] Figure 2 Fig. 2 is a grip strength change curve of rats in each group before modeling, 4 weeks, 8 weeks, 12 weeks after modeling, 4 weeks and 8 weeks after administration, wherein NC represents a sham operation group, Model represents a model group, and AS represents an alendronate sodium tablet group;

[0105] Figure 3 Fig. 3 is a violin plot of trabecular bone thickness, trabecular bone separation, bone volume fraction and bone mineral density (BMD) of rats in each group after 8 weeks of administration, wherein NC represents a sham operation group, and Model represents a model group;

[0106] Figure 4 Fig. 4 is an HE staining diagram of the triceps surae of rats after 12 weeks of modeling, wherein Fig. A represents a sham operation group, and Fig. B represents a model group;

[0107] Figure 5 Fig. 5 is an HE staining diagram of the femur of rats after 12 weeks of modeling, wherein Fig. A represents a sham operation group, and Fig. B represents a model group;

[0108] Figure 6 Fig. 6 is an HE staining diagram of the triceps surae of rats in each group after 8 weeks of administration, wherein Fig. A represents a sham operation group, Fig. B represents a model group, Fig. C represents an alendronate sodium tablet group, and Fig. D represents a traditional Chinese medicine composition treatment group;

[0109] Figure 7: HE staining of the femurs of rats in each group after 8 weeks of drug administration, wherein Figure A represents the sham operation group, Figure B represents the model group, Figure C represents the alendronate sodium tablet group, and Figure D represents the Chinese medicine composition treatment group;

[0110] Figure 8 : Change curves of irisin content at 4, 8, and 12 weeks after modeling, where NC represents the sham operation group and Model represents the model group;

[0111] Figure 9 : Violin plots of irisin content in rats of each group after 8 weeks of drug administration, where NC: sham operation group; Model: model group;

[0112] Figure 10 : Changes in serum alkaline phosphatase (ALP) levels at 4, 8, and 12 weeks after modeling, where NC represents the sham operation group and Model represents the model group;

[0113] Figure 11 : Violin plots of serum alkaline phosphatase (ALP) levels in rats of each group at 8 weeks of drug administration, where NC: sham operation group; Model: model group;

[0114] Figure 12 : Violin plots of estradiol (E2) levels in rats of each group after 8 weeks of drug administration, where NC: sham operation group; Model: model group;

[0115] Figure 13 : Violin plots of the C-terminal peptide of type Ⅰ collagen (CTX-Ⅰ) content in rats of each group after 8 weeks of drug administration, where NC: sham operation group; Model: model group; AS: alendronate sodium tablets group;

[0116] Figure 14 : Violin plots of myostatin levels in rats of each group after 8 weeks of drug administration, where NC: sham operation group; Model: model group; AS: alendronate sodium tablets group;

[0117] Figure 15 : Violin plots of osteoprotegerin (OPG) levels in rats in each group after 8 weeks of drug administration, where NC: sham operation group; Model: model group; AS: alendronate sodium tablets group. DETAILED DESCRIPTION

[0118] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, not all. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0119] 1. Experimental control drug

[0120] Alendronate sodium tablets (Guojiazhunzi J20130085, specifications: 70mg, Hangzhou MSD Pharmaceutical Co., Ltd.).

[0121] Dexamethasone sodium phosphate injection (Sancaiqi Pharmaceutical Co., Ltd., Guojiazhunzi H44025148, batch number 20190350).

[0122] Penicillin sodium for injection (Shandong Lu Kang Pharmaceutical Co., Ltd., Guojiazhunzi H37020080, batch number 190405).

[0123] 2. Source and preparation of traditional Chinese medicine

[0124] Radix et Rhizoma Drynariae was purchased from Beijing Qiancao Traditional Chinese Medicine Decoction Pieces Co., Ltd., batch number: 240316002;

[0125] Radix Eucommiae was purchased from Beijing Qiancao Traditional Chinese Medicine Decoction Pieces Co., Ltd., batch number: 240406005;

[0126] Fructus Psoraleae was purchased from Beijing Qiancao Traditional Chinese Medicine Decoction Pieces Co., Ltd., batch number: 24080101;

[0127] Rhizoma Chuanxiong was purchased from Beijing Qiancao Traditional Chinese Medicine Decoction Pieces Co., Ltd., batch number: 24081801;

[0128] Radix Astragali was purchased from Beijing Qiancao Traditional Chinese Medicine Decoction Pieces Co., Ltd., batch number: 24070901;

[0129] Radix Codonopsis was purchased from Beijing Qiancao Traditional Chinese Medicine Decoction Pieces Co., Ltd., batch number: 240404008;

[0130] Radix et Rhizoma Bidentis was purchased from Beijing Qiancao Traditional Chinese Medicine Decoction Pieces Co., Ltd., batch number: 240507006;

[0131] Radix et Rhizoma Bidentis was purchased from Beijing Qiancao Traditional Chinese Medicine Decoction Pieces Co., Ltd., batch number: 24081208.

[0132] 3. Data statistics

[0133] # representing a significant difference between the model group and the sham operation group, * representing a significant difference between the model group and the drug administration group, * or # representing P<0.05, ** or ## representing P<0.01.

[0134] Example 1: Preparation method of traditional Chinese medicine composition

[0135] Weigh the components of the Chinese medicine composition according to their mass fractions in Table 1, add 10 times the volume of water of the total mass of the medicinal materials, decoct for 1 hour, filter the liquid while hot, add 8 times the volume of water to the residue and continue to decoct for 1 hour, combine the two filtrates, and concentrate to a 3 g / mL liquid.

[0136] Table 1: Mass fractions of each component in the Chinese medicine composition

[0137]

[0138]

[0139] Example 2: Modeling

[0140] Six-month-old female Sprague-Dawley rats were used. Both ovaries were removed via a dorsal approach in the model group, while equal volumes of adipose tissue surrounding the ovaries were removed in the sham group. Muscle and skin were then sutured layer by layer. Penicillin sodium (80,000 units / rat) was administered intramuscularly once daily for three days to prevent infection. One week after recovery, the model group received intraperitoneal injections of dexamethasone sodium phosphate (1 mg / kg / day) for two consecutive weeks.

[0141] At 4, 8, and 12 weeks after surgery, the rats' weight and grip strength were measured, the triceps surae and femur were stained, the serum irisin and ALP levels were measured, and bone density was measured using a dual-energy X-ray absorptiometry to verify the success of the model. The specific testing methods are as follows:

[0142] (1) Grip force detection

[0143] After 4 and 8 weeks of drug intervention, the rats' forelimb grip strength was measured using a YLS-13A rat and mouse grip strength tester. Before testing, the rats were allowed to acclimate to the grip strength tester for 10 minutes. After acclimatization, the rats' tails were grasped and gently pulled backward at a constant speed, causing the rats to loosen their grips. The grip strength reading was recorded. This procedure was repeated 10 times, and the average value was calculated and compared to body weight.

[0144] (2) Hematoxylin-eosin (HE) staining

[0145] After 8 weeks of drug intervention, rats were anesthetized and sacrificed. The left femur and gastrocnemius muscle were fixed in 4% paraformaldehyde for at least 24 hours. Paraffin sections were deparaffinized with environmentally friendly dewaxing solution for 40 minutes, hydrated with graded ethanol, and then stained with hematoxylin-eosin. High-definition constant staining was performed for 1 minute, followed by hematoxylin staining for 3-5 minutes, followed by blueing, dehydration with 95% ethanol for 1 minute, rapid staining with eosin for 15 seconds, and mounting with neutral gum after gradient clearing. Frozen sections were rewarmed at 4°C, fixed with 4% paraformaldehyde for 15 minutes, and rinsed with running water. The same staining and mounting procedures were followed, and images were acquired through microscopy.

[0146] (3) Serum biochemical index detection

[0147] After 8 weeks of drug intervention, the rats were anesthetized and blood was taken from the abdominal aorta at 4°C, 3000r·min -1 Centrifugation for 15 min, take serum, using ELISA method to detect the content of serum Irisin and serum alkaline phosphatase (ALP), the specific operation steps are strictly according to the kit instructions.

[0148] Test results:

[0149] (1) The results of rat weight are shown in Table 2.

[0150] Table 2: Rat weight

[0151] Group / time Before modeling 4 weeks after modeling 8 weeks after modeling 12 weeks after modeling Sham group 347.96±26.61 350.22±41.30 351.70±56.05 383.23±64.90 Model group 348.32±28.25 282.08 ± 29.68 ## ]] 367.57±30.62 412.32±41.87

[0152] The body weight of the model group rats decreased significantly at 4 weeks after modeling, and the body weight showed an upward trend at 8 and 12 weeks after modeling Figure 1 ).

[0153] (2) The results of grip strength are shown in Table 3.

[0154] Table 3: Grip strength

[0155] Group / time 4 weeks after modeling 8 weeks after modeling 12 weeks after modeling Sham group 3.82±0.39 3.76±0.64 4.03±0.47 Model group 3.63±0.63 3.58±0.43 3.56 ± 0.41 ## ]]

[0156] The grip strength of the model group rats showed a downward trend at 4, 8 and 12 weeks after modeling, and the grip strength decreased significantly at 12 weeks after modeling Figure 2 ).

[0157] (3) The gastrocnemius muscle of the rat calf

[0158] At 12 weeks after modeling, the HE staining results are shown in Figure 4 Among them, the intercellular arrangement of the gastrocnemius muscle of the model group rats was loose, the interstitial space increased, the connective tissue between muscle fibers proliferated, the number of muscle nuclei decreased, and the muscle nuclei were obviously shrunk, located at the edge of the cell membrane, and the central nucleus was not obvious, indicating that the muscle cells were aging.

[0159] (4) The femur of the rat

[0160] At 12 weeks after modeling, the HE staining results of the model group rats are shown in Figure 5 Among them, the trabecular bone of the model group rats decreased significantly, the connection was incomplete, the bone marrow space area increased significantly, and the fat cell pits increased significantly, indicating that the osteoporosis model was successful.

[0161] 5, Irisin

[0162] At 4 weeks after modeling, the content of serum Irisin in the model group rats showed no obvious change trend; at 8 and 12 weeks after modeling, the content of serum Irisin decreased significantly (see Table 4 and Figure 8 ). It is proved that the muscle-osteoporosis rat model is successfully established.

[0163] Table 4: Irisin

[0164]

[0165] 6、ALP

[0166] The ALP content in serum of the model group rats was significantly reduced at 4, 8 and 12 weeks after modeling (see Table 5 and Figure 10 ).

[0167] Table 5: ALP

[0168]

[0169] In summary, the rats with bilateral ovariectomy can be used as animal models of osteoporosis and sarcopenia-osteoporosis. In addition, the estrogen of the model group is significantly lower than that of the sham operation group, which proves that it can be used as an animal model of menopausal osteoporosis.

[0170] Example 3: Treatment of osteoporosis and sarcopenia-osteoporosis

[0171] 1. Grouping and dosing regimen

[0172] After the rats were adaptively fed for 1 week, except for the sham operation group (Sham), they were randomly divided into the model group (Model), the positive drug alendronate sodium tablet group (AS), and the prescription 3 group according to the body weight, with 10 rats in each group. The modeling method was the same as that in Example 2. After 12 weeks of modeling, drug intervention was started. The dosage of prescription 3 was 7.6125 g / kg. Alendronate sodium tablets were administered by gavage at 7.35 mg / kg per day. The Sham group was given pure water for gavage every day. After 8 weeks of continuous gavage, the samples were taken, the body weight, grip strength, rat calf triceps, and femur staining (the method is referred to Example 2) were performed, and the rat femur was detected by Micro-CT. The specific steps of the Micro-CT detection method are as follows: after 8 weeks of administration, the rats were anesthetized and sacrificed, the right femur of the rats was completely separated, the soft tissues such as muscles on the femur were removed, and 4% paraformaldehyde was used for fixation. Micro-CT scanning was performed. The target region ROI was analyzed using data analysis software, all samples were analyzed in the same region, the required parameter values were obtained, and the data were exported. The original image was reconstructed using three-dimensional reconstruction software Recon.

[0173] 2. Results

[0174] 1) The results of the body weight of rats are shown in Figure 1 and Table 6.

[0175] Table 6: Body weight of rats

[0176] Group / time 4 weeks after administration 8 weeks after administration Sham group 374.55±49.66 373.99±52.94 Model group 469.07 ± 50.65 ## ]] 501.43 ± 55.28 ## ]] Positive drug group 424.73 ± 58.93 * ]] 443.49 ± 61.37 * ]] Prescription 3 400.98 ± 35.16 ** ]] 424.07 ± 45.14 ** ]]>

[0177] The body weight of the model group rats was significantly increased at 4 and 8 weeks of administration, the body weight of the positive drug (alendronate sodium) group and the prescription 3 group rats was significantly decreased after administration, and the body weight of the prescription 3 group was decreased more significantly than that of the positive drug alendronate sodium group.

[0178] 2) The grip strength test results are shown in Table 6. Figure 2 and Table 7.

[0179] Table 7: Grip strength

[0180] Group / time 4 weeks after administration 8 weeks after administration Sham group 4.16±0.54 5.01±0.72 Model group 3.58 ± 0.42 ## ]] 3.64 ± 0.35 ## ]] Positive drug group 4.03 ± 0.62 * ]] 4.16 ± 0.56 * ]] Prescription 3 <![CDATA[4.31±0.44 ** ]]> 4.05 ± 0.35 * ]]

[0181] The grip strength of the model group rats was significantly decreased at 4 and 8 weeks of administration, the grip strength of the alendronate sodium group and the prescription 3 group rats was significantly increased after administration, and the grip strength of the prescription 3 group was increased more significantly than that of the positive drug alendronate sodium group at 4 weeks.

[0182] 3) Femur Micro-CT detection

[0183] The femur Micro-CT detection results at 8 weeks of administration are shown in Table 8. Figure 3 and Table 8.

[0184] Table 8: Femur Micro-CT detection

[0185]

[0186]

[0187] The trabecular bone thickness, bone volume fraction and bone mineral density of the model group rats were significantly decreased, and the trabecular bone separation degree was significantly increased at 8 weeks of administration. The trabecular bone thickness, bone volume fraction and bone mineral density of the alendronate sodium group and the prescription 3 group were significantly increased, and the trabecular bone separation degree was significantly decreased.

[0188] 4) Rat calf triceps

[0189] At 8 weeks of administration, the intercellular arrangement of the calf triceps of the model group rats was loose, the gap was increased, the connective tissue between muscle fibers was proliferated, the number of muscle nuclei was reduced, the muscle was obviously shrunk, the muscle nuclei were located at the edge of the cell membrane, and the central nuclei were not obvious. The intercellular arrangement of the calf triceps of the positive drug and prescription 3 group rats was relatively neat, the gap was reduced, the muscle connective tissue proliferation was not obvious, and the number of muscle nuclei was increased. Figure 6

[0190] 5) Rat femur

[0191] ​The pathological results of the model group rats showed that the bone tissue of the sham operation group rats was more continuous, the morphology was complete, and there was no obvious pathological change. The bone tissue of the model group rats was thin and narrow, not continuous, the free end increased, the bone marrow cavity widened, a large number of osteoclasts and vacuolated fat cells appeared, the bone cell and bone volume decreased; the bone tissue structure of the alendronate sodium tablet and prescription 3 group rats was relatively complete, the bone trabecula was slightly more, the bone plate layer was denser, the bone marrow cavity was reduced, the fat vacuoles were obviously reduced, and the bone cells were more Figure 7

[0192] Example 4: Biochemical index detection proves the treatment of osteoporosis and sarcopenia-osteoporosis

[0193] After the rats were adaptively fed for 1 week, except the sham operation group (Sham), they were randomly divided into a model group (Model), and a prescription 3 group according to the body weight, 10 rats in each group. The modeling method was the same as that in Example 2, and after modeling for 12 weeks, drug intervention was started.

[0194] Among them, the prescription 3 group was given a dose of 30.45 g / kg, the Sham group was given pure water for gavage every day, and the sample was taken after continuous gavage for 8 weeks. The contents of iridoid and ALP in serum were detected by ELISA method (the method was the same as that in Example 2).

[0195] 1, the results of iridoid detection are shown in Table 9 and Figure 9 .

[0196] Table 9: Iridoid

[0197] Group / time 8 weeks after administration (pg / ml) Sham group 691.17±273.39 Model group 328.51 ± 240.90 ## ]] Prescription 3 496.60 ± 138.83 * ]]

[0198] The serum iridoid of the model group rats was significantly reduced after 8 weeks of administration, and the serum iridoid of the prescription 3 group rats was significantly increased after administration. The content of iridoid was increased after administration, which played a role in protecting the skeleton.

[0199] 2, the results of ALP are shown in Figure 11 and Table 10.

[0200] Table 10: ALP

[0201] Group / time 8 weeks after administration (ng / ml) Sham group 2.15±0.96 Model group 5.86 ± 3.68 ## ]] Prescription 3 3.57 ± 0.94 * ]]

[0202] The ALP content in the serum of the model group rats was significantly increased after 8 weeks of administration, and the ALP content in the serum of the prescription 3 group rats was significantly reduced after administration, indicating that the bone turnover rate was reduced.

[0203] Example 5: Treatment of menopausal osteoporosis

[0204] ​After the rats were adaptively fed for 1 week, except the sham operation group (Sham), they were randomly divided into a model group (Model), and a prescription 3 group, 10 rats in each group. The modeling method was the same as in Example 2. After modeling for 12 weeks, drug intervention was started.

[0205] The dosage of the prescription 3 group was 15.225 g / kg, and the Sham group was given pure water for gavage every day. The samples were taken after continuous gavage for 8 weeks. The change of estrogen in serum was detected by the ELISA method in Example 2.

[0206] The results of the change of estrogen at the time of administration for 8 weeks are shown in Table 11. Figure 12 and Table 11.

[0207] Table 11: Estrogen

[0208]

[0209]

[0210] The E2 content in the serum of the model group rats was significantly reduced at the time of administration for 8 weeks, and the E2 content in the serum of the prescription 3 group rats was significantly increased after administration. It shows that the traditional Chinese medicine composition of the application has the effect of treating menopausal osteoporosis.

[0211] Example 6: Results of serum marker detection

[0212] After the rats were adaptively fed for 1 week, except the sham operation group (Sham), they were randomly divided into a model group (Model), a positive drug alendronate sodium tablet group (AS), and a prescription 3 group, 10 rats in each group. The modeling method was the same as in Example 2. After modeling for 12 weeks, drug intervention was started. The dosage of the prescription 3 was 30.45 g / kg. The alendronate sodium tablet was gavaged at 7.35 mg / kg per day, and the Sham group was given pure water for gavage every day. The samples were taken after continuous gavage for 8 weeks. The serum markers were detected by the ELISA method in Example 2, and the results are shown in Table 12 and Figures 13-15 .

[0213]

[0214] The increase of CTX-Ⅰ (type Ⅰ collagen C-terminal peptide) is usually related to osteoporosis and increased bone resorption. CTX-Ⅰ is an important bone metabolism index reflecting bone resorption, and its level increase means increased bone resorption, which is related to osteoporosis. The CTX-Ⅰ content in the serum of the model group rats was significantly increased at the time of administration for 8 weeks, and the CTX-Ⅰ content in the serum of the rats was significantly reduced after administration, that is, the bone resorption was reduced, which played a therapeutic role. Figure 13

[0215] ​Myostatin is a negative regulator of muscle growth and regeneration, which is secreted by muscle cells. Myostatin inhibits the differentiation of osteoblasts and stimulates the activity of osteoclasts in a dose-dependent manner. The content of Myostatin in the serum of the model group rats increases after 8 weeks of administration, and the content of Myostatin in the serum of the administration group shows a downward trend, which has a significant effect on inhibiting the differentiation of osteoblasts Figure 14 )。

[0216] The main functions of OPG (osteoprotegerin) include inhibiting osteoclastogenesis, promoting vascular maturation, inhibiting tumor cell apoptosis, and playing a role in various diseases. The content of OPG in the serum of the model group rats significantly increases after 8 weeks of administration, and the content of OPG in the serum shows a downward trend after administration, and the content of OPG in the serum significantly decreases, indicating that osteoclastogenesis is successfully inhibited, and the purpose of treating osteoporosis is achieved Figure 15 )。

[0217] The above describes the preferred embodiments of the present application, but the present application is not limited to the specific details in the above embodiments, and various simple modifications can be made to the technical solutions of the present application within the technical concept of the present application, and these simple modifications all belong to the protection scope of the present application.

[0218] In addition, it should be noted that each specific technical feature described in the above specific embodiments can be combined in any appropriate manner without contradiction, and in order to avoid unnecessary repetition, the present application will not further describe various possible combinations.

Claims

1. A Chinese medicine composition, characterized in that The traditional Chinese medicine composition comprises rhizoma drynariae, eucommia bark, psoralea corylifolia, ligusticum chuanxiong, raw astragalus, codonopsis pilosula, cyathula and rhizoma schizonepetae; rhizoma drynariae, eucommia bark and psoralea corylifolia are the main drugs.

2. The Chinese medicine composition according to claim 1, characterized in that The traditional Chinese medicine composition comprises, by mass, 8-20 parts of drynaria rhizome, 10-20 parts of eucommia bark, 5-15 parts of psoralea corylifolia, 10-20 parts of chuanxiong, 20-40 parts of raw astragalus, 20-40 parts of codonopsis pilosula, 10-20 parts of cyathula and 10-20 parts of radix polygoni multiflori.

3. The Chinese medicine composition according to claim 1 or 2, characterized in that The Chinese medicinal ingredients in the Chinese medicine composition are, by weight, 8-20 parts of Drynaria fortunei, 10-20 parts of Eucommia ulmoides, 5-15 parts of Psoralea corylifolia, 10-20 parts of Chuanxiong, 20-40 parts of Raw Astragalus, 20-40 parts of Codonopsis pilosula, 10-20 parts of Cyathula capitata and 10-20 parts of Polygonum multiflorum, among which the monarch drugs are Drynaria fortunei, Eucommia ulmoides and Psoralea corylifolia.

4. The Chinese medicine composition according to any one of claims 1 to 3, characterized in that The traditional Chinese medicine composition comprises, by weight, 15 parts of drynaria rhizome, 15 parts of eucommia bark, 10 parts of psoralea corylifolia, 15 parts of ligusticum chuanxiong, 30 parts of raw astragalus, 30 parts of codonopsis pilosula, 15 parts of cyathula and 15 parts of rhizoma achyranthis Bidentatae.

5. The Chinese medicine composition according to any one of claims 1 to 4, characterized in that The Eucommia ulmoides is salt Eucommia ulmoides; the Psoralea corylifolia is salt Psoralea corylifolia; the Drynaria rhizome is scalded Drynaria rhizome; Preferably, the salted Eucommia ulmoides is prepared by frying the Eucommia ulmoides with salt water and breaking the silk into pieces; Preferably, the salted psoralea corylifolia is prepared by mixing psoralea corylifolia with salt water, slightly moistening it, and then frying it in a pot over a low heat until it is slightly swollen. Preferably, the scalded Drynaria Rhizoma is prepared by sand scalding method.

6. The Chinese medicine composition according to any one of claims 1 to 5, characterized in that The dosage forms of the Chinese medicine composition include tablets, capsules, powders, pills, granules, injections, decoctions, wine preparations, ointments, pastes or oral liquids.

7. The Chinese medicine composition according to any one of claims 1 to 6, characterized in that The traditional Chinese medicine composition further includes pharmaceutically acceptable excipients; preferably, the excipients include one or more of a diluent, a disintegrant, a lubricant, an excipient, a binder, a glidant, a filler or a surfactant.

8. A method for preparing the traditional Chinese medicine composition according to any one of claims 1 to 7, characterized in that: The preparation method includes powdering, decocting, frying, soaking, percolation, supercritical extraction or reflux extraction; Decoction is preferred.

9. Use of the traditional Chinese medicine composition according to any one of claims 1 to 7 or the traditional Chinese medicine composition obtained by the preparation method according to claim 8 in preparing a medicament for treating a disease, characterized in that: The disease is osteoporosis; Preferably, the disease is sarcopenia-osteoporosis or menopausal osteoporosis.

10. A method for improving muscle structure and bone tissue structure, characterized in that: The method comprises administering the traditional Chinese medicine composition according to any one of claims 1 to 7 or the traditional Chinese medicine composition obtained by the preparation method according to claim 8.

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