Pharmaceutical composition for treating osteoporosis and preparation and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a pharmaceutical composition for treating osteoporosis and a preparation and application thereof. The pharmaceutical composition is prepared from icariin, echinacoside, leonurus glycoside and cordycepin as raw materials according to a certain weight percentage. The traditional Chinese medicine composition can be prepared into various clinical common oral preparations. Pharmacological experiments show that the pharmaceutical composition can significantly inhibit the formation of osteoclasts in vitro. In addition, the pharmaceutical composition disclosed by the invention can be used for remarkably improving the bone mineral density value and the number of bone trabecula of ovariectomized mice. Experimental results also show that after icariin, echinacoside, leonurus glycoside and cordycepin are combined for use, a remarkable synergistic interaction effect is achieved. The pharmaceutical composition disclosed by the invention has a prospect of being developed into a medicine for treating osteoporosis, and has important social benefits and economic benefits.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a pharmaceutical composition for treating osteoporosis, a preparation thereof and a use thereof. Background Art

[0002] Osteoporosis (OP) is a systemic bone disease characterized by decreased bone mass, damaged bone microstructure, increased bone fragility, and susceptibility to fractures. Osteoporosis is divided into two categories: primary and secondary. Primary osteoporosis includes postmenopausal osteoporosis (type I), senile osteoporosis (type II), and idiopathic osteoporosis (including adolescent type). Primary osteoporosis is more common in postmenopausal women and elderly men. There are usually no obvious clinical manifestations in the early stage. As the disease progresses, bone mass continues to be lost, and patients will experience pain and spinal deformity. In severe cases, osteoporotic fractures may occur. Some patients experience decreased muscle capacity and strength, and are prone to falling, which increases the risk of fractures and reduces the quality of life. Osteoporotic fractures are extremely harmful and are one of the main causes of disability and death in elderly patients. The etiology and pathogenesis of senile osteoporosis are multifaceted, and organ dysfunction caused by aging is the main factor. In addition to endocrine factors, a variety of cytokines also affect bone metabolism and reduce osteogenic activity. For example, insufficient intake of calcium and vitamin D will lead to insufficient conversion of provitamin D to vitamin D in the skin, decreased renal function, and insufficient hydroxylation of vitamin D. The osteogenic differentiation ability of bone marrow mesenchymal stem cells decreases. Muscle atrophy, reduced stress stimulation to bones, and impaired regulation of bone metabolism. All these factors affect bone metabolism, resulting in insufficient bone formation, excessive bone resorption, bone loss, damage to bone structure, and osteoporosis.

[0003] At present, the research on drugs for the treatment of osteoporosis is mainly based on two strategies. One is to develop drugs for the purpose of increasing osteoblast differentiation and promoting bone formation, aiming to increase bone mass by increasing osteoblasts and osteocytes to supplement the missing bone tissue; the other is to develop drugs for the purpose of promoting osteoclast apoptosis and inhibiting osteoclast activation, aiming to reduce bone resorption, alleviate the continuous loss of bone mass, and improve osteoporosis and related symptoms. At present, the clinical treatment of osteoporosis is achieved through the above two mechanisms. The main commercially available drugs are calcium preparations, bisphosphonates, estrogens, androgens, vitamin D3, etc. However, these drugs have many side effects when used for a long time. For example, long-term use of bisphosphonates has toxic side effects on tissues and organs such as the kidneys, liver and gastrointestinal tract; long-term use of parathyroid hormone drugs will increase the risk of osteosarcoma. Therefore, it is very necessary and urgent to develop medical products and compositions for the prevention, improvement and treatment of osteoporosis with high safety, low side effects and suitable for long-term use.

[0004] Traditional Chinese medicine in my country has unique advantages in regulating chronic and complex diseases. Osteoporosis belongs to the category of "bone atrophy" and "bone arthritis" in traditional Chinese medicine. Traditional Chinese medicine believes that "kidneys govern bones, produce marrow and store essence", which means that the growth and development of human bones depend on kidney essence. Especially after women's menopause or elderly men enter menopause, kidney qi declines from prosperity, and kidney essence deficiency often occurs. Kidney essence deficiency leads to emptiness of bone marrow, which cannot nourish bones, and bones will be lost, causing osteoporosis. In addition, with the increase of age, it is often accompanied by the occurrence of blood stasis, which causes bone nutrition disorders and aggravates the occurrence of osteoporosis. The treatment of osteoporosis should be based on tonifying the kidney and replenishing qi, activating blood circulation and strengthening bones. Many Chinese medicine prescriptions are used for the treatment of osteoporosis based on this method, but they generally have shortcomings such as inconvenience in carrying and taking, and unclear ingredients. At the same time, due to the large number of Chinese medicinal flavors in Chinese medicine prescriptions, the cost and safety risks are high, which is not conducive to effective quality control and large-scale production.

[0005] In view of this, it is imperative to use modern pharmacological research methods to conduct in-depth research and screening of natural product active ingredient monomers in classic Chinese medicine prescriptions, and then develop a pharmaceutical composition for the treatment of osteoporosis with good efficacy, few side effects and easy use. Summary of the invention

[0006] The purpose of the present invention is to provide a safe, effective, convenient and economical pharmaceutical composition for osteoporosis patients, and to provide a new idea for the clinical treatment of osteoporosis and the modernization of traditional Chinese medicine.

[0007] Specifically, the present invention is realized through the following technical solutions:

[0008] In a first aspect, the present invention provides a pharmaceutical composition for treating osteoporosis, the pharmaceutical composition comprising icariin, echinacoside, leonuriside and cordycepin.

[0009] As an optional mode, in the above pharmaceutical composition, the pharmaceutical composition comprises the following active ingredients by weight percentage: 20%-50% icariin, 20%-50% echinacoside, 10%-30% leonuri glycoside and 5%-20% cordycepin.

[0010] As an optional mode, in the above-mentioned pharmaceutical composition, the pharmaceutical composition comprises the following active ingredients by weight percentage: 30%-40% icariin, 30%-40% echinacoside, 10%-20% leonuri glycoside and 7%-15% cordycepin.

[0011] As an optional manner, in the above pharmaceutical composition, the pharmaceutical composition comprises the following active ingredients by weight percentage: 40% icariin, 30% echinacoside, 20% leonuriside and 10% cordycepin.

[0012] As an optional method, in the above-mentioned pharmaceutical composition, the four natural product monomer components of icariin, echinacoside, leonuri and cordycepin are used in combination, which has a significant synergistic effect and can significantly enhance the effect of the pharmaceutical composition of the present invention in treating osteoporosis.

[0013] Those skilled in the art can anticipate that the four natural product monomer components of icariin, echinacoside, leonuri and cordycepin, the active ingredients used in the pharmaceutical composition of the present invention, can be extracted and separated from traditional Chinese medicinal materials containing the above-mentioned active ingredients, such as epimedium, cistanche deserticola, rehmannia root, cordyceps and the like, by a biological purification method, or can be directly purchased from commercially available products.

[0014] In a second aspect, the present invention provides a pharmaceutical preparation for treating osteoporosis, wherein the pharmaceutical preparation comprises the pharmaceutical composition described in the first aspect, and a pharmaceutically acceptable carrier or excipient.

[0015] As an optional mode, in the above-mentioned pharmaceutical preparation, the pharmaceutical preparation is an oral dosage form.

[0016] As an optional mode, in the above-mentioned pharmaceutical preparation, the oral dosage form is capsule, tablet, granule or oral liquid.

[0017] In addition, those skilled in the art can anticipate that the four natural product monomer components of icariin, echinacoside, leonuri and cordycepin used in the pharmaceutical composition of the present invention can also be administered sequentially, concomitantly or simultaneously in different pharmaceutical preparations.

[0018] In a third aspect, the present invention provides use of the pharmaceutical composition described in the first aspect or the pharmaceutical preparation described in the second aspect in the preparation of a drug for treating osteoporosis.

[0019] As an optional mode, in the above use, the osteoporosis is primary osteoporosis or secondary osteoporosis.

[0020] As an optional mode, in the above-mentioned use, the primary osteoporosis is postmenopausal osteoporosis (type I), senile osteoporosis (type II) or idiopathic osteoporosis (including juvenile type).

[0021] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] (1) Based on the classical prescriptions and commonly used Chinese medicinal materials for the treatment of osteoporosis in clinic, the present invention selects four natural product monomers through a large number of pharmacological experiments and screens them, and combines them in a specific proportion to prepare a pharmaceutical composition capable of treating osteoporosis, which provides a new idea for the modernization of traditional Chinese medicine.

[0024] (2) Pharmacological experiments have shown that the pharmaceutical composition of the present invention can significantly inhibit the formation of osteoclasts in vitro. When the weight ratio of icariin, echinacoside, leonurin and cordycepin in the pharmaceutical composition is 4:3:2:1, the effect is best, and the inhibition rate of osteoclasts can be as high as 84.78%. In addition, the pharmaceutical composition of the present invention can significantly improve the bone density value and the number of trabeculae in ovariectomized mice. When the weight ratio of icariin, echinacoside, leonurin and cordycepin in the pharmaceutical composition is 4:3:2:1, the effect is best. The experimental results also show that the combination of icariin, echinacoside, leonurin and cordycepin has a significant synergistic effect, has no toxic side effects, and has a good clinical application prospect.

[0025] (3) The various natural product monomers used in the pharmaceutical composition of the present invention have a wide range of sources and can be produced by mature methods, making the pharmaceutical composition of the present invention convenient for industrial large-scale production. In addition, by adding a pharmaceutically acceptable carrier, the pharmaceutical composition of the present invention is made into a conventional oral preparation, which makes the drug treatment stable, improves the efficacy of the drug, and is easy to take. DETAILED DESCRIPTION

[0026] The inventors conducted in-depth research on the pharmacological mechanisms of the commonly used classical prescriptions for treating osteoporosis and the commonly used Chinese medicinal materials for treating osteoporosis, and found for the first time through extensive screening that the combined use of natural product monomers icariin, echinacoside, leonurin and cordycepin can significantly enhance the effect of treating osteoporosis. The present invention was completed on this basis.

[0027] The natural product monomers used in the pharmaceutical composition of the present invention are as follows:

[0028] The chemical structure of Icraiin is Icariin has a wide range of pharmacological effects, including immunomodulation, anti-oxidation, inhibition of cell apoptosis and angiogenesis, etc. It has a significant effect on inhibiting congestive heart failure, alleviating myocardial ischemia-reperfusion injury, inhibiting myocardial cell apoptosis, and improving the progression of atherosclerosis and myocardial infarction.

[0029] The chemical structure of "Echinacoside" is Echinacoside is a phenylethanol glycoside compound extracted from traditional Chinese medicine such as Cistanche deserticola, which has significant pharmacological activities such as anti-inflammatory, antioxidant, and neuroprotective.

[0030] The chemical structure of "Ajugol" is Leonurusin is an iridoid glycoside.

[0031] "Cordycepin" is 3'-deoxyadenosine, and its chemical structure is Cordycepin is a nucleoside substance derived from Cordyceps militaris. It has a wide range of pharmacological activities and has been widely studied in the anti-tumor, anti-leukemia and immunomodulatory aspects. It also has cardiovascular protective effects such as anti-aging, anti-inflammatory, hypoglycemic and hypolipidemic effects.

[0032] As used herein, "pharmaceutically acceptable carrier" refers to a conventional drug carrier in the field of pharmaceutical preparations, selected from one or more of fillers, binders, disintegrants, lubricants, suspending agents, wetting agents, pigments, flavoring agents, solvents, and surfactants.

[0033] The filler of the present invention includes but is not limited to starch, microcrystalline cellulose, sucrose, dextrin, lactose, powdered sugar, glucose, etc.; the lubricant includes but is not limited to magnesium stearate, stearic acid, sodium chloride, sodium oleate, sodium lauryl sulfate, poloxamer, etc.; the binder includes but is not limited to water, ethanol, starch slurry, syrup, hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, sodium alginate, polyvinyl pyrrolidone, etc.; the disintegrant includes but is not limited to starch effervescent mixture, i.e. sodium bicarbonate and citric acid, tartaric acid, low-substituted hydroxypropyl cellulose, etc.; the suspending agent includes but is not limited to polysaccharides such as acacia gum, agar, alginic acid, cellulose ether and carboxymethyl chitosan, etc.; the solvent includes but is not limited to water, a balanced salt solution, etc.

[0034] The pharmaceutical composition can be prepared into various solid oral preparations, liquid oral preparations, etc. Pharmaceutically acceptable oral solid preparations include: ordinary tablets, dispersible tablets, enteric-coated tablets, granules, capsules, dripping pills, powders, etc., and oral liquid preparations include oral liquids, emulsions, etc.

[0035] The above-mentioned various dosage forms can be prepared according to conventional processes in the field of pharmaceutical preparations.

[0036] In the medical uses described above, the administration time, number of administrations, and frequency of administration of the composition of the present invention need to be determined according to the specific diagnosis results of the disease, which is within the technical scope mastered by those skilled in the art.

[0037] The treatment plan for mice can be applied to humans, and the effective doses of all drugs for humans can be converted by the effective doses of the drugs for mice, which is also easy to achieve for ordinary technicians in this field.

[0038] The present invention will be further described with reference to specific embodiments below. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the scope of the present invention.

[0039] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0040] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are commercially available products unless otherwise specified.

[0041] Unless otherwise specified, all percentages and parts in the present invention are percentages and parts by weight.

[0042] Example 1: Effect of the pharmaceutical composition of the present invention on osteoclast activity

[0043] Excessive proliferation and activation of osteoclasts can cause increased bone resorption, reduce bone mass, reduce bone density, increase bone fragility and fracture risk. Therefore, inhibiting the differentiation of bone marrow mononuclear macrophages into osteoclasts helps prevent and treat osteoporosis. Bone marrow mononuclear cells were isolated from the femur and tibia of 1-month-old mice (purchased from Liaoning Changsheng Biotechnology Co., Ltd.) and evenly plated in 24-well plates, with 1×10 per well. 5 After the cells adhered to the wall, they were cultured with α-MEM medium containing 25ng / mL M-CSF, 25ng / mL RANKL and 10% fetal bovine serum. At the same time, except for the normal control group, the other groups added different drug compositions, and the culture medium was replaced every 3 days. After 6 days, TRAP staining was used to calculate the osteoclast data in each group. The number of osteoclast positive cells was defined as: TRAP positive cells with more than 3 nuclei.

[0044] The experimental data are expressed as mean ± SD, and the data statistics are analyzed by one-way ANOVA using SPSS25.0. Compared with the normal group, * P<0.05, ** P<0.01, *** P<0.001.

[0045] The experimental results are shown in Table 1.

[0046] Table 1: Number of osteoclasts and differentiation inhibition rate

[0047]

[0048]

[0049] The experimental results in Table 1 show that the pharmaceutical composition of the present invention can significantly inhibit the formation of osteoclasts in vitro. Among them, when the weight ratio of icariin, echinacoside, leonuri and cordycepin in the pharmaceutical composition is 4:3:2:1, the best effect is achieved, and the inhibition rate of osteoclasts can be as high as 84.78%.

[0050] In contrast, it can be seen from the results of each control group that when the types of components in the pharmaceutical composition are changed and replaced with components with similar efficacy, or the dosage ratio of the components in the pharmaceutical composition is changed, or some components are removed from the pharmaceutical composition, the effect of the pharmaceutical composition in inhibiting osteoclast formation is significantly reduced.

[0051] The above results indirectly illustrate that the effect of the pharmaceutical composition of the present invention in inhibiting osteoclast activation is not due to one or several components, but is the result of the synergistic and joint action of various components.

[0052] Example 2: Effect of the pharmaceutical composition of the present invention on bone density and trabecular bone in ovariectomized mice

[0053] The female C57BL / 6 mice used in this embodiment were purchased from Liaoning Changsheng Biotechnology Co., Ltd. The mice were kept at a temperature of 23±2°C, a humidity of 55±5%, a ventilation frequency of 12-15 times / hour, a lighting time of 12 hours / day (7:00-19:00 light), and 5 mice / cage (235×325×170H mm). The breeding conditions were solid feed (purchased from Liaoning Changsheng Biotechnology Co., Ltd.) and free diet. When the mice were 3 months old, they were randomly divided into a sham operation group, a model group, and a drug-treated group. The mice were fasted but not watered 12 hours before surgery. The mice were anesthetized with isoflurane, and the skin was then cleaned and disinfected with 75% ethanol. Each group of mice underwent surgery through a ventral incision. Except for the sham operation group mice, which did not remove the ovaries, the other groups of mice underwent bilateral ovariectomy, and all surgical mice were intraperitoneally injected with penicillin solution. After the operation, the operated animals were placed in an incubator and transferred to a clean cage for recovery for 1 week after they woke up. During this period, penicillin solution was injected intraperitoneally every other day to prevent wound infection in mice.

[0054] Except for the mice in the sham operation group and the model group, the mice in each drug administration group were gavaged with the corresponding dose of drugs (see Table 2) once a day for 2 consecutive months. Twelve hours after the last administration, the mice were anesthetized and the bone mineral density (BMD) and the number of trabeculae in the right hind limbs of the mice were scanned.

[0055] The data are expressed as mean ± SD, and the data were analyzed using one-way ANOVA of SPSS25.0. Compared with the sham operation group, *** P<0.001; compared with the model group, #P<0.05, ## P<0.01, ### P<0.001.

[0056] The experimental results are shown in Table 2.

[0057] Table 2: BMD and trabecular number in ovariectomized mice

[0058]

[0059]

[0060] The experimental results in Table 2 show that the bone density and the number of trabeculae in the model group mice were significantly reduced compared with those in the sham operation group mice. The pharmaceutical composition of the present invention can significantly improve the bone density value and the number of trabeculae in ovariectomized mice. Among them, the best effect is achieved when the weight ratio of icariin, echinacoside, leonuri and cordycepin in the pharmaceutical composition is 4:3:2:1.

[0061] In contrast, it can be seen from the results of each control group that when the types of components in the pharmaceutical composition are changed and replaced with components with similar efficacy, or the dosage ratio of the components in the pharmaceutical composition is changed, or some components are removed from the pharmaceutical composition, the improvement effect of the pharmaceutical composition on the bone density value and the number of trabeculae in ovariectomized mice is significantly reduced.

[0062] The above results indirectly illustrate that the improvement of the bone mineral density and trabecular number of ovariectomized mice by the pharmaceutical composition of the present invention is not caused by one or several components, but is the result of the synergistic and joint action of the components.

[0063] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A pharmaceutical composition for treating osteoporosis, characterized in that: The pharmaceutical composition comprises icariin, echinacoside, leonuriside and cordycepin.

2. The pharmaceutical composition according to claim 1, characterized in that: The pharmaceutical composition comprises the following active ingredients by weight: 20%-50% icariin, 20%-50% echinacoside, 10%-30% leonuri glycoside and 5%-20% cordycepin.

3. The pharmaceutical composition according to claim 1 or claim 2, characterized in that: The pharmaceutical composition comprises the following active ingredients by weight: 30%-40% icariin, 30%-40% echinacoside, 10%-20% leonuriside and 7%-15% cordycepin.

4. A pharmaceutical preparation for treating osteoporosis, characterized in that: The pharmaceutical preparation comprises the pharmaceutical composition according to any one of claims 1 to 3, and a pharmaceutically acceptable carrier or excipient.

5. The pharmaceutical preparation according to claim 4, characterized in that: The pharmaceutical preparation is an oral dosage form.

6. The pharmaceutical preparation according to claim 5, characterized in that: The oral dosage form is capsule, tablet, granule or oral liquid.

7. Use of the pharmaceutical composition according to any one of claims 1 to 3 or the pharmaceutical preparation according to any one of claims 4 to 6 in the preparation of a medicament for treating osteoporosis.

8. The use according to claim 7, characterized in that: The osteoporosis is primary osteoporosis or secondary osteoporosis.

9. The use according to claim 8, characterized in that: The primary osteoporosis is postmenopausal osteoporosis, senile osteoporosis or idiopathic osteoporosis.

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