阅读说明：本技术 2-溴棕榈酸在制备防治骨丢失相关疾病的药物中的应用 (Application of 2-bromopalmitic acid in preparation of medicine for preventing and treating bone loss related diseases ) 是由 陈建权 章礼炜 杨惠林 于 2020-08-31 设计创作，主要内容包括：本发明公开了一种2-溴棕榈酸在制备防治骨丢失相关疾病的药物中的应用,属于医药技术领域。本发明利用抗酒石酸酸性磷酸酶染色、鬼笔环肽荧光染色、骨板骨吸收功能评价、逆转录实时荧光定量PCR、蛋白质免疫印迹等多种实验手段,首次明确2-BP可抑制BMMs在RANKL刺激下破骨细胞分化过程。并通过构建小鼠去卵巢骨质疏松模型,明确2-BP在体内也具有抑制破骨细胞分化、活化的功能,可显著缓解雌激素缺失导致的大量骨丢失病理过程,提供了其可防治骨丢失相关疾病的体内实验数据基础。(The invention discloses an application of 2-bromopalmitic acid in preparation of a medicine for preventing and treating bone loss related diseases, and belongs to the technical field of medicines. The invention utilizes a plurality of experimental means such as tartaric acid resistant acid phosphatase staining, phalloidin fluorescent staining, bone plate bone absorption function evaluation, reverse transcription real-time fluorescent quantitative PCR, protein immunoblotting and the like, and firstly confirms that 2-BP can inhibit the osteoclast differentiation process of BMMs under the stimulation of RANKL. And by constructing a mouse ovariectomized osteoporosis model, the 2-BP also has the functions of inhibiting osteoclast differentiation and activation in vivo, can obviously relieve a large number of bone loss pathological processes caused by estrogen deficiency, and provides an in vivo experimental data basis for preventing and treating bone loss related diseases.)

1. An application of 2-bromopalmitic acid in preparing medicine for preventing and treating bone loss related diseases is disclosed.

2. The use according to claim 1, wherein the bone loss related disease is osteoporosis, wear particle mediated osteolysis, rheumatoid arthritis or tumor bone destruction.

3. The use according to claim 1, wherein the agent for preventing or treating a disease associated with bone loss is an agent for inhibiting differentiation and maturation of osteoclasts.

4. The use according to claim 1, wherein the medicament for preventing or treating bone loss related diseases is a medicament for inhibiting the expression level of a characteristic gene during osteoclast differentiation.

5. The use of claim 4, wherein the signature genes comprise one or more of Nfatc1, C-fos, Ctsk, Acp5, Oscar, Dc-stamp, Atp6v0a3, Atp6v0d 2.

6. The use according to claim 1, wherein the medicament for preventing or treating bone loss related diseases is a medicament for inhibiting activation of c-Fos/NFATc1 signaling pathway during osteoclast differentiation.

7. The use of claim 1, wherein the medicament for preventing and treating bone loss related diseases is in the form of injection, capsule, oral preparation, or microcapsule.

8. The use according to claim 1, wherein the medicament for the prevention or treatment of a disease associated with bone loss is administered in an amount of 1 to 10 mg/kg.

Technical Field

The invention relates to application of 2-bromopalmitic acid in preparation of a medicine for preventing and treating bone loss related diseases, and belongs to the technical field of medicines.

Background

Bones are important components of the locomotor system, provide structural support and protection for the organism, and participate in physiological processes such as hematopoiesis, calcium metabolism, endocrine regulation and the like in vivo. Bone remodeling runs throughout the life of a human and most diseases related to bone metabolism affect the bone remodeling process. Bone remodeling in the normal body is dependent on the direct dynamic balance between Osteoclast (osteoplast) -mediated bone resorption and Osteoblast (osteoplast) -mediated bone formation, and imbalance between the two causes bone over-formation, sclerosis-related diseases or bone under-formation, over-resorption-related diseases. Osteoporosis (osteoporotis) is a systemic bone disease in which bone brittleness is increased due to decrease in bone density and bone quality caused by various reasons and destruction of bone microstructure, so that fracture easily occurs, and is closely related to aging of the body. In 2006, nearly 7000 million patients with osteoporosis in China have more than 2 hundred million people with osteopenia, and despite the lack of latest epidemiological data, the number of people who estimate osteoporosis and osteopenia in China has far exceeded the above numbers.

Osteoporosis has serious consequences, namely the occurrence of osteoporotic fracture, which is extremely harmful and is one of the main causes of disability and death of old patients. Epidemiological investigation based on the imaging shows that the prevalence rate of vertebral fracture of women over 50 years old in China is about 15%, the prevalence rate of vertebral fracture increases with increasing age after 50 years old, and the incidence rate of osteoporotic vertebral fracture of old women over 80 years old can reach 36.6%; while hip fracture is the most severe osteoporotic fracture, studies have shown that within 1 year after hip fracture occurs, 20% of patients die of various complications, about 50% of patients are disabled, the quality of life is significantly reduced, and a heavy burden is placed on the family and society. In addition, osteoclast and bone absorption mediated by osteoclast play an important role in the occurrence and development process of osteoporosis, are also important in diseases of bone mass loss such as periprosthetic osteolysis, rheumatoid osteoarthritis, tumor bone metastasis and the like, and are important targets for preventing and treating the diseases.

Currently, prevention of osteoporosis is mainly to regulate lifestyle, calcium agents, vitamin D, and its treatment is mainly focused on inhibiting osteoclast-mediated bone resorption or (and) promoting osteoblast-mediated bone formation. The bisphosphonates is the most widely clinically applied anti-osteoporosis drug at present, has high affinity with bone hydroxyapatite, can be specifically bound to the surface of active bone reconstruction, and inhibits osteoclast function so as to inhibit bone resorption. However, the use of bisphosphonates presents adverse gastrointestinal reactions, transient "flu-like" symptoms, renal toxicity and the risk of developing mandibular necrosis, atypical femoral fractures. Anti-osteoporosis drugs such as calcitonin, estrogen receptor modulator (raloxifene), menopausal hormone therapy, parathyroid hormone analogue (teriparatide) and RANKL monoclonal antibody (denosumab) have different indications, usages, dosages and side effects, and part of the drugs are high in price and difficult to be widely applied. Therefore, it is important to develop new drugs for inhibiting osteoclast differentiation and maturation.

Disclosure of Invention

In order to solve the problems, the invention provides application of 2-bromopalmitic acid in preparing a medicament for preventing and treating bone loss related diseases.

The invention aims to provide application of 2-bromopalmitic acid in preparation of a medicine for preventing and treating bone loss related diseases.

Further, the bone loss related disease is osteoporosis, wear particle mediated osteolysis, rheumatoid arthritis or tumor bone destruction.

Furthermore, the medicine for preventing and treating the diseases related to the bone loss is a medicine for inhibiting the differentiation and maturation of osteoclast.

Furthermore, the medicine for preventing and treating the bone loss related diseases is a medicine for inhibiting the expression level of characteristic genes in the differentiation process of osteoclasts.

Further, the characteristic gene comprises one or more of Nfatc1, C-fos, Ctsk, Acp5, Oscar, Dc-stamp, Atp6v0a3 and Atp6v0d 2.

Furthermore, the medicine for preventing and treating the diseases related to the bone loss is a medicine for inhibiting the activation of a c-Fos/NFATc1 signaling pathway in the differentiation process of osteoclasts.

Furthermore, the medicament for preventing and treating the bone loss related diseases is in the dosage form of injection, capsule, oral preparation and microcapsule preparation.

Furthermore, the administration dosage of the medicine for preventing and treating the bone loss related diseases is 1-10 mg/kg.

The invention has the beneficial effects that:

the invention utilizes a plurality of experimental means such as tartaric acid resistant acid phosphatase staining, phalloidin fluorescent staining, bone plate bone absorption function evaluation, reverse transcription real-time fluorescent quantitative PCR, protein immunoblotting and the like, and firstly confirms that 2-BP can inhibit the osteoclast differentiation process of BMMs under the stimulation of RANKL. And by constructing a mouse ovariectomized osteoporosis model, the 2-BP also has the functions of inhibiting osteoclast differentiation and activation in vivo, can obviously relieve a large number of bone loss pathological processes caused by estrogen deficiency, and provides an in vivo experimental data basis for preventing and treating bone loss related diseases.

Drawings

FIG. 1 is a CCK-8 method for detecting cytotoxicity of 2-BP to monocyte-macrophage;

FIG. 2 is an anti-tartrate acid phosphatase staining assay for the effect of 2-BP on the process of osteoclastic differentiation of monocytes macrophages;

FIG. 3 is a graph showing the effect of phalloidin fluorescence staining to detect the generation of F-activating protein characteristic of 2-BP on the osteoclastic differentiation process of monocyte macrophage;

FIG. 4 is a reverse transcription real-time fluorescence quantitative PCR detection of the expression of the related characteristic genes in the process of osteoclast differentiation;

FIG. 5 shows the effect of Western blotting on the expression of a transcription factor involved in the differentiation process of osteoclasts by 2-BP;

fig. 6 is a three-dimensional image of tibial cancellous bone and relevant bone mass parameters are analyzed.

Detailed Description

The present invention is further described below in conjunction with the following figures and specific examples so that those skilled in the art may better understand the present invention and practice it, but the examples are not intended to limit the present invention.