Application of securinine in preparation of long-term anti-osteoporosis medicine

Abstract

The invention discloses an application of securinine in preparation of an anti-osteoporosis medicine. A mouse model with osteoporosis is treated by securinine for a long time, and it is found that bone loss can be effectively reversed, and the bone microstructure can be remarkably improved. The invention provides a treatment scheme for non-inflammatory osteoporosis through animal verification, and fills the blank of the prior art. The securinine can significantly improve bone quality after treatment, has small toxic and side effects, and can play a long-term therapeutic effect.

Description

Technical Field

[0001] This invention relates to the field of pharmaceuticals, and in particular to the application of eugenol in the preparation of long-term anti-osteoporosis drugs. Background Technology

[0002] Osteoporosis is a systemic skeletal disease characterized by decreased bone mass, deterioration of bone microstructure, and increased bone fragility, with a typical consequence being an increased risk of fragility fractures. The development of this disease is regulated by both congenital and acquired factors: congenital factors include race, sex, age, and family history; acquired factors include medication use, underlying diseases, nutritional status, and lifestyle. Among these, advanced age, postmenopausal estrogen decline in women, and decreased androgen secretion due to male sexual dysfunction are the main risk factors for developing the disease.

[0003] Currently, first-line anti-osteoporosis drugs mainly include bisphosphonates, denosumab, teriparatide, and selective estrogen receptor modulators, but they have certain limitations and side effects. Denosumab, a first-line drug, is a monoclonal antibody targeting RANKL, which can effectively inhibit osteoclast-mediated bone resorption by blocking the RANKL-RANK interaction. However, its clinical application carries potential risks. Studies have shown that this drug can induce hypocalcemia and cause severe anxiety in patients, and there are also case reports of mood disorders such as anxiety and depression after drug use without accompanying calcium and phosphorus metabolism imbalance. This indicates that the mechanism by which denosumab affects the central nervous system is complex and may be independent of calcium homeostasis disturbances, bringing complexity and uncertainty to the treatment of osteoporosis patients with comorbid mood disorders.

[0004] Therefore, the purpose of this invention is to overcome the aforementioned deficiencies in the prior art and provide a novel treatment regimen for the safe and effective long-term treatment of osteoporosis. Based on this, we have discovered that the natural alkaloid securinine, under specific long-term dosing regimens, can effectively improve bone microstructure, providing a new option for the treatment of osteoporosis.

[0005] Securinine is a natural alkaloid extracted from plants of the genus *Securinine*. Recent studies have shown that securinine has potential in bone metabolism; however, current technologies mainly focus on the short-term anti-bone resorption effects of securinine in inflammatory models. No publicly available technology has yet revealed the therapeutic effects of securinine in long-term administration models of non-inflammatory (e.g., hormone-induced) osteoporosis, particularly its reversal effects on bone microstructure (e.g., trabecular bone thickness, bone volume fraction) and optimal dosage regimens over long-term treatment. Summary of the Invention

[0006] To address the shortcomings of existing technologies, this invention proposes the application of eugenol in the preparation of long-term anti-osteoporosis drugs.

[0007] This invention provides the application of eugenol in the preparation of an anti-osteoporosis drug. In some embodiments, treatment of bilaterally ovariectomized mice with eugenol can alleviate osteoporosis in the mice. Bilaterally ovariectomized (OVX) mice are a classic and commonly used animal model for studying osteoporosis. Ovariectomy leads to a decrease in estrogen levels in mice, inducing osteoporosis, and is particularly suitable for simulating the pathological process of osteoporosis in postmenopausal women.

[0008] Furthermore, the osteoporosis includes postmenopausal osteoporosis.

[0009] Furthermore, the dosage of the cyclophosphamide is 0.5-20 mg / kg / day.

[0010] Furthermore, the dosage of the sucralose is 5 mg / kg / day. In some embodiments, long-term oral administration of 5 mg / kg / day of sucralose to ovariectomized mice for 6 weeks showed a clear therapeutic effect.

[0011] Furthermore, the dosing cycle of the aforementioned cyclophosphamide is 6 weeks.

[0012] The present invention also provides a pharmaceutical composition for treating osteoporosis, the pharmaceutical composition comprising the aforementioned eugenol.

[0013] Furthermore, the pharmaceutical composition also includes a pharmaceutically acceptable carrier and / or excipients.

[0014] Furthermore, the excipient is any one of sucrose, starch, dextrin, talc, magnesium stearate, and calcium sulfate.

[0015] Furthermore, the dosage form of the pharmaceutical composition is any one of capsules, tablets, powders, and granules.

[0016] Furthermore, the formulation is an oral formulation, and compared to drugs that require injection, long-term use of oral formulations results in better patient compliance.

[0017] In summary, compared with the prior art, the present invention achieves the following technical effects: (1) The solution is clear: This invention provides a long-term dosage and course of treatment for non-inflammatory osteoporosis that has been verified by animal experiments, filling the gap in the prior art; (2) Significant therapeutic effect: This invention confirms that long-term administration of Securinine can not only inhibit bone resorption, but also significantly reverse the trabecular bone structure and improve bone quality, with clear effects; (3) High safety profile: This invention clarifies the effective and safe dosage range of securinine under long-term (up to six weeks) administration. Its natural product properties also suggest a lower risk of toxic side effects; (4) Good compliance: This invention has demonstrated the effectiveness of oral gavage administration, and patients have better compliance with long-term use compared to drugs that require injection. Attached Figure Description

[0018] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0019] Figure 1 Figure 1 shows the results of the construction and verification of the bilateral ovariectomy model in mice according to Example 1 of the present invention; (A) is a schematic diagram of bilateral ovariectomy in C57 mice; (B) is a figure showing the weight statistics of mice in the OVX group and sham group; (C) is a figure showing the bone mineral density statistics of mice in the OVX group and sham group before and after modeling.

[0020] Figure 2 The following figures illustrate the efficacy and dosage verification results of long-term Securinine treatment in osteoporotic mice as shown in Example 2 of this invention: (A) Bone structure diagrams under Micro-CT of the OVX group and sham-operated group treated with Securinine at 0 mg / kg and 5 mg / kg; (B) Micro-CT indicators of mice under different Securinine concentrations, including BV / TV and Tb.Th; (C) Statistical results of the level of type I collagen C-terminal peptide (CTX-1) in mouse serum after administration of different concentrations of Securinine. Detailed Implementation

[0021] To enable those skilled in the art to better understand the present invention, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.

[0022] Example 1: Construction of an osteoporosis model Experimental methods (1) Animal model construction Nine-week-old female C57BL / 6 mice were randomly divided into a sham-operated group and an ovariectomy group. The OVX group underwent bilateral ovariectomy to establish a mouse model of osteoporosis. Figure 1 As shown in A; in the Sham group, only the ovaries were exposed but not removed. All mice were fed in the same environment for 4 weeks to stabilize the model phenotype.

[0023] (2) Bone mineral density test Bone density was analyzed using the Medikors small animal body composition analyzer (InAlyzer) and dual-energy X-ray animal body composition analysis system. The samples were placed on the same horizontal plane and in the same direction for dual-energy X-ray scanning analysis to determine the overall decrease in bone density, thus proving the success of OVX modeling.

[0024] Experimental results Figure 1 B and Figure 1 C showed that, compared with the sham surgery group, mice after ovariectomy had a significant increase in body weight and a decrease in bone density compared with before modeling, indicating the successful establishment of the osteoporosis model.

[0025] Example 2: Efficacy and dosage verification of long-term treatment of osteoporosis with basil. Experimental methods (1) Administration by gavage Securinine was administered to OVX model mice at doses of 0.5, 1, 5, 10 mg / kg, and 20 mg / kg, with a drug-free solvent group (0 mg / kg) serving as a control. The mice were administered the drug orally via gavage for six weeks.

[0026] (2) Evaluation of the efficacy of securinine After the dosing cycle, the perfused animals or the separated leg bones were analyzed using VENUS Micro-CT (VENUS VNC-102S) to examine the bone microstructure of the distal femur. Key indicators included bone mineral density (BMD), bone volume fraction (BV / TV), and trabecular bone thickness (Tb.Th). BMD, BV / TV, and Tb.Th are important indicators for assessing bone health and osteoporosis, each reflecting skeletal condition from different perspectives.

[0027] (3) Detection of biochemical markers of bone transformation The ELISA kit detects serum C-terminal peptide (CTX-1) levels of type I collagen crosslinking. CTX-1 is an important bone metabolism marker reflecting bone resorption and is mainly used to assess osteoclast activity and the degree of bone resorption. It has important clinical value in the diagnosis, treatment monitoring, and fracture risk assessment of osteoporosis.

[0028] Experimental results As attached Figure 2 As shown in A and 2B, compared with the OVX control group, mice treated with 5 mg / kg Securinine for a long period of time showed significantly increased BV / TV and Tb.Th indices, effectively reversing OVX-induced bone loss. Figure 2 As shown in C, the detection of CTX-1 revealed that, compared with the untreated group, the serum CTX-1 level in the 5 mg / kg Securinine-treated group was significantly lower, indicating that it can effectively inhibit the bone resorption process.

[0029] The above experimental results indicate that long-term (6 weeks) oral administration of Securinine at a dose of 5 mg / kg / d can safely and effectively treat OVX-induced osteoporosis, and its mechanism is related to the inhibition of bone resorption.

[0030] This invention was validated through standardized in vivo pharmacodynamic experiments: in an OVX-induced mouse osteoporosis model, long-term oral administration of Securinine (5 mg / kg / day) for six weeks showed a clear therapeutic effect. Micro-CT bone morphometric analysis confirmed that this regimen significantly reversed bone loss and improved key indicators such as BMD and BV / TV; simultaneously, the significant reduction in serum bone resorption marker CTX-1 levels biochemically confirmed its effective inhibition of bone resorption, fully demonstrating the feasibility and significant efficacy of this technical solution.

[0031] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. Application of eugenol in the preparation of anti-osteoporosis drugs.

2. The application according to claim 1, characterized in that, The osteoporosis mentioned includes postmenopausal osteoporosis.

3. The application according to claim 1, characterized in that, The dosage of the cyclophosphamide is 0.5-20 mg / kg / day.

4. The application according to claim 1, characterized in that, The dosage of the cyclophosphamide is 5 mg / kg / day.

5. The application according to claim 4, characterized in that, The administration period for the cyclophosphamide is 6 weeks.

6. A pharmaceutical composition for treating osteoporosis, characterized in that, The pharmaceutical composition comprises the eugenol as described in claim 1.

7. The pharmaceutical composition according to claim 6, characterized in that, The pharmaceutical composition also includes a pharmaceutically acceptable carrier and / or excipients.

8. The pharmaceutical composition according to claim 7, characterized in that, The excipients are any one of sucrose, starch, dextrin, talc, magnesium stearate, and calcium sulfate.

9. The pharmaceutical composition according to claim 6, characterized in that, The pharmaceutical composition is formulated in any one of the following dosage forms: capsules, tablets, powders, and granules.

10. The pharmaceutical composition according to claim 9, characterized in that, The preparation is an oral preparation.

Images