Application of malaviif in preparation of medicine for treating osteoporosis, malaviif sustained release system and application of malaviif sustained release system

Abstract

The invention provides application of malavirol in preparation of a medicine for treating osteoporosis, a malavirol sustained-release system and application of the malavirol sustained-release system, and relates to the technical field of medicines. The research of the inventor finds that malaviif restores the damaged macrophage intercellular function caused by OVX, removes apoptotic cells and reduces the release of inflammatory factors by antagonizing a CCR5 receptor and inhibiting the activation of a downstream signal channel of the CCR5 receptor, so that the inflammatory microenvironment in a marrow cavity is relieved, the bone homeostasis is recovered, the overexpression of osteoclasts is inhibited, and the osteoclast effect is improved. The composition is used for treating osteoporosis. According to the malaviif sustained release system provided by the invention, a drug library can be formed through subcutaneous injection, continuous release of malaviif is realized, the clinical compliance problems that the half-life period of malaviif is short and frequent administration is needed are solved, and a preferable medication scheme is provided for chronic disease management.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to the application of maraviro in the preparation of drugs for treating osteoporosis, a maraviro sustained-release system and its application. Background Technology

[0002] Osteoporosis is a systemic skeletal disease characterized by decreased bone mass, destruction of bone microstructure, leading to decreased bone strength, increased bone fragility, and susceptibility to fractures. In normal bone metabolism, osteoblast bone formation and osteoclast bone resorption are in dynamic equilibrium. When bone resorption exceeds bone formation, bone loss accelerates, bone density decreases, and osteoporosis ensues.

[0003] Currently, osteoporosis treatment primarily targets two aspects: inhibiting bone resorption and promoting bone formation. Teriparatide, a representative drug for promoting bone formation, is expensive and carries the risk of causing osteosarcoma. Estrogen-based drugs can inhibit bone resorption and reduce bone turnover in postmenopausal women, but long-term use has significant side effects. Bisphosphonates are contraindicated in patients with poor renal function, osteonecrosis of the mandible, and gastrointestinal irritation.

[0004] Therefore, it is necessary to explore a new drug that can be used for the treatment of osteoporosis.

[0005] In view of this, the present invention is hereby proposed. Summary of the Invention

[0006] The primary objective of this invention is to provide the use of Maraviroc (MVC) in the preparation of drugs for treating osteoporosis.

[0007] A second objective of this invention is to provide the use of Malawizo in the preparation of a medicament for repairing the impaired cytotoxicity of macrophages.

[0008] A third objective of this invention is to provide the use of malaviro in the preparation of a drug that inhibits osteoclast formation.

[0009] The fourth objective of this invention is to provide a Malawizo sustained-release system.

[0010] The fifth objective of this invention is to provide the application of the above-mentioned Malawiro sustained-release system in the preparation of drugs for treating osteoporosis.

[0011] To achieve the above objectives, the following technical solution is adopted: In a first aspect, the present invention provides the use of Malawiro in the preparation of drugs for treating osteoporosis.

[0012] As a further technical solution, the osteoporosis includes postmenopausal osteoporosis.

[0013] As a further technical solution, the drug also includes a carrier for loading maraviro.

[0014] As a further technical solution, the carrier includes a hydrogel.

[0015] As a further technical solution, the dosage form of the drug includes tablets, capsules, or injections.

[0016] Secondly, this invention provides the application of Malawiro in the preparation of drugs that repair the damaged cytotoxicity of macrophages.

[0017] Thirdly, the present invention provides the use of Malawiro in the preparation of drugs that inhibit osteoclast formation.

[0018] Fourthly, the present invention provides a maraviro sustained-release system comprising maraviro and a hydrogel; The hydrogel encapsulates Malawijo; The hydrogel is mainly composed of polycarboxymethyl methacrylate (PCM), poloxamer 407, and water.

[0019] As a further technical solution, in the hydrogel, the mass percentage of polycarboxymethyl methacrylate (PCM) is 0.025% and the mass percentage of poloxamer 407 is 26%.

[0020] Fifthly, the present invention provides the application of the above-mentioned Malawiro sustained-release system in the preparation of drugs for treating osteoporosis.

[0021] Compared with the prior art, the present invention has the following beneficial effects: The inventors discovered that Malawizo inhibits the activation of downstream signaling pathways by antagonizing the CCR5 receptor, thereby restoring the damaged macrophage burial function caused by OVX, clearing apoptotic cells, reducing the release of inflammatory factors, alleviating the inflammatory microenvironment in the bone marrow cavity, restoring bone homeostasis, and inhibiting excessive osteoclast production, thus making it suitable for the treatment of osteoporosis.

[0022] Malawiro is hydrophobic and metabolized rapidly, and can only be used orally in clinical practice. The Malawiro sustained-release system provided by this invention can form a "drug reservoir" through subcutaneous injection, achieving continuous release of Malawiro. This solves the clinical compliance problem of Malawiro's short half-life and the need for frequent dosing, providing an optimal medication regimen for chronic disease management. Attached Figure Description

[0023] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0024] Figure 1 CT scan of the distal femur in mice (Sham: sham surgery group, OVX: castration surgery model, OVX+MVC: castration surgery model + intraperitoneal injection of Malawizo). Figure 2 CCR5 expression levels in different subsets of monocytes and macrophages, grouped as follows: Figure 1 ; Figure 3 KEGG enrichment map of upregulated genes in regulatory macrophages of the OVX group and OVX+MVC group; Figure 4 MerTK expression levels in different subsets of monocytes and macrophages, grouped as follows: Figure 1 ; Figure 5 The ability of macrophages in the bone and femoral medullary cavity of mice in three groups—sham surgery group, castration surgery model group, and castration surgery model group + Malawizo intraperitoneal injection treatment group—to phagocytose apoptotic cells; Figure 6 The ratio of M1 and M2 macrophages in the femoral medullary cavity of mice in three groups: sham surgery group, castration surgery model, and castration surgery model + Malawizo injection treatment. Figure 7 MerTK expression during primary mononuclear macrophage-induced osteoclastogenesis (M-CSF: control group, M-CSF+RANKL: conditions for macrophage induction, M-CSF+RANKL+MVC: addition of maraviro to induce osteoclastogenesis). Figure 8 The ability of macrophages to engulf apoptotic cells during primary mononuclear macrophage-induced osteoclastogenesis; Figure 9 Primary mononuclear macrophage-induced osteoclastogenesis and its induction after addition of Malawi et al. Figure 10 The release of Malawiro in the Malawiro sustained-release system; Figure 11 CT scan of the distal femur of mice (Sham: sham surgery group, OVX: castration surgery model, Maraviroc@Gel: castration surgery model + subcutaneous injection of Maraviroc sustained-release system in the back, once a week). Detailed Implementation

[0025] The embodiments and examples of the present invention will be described in detail below. However, those skilled in the art will understand that the following embodiments and examples are for illustrative purposes only and should not be considered as limiting the scope of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention. Unless otherwise specified, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0026] In a first aspect, the present invention provides the use of Malawiro in the preparation of drugs for treating osteoporosis.

[0027] The inventors discovered that Malawizo inhibits the activation of downstream signaling pathways by antagonizing the CCR5 receptor, thereby restoring the damaged macrophage burial function caused by OVX, clearing apoptotic cells, reducing the release of inflammatory factors, alleviating the inflammatory microenvironment in the bone marrow cavity, restoring bone homeostasis, and inhibiting excessive osteoclast production, thus making it suitable for the treatment of osteoporosis.

[0028] In this invention, osteoporosis includes postmenopausal osteoporosis.

[0029] In some alternative embodiments, the drug further includes a carrier for loading maraviro.

[0030] The delivery or sustained release of Malawiro can be achieved through a carrier.

[0031] In some alternative embodiments, the carrier includes, but is not limited to, hydrogels, or other drug carriers known to those skilled in the art.

[0032] In some alternative embodiments, the dosage form of the drug includes, but is not limited to, tablets, capsules, or injections.

[0033] Secondly, this invention provides the application of Malawiro in the preparation of drugs that repair the damaged cytotoxicity of macrophages.

[0034] The inventors discovered that Malawi has the ability to repair macrophage damage caused by osteoclasts.

[0035] Thirdly, the present invention provides the use of Malawiro in the preparation of drugs that inhibit osteoclast formation.

[0036] The inventors discovered that Malawiro has the effect of inhibiting osteoclast formation.

[0037] Fourthly, the present invention provides a maraviro sustained-release system comprising maraviro and a hydrogel; The hydrogel encapsulates Malawijo; The hydrogel is mainly composed of polycarboxymethyl methacrylate (PCM), poloxamer 407, and water.

[0038] The maraviro sustained-release system provided by this invention can form a "drug reservoir" through subcutaneous injection to achieve continuous release of maraviro, solving the clinical compliance problem of short half-life and frequent dosing required by maraviro, and providing an optimal medication regimen for chronic disease management.

[0039] In some alternative embodiments, the hydrogel contains 0.025% polycarboxymethyl methacrylate and 26% poloxamer 407 by mass.

[0040] In some optional embodiments, the preparation method of the Malawizo sustained-release system is as follows: Preparation of thermosensitive hydrogel: Polycarbamide (PCB) was weighed and slowly added to water under stirring conditions, with continuous stirring until the compound was completely dispersed; then poloxamer 407 (P407) was added at 4°C under stirring conditions, and stirring was continued for 24 hours. A P407 / PCB (26% / 0.025%) hydrogel was obtained. After completion, the thermosensitive hydrogel was stored at 4°C.

[0041] Maraviroxine encapsulated in hydrogel: Maraviroxine was completely dissolved in DMSO and PEG300 (1:1). It was then injected into a hydrogel to prepare a maraviroxine sustained-release system with a final concentration of 10 mg / ml.

[0042] Fifthly, the present invention provides the application of the above-mentioned Malawiro sustained-release system in the preparation of drugs for treating osteoporosis.

[0043] The Malaviro sustained-release system provided by this invention includes Malaviro and has the effect of treating osteoporosis.

[0044] The present invention will be further illustrated below with specific embodiments. However, it should be understood that these embodiments are merely for the purpose of more detailed illustration and should not be construed as limiting the present invention in any way.

[0045] Example 1 A Malawio sustained-release system is prepared as follows: Preparation of thermosensitive hydrogel: Polycarbamide (PCB) was weighed and slowly added to water under stirring conditions, with continuous stirring until the compound was completely dispersed; then poloxamer 407 (P407) was added at 4°C under stirring conditions, and stirring was continued for 24 hours. A P407 / PCB (26% / 0.025%) hydrogel was obtained. After completion, the thermosensitive hydrogel was stored at 4°C.

[0046] Maraviroxine encapsulated in hydrogel: Maraviroxine was completely dissolved in DMSO and PEG300 (1:1). It was then injected into a hydrogel to prepare a maraviroxine sustained-release system with a final concentration of 10 mg / ml.

[0047] Experimental Example 1 A mouse model of postmenopausal osteoporosis (PMOP) was established using the castration (OVX) method. Two weeks after surgery, mice were administered maraviroxine (10 mg / kg / day) for four weeks, designated as the OVX+MVC group. A sham surgery group and an OVX model group were also included. MicroCT results showed that MVC effectively reduced OVX-induced bone loss. Figure 1 Similarly, six weeks post-surgery, bone marrow cells were extracted from mice, and CD45+ cells and CD45+CD11b+F4 / 80+ cells were sorted and sequenced 1:1. Sequencing data showed that CCR5 was mainly expressed in regulatory macrophages of OVX mice. Figure 2 KEGG pathway enrichment of significant genes in this group of cells (regulatory macrophages) indicated enhanced cell burial pathway. Figure 3 Furthermore, MerTK (a key receptor for the cell demise pathway) expression in this cell population was impaired in the OVX group, while MVC treatment promoted MerTK expression. Figure 4 ). Using the same mouse samples from each group (OVX+MVC, Sham, OVX), bone marrow macrophages were extracted and co-cultured with PI-labeled apoptotic Jurakat cells for 2 hours. Macrophages were then labeled with CD11b and F4 / 80 and analyzed. Flow cytometry showed that, compared with the control group, the OVX group mice had impaired macrophage burial capacity, while MVC could repair the impaired macrophage burial capacity in OVX mice. Figure 5 Further flow cytometry experiments confirmed that after MVC treatment, the proportion of M1 / M2 macrophages in the bone marrow cavity of OVX mice decreased. Figure 6 This indicates that the inflammatory environment in the bone marrow cavity was suppressed and the immune microenvironment was remodeled. Similarly, we extracted bone marrow macrophages from 6-week-old, normal C57 / bl mice and cultured them in vitro using 50 ng / ml RANKL and 30 ng / ml M-CSF to simulate osteoclastogenesis. On day 2 of culture, 100 μm of MVC was added, and samples were collected on day 3 for MerTK fluorescence staining. The results showed that, compared with the control group, under osteoclastogenic conditions, MerTK expression in macrophages was suppressed. The addition of MVC restored its expression (…). Figure 7On day 3, samples were collected for cell burial experiments. Apoptotic Jurkat cells (10w / 24 wells) were co-cultured with BMMs for 2h, followed by fixation and staining. The results showed that, compared with the control group, macrophage cell burial ability was impaired under osteoclastogenic conditions, while MVC could repair its impaired cell burial ability. Figure 8 Samples were collected on the fourth day of induction (using the same experimental method as above, but sampled on the fourth day) and subjected to TRAP staining to observe osteoclastogenesis. TRAP staining results showed that MVC could inhibit osteoclastogenesis. Figure 9 The above results also confirm that MVC enhances MerTK expression to repair damaged macrophage burial capacity and inhibit osteoclastogenesis. In vitro and in vivo experiments revealed a novel, non-chemotactic function of CCR5 in the bone immune microenvironment: restoring bone homeostasis and increasing bone mass by repairing damaged macrophage burial function.

[0048] We prepared an MVC sustained-release system using hydrogel technology. The drug-loaded hydrogel (MVC sustained-release system) was placed in water and stirred at a constant speed of 37°C. The solution concentration was measured daily until drug release ceased. The cumulative drug release was calculated based on the drug concentration at different time points, and a drug release curve was plotted. In vitro experiments confirmed that this system can continuously release the drug for more than 7 days. Figure 10 Using the same OVX-induced PMOP model, we injected the system subcutaneously into the backs of mice two weeks post-surgery, once a week for four consecutive weeks, before collecting samples. MicroCT results showed that, compared with the OVX group, this sustained-release system effectively reduced OVX-induced bone loss. Figure 11 This solves the clinical compliance challenges posed by the short half-life of MVC and the need for frequent dosing, providing an optimal medication regimen for chronic disease management.

[0049] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of Maraviroc in the preparation of a medicament for treating osteoporosis.

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

3. Use according to claim 1, characterized in that, The medicament further comprises a carrier for loading the maraviroc.

4. Use according to claim 3, characterized in that, The carrier comprises a hydrogel.

5. The use according to claim 1, characterized in that, The dosage form of the medicament comprises tablets, capsules or injections.

6. Use of Maraviroc in the preparation of a medicament for repairing the impaired phagocytosis of macrophages.

7. Use of Maraviroc in the preparation of a medicament for inhibiting the generation of osteoclasts.

8. A maraviroc sustained release system, characterized in that, The carrier comprises a hydrogel; The hydrogel encapsulates the maraviroc; The hydrogel mainly consists of polycarbophil, poloxamer 407 and water.

9. The maraviroc sustained release system according to claim 8, wherein, In the hydrogel, the mass percentage of polycarbophil is 0.025%, and the mass percentage of poloxamer 407 is 26%.

10. Use of the maraviroc sustained-release system of claim 8 or 9 in the preparation of a medicament for treating osteoporosis.

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