Application of TXNIP specific inhibitor in preparation of product for preventing and treating osteoporosis or fracture

Abstract

The invention relates to application of a TXNIP specific inhibitor in preparation of a product for preventing and treating osteoporosis or fracture. The product is a composition or preparation containing SRI-37330 or an optical isomer thereof, or pharmaceutically acceptable salt thereof. Compared with the prior art, it is found that SRI-37330 can efficiently and specifically reduce the level of TXNIP protein in osteogenesis and osteoclast and inhibit differentiation of osteogenesis and osteoclast, and therefore the SRI-37330 has the functions of promoting bone metabolism to enter low conversion so as to prevent and treat diabetic osteoporosis, fracture and postmenopausal osteoporosis. The invention proves that the SRI-37330 has an active ingredient capable of safely and effectively promoting bone metabolism to enter low conversion so as to prevent and treat diabetic osteoporosis, fracture and postmenopausal osteoporosis.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular to the application of a TXNIP specific inhibitor in the preparation of a product for preventing and treating osteoporosis or bone fracture. Background Art

[0002] Osteoporosis is a systemic disease characterized by decreased bone mass and destruction of the microstructure of bone tissue, which leads to increased bone brittleness and susceptibility to fractures. Current research generally believes that it is a common age-related disease caused by the combined effects of genetic and environmental factors, and that peak bone mass and osteoporosis have a genetic tendency. Studies on twins have found that 50%-90% of the variation associated with bone density can be inherited; if parents have a history of osteoporotic fractures, the genetic risk of their children suffering from osteoporotic fractures is 1.30-4.24. Especially after menopause, the level of estrogen in women decreases significantly, leading to accelerated bone loss, which eventually causes postmenopausal osteoporosis, a common disease in postmenopausal women. With the aggravation of the aging of society, the osteoporosis of diabetic patients is becoming more and more serious, placing a huge burden on society and the economy.

[0003] With the development of large-scale single nucleotide polymorphism genotyping technology, the genome-wide association study of bone density has been greatly promoted. At present, a series of common genetic variants of osteoporosis have been successfully identified, but a large part of osteoporosis-related genetic variants still need to be further studied and discovered. In 2017, Nature Genetics analyzed more than 140,000 participants in the UK Biobank through a genome-wide association study. Combined with the patient's heel ultrasound bone density, 153 gene loci related to bone density were found. These gene loci account for 12% of the causes of genetic diseases. In 2018, Nature Genetics' research identified 518 genomic loci related to bone density (301 of which were newly discovered) through a study of more than 426,000 people in the UK Biobank, which can explain 20% of the genetic mutations associated with osteoporosis, and is expected to help researchers develop new targeted therapies to treat osteoporosis and reduce the risk of fractures in the population. However, genetic variants related to osteoporosis still need further research.

[0004] In previous basic research, it was found that the changes in bone metabolism status were particularly obvious during the pathophysiological process of osteoporosis. Therefore, genes that affect the metabolism, differentiation, maturation, and function of osteoblasts and osteoclasts are the key points in the research on the prevention and treatment of osteoporosis. Patent CN202011366096.5 discloses a bone formation-promoting polypeptide and its application. The polypeptide can enhance the differentiation, maturation, or calcification of osteoblasts or cells that can differentiate into osteoblasts, and is used for the treatment or prevention of bone diseases. The bone formation-promoting polypeptide has stable chemical properties, is easy to synthesize on a large scale at low cost, has high biocompatibility, and is non-toxic, and can be widely used for the prevention and treatment of bone diseases. Although the synthesis method of the polypeptide and some in vitro experiments are provided, there is a lack of large-scale clinical trial data to prove its safety and effectiveness in humans; and whether there are potential side effects or safety problems in long-term application in animal experiments is not mentioned.

[0005] Therefore, researching drugs that can be clinically applied to target related genes to promote bone metabolism transformation has very important clinical diagnosis and treatment and basic scientific research value for the prevention and treatment of osteoporosis. Summary of the Invention

[0006] The purpose of the present invention is to provide an application of a TXNIP specific inhibitor in the preparation of products for preventing and treating osteoporosis or fractures, which can be used to promote bone metabolism into low turnover to prevent and treat osteoporosis or fractures, especially diabetic osteoporosis, fractures, and postmenopausal osteoporosis.

[0007] The purpose of the present invention can be achieved by the following technical solutions:

[0008] In the first aspect of the present invention, an application of a TXNIP specific inhibitor in the preparation of products for preventing and treating osteoporosis or fractures is provided. The TXNIP specific inhibitor with oral activity, or its optical isomer, or its pharmaceutically acceptable salt is used to prepare a composition or preparation, and the composition or preparation is used to promote bone metabolism into low turnover to prevent and treat osteoporosis or fractures, especially diabetic osteoporosis, fractures, and postmenopausal osteoporosis, etc. The TXNIP specific inhibitor with oral activity is SRI-37330, and its chemical structural formula is as follows:

[0009]

[0010] In another preferred example, the composition or preparation is used for one or more of the following applications:

[0011] (i) Inhibiting the differentiation and absorption of osteoclasts and the differentiation and mineralization of osteoblasts, thereby promoting bone metabolism into a low turnover state;

[0012] (ii) Promoting bone metabolism into low turnover to delay the occurrence of osteoporosis and promote the healing of diabetic fractures;

[0013] (iii) Reduce the expression level of TXNIP in osteoblasts;

[0014] (iv) Reduce the expression level of TXNIP in osteoclasts. The above osteoblasts are somatic cells, selected from: bone marrow mesenchymal stem cells, osteoprogenitor cells, pre-osteoblasts, osteoblasts; osteoclasts are somatic cells, selected from bone marrow-derived mononuclear macrophages, pre-osteoclasts, osteoclasts.

[0015] In another preferred embodiment, the composition is a pharmaceutical composition, a health product composition, a dietary supplement or a food composition.

[0016] In another preferred embodiment, the preparation includes a dietary supplement, a food additive, or a test reagent.

[0017] In another preferred embodiment, the pharmaceutical composition contains (a) an orally active TXNIP-specific inhibitor or a pharmaceutically acceptable salt thereof as an active ingredient; and (b) a pharmaceutically acceptable carrier or excipient.

[0018] In another preferred embodiment, the pharmaceutical composition further includes additional reduction of glucagon secretion and action.

[0019] In another preferred embodiment, in the pharmaceutical composition, the content of the orally active TXNIP-specific inhibitor is 0.001 - 99 wt%, preferably 0.01 - 90 wt%, based on the total weight of the composition. Further preferably, the amount of effective SRI-37330 contained in the composition or preparation is 10 - 60 wt%.

[0020] In another preferred embodiment, the pharmaceutical composition is an injectable drug or an oral drug.

[0021] In another preferred embodiment, the form of the pharmaceutical composition is selected from the group consisting of: tablets, capsules, granules, suspensions, pills, solutions, syrups, or injections.

[0022] In another preferred embodiment, the composition or preparation is administered to a mammal.

[0023] In another preferred embodiment, the mammal includes: primates; more preferably the mammal is a human.

[0024] In another preferred embodiment, the health product composition contains an orally active TXNIP-specific inhibitor or a pharmaceutically acceptable salt thereof as an active ingredient, and promotes low-turnover bone metabolism to prevent diabetic osteoporosis and postmenopausal osteoporosis.

[0025] In another preferred embodiment, in the health care product composition, the content of the orally active TXNIP specific inhibitor is 0.001 - 99 wt%, preferably 0.01 - 90 wt%.

[0026] In a second aspect of the present invention, there is provided an application of a TXNIP specific inhibitor in the preparation of a product for preventing and treating osteoporosis or fractures. The application method includes the steps of:

[0027] Adding an orally active TXNIP specific inhibitor or its salt to a cell culture system, thereby reducing the expression level of TXNIP protein in the cells;

[0028] Or culturing cells in the presence of an orally active TXNIP specific inhibitor or its salt, thereby reducing the expression level of TXNIP protein in the cells.

[0029] In another preferred embodiment, in the above cell culture system, the concentration of the orally active TXNIP specific inhibitor is 0.01 - 100 uM, preferably 0.05 - 75 uM, more preferably 0.1 - 50 uM, and most preferably 0.5 - 10 uM.

[0030] In another preferred embodiment, the cells are eukaryotic cells.

[0031] In another preferred embodiment, the cells are derived from mammals, preferably human cells.

[0032] In another preferred embodiment, the cells are somatic cells.

[0033] In another preferred embodiment, the cells are selected from the group consisting of: bone marrow mesenchymal stem cells, osteoprogenitor cells, pre - osteoblasts, osteoblasts, bone marrow - derived macrophages, pre - osteoclasts, osteoclasts.

[0034] In a third aspect of the present invention, there is provided an application of a TXNIP specific inhibitor in the preparation of a product for preventing and treating osteoporosis or fractures, specifically including the steps of: administering an orally active TXNIP specific inhibitor, or its optical isomer, or its pharmaceutically acceptable salt to a subject in need as an active ingredient to promote bone metabolism into a low - turnover state for preventing and treating diabetic osteoporosis, fractures and post - menopausal osteoporosis.

[0035] In another preferred embodiment, the subject in need is a mammal, and the mammals include: primates; more preferably the mammal is a human.

[0036] In the fourth aspect of the present invention, there is provided a pharmaceutical composition for use of a TXNIP-specific inhibitor in the preparation of a product for preventing and treating osteoporosis or fractures, said pharmaceutical composition comprising: SRI-37330 (which can reduce the level of TXNIP).

[0037] (a1) An orally active TXNIP-specific inhibitor or a pharmaceutically acceptable salt thereof as an active ingredient, the active ingredient being SRI-37330;

[0038] (a2) An additional active ingredient that reduces the secretion and action of glucagon and blocks hepatic glucose output, the additional active ingredient being selected from the group consisting of: SRI-37330; tablets containing SRI-37330; oral agents containing SRI-37330.

[0039] (b) A pharmaceutically acceptable carrier or excipient.

[0040] In another preferred embodiment, in the said pharmaceutical composition, the content of the orally active TXNIP-specific inhibitor is 0.001-99 wt%, preferably 0.01-90 wt%, based on the total weight of the composition.

[0041] Compared with the prior art, the present invention has the following beneficial effects:

[0042] 1. The present invention first proves that the orally active TXNIP-specific inhibitor SRI-37330 can effectively inhibit osteoblast differentiation and mineralization, and inhibit osteoclast differentiation and resorption, ultimately promoting bone metabolism into low turnover to prevent and treat osteoporosis or fractures, especially fractures and postmenopausal osteoporosis. Existing SRI-37330 is generally used as an orally bioavailable thioredoxin-interacting protein (TXNIP) inhibitor, which inhibits glucagon secretion and function, reduces hepatic glucose production, and reverses hepatic steatosis. Although the impact on bone health of osteoporosis as a complication of diabetes has been discussed in the prior art, the treatment of diabetic osteoporosis is a complex process that requires comprehensive consideration of multiple factors. Although some existing diabetes drugs may have a positive impact on bone health, directly applying these methods to the treatment of osteoporosis still faces some technical difficulties and challenges. First, the pathophysiological mechanism is complex: there is a complex interaction between diabetes and osteoporosis. Diabetes can lead to osteoporosis not only by affecting multiple links of bone metabolism, but may also increase the risk of fractures by affecting bone quality. For example, even if the bone density of diabetic patients seems normal, they may be prone to fractures due to decreased bone quality. This indicates that the treatment of osteoporosis needs to be adjusted according to the specific pathophysiological mechanism of diabetes. Second, the impact of drugs: some hypoglycemic drugs may affect bone metabolism. For example, metformin can reduce the fracture risk in type 2 diabetic patients, but its specific mechanism has not been fully clarified. In addition, new hypoglycemic drugs such as GLP-1 receptor agonists have shown the potential to improve bone density and bone quality in animal experiments, but their long-term effects and safety in humans still need further study. Difficulty in evaluating treatment effects: the assessment of fracture risk usually relies on bone density measurement, but this method may underestimate the actual fracture risk in diabetic patients. Therefore, more accurate assessment tools are needed to predict the fracture risk in diabetic patients and guide treatment decisions. Finally, the individualization of treatment strategies: there is great heterogeneity in the bone health status of diabetic patients, and treatment strategies need to be individualized according to the specific situation of the patients. For example, multiple factors such as the type of diabetes, disease duration, complications, and drug use of the patients need to be considered. In summary, although existing diabetes treatment methods provide some inspiration for the treatment of osteoporosis, a series of technical and scientific challenges still need to be overcome to successfully apply these methods clinically. Future research needs to further explore the interaction between diabetes and osteoporosis, develop new treatment methods, and optimize existing treatment regimens.

[0043] 2. By inhibiting the specific gene TXNIP related to glucose metabolism and oxidative stress, orally active TXNIP-specific inhibitors exhibit the potential for preventing and treating diabetic osteoporosis, fractures, and postmenopausal osteoporosis, opening up a new research direction for more drugs to target osteoporosis by affecting specific genes related to metabolism and oxidative stress.

[0044] 3. The present invention discovers for the first time that an orally active TXNIP-specific inhibitor (or a composition and product of an orally active TXNIP-specific inhibitor) can be applied to promoting bone metabolism into low turnover for the prevention and treatment of diabetic osteoporosis, fractures, and postmenopausal osteoporosis products, which indicates the potential application value of SRI-37330 (or a composition and product of an orally active TXNIP-specific inhibitor) in osteoporosis.

[0045] 4. The orally active TXNIP-specific inhibitor selected in the present invention, or its optical isomer, or its pharmaceutically acceptable salt can be made into various pharmaceutical compositions, health product compositions, dietary supplements, food compositions, dietary supplements, food additives, test reagents, or cosmetic compositions, and administered to the required subjects in various ways, which is convenient to use. When the orally active TXNIP-specific inhibitor, or its optical isomer, or its pharmaceutically acceptable salt is made into various compositions, the selected carrier or excipient has no negative impact on SRI-37330.

[0046] 5. According to the severity of osteoporosis, the content of the orally active TXNIP-specific inhibitor in the composition or preparation is controlled to be 0.001 - 99 wt%. For the prevention of osteoporosis and promoting bone metabolism into low turnover, it can be gradually and adaptively added starting from 0.001%. For osteoporosis with clinical symptoms, when promoting bone metabolism into low turnover and restoring the lost bone mass, it may be necessary to add 99% in some cases. In special cases, consideration may be given to combining with other drugs.

[0047] 6. A TXNIP-specific inhibitor provided by the present invention has currently been widely applied in pre-clinical research, involving drug targets related to chronic diseases such as diabetes, and has long-term safety and effectiveness. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 It is a flow chart of OVX-induced osteoporosis and SRI37330 prevention experiment.

[0049] Figure 2These are the trabecular bone images of mouse tibia examined by micro-CT, including the blank control group (sham operation group), the surgical control group (OVX operation + PBS (phosphate buffer)), the low-concentration group (OVX + Low SRI-37330), and the high-concentration group (OVX + High SRI-37330).

[0050] Figure 3 This is the statistical chart of trabecular bone in mouse tibia examined by micro-CT.

[0051] Figure 4 These are the trabecular bone images of mouse femur examined by Von kossa.

[0052] Figure 5 This is the statistical chart of trabecular bone in mouse femur examined by Von kossa.

[0053] Figure 6 These are the trabecular bone images of mouse femur examined by double fluorescent labeling.

[0054] Figure 7 This is the statistical chart of bone formation rate indicated by double fluorescent labeling.

[0055] Figure 8 These are the TRAP staining images of mouse bone tissue sections.

[0056] Figure 9 This is the statistical chart of the TRAP staining results.

[0057] Figure 10 These are the experimental result images of treating diabetic fractures in vivo. Detailed implementation manners

[0058] The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments.

[0059] Through relevant in-depth research, the inventors unexpectedly discovered that an orally active TXNIP-specific inhibitor can inhibit osteoblast differentiation and mineralization, inhibit osteoclast differentiation and resorption, and ultimately promote bone metabolism to enter a low-turnover state to prevent and treat osteoporosis and fractures, especially fractures and postmenopausal osteoporosis, etc. Therefore, the orally active TXNIP-specific inhibitor has a safe and effective active ingredient that can promote bone metabolism to enter a low-turnover state to prevent and treat osteoporosis and fractures. Based on this, the present invention was completed.

[0060] Specifically, in the present invention, osteoblasts and osteoclasts were used as in vitro research objects, and mice were used as in vivo model animals. Through cytological experiments and experiments on constructing classic diabetic models and ovariectomized models of mice, it was confirmed that the orally active TXNIP specific inhibitor described in the present invention can inhibit osteoblast differentiation and mineralization; inhibit osteoclast differentiation and resorption; and ultimately promote bone metabolism to enter low turnover to prevent and treat diabetic osteoporosis, fractures, and postmenopausal osteoporosis.

[0061] The term

[0062] As used herein, the terms "active ingredient", "active ingredient for promoting bone metabolism to enter low turnover to prevent and treat diabetic osteoporosis, fractures, and postmenopausal osteoporosis", "composition", "composition or preparation for promoting bone metabolism to enter low turnover to prevent and treat diabetic osteoporosis, fractures, and postmenopausal osteoporosis" refer to an orally active TXNIP specific inhibitor, or a pharmaceutically acceptable salt thereof, or a solvate thereof, or an optical isomer thereof, or a racemate thereof, or a crystal thereof, or a hydrate thereof, or a deuterated compound thereof, or a combination thereof.

[0063] As used herein, the term "SRI-37330" refers to an orally active TXNIP specific inhibitor. The term "TXNIP" refers to Thioredoxin Interacting Protein (TXNIP). TXNIP is an important physiological inhibitor of the intracellular thioredoxin (TXN) redox system, is expressed in various organs and tissues, and plays a role in regulating intracellular redox reactions. In addition to playing an important role in redox regulation, TXNIP is also involved in various important physiological processes such as inflammatory reactions and glucose metabolism. Existing SRI-37330 is generally used as an orally bioavailable inhibitor of thioredoxin interacting protein (TXNIP), inhibits glucagon secretion and function, reduces hepatic glucose production, and reverses hepatic steatosis. However, there has been no technical report on SRI-37330 exerting the effect of promoting bone metabolism to enter low turnover to prevent and treat diabetic osteoporosis, fractures, and postmenopausal osteoporosis by inhibiting TXNIP.

[0064] Although SRI-37330 can inhibit glucagon secretion and thus reduce the body's blood glucose level, it is in two different fields from diabetic osteoporosis. Being able to treat diabetes does not necessarily mean being able to prevent and treat diabetic osteoporosis because, although the two diseases are interrelated, they are different. Diabetes can not only cause osteoporosis by affecting multiple links of bone metabolism but also increase the risk of fractures by affecting bone quality. Although some hypoglycemic drugs may have an impact on bone metabolism, the specific mechanism is still unclear and their long-term effectiveness remains to be investigated. Secondly, the impact of diabetic complications, such as nephropathy, retinopathy, and cardiovascular diseases, may also affect bone mass and bone quality during the treatment of these complications. Therefore, various complications need to be comprehensively considered to formulate a comprehensive treatment plan to ensure the mutual balance between the two. In addition, in addition to finding that SRI-37330 can promote bone metabolism into a low turnover state to delay the occurrence of osteoporosis and promote fracture healing, the present invention also finds that SRI-37330 inhibits the differentiation and absorption of osteoclasts and the differentiation and mineralization of osteoblasts, thus promoting bone metabolism into a low turnover state; generally, the treatment of osteoporosis mainly focuses on the conversion balance between osteoclasts and osteoblasts, including anti-resorption (inhibiting the activity of osteoclasts, such as bisphosphonates), bone formation promotion (enhancing the activity of osteoblasts, such as teriparatide), calcium and vitamin D supplementation, etc. The present invention not only promotes the differentiation and mineralization of osteoblasts but also inhibits the differentiation and absorption of osteoclasts, jointly promoting the increase of bone mass in two directions and being able to intervene in blood glucose changes, and is applicable to senile osteoporosis and postmenopausal osteoporosis.

[0065] Furthermore, the present invention can also reduce the expression level of TXNIP in osteoblasts and reduce the expression level of TXNIP in osteoclasts; improve the intracellular inflammatory and oxidative stress levels and stabilize the cell state.

[0066] As used herein, the term "bone metabolism" refers to the continuous cell metabolism of bone cells. Not only do the bone cells interact with each other, but there are also interactions between erythroid progenitor cells and stromal cells in the bone marrow to carry out bone remodeling and reconstruction.

[0067] As used herein, "promoting bone metabolism into a low turnover state to prevent and treat osteoporosis" means delaying, blocking, reducing, stopping, and / or reversing the process of bone mass loss.

[0068] Mouse osteoblasts (BMSC)

[0069] BMSC cells are differentiated from mesenchymal progenitor cells in the inner and outer periosteum and bone marrow stroma of mice and can specifically secrete a variety of bioactive substances to regulate and affect the process of bone formation and reconstruction.

[0070] In the pathophysiological process of osteoporosis, the changes in bone metabolism status are particularly obvious. Therefore, genes that affect the metabolism, differentiation, maturation, and function of osteoblasts and osteoclasts are the key points in the research on the prevention and treatment of osteoporosis.

[0071] Mouse osteoclasts (BMMs)

[0072] BMMs are the main functional cells of mouse bone resorption and play an important role in bone development, growth, repair, and reconstruction. Osteoclasts originate from the hematopoietic monocyte-macrophage system and are a special type of terminally differentiated cells. They can be formed by the fusion of their mononuclear precursors in various ways to form large multinucleated cells.

[0073] Mouse bilateral ovariectomy model (OVX model)

[0074] Surgical construction of the mouse bilateral ovariectomy (ovariectomy, OVX) model has been widely used in the mechanism research of osteoporosis.

[0075] Mouse diabetes model (DM model)

[0076] Constructing a diabetic mouse model by feeding a high-sugar and high-fat diet has been widely used in the mechanism research of diabetic osteoporosis.

[0077] Active ingredient

[0078] The active ingredient of the present invention for promoting bone metabolism to enter low turnover to prevent and treat diabetic osteoporosis, fractures, and postmenopausal osteoporosis is an orally active TXNIP-specific inhibitor or a pharmaceutically acceptable salt thereof.

[0079] SRI-37330 is an orally active TXNIP-specific inhibitor that can reduce the secretion and action of glucagon and block hepatic glucose output. Chemical formula: C 16 H 19 F3N4O2S, molecular weight: 388.41. The structural formula is as shown in the formula:

[0080]

[0081] In the present invention, a preferred active ingredient is a pharmaceutically acceptable compound of an orally active TXNIP-specific inhibitor.

[0082] As an orally active TXNIP-specific inhibitor, SRI-37330 has been widely studied for its ability to reduce the secretion and action of glucagon and block hepatic glucose output. SRI-37330 targets TXNIP and prevents and treats diabetic osteoporosis, fractures, and postmenopausal osteoporosis by simultaneously inhibiting the differentiation and resorption of osteoclasts and the differentiation and mineralization of osteoblasts to reduce bone metabolism.

[0083] Drug Compositions and Administration Methods

[0084] The present invention also provides a composition, preparation or product containing the active ingredient of the present invention, and the composition, preparation or product can be used to promote bone metabolism into low turnover to prevent and treat diabetic osteoporosis, fractures and postmenopausal osteoporosis. Representative compositions, preparations or products include drugs and health products that promote bone metabolism into low turnover to prevent and treat diabetic osteoporosis, fractures and postmenopausal osteoporosis.

[0085] A preferred composition is a pharmaceutical composition that contains an effective amount of an orally active TXNIP specific inhibitor or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0086] As used herein, the term "effective amount" or "effective dose" refers to an amount that can produce a function or activity (i.e., prevent and treat diabetic osteoporosis, fractures and postmenopausal osteoporosis) in humans and / or animals and is acceptable to humans and / or animals.

[0087] As used herein, the components of the term "pharmaceutically acceptable" are substances that are applicable to humans and / or mammals without excessive adverse side effects (such as toxicity, irritation and allergic reactions), that is, substances with a reasonable benefit / risk ratio. The term "pharmaceutically acceptable carrier" refers to a carrier used for administering a therapeutic agent, including various excipients and diluents.

[0088] The pharmaceutical composition of the present invention contains a safe and effective amount of the active ingredient of the present invention and a pharmaceutically acceptable carrier. Such carriers include (but are not limited to): saline, buffer solution, glucose, water, glycerol, ethanol, gel and their combinations. Usually, the pharmaceutical preparation should match the administration method, and the dosage form of the pharmaceutical composition of the present invention is injection, oral preparation (tablet, capsule, oral liquid), transdermal agent, sustained release agent. For example, it is prepared by a conventional method with physiological saline or an aqueous solution containing glucose and other adjuvants. The said pharmaceutical composition should be manufactured under aseptic conditions.

[0089] The effective amount of the active ingredient described in the present invention may vary depending on the mode of administration, the severity of the disease to be treated, and the like. The selection of the preferred effective amount can be determined by those of ordinary skill in the art based on various factors (e.g., through clinical trials). Such factors include, but are not limited to: the pharmacokinetic parameters of the active ingredient such as bioavailability, metabolism, half-life, etc.; the severity of the disease to be treated in the patient, the patient's body weight, the patient's immune status, the route of administration, etc. Generally, when the active ingredient of the present invention is administered at a dose of about 0.01 mg - 50 mg / kg of animal body weight per day (preferably 0.1 mg - 10 mg / kg of animal body weight), satisfactory effects can be obtained. For example, due to the urgency of the treatment situation, several separate doses may be administered per day, or the dose may be proportionally reduced.

[0090] Typically, when an orally active TXNIP-specific inhibitor is administered by local intra-articular injection, for a subject (human) with a body weight of 60 kg, the average daily dose is usually 0.6 - 3000 mg, preferably 6 - 600 mg, more preferably 45 - 90 mg.

[0091] The pharmaceutically acceptable carriers described in the present invention include (but are not limited to): water, saline, liposomes, lipids, peptides, cellulose, nanogels, or combinations thereof. The choice of carrier should match the mode of administration, which is well known to those of ordinary skill in the art.

[0092] A method for promoting bone metabolism into low turnover to prevent and treat diabetic osteoporosis, fractures, and postmenopausal osteoporosis:

[0093] The present invention provides a method for promoting bone metabolism into low turnover to prevent and treat diabetic osteoporosis, fractures, and postmenopausal osteoporosis (including therapeutic methods or in vitro non-therapeutic methods), as well as a method for inhibiting the expression and / or activity of TXNIP protein.

[0094] Typically, the method includes: adding the pharmaceutical composition or the active ingredient of the present invention to a cultured cell system, thereby delaying and / or reversing the pathological process of bone mass loss; or reducing the expression level of TXNIP protein.

[0095] In another preferred example, the cells are BMSC, BMMs cells, especially normal somatic cells.

[0096] The present invention also provides a method for promoting bone metabolism into low turnover to prevent and treat diabetic osteoporosis, fractures, and postmenopausal osteoporosis, including the step of administering to a subject in need an orally active TXNIP-specific inhibitor, or its optical isomer, or its pharmaceutically acceptable salt as an active ingredient for promoting bone metabolism into low turnover to prevent and treat diabetic osteoporosis, fractures, and postmenopausal osteoporosis.

[0097] Preferably, the method for promoting bone metabolism to enter low turnover to prevent and treat diabetic osteoporosis, fractures, and postmenopausal osteoporosis is to administer to the subject a health product containing an orally active specific inhibitor of TXNIP or a salt thereof.

[0098] The main advantages of the present invention include:

[0099] (a) The present invention for the first time demonstrates that an orally active specific inhibitor of TXNIP can effectively inhibit osteoblast differentiation and mineralization; inhibit osteoclast differentiation and resorption; and ultimately promote bone metabolism to enter low turnover to prevent and treat diabetic osteoporosis, fractures, and postmenopausal osteoporosis.

[0100] (b) The orally active specific inhibitor of TXNIP exhibits potential for the treatment of diabetic fractures and osteoporosis, opening up a new research direction for more drugs to target diabetic fractures and osteoporosis by affecting specific genes related to oxidative stress and metabolism.

[0101] (c) Through a convenient administration method, the orally active specific inhibitor of TXNIP exhibits a potential prospect for promoting and applying the repair of osteoporosis.

[0102] (d) TXNIP is an important physiological inhibitor of the intracellular thioredoxin (TXN) redox system, which is expressed in multiple organs and tissues and plays a role in regulating intracellular redox reactions. In addition to playing an important role in redox regulation, TXNIP is also involved in various important physiological processes such as inflammatory responses and glucose metabolism. SRI-37330 is an orally active specific inhibitor of TXNIP. SRI-37330 reduces the secretion and action of glucagon and blocks hepatic glucose output. SRI-37330 can be used in the research of obesity and diabetes. The present invention for the first time discovers that the orally active specific inhibitor RI-37330 of TXNIP (or a composition and product containing an orally active specific inhibitor of TXNIP) can be applied to promote bone metabolism to enter low turnover to prevent and treat diabetic osteoporosis, fractures, and postmenopausal osteoporosis, which indicates the potential application value of SRI-37330 (or a composition and product containing an orally active specific inhibitor of TXNIP) in the prevention and treatment of diabetic osteoporosis, fractures, and postmenopausal osteoporosis.

[0103] Therefore, the orally active specific inhibitor SRI-37330 of TXNIP of the present invention (or a composition and product containing an orally active specific inhibitor of TXNIP) can be applied to promote bone metabolism to enter low turnover to prevent and treat diabetic osteoporosis, fractures, and postmenopausal osteoporosis.

[0104] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following embodiments are generally carried out under conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and weight parts.

[0105] Example 1.

[0106] I. Study on mouse OVX osteoporosis model

[0107] 1.1 Method

[0108] As Figure 1 shown, it is the flow chart of OVX-induced osteoporosis and SRI37330 prevention experiment: Select adult (9-week-old) wild-type mice and divide them into a blank control group, a surgical control group, a low-concentration group and a high-concentration group, with 6 mice in each group, to establish an Ovx osteoporosis model. Intraperitoneal injection was continuously administered twice a week for 6 weeks after modeling (the blank control group was not administered, the surgical control group was administered phosphate buffer solution, the low-concentration group was administered at a concentration of 2 uM, and the high-concentration group was administered at a concentration of 10 uM). Calcein was intraperitoneally injected in the 4th week and alizarin red was intraperitoneally injected in the 5th week. Samples were taken at the 6th week after surgery for bone mass analysis of mouse tibia and histological staining of bone tissue. Bone tissue phenotypes were observed by Vonkussa staining, TRAP staining and double fluorescence labeling.

[0109] 1.2 Results

[0110] The results of the in vivo osteoporosis prevention experiment are as Figures 1-9 shown, among which: Figure 2 are the cancellous bone pictures of mouse tibia examined by microCT, which are the blank control group (sham operation group Sham), the surgical control group (OVX operation + Control / PBS), the low-concentration group (OVX + Low SRI-37330) and the high-concentration group (OVX + High SRI-37330) respectively. It can be seen from the figure that in terms of bone mass, the bone mass of the low-concentration group and the high-concentration group increased compared with that of the surgical control group. Especially, the high-concentration group was almost the same as the blank control group. It can be seen that the SRI-37330 treatment group (low concentration and high concentration) could partially rescue the bone loss caused by OVX, indicating that SRI-37330 has a preventive and therapeutic effect on osteoporosis in mice modeled by OVX.

[0111] Figure 3 is the statistical chart of cancellous bone of mouse tibia examined by microCT. The results show that: compared with the blank control group, the bone mass of the tibia in the surgical control group decreased significantly. The SRI-37330 treatment group (low concentration and high concentration) partially rescued the bone loss caused by OVX. The above results confirm that SRI-37330 has a preventive and therapeutic effect on osteoporosis in mice.

[0112] Figure 4 It is a picture of cancellous bone examined by Von kossa staining of mouse femur. As can be seen from the figure, on the longitudinal section of the femur (black is bone mass), the bone mass in the low-concentration group and the high-concentration group showed a certain degree of increase compared with the surgical control group, and the high-concentration group was slightly lower than the blank control group. It can be seen that the SRI-37330 treatment group (low concentration and high concentration) partially rescued the bone loss caused by OVX, indicating that SRI-37330 has a preventive and therapeutic effect on osteoporosis in OVX-induced mice. Figure 5 It is a statistical chart of cancellous bone examined by Von kossa staining of mouse femur. The results showed that compared with the blank control group, the bone mass of the tibia in the surgical control group was significantly reduced, and the SRI-37330 treatment group (low concentration and high concentration) partially rescued the bone loss caused by OVX. The above results confirmed that SRI-37330 has a preventive and therapeutic effect on osteoporosis in mice.

[0113] Figure 6 It is a picture of cancellous bone examined by double fluorescent labeling of mouse femur. As can be seen from the figure, the distance between the green fluorescence and the red fluorescence represents the bone formation rate within a certain period of time. Compared with the surgical control group, the bone formation rate in the treatment group (low concentration and high concentration) decreased slightly.

[0114] Figure 7 It is a statistical chart of the bone formation rate indicated by double fluorescent labeling. It can be seen that the SRI-37330 treatment group (low concentration and high concentration) further reduced the decrease in the bone formation rate caused by the control group. The bone formation rate in the SRI-37330 treatment group (low concentration and high concentration) decreased further compared with the control group.

[0115] Figure 8 It is a TRAP staining of mouse bone tissue sections. The results showed that the number of osteoclasts on the trabecular bone of mice in the blank control group increased; the number of osteoclasts on the trabecular bone of mice in the SRI-37330 treatment group (low concentration and high concentration) decreased compared with the control group.

[0116] Figure 9 It is a statistical chart of the TRAP staining results. It can be seen that the SRI-37330 treatment group (low concentration and high concentration) reversed the increase in the number of osteoclasts caused by the control group. The number of osteoclasts in the SRI-37330 treatment group (low concentration and high concentration) decreased compared with the control group. (* indicates P<0.05. ** indicates P<0.01. *** indicates P<0.001).

[0117] The above results confirmed that SRI-37330 treatment simultaneously inhibited osteoblasts and osteoclasts in vivo, promoting bone metabolism to enter low turnover to prevent and treat osteoporosis.

[0118] II. Research on mouse diabetic fracture model

[0119] 2.1 Method

[0120] Mice (4W) on a high - fat and high - sugar diet (HFD) for one month were intraperitoneally injected with a low dose (40 mg / kg) of streptozotocin (STZ) for one week and then continued to be fed under HFD conditions until 8W. The results of body weight and fasting plasma glucose (FPG) proved the successful establishment of a diabetic (FPG > 11.1 mmol / L) mouse model. As Figure 10 shown, 4 mice in each of the healthy control group (Control), diabetes group (Diabetes), diabetes + low - concentration treatment group (Diabetes + Low SRI), and high - concentration treatment group (Diabetes + High SRI) were used to establish a fracture model. Intraperitoneal injection was administered twice a week for 2 weeks after modeling, and then samples were taken for X - ray analysis of the bone mass of the mouse tibia to observe the fracture healing situation.

[0121] 2.2 Results

[0122] The experimental results of treating diabetic fractures in vivo are as Figure 10 shown:

[0123] Among them: The X - ray examination pictures of the mouse femur were the blank control group (sham - operation group Control), surgical control group (PBS), low - concentration group (Low SRI - 37330, concentration 2 μM), and high - concentration group (High SRI - 37330, concentration 10 μM). The results showed that compared with the blank control group, the fracture healing in the surgical control group was significantly limited, and the SRI - 37330 treatment groups (low - concentration and high - concentration) accelerated the fracture healing disorder caused by diabetes. The above results confirmed that SRI - 37330 has a preventive and therapeutic effect on diabetic fractures in mice.

[0124] Therefore, as a known research and pre - clinical TXNIP inhibitor, SRI - 37330 can not only reduce the secretion and action of glucagon and block hepatic glucose output, but also efficiently and specifically reduce the level of TXNIP protein in osteoblasts and osteoclasts, inhibit the differentiation of osteoblasts and osteoclasts, thereby having the function of promoting bone metabolism into low turnover to prevent and treat osteoporosis, fractures, and post - menopausal osteoporosis.

Claims

1. Use of a TXNIP-specific inhibitor in the preparation of a product for preventing and treating osteoporosis or fractures, characterized in that, The product is a composition or preparation containing SRI-37330, or its optical isomer, or its pharmaceutically acceptable salt; The TXNIP specific inhibitor is SRI-37330, and its chemical structural formula is as follows:

2. Use of a TXNIP-specific inhibitor according to claim 1 in the preparation of a product for preventing and treating osteoporosis or fractures, characterized in that, The said TXNIP specific inhibitor promotes bone metabolism into low turnover, including any one of the following: (i) Inhibiting the differentiation and absorption of osteoclasts, and the differentiation and mineralization of osteoblasts, thereby promoting the state of bone metabolism into low turnover; (ii) Promoting bone metabolism into low turnover to delay the occurrence of osteoporosis and promote the healing of diabetic fractures; (iii) Reducing the expression level of TXNIP in osteoblasts; (iv) Reducing the expression level of TXNIP in osteoclasts.

3. Use of a TXNIP specific inhibitor according to claim 1 in the preparation of a product for preventing and treating osteoporosis or fractures, characterized in that, The content of the TXNIP specific inhibitor SRI-37330 in the said composition or preparation is 0.001-99 wt%.

4. Use of a TXNIP specific inhibitor according to claim 1 in the preparation of a product for preventing and treating osteoporosis or fractures, characterized in that, The said composition is a pharmaceutical composition, a cosmetic composition, a dietary supplement, a food composition or a health care product composition.

5. Use of a TXNIP specific inhibitor according to claim 4 in the preparation of a product for preventing and treating osteoporosis or fractures, characterized in that, The said pharmaceutical composition contains (a) SRI-37330 or its pharmaceutically acceptable salt as an active ingredient; and (b) a pharmaceutically acceptable carrier or excipient.

6. Use of a TXNIP specific inhibitor according to claim 5 in the preparation of a product for preventing and treating osteoporosis or fractures, characterized in that, The said pharmaceutical composition further contains an active ingredient that additionally reduces glucagon secretion and action and blocks hepatic glucose output.

7. Use of a TXNIP specific inhibitor according to claim 4 in the preparation of a product for preventing and treating osteoporosis or fractures, characterized in that, The forms of the said pharmaceutical composition include: tablets, capsules, granules, suspensions, pills, solutions, syrups, or injections.

8. Use of a TXNIP-specific inhibitor according to claim 4 in the preparation of a product for preventing and treating osteoporosis or fractures, characterized in that, The said health care product composition contains SRI-37330 as an active ingredient for promoting bone metabolism into low turnover to prevent osteoporosis and promote fracture healing.

9. Use of a TXNIP-specific inhibitor according to claim 1 in the preparation of a product for preventing and treating osteoporosis or fractures, characterized in that, Adding the TXNIP specific inhibitor SRI-37330 or its salt to the cell culture system reduces the expression level of TXNIP protein in the said cells; or culturing cells in the presence of the TXNIP specific inhibitor SRI-37330 or its salt reduces the expression level of TXNIP protein in the said cells.

10. Use of a TXNIP-specific inhibitor according to claim 9 in the preparation of a product for preventing and treating osteoporosis or fractures, characterized in that, The concentration of the orally active TXNIP specific inhibitor in the cell culture system is 0.01-100 uM.

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