Application of bis-selenomaleimide in preparation of anti-osteoporosis drugs

Abstract

The invention relates to an application of a bis-selenomaleimide compound in preparation of an anti-osteoporosis medicine. The bis (seleno) maleimide compound is N-benzyl-3, 4-bis (phenylseleno)-maleimide (3k), and the structural formula of the bis (seleno) maleimide compound is shown in the description. Anti-osteoporosis activity evaluation is carried out on N-benzyl-3, 4-di (phenylseleno)-maleimide (3k), and it is found for the first time that N-benzyl-3, 4-di (phenylseleno)-maleimide (3k) has obvious anti-osteoporosis activity, osteoclast differentiation and bone resorption activity can be effectively inhibited, in-vivo bone loss can be inhibited, and osteoporosis can be inhibited. The compound can be used for preparing the anti-osteoporosis medicine, provides a novel anti-osteoporosis medicine for clinic, and has good clinical application value and wide application prospect.

Description

Technical Field

[0001] The invention relates to application of diselenomaleimide compounds in preparing anti-osteoporosis drugs, belonging to the technical field of anti-osteoporosis drugs. Background Art

[0002] Osteoporosis (OP) is a metabolic bone disease characterized by decreased bone mass and destruction of bone tissue microstructure, which leads to increased bone brittleness and easy fractures. It has a high mortality rate and creates an economic burden. In recent years, a series of studies have shown that excessive osteoclast production can cause excessive bone resorption, leading to unbalanced bone remodeling, bone loss, and osteoporosis. Therefore, using drugs to prevent osteoclast differentiation is considered to be one of the most effective strategies for treating such diseases. However, most existing drugs of this type are protein drugs, which have many defects such as serious adverse reactions, low efficacy, and complex preparation processes.

[0003] Selenium is one of the essential trace elements for the human body. Studies have shown that selenium plays an important role in regulating bone formation and bone metabolism. In recent years, the pharmaceutical activity of organic selenium compounds has been widely confirmed. However, there are currently no reports on the anti-osteoporosis activity of organic selenium compounds. Maleimide, also known as maleimide, exists as an active parent nucleus in many types of drugs and bioactive molecular structures. The anti-osteoporosis activity of maleimide compounds has been confirmed. Considering the effects of selenium and maleimide compounds on bones, and the important role of organic selenium compounds and maleimide compounds in the field of new drug research and development, and based on the current urgent need for osteoporosis treatment, designing and discovering new drugs for the treatment of osteoporosis has important clinical and practical significance. Summary of the invention

[0004] In view of the shortcomings of the prior art, the present invention provides the use of diselenomaleimide compounds in the preparation of anti-osteoporosis drugs. By studying the anti-osteoporosis activity of new diselenomaleimide compounds, new anti-osteoporosis active lead compounds are discovered, providing new ideas and experimental basis for the further development of drugs for the treatment of osteoporosis.

[0005] The technical solution of the present invention is as follows:

[0006] The invention provides application of a diselenomaleimide compound 3k in preparing an anti-osteoporosis drug.

[0007] The diselenomaleimide compound 3k is N-benzyl-3,4-di(phenylselenyl)-maleimide, and its structural formula is as follows:

[0008]

[0009] Preferably according to the present invention, the anti-osteoporosis drug includes a drug that inhibits bone loss or inhibits osteoclast differentiation.

[0010] Preferably according to the present invention, the anti-osteoporosis drug includes a drug that inhibits the formation of mature osteoclasts or inhibits the bone resorption activity of osteoclasts.

[0011] Preferably according to the present invention, the anti-osteoporosis drug includes a drug that inhibits the expression of osteoclast differentiation-related genes and proteins.

[0012] Preferably according to the present invention, the anti-osteoporosis drug inhibits osteoclast differentiation by inhibiting MAPK and NF-κB signaling pathways.

[0013] Preferably according to the present invention, the anti-osteoporosis drug is a pharmaceutical composition with the diselenomaleimide compound 3k as an active ingredient.

[0014] Further preferably, the anti-osteoporosis drug also includes a pharmaceutically acceptable excipient.

[0015] Preferably according to the present invention, the effective dose of the anti-osteoporosis drug is 5 mg / kg to 15 mg / kg every two days.

[0016] The beneficial effects of the present invention are:

[0017] The present invention provides the use of diselenomaleimide compounds in the preparation of anti-osteoporosis drugs. The present invention evaluates the anti-osteoporosis activity of N-benzyl-3,4-di(phenylselenyl)-maleimide (3k), and for the first time finds that N-benzyl-3,4-di(phenylselenyl)-maleimide (3k) has obvious anti-osteoporosis activity, can effectively inhibit osteoclast differentiation and its bone resorption activity, inhibit bone loss in the body, can be used to prepare anti-osteoporosis drugs, provides new anti-osteoporosis drugs for clinical use, and has good clinical application value and broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The results and cytotoxicity tests showed that 3k inhibited RANKL-induced osteoclast formation in a dose-dependent manner;

[0019] In the figure, A is a representative image of OC after BMMs were treated with gradient concentrations of 3k, scale bar = 500 μm; B is a quantitative analysis of the number of TRAP-stained multinucleated cells (≧3 nuclei) in each well (96-well plate); C is the effect of different concentrations of 3k on BMMs measured by MTT method.

[0020] Figure 2 The result is that 3k inhibits osteoclast formation and its bone resorption activity;

[0021] In the figure, A is a representative confocal image of OC stained for F-actin and nuclei, scale bar = 200 μm; B is the average number of nuclei per unit cell; C is the average number of osteoclasts per unit area; D is a representative image of the eroded area on the hydroxyapatite-coated plate, scale bar = 200 μm; E is the percentage of osteoclast resorption area.

[0022] Figure 3 3k inhibits the expression of osteoclast-specific genes and proteins;

[0023] In the figure, A is the expression level of the specific gene TRAP; B is the expression level of the specific gene C-Fos; C is the expression level of the specific gene NFATc1; D is the expression level of the specific gene MMP-9; E is the expression level of the specific gene Cathepsin K; F is the expression level of the specific gene DC-STAMP; G is the expression levels of C-Fos, NFATc1, MMP-9 and Cathepsin K in BMMs in the 3k-treated group and the control group, respectively; H is a statistical graph of the difference in C-Fos protein expression level between the 3k-treated group and the control group; I is a statistical graph of the difference in NFATc1 protein expression level between the 3k-treated group and the control group; J is a statistical graph of the difference in MMP-9 protein expression level between the 3k-treated group and the control group; K is a statistical graph of the difference in Cathepsin K protein expression level between the 3k-treated group and the control group.

[0024] Figure 4 The results showed that 3k inhibited RANKL-induced MAPK and NF-κB signaling pathways;

[0025] In the figure, A is the expression level of pp38, p38, P-ERK, ERK, P-JNK, and JNK in BMMs of 3k treatment group and control group respectively; B is the quantitative determination of the relative ratio of phosphorylated p38 protein to unphosphorylated p38 protein; C is the quantitative determination of the relative ratio of phosphorylated ERK protein to unphosphorylated ERK protein; D is the quantitative determination of the relative ratio of phosphorylated JNK protein to unphosphorylated JNK protein; E is the expression level of pp65, p65, p-IκBα, and IκBα in BMMs of 3k treatment group and control group respectively; F is the quantitative determination of the relative ratio of phosphorylated p65 protein to unphosphorylated p65 protein; G is the quantitative determination of the relative ratio of phosphorylated IκBα protein to unphosphorylated IκBα protein.

[0026] Figure 5 Improvement of OVX-induced bone loss results for 3k;

[0027] In the figure, A is the femoral structure captured by high-resolution micro-CT and processed by 3D computer reconstruction; B is the quantitative measurement of the relative bone volume (BV / TV) in the three groups of Sham, OVX and OVX+3k (10 mg / Kg); C is the quantitative measurement of the trabecular thickness (Tb.Th) in the three groups of Sham, OVX and OVX+3k (10 mg / Kg); D is the quantitative measurement of the number of trabeculae (Tb.N) in the three groups of Sham, OVX and OVX+3k (10 mg / Kg); E is the quantitative measurement of the trabecular separation (Tb.sp) in the three groups of Sham, OVX and OVX+3k (10 mg / Kg). DETAILED DESCRIPTION

[0028] In order to better understand the essence of the present invention, the present invention is further described below in conjunction with the embodiments, but they should not be regarded as limiting the present invention.

[0029] The N-benzyl-3,4-di(phenylselenyl)-maleimide (3k) described in the present invention is prepared according to the method in the document C-H Selenation and Monoselenation of Electron-Deficient Alkenes via Radical Coupling at Room Temperature.

[0030] Example 1: Diselenomaleimide compound 3k inhibits RANKL-induced osteoclast formation in a dose-dependent manner

[0031] 1. Cell culture of mouse bone marrow-derived macrophages (BMMs): After 6-8 week-old wild-type (WT) mice were killed, the femurs and tibias were dissected out; after washing with sterile PBS buffer added with gentamicin, the epiphyseal ends of the bones were cut off, and α-MEM culture medium was drawn with a 5 mL syringe and inserted into the bone marrow cavity. The bone marrow was blown out until the bone tissue turned white, and the flushed cell suspension was collected into a culture bottle and placed in a constant temperature incubator at 37°C and 5% CO2 for 3 days. The adherent cells visible under an inverted microscope are mouse bone marrow-derived macrophages (BMMs).

[0032] 2. Prepare osteoclast induction solution: Add 1 mL of PBS buffer to the recombinant mouse RANKL protein lyophilized powder to prepare a 10 μg / mL RANKL protein solution as the storage concentration and store at -20°C. Take out the freeze-thaw before use (freeze-thaw no more than 3 times) and prepare α-MEM complete medium with a RANKL protein concentration of 50 ng / mL, which is the osteoclast induction solution.

[0033] 3. Diselenomaleimide compound 3k inhibits RANKL-induced osteoclast formation in a dose-dependent manner: BMMs cells were cultured at a rate of 1×10 4 The cells were seeded in a 96-well plate at a density of 100 / well. After 24 hours, osteoclast induction solution and diselenomaleimide compound 3k were added respectively. The cells were divided into control group, RANKL group and RANKL+3k group. The gradient concentration of 3k in RANKL+3k group was 0, 0.0625, 0.125, 0.25, 0.5, 1, 2 μM. The control group was added with the same volume of PBS buffer. The medium was changed once every 3 days. After round-shaped osteoclasts were observed under the microscope, TRAP staining was performed. The osteoclast induction solution was discarded, the cells were washed 3 times with PBS buffer, and 4 % paraformaldehyde was fixed at 37°C in a light-proof environment for 20 minutes, and then washed with PBS buffer for 3 times; the staining solution was then placed in a 37°C water bath for preheating for 5 minutes to ensure that the TRAP staining solution was at 37°C, and at the same time, the ultrapure water at 37°C in the well plate was replaced to keep the well plate warm at 37°C; 100 μL of staining solution was added to each well of the 96-well plate, and incubated in a light-proof 37°C constant temperature box for 2 hours, then the staining solution was aspirated and discarded, and the cells were washed with PBS buffer for 3 times. The images were observed and taken using a stereomicroscope, and the number of osteoclasts was counted. Cells with more than or equal to 3 nuclei were osteoclasts. The results were as follows Figure 1 AB. Figure 1 AB shows that diselenomaleimide compound 3k can effectively inhibit osteoclast formation, and the inhibitory ability is related to the concentration of diselenomaleimide compound 3k, which is a dose-dependent inhibition. As the concentration of diselenomaleimide compound 3k increases, the ability to inhibit osteoclast formation increases.

[0034] 4. Cytotoxicity test: BMMs cells were cultured at 1×10 4 The cells were inoculated in a 96-well plate at a density of 100 μL / well. After 24 hours, osteoclast induction solution and diselenomaleimide compound 3k were added respectively, and the cells were divided into a control group and a drug group 3k. The gradient concentration of 3k in the RANKL+3k group was 0, 0.0625, 0.125, 0.25, 0.5, 1, 2, 5, 10, and 20 μM. The control group was added with the same volume of PBS buffer, which was replaced with ordinary complete culture medium after 48 hours. Then, MTT was added at 10 μL / well, and the cells were incubated at 37°C, 5% CO2 in a constant temperature and away from light for 4 hours. The old liquid was sucked away by a liquid suction vacuum pump, and DMSO was added at 100 μL / well. The cells were shaken on a horizontal shaker in a dark place for 10 minutes. The OD value at 490 nm was detected by a multifunctional microplate reader. The experiment was repeated 3 times. The results are as follows Figure 1 As shown in C.

[0035] Depend on Figure 1 C shows that the diselenomaleimide compound 3k has basically no obvious cytotoxicity at the tested concentration.

[0036] Example 2: Diselenomaleimide compound 3k inhibits osteoclast formation and bone resorption activity

[0037] 1. Diselenomaleimide compound 3k inhibits osteoclast formation: BMMs cells were plated at a density of 8000 cells / well in a 96-well plate filled with normal complete culture medium (containing 50 ng / mL RANKL protein and 30 ng / mL M-CSF). After 24 hours, diselenomaleimide compound 3k with gradient concentrations (0, 0.5, 1 and 2 μM) were added respectively as the compound group, and BMMs cells were cultured in normal complete culture medium as the control group (without adding diselenomaleimide compound 3k). The culture medium was changed every 2 days. After the 5th day, an obvious actin ring was observed under the mirror image, and then the original culture medium was discarded, washed with PBS buffer for 3 times, then fixed with 4% paraformaldehyde for 15 minutes, washed twice with PBS buffer, then permeabilized with 5% TritonX-100 for 20 minutes, and finally washed 3 times with PBS buffer. Pour away the PBS buffer, then add rhodamine-labeled phalloidin for staining, incubate in the dark for 30 minutes, then wash twice with PBS buffer; finally add DAPI staining solution, let stand for 10 minutes, wash twice with PBS, add anti-fluorescence sealing solution; finally, observe the osteoclast actin ring under an inverted fluorescence microscope, and calculate its number and volume. The results are as follows Figure 2 As shown in A~C.

[0038] Depend on Figure 2 As shown in A to C, compared with the control group, the actin ring in the osteoclasts in the compound group was significantly reduced, indicating that the diselenomaleimide compound 3k can effectively inhibit the formation of mature osteoclasts.

[0039] 2. Diselenomaleimide compound 3k inhibits osteoclast resorption activity: 48-well hydroxyapatite bone plates were soaked in complete culture medium for 24 h, and then BMMs cells were cultured at a rate of 1×10 4 The cells were plated at a density of 100 μM / well in a bone plate filled with normal complete culture medium (containing 50 ng / mL RANKL protein and 30 ng / mL M-CSF). After 24 hours, diselenomaleimide compound 3k with gradient concentrations (0, 0.5, 1 and 2 μM) were added as the compound group, and BMMs cells were cultured in normal complete culture medium as the control group (without adding diselenomaleimide compound 3k). The medium was changed every 2 days (careful operation should be performed when changing the medium to avoid damaging the cells). After the 5th day, pits could be observed in the hydroxyapatite bone plate under the microscope, and then the cells on the bone slices were removed with 5% sodium hypochlorite solution, and then the 5% sodium hypochlorite was washed off with 75% alcohol. After drying, the cells were observed and photographed under an inverted microscope, and the absorption area of ​​the hydroxyapatite bone plate in each well was calculated and statistically analyzed. The results are shown in FIG. Figure 2 As shown in D~E.

[0040] Depend on Figure 2 As shown in D-E, compared with the control group, the absorbed area on the bone slices in the compound group was significantly reduced, indicating that the diselenomaleimide compound 3k can effectively inhibit the absorption activity of osteoclasts.

[0041] Example 3: Diselenomaleimide compound 3k inhibits the expression of osteoclast-specific genes and proteins

[0042] BMMs cells were cultured with RANKL protein (RANKL group) and diselenomaleimide compound 3k (gradient concentration 0.5, 1, 2 μM, 3k treatment group) for 5 days. At the same time, PBS buffer was used instead of RANKL protein and diselenomaleimide compound 3k for co-culture as the control group. After the samples were collected, qRT-PCR and Western Blotting were performed to detect the mRNA and protein expression of key genes in osteoclasts. The results are shown in Figure 2 Figure 3 As shown in A to K. The mRNA and protein of the key osteoclast genes include TNFR receptor associated factor (TRAP), FBJ osteosarcoma oncogene (C-Fos), nuclear factor of activated T cells 1 (NFATc1), matrix metalloproteinase 9 (MMP-9), cathepsin k (Cathepsin K) and dendritic cell specific transmembrane protein (DC-STAMP).

[0043] Depend on Figure 3 A-K show that compared with the control group, the expression levels of key genes and proteins of osteoclasts in the RANKL group were significantly increased. After intervention with the diselenomaleimide compound 3k, the expression levels of key genes and proteins of osteoclasts in the 3k treatment group were significantly downregulated compared with the RANKL group, indicating that the diselenomaleimide compound 3k can effectively inhibit the expression of key genes and proteins of osteoclasts, thereby inhibiting osteoclast differentiation and its bone resorption activity.

[0044] Example 4: Diselenomaleimide compound 3k inhibits RANKL-induced MAPK and NF-κB signaling pathways

[0045] RANKL protein binds to its receptor RANK and can activate downstream MAPK and NF-κB signaling pathways, thereby promoting osteoclast differentiation. In order to investigate the mechanism of action of diselenomaleimide compound 3k in inhibiting osteoclast differentiation, this example investigates the effect of diselenomaleimide compound 3k on RANKL-induced MAPK and NF-κB signaling pathways.

[0046] 1. BMMs cells were cultured with RANKL protein (RANKL group) and diselenomaleimide compound 3k (1 μM, 3k treatment group) for 0, 5, 10, 20, 30, and 60 min, and Western Blotting was performed to detect the expression of related proteins. The results are shown in Figure 2. Figure 4 As shown in A to D. The related proteins include phosphorylated p38 mitogen-activated protein kinase (pp38), p38 mitogen-activated protein kinase (p38), phosphorylated extracellular signal-regulated protein kinase (p-ERK), extracellular signal-regulated protein kinase (ERK), phosphorylated c-Jun amino-terminal kinase (p-JNK) and c-Jun amino-terminal kinase (JNK)).

[0047] Depend on Figure 4 A to D show that when BMMs cells are stimulated by RANKL protein, the phosphorylation levels of p38, ERK and JNK all tend to increase first and then decrease. However, when cells are simultaneously exposed to RANKL protein and diselenomaleimide compound 3k, the phosphorylation levels of p38, ERK and JNK are significantly inhibited. Among them, the phosphorylation levels of p38 and ERK are more obviously inhibited after 10min and 20min of simultaneous stimulation of RANKL protein and diselenomaleimide compound 3k, while the phosphorylation level of JNK is more obvious at 20min and 30min of stimulation. The above results all indicate that diselenomaleimide compound 3k can effectively inhibit the MAPK signaling pathway induced by RANKL protein.

[0048] 2. BMMs cells were cultured with RANKL protein (RANKL group) and diselenomaleimide compound 3k (1 μM, 3k treatment group) for 0, 5, 10, 20, 30, and 60 min, respectively. After the samples were collected, BMMs cells were treated with RANKL and 3k (1 μM) and then subjected to Western Blotting to detect the expression of related proteins. The results are shown in Figure 2. Figure 4 As shown in E to G. The related proteins include phosphorylated transcription factor p65 (pp65), transcription factor p65 (p65), phosphorylated nuclear factor κB inhibitor protein (p-IκBα) and nuclear factor κB inhibitor protein (IκBα).

[0049] Depend on Figure 4As shown in E to G, when BMMs cells were stimulated by RANKL protein, the phosphorylation levels of p65 and IκBα increased first and then decreased. However, when cells were simultaneously acted upon by RANKL protein and compound 3k, the phosphorylation levels of p65 and IκBα were significantly inhibited. The above results indicate that diselenomaleimide compound 3k can effectively inhibit the NF-κB signaling pathway induced by RANKL protein.

[0050] Example 5: Diselenomaleimide compound 3k can improve OVX-induced bone loss

[0051] In order to further evaluate the anti-osteoporosis activity of the diselenomaleimide compound 3k, this example further investigated the inhibitory effect of the diselenomaleimide compound 3k on bone loss in osteoporotic mice.

[0052] 1. Experimental animals: 30 6-week-old SPF-grade non-pregnant healthy female C57BL / 6J mice, weighing about 25g, were raised in the Animal Experiment Center of Wenzhou Medical University, with a constant temperature (20-24℃), 12-hour day and night light cycle, free drinking water and food, and litter replacement and feed and water addition were all carried out in an independent ventilation system. The treatment of animals complies with relevant animal ethics requirements.

[0053] 2. Establishment of osteoporosis mouse model (OVX): The model was established after mice were adaptively fed for one week. The specific method is as follows:

[0054] 1) Anesthesia: After weighing the mouse, calculate the corresponding anesthetic dose, draw a mixed anesthetic of ketamine and xylazine with a syringe, and inject it after aspirating blood from the mouse's abdominal cavity;

[0055] 2) Skin preparation: Find the intersection of the lower rib margin and the spine on both sides of the mouse's back, and prepare the skin in an area with a diameter of about 2 cm with this point as the center;

[0056] 3) Surgery: A 0.5 cm long incision was made at a distance of 0.5 cm from the spine. The mouse fat tissue and ovaries were found according to the physiological structure of the mouse. The fallopian tubes were first ligated with surgical sutures, and then the ovaries were removed. Strict sutures were performed layer by layer from deep to shallow to obtain an osteoporosis mouse model (OVX). The same operation was performed on the contralateral side. The sham operation group mice (Sham) underwent the same steps as above, but when the ovaries were found, the fat tissue near the ovaries was cut off.

[0057] 4) Inject an appropriate dose of penicillin intramuscularly to prevent infection.

[0058] 3. Experimental grouping: Thirty 6-week-old SPF-grade non-pregnant healthy female C57BL / 6 mice were randomly divided into 3 groups, with 10 mice in each group. The 3 groups were named as: sham operation group (Sham+Vehicle), solvent group (OVX+Vehicle) and drug group (OVX+drug).

[0059] 4. Administration method: First dissolve the diselenomaleimide compound 3k in DMSO to obtain a diselenomaleimide compound 3k solution; add physiological saline to dilute DMSO to reduce its concentration to a safe range to obtain a DMSO solution. The sham operation group and the solvent group were gavaged with 0.2 mL of DMSO solution, and the drug group was gavaged with 10 mg / kg diselenomaleimide compound 3k solution every other day for 8 consecutive weeks. Then, micro-CT detection was performed to qualitatively and quantitatively analyze the bone parameters that reflect the role of maintaining bone mass. The results are as follows: Figure 5 As shown in A to E.

[0060] Depend on Figure 5 A shows that after the ovaries were removed, the bone mass of mice decreased significantly, and the osteoporosis mouse model (OVX) was successfully established. The bone mass of the mice in the solvent group was significantly lower than that in the sham operation group, while the bone mass of the OVX mice treated with the diselenomaleimide compound 3k increased compared with the solvent group. The reconstructed 3D images more intuitively illustrate the changes in bone mass.

[0061] Depend on Figure 5 B-E show that compared with the solvent group, the femur-related parameters including relative bone volume (BV / TV), trabecular number (Tb.N), and trabecular thickness (Tb.Th) in the sham group were significantly decreased; compared with OVX, after treatment with diselenomaleimide 3k, bone volume (BV / TV), trabecular number (Tb.N), and trabecular thickness (Tb.Th) were significantly increased, and trabecular separation (Tb.sp) was significantly reduced. This indicates that diselenomaleimide 3k has a protective effect on bone loss in mice caused by ovariectomy.

[0062] In summary, the present invention evaluated the anti-osteoporosis activity of N-benzyl-3,4-di(phenylselenyl)-maleimide (3k), and found for the first time that N-benzyl-3,4-di(phenylselenyl)-maleimide (3k) has obvious anti-osteoporosis activity, can effectively inhibit osteoclast differentiation and its bone resorption activity, inhibit bone loss in vivo, can be used to prepare anti-osteoporosis drugs, and provides new anti-osteoporosis drugs for clinical use, which has good clinical application value and broad application prospects.

Claims

1. Application of diselenomaleimide compound 3k in the preparation of anti-osteoporosis drugs, characterized in that: The diselenomaleimide compound 3k is N-benzyl-3,4-di(phenylselenyl)-maleimide, and its structural formula is as follows:

2. The use according to claim 1, characterized in that The anti-osteoporosis drugs include drugs that inhibit bone loss or inhibit osteoclast differentiation.

3. The use according to claim 1, characterized in that The anti-osteoporosis drugs include drugs that inhibit the formation of mature osteoclasts and inhibit the bone resorption activity of osteoclasts.

4. The use according to claim 1, characterized in that The anti-osteoporosis drugs include drugs that inhibit the expression of osteoclast differentiation-related genes and proteins.

5. The use according to claim 1, characterized in that The anti-osteoporosis drug inhibits osteoclast differentiation by inhibiting MAPK and NF-κB signaling pathways.

6. The use according to claim 1, characterized in that The anti-osteoporosis drug is a pharmaceutical composition with diselenomaleimide compound 3k as an active ingredient.

7. The use according to claim 6, characterized in that The anti-osteoporosis drug also includes pharmaceutically acceptable excipients.

8. The use according to claim 7, characterized in that The effective dose of the anti-osteoporosis drug is 5 mg / kg to 15 mg / kg every two days.

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