Application of a bile acid in the preparation of a drug for treating osteoporosis

Abstract

This invention relates to the pharmaceutical field, specifically disclosing the application of bile acids in the preparation of drugs for treating osteoporosis. Studies have found that the content of porcine deoxycholic acid (HDCA) is significantly reduced in older individuals, and that HDCA supplementation can improve bone mineral density and bone microstructure. Animal experiments show that oral gavage of HDCA can significantly inhibit bone loss and improve bone quality, with its effect dependent on the bile acid receptor TGR5. This invention is the first to propose HDCA as an active ingredient for treating osteoporosis, providing a new drug candidate and mechanism of action for the prevention and treatment of osteoporosis.

Description

[0001] Cross-references

[0002] This application claims priority to Chinese Patent Application No. 2024113497425 (filed on September 26, 2024), the entire contents of which are incorporated herein by reference. Technical Field

[0003] This invention relates to the field of pharmaceuticals, and more particularly to the use of a bile acid in the preparation of a medicament for treating osteoporosis. Background Technology

[0004] Osteoporosis is a manifestation of aging in the skeletal system. It is primarily caused by bone loss and reduction, damage to bone microstructure, and increased bone fragility, leading to a high risk of fractures. Currently, osteoporosis has become a significant health problem for people over 50 years of age in my country.

[0005] Studying the pathogenesis of osteoporosis is beneficial for finding new therapeutic targets and for the diagnosis and treatment of the disease. However, the pathogenesis of osteoporosis is complex and influenced by many factors. The bone niche environment is rich in cell types and has close communication. At the same time, although significant progress has been made in the treatment of osteoporosis, poor patient compliance and adverse reactions to drug treatment still exist.

[0006] Therefore, studying the pathophysiological process of osteoporosis and finding potential new treatment options for osteoporosis remains an urgent task. Summary of the Invention

[0007] Based on the above analysis, the present invention aims to provide the application of bile acids in the preparation of anti-osteoporosis drugs, in order to solve the technical problems of osteoporosis prevention and treatment and the lack of early intervention drugs in the prior art.

[0008] The objective of this invention is mainly achieved through the following technical solutions:

[0009] Given the limited availability of osteoporosis prevention and treatment drugs in the existing technology, as well as their side effects and poor compliance, the purpose of this invention is to provide an application of bile acids in the preparation of osteoporosis treatment drugs.

[0010] To achieve the above objectives, this invention, through metabolomics and animal experiments, demonstrated that porcine deoxycholic acid (HDCA) can serve as an effective active ingredient for the prevention and treatment of osteoporosis. Specifically, HDCA levels are significantly reduced in aged mice, and HDCA supplementation can improve bone microstructure and increase bone mineral density. Further research revealed that this effect depends on the presence of the bile acid receptor TGR5, suggesting a clear mechanism of action.

[0011] The technical solution provided by this invention is to apply porcine deoxycholic acid to the preparation of a drug for treating osteoporosis. The drug can be prepared into oral formulations such as tablets and capsules, and can be supplemented with conventional pharmaceutical excipients. This invention's drug can inhibit bone loss and improve bone quality through oral administration.

[0012] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0013] (1) It was the first time that HDCA was found to significantly prevent and treat osteoporosis in animal models;

[0014] (2) HDCA is proposed as an active pharmaceutical ingredient, providing a new option for the treatment of osteoporosis;

[0015] (3) Clarify that its mechanism of action depends on the TGR5 receptor, providing a theoretical basis for subsequent drug development. Attached Figure Description

[0016] The accompanying drawings are for illustrative purposes only and are not intended to limit the invention. Throughout the drawings, the same reference numerals denote the same parts.

[0017] Figure 1 This invention utilizes serum samples from aged mice (18-month-old C57 / BL6 mice) and young mice (2-month-old C57 / BL6 mice) to perform bile acid-targeted metabolomics detection and evaluate their HDCA content.

[0018] Figure 2 This invention utilizes micro-CT scans of the fifth lumbar vertebra (L5) from aged mice (18-month-old C57 / BL6 mice) and young mice (2-month-old C57 / BL6 mice) to assess their bone microstructure.

[0019] Figure 3 The data represents the correlation analysis between HDCA abundance and bone volume fraction of the fifth lumbar vertebra, detected in aged mice (18-month-old C57 / BL6 mice) and young mice (2-month-old C57 / BL6 mice).

[0020] Figure 4 shows a comparison of the effects of gavage administration of saline and HDC on aged mice (18-month-old C57 / BL6 mice). Figure 4a For dynamic bone mineral density results, Figure 4b This is the result of a micro-CT scan.

[0021] Figure 5 shows a comparison of HDCA administered by gavage to wild-type mice (2-month-old C57 / BL6 mice) and TGR5 knockout mice (2-month-old mice). Figure 5a For dynamic bone mineral density results, Figure 5bThis is the result of a micro-CT scan. Detailed Implementation

[0022] The preferred embodiments of the present invention will now be described in detail with reference to the accompanying drawings, which constitute a part of the present invention and are used together with the embodiments of the present invention to illustrate the principles of the present invention, but are not intended to limit the scope of the present invention.

[0023] It should be noted that in the intestines, through enzymatic reactions by intestinal microorganisms, bile acids undergo further metabolism to become secondary bile acids, and porcine deoxycholic acid is one such secondary bile acid. Experiments have shown that porcine deoxycholic acid (… Hyodesoxycholic Acid, HDCA The expression of this substance was lower in older mice than in younger mice, and both were strongly correlated with bone quality.

[0024] It should be noted that bile acids can be used as an indicator of bone quality, and furthermore, porcine deoxycholic acid, a type of bile acid, can be used as an indicator of bone quality.

[0025] In this invention, porcine deoxycholic acid is collected from blood samples of research subjects (subjects), and the relative abundance of porcine deoxycholic acid and bone mineral density are used to predict the bone mass of the research subjects.

[0026] This invention obtains the bone quality of research subjects by comparing the bone mass predicted using porcine deoxycholic acid with the normal bone mass. If the actual measured bone density is higher than the normal bone density (or the theoretically calculated bone density), it is considered normal; if the actual measured bone density is lower than the normal bone density, there is a risk of osteoporosis.

[0027] For example, a patient's bile acid metabolism profile sequencing (i.e., the blood test for bile acids in this invention) revealed a relatively low abundance of porcine deoxycholic acid (HDCA) of 0.01%, indicating a high risk of osteoporosis. A DXA examination showed a bone mineral density (BMD) of 0.463 g / cm² in the femoral neck and a T-score of -3.5, confirming the diagnosis of osteoporosis and demonstrating successful prediction.

[0028] In addition, the relative abundance of porcine deoxycholic acid and bone mineral density were correlated to predict the researchers' bone mass by establishing a bone mass prediction model.

[0029] The process of establishing the above bone mass prediction model includes the following steps:

[0030] Step 1: Data preparation;

[0031] Data on the relative abundance of porcine deoxycholic acid and bone mineral density from researchers were collected and cleaned, including handling missing, outlier, and duplicate values.

[0032] Step 2: Recruit a large number of healthy subjects (100 people) and, with the support of large sample data, perform a correlation analysis between the relative abundance of porcine deoxycholic acid and bone mineral density. The Pearson correlation analysis method was selected as the correlation analysis method.

[0033] The correlation coefficient was calculated using the Pearson correlation analysis method. The value is between -1 and 1. A value close to 1 indicates a strong positive correlation, a value close to -1 indicates a strong negative correlation, and a value close to 0 indicates no obvious linear relationship.

[0034] Step 3: Interpretation of Results;

[0035] Explain the relationship between variables based on the value and sign of the correlation coefficient.

[0036] Step 4: Significance test;

[0037] Perform a statistical significance test to determine whether the correlation is statistically significant.

[0038] Step 5: Data visualization;

[0039] Using scatter plots to show the relationships between variables helps to intuitively understand the correlations.

[0040] Step 6, Application.

[0041] The bone mass prediction model was validated, including verifying its accuracy and reliability on independent sample sets.

[0042] The relative abundance of porcine deoxycholic acid is positively correlated with bone mineral density.

[0043] On the other hand, the present invention also provides an application of bile acids as markers of bone metabolism, the bile acids including porcine deoxycholic acid.

[0044] It should be noted that the process of predicting bone mass using bile acids in this invention includes the following steps:

[0045] S1. Identify the research subjects;

[0046] Select target populations, such as postmenopausal women, older adults, or patients at known risk of osteoporosis.

[0047] S2. Collect blood samples;

[0048] It should be noted that bile acids are a type of substance used to measure the level of bile acids in serum or plasma by collecting blood samples from research subjects.

[0049] S3. Determine the bile acid content in the sample;

[0050] The bile acid content in blood samples can be accurately determined using enzymatic methods or liquid chromatography-mass spectrometry (LC-MS).

[0051] In step S3 above, the process of detecting bile acids using LC-MS is as follows:

[0052] S31. Sample preparation;

[0053] Collect blood samples and perform appropriate pretreatments, such as protein precipitation, liquid-liquid extraction, or solid-phase extraction (SPE), to enrich bile acids and remove impurities.

[0054] S32. Optimization of chromatographic conditions;

[0055] Choose a suitable chromatographic column (such as a C18 column) and mobile phase (such as a methanol-water mixture), and optimize gradient elution conditions to achieve efficient separation of bile acids.

[0056] S33. Mass spectrometry conditions optimization;

[0057] Set mass spectrometry parameters, such as the operating mode of the electrospray ionization source (ESI) (positive or negative ion mode) and the monitoring ion pairs in multiple reaction monitoring (MRM) mode.

[0058] S34. Preparation of standard curve;

[0059] A series of bile acid standard solutions with known concentrations were prepared to establish a standard curve, thereby enabling quantitative analysis of bile acid concentrations in samples.

[0060] S35. Sample Analysis: Inject the processed sample into a liquid chromatography system for chromatography-mass spectrometry analysis.

[0061] S36. Data Analysis: By comparing the chromatographic peaks in the sample with those in the standard, and combining the mass spectrometry data, qualitative and quantitative analysis of bile acids is performed.

[0062] Qualitative analysis revealed that the bile acids included porcine deoxycholic acid; quantitative analysis showed that the concentration range of porcine deoxycholic acid in the bile acids was 1.927–19.266 μg.

[0063] S37. Result verification;

[0064] The effectiveness of the method was verified through recovery experiments, precision and accuracy tests.

[0065] S4. Measure the subject's bone mass;

[0066] The bone quality detection methods of the present invention include single photon absorptiometry (SPA), dual-energy X-ray absorptiometry (DXA), quantitative computed tomography (QCT), and quantitative ultrasound measurement. All of these bone quality detection methods can obtain bone quality and then evaluate it.

[0067] It should be noted that the DXA detection process of the present invention includes the following steps:

[0068] S1. Patient preparation;

[0069] The patient needs to remove any metal objects that may interfere with the measurement.

[0070] S2, Positioning;

[0071] The patient is placed on the DXA scanner examination table, usually in a supine position.

[0072] S3, Scan;

[0073] The scanner emits X-rays, and the computer analyzes the attenuation of the X-rays.

[0074] S4. Results Analysis;

[0075] The software analyzes the data and generates reports including BMD, T-value, and Z-value.

[0076] S5. Report Interpretation;

[0077] Based on the results of the DXA report and the patient's clinical condition, the doctor will make a diagnosis and treatment recommendation.

[0078] General steps of QCT testing:

[0079] S1. Patient preparation;

[0080] The patient needs to remove any metal objects that may interfere with the scan and assume an appropriate position as instructed by the technician, usually in a supine position.

[0081] S2, positioning and scanning;

[0082] The patient is placed in a CT scanner to ensure that the area to be scanned (such as the lumbar spine or hip) is within the scanning field. A calibration phantom may be used during a CT scan to ensure the accuracy of the images.

[0083] S3. Image Acquisition: Use a CT scanner to obtain cross-sectional images of the patient.

[0084] S4. Image Processing: The scanned images need to be processed by specialized QCT analysis software. This typically includes image calibration, segmentation, and determination of the region of interest (ROI).

[0085] S5. Data Analysis: The software analyzes the images and calculates bone density, usually expressed in mg / cm³. QCT can measure the density of cortical bone and trabecular bone separately.

[0086] S6. Result Interpretation: Evaluate the patient's bone density based on the QCT results and compare it with normal values or the bone density of the same-aged population.

[0087] S7. Report Writing: Write a report based on the analysis results, which may include bone density values, T-score, Z-score, and an assessment of the fracture risk.

[0088] S8. Follow-up Guidance: Based on the QCT results, the doctor may recommend specific lifestyle changes, drug treatments, or other interventions for the patient.

[0089] The evaluation of bone quality in the present invention includes judging bone mass, degree of bone loss, and osteoporosis risk assessment.

[0090] Bone Mass: Taking DXA as an example, the T value is measured by DXA. T value: The T value is calculated by comparing the bone density measured in the patient with a normal reference value and standard deviation (SD). Normal bone mass: T value ≥ -1.0; Low bone mass: -2.5 < T value < -1.0; Osteoporosis: T value ≤ -2.5; After the patient undergoes DXA examination, the result shows that the bone mineral density BMD of the femoral neck is 0.463 g / cm 2 , the T value is -3.5, and the diagnosis is osteoporosis. Taking QCT as an example, after the patient undergoes QCT examination, the result shows that the lumbar spine bone density is 70 mg / cm 3 , and the diagnosis is osteoporosis.

[0091] Degree of Bone Loss: Taking DXA as an example, after the patient undergoes the first DXA examination, the result shows that the bone mineral density BMD of the femoral neck is 0.463 g / cm 2 , the T value is -3.5, and the diagnosis is osteoporosis. For the second DXA with added memory, the result shows that the bone mineral density BMD of the femoral neck is 0.784 g / cm2, the T value is -1.9, and the diagnosis is osteopenia, and bone loss is inhibited (bone mass recovery).

[0092] Fracture Risk Assessment: Combine the FRAX score to assess the fracture risk of the patient.

[0093] S4. Data Analysis;

[0094] The Pearson correlation coefficient method was used to analyze the correlation between bile acid levels and bone mineral density.

[0095] Calculate the Pearson correlation coefficient (r) using the following formula:

[0096] (1)

[0097] Where n is the sample size, and x and y are the values ​​of the variables. This indicates a summation.

[0098] S5. Establish a bone mass prediction model;

[0099] Based on the correlation analysis between bile acid levels and bone mineral density, a mathematical model was established to predict an individual's bone mineral density.

[0100] In step S5, the correlation analysis between bile acid levels and bone mineral density includes the following steps:

[0101] S51. Data preparation;

[0102] Collect relevant data and clean it, including handling missing values, outliers, and duplicate values.

[0103] The aforementioned data refers to the relative abundance of porcine deoxycholic acid obtained from individual gut microbiota sequencing obtained from S36 and the results derived from bone mineral density.

[0104] S52. Recruit a large number of healthy individuals to expand the sample size and perform the aforementioned testing and statistical analysis. With the support of this large sample data, conduct a correlation analysis between the relative abundance of porcine deoxycholic acid and bone mineral density.

[0105] S53. Select Pearson correlation analysis as the correlation analysis method;

[0106] The correlation coefficient is calculated using the formula in the Pearson correlation analysis method. The value is between -1 and 1. A value close to 1 indicates a strong positive correlation, a value close to -1 indicates a strong negative correlation, and a value close to 0 indicates no obvious linear relationship.

[0107] S54. Interpretation of results;

[0108] Explain the relationship between variables based on the value and sign of the correlation coefficient.

[0109] S55, Significance test;

[0110] Perform a statistical significance test to determine whether the correlation is statistically significant.

[0111] S56, Data Visualization;

[0112] Using scatter plots to show the relationships between variables helps to intuitively understand the correlations.

[0113] S57, Application;

[0114] S6. Validate the bone mass prediction model;

[0115] The accuracy and reliability of the bone mass prediction model were validated on independent sample sets.

[0116] S7. Clinical Applications;

[0117] Translate research findings into clinical applications, such as developing osteoporosis risk assessment tools based on bile acid levels.

[0118] It should be noted that in the process of predicting bone mass using bile acids in this invention, the relative abundance of porcine deoxycholic acid can be obtained by detecting the bile acid content in the target population. The relative abundance of porcine deoxycholic acid is then used in the Pearson correlation analysis method in step S3 to calculate the theoretical bone mineral density. This theoretical bone mineral density is then compared with the actual measured bone mineral density. If the actual measured bone mineral density is higher than the theoretically calculated bone mineral density, it is considered normal. If the actual measured bone mineral density is lower than the theoretically calculated bone mineral density, there is a risk of osteoporosis.

[0119] It should be noted that, through experiments, it was found that the porcine deoxycholic acid (POC) of the present invention... Hyodesoxycholic Acid HDCA The expression of this substance was lower in older mice than in younger mice, and both were strongly positively correlated with bone quality.

[0120] The present invention also provides a bile acid comprising porcine deoxycholic acid.

[0121] This invention also provides related products of porcine deoxycholic acid, including porcine deoxycholic acid and conventional excipients. The conventional excipients include: diluents such as lactose, microcrystalline cellulose, and starch; used to increase volume and improve compressibility; binders such as starch paste, cellulose derivatives, povidone, and gelatin; used to improve particle properties; disintegrants such as croscarmellose sodium and corn starch; used to promote tablet disintegration in the gastrointestinal tract; and lubricants such as magnesium stearate, micronized silica gel, and talc; used to reduce interparticle friction.

[0122] The process for preparing porcine deoxycholic acid and related products using bile acids includes the following steps:

[0123] S1. Preparation of HDCA;

[0124] S11. Raw material acquisition;

[0125] HDCA can be extracted from pig bile or produced through chemical or biosynthetic methods.

[0126] S12. Extraction and purification;

[0127] HDCA was extracted from the raw material using organic solvent extraction and chromatography, and then purified.

[0128] In step S12 above, the process of extracting hyodeoxycholic acid (HDCA) from porcine bile includes the following steps:

[0129] S121. Raw material preparation: Take fresh pig bile, or use the mother liquor from the bilirubin extraction process after filtering out bile calcium salts.

[0130] S122. Saponification: Mix bile with sodium hydroxide, add water, and saponify at 95-100℃ for 16-24 hours. Saponification is the process of converting bile acids into their sodium salts to facilitate subsequent extraction.

[0131] S123. Acidification: After saponification, cool to room temperature and let stand. Siphon off the supernatant, add water to the lower paste and stir to dissolve. Then acidify with a strong acid (such as hydrochloric acid or sulfuric acid) until Congo red test paper turns blue. This step converts the saponified bile acids into free bile acids.

[0132] S124. Extraction: Add ethyl acetate and stir to extract for 20-50 minutes. After standing and separating the layers, discard the aqueous phase and proceed with the subsequent processing of the organic phase (containing bile acids).

[0133] S125. Washing: Wash the ethyl acetate phase with water until the pH of the aqueous phase is 6-7.

[0134] S126. Drying and purification: The organic phase is dried with anhydrous sodium sulfate, and then crude porcine deoxycholic acid is obtained through steps such as filtration and concentration.

[0135] S127. Crystallization and Refining: The crude product is crystallized and refined to obtain high-purity porcine deoxycholic acid.

[0136] S128. Quality Control: The purity of bile acids is tested using analytical methods such as HPLC to ensure product quality.

[0137] Through the above processing, high-purity porcine deoxycholic acid can be extracted, and its purity is greater than 98% as determined by HPLC.

[0138] S2, Formulation Development;

[0139] Purified HDCA can be combined with conventional excipients to develop pharmaceutical formulations, such as tablets and capsules.

[0140] S3, Quality Control;

[0141] Quality control of the formulation is carried out, including determining the HDCA content; the animal experiment is 50 mg / kg / day.

[0142] The prepared porcine deoxycholic acid products were used in preclinical studies: pharmacodynamic and toxicological studies were conducted in animal models to assess the safety and efficacy of the drug.

[0143] Then, clinical trials are conducted to verify the safety and efficacy of the drug in humans.

[0144] After completing clinical trials, a new drug application is submitted to the drug regulatory agency. Once approved, large-scale production is carried out to obtain porcine deoxycholic acid products.

[0145] In step 1 above, HDCA can also be prepared by the following method, which specifically includes the following process:

[0146] S11. Raw material preparation;

[0147] Fresh pig bile is selected as the raw material.

[0148] S12, saponification;

[0149] Pig bile is mixed with a strong alkaline aqueous solution and saponified at 95-100℃ for 16-24 hours. The purpose of bile acid saponification is to extract and purify bile acids. This helps to improve the purity of bile acids, laying the foundation for subsequent purification and application.

[0150] S13, acidification;

[0151] After saponification, cool to room temperature and let stand. Then siphon out the supernatant. Add water to the lower paste and stir to dissolve. Add strong acid to acidify until Congo red test paper turns blue.

[0152] In step S3 above, the purpose of acidification after bile acid saponification is to convert the bile salts formed in the saponification reaction into free bile acids. Free bile acids are insoluble in water, and their precipitation can be achieved by adjusting the pH during the acidification process, thereby realizing the extraction and purification of bile acids.

[0153] S14, Extraction;

[0154] After acidification, ethyl acetate was added and the mixture was stirred and extracted for 20-50 minutes. After standing and separating into layers, the aqueous phase was discarded. The ethyl acetate phase was washed with water until the pH of the aqueous phase was 6-7, thus obtaining the organic phase.

[0155] S15, Drying;

[0156] The obtained organic phase was dried, and then concentrated and dried to obtain crude porcine deoxycholic acid.

[0157] S16, Refined;

[0158] The crude product is further purified by column chromatography, crystallization, or HPLC to improve product purity.

[0159] S17. Quality Inspection.

[0160] The obtained porcine deoxycholic acid was subjected to quality testing, including the detection of its purity and possible impurities.

[0161] The porcine deoxycholic acid of this invention is an organic compound with the molecular formula C. 24 H 40 O4 is a cholanic acid extracted from pig bile. It is a white or slightly yellowish powder with a bitter taste and a slightly fishy odor. It is slightly soluble in alcohol, sparingly soluble in acetone, very sparingly soluble in ether and chloroform, and almost insoluble in water.

[0162] Compared with existing technologies, the abundance of porcine deoxycholic acid can serve as a predictive indicator and can also be supplemented to combat osteoporosis. Therefore, this invention can use the abundance of porcine deoxycholic acid in bile acids to predict osteoporosis.

[0163] Example 1

[0164] This embodiment is based on the above-mentioned application of bile acids in predicting bone mass and preparing anti-osteoporosis drugs, and is used to explore the relationship between porcine deoxycholic acid and osteoporosis.

[0165] Grouping: The control group consisted of young mice (2 months old, n=20), and the experimental group consisted of older mice (18 months old, n=20). In this embodiment, the levels of porcine deoxycholic acid (HDCA) in serum samples from young and older mice were first detected by bile acid-targeted metabolomics sequencing. The average level in older mice was approximately 8.77 ng / ml, and the average level in young mice was approximately 20.77 ng / ml. Figure 1 As shown, the HDCA content in young mice was higher than that in older mice, and the difference was statistically significant.

[0166] We then used micro-CT to measure the bone microstructure of young and aged mice. The mean bone volume fraction of young mice was 0.3863, and the mean bone volume fraction of aged mice was 0.2596. The difference was statistically significant. Figure 2 Finally, we performed a correlation analysis between HDCA abundance and the bone volume fraction of the fifth lumbar vertebra. The results showed a strong positive correlation between HDCA abundance and the bone volume fraction of the fifth lumbar vertebra. Figure 3 The process of conducting correlation analysis includes the following steps:

[0167] S1. Data Preparation:

[0168] This invention provides the relative abundance and bone mineral density results of porcine deoxycholic acid obtained through mouse gut microbiota sequencing.

[0169] S2. Select Pearson correlation coefficient as the correlation analysis method and calculate the correlation coefficient.

[0170] S3. The obtained result R2 is 0.626, and the direction is positive correlation.

[0171] S4. The statistically significant difference in the results is p < 0.0001.

[0172] S5. Data Visualization: Use scatter plots to show the relationships between variables, helping to intuitively understand correlations (e.g., ...). Figure 3 (As shown).

[0173] Then, aged mice were treated with HDCA by gavage, and the groups were as follows: aged mice (18 months old) were randomly divided into two groups. One group was administered PBS by gavage as the control group (n=20), and the other group was administered HDCA (50 mg / kg / day) by gavage for 4 weeks (28 days) as the experimental group (n=20). After administration, dynamic bone mineral density testing showed ( Figure 4a In the first week, bone mineral density decreased in both groups (self-comparison, change from baseline). In the fourth week, the average decrease in bone mineral density in the HDCA-treated group was 0.4397%, while the average decrease in the control group was 2.59%, with a statistically significant difference (P=0.07). This indicates that HDCA has an anti-bone loss effect. Subsequently, micro-CT scans were performed on samples from the fifth lumbar vertebra (L5) of mice. Figure 4b The bone microstructure of the mice was assessed, and the results showed that the bone volume fraction of the aged mice treated with HDCA was higher than that of the aged mice treated with PBS (control group), and the difference was statistically significant. These results indicate that HDCA can improve bone mineral density (as shown in Figure 4).

[0174] Meanwhile, using TGR5 knockout mice, we demonstrated that the therapeutic effect of HDCA depends on the presence of TGR5 (TGR5 is a bile acid receptor, and bile acids need to bind to bile acid receptors to function). The experimental design is as follows;

[0175] Grouping: Wild-type mice (2-month-old C57 / BL6 mice, n=20) and TGR5 gene knockout mice (2-month-old, n=20), both groups were administered HDCA (50 mg / kg / day) by gavage for 4 weeks. Dynamic bone mineral density results ( Figure 5aThe results showed that at week four, bone mineral density increased in wild-type mice treated with HDCA (self-control, change from baseline), while it decreased in TGR5 knockout mice. This indicates that the anti-bone loss effect of HDCA depends on TGR5. Micro-CT scans were performed on samples from the fifth lumbar vertebra (L5). Figure 5b The bone microstructure of mice was assessed. The results showed that at week four, the bone volume fraction of wild-type mice treated with HDCA was significantly higher than that of TGR5 knockout mice. These results indicate that the effect of HDCA in improving bone microstructure in mice is dependent on TGR5.

[0176] In summary, HDCA levels in C57 / BL6 mice show a decreasing trend with age; therefore, HDCA is an indicator of aging. Furthermore, HDCA is strongly positively correlated with bone mass; higher serum HDCA levels are associated with denser lumbar vertebral microstructure; therefore, HDCA is a therapeutic target for osteoporosis. Further, our gavage experiment in aged mice demonstrated that HDCA can resist bone loss and increase bone mineral density.

[0177] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any changes or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention.

Claims

1. The use of a bile acid in the preparation of a drug for treating osteoporosis in elderly individuals, characterized in that, The bile acid is hyodesoxycholic acid (HDCA), and the drug is an oral preparation that inhibits bone loss and improves bone microstructure through oral administration.

2. The application according to claim 1, wherein, The drug is in the form of tablets or capsules.

3. The application according to claim 1, wherein, The drug contains conventional pharmaceutical excipients, which are selected from one or more of lactose, microcrystalline cellulose, starch, povidone, gelatin, magnesium stearate, and talc.

4. The application according to claim 1, wherein, The dosage of the porcine deoxycholic acid is 50 mg / kg / day.

5. The application according to claim 1, wherein, The porcine deoxycholic acid exerts its function in dependence on the bile acid receptor TGR5.

6. The application according to claim 1, wherein, The method for preparing the drug includes the following steps: (1) Extract and purify porcine deoxycholic acid from porcine bile, or obtain porcine deoxycholic acid by chemical or biological synthesis methods; (2) The purified porcine deoxycholic acid is mixed with conventional pharmaceutical excipients, wherein the excipients are selected from one or more of lactose, microcrystalline cellulose, starch, povidone, gelatin, magnesium stearate, and talc. (3) Prepare the resulting mixture into an oral pharmaceutical preparation, including tablets or capsules.

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