Application of bile acid in preparation of medicine for treating tumor cachexia
By using bile acids, especially deoxycholic acid, therapeutic drugs were prepared to inhibit lipid degradation-related proteins, solving the problem of fat tissue consumption in tumor cachexia and achieving significant weight recovery and improvement of fat tissue condition.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- NANJING UNIV
- Filing Date
- 2026-02-28
- Publication Date
- 2026-05-08
AI Technical Summary
Existing treatments for cancer cachexia are unable to effectively reverse the consumption of adipose tissue, especially fat breakdown, leading to depletion of energy reserves and exacerbation of systemic inflammation. Furthermore, the mechanisms of existing therapies are not yet fully understood, limiting the development of targeted treatment strategies.
Using bile acids, particularly deoxycholic acid from secondary bile acids, as active ingredients, therapeutic drugs are prepared to inhibit the levels of lipid degradation-related proteins and improve weight loss and adipose tissue depletion symptoms in patients with cancer cachexia.
Deoxycholic acid significantly restored body weight in mice with tumor cachexia, increased adipose tissue weight and cross-sectional area, reduced serum triglyceride and free fatty acid levels, inhibited adipocyte degradation, and improved abnormal adipose tissue breakdown.
Smart Images
Figure CN121987643A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to tumor cachexia, and more particularly to the use of a bile acid in the preparation of a drug for treating tumor cachexia. Background Technology
[0002] Cancer cachexia is a multifactorial metabolic disorder syndrome caused by cancer, mainly characterized by progressive, involuntary weight loss, especially the continuous consumption of skeletal muscle and adipose tissue, often accompanied by loss of appetite, fatigue, and systemic inflammatory response. Abnormal lipolysis is one of the core features of cachexia; even with sufficient calorie intake, patients experience a rapid reduction in systemic fat, which is completely different from the fat consumption pattern under normal starvation. Lipolysis not only leads to depletion of energy reserves but also releases large amounts of free fatty acids, further exacerbating insulin resistance, hepatic steatosis, and systemic low-grade chronic inflammation, forming a vicious cycle of metabolic imbalance and tissue consumption. Furthermore, adipose tissue is not only an energy storage organ but also has endocrine functions; its abnormal breakdown can also affect the secretion of adiponectin, leptin, and other adipose-derived hormones, indirectly participating in the regulation of appetite and energy metabolism.
[0003] Currently, treatment options for cancer cachexia are limited and their effectiveness varies. The main strategies include nutritional support, drug therapy, and anti-tumor therapy. While nutritional support can improve weight in some patients, its use alone often fails to reverse the progressive loss of muscle and fat. Regarding drugs, progesterone derivatives can stimulate appetite but may cause side effects such as blood clots; glucocorticoids improve appetite and energy in the short term, but long-term use can accelerate muscle loss. Overall, existing therapies primarily focus on symptom relief and cannot effectively reverse fat loss. The fundamental reason is that cachexia involves multiple pathway interactions, and the mechanisms are not yet fully understood. In particular, the specific signaling regulatory network for fat degradation still needs further exploration, limiting the development of targeted treatment strategies. Summary of the Invention
[0004] Purpose of the invention: The purpose of this invention is to provide a bile acid in the preparation of a drug for the treatment of tumor cachexia, especially a drug for the treatment of symptoms such as weight loss and depletion of adipose tissue in patients with tumor cachexia.
[0005] Technical solution: The application of bile acids described in this invention in the preparation of drugs for the treatment of tumor cachexia.
[0006] Preferably, the bile acid is a secondary bile acid; more preferably, the secondary bile acid is deoxycholic acid.
[0007] Preferably, the application is in the preparation of a drug for improving the weight of patients with tumor cachexia.
[0008] Preferably, the application is in the preparation of a drug for treating adipose tissue depletion symptoms of tumor cachexia; more preferably, the adipose tissue depletion symptoms include fat decomposition and browning of white adipose tissue.
[0009] Preferably, the drug contains bile acids or their pharmaceutically acceptable salts, solvates, hydrates, or metabolites as active ingredients.
[0010] Preferably, the drug further contains pharmaceutically acceptable excipients; more preferably, the pharmaceutically acceptable excipients include any one or more of excipients, diluents, lubricants, flow aids, wetting agents, emulsifiers, pH buffers, solubilizers, cosolvents, or solvents.
[0011] Preferably, the dosage form of the drug includes tablets, capsules, granules, powders, chewable tablets, effervescent tablets, sustained-release tablets, microcapsules, injections, infusions, suspensions, patches, suppositories, transdermal patches, microemulsions, liposomes, and nanoparticles.
[0012] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: The present invention proposes and verifies for the first time that bile acids, especially deoxycholic acid in secondary bile acids, can effectively restore the body weight of mice with tumor cachexia model, and increase the weight and cross-sectional area of adipose tissue and increase the size of lipid droplets in adipocytes by inhibiting the level of lipid degradation-related proteins, thus providing a highly promising therapeutic drug for the symptoms of weight loss and adipose tissue depletion caused by tumor cachexia. Attached Figure Description
[0013] Figure 1 This is a statistical graph showing the rate of weight change in C26 tumor cachexia-induced lipid degradation mouse model after different treatments, where ** indicates p <0.01; Figure 2 This is a statistical graph showing the final tumor-free body weight of mice in the C26 tumor cachexia-induced lipid degradation model after different treatments, where **** indicates... p <0.0001; Figure 3 Representative images of epididymal adipose tissue in mice with C26 tumor cachexia-induced adipose degradation model after different treatments; Figure 4 This is a statistical graph showing the epididymal adipose tissue weight in C26 tumor cachexia-induced adipose degradation model mice after different treatments, where **** indicates... p <0.0001; Figure 5 This is an analysis of adipose tissue in C26 tumor cachexia-induced adipose degradation model mice after different treatments. A is a micrograph of adipocytes, B is a statistical graph of adipose tissue cross-sectional area, and **** indicates... p<0.0001; Figure 6 This is a statistical graph showing the serum triglyceride levels in C26 tumor cachexia-induced lipid degradation model mice after different treatments, where **** indicates... p <0.0001; Figure 7 This is a statistical graph showing the serum free fatty acid content in C26 tumor cachexia-induced lipid degradation model mice after different treatments, where **** indicates... p <0.0001; Figure 8 This figure shows the results of measuring the levels of lipid degradation-related proteins in C26 tumor cachexia-induced lipid degradation model mice after different treatments. Figure A shows the Western blot results, and figure B shows the relative grayscale value statistics. * indicates... p <0.05, ** indicates p <0.01; Figure 9 This figure shows the results of lipid degradation-related protein levels measured after different treatments in a C26 tumor cachexia-induced lipid degradation cell model. Figure A shows the Western blot results, and figure B shows the relative grayscale statistical results. ** indicates... p <0.01, *** indicates p <0.001, **** indicates p <0.0001; Figure 10 Representative Oil Red O staining images of a C26 tumor cachexia-induced lipid degradation cell model after different treatments. Detailed Implementation
[0014] The technical solution of the present invention will be further described below.
[0015] Example 1: Validation of the effect of deoxycholic acid on C26 tumor cachexia-induced lipid degradation model mice Male BALB / C mice aged 6-8 weeks were purchased from Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd. All mice were randomly divided into four groups: control group, normal drug administration group (control group + deoxycholic acid), cachexia group (C26), and deoxycholic acid treatment group (C26 + deoxycholic acid), with five mice in each group.
[0016] Mice in the C26 group and the C26+deoxycholic acid group were inoculated with 100 μL of a solution containing 5×10⁻⁶ mcg on their right side. 5 A suspension of C26 cells (provided by Nanjing University) was used. Mice in the control group and the control + deoxycholic acid group were injected with only 100 μL of PBS.
[0017] All mice were treated starting on day 7. Mice in the deoxycholic acid treatment group and the normal treatment group were administered 30 mg / kg of deoxycholic acid (purchased from MedChemExpress LLC., catalog number HY-N0593) by gavage in DMSO-corn oil (v:v=1:9). Mice in the cachexia group and the control group were given an equal volume of DMSO-corn oil (v:v=1:9). The drugs were administered once daily starting on day 7 when tumors began to grow, for a total of 7 administrations. The end of the administration period was day 14. The corn oil was purchased from MedChemExpress LLC., catalog number HY-Y1888.
[0018] 1. Weight monitoring The mice in each group were weighed and their weights recorded every two days. The rate of change in body weight (W) was calculated using the following formula. c ): W c =(WW n ) / W n ×100%, where W represents the weight on the day of measurement, W n This indicates the initial weight before modeling.
[0019] The rate of change in body weight of mice in each group is as follows: Figure 1 As shown.
[0020] At the end of the experiment, the mice from which tumors were removed were weighed, and the results were as follows: Figure 2 As shown in Table 1.
[0021] Table 1. Terminal tumor-free body weight of mice
[0022] The above results indicate that, in the C26 tumor cachexia-induced fat degradation model mice, the weight loss in the deoxycholic acid treatment group was significantly improved compared to the cachexia group mice.
[0023] 2. Evaluation of fat status in mice At the end of the experiment, images of the white adipose tissue in the mouse epididymis were acquired, and the results are as follows. Figure 3 As shown.
[0024] It was then completely removed and weighed using an analytical balance, and the results were recorded as follows: Figure 4 As shown in Table 2.
[0025] Table 2. Terminal adipose tissue weight (g) in mice
[0026] The above results indicate that in the C26 tumor cachexia-induced fat degradation model mice, the fat weight of the deoxycholic acid treatment group mice was significantly increased compared with the cachexia group mice.
[0027] White adipose tissue collected from mouse epididymis was fixed by immersion in 4% paraformaldehyde solution, followed by gradient dehydration, paraffin embedding, sectioning, dewaxing, and staining with hematoxylin and eosin (H&E). After mounting with neutral resin, the tissue was observed and images were acquired under a microscope. Ten adipocytes were randomly selected from each tissue section, and the cross-sectional area of the adipose tissue was statistically analyzed using ImageJ software. The results are as follows: Figure 5 As shown in Table 3.
[0028] Table 3 Cross-sectional area of mouse adipose tissue (μm) 2 )
[0029] These results indicate that deoxycholic acid treatment can significantly improve lipid degradation in C26 tumor cachexia-induced lipid degradation mouse model mice.
[0030] 3. Detection of serum triglyceride and fatty acid content in mice At the end of the experiment, venous blood was collected from mice, and after serum separation, the serum triglyceride (TG) and free fatty acid (FFA) levels were measured using the Solarbio Serum Triglyceride Assay Kit (catalog number BC0625) and the Solarbio Free Fatty Acid Assay Kit (catalog number BC0595) according to the kit instructions. Results are as follows: Figure 6 , Figure 7 As shown in Tables 4 and 5.
[0031] Table 4. Serum triglyceride levels (mM) in mice at the terminal stage
[0032] Table 5. Serum free fatty acid content (mM) in mice at the terminal stage
[0033] The above results indicate that in the C26 tumor cachexia-induced fat degradation model mice, deoxycholic acid treatment can significantly reduce fat loss in cachexia mice compared to the cachexia group mice.
[0034] 3. Determination of levels of lipid degradation-related proteins in mouse adipose tissue 30 mg of epididymal adipose tissue was collected, ground in liquid nitrogen using a mortar and pestle, and then lysed thoroughly with 300 μL of RIPA lysis buffer (product number P0013B). The lysis was performed on ice for 20 min, followed by centrifugation at 12000 rpm for 20 min to obtain the supernatant. Then, Beyotime SDS-PAGE protein loading buffer (product number P0015) was added according to the specified ratio, and the mixture was heated to boiling in a metal bath for 10 min. SDS-PAGE electrophoresis was then performed. After transfer and blocking, UCP1 primary antibody (purchased from Santa Cruz, product number A21979) diluted 1:1000 and ATGL primary antibody (purchased from Cell Signaling) diluted 1:1000 were added. Technology, catalog number 2138S), p-HSL primary antibody diluted 1:1000 (purchased from Wuhan Aiboteke Biotechnology Co., Ltd., catalog number AP1242) or β-actin primary antibody diluted 1:5000 (purchased from Hangzhou Fode Biotechnology Co., Ltd., catalog number FD0060) were incubated overnight at 4°C. After rinsing, the corresponding species secondary antibody diluted 1:10000 was incubated at room temperature for 2 h. Finally, the images were acquired and relative grayscale values were analyzed using Fode Biotechnology's ECL chemiluminescence kit (catalog number FD8020).
[0035] The results are as follows Figure 8 As shown, in C26 tumor cachexia-induced lipid degradation model mice, compared with cachexia group mice, deoxycholic acid treatment significantly reduced the expression of ATGL and UCP1 and the phosphorylation level of HSL, indicating that deoxycholic acid can effectively improve C26 tumor cachexia-induced lipid degradation.
[0036] Example 2: Validation of the effect of deoxycholic acid on the C26 tumor cachexia-induced lipid degradation cell model C26 cells were seeded into culture dishes. When the confluence reached about 70%, the original culture medium was discarded, and fresh RPMI 1640 complete culture medium was added and cultured for another 48 h. The culture supernatant was collected, centrifuged at 1000 ×g for 5 min to remove cells, and filtered through a 0.22 μm filter membrane to collect cell-free supernatant.
[0037] 3T3-L1 cells (provided by Nanjing University) were induced to differentiate into mature adipocytes using the MDI method and divided into four groups: control group, normal drug treatment group (control group + deoxycholic acid), cachexia group (C26), and deoxycholic acid treatment group (C26 + deoxycholic acid). For the cachexia group, the culture medium was replaced with cell-free supernatant containing 0.1% DMSO; for the deoxycholic acid treatment group, the culture medium was replaced with cell-free supernatant containing 0.1% DMSO and a final concentration of 10 μM deoxycholic acid; for the control group, 0.1% DMSO was added to the culture medium; and for the normal drug treatment group, 0.1% DMSO and a final concentration of 10 μM deoxycholic acid were added to the culture medium.
[0038] 1. Determination of levels of lipid degradation-related proteins After culturing for 24 h, cells from each group were collected, and 200 μL of Beyotime RIPA lysis buffer was added to completely lyse the tissues. After lysis on ice for 20 min, the cells were centrifuged at 12000 rpm for 20 min to obtain the supernatant. Subsequently, the expression levels of ATGL and the phosphorylation levels of HSL were measured according to the method described in Example 1.
[0039] The results are as follows Figure 9 As shown, in the C26 tumor cachexia-induced lipid degradation cell model, compared with the cachexia group, the ATGL expression and HSL phosphorylation levels of cells in the deoxycholic acid treatment group were significantly reduced, indicating that deoxycholic acid can improve C26 tumor cachexia-induced lipid degradation.
[0040] 2. Oil Red O staining After culturing for 24 h, the culture medium was discarded and the cells were washed with PBS buffer. After fixing with 4% paraformaldehyde solution at room temperature for 30 min, the cells were stained with Beyotime Oil Red O staining kit (C0157S) for 30 min. After washing away excess dye with PBS buffer, the cells were observed and images were acquired under a microscope.
[0041] The results are as follows Figure 10 As shown, in the C26 tumor cachexia-induced lipid degradation cell model, the lipid droplets in the deoxycholic acid treatment group were larger than those in the cachexia group, further indicating that deoxycholic acid can improve C26 tumor cachexia-induced lipid degradation.
Claims
1. The application of a bile acid in the preparation of a drug for treating tumor cachexia.
2. The application according to claim 1, characterized in that, The bile acids mentioned are secondary bile acids.
3. The application according to claim 2, characterized in that, The secondary bile acid is deoxycholic acid.
4. The application according to claim 1, characterized in that, The application is in the preparation of drugs to improve the weight of patients with tumor cachexia.
5. The application according to claim 1, characterized in that, The application is in the preparation of drugs for treating fat depletion symptoms of tumor cachexia.
6. The application according to claim 5, characterized in that, The symptoms of fat depletion include fat breakdown and browning of white adipose tissue.
7. The application according to claim 1, characterized in that, The drug contains bile acids or their pharmaceutically acceptable salts, solvates, hydrates, or metabolites as active ingredients.
8. The application according to claim 7, characterized in that, The drug also contains pharmaceutically acceptable excipients.
9. The application according to claim 8, characterized in that, The pharmaceutically acceptable excipients include any one or more of the following: excipients, diluents, lubricants, glidants, wetting agents, emulsifiers, pH buffers, solubilizers, cosolvents, or solvents.
10. The application according to claim 1, characterized in that, The dosage forms of the drugs include tablets, capsules, granules, powders, chewable tablets, effervescent tablets, sustained-release tablets, microcapsules, injections, infusions, suspensions, patches, suppositories, transdermal patches, microemulsions, liposomes, and nanoparticles.