Application of polypeptide hormone ELABLA-21 in preparation of weight-losing product and medicine
By using polypeptide hormone ELABELA-21 to combine with auxiliary materials, the problem of poor treatment effect of existing weight loss drugs and large side effects of toxicity is solved, and a significant weight loss effect and smaller side effects are achieved.
Patent Information
- Application Number
- CN202411551786.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-01
- Publication Date
- 2025-01-28
Smart Images
Figure CN119367511A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of medicine and disease treatment, and specifically relates to the application of polypeptide hormone ELABELA-21 in the preparation of weight loss products and medicine. Background Art
[0002] As society continues to develop and progress, people's living standards are improving, and obesity has become more and more common. Complications of obesity, such as hypertension, type II diabetes, cardiovascular and cerebrovascular diseases, and some tumors, have affected human health and life satisfaction. How to prevent and treat obesity has become increasingly important.
[0003] At present, the two main strategies for the treatment of obesity in clinical practice are surgery and drug therapy. However, due to the high risk of surgical treatment, drug therapy is often chosen. There are various weight loss drugs on the weight loss market, mainly Western medicine, including drugs that increase energy metabolism such as thyroxine tablets, appetite suppressants such as drugs that inhibit intestinal digestion and absorption such as orlistat, and other drugs for the treatment of obesity such as insulin sensitizers. These expensive Western medicines have certain therapeutic effects on some people, but for most people, fat loss is just an illusion. After stopping use, the weight will rise again. At the same time, the toxic side effects are very large, which can cause gastrointestinal dysfunction, respiratory tract infection, headache, anxiety, cardiovascular disease, organ failure and even death.
[0004] ELABELA, a small molecule peptide hormone, has a homologous sequence with Apelin. As a ligand of the G protein-coupled receptor APJ, it plays an extremely important role in various pathological and physiological processes in the embryonic and adult stages. A large number of studies have shown that ELABELA has the effects of promoting angiogenesis, relaxation and cardiac development, and has a significant protective effect on cardiovascular diseases such as heart failure, hypertension, atherosclerosis and vascular remodeling. At present, it is necessary to develop new uses for ELABELA in weight loss. Among them, the full English name of APJ is ApelinPeptide Jejunum. Summary of the invention
[0005] The purpose of the present invention is to provide a new use of ELABELA in weight loss products. The present invention provides the use of polypeptide hormone ELABELA-21 in the preparation of weight loss products and medicines.
[0006] To achieve the above purpose, the present invention adopts the following technical solution.
[0007] The present invention provides a new application of polypeptide hormone ELABELA-21 in the preparation of weight loss products.
[0008] The amino acid sequence of the polypeptide hormone ELABELA-21 is shown in SEQ ID NO. 1: LRKHNCLQRRCMPLHSRVPFP. The polypeptide hormone ELABELA-21 is referred to as ELA-21.
[0009] The polypeptide hormone ELA-21 can reduce body weight.
[0010] The polypeptide hormone ELA-21 can reduce the fat content of the body.
[0011] The polypeptide hormone ELA-21 can inhibit the hypertrophy of white fat and brown fat cells.
[0012] The polypeptide hormone ELA-21 can improve glucose tolerance.
[0013] The polypeptide hormone ELA-21 can improve insulin resistance.
[0014] Preferably, the weight loss product comprises a weight loss food or a weight loss drug.
[0015] Preferably, the weight loss food contains the polypeptide hormone ELA-21 as an active ingredient and is supplemented with excipients acceptable in food science.
[0016] Preferably, the food science acceptable excipients include any one or more of starch, sucrose and cyclodextrin. The above excipients combine with ELA-21 to form a stable inclusion complex, increase the stability of ELA-21, protect ELA-21 molecules from environmental factors, and improve the bioavailability of ELA-21.
[0017] Preferably, the weight loss drug uses the polypeptide hormone ELA-21 as an active ingredient and is supplemented with pharmaceutically acceptable excipients.
[0018] Preferably, the pharmaceutically acceptable excipients include any one or more of starch, sucrose, trehalose, mannitol and sorbitol. The above excipients are carbohydrate excipients, which act as protein stabilizers and are combined with ELA-21 to significantly stabilize the secondary structure of the ELA-21 polypeptide, the extension and aggregation of the polypeptide chain during the freeze-drying process and the storage period, thereby enhancing the stability and bioavailability of ELA-21.
[0019] Preferably, the dosage form of the weight loss drug includes any one of tablets, capsules, granules, solutions and injections.
[0020] Preferably, the weight loss drug is a tablet. The auxiliary materials of the tablet include starch and sucrose, ELA-21, starch and sucrose are mixed to obtain a mixed powder; the mixed powder is mixed with starch slurry to obtain a soft material, which is then granulated, dried and granulated in sequence, and the obtained granules are mixed with talcum powder and then tableted to obtain the tablet.
[0021] Preferably, the weight loss drug is a capsule. The auxiliary material of the capsule includes sucrose, and the mixed powder of ELA-21 and sucrose is sieved and filled into a soft capsule material made of gelatin to obtain the capsule.
[0022] Preferably, the weight loss drug is a granule. The auxiliary materials of the granule include sucrose and dextrin, ELA-21, sucrose and dextrin are mixed to obtain a mixed powder; the mixed powder is mixed with an ethanol aqueous solution to obtain a soft material, which is then granulated, dried and granulated in sequence to obtain the granule.
[0023] Preferably, the weight loss drug is a solution.
[0024] The auxiliary materials used in the solution include solvents.
[0025] The solvent is any one or more of DMSO, water, ethanol, glycerol, and physiological saline.
[0026] The present invention also provides a medicine for weight loss, which is obtained by compounding the polypeptide hormone ELA-21 and a solvent.
[0027] Preferably, the solvent is any one or more of DMSO, water, ethanol, glycerol, and physiological saline.
[0028] Preferably, the concentration of the polypeptide hormone ELA-21 in the drug is 0.05 mg / mL to 0.1 mg / mL.
[0029] Compared with the prior art, the present invention has the following beneficial effects:
[0030] The present invention provides the use of polypeptide hormone ELA-21 in the preparation of weight loss products and medicine, and provides a new method for treating obesity. The present invention proves the positive therapeutic effect of polypeptide hormone ELA-21 in obesity through in vivo experiments (obese mice induced by high-fat diet). Specifically, polypeptide hormone ELA-21 significantly reduces the weight of obese mice, reduces fat accumulation, improves glucose tolerance and insulin resistance of mice, inhibits fat cell hypertrophy, and provides candidate drugs for the treatment of obesity and related symptoms. Moreover, it is an endogenous hormone synthesized by the body itself, and has less toxic side effects than existing weight loss drugs, and has made significant progress compared with the prior art. BRIEF DESCRIPTION OF THE DRAWINGS
[0031] Figure 1 This is the experimental process of the animal experiment in the embodiment of the present invention.
[0032] Figure 2 The results of the mouse GTT and ITT experiments in the present invention, wherein: Figure 2 Figure a in the figure is the fasting blood glucose of mice in the present invention; Figure 2 Figure b shows the blood glucose level-time curve of mice in each group in the GTT experiment; Figure 2 Figure c in the figure is the AUC of the blood glucose level-time curve of each group of mice in the GTT experiment of the present invention; Figure 2 Figure d in the figure is the blood glucose level-time curve of mice in each group of ITT experiment; Figure 2 Figure e in the figure is the AUC of the blood glucose level-time curve of each group of mice in the ITT experiment. GTT refers to glucose tolerance test, ITT refers to insulin tolerance test, and AUC refers to the area under the curve.
[0033] Figure 3 is the weight result of mice in the present invention, wherein, Figure 3 Figure a shows the weight changes of mice; Figure 3 Figure b shows the average daily food intake of mice; Figure 3 Figure c shows abdominal fat in mice.
[0034] Figure 4 HE staining results of mouse adipose tissue in the present invention: Figure 4 Figure a in the middle shows the HE staining result of white fat. Figure 4 Figure b shows the HE staining result of brown fat. DETAILED DESCRIPTION
[0035] The present invention is described in detail below in conjunction with the accompanying drawings and specific examples, but should not be construed as limiting the present invention. Unless otherwise specified, the technical means used in the following examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following examples, unless otherwise specified, can be obtained from commercial sources.
[0036] The mice used in the embodiments of the present invention are all of the C57BL / 6J strain and are raised in the Experimental Animal Center of Jiangxi University of Traditional Chinese Medicine in a specific pathogen-free animal room. Experimental Animal Use License No.: SCXK(Gan)2023-0001.
[0037] Among them, the English name of specific pathogens is specific pathogen free, SPF.
[0038] All mice used in the experiment were 8-week-old male mice, and all mouse experiments complied with the "Regulations on the Use of Experimental Animals" formulated by the National Ethics Committee and the Jiangxi University of Traditional Chinese Medicine. The mice used in the experiment are referred to as C57BL / 6J mice below.
[0039] Example 1
[0040] 1. Establishment, grouping and drug administration of obesity model
[0041] (1) Establishment of obesity model
[0042] After C57BL / 6J mice were fed a high-fat diet (45 kcal% fat, HFD) for 12 weeks, the weight of the mice increased by more than 30% compared with the control group mice, indicating that the modeling was successful, and the C57BL / 6J mouse model group mice were obtained.
[0043] (2) Grouping
[0044] Seventeen mice were randomly divided into three groups: control group, 5 mice; HFD group, 6 mice; HFD+ELA-21 group, 6 mice. The mice in the HFD+ELA-21 group were intraperitoneally injected with ELA-21 once a day starting from the 13th week, and samples were collected at the 16th week for subsequent experiments.
[0045] Among them, the control group is also called the Control group.
[0046] The experimental process of animal experiments is as follows Figure 1 shown.
[0047] The specific grouping and dosing methods are as follows:
[0048] Control group: The mice were fed with normal mouse diet and given an intraperitoneal injection of an equal volume of saline. Here, "equal volume" means that the volume of the injected saline is the same as the volume of the injected ELA-21 solution.
[0049] HFD group: fed with a high-fat diet, and given an intraperitoneal injection of an equal volume of saline. Here, "equal volume" means that the volume of the injected saline is the same as the volume of the injected ELA-21 solution.
[0050] HFD+ELA-21 group: The mice were fed with a high-fat diet and given an intraperitoneal injection of ELA-21 solution. The amount of ELA-21 injected into the mice was 1 mg / kg / day.
[0051] Among them, ELA-21 was synthesized by Nanjing GenScript Biotechnology Co., Ltd. with a purity of >98% and stored at -20°C. Before use, it was dissolved in physiological saline and sterilized with a 0.22μm filter to obtain an ELA-21 solution with a concentration of 0.06mg / mL. ELA-21 is the polypeptide hormone ELABELA-21. The amino acid sequence of the polypeptide hormone ELABELA-21 is shown in SEQ ID NO.1: LRKHNCLQRRCMPLHSRVPFP.
[0052] 2. Glucose tolerance and insulin tolerance test
[0053] After overnight fasting, the mice in each group were given glucose by gavage, and the blood glucose levels were tested after 0, 15, 30, 60, 90 and 120 minutes. The amount of glucose given was 1 g / kg.
[0054] After fasting mice were intraperitoneally injected with 0.75U / Kg insulin, blood glucose levels were measured at 0, 15, 30, 60, 90, and 120 minutes later. Figure 2 shown.
[0055] Among them, the method for detecting blood sugar level is: cut 2mm from the tip of the mouse tail, discard the first drop of blood, and use Sinocare GA-3 blood sugar meter and corresponding blood sugar test strips to detect the blood sugar level of the second drop of blood.
[0056] Among them, the fasting blood glucose of mice Figure 2 As shown in Figure a; The blood glucose level-time curves of mice in each group of GTT experiment are shown in Figure 2 As shown in Figure b; AUC of the blood glucose level-time curve of each group of mice in the GTT experiment Figure 2 As shown in Figure c; The blood glucose level-time curves of mice in each group of ITT experiment are shown in Figure 2 As shown in Figure d; the AUC of the blood glucose level-time curve of each group of mice in the ITT experiment is as follows Figure 2 As shown in Figure e. GTT refers to glucose tolerance test, ITT refers to insulin tolerance test, and AUC refers to area under the curve. There was no significant difference in the fasting blood glucose levels of mice in each group. Compared with the control group, the blood glucose levels and AUC values of mice in the HFD group at each time point were significantly increased after oral administration of glucose or intraperitoneal injection of insulin, and the blood glucose levels and AUC values at each time point were significantly reduced after intervention with ELA-21.
[0057] Depend on Figure 2 It was found that ELA-21 treatment significantly improved glucose tolerance and insulin resistance induced by a high-fat diet in mice.
[0058] 3. Sample collection and processing
[0059] The mice in each group obtained at the end of the experiment were fasted for 12 hours before sampling, but not water deprivation, and weighed, and their eyeballs were removed to collect blood. The abdominal skin of the mice was cut open with surgical scissors, and the gross images were taken with a camera.
[0060] The weight changes of mice were as follows Figure 3 As shown in Figure a, the average daily food intake of mice is Figure 3 As shown in Figure b, the abdominal fat of mice is Figure 3 As shown in Figure c. Compared with the control group, the weight of mice fed a high-fat diet increased significantly, and the abdominal fat increased significantly; after intervention with ELA-21, the weight of mice was significantly reduced, and the body fat was reduced, but it had no effect on the diet of mice.
[0061] Depend on Figure 3 It can be seen that ELA-21 treatment significantly reduced the body weight and body fat content of mice, indicating that ELA-21 has a significant weight loss effect.
[0062] 1.5 cm×1.0 cm×0.2 cm adipose tissue of mice in the control group, HFD group and HFD+ELA-21 group was fixed in 4% by mass paraformaldehyde, and the remaining adipose tissue was quick-frozen in liquid nitrogen for 30 min and then stored at -80°C for later use.
[0063] The mouse blood collected from each group was placed in a 1.5 mL centrifuge tube, allowed to stand at room temperature for 1 hour, centrifuged at 4°C and 3500 r / min for 10 minutes, the supernatant was collected to obtain serum samples, and stored at -80°C for later use.
[0064] 4. HE staining of adipose tissue
[0065] After the adipose tissue in step 3 is fixed in a 4% by volume paraformaldehyde solution at 4°C for 24 hours, the adipose tissue is taken out and rinsed with running water, and then embedded in paraffin after automatic dehydration and wax immersion. The slices are sliced with a paraffin slicer with a thickness of 4 μm, baked in an oven at 65°C for 2 hours, and stored at room temperature for later use to obtain paraffin slices.
[0066] Paraffin sections were dewaxed to water by an automatic stainer, and then stained with hematoxylin for 5 minutes, washed with distilled water, differentiated with differentiation solution, washed with distilled water, and blued with blueing solution until the cytoplasm was dark blue and the nucleus was not stained, rinsed with running water, and the sections were sequentially dehydrated with 85% ethanol and 95% ethanol for 5 minutes each, and stained with eosin solution for 5 minutes. The sections were then sequentially placed in 100% ethanol I for 3 minutes, 100% ethanol II for 3 minutes, 100% ethanol III for 3 minutes, xylene I for 3 minutes, and xylene II for 3 minutes; the sections were sealed with neutral gum and photographed under a microscope. The results are shown in the figure. Figure 4 shown.
[0067] Among them, the HE staining results of white fat are as follows Figure 4 As shown in Figure a, the HE staining results of brown fat are as follows Figure 4 As shown in Figure b, compared with the control mice, the size of white fat and brown fat cells in mice fed a high-fat diet was significantly increased, while ELA-21 intervention significantly reduced the size of fat cells.
[0068] Depend on Figure 4 It can be seen that ELA-21 treatment significantly inhibited the hypertrophy of white fat and brown fat cells in mice.
[0069] From the above experimental results, it can be seen that ELA-21 can significantly reduce the body weight of obese mice, reduce fat accumulation in the body, improve glucose tolerance and insulin resistance of mice, inhibit fat cell hypertrophy, and has a positive therapeutic effect on obesity.
[0070] It should be noted that when the present invention involves a numerical range, it should be understood that the two endpoints of each numerical range and any numerical value between the two endpoints can be selected. In order to avoid redundancy, the present invention describes a preferred embodiment.
[0071] Although preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they understand the basic inventive concepts, and all such changes and modifications fall within the scope of the present invention.
Claims
1. The use of polypeptide hormone ELABELA-21 in the preparation of weight loss products, characterized in that: The amino acid sequence of the polypeptide hormone ELABELA-21 is shown in SEQ ID NO.
1.
2. The use according to claim 1, characterized in that: The slimming products include slimming foods or slimming drugs.
3. The use according to claim 2, characterized in that: The weight-loss food uses the polypeptide hormone ELABELA-21 as an active ingredient and is supplemented with auxiliary materials acceptable in food science.
4. The use according to claim 3, characterized in that: The excipients acceptable in food science include any one or more of starch, sucrose and cyclodextrin.
5. The use according to claim 2, characterized in that: The weight loss drug uses the polypeptide hormone ELABELA-21 as an active ingredient and is supplemented with pharmaceutically acceptable excipients.
6. The use according to claim 5, characterized in that: Pharmaceutically acceptable excipients include any one or more of starch, sucrose, trehalose, mannitol and sorbitol.
7. The use according to claim 5, characterized in that: The dosage form of the weight loss drug includes any one of tablets, capsules, granules, solutions and injections.
8. The use according to claim 5, characterized in that: The weight loss drug is a solution; The auxiliary materials supplemented with the solution include solvents; The solvent is any one or more of DMSO, water, ethanol, glycerol, and physiological saline.
9. A drug for weight loss, characterized in that: The drug is prepared by compounding the polypeptide hormone ELABELA-21 described in claim 1 with a solvent; The solvent is any one or more of DMSO, water, ethanol, glycerol, and physiological saline.
10. The drug according to claim 9, characterized in that The concentration of the polypeptide hormone ELABELA-21 in the drug is 0.05 mg / mL to 0.1 mg / mL.