Application of parthenolide and / or parthenolide derivative in preparation of medicine for preventing and / or treating diabetes mellitus and preparation of body weight intervention and / or blood sugar maintaining product

Abstract

The invention relates to the technical field of biological medicines, in particular to application of parthenolide and / or parthenolide derivatives in preparation of medicines for preventing and / or treating diabetes mellitus and in preparation of products for body weight intervention and / or blood sugar maintenance. The bitter compound parthenolide can be obtained from a natural product and is safe in component, the binding energy of parthenolide and GLP1R on the atomic level is-6.3 kcal / mol, parthenolide can serve as a GLP-1 receptor agonist and can promote GLP-1 secretion, the onset concentration of parthenolide is low (15-30 [mu] M), and it is indicated that parthenolide serves as a receptor to excite the secretion of intestinal peptide hormone, and the titer intensity of parthenolide is high. Therefore, parthenolide and / or parthenolide derivatives are / is a secretagogue which efficiently secretes GLP-1 and is low in side effect, are / is a promising direction for improving the endogenous GLP-1 level, provide a good scientific basis for developing products for effectively intervening body weight and maintaining the blood sugar health level in the future, and have great potential market value.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and in particular to the use of parthenolide and / or parthenolide derivatives in the preparation of drugs for preventing and / or treating diabetes, and in the preparation of products for weight intervention and / or blood glucose maintenance. Background Art

[0003] Glucagon-like peptide-1 (GLP-1) is an endocrine hormone mainly secreted by intestinal endocrine L cells, pancreatic α cells and nucleus tractus solitarii cells in the brainstem, which has the effects of lowering blood glucose and controlling body weight. It can lower blood glucose levels and control body weight by promoting insulin secretion, reducing glucagon content, inhibiting gastric emptying and reducing food intake to prevent T2 DM. At present, although there are many commercially available GLP-1 drugs that help to reduce body weight, GLP-1 drugs have some disadvantages, including: (1) Gastrointestinal side effects: GLP-1 drugs may cause gastrointestinal discomfort, such as side effects like nausea, vomiting and diarrhea, which may affect the quality of life of patients; (2) High cost: The prices of some GLP-1 drugs are relatively high, which may limit the use of some patients; (3) Long-term safety: Although the short-term safety of GLP-1 drugs is relatively good, some long-term safety issues, such as pancreatitis and thyroid tumors, still need further research and monitoring.

[0004] Therefore, it is crucial to provide a GLP-1 agonist with safe ingredients. Summary of the Invention

[0005] To solve the above problems, the purpose of the present invention is to provide the use of parthenolide and / or parthenolide derivatives in the preparation of drugs for preventing and / or treating diabetes, and in the preparation of products for weight intervention and / or blood glucose maintenance. The present invention discovers that parthenolide and / or parthenolide derivatives have a relatively high potency in causing the secretion of enteropeptide hormones as receptor agonists, and have great prospects in the research and development and application of blood glucose drugs and weight control.

[0006] The purpose of the present invention can be achieved by the following technical solutions:

[0007] The first purpose of the present invention is to provide the use of parthenolide and / or parthenolide derivatives in the preparation of drugs for preventing and / or treating diabetes, wherein the parthenolide and / or parthenolide derivatives are used as GLP-1 receptor agonists; the chemical structural formula of parthenolide is shown as follows:

[0008]

[0009] In one embodiment of the present invention, the drug is a drug that promotes the secretion of GLP-1 by cells, thereby promoting insulin secretion and reducing the content of glucagon.

[0010] In one embodiment of the present invention, the concentration of parthenolide and / or parthenolide derivative in the drug is 15 - 30 μM.

[0011] In one embodiment of the present invention, the concentration of parthenolide and / or parthenolide derivative in the drug is 15 μM.

[0012] The second object of the present invention is to provide a drug for preventing and / or treating diabetes, which contains parthenolide and / or parthenolide derivative.

[0013] The third object of the present invention is to provide the use of parthenolide and / or parthenolide derivative in the preparation of products for body weight intervention and / or blood glucose maintenance, and the parthenolide and / or parthenolide derivative is used as a GLP-1 receptor agonist.

[0014] In one embodiment of the present invention, the product is a product that inhibits gastric emptying and reduces food intake.

[0015] In one embodiment of the present invention, the concentration of parthenolide and / or parthenolide derivative in the product is 15 - 30 μM.

[0016] In one embodiment of the present invention, the concentration of parthenolide and / or parthenolide derivative in the product is 15 μM.

[0017] The fourth object of the present invention is to provide a product for body weight intervention and / or blood glucose maintenance, which contains parthenolide and / or parthenolide derivative;

[0018] The product is selected from one of food, cosmetics, health products or drugs.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] The bitter compound parthenolide can be obtained from natural products with safe ingredients. At the atomic level, the binding energy of parthenolide to GLP1R is -6.3 kcal / mol. Moreover, it can act as a GLP-1 receptor agonist, promoting GLP-1 secretion, and its effective concentration is relatively low (15 - 30 μM), indicating that parthenolide has a relatively high potency in inducing the secretion of intestinal peptide hormones as a receptor agonist. Therefore, parthenolide and / or parthenolide derivatives are secretagogues with high efficiency in secreting GLP-1 and low side effects, which will be a promising direction for increasing the endogenous GLP-1 level, providing a good scientific basis for the future development of products for effective weight intervention and maintaining a healthy blood glucose level, and having great potential market value. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 It is the molecular docking diagram of parthenolide and GLP1R in Example 1;

[0022] Figure 2 It is the test result diagram of the cell viability of different concentrations of parthenolide solution in Example 2;

[0023] Figure 3 It is the diagram of the influence of different concentrations of parthenolide on GLP-1 secretion in Example 2. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0024] The present invention will be described in detail below with reference to the drawings and specific embodiments.

[0025] In the following embodiments, Caco-2 cells (FH0029) were purchased from Shanghai Fuheng Biotechnology Co., Ltd., parthenolide (purity > 98%) was purchased from Shanghai Titan Technology Co., Ltd., DMEM high-glucose medium (4.5 g / L glucose) was purchased from Beijing Solarbio Science & Technology Co., Ltd., CCK8 cell proliferation detection kit was purchased from Shanghai Sheng'er Technology Co., Ltd., and human GLP-1 ELISA kit was purchased from Shenzhen Youpin Biotechnology Co., Ltd.; unless otherwise specified, all reagents used were commercially available reagents, and all detection means and methods used were conventional detection means and methods in the art.

[0026] Example 1

[0027] This example provides molecular docking to predict the binding ability of parthenolide to GLP1R.

[0028] (1) Based on the RCSB Protein Data Bank database (https: / / www.rcsb.org / ), retrieve the structures of the main active ingredients and download the PDB structure files. Open the crystal 3D structure file of the target protein - GLP1R (GLP-1 receptor) obtained from the PDB database using PyMOL 1.7.2.1 software. Remove water molecules, split the protein from its accompanying ligand, and save them separately in PDB format.

[0029] (2) Retrieve small molecule compounds based on the Pubchem database (https: / / pubchem.ncbi.nlm.nih.gov / ) and obtain their structure files in sdf format. Import the downloaded sdf format files into Chem3D 22.0.0.22 software and convert the small molecule compounds with 2D structures into 3D structures in mol2 format.

[0030] (3) Use AutoDock Tools 1.5.6 software to perform operations such as dehydrating, adding hydrogen, calculating charges, and combining non-polar hydrogens on the protein and small molecule compounds, and then save them as pdbqt format files. Use the virtual screening script to perform molecular docking on the small molecule compounds and the protein.

[0031] (4) After the docking is completed, the free binding energy of each small molecule compound binding to the protein can be obtained. The lower the binding energy, the better the docking effect. Select the conformation with the best affinity as the final docking conformation for visualization in the pyMOL software, further analyze the spatial structure and hydrogen bond interactions after binding, and draw the small molecule compound - protein docking image. Autodock-vina performs semi-flexible docking, and the obtained interaction energy results are as Figure 1 shown.

[0032] Performing molecular docking on parthenolide and GLP1R, its binding energy is -6.3 kcal / mol, indicating that parthenolide can dock well with GLP1R. Visualize it through the PyMOL software, as Figure 1 shown. Parthenolide forms a hydrogen bond with GLP1R at SERA 49. The results show that parthenolide binds well to the action target of GLP1R.

[0033] Example 2

[0034] This example provides the effects of different concentrations of parthenolide on the secretion of GLP-1 by intestinal endocrine cells Caco-2.

[0035] (1) Cell viability assay:

[0036] (101) Caco-2 cells were seeded in a 96-well plate with DMEM medium and incubated in a carbon dioxide cell incubator for 24 h.

[0037] (102) The supernatant in the 96-well plate was discarded, and parthenolide samples were serially diluted with DMEM (the concentrations of parthenolide were 0 μM, 3.75 μM, 7 μM, 15 μM, 30 μM, and 60 μM respectively). 100 μL of the diluted samples (the concentrations of parthenolide were 0 μM, 3.75 μM, 7 μM, 15 μM, 30 μM, and 60 μM respectively) were added to the 96-well plate and incubated in a carbon dioxide cell incubator for 24 h.

[0038] (103) The supernatant in the 96-well plate was discarded, and a CCK-8 assay kit was used to detect the viability of Caco-2 cells. (Caco-2 cells + diluted parthenolide sample + CCK-8 was recorded as the OD value of the experimental wells; Caco-2 cells + CCK-8 was recorded as the OD value of the blank wells; Caco-2 cells + DMEM medium + CCK-8 was recorded as the OD value of the control wells)

[0039] (104) Calculate the effect of the parthenolide solution on the viability of Caco-2 cells according to the following formula:

[0040] Cell survival rate = (OD value of experimental wells - OD value of blank wells) / (OD value of control wells - OD value of blank wells) × 100%;

[0041] The results were as Figure 2 shown. The viability of Caco-2 cells was determined with parthenolide at different concentrations (the concentrations of parthenolide were 0 μM, 3.75 μM, 7 μM, 15 μM, 30 μM, and 60 μM respectively). When the concentration of parthenolide was below 30 μM, there was no significant difference in cell viability compared with the absence of parthenolide, and the cell viability remained above 100% with a promoting effect on cell proliferation. When the concentration of parthenolide reached 60 μM, there was a significant difference compared with the concentration of parthenolide, and the cell viability reached 81.45%, showing an inhibitory effect on cells. In summary, 30 μM was selected as the maximum concentration of parthenolide for a series of cell experiments.

[0042] (2) GLP-1 concentration determination: Caco-2 cells were seeded at a density of 1×10 4Cells were cultured in a 96-well plate at a density of cells / mL and incubated for 24 h. Then, the cells were starved in DMEM without bovine serum albumin for 12 h. 100 μL of parthenolide solutions at different concentrations (0 μM, 3.75 μM, 7 μM, 15 μM, 30 μM respectively) were added to the 96-well plate. After incubation at 37 °C for 2 h, the supernatant was collected and stored at -80 °C for analysis. According to the manufacturer's protocol, GLP-1 levels were measured using a GLP-1 ELISA kit.

[0043] A series of non-toxic concentrations of parthenolide (0 μM, 3.75 μM, 7 μM, 15 μM, 30 μM respectively) were used to determine its effect on GLP-1 secretion by intestinal endocrine cells Caco-2. The results are as Figure 3 shown. As the concentration of parthenolide increased, GLP-1 secretion gradually increased; when the concentration of parthenolide reached 15 μM, the GLP-1 secretion was significantly different from that without parthenolide solution, indicating that parthenolide can be used as a GLP-1 receptor agonist to promote GLP-1 secretion by intestinal endocrine cells Caco-2.

[0044] The above examples show that parthenolide is a secretagogue with high efficiency in secreting GLP-1 and low side effects, which will be a promising direction for increasing endogenous GLP-1 levels, providing a good scientific basis for the development of effective products for weight intervention and maintaining healthy blood glucose levels in the future, and having great potential market value.

[0045] The above description of the embodiments is to enable those of ordinary skill in the art to understand and use the invention. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative efforts. Therefore, the present invention is not limited to the above embodiments, and the improvements and modifications made by those skilled in the art without departing from the scope of the present invention according to the interpretation of the present invention should be within the protection scope of the present invention.

Claims

1. Use of parthenolide and / or parthenolide derivatives in the preparation of a drug for preventing and / or treating diabetes, characterized in that: The parthenolide and / or parthenolide derivatives serve as GLP-1 receptor agonists.

2. Use of parthenolide and / or parthenolide derivatives according to claim 1 in the preparation of a drug for preventing and / or treating diabetes, characterized in that: The drug is a drug that promotes cells to secrete GLP-1, thereby promoting insulin secretion and reducing glucagon content.

3. Use of parthenolide and / or parthenolide derivatives according to claim 1 in the preparation of a drug for preventing and / or treating diabetes, characterized in that: The effective concentration of parthenolide and / or parthenolide derivatives in the medicine is 15-30 μM.

4. The use of parthenolide according to claim 3 in the preparation of a medicament for preventing and / or treating diabetes, characterized in that: The effective concentration of parthenolide and / or parthenolide derivatives in the drug is 15 μM.

5. A drug for preventing and / or treating diabetes, characterized in that: The medicine contains parthenolide and / or parthenolide derivatives.

6. Use of parthenolide and / or parthenolide derivatives in the preparation of weight intervention and / or blood sugar maintenance products, characterized in that: The parthenolide and / or parthenolide derivatives serve as GLP-1 receptor agonists.

7. Use of parthenolide and / or parthenolide derivatives according to claim 6 in the preparation of weight intervention and / or blood sugar maintenance products, characterized in that: The product is a product that inhibits gastric emptying and reduces food intake.

8. The use of parthenolide and / or parthenolide derivatives according to claim 6 in the preparation of weight intervention and / or blood sugar maintenance products, characterized in that: The effective concentration of parthenolide and / or parthenolide derivatives in the product is 15 to 30 μM.

9. The use of parthenolide and / or parthenolide derivatives according to claim 8 in the preparation of weight intervention and / or blood sugar maintenance products, characterized in that: The effective concentration of parthenolide and / or parthenolide derivatives in the product is 15 μM.

10. A product for weight intervention and / or blood sugar maintenance, characterized in that: The product contains parthenolide and / or parthenolide derivatives; The product is selected from food, cosmetics, health products or medicines.

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