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Application of 3beta-acetyl oleanolic acid in preparing drug for treating autoimmune diseases

A technology for acetyl oleanolic acid and autoimmune diseases, applied in the field of biomedicine, can solve problems such as uncertainty, and achieve the effects of significantly inhibiting function, reducing the degree of kidney damage, and reducing the total number of

Inactive Publication Date: 2019-01-04
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no Th17 cell activity inhibitor based on the RORγt target for clinical use, and some of the inhibitors discovered are still in the preclinical research stage, and whether they can be drugged is still uncertain

Method used

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  • Application of 3beta-acetyl oleanolic acid in preparing drug for treating autoimmune diseases
  • Application of 3beta-acetyl oleanolic acid in preparing drug for treating autoimmune diseases
  • Application of 3beta-acetyl oleanolic acid in preparing drug for treating autoimmune diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 EC of 3β-acetyl oleanolic acid in RORγt--Jurkat cells 50 、CC 50 test

[0022] 1. The luciferase activity screening system based on transcription factor activity is constructed as follows:

[0023] Material:

[0024] The cell line Jurkat was preserved in our laboratory; the DH5α bacterial strain was donated by Professor Ma Runlin, Institute of Genetics, Chinese Academy of Sciences, Beijing; the restriction endonuclease was purchased from Fermentas Company of the United States; the DNA ligase was purchased from NEB Company of the United States; the reporter gene sequence IRES-GFP was invented Preserved by people themselves (other existing reporter gene sequences can also be used); reporter plasmid pGL4.31[luc2P / GAL4UAS / Hygro] plasmid and pBIND plasmid were purchased from Promega Company in the United States; DMEM medium, RPMI1640 medium and cultured cells were used Sodium pyruvate, glutamine, β-mercaptoethanol, non-essential amino acids, and double antibodies...

Embodiment 2

[0037] Example 2 Test of the inhibitory effect of 3β-acetyl oleanolic acid on Th17 cells, IL-17A and IL-17F

[0038] Through in vitro Th17 cell differentiation experiments, it was verified that 3β-acetyl oleanolic acid has the function of inhibiting Th17 differentiation. The inhibitory effect of 3β-acetyl oleanolic acid on Th17 cells, IL-17A, and IL-17F was studied by flow cytometry, and the test results were as follows: figure 2 A. figure 2 B. figure 2 C shown.

[0039] Experimental operation: The night before the experiment, a 12-well plate was coated with a PBS solution containing 5 μg / mL CD3 antibody and 1 μg / mL CD28 antibody, 0.5 mL per well, and coated overnight at 4°C. The experiment was carried out on the second day. First, the CD4+ T cells in the mouse spleen were sorted with Miltenyi magnetic beads, and the sorted cells were divided into 1×10 6 Resuspend the cells at a cell density of 1 / mL, add 1 mL of cell suspension to each well of the coated 12-well plate, ...

Embodiment 3

[0044] Example 3 3β-acetyl oleanolic acid inhibits serum anti-dsDNA in a mouse model of lupus nephritis and the therapeutic effect of nephritis

[0045] To establish the lupus nephritis model induced by pristane, a single injection of 500 μL pristane was administered intraperitoneally, and a single injection of 500 μL normal saline was used as the control. 3β-acetyl oleanolic acid is compared with solvent control and positive drug prednisone, and the medicine is dissolved in 25% dehydrated alcohol and 75% hydroxypropyl β-cyclodextrin (content is 30%), Oral administration of 100 μL was administered twice a week for 2 consecutive months. Then study the inhibitory effect of 3β-acetyl oleanolic acid on anti-dsDNA in mouse serum. The result is as image 3 A, 3B shown.

[0046] Such as image 3 As shown in A and 3B, 2 is the initial serum anti-dsDNA antibody concentration of 2-month-old mice before establishing the nephritis model, 6 is the serum anti-dsDNA antibody concentratio...

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Abstract

The invention relates to application of 3beta-acetyl oleanolic acid in preparing a drug for treating autoimmune diseases and belongs to the technical field of biomedicines. The 3beta-acetyl oleanolicacid can inhibit functions of a transcription factor RORgammat, differentiation of a Th17 cell, transcriptional expression of target genes IL-17A and IL-17F molecules of RORgammat in RNA level, secretion of an IL-17A cell factor and level of an anti-dsDNA antibody in blood serum, reduce the total quantity of IL-17A cells in spleen and reduce the renal degree of injury. The 3beta-acetyl oleanolic acid provided by the invention has an obvious function of inhibiting activity of the Th17 cell and provides a piloting structure for research and development of a new drug for treating autoimmune diseases.

Description

technical field [0001] The invention relates to the application of 3β-acetyl oleanolic acid in the preparation of medicines for treating autoimmune diseases, and belongs to the technical field of biomedicine. Background technique [0002] Autoimmune diseases affect approximately 5% of the population, and more than 70 human diseases, including psoriasis, multiple sclerosis, rheumatoid arthritis, asthma, and inflammatory bowel disease, are associated with autoimmune disorders. At present, the treatment of autoimmune diseases mainly relies on some non-selective immunosuppressants, which have limited curative effect and large side effects, so there is no very effective drug for the treatment of autoimmune diseases in clinical practice. Therefore, it is an urgent clinical need to develop novel autoimmune drugs with high efficacy and low side effects. Recent studies have shown that a new T cell subset, Th17 cells, is associated with the occurrence of human autoimmune diseases and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/56A61P37/02
CPCA61K31/56A61P37/02
Inventor 黄朝峰何细新柏川陈焕鹏周晓庆
Owner SUN YAT SEN UNIV
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