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Application of polypeptide in preparation of drugs for treatment and/or prevention of rheumatoid arthritis

A rheumatoid and arthritis technology, applied in the application field of polypeptides in the preparation of drugs for the treatment or prevention of rheumatoid arthritis, can solve the problems of lack of TRB3 protein inhibitors, etc., and achieve the effect of improving inflammatory response and animal joint inflammation

Active Publication Date: 2015-07-01
胡卓伟
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The technical problem to be solved by the present invention is to provide a polypeptide that can specifically bind to TRB3 and its use in the preparation of drugs for preventing and / or treating rheumatoid arthritis in view of the current lack of effective TRB3 protein inhibitors. Applications

Method used

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  • Application of polypeptide in preparation of drugs for treatment and/or prevention of rheumatoid arthritis
  • Application of polypeptide in preparation of drugs for treatment and/or prevention of rheumatoid arthritis
  • Application of polypeptide in preparation of drugs for treatment and/or prevention of rheumatoid arthritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1 rheumatoid arthritis in vitro model preparation

[0049] 1.1 Model Construction

[0050] 1) Collection of synovial or bone samples from patients with rheumatoid arthritis undergoing knee or wrist synovectomy or joint replacement.

[0051] 2) Under sterile conditions, the sample was cut into 2 cubic millimeter pieces and divided into several parts and placed in two 24-well plates.

[0052] 3) Samples were prepared using MEM (Gibco, Grand Island, NY, USA) complete medium (containing 2mM L-glutamine, 100U / ml penicillin, 50mg / ml gentamicin, 20mM N-2-hydroxyethylpiperazine -N'-2-ethanesulfonic acid buffer solution and 1% fetal bovine serum) 2 mL per well, 37°C, 5% CO 2 , cultivate overnight at 95% humidity.

[0053] 4) The shredded tissue pieces are cultured in the culture medium, and the inflammatory cells and other cells in the tissue pieces enter the culture medium as the tool cells for the next day's drug treatment experiment.

[0054] 1.2 Therapeutic Ac...

Embodiment 2

[0066] The co-immunoprecipitation method in Example 2 verifies the interaction between protein p62 and protein TRB3 in chondrocytes.

[0067] 2.1 Reagents

[0068] 1) Lysis solution A: Lysis solution A: 0.6057g Tris base, 1.7532g NaCl, 0.1017g MgCl 2 ·6H 2 O, 0.0742g EDTA, 10mL glycerin, 10mL10%NP-40, add deionized water to 150mL, adjust the pH value to 7.6 with HCl, adjust the volume to 191mL, mix well, filter through a 0.45μm membrane filter, and store at 4°C.

[0069] 2) Lysis solution B: 200 μL 2M β-glycerol phosphate, 4 mL 2.5M NaF, 2 mL 100 mM NaVO3, 2 mL 100 mM PMSF (phenylmethylsulfonyl fluoride), 200 μL 1M DTT (dithiothreitol), 1 mg / mL Leupeptin (leupeptin) , Pepstatin (pepsin inhibitor), and Aprotinin (aprotinin) are 200 μL each, and the total volume is 9 mL. The mother liquor was stored at -20°C. Before use, thaw the mother solution of each component in the B solution, add it to the A solution according to the above composition ratio and mix well, and obtain the...

Embodiment 3

[0078] 3.1 Materials

[0079] 1. Animals

[0080] SPF grade SD rats, female, body weight 180-200g. Purchased from Beijing Weitong Lihua Experimental Animal Co., Ltd., the laboratory controls the temperature at 23-25°C, the humidity at 60-70%, and regularly exhausts and ventilates.

[0081] 2. Drugs and reagents

[0082] Methotrexate tablets, produced by Huayi Pharmaceutical Factory of Shanghai Pharmaceutical (Group) Co., Ltd., 1 mg / tablet. Batch number: 120815. The immune grade bovine type II collagen is the product of American Chondrex Company (Catalog No. 20022), 2mg / ml, dissolved in 0.05M acetic acid solution. Freund's incomplete adjuvant (Freund's incomplete adjuvant) is a product of Sigma Company in the United States (10 mL / cartridge). Rat IgG, IL1-β, TNF-α enzyme chain immunoassay kits, products of American R&D Company (Research&Diagnostics System).

[0083] 3.2 Method

[0084] 1. Model construction

[0085] Take 2mg / ml type II collagen dissolved in 0.05M acetic ...

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PUM

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Abstract

The invention discloses an application of a polypeptide or polypeptide derivatives capable of specific combination of TRB3 in preparation of drugs for treatment and / or prevention of rheumatoid arthritis; the polypeptide has the amino acid sequence being any one of sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3; the polypeptide derivatives are chimeric peptides formed by connecting the polypeptide with cell membrane cell-penetrating peptides. The polypeptide or the polypeptide derivatives can be specifically combined with the TRB3 so as to block the interaction between the TRB3 and P62 proteins; and the peptide fragment derivatives Pep2-A1, Pep2-A2 and Pep2-A3 can treat or prevent various symptoms caused by rheumatoid arthritis.

Description

technical field [0001] The invention relates to the application of a polypeptide and its derivatives in the preparation of medicines for treating and / or preventing rheumatoid arthritis. Background technique [0002] Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by chronic erosive arthritis. Rheumatoid arthritis is characterized by synovitis, and the resulting destruction of articular cartilage and bone, eventually leading to joint deformity. Without formal treatment, about 75% of patients will become disabled within 3 years. Rheumatoid arthritis is distributed all over the world, and the prevalence rate in different populations is 0.18% to 1.07%. There are certain racial differences in its incidence. Indians are higher than Caucasians, and Caucasians are higher than Asians . The total number of patients in my country exceeds 5 million. Rheumatoid arthritis can occur in all ages, and the peak age is around 30 to 50 years old. Generally, women ar...

Claims

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Application Information

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IPC IPC(8): A61K38/08A61K47/48A61K45/00A61P19/02A61P29/00
Inventor 胡卓伟花芳孙巍
Owner 胡卓伟
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