Application of JAM-A (junctional adhesion molecules A) genes or protein in genetic recombination drugs for treating rheumatoid arthritis

A JAM-A, gene recombination technology, applied in the direction of drug combination, microbial determination/examination, antipyretic drugs, etc., can solve the problems of lack of clinical treatment methods, complex pathogenesis, adverse reactions of patients, etc., and achieve simple production process, The effect of short production cycle and easy expansion of large-scale production

Inactive Publication Date: 2018-09-14
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The occurrence and development of rheumatoid arthritis is an autoimmune process involving the lymphatic system and various cytokines. Its pathogenesis is complex and has not been fully clarified yet, and there is also a lack of effective clinical treatment methods
At present, the commonly used drugs for the clinical treatment of rheumatoid arthritis are divided into four categories, namely nonsteroidal anti-inflammatory drugs (nonsteroidal anti-inflammatory drugs, NSAIDs), immunosuppressants, glucocorticoids and herbal medicines, such as methotrexate (methotrexate), azathioprine, cyclophosphamide, etc., but long-term use of these drugs will cause adverse reactions in many patients, resulting in intolerance and discontinuation

Method used

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  • Application of JAM-A (junctional adhesion molecules A) genes or protein in genetic recombination drugs for treating rheumatoid arthritis
  • Application of JAM-A (junctional adhesion molecules A) genes or protein in genetic recombination drugs for treating rheumatoid arthritis
  • Application of JAM-A (junctional adhesion molecules A) genes or protein in genetic recombination drugs for treating rheumatoid arthritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Example 1: Expression of JAM-A protein in peripheral blood lymphocytes of healthy people and RA patients.

[0014] 1. Collection and grouping of blood samples.

[0015] The peripheral blood sample collected in the present invention informs volunteers and patients with rheumatoid arthritis of the experimental purpose before blood collection and signs an informed consent. review.

[0016] The present invention collected peripheral blood from 48 patients with rheumatoid arthritis (rheumatoid arthritis group, RA), all from the Department of Rheumatology and Immunology, the First Affiliated Hospital of Kunming Medical University. The peripheral blood of 37 healthy adult volunteers (normal group, CON) was collected from the First Affiliated Hospital of Kunming Medical University and volunteers of Yunnan University. Each patient and volunteer was asked about the medical history by two attending physicians in the department of rheumatology and immunology and underwent a speci...

Embodiment 2

[0019] Example 2: Effect of JAM-A expression on lymphocyte migration ability.

[0020] 1. Common rheumatoid arthritis treatment drugs can effectively increase the expression of junctional adhesion molecule A.

[0021] Lymphocytes treated with 0.01 μg / mL methotrexate for 5 days were collected, total RNA was extracted and extracted using Takara’s PrimeScript RT reagent Kit with gDNA Eraser (Perfect Real Time) (Takara, Code No.DRR047A) kit. RNA is reverse transcribed into cDNA. The expression of junctional adhesion molecule A was analyzed by fluorescent quantitative PCR with the following primers:

[0022] Human junction adhesion molecule A (301-409, 109bp):

[0023] Forward: 5'- CGGGAAGACACTGGGACATA- 3';

[0024] Reverse: 5'-CTGTAGGCTTGGATGGAGGC-3';

[0025] Human β-actin (939-1143, 205bp):

[0026] Forward: 5'- TGACGTGGACATCCGCAAAG-3';

[0027] Reverse: 5'-CTGGAAGGTGGACAGCGAGG-3'.

[0028] The amplification program was pre-denaturation at 94°C for 3 min; 36 cycles of 30 ...

Embodiment 3

[0031] Example 3: Treatment of Collagen-Induced Arthritis (CIA) Mice with Adhesion Molecule A Linked Neutralizing Antibody J10.4.

[0032] 1. Grouping of experimental mice.

[0033] Select 6-8 weeks and C57BL / 6 female mice with a body weight of 22-26 g (Beijing Weitong Lihua Company, clean grade), and intradermally immunize bovine type II collagen at the base of the tail to establish a CIA model, and the time of mouse immunization Defined as Day 0 (Day 0). 8 normal control groups (Control group, CON) and 24 diseased mice were randomly divided into three groups. The first group was the CIA model group without any treatment; the second group was the junction adhesion molecule A specific antibody J10.4 To treat the CIA model group, at the peak of CIA, that is, 20 days after collagen immunization, 1 mg / kg of J10.4 neutralizing antibody was injected into the tail vein every other day for 10 consecutive days. The third group is the IgG negative control group, the immunization time...

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Abstract

The invention identifies junctional adhesion molecules A (JAM-A) highly expressed in peripheral blood lymphocytes of patients suffering from rheumatoid arthritis, proves that the highly-expressed JAM-A can effectively reduce migration of lymphocytes to peripheral bone joints, inhibit immunoreactions and joint inflammation, reduce bone destruction degree and relieve RA (rheumatoid arthritis) conditions, and relates to products in the fields of genetic engineering and biotechnological pharmaceutics. The production process of drugs made from the JAM-A is simple, mass production expansion is facilitated, technological conditions are easy to control, and production cycle is short. The invention mainly relates to application of JAM-A genes or protein encoded by the JAM-A genes, in particular toapplication of the JAM-A genes or the protein in genetic recombination drugs for treating the rheumatoid arthritis. The JAM-A genes are recombined with genetic engineering carriers to prepare the genetic recombination drugs capable of improving expression of the JAM-A, so that the rheumatoid arthritis can be treated effectively. The drugs can be in the form of injections or oral solutions.

Description

technical field [0001] The invention belongs to products in the field of genetic engineering and biotechnology pharmacy, and mainly relates to the application of junctional adhesion molecule A (Junctional Adhesion Molecules A, JAM-A) gene or protein, and more specifically relates to the preparation of junctional adhesion molecule A gene or protein The application of gene recombination drugs in the treatment of rheumatoid arthritis (Rheumatoid Arthritis, RA), that is, to recombine the adhesion molecule A gene with other gene carriers to prepare a drug that can effectively control rheumatoid arthritis, and its drug dosage form can be an injection type or oral form. Background technique [0002] Rheumatoid arthritis (Rheumatoid Arthritis, RA) is an autoimmune disorder characterized by chronic synovial inflammation, which eventually leads to cartilage resorption, bone destruction and bone fibrosis. Rheumatoid arthritis is prevalent in the world The prevalence rate is 0.42-0.54%,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883G01N33/68A61K39/395A61P19/02A61P29/00
CPCA61K2039/505A61P19/02A61P29/00C12Q1/6883C12Q2600/158G01N33/6893G01N2800/102
Inventor 丁磊李梅章王维
Owner YUNNAN UNIV
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