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Use of scorpion source immunoloregulation polypeptide in preparing medicine for treating rheumatoid arthritis

A rheumatoid and arthritis technology, applied in the field of scorpion-derived active polypeptide BmKTX in the treatment or prevention of rheumatoid arthritis, can solve the problems of poor potassium channel selectivity, toxic and side effects, etc., and achieve low toxic and side effects and specificity Strong, drug-like effect

Active Publication Date: 2011-08-17
WUHAN MORE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Poor selectivity of potassium channels with different roles and serious potential side effects have become the pathogenic shortcomings of small molecule drug development. This is mainly because the amino acid residue sequence of the Kv1.1-Kv1.7 target region of organic small molecule drug candidates is almost complete unanimous

Method used

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  • Use of scorpion source immunoloregulation polypeptide in preparing medicine for treating rheumatoid arthritis
  • Use of scorpion source immunoloregulation polypeptide in preparing medicine for treating rheumatoid arthritis
  • Use of scorpion source immunoloregulation polypeptide in preparing medicine for treating rheumatoid arthritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Expression and purification of recombinant scorpion-derived active polypeptide BmKTX

[0023]The expression vector plasmid pGEX-6p-1-BmKTX containing the scorpion source active polypeptide BmKTX gene (constructed by inserting the gene sequence encoding BmKTX into the expression vector pGEX-6p-1 purchased from Amersia Company, Biochem.J. 2000 346:805-809) was transformed into E. coli DE3. The transformed Escherichia coli was cultured after induction with IPTG, and the cells were collected, and the bacteria were disrupted by ultrasonic waves, and the fusion protein was obtained by GST affinity chromatography. The collected fusion protein solution is desalted and concentrated, digested with enterokinase (EK), and chromatographically separated to obtain the recombinant scorpion-derived active polypeptide BmKTX.

[0024] The BmKTX polypeptide is further separated by high-pressure liquid chromatography, and the GST protein is removed to obtain a chromatographicall...

Embodiment 2

[0025] Example 2: Analysis of the pharmacological activity of the recombinant scorpion-derived active polypeptide BmKTX on the potassium channel Kv1.3 target

[0026] COS-7 cells were cultured in DMEM medium containing 10% fetal bovine serum at 37°C and 5% CO2, and the potassium channel Kv1.3 recombinant plasmid was treated with Sofast TM The transfection kit was used for transfection, and the transfected cells were selectively cultured on 0.8mg / ml Geneticin medium. The pharmacological activity of the recombinant scorpion-derived active polypeptide BmKTX was determined and analyzed using a whole-cell patch clamp instrument (EPC-10 dual-channel patch clamp amplifier HEKA, Elektronik, Lambrecht, Germany). The setting of experimental parameters, data acquisition and application of stimuli were all controlled by Pulse software (Elektronik, Lambrecht, Germany). The filter of the instrument is set to 10 kHz (Bessel), and the electrode impedance is 2-5MΩ. After a high-resistance (1-...

Embodiment 3

[0027] Example 3: Use of the scorpion-derived active polypeptide BmKTX as a drug for treating or preventing rheumatoid arthritis.

[0028] Select 40 specific pathogen-free (SPF) grade inbred SD rats (Experimental Animal Center of Wuhan University), ♀, weighing 150±10g, and feed them adaptively for 1 week under clean environmental conditions, with a dose of 0.2ml / rat The pristane (pristane) was injected intradermally at the base of the tail of the rats in the experimental group. Symptoms of rheumatoid arthritis appeared after 2 weeks of feeding. Then the arthritis rats were randomly divided into: 10 model mice + normal saline (model negative control group); 10 model mice + methotrexate (model positive control group); 10 model mice + polypeptide drugs; another normal control group 10 (negative control group). Administration group according to 150μg·kg -1 The dose was injected subcutaneously, once a day in the morning; the model negative control group and the negative control ...

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Abstract

The invention discloses an application of scorpion source activity polypeptide BmKTX in preparing medicaments for curing or preventing rheumatoid arthritis. The scorpion source activity polypeptide BmKTX is produced and reconstructed by the gene project. The polypeptide medicament is characterized by: (1) strong particularity; the IC50 value of the scorpion source activity polypeptide BmKTX function medicament target potassium channel Kv1.3 is 102pM, and the scorpion source activity polypeptide BmKTX function medicament is the existing known polypeptide medicament with strong activity in the international world; (2) obvious medicament effect; the animal experiments prove that the recombination of the BmKTX polypeptide can cure rheumatoid arthritis of rats effectively; and (3) small toxic side effects; in animal experiments, the scorpion source activity polypeptide BmKTX does not cause obvious toxicity and side effects on experimental animals. The invention is simple and feasible in method and convenient in operation.

Description

technical field [0001] The invention belongs to the field of biotechnology, and more specifically relates to the use of a scorpion-derived active polypeptide BmKTX in medicine for treating or preventing rheumatoid arthritis. Background technique [0002] Autoimmune disease (autoimmune disease) is a disease caused by the body's immune response to self-antigens and resulting in damage to its own tissues, that is, the human immune system attacks its own tissues. About 5-8% of the world's population is threatened by more than 40 autoimmune diseases. These diseases involve one or more tissues and organs throughout the body, seriously affecting human health and life. For autoimmune diseases, there is currently no effective therapy, and the disease recurs. At present, the commonly used methods to fight autoimmune diseases, one is to use steroids to slow down the inflammatory response caused by the immune system attacking tissues, and the other is to use immunosuppressive drugs to...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/17A61P19/02A61P29/00
Inventor 李文鑫吴英亮曹志贱韩松
Owner WUHAN MORE BIOTECH
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