Scorpion active polypeptide adp-7 and its application

A technology of ADP-7 and scorpion active peptides, which is applied in the fields of peptides, peptide sources, allergic diseases, etc., can solve the problems of toxic and side effects, strong effects, and failure to enter clinical trials, etc., and achieves small toxic and side effects, convenient operation, and drug significant effect

Active Publication Date: 2021-04-16
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can be seen from the latest review of new drug research on potassium ion channel Kv1.3 related peptides published by our team (Bingzheng Shen, Zhijian Cao, Wenxin Li, Jean-Marc Sabatier&Yingliang Wu. Treating autoimmune disorders with venomderived peptides, Expert Opinion on Biological Therapy, 17(9) : 1065-1075), candidate organic small molecule drugs almost all act on potassium channel Kv1.3 and other similar potassium channels, so they are likely to cause strong toxic side effects, which makes them unable to enter clinical trials at present

Method used

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  • Scorpion active polypeptide adp-7 and its application
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  • Scorpion active polypeptide adp-7 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] [Example 1] Purification and identification of scorpion active polypeptide ADP-7

[0026] The scorpion active peptide ADP-7 was submitted to Qiangyao Biotechnology Co., Ltd. (Suzhou, China) for linear peptide synthesis. On this basis, the 3 pairs of disulfide bonds in the polypeptide molecule are cyclized to form a polypeptide with a tertiary structure. The specific method is: dissolve the polypeptide in 0.1M Tris-HCl (pH=8.0) with a final concentration of 0.1mM (Huamei Bioengineering Co., Ltd.), after incubation at 23°C for 48 hours, it was separated by RP-HPLC, and the eluted peak containing the cyclized target polypeptide was collected ( figure 1 ).

[0027] Such as figure 1 The elution time of the reduced form of the scorpion active polypeptide ADP-7 is 19.5 minutes, while the elution time of the oxidized form of the scorpion active polypeptide ADP-7 is about 3 minutes later. The disulfide bond cyclized polypeptide ADP-7 was identified by mass spectrometry. Such...

Embodiment 2

[0028] [Example 2] Pharmacological activity analysis of scorpion active polypeptide ADP-7 on potassium channel Kv1.3

[0029] HEK-293T cells were cultured in DMEM medium containing 10% fetal bovine serum at 37°C, 5% CO 2 conditions, the potassium channel Kv1.3 and hERG recombinant plasmids were used 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 active polypeptide ADP-7 was measured and analyzed by using the 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 10kHz (Bessel), and the electrode impedance is 2-5MΩ. After forming a high-resistance (1-5GΩ) seal between the elect...

Embodiment 3

[0031][Example 3] Drug efficacy experiment of scorpion active polypeptide ADP-7 in the treatment of multiple sclerosis

[0032] Experimental animals: inbred female Wistar rats (6-8 weeks old, body weight 150±10g) and guinea pigs (300-400g purchased from the Experimental Animal Center of Wuhan University) were selected.

[0033] Main reagents: Freund's complete adjuvant (Gibcol / BRL), BCG, pertussis vaccine (Shanghai Institute of Biological Products), guinea pig MBP (Sigma).

[0034] Preparation of whole spinal cord homogenate-freund's complete adjuvant mixed emulsion (GPSCH-CFA): after the guinea pigs were sacrificed, the spinal cord was quickly taken out, and a 50% PBS homogenate was made with a sonicator (Sonics&Materials Inc, America). A certain amount of Freund's complete adjuvant (BCG 10mg / ml) was mixed and pumped with a syringe to form a water-in-oil emulsion.

[0035] Induction of EAE EAE model in Wistar rats: Intradermally inject 0.4ml GPSCH-CFA emulsion into the foot ...

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Abstract

The invention discloses a scorpion active polypeptide ADP-7 and an application thereof, belonging to the field of biotechnology. The scorpion active polypeptide designed in the present invention has the amino acid sequence shown in SEQ ID NO:1. The polypeptide has high selectivity and low toxicity as a potassium ion channel Kv1.3 blocker. The invention provides the application of the scorpion active polypeptide in medicine for treating or preventing diseases related to potassium channel Kv1.3.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to scorpion active polypeptide ADP-7 and its application. Background technique [0002] Autoimmune disease is a disease caused by the body's immune response to self-antigens, 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, including T cell-mediated rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus, Behcet's disease, autoimmune Thyroid disease and type I diabetes etc. In rheumatoid arthritis, the immune system attacks the patient's joints, and in multiple sclerosis, the myelin sheaths of nerve cells are attacked. 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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/435A61K38/17A61P37/02A61P19/02A61P25/00
CPCA61K38/00C07K14/43522
Inventor 吴英亮曹志贱
Owner WUHAN UNIV
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