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Scorpion venom heat-resisting polypeptide, and preparing method and applications thereof

A technology of heat-resistant polypeptide and scorpion venom, applied in the field of heat-resistant polypeptide and preparation, can solve the problems of unremoved toxic components, animal death, and toxicity restricting clinical application, etc.

Inactive Publication Date: 2015-02-11
张万琴
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The toxicity of scorpion venom limits its clinical application
In 2001, the patent CN1324621 disclosed that a safe and effective heat-resistant component was isolated from BmK scorpion venom, but the application found that when its dose was increased to 55.5 mg / kg, it could still cause all the experimental animals to die. Invention finds reason for heat-resistant toxic components not yet removed

Method used

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  • Scorpion venom heat-resisting polypeptide, and preparing method and applications thereof
  • Scorpion venom heat-resisting polypeptide, and preparing method and applications thereof
  • Scorpion venom heat-resisting polypeptide, and preparing method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Fresh scorpion venom extracted from BmK in Yichang, Henan, and stored at -20°C after freeze-drying.

[0026] 2. Dissolve the freeze-dried powder of BmK scorpion venom in three-distilled water at a concentration of 50 mg / ml, centrifuge at 4500 rpm / min at 4°C for 15 minutes, and take the supernatant scorpion venom stock solution after centrifugation;

[0027] 3. Dispense the supernatant (1) with a 50ml sterile, high-temperature-resistant plastic tube with a cover; after drilling the plastic tube cover, cover the plastic tube and screw it tightly; place it in a constant temperature water bath for continuous For 4 hours, heat at 100°C with water; after 4h, take it out, cool down to room temperature naturally, and centrifuge at 7500rpm / min at 4°C for 15min; take the centrifuged supernatant (2), and then use 19000rpm / min, 4°C, high-speed Centrifuge for 40 minutes; take the supernatant (3) after high-speed centrifugation, which is the heat-resistant component of scorpion ve...

Embodiment 2

[0031] MALDI-TOF detection method and instrument parameters of heat-resistant polypeptide in scorpion venom.

[0032] 1. Reagents and Instruments

[0033] Water (Milli Q), acetonitrile (CAN, Fisher Scientific), trifluoroacetic acid (TFA, Merck)

[0034] MALDI TOF-Target (MTP 384, Bruker Daltonics)

[0035] Bruker Daltonics Autoflex (Bruker Daltonics Billerica, MA, USA)

[0036] 2. Experimental steps

[0037] 1) Sample processing

[0038] Wash and dissolve a little sample with 10 microliters of TA solution, take the supernatant of the solution and directly point it on the sample target, evaporate to dry at room temperature, absorb 0.6 microliter of matrix solution to cover the sample and calibration standard, and evaporate to dry at room temperature

[0039] 2) On-board testing

[0040] Instrument Operating Conditions

[0041] Delayed extraction time: (Delayed extraction) 190ns

[0042] Detection mode: Line mode / positive ion mode

[0043] Laser frequency: (Laser frequen...

Embodiment 3

[0051] Scorpion venom thermostable polypeptide increases the migration of stem cells in the SGZ region of the hippocampus, see Figure 7 , the number of PSA-NCAM immunoreactive positive cells increased, the grayish white is PSA-NCAM, and the dark gray is DAPI (nuclei). a, NS normal control group; b, single drug (scorpion venom heat-stable polypeptide 50 μg / kgBW) group; c, NMDA injury model group; d, NMDA injury model group + scorpion venom heat-stable polypeptide treatment group; e, MPTP PD model Group; f, MPTP PD model group + scorpion venom heat-resistant polypeptide. The results showed that the red PSA-NCAM in the treatment group all increased, indicating that the heat-resistant polypeptide of scorpion venom has a significant effect on the generation of endogenous neural stem cells in the neurogenesis area of ​​the brain.

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Abstract

The invention provides a scorpion venom heat-resisting polypeptide, and a preparing method and applications thereof, and belongs to the field of polypeptide and preparation thereof. The scorpion venom heat-resisting polypeptide which is safer is obtained by removing heat-resisting and heat-labile toxic components mainly from the scorpion venom of buthus martensii kirsch (BmK). The polypeptide is used for preventing and treating intractable epilepsy, Parkinson's disease (PD), Alzheimer's disease (AD), and other diseases, has a common function target, and has respective special medicine effects. Preparation of the polypeptide is stable in process, simple and controllable.

Description

technical field [0001] The invention belongs to the field of polypeptide and its preparation, in particular to a heat-resistant polypeptide, its preparation method and application. Background technique [0002] Scorpions are one of the oldest organisms, with an evolutionary history of 440 million years. Scorpion venom (Scorpion venom, SV) is the venom released by the scorpion venom gland when the scorpion preys and is provoked. [0003] According to clinical reports, since ancient times, scorpion or scorpion venom has been used as medicine to relieve convulsions and relieve wind, attack poison and dissipate stagnation, search wind and dredge collaterals, etc. Modern medical research has proved that the toxic components of scorpion venom are composed of neurotoxicity and cytotoxicity, and its neurotoxins are divided into long-chain and short-chain toxins, which have been used as tool drugs for the study of ion channels. In 1988, Zhou Xinhua of Shenyang Pharmaceutical Univer...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435A61K38/17A61P25/08A61P25/16A61P25/28
CPCC07K14/43522A61K38/00
Inventor 张万琴
Owner 张万琴
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