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Scorpion venom heat-resistant synthetic peptide and application thereof

A technology for synthesizing peptides and scorpion venom, which is applied to medical preparations with no active ingredients, medical preparations containing active ingredients, peptides, etc., and can solve the problem of low peptide extraction rate from scorpion venom heat-resistant peptide extracts

Active Publication Date: 2016-12-14
上海万锦医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the rate of obtaining peptides from scorpion venom heat-resistant peptide extracts is low, so obtaining chemically synthesized scorpion venom heat-resistant peptides is a key link in determining whether this original research and development achievement can be transformed into R&D and industrialized production

Method used

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  • Scorpion venom heat-resistant synthetic peptide and application thereof
  • Scorpion venom heat-resistant synthetic peptide and application thereof
  • Scorpion venom heat-resistant synthetic peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Amino Acid Sequence Determination of Heat-resistant Peptide from Scorpion Venom

[0036] (1) Redissolve and centrifuge the freeze-dried powder of BmK scorpion venom from Yichang, Henan Province with three-distilled water, and take the supernatant, which is the BmK scorpion venom extract; pack it in a heat-resistant plastic tube with a cover, and place it in a constant temperature water bath In the process, heat at 100°C for 4 hours, take it out, cool down to room temperature naturally, and then centrifuge at a high speed to get the supernatant, which is the extract of heat-resistant components of scorpion venom; The filter tube is used to perform centrifugal ultrafiltration on the heat-resistant component extract of scorpion venom, and take the upper bath solution to obtain the heat-resistant polypeptide extract of scorpion venom. Take the supernatant by centrifugation, which is the extract of heat-resistant components of scorpion venom. Use centrifugal ultrafiltration ...

Embodiment 2

[0046] Purification of Synthetic Scorpion Venom Heat-resistant Polypeptide by High Performance Liquid Chromatography (HPLC)

[0047] In the process of peptide synthesis, some heteropeptides with structures similar to the target peptide will be produced, such as diastereoisomers due to amino acid racemization, missing peptides due to unlinked partial amino acids, and fragmentation due to peptide bond cleavage peptide etc. Therefore purify with RP-HPLC:

[0048] Sample purification conditions:

[0049] Structure: KE-15

[0050] Serial number: 0200046

[0051] Batch number: P160418-CQ455216

[0052] Column: 4.6mm*250mm, Inertsil ODS-SP

[0053] Solution A: Acetonitrile with 0.1% trifluoroacetic acid

[0054] Solution B: water containing 0.1% trifluoroacetic acid

[0055]

[0056] Flow rate: 1.0ml / min

[0057] Wavelength: 214nm

[0058] Injection volume: 10μl

Embodiment 3

[0060] Using Morris water maze test to detect the effect of SVHRP on learning and memory in AD mice

[0061] The water maze was used to detect behavioral changes. The positioning navigation experiment lasted 5 days, trained 4 times a day, and recorded the time when the mice found the platform (the platform was placed in the first quadrant), that is, the escape latency period, and allowed the mice to stay on the platform for 20 seconds; The mouse did not find the platform within 60s, and the escape latency was recorded as 60s. In the space search experiment, after the 4th training on the 5th day of the positioning navigation test, the platform was removed, and then the mice were put into the pool at the water entry point in the third quadrant to start the test. Calculate the swimming distance and swimming time percentage of mice in the target quadrant (quadrant 1), and the total time is 60s. The number of times the mice crossed the hidden platform was detected.

[0062] The M...

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Abstract

The invention belongs to the field of research and development of polypeptide medicines and discloses a scorpion venom heat-resistant synthetic peptide and application thereof. An amino acid sequence of an SVHRP (scorpion venom heat-resistant peptide) is detected out from BmK (Buthus martensii Karsch) which is a traditional Chinese medicine; according to animal experiment verification, a scorpion venom heat-resistant peptide extract liquid sample has pharmacological activity in prevention and treatment of intractable epilepsy, Parkinson's disease and Alzheimer's disease, the sample is subjected to LaGm composite and repeated fast magnetic separation prior to nanoLC-ESI-MS (nano-liter reversed-phase chromatography and electrospray ionization mass spectrometry) integrated mass spectrometry parallel experiment to detect out a polypeptide sequence formed by 15 amino acid residues. The scorpion venom heat-resistant synthetic peptide is prepared by solid-phase chemical synthesis, chromatography purification and mass spectrometry identification. An amino acid sequence of the scorpion venom heat-resistant synthetic peptide is as shown in SEQ ID NO.1 and keeps pharmacological activity and safety of the scorpion venom heat-resistant peptide, and the scorpion venom heat-resistant peptide also has a characteristic of biological activity in promotion of reverse differentiation of neuroglial cells into neural stem cells.

Description

technical field [0001] The invention belongs to the field of polypeptide drug research and development, and in particular relates to a heat-resistant peptide obtained from the venom of the traditional Chinese medicine Scorpion scorpion scorpion and its application in the treatment of epilepsy, Alzheimer's disease and Parkinson's disease. Background technique [0002] The clinical symptoms of Parkinson's disease (PD) are caused by the severe reduction of striatal DA neurotransmitter levels due to damage to dopamine (DA) neurons in the substantia nigra of the midbrain. The theory of oxidative stress in PD was put forward because DA oxidative stress-free radical theory is an important mechanism of DA neuron damage in the substantia nigra compacta (NCs) of the midbrain, and antioxidant therapy is currently recognized as an effective treatment plan. [0003] Buthus martensii Karsch (BmK), also known as Buthus martensii Karsch (BmK), is a traditional Chinese medicine. Modern medic...

Claims

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Application Information

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IPC IPC(8): C07K7/08A61K38/10C07K1/16C07K1/04A61P25/08A61P25/28A61P25/16
CPCC07K7/08A61K38/00A61P25/16A61P25/28A61P25/08A61K2300/00A61K47/6415C12N2310/3513C07K14/43522A61K47/6817A61P29/00C07K14/43518A61P25/02C07K14/43504A61P25/04A61K38/02
Inventor 赵杰李韶张万琴
Owner 上海万锦医药科技有限公司
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