Centipede toxin anti-tumour active polypeptide

A technology with anti-tumor and anti-tumor effects, applied in the direction of anti-tumor drugs, peptides, peptide/protein components, etc., can solve the problems of less research on single components of centipedes and retention of anti-tumor components in centipedes

Inactive Publication Date: 2010-12-01
CHINA PHARM UNIV
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Problems solved by technology

But so far, the research on the anti-tumor components of centipede mostly stays at the level o

Method used

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  • Centipede toxin anti-tumour active polypeptide
  • Centipede toxin anti-tumour active polypeptide

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Experimental program
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Embodiment Construction

[0012] 1. Preparation of centipede toxin anti-tumor active polypeptide

[0013] a. Take 1,000 centipedes from Chuzhou, Anhui Province, use electrical stimulation to extract poison, and freeze-dry. Weigh 0.4g centipede toxin, dissolve in 20ml PBS solution, and centrifuge at 10000r / min for 5min.

[0014] b. The above solution was ultrafiltered with a 10000Da ultrafiltration tube, and centrifuged at 7000r / min for 15min.

[0015] c. Collect the filtrate, use C 18 - HPLC column separation and purification, the column height is 25cm, and the diameter is 0.46cm.

[0016] After equilibrating with 20ml of 5% acetonitrile (containing 0.05% TFA), load 500μl of the filtrate, first elute with 20ml of 5% acetonitrile (containing 0.05% TFA), and then use 30ml of 95% acetonitrile (containing 0.05% TFA) 0-60% gradient elution with a flow rate of 1 ml / min, the active peak 7 was collected and freeze-dried.

[0017] The following drawings are used to illustrate specific embodiments of the pre...

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Abstract

The invention discloses polypeptide separated from centipede toxin, and determination on anti-tumour activity thereof is carried out. Ultrafilteration and reversed-phase chromatography separation methods are adopted to separate and purify one polypeptide composed of twelve amino acids from centipede toxin, the sequence thereof detected by applying edman degradation method is Phe-Thr-Gly-Gly-Asp-Glu-Ser-Arg-Ile-Gln-Glu-Gly, and the molecular weight thereof measured by applying mass-spectrometric method is 1296.05Da. The anticancer cell viability of the polypeptide is determined by applying MTT method. The polypeptide in the invention has anti-tumour activity, has inhibition action on propagation and growth of human umbilical vein endothelial cell, is in dose-response relationship and can be applied to treatment and auxiliary treatment of various cancers.

Description

1. Technical field [0001] The invention belongs to the technical field of polypeptide medicine in biochemistry. 2. Background technology [0002] Centipede is an animal of the family Centipede of Arthropoda Phylum Chiliopoda Plastic Order. Centipede has been used as medicine in China for more than 2,000 years. It was first recorded in "Shen Nong's Materia Medica". , Indications for stroke, epilepsy, tetanus, whooping cough, tuberculosis, burns, sores and swelling. Now pharmacological research shows that centipede also has antitumor effect, antispasmodic effect, antifungal effect and the effect of treating diabetes. [0003] Although the distribution range of centipede is wide, the research on centipede venom is far behind the venom of other poisonous animals such as snakes and scorpions, and there are few reports about centipede toxin. Centipede toxins contain substances such as serotonin, histamine, acid and alkaline phosphatases, cardiotoxic proteins, and neurotransmitte...

Claims

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Application Information

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IPC IPC(8): C07K7/08A61K38/10A61P35/00
Inventor 孔毅惠晶曹艳华赖伊丽邵妤杨天雨
Owner CHINA PHARM UNIV
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