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Preparation method of natural scorpion venom polypeptide and application of natural scorpion venom polypeptide to bone marrow haematopoietic functional protection

A technology of hematopoietic function and bone marrow, which is applied in the field of application in the preparation of drugs for the protection of bone marrow hematopoietic function

Inactive Publication Date: 2015-06-24
孔天翰 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are many researches on the cell biological effect and mechanism of action of scorpion venom and its toxin (single-component pure product) in inhibiting cell growth at home and abroad. etc.) There are no similar reports

Method used

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  • Preparation method of natural scorpion venom polypeptide and application of natural scorpion venom polypeptide to bone marrow haematopoietic functional protection
  • Preparation method of natural scorpion venom polypeptide and application of natural scorpion venom polypeptide to bone marrow haematopoietic functional protection
  • Preparation method of natural scorpion venom polypeptide and application of natural scorpion venom polypeptide to bone marrow haematopoietic functional protection

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preparation example Construction

[0029] The object of the present invention is also to provide a kind of preparation method of natural scorpion venom polypeptide, comprising the following steps:

[0030] (1) Collection of East Asian scorpion scorpion venom

[0031] In this embodiment, the adult East Asian scorpion is taken from the poisonous species breeding base of Dongguan Zhongke Bio-Pharmaceutical Co., Ltd., and the fresh venom is extracted by electric stimulation. Toxin collection instrument: JL-D multi-purpose instrument for pharmacological and physiological experiments (produced by Shanghai Yilian Science and Education Equipment Co., Ltd.); instrument parameters: continuous pulse 25-30Hz, wave width 0.5-0.7ms, voltage 6-8 volts; operation method: use One electrode clamped one of the scorpion's tentacles, and then a metal clip was used to clamp the 5th segment of the scorpion's rear abdomen. The other electrode was used to lightly touch the metal clip, and the electrical stimulation was stopped after th...

Embodiment 1

[0044] Example 1: BmKpp prolongs the survival of mice after radiation

[0045] C57BL / 6 mice received 7.5Gy total body irradiation (TBI) (6M high-energy linear accelerator, dose rate 300cGy / min, SIMENS PRIMUS). C57BL / 6 mice receiving 7.5Gy X-ray radiation were randomly divided into radiation control group (IR + group) and BmKpp treatment group (IR + +BmKpp group: BmKpp 2.63 μg / kg). Fifteen mice in each group were intraperitoneally injected with BmKpp or normal saline 2 hours before TBI and every other day after radiation until 14 days after TBI. Another 15 normal mice were taken as the non-irradiated control group (IR - group), and saline was injected intraperitoneally at the same time point. After TBI, the mice were weighed every day, the hair, stool, and mental state of the mice were observed, and the symptoms, signs, and death numbers and time of the animals were recorded until 30 days after radiation. Survival experiments showed that IR + Group C57BL / 6 mice all died w...

Embodiment 2

[0050] Example 2: BmKpp promotes the recovery of mouse hematopoietic function after radiotherapy

[0051] Mice were irradiated into IR + Group and IR + In the +BmKpp group, the animal administration method was the same as above. Before TBI and 5d, 7d, 14d, and 28d after TBI, blood was collected from the inner canthus of the mouse eyes, anticoagulated with EDTA, and the whole blood cells were routinely checked by F-820 animal blood cell analyzer (Sysmex, Japan) within 1 hour to analyze peripheral blood leukocytes The total number (WBC), the number of neutrophils (Number of neutrophils), the number of platelets (PLT number) and hemoglobin (HB). IR + Group and IR + The C57BL / 6 mice in the +BmKpp group were killed by dislocation, the tibia and femur were taken, the bone marrow fluid was washed out with IMDM, the supernatant was removed after centrifugation, the cells were resuspended in IMDM containing 3% FBS, and about 6ml of red blood cell lysate was added to lyse the red bl...

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Abstract

The invention discloses a natural scorpion venom polypeptide. The natural scorpion venom polypeptide is a novel protein separated from scorpion venom of Buthus martensii Karsch and has a novel medicinal purpose on the aspect of bone marrow haematopoietic functional protection. A preparation method of the natural scorpion venom polypeptide comprises the following steps of (1) collecting the scorpion venom of Buthus martensii Karsch; (2) separating SVPIV; (3) purifying BmKpp; (4) measuring the molecular weight and amino acid sequence of of BmKpp; and (5) measuring the bone marrow haematopoietic functional protection effect of BmKpp. BmKpp is a novel polypeptide from scorpion venom of Buthus martensii Karsch and has a remarkable bone marrow haematopoietic functional protection effect. BmKpp is single in component, precise in molecular weight, clear in total amino acid sequence, remarkable in cell proliferation promoting effect as well as standard and controllable in quality in the purification process so as to be beneficial to large-scale production. BmKpp has remarkable advantages and prospects on the aspects of bone marrow haematopoietic functional protection and development of drugs for preventing / protecting cell stress injury (radiotherapy, chemotherapy and the like).

Description

technical field [0001] The invention relates to a natural scorpion venom polypeptide, its preparation method and application. Specifically, it relates to a new natural polypeptide isolated from the scorpion venom of Buthus martensii Karsch, and its application in the preparation of drugs with protective effect on bone marrow hematopoietic function. technical background [0002] There are 6 families, 70 genera, and more than 800 species of scorpions in the world. In China, the East Asian scorpion is the main species. The latter has historically been known both at home and abroad as one of the traditional and precious Chinese medicines in my country. Scorpion venom is a poisonous secretion secreted from the venom sac of the scorpion scorpion. It is the main effective part of the scorpion to treat diseases. Its curative effect on tumors, pain, and cardiovascular diseases is several times stronger than that of the scorpion. The pharmacological effect of scorpion venom fighting ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07K1/16A61K38/17A61P7/06
CPCC07K14/43522A61K38/00
Inventor 董伟华孔天翰郭奇峰王燕王彩霞刘四红邱轶芳蒋丽苑李婷邢百倩
Owner 孔天翰
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