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The application of mud snail polypeptide in anti-prostate cancer

A technology for anti-prostate cancer and mud snail polypeptide, which is applied in the preparation methods of peptides, peptides, anti-tumor drugs, etc., can solve the problems of large toxic and side reactions, poor selectivity, etc., and achieve the effects of low cost, mild reaction and simple process

Active Publication Date: 2020-11-27
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Malignant tumors are a type of disease that seriously threatens human health and life, and the incidence rate is getting higher and higher. Although the existing chemotherapy drugs have certain curative effects on most tumors, they have poor selectivity, large toxic and side effects, and drug resistance. The problem is still very obvious

Method used

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  • The application of mud snail polypeptide in anti-prostate cancer
  • The application of mud snail polypeptide in anti-prostate cancer
  • The application of mud snail polypeptide in anti-prostate cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: the preparation of mud snail polypeptide

[0024] (1.1) Raw material handling

[0025] Take fresh mud snails, shell the mud snails, drain and homogenize for later use.

[0026] (1.2) Snail enzymatic hydrolysis process

[0027] Use a high-speed tissue grinder to mash the mud snail tissue and homogenize it, accurately weigh the homogenate solution, adjust the pH value with 0.1mol / L hydrochloric acid solution and 0.1mol / L NaOH solution, add trypsin for several hours, and enzymolysis The conditions are as follows: enzymolysis temperature 45°C, pH 8.7, solid-liquid ratio 1:4, enzymolysis time 8h, enzyme addition 0.48%. Inactivate the enzyme at 100°C for 15 minutes, centrifuge at 4°C for 15 minutes (10000r / min), and concentrate the supernatant for later use.

Embodiment 2

[0028] Example 2: Culture and passage of DU-145 cells

[0029] (2.1) Cell Recovery

[0030] Take out the stored DU-145 cell strain from the liquid nitrogen tank, quickly put it into a 37°C constant temperature water bath for melting, enter the sterile studio for operation after melting, and suck the cell strain into the centrifuge tube with a sterilized straw , add 2mL of F12 nutrient solution or RPMI1640 nutrient solution, centrifuge at 1000rpm for 10min, remove the supernatant, add 4ml of nutrient solution, and repeatedly pipette to make the cells into single cells. Then use a pipette to evenly suck the cells into two 25mL culture flasks, put 5% CO 2 , Culture in a constant temperature incubator at 37°C, and change the medium the next day to discard unattached dead cells.

[0031] (2.2) Cell culture

[0032] Human prostate cancer cells DU-145 were inoculated into F12 and 1640 nutrient solutions containing 10% fetal bovine serum (volume fraction) FBS and double antibodies ...

Embodiment 3

[0037] : Embodiment 3: the separation and purification of enzymolysis crude extract

[0038] First, the ultrafiltration method is used to separate the crude extract from the enzymatic hydrolysis of the mud snail polypeptide. The ultrafiltration membranes are respectively 10kd, 5kd, and 3kd, and the three components with molecular weights of 10-5kd, 5-3kd and less than 3kd are intercepted respectively. After freeze-drying The concentration of 20 mg / mL was prepared respectively, and the inhibition rate of each component on prostate cancer DU145 was determined by MTT method. Select the component with the highest activity, choose Sephadex G-25 gel chromatography, equilibrate with deionized water after loading the column, the concentration is 50mg / mL, the sample volume is 4mL, the speed is 3ml / min, and the mobile phase is super Pure water, 280nm UV detection. The elution peaks were collected and lyophilized into powder, and the inhibition rate of DU-145 cell proliferation was dete...

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Abstract

The invention relates to application of bullacta exarata polypeptides in resisting prostatic cancer, in particular to application of bullacta exarata polypeptides in medicine resisting prostatic cancer DU-145 cells. A bullacta exarata polypeptide crude extract is obtained through a trypsin enzymolysis method, all bullacta exarata polypeptide components are obtained through an ultrafiltration method, a gel column separation method, a reverse-phase high performance liquid chromatogram technology and other separation and purification means, the inhibiting capacity of each separated component on proliferation of prostatic cancer DU-145 cells is studied, and the study shows that all the components can remarkably inhibit proliferation of prostatic cancer DU-145 cells. The raw material cost is low, extraction conditions are mild, extracted bullacta exarata polypeptides can remarkably inhibit proliferation of prostatic cancer DU-145 cells, and theoretical bases are provided for developing prostatic cancer resisting medicine with bullacta exarata polypeptides as raw materials.

Description

technical field [0001] The invention relates to the application of the mud snail enzymolysis polypeptide, in particular to the application of the mud snail polypeptide in anti-prostate cancer. Background technique [0002] Mud snail (Bullacta exarata), commonly known as "spitting iron", "yellow mud snail", "plum snail", etc., belongs to the mollusc phylum, gastropoda, hindbranch subclass, cephalopod, Adi snail family, mud snail The genus is a special product of seawater and brackish water on the west coast of the Pacific Ocean. It is composed of shells and soft bodies. It is rich in protein, calcium, phosphorus, iron and vitamins. It is rich in polypeptide substances and is widely distributed on both sides of the East China Sea and the Yellow Sea in my country. The Yangtze River is a common aquatic product. [0003] Malignant tumors are a type of disease that seriously threatens human health and life, and the incidence rate is getting higher and higher. Although the existing ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/01C12P21/06C07K1/36C07K1/34C07K1/16C07K1/20A61P35/00
Inventor 闻正顺马剑茵曹玉昊林焕乐胡俊峰
Owner ZHEJIANG OCEAN UNIV
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