Enzymolysis polypeptide and application thereof in preparation of anti-leukemia drug

An enzymatic hydrolysis and total protein technology, applied in the field of peptides, can solve the problem of very few peptide researches

Active Publication Date: 2021-03-09
魏骏峰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004]Currently, the research on Schisandra chinensis mainly focuses on the small molecular compounds in Schisandra chinensis, and there are very few studies on the polypeptides contained in it

Method used

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  • Enzymolysis polypeptide and application thereof in preparation of anti-leukemia drug
  • Enzymolysis polypeptide and application thereof in preparation of anti-leukemia drug
  • Enzymolysis polypeptide and application thereof in preparation of anti-leukemia drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] First, experimental materials

[0019] Pulling the dry ripe fruit of North Schisando over 60 mesh sieves, spare.

[0020] Pure water is pure water for Wahaha built; alkaline protease is purchased from Novi.

[0021] Second, the experimental method

[0022] Step S1: Alkali acidic acid extraction total protein

[0023] The dry ripe fruit of northeast is 1 kg first with 10 l of anhydrous ethanol heat reflux for 3 h, and the residual solvent is waved. Add 10 L of pure water, adjusting pH to 10.5 with a 10% sodium hydroxide solution, and stirring at normal temperature for 6 h. The filtrate was extracted to collect the filtrate, then centrifuged at 4000 r / min for 20 min, taking the supernatant, adjusted pH to 4.5 with a 10% hydrochloric acid solution, and centrifuged at 4000 r / min for 20 min, collected, and frozen. protein.

[0024] Step S2: Protease enzymatic

[0025] Approximately a solution of fresh water was used to dissolve a solution of 0.1 mg / ml using pure water, and...

Embodiment 2

[0030] First, experimental materials

[0031] Acute lymphocytic leukemia Jurkat cells were purchased from ATCC.

[0032] Fetal bovine serum, RP-Mi1640 culture liquid purchase from Hyclone.

[0033] The sputum enzymatic polypeptide was prepared in accordance with an example method.

[0034] Second, the experimental method

[0035] 1, cell culture

[0036] Acute lymphocyte leukemia Jurkat cells were placed at 37 ° C with RP-Mi1640 culture fluid containing 10% fetal bovine serum and 1% double-resistant, volume fraction of 5% CO 2 , Conventional cultivation in the incubator of saturated humidity, replacement of 1 culture fluid per 2D.

[0037] 2, MTT test assay cell proliferation activity

[0038] Take a few long-term JURKAT cell digestion resuspended into a cell suspension, inoculated in a 96-well plate, 1 × 10 per hole 4 Cell, 200 μL, 37 ° C, 5% CO 2 Conditions were cultured for 24 hours, and the complete medium containing 10,20 μg / ml of spicy polypeptide was added, and a pharmace...

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Abstract

The invention discloses an enzymolysis polypeptide and application thereof in preparation of an anti-leukemia drug. The enzymolysis polypeptide is prepared by the following method of firstly, extracting schisandra chinensis total protein from dry mature fruits of schisandra chinensis by using an alkali extraction and acid precipitation method, and then, carrying out enzymolysis on the schisandra chinensis total protein by using alkaline protease; and finally, collecting the schisandra chinensis enzymolysis polypeptide with the molecular weight between 5000u and 7000u by using ultrafiltration separation. The schisandra chinensis enzymolysis polypeptide provided by the invention has obvious inhibitory activity on acute lymphocytic leukemia Jurkat cells, and can be used for preparing drugs for resisting acute lymphocytic leukemia.

Description

Technical field [0001] The present invention belongs to the field of polypeptides, and involves an enzymatic hydrolysis polypeptide and an application, and more particularly to an enzymatic hydrolysis polypeptide and a use of drugs for preparing anti-leukemia. Background technique [0002] Acute lymphocytic leukemia belongs to invasive blood system malignant tumors, accounting for about 75% of leukemia in childhood, and is also common in adults. It is characterized in that the T-progenitor cell abnormally cloned proliferation, resulting in a clinical symptoms such as anemia, anemia, fever, liver spleen, joint pain and systemic lymphatic node, and a large-scale lymph node. Acute lymphocyte leukemia incidence increased year by year, seriously threatening people's lives and health. Finding to develop effective chemotherapeutic drugs is an important way to overcome acute lymphocyte leukemia. [0003] Schisandra is a perennial plant plant for Mulanko, and the mature fruit is dry after...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C07K1/14C07K1/34C12P21/06A61K38/16A61K38/01A61P35/02
CPCC07K14/415C12P21/06A61P35/02A61K38/00
Inventor 郭守山其他发明人请求不公开姓名
Owner 魏骏峰
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