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Anti-inflammatory tetrapeptide, extraction and separation method therefor and application of anti-inflammatory tetrapeptide in preparation of drugs for improving memory

A technology for improving memory and pro-inflammatory mediators, applied in the field of active peptides, to achieve the effect of inhibiting activity and alleviating brain neuroinflammation

Active Publication Date: 2020-06-09
SOUTH CHINA UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Conversely, the process will generate more ROS

Method used

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  • Anti-inflammatory tetrapeptide, extraction and separation method therefor and application of anti-inflammatory tetrapeptide in preparation of drugs for improving memory
  • Anti-inflammatory tetrapeptide, extraction and separation method therefor and application of anti-inflammatory tetrapeptide in preparation of drugs for improving memory
  • Anti-inflammatory tetrapeptide, extraction and separation method therefor and application of anti-inflammatory tetrapeptide in preparation of drugs for improving memory

Examples

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Embodiment 1

[0033] BV-2 cell culture

[0034] Fetal bovine serum (FBS) was added to the DMEM medium in an amount of 10% (v / v), and then a mixed double antibody composed of 100 U / mL penicillin and 100 μg / mL streptomycin was added. The cell culture incubator provides the required environment for the growth of BV-2 cells, and the conditions in the incubator are set at 37°C and contain 5% CO 2 . Change the medium and subculture in time according to the growth of the cells.

Embodiment 2

[0036] Synthesis of Gly-Val-Tyr-Tyr Polypeptide by Solid Phase Synthesis

[0037] Swell and wash the dichloro resin, remove the Fmoc protecting group, add amino acid to carry out condensation reaction, repeat the process of removal-protection-condensation until all amino acids are connected. Cut the resin to obtain the crude polypeptide Gly-Val-Tyr-Tyr, and purify it by reverse-phase high-performance liquid chromatography to obtain the pure polypeptide (>95%).

Embodiment 3

[0039] Separation and purification method of Gly-Val-Tyr-Tyr polypeptide

[0040] (1) Preparation of walnut protein powder: extract walnut protein from walnut dregs by alkali-dissolving and acid-precipitating methods. The specific process is as follows, after walnut meal is powdered, add water to dissolve (material ratio is 1:12) → adjust the pH value of the solution to 8.5 → stir for 2 hours → centrifuge (8000rpm, 15min, 4°C) → take the supernatant and adjust the pH value to 4.5 → stir 2h→centrifugation (8000rpm, 15min, 4°C)→precipitate dissolution→dialysis desalination for 2d (slag and water 1:5)→drying→walnut protein powder.

[0041] (2) Preparation of enzymatic hydrolyzate of walnut protein: Take 1 part of walnut protein powder and 8 parts of deionized water by weight and mix evenly, use two kinds of proteases (composite plant hydrolase and trypsin) to enzymolyze at 55 °C for 12 hours, compound plant The amounts of hydrolase and trypsin added were 1.0% and 1.0% (w / w), res...

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Abstract

The invention discloses an anti-inflammatory tetrapeptide, an extraction and separation method therefor and application of the anti-inflammatory tetrapeptide in preparation of drugs for improving memory. The anti-inflammatory tetrapeptide has an amino acid sequence of Gly-Val-Tyr-Tyr. The extraction and separation method for the anti-inflammatory tetrapeptide comprises the following steps: extracting walnut proteins from walnut meal through alkali-solution and acid-isolation, weighing the walnut proteins, adding deionized water into the weighed walnut proteins for mixing, carrying out enzymolysis, and taking supernatant, so as to obtain a walnut protein enzymolysis solution; collecting permeate with an ultrafiltration membrane with the molecular weight of 3kDa; and introducing a lower-molecular-weight component sample of the walnut protein enzymolysis solution to a gel filtration chromatographic column, carrying out eluting with deionized water, employing a detection wavelength of 220nm, collecting and merging a plurality of elution peak components, carrying out mass spectrometric detection on the component with relatively high anti-inflammatory activity, and confirming that the component contains the anti-inflammatory tetrapeptide disclosed by the invention. The anti-inflammatory tetrapeptide disclosed by the invention has relatively good inhibiting activity to LPS stimulatedBV-2 cell inflammation reactions and can be used for preparing memory improving drugs or healthy products.

Description

technical field [0001] The invention belongs to the field of active peptides, and in particular relates to an anti-inflammatory tetrapeptide, its extraction and separation method and its application in the preparation of memory-improving drugs. Background technique [0002] Inflammatory response refers to the complex biological response of the microcirculatory system caused by harmful stimuli, infection or trauma. Normally, inflammation is initiated by a series of soluble mediators (complement released by inflammation-related cells, chemokines, cytokines, and reactive oxygen species, etc.) and then the focus is eliminated. However, overexpression or chronic inflammation is an underlying cause of many diseases, including cancer, rheumatoid arthritis, chronic asthma, multiple sclerosis, obesity, autoimmune diseases, diabetes, inflammatory bowel disease, and cardiovascular disease, among others. In the brain, neuroinflammation has been implicated in the pathogenesis of neurode...

Claims

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Application Information

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IPC IPC(8): C07K5/103C07K1/04C07K1/06C07K1/34C12P21/06A61K38/07A61P29/00A61P25/00
CPCA61K38/00A61P25/00A61P29/00C07K5/1008C12P21/06
Inventor 苏国万赵谋明王曙光郑淋
Owner SOUTH CHINA UNIV OF TECH
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