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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  • 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

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Example Embodiment

[0032] Example 1

[0033] BV-2 cell culture

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

Example Embodiment

[0035] Example 2

[0036] Synthesis of Gly-Val-Tyr-Tyr peptides by solid phase synthesis

[0037] The dichloro resin is swelled and washed, the Fmoc protective group is removed, and amino acids are added for condensation reaction, and the removal-protection-condensation process is repeated until all amino acids are connected. The resin was cut to obtain the crude peptide Gly-Val-Tyr-Tyr, which was purified by reversed-phase high performance liquid chromatography to obtain the pure peptide (> 95%).

Example Embodiment

[0038] Example 3

[0039] Gly-Val-Tyr-Tyr polypeptide separation and purification method

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

[0041] (2) Preparation of walnut protease hydrolysate: Take 1 part of walnut protein powder and 8 parts of deionized water by weight and mix them evenly. Use two proteases (composite plant hydrolase and pancreatin) to hydrolyze at 55°C for 12 hours. The amount of hydrolase and pancreatin added was 1.0% and 1.0% (w / w), respectively. Subseque...

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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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