Oligopeptide and active peptide composition and application of oligopeptide and active peptide composition in preparation of product with anti-inflammatory effect
An anti-inflammatory and active peptide technology, applied in the field of biomedicine, can solve the problems of chronic inflammation and cell damage in the skin, and achieve the effects of convenient operation, enhanced activity, and simple preparation process
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Embodiment 1
[0033] Example 1 The separation of bifidobacterium oligopeptide-α and Saccharomyces cerevisiae cyclic peptide-β
[0034] Mix 50g of bifidobacteria with 500mL of ethyl acetate and 2000mL of water evenly, ultrasonicate for 20min, extract the water phase layer after standing, and repeat the extraction process several times, vacuum and freeze-dry the water phase layer, and then carry out preparative HPLC Prepare and isolate to obtain bifidobacterium oligopeptide-α (HT-1).
[0035] Mix 50g of brewer's yeast with 500mL of ethyl acetate and 2000mL of water, ultrasonically for 20min, extract the water phase layer after standing, repeat the extraction process several times, vacuum and freeze-dry the water phase layer, and then carry out preparative HPLC Prepare and isolate to obtain Saccharomyces cerevisiae cyclic peptide-β (DF-1).
[0036] The preparation conditions of the above-mentioned preparative HPLC are: 0.1% trifluoroacetic acid aqueous solution is used as mobile phase A, and ...
Embodiment 2
[0037] Example 2 Chemical Synthesis of Bifidobacterium Oligopeptide-α (HT-1) and Saccharomyces Saccharomyces Cyclopeptide-β (DF-1)
[0038](1) Take 100mg of Fmoc-Tyr Wang Resin and place it in a solid-phase synthesis tube, add N,N-dimethylformamide (DMF) and let it stand for 30 minutes to fully swell the resin, filter off the solvent, and then add piperidine DMF solution, and the solvent was filtered off after shaking. Dissolve Fmoc-Ser-OH, 1-hydroxybenzotriazole, O-benzotriazole-tetramethyluronium hexafluorophosphate in DMF, add N,N-diisopropylethylamine and mix well After being protected from light, it has been activated, and added to the resin, blown and stirred with nitrogen at 25°C for 2 hours, filtered with suction, washed with DMF and dichloromethane in sequence, and dried the solvent. Repeat the above steps, add the activated Fmoc-Leu-OH, Fmoc-His-OH, Fmoc-Leu-OH, Fmoc-His-OH, Fmoc-Gly-OH to the resin in sequence, blow and stir with nitrogen at 25°C, After the reacti...
experiment example
[0044] In order to evaluate the biological activity of the bifidobacterium oligopeptide-α (HT-1) and Saccharomyces cerevisiae cyclic peptide-β (DF-1) of the present invention and their combinations, the following effect examples were carried out.
[0045] The immortalized keratinocyte line HaCaT cells were cultured in a cell incubator at 37°C, 5% CO 2 (DMEM medium). The concentration of LPS was 1 μM to stimulate for 12 hours. After the end, the cells were washed with PBS, the PBS was discarded, and DMEM medium or medium containing HT-1 or DF-1 (20 μM) was added to continue culturing for 24 hours. The CCK8 method was used to detect cell viability, and the cells and culture fluid were collected.
[0046] In the test of the effect of HT-1 or DF-1 on the viability of HaCaT cells, the experiment was divided into 4 groups: normal control group (Control); LPS group; HT-1 20μM group (LPS+HT-1 20μM); DF-1 20 μM group (LPS+DF-1 20 μM). Each treatment condition had 3 replicate wells, ...
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