Cosmetic or pharmaceutical product containing active polypeptide and preparation method thereof
A technology of active peptides and drugs, which is applied in the field of drugs and cosmetics, can solve the problem of the accumulation and increase of ROS in the anti-oxidation defense mechanism, and achieve good application prospects, application value, and good anti-aging effects
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Embodiment 1
[0028] The separation and screening and identification of embodiment 1 active peptide
[0029] Take 10g of Baotou fish skin, soak it with 10 times the volume of 0.2% sodium hydroxide solution and stir it constantly, rinse it with running water until neutral after 50 minutes; then soak it with 0.2% sulfuric acid solution of 7 times the volume and keep stirring it for 40 minutes Finally, rinse with flowing water until neutral; finally add 10 times of distilled water to homogenate the fish skin, stir and extract with 50% hot water for 12 hours, and centrifuge (5000r / min, 20min) to collect the supernatant.
[0030] The above supernatant was first hydrolyzed under the optimal conditions of protease at pH 8.0, hydrolysis time 5h, enzyme-to-substrate ratio 5%, and hydrolysis temperature 35°C, then added alkaline protease for hydrolysis time 4.5h, pH 9.0 , Enzyme-to-substrate ratio of 5% and hydrolysis temperature of 55% for secondary hydrolysis, the prepared hydrolyzate was concentra...
Embodiment 2
[0034] Embodiment 2 Active peptide modification
[0035] The active peptide Ac-PYEMTCVMMWRYYFMWD was coupled with the transdermal peptide DNVFIGKNSSSYG at the C-terminus, and the conjugate Ac-PYEMTCVMMWRYYFMWDDNVFIGKNSSSYG was obtained by chemical synthesis. Weigh 200mg of the polypeptide and dissolve it in 50ml of buffer (0.1M PB, 0.15M NaCl) to prepare a 4mg / ml conjugate polypeptide solution.
[0036] Add 162.5 mg of lecithin, 25 mg of sodium cholate and 7 mL of a mixture of methanol and chloroform (V (methanol): V (chloroform) = 1:1) into a 50 mL round bottom flask; Sonicate in an ultrasonic instrument for 10min to disperse the solute (power is 50W); place the flask containing the ultrasonically dispersed solution in a rotary evaporator, and obtain a phospholipid film at 37°C under vacuum conditions; add to the flask containing the phospholipid film 15mL of the prepared conjugate polypeptide solution, put the flask in a shaker, and shake at 30°C for 3h to obtain a hydrated...
Embodiment 3
[0038] Embodiment 3 transdermal effect verification
[0039] Male SD rats were anesthetized with 1mL 20% ugline and killed by neck dislocation. The hair on their abdomen (3cm×3cm, length×width) was cut off with scissors, and the depilatory cream was applied. After 7min, the depilatory cream was removed. , wash the depilated skin 4 times with 37°C physiological saline. The mouse skin was removed with razor blades and tweezers and washed 4 times with normal saline to peel off the fat tissue on the skin, and the treated skin was placed in normal saline and placed in a refrigerator at 4°C for later use. The treated skin was fixed in the middle of the Franz tank, 5mL of normal saline and the magnetic rotor were added to the receiving tank (lower tank) of the Franz tank, and the air in the lower tank of the normal saline was removed by ultrasonication for 5 minutes, and then placed in a transdermal absorption instrument middle. Turn on the water circulation system of the transderm...
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