Composite peptide with anti-inflammatory and soothing effects, composition and application thereof

Abstract

The invention relates to a composite peptide with anti-inflammatory and soothing effects, a composition and application thereof, and belongs to the technical field of polypeptides. The invention relates to a composite peptide with anti-inflammatory and soothing effects. The composite peptide is prepared from the following components: tripeptide-1, acetyl dipeptide-1, palmitoyl tetrapeptide-7 and palmitoyl tripeptide-8, the mass ratio of the tripeptide-1 to the acetyl dipeptide-1 to the palmitoyl tetrapeptide-7 to the palmitoyl tripeptide-8 is (0.5 to 1.5) to (0.6 to 1.5) to (1 to 2) to (0.5 to 1.5). Experiments prove that the composite peptide has good anti-inflammatory, soothing and anti-wrinkle effects.

Description

Technical Field

[0001] The present invention belongs to the technical field of polypeptides and relates to a composite peptide with anti-inflammatory and soothing effects, a composition and applications thereof. Background Art

[0002] Cosmetics have become an indispensable part of our daily lives, especially for women, and the cosmetics market, especially products based on natural ingredients, is growing rapidly with the huge consumer demand for improved appearance. In particular, the development of new cosmetic formulations based on bioactive compounds with therapeutic and protective functions (such as antioxidants, proteins, peptides and growth factors) has expanded rapidly, and these bioactive compounds can provide significant effects such as whitening, moisturizing and anti-inflammatory effects on human skin.

[0003] Sensitive skin is a condition characterised by symptoms such as tightness, stinging, burning or itching, triggered by stimuli that do not normally produce unpleasant sensations, such as cold, heat, sunlight, pollution, cosmetics or humidity. The skin may also experience redness, dryness and flaking, but these symptoms are not usually present. Sensitive skin is thought to affect 71% of adults, and epidemiological studies have also shown that symptoms are more common on the face.

[0004] Most of the cosmetics for people with sensitive skin on the market now contain plant extracts with anti-allergic effects, but the mechanism of action of plant extracts is still unclear, the product efficacy is claimed to be single, and it may cause allergies, itching and other symptoms in people with sensitive skin. In addition, the plant extraction process is complicated and costly, which further limits its application. In the face of the above problems and the demand for anti-aging effects for people with sensitive skin, a product that can both relieve allergies and resist aging is urgently needed in the current market. As an active substance with multiple physiological functions, peptides have received more and more attention in the pharmaceutical, food, beauty and cosmetics industries, and are increasingly used in cosmetics. Some peptides with soothing effects can improve common problems of sensitive skin, such as neuritis, itching, tingling, desquamation, etc.

[0005] In the prior art invention, for the anti-allergic effect, peptide components are widely used, which are expensive. Moreover, the combination of peptides cannot take into account multiple effects, and there are many oil-soluble components, so more co-solvents need to be added, which causes skin sensitization. Summary of the invention

[0006] The main purpose of the present invention is to provide a composite peptide with anti-inflammatory and soothing effects and a composition containing the composite peptide. The composite peptide and the composition can be used to prepare cosmetics with anti-inflammatory and soothing effects.

[0007] The present invention adopts the following technical solutions to achieve the above purpose: A complex peptide with anti-inflammatory and soothing effects, mainly containing the following ingredients: tripeptide-1, acetyl dipeptide-1, palmitoyl tetrapeptide-7, and palmitoyl tripeptide-8.

[0008] Furthermore, the mass ratio of the tripeptide-1, acetyl dipeptide-1, palmitoyl tetrapeptide-7 and palmitoyl tripeptide-8 is (0.5-1.5):(0.6-1.5):(1-2):(0.5-1.5).

[0009] Furthermore, in the composite peptide, the mass concentration of tripeptide-1 is 20000 ppm, the mass concentration of acetyl dipeptide-1 is 3000 ppm, the mass concentration of palmitoyl tetrapeptide-7 is 50 ppm, and the mass concentration of palmitoyl tripeptide-8 is 350 ppm.

[0010] The present invention provides a specific application of the composite peptide, that is, the composite peptide can be used to prepare cosmetics with anti-inflammatory and soothing effects.

[0011] A composition containing the above-mentioned composite peptide, mainly comprising the following components by mass fraction: 0.8-1.2% tripeptide-1, 0.8-1.5% acetyl dipeptide-1, 1.0-1.5% palmitoyl tetrapeptide-7, 0.9-1.3% palmitoyl tripeptide-8, 3-6% glycerin, 3-8% Jerusalem artichoke polysaccharide, 0.5-1.5% PEG hydrogenated castor oil, 0.5-1.0% phospholipids, 0.1-0.14% vitamin E, and the balance is purified water.

[0012] Furthermore, the preparation method of the Jerusalem artichoke polysaccharide is: Step a, adding water to Jerusalem artichoke, adjusting the pH to 4.5-5.0, and refluxing extraction at 60-80° C. to obtain an extract; Step b, adjusting the pH of the extract obtained in step a to 7.0-7.5, extracting with ethyl acetate, taking the lower layer of solution, removing the residual ethyl acetate, concentrating at 60-80° C., and using the concentrate for later use; Step c, separating the concentrated solution obtained in step b by using a silica gel column, using a mixed solution of n-butanol-water-formic acid as an elution solvent, collecting the eluate, removing n-butanol from the eluate and concentrating it, adjusting the pH to 6.5-7.0, adding glucose oxidase and β-glucanase, performing enzymolysis at 35-40° C. for 4-6 hours, adding oryzanol and heating to 80-90° C. to inactivate the enzyme, adjusting the alcohol content of the enzymolysis solution to 20%, 30%, 50%, and 70% by using anhydrous ethanol in sequence, standing it for 4-6 hours after each adjustment of the alcohol content, centrifuging it, and taking the precipitate to obtain Jerusalem artichoke polysaccharide.

[0013] Furthermore, in the mixed solution of n-butanol-water-formic acid in step c of the above-mentioned method for preparing Jerusalem artichoke polysaccharide, the volume ratio of n-butanol, water and formic acid is (0.2-0.25):10:(0.06-0.08).

[0014] Furthermore, in step c of the Jerusalem artichoke polysaccharide preparation method, the amount of glucose oxidase added is 0.3-0.35 g / L, and the amount of β-glucanase added is 1-1.5 g / L.

[0015] Furthermore, in step c of the Jerusalem artichoke polysaccharide preparation method, the enzyme activity of glucose oxidase is 2000u / g, and the enzyme activity of β-glucanase is 20000u / g.

[0016] Furthermore, the amount of oryzanol added in step c of the above-mentioned Jerusalem artichoke polysaccharide preparation method is 5-8 g / L.

[0017] The present invention provides a method for preparing the above-mentioned composite peptide-containing composition, which specifically comprises the following steps: Step 1: Add palmitoyl tripeptide-8, palmitoyl tetrapeptide-7, vitamin E, and PEG hydrogenated castor oil into a container, heat to 60°C and stir until dissolved to prepare the oil phase.

[0018] Step 2: Add tripeptide-1, acetyl dipeptide-1, glycerol and water into a container and stir at room temperature to dissolve to prepare an aqueous phase.

[0019] Step 3: Add the water phase to the oil phase, stir evenly, add polysaccharide and phospholipid, and adjust the pH value to 5.0 to obtain the composition.

[0020] The present invention provides a specific use of the above-mentioned composite peptide composition, that is, the composition can be used to prepare cosmetics with anti-inflammatory and soothing effects, for example, it can be used alone as an essence or added with other ingredients to make cosmetics such as face cream, eye cream, facial mask, etc.

[0021] The present invention has the following beneficial effects: 1. The present invention uses tripeptide-1, acetyl dipeptide-1, palmitoyl tetrapeptide-7, and palmitoyl tripeptide-8 for effective compounding. Acetyl dipeptide-1 can stimulate the production of endorphins, release them around the nerve endings of the skin, reduce pain and burning sensations; inhibit the production of CGRP and SP that cause microvascular dilation and plasma extravasation, control the continuous vasodilation phenomenon, and relieve and improve skin sensitivity. Tripeptide-1 can promote cells to produce collagen, increase skin elasticity; repair damaged cells; inhibit pro-inflammatory factors, and regulate sensitive skin. Palmitoyl tetrapeptide-7 can improve uneven skin color, fade skin fine lines; improve skin elasticity, smooth and firm; reduce keratinocytes secrete interleukin IL-6, play an anti-inflammatory role; repair skin damage and stimulate the regeneration of collagen fibers in the dermis. Palmitoyl tripeptide-8 can reduce the release of Substance P and pro-inflammatory cytokines; reduce vasodilation, edema and erythema; prevent and reduce irritation, maintain and restore the normal sensitivity threshold of the skin.

[0022] The present invention uses less polypeptide components, and achieves both anti-allergic and anti-aging effects while having low components. At the same time, the important anti-allergic polypeptide acetyl dipeptide-1 cetyl ester is upgraded to acetyl dipeptide-1, which is water-soluble and does not require a solvent for dissolution in the cosmetic formula. The formula is simple and the cost is lower.

[0023] 2. In the composition containing the composite peptide of the present invention, vitamin E and Jerusalem artichoke polysaccharide are creatively added. The addition of these two ingredients can effectively ensure the stability of the composite peptide in different environments and ensure its sustained long-lasting biological activity. At the same time, it also helps to dissolve the composite peptide without adding co-solvents with sensitization risks. It is suitable for people with different skin types, especially those with sensitive skin, and ensures green safety. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 : ·OH removal rate at different times under room temperature; Figure 2 : O at different time under room temperature 2- Clearance rate; Figure 3 : ·OH removal rate at different times under UV conditions; Figure 4 : O at different times under UV conditions 2- Clearance rate. DETAILED DESCRIPTION

[0025] The present invention is further illustrated below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. After reading the present invention, modifications to various equivalent forms of the present invention by those skilled in the art fall within the scope of protection of the claims of this application. The mass concentration of tripeptide-1 used in the following specific implementation is 20000 ppm, the mass concentration of acetyl dipeptide-1 is 3000 ppm, the mass concentration of palmitoyl tetrapeptide-7 is 50 ppm, and the mass concentration of palmitoyl tripeptide-8 is 350 ppm; the enzymes used in the implementation are all existing commercially available products.

[0026] Example 1 Composite peptide A complex peptide with anti-inflammatory and soothing effects, mainly containing the following ingredients: 0.5kg tripeptide-1, 1.5kg acetyl dipeptide-1, 1kg palmitoyl tetrapeptide-7, 0.5kg palmitoyl tripeptide-8.

[0027] Example 2 Composite peptide A complex peptide with anti-inflammatory and soothing effects, mainly containing the following ingredients: 1.5kg tripeptide-1, 0.6kg acetyl dipeptide-1, 2kg palmitoyl tetrapeptide-7, 1.5kg palmitoyl tripeptide-8.

[0028] Example 3 Composite peptide A complex peptide with anti-inflammatory and soothing effects, mainly containing the following ingredients: 1kg tripeptide-1, 1kg acetyl dipeptide-1, 1.5kg palmitoyl tetrapeptide-7, 1kg palmitoyl tripeptide-8.

[0029] Example 4 Composition containing complex peptides and essence prepared therefrom Preparation of Jerusalem artichoke polysaccharide: Step a, adding 10 L of water to 1 kg of Jerusalem artichoke, adjusting the pH to 5.0, refluxing and extracting twice at 80° C., filtering, and combining the two filtrates to obtain an extract; Step b, adjusting the pH of the extract obtained in step a to 7.5, extracting with ethyl acetate, taking the lower layer of solution, removing the residual ethyl acetate, concentrating at 80° C., and using the concentrated solution for later use; Step c, loading the concentrated solution obtained in step b onto a silica gel column, using a mixed solution of n-butanol: water: formic acid = 0.2:10:0.06 by volume as an elution solvent, collecting the eluate, removing n-butanol from the eluate and concentrating it, adjusting the pH to 7.0, adding 0.35 g / L of glucose oxidase (enzyme activity of 2000 u / g) and 1 g / L of β-glucanase (enzyme activity of 20000 u / g), performing enzymolysis at 40° C. for 4 h, adding 8 g / L of oryzanol and heating to 80° C. to inactivate the enzyme, adjusting the alcohol content of the enzymolysis solution to 20%, 30%, 50%, and 70% with anhydrous ethanol in turn, standing for 6 h after each adjustment of the alcohol content, centrifuging, and taking the precipitate to obtain Jerusalem artichoke polysaccharide.

[0030] Prepare the composition and essence containing complex peptides according to the following ratio and method (1 kg)

[0031] Step 1: Add palmitoyl tripeptide-8, palmitoyl tetrapeptide-7, vitamin E, and PEG hydrogenated castor oil into a container, heat to 60°C and stir until dissolved to prepare the oil phase.

[0032] Step 2: Add tripeptide-1, acetyl dipeptide-1, glycerol and water into a container and stir at room temperature to dissolve to prepare an aqueous phase.

[0033] Step 3: Add the water phase to the oil phase, stir evenly, add Jerusalem artichoke polysaccharide and phospholipids, and use acetic acid and triethanolamine to adjust the pH value to 5.0.

[0034] Step 4: Filter (0.22μm filter membrane) and package in a GMP workshop or clean bench to obtain the essence.

[0035] Example 5, Preparation of Jerusalem artichoke polysaccharide: Step a, adding 10 L of water to 1 kg of Jerusalem artichoke, adjusting the pH to 4.5, refluxing and extracting twice at 60° C., filtering, and combining the two filtrates to obtain an extract; Step b, adjusting the pH of the extract obtained in step a to 7.0, extracting with ethyl acetate, taking the lower layer of solution, removing the residual ethyl acetate, concentrating at 60° C., and using the concentrate for later use; Step c, loading the concentrated solution obtained in step b onto a silica gel column, using a mixed solution of n-butanol: water: formic acid = 0.25:10:0.08 by volume as an elution solvent, collecting the eluate, removing n-butanol from the eluate and concentrating it, adjusting the pH to 6.5, adding 0.3 g / L of glucose oxidase (enzyme activity of 2000 u / g) and 1.5 g / L of β-glucanase (enzyme activity of 20000 u / g), performing enzymolysis at 35° C. for 6 hours, adding 5 g / L of oryzanol and heating to 90° C. to inactivate the enzyme, adjusting the alcohol content of the enzymolysis solution to 20%, 30%, 50%, and 70% with anhydrous ethanol in turn, standing for 4 hours after each adjustment of the alcohol content, centrifuging, and taking the precipitate to obtain Jerusalem artichoke polysaccharide.

[0036] Prepare the composition and essence containing complex peptides according to the following ratio and method (1 kg)

[0037] Step 1: Add palmitoyl tripeptide-8, palmitoyl tetrapeptide-7, vitamin E, and PEG hydrogenated castor oil into a container, heat to 60°C and stir until dissolved to prepare the oil phase.

[0038] Step 2: Add tripeptide-1, acetyl dipeptide-1, glycerol and water into a container and stir at room temperature to dissolve to prepare an aqueous phase.

[0039] Step 3: Add the water phase to the oil phase, stir evenly, add Jerusalem artichoke polysaccharide and phospholipids, and use acetic acid and triethanolamine to adjust the pH value to 5.0.

[0040] Step 4: Filter (0.22μm filter membrane) and package in a GMP workshop or clean bench to obtain the essence.

[0041] Example 6, Preparation of Jerusalem artichoke polysaccharide: Step a, adding 10 L of water to 1 kg of Jerusalem artichoke, adjusting the pH to 5.0, refluxing and extracting twice at 70° C., filtering, and combining the two filtrates to obtain an extract; Step b, adjusting the pH of the extract obtained in step a to 7.0, extracting with ethyl acetate, taking the lower layer of solution, removing the residual ethyl acetate, concentrating at 70° C., and using the concentrate for later use; Step c, loading the concentrated solution obtained in step b onto a silica gel column, using a mixed solution of n-butanol: water: formic acid = 0.23:10:0.07 by volume as an elution solvent, collecting the eluate, removing n-butanol from the eluate and concentrating it, adjusting the pH to 6.5, adding 0.33 g / L of glucose oxidase (enzyme activity of 2000 u / g) and 1.3 g / L of β-glucanase (enzyme activity of 20000 u / g), performing enzymolysis at 40° C. for 5 h, adding 7 g / L of oryzanol and heating to 80° C. to inactivate the enzyme, adjusting the alcohol content of the enzymolysis solution to 20%, 30%, 50%, and 70% with anhydrous ethanol in turn, standing for 5 h after each adjustment of the alcohol content, centrifuging, and taking the precipitate to obtain Jerusalem artichoke polysaccharide.

[0042] Prepare the composition and essence containing complex peptides according to the following ratio and method (1 kg)

[0043] Step 1: Add palmitoyl tripeptide-8, palmitoyl tetrapeptide-7, vitamin E, and PEG hydrogenated castor oil into a container, heat to 60°C and stir until dissolved to prepare the oil phase.

[0044] Step 2: Add tripeptide-1, acetyl dipeptide-1, glycerol and water into a container and stir at room temperature to dissolve to prepare an aqueous phase.

[0045] Step 3: Add the water phase to the oil phase, stir evenly, add Jerusalem artichoke polysaccharide and phospholipids, and use acetic acid and triethanolamine to adjust the pH value to 5.0.

[0046] Step 4: Filter (0.22μm filter membrane) and package in a GMP workshop or clean bench to obtain the essence.

[0047] Comparative Example 1, Preparation of Jerusalem artichoke polysaccharide: Step a, adding 10 L of water to 1 kg of Jerusalem artichoke, adjusting the pH to 5.0, refluxing and extracting twice at 70° C., filtering, and combining the two filtrates to obtain an extract; Step b, adjusting the pH of the extract obtained in step a to 7.0, extracting with ethyl acetate, taking the lower layer of solution, removing the residual ethyl acetate, concentrating at 70° C., and using the concentrate for later use; Step c, loading the concentrated solution obtained in step b onto a silica gel column, using a mixed solution of n-butanol: water: formic acid = 0.23:10:0.07 by volume as an elution solvent, collecting the eluate, removing n-butanol from the eluate and concentrating it, adjusting the pH to 6.5, adding 0.33 g / L of glucose oxidase (enzyme activity of 2000 u / g) and 1.3 g / L of β-glucanase (enzyme activity of 20000 u / g), performing enzymolysis at 40° C. for 5 h, adding 7 g / L of oryzanol and heating to 80° C. to inactivate the enzyme, adjusting the alcohol content of the enzymolysis solution to 20%, 30%, 50%, and 70% with anhydrous ethanol in turn, standing for 5 h after each adjustment of the alcohol content, centrifuging, and taking the precipitate to obtain Jerusalem artichoke polysaccharide.

[0048] Prepare the composition and essence containing complex peptides according to the following ratio and method (1 kg)

[0049] Step 1: Add palmitoyl tripeptide-8, palmitoyl tetrapeptide-7, vitamin E, and PEG hydrogenated castor oil into a container, heat to 60°C and stir until dissolved to prepare the oil phase.

[0050] Step 2: Add tripeptide-1, acetyl dipeptide-1, glycerol and water into a container and stir at room temperature to dissolve to prepare an aqueous phase.

[0051] Step 3: Add the water phase to the oil phase, stir evenly, add polysaccharides and phospholipids, and use acetic acid and triethanolamine to adjust the pH value to 5.0.

[0052] Step 4: Filter (0.22μm filter membrane) and package in a GMP workshop or clean bench to obtain the essence.

[0053] Comparative Example 2, Preparation of Jerusalem artichoke polysaccharide: Step a, adding 10 L of water to 1 kg of Jerusalem artichoke, adjusting the pH to 5.0, refluxing and extracting twice at 60-80° C., filtering, and combining the two filtrates to obtain an extract; Step b, adjusting the pH of the extract obtained in step a to 7.0, extracting with ethyl acetate, taking the lower layer of solution, removing the residual ethyl acetate, concentrating at 70° C., and using the concentrate for later use; Step c, loading the concentrated solution obtained in step b onto a silica gel column, using a mixed solution of n-butanol: water: formic acid = 0.23:10:0.07 by volume as an elution solvent, collecting the eluate, removing n-butanol from the eluate and concentrating it, adjusting the pH to 6.5, adding 0.33 g / L of glucose oxidase (enzyme activity of 2000 u / g) and 1.3 g / L of β-glucanase (enzyme activity of 20000 u / g), performing enzymolysis at 40° C. for 5 h, adding 7 g / L of oryzanol and heating to 80° C. to inactivate the enzyme, adjusting the alcohol content of the enzymolysis solution to 20%, 30%, 50%, and 70% with anhydrous ethanol in turn, standing for 5 h after each adjustment of the alcohol content, centrifuging, and taking the precipitate to obtain Jerusalem artichoke polysaccharide.

[0054] Prepare the composition and essence containing complex peptides according to the following ratio and method (1 kg)

[0055] Step 1: Add palmitoyl tripeptide-8, palmitoyl tetrapeptide-7, vitamin E, and PEG hydrogenated castor oil into a container, heat to 60°C and stir until dissolved to prepare the oil phase.

[0056] Step 2: Add tripeptide-1, acetyl dipeptide-1, glycerol and water into a container and stir at room temperature to dissolve to prepare an aqueous phase.

[0057] Step 3: Add the water phase to the oil phase, stir evenly, add polysaccharides and phospholipids, and use acetic acid and triethanolamine to adjust the pH value to 5.0.

[0058] Step 4: Filter (0.22μm filter membrane) and package in a GMP workshop or clean bench to obtain the essence.

[0059] Comparative Example 3, Preparation of Jerusalem artichoke polysaccharide: Step a, adding 10 L of water to 1 kg of Jerusalem artichoke, adjusting the pH to 5.0, refluxing and extracting twice at 60-80° C., filtering, and combining the two filtrates to obtain an extract; Step b, adjusting the pH of the extract obtained in step a to 7.0, extracting with ethyl acetate, taking the lower layer of solution, removing the residual ethyl acetate, concentrating at 70° C., and using the concentrate for later use; Step c, loading the concentrated solution obtained in step b onto a silica gel column, using a mixed solution of n-butanol: water: formic acid = 0.23:10:0.07 by volume as an elution solvent, collecting the eluate, removing n-butanol from the eluate and concentrating it, adjusting the pH to 6.5, adding 0.33 g / L of glucose oxidase (enzyme activity of 2000 u / g) and 1.3 g / L of β-glucanase (enzyme activity of 20000 u / g), performing enzymolysis at 40° C. for 5 h, adding 7 g / L of oryzanol and heating to 80° C. to inactivate the enzyme, adjusting the alcohol content of the enzymolysis solution to 20%, 30%, 50%, and 70% with anhydrous ethanol in turn, standing for 5 h after each adjustment of the alcohol content, centrifuging, and taking the precipitate to obtain Jerusalem artichoke polysaccharide.

[0060] Prepare the composition and essence containing complex peptides according to the following ratio and method (1 kg)

[0061] Step 1: Add palmitoyl tripeptide-8, palmitoyl tetrapeptide-7, vitamin E, and PEG hydrogenated castor oil into a container, heat to 60°C and stir until dissolved to prepare the oil phase.

[0062] Step 2: Add tripeptide-1, acetyl dipeptide-1, glycerol and water into a container and stir at room temperature to dissolve to prepare an aqueous phase.

[0063] Step 3: Add the water phase to the oil phase, stir evenly, add polysaccharides and phospholipids, and use acetic acid and triethanolamine to adjust the pH value to 5.0.

[0064] Step 4: Filter (0.22μm filter membrane) and package in a GMP workshop or clean bench to obtain the essence.

[0065] Comparative Example 4, Preparation of Jerusalem artichoke polysaccharide: Step a, adding 10 L of water to 1 kg of Jerusalem artichoke, adjusting the pH to 5.0, refluxing and extracting twice at 60-80° C., filtering, and combining the two filtrates to obtain an extract; Step b, adjusting the pH of the extract obtained in step a to 7.0, extracting with ethyl acetate, taking the lower layer of solution, removing the residual ethyl acetate, concentrating at 70° C., and using the concentrate for later use; Step c, loading the concentrated solution obtained in step b onto a silica gel column, using a mixed solution of n-butanol: water: formic acid = 0.23:10:0.07 by volume as an elution solvent, collecting the eluate, removing n-butanol from the eluate and concentrating it, adjusting the pH to 6.5, adding 0.33 g / L of glucose oxidase (enzyme activity of 2000 u / g) and 1.3 g / L of β-glucanase (enzyme activity of 20000 u / g), performing enzymolysis at 40° C. for 5 h, adding 7 g / L of oryzanol and heating to 80° C. to inactivate the enzyme, adjusting the alcohol content of the enzymolysis solution to 20%, 30%, 50%, and 70% with anhydrous ethanol in turn, standing for 5 h after each adjustment of the alcohol content, centrifuging, and taking the precipitate to obtain Jerusalem artichoke polysaccharide.

[0066] Prepare the composition and essence containing complex peptides according to the following ratio and method (1 kg)

[0067] Step 1: Add palmitoyl tripeptide-8, palmitoyl tetrapeptide-7, vitamin E, and PEG hydrogenated castor oil into a container, heat to 60°C and stir until dissolved to prepare the oil phase.

[0068] Step 2: Add tripeptide-1, acetyl dipeptide-1, glycerol and water into a container and stir at room temperature to dissolve to prepare an aqueous phase.

[0069] Step 3: Add the water phase to the oil phase, stir evenly, add polysaccharides and phospholipids, and use acetic acid and triethanolamine to adjust the pH value to 5.0.

[0070] Step 4: Filter (0.22μm filter membrane) and package in a GMP workshop or clean bench to obtain the essence.

[0071] Comparative Example 5: A composition containing a complex peptide and an essence (1 kg) were prepared according to the following ratio and method:

[0072] Step 1: Add palmitoyl tripeptide-8, palmitoyl tetrapeptide-7, vitamin E, and PEG hydrogenated castor oil into a container, heat to 60°C and stir until dissolved to prepare the oil phase.

[0073] Step 2: Add tripeptide-1, acetyl dipeptide-1, glycerol and water into a container and stir at room temperature to dissolve to prepare an aqueous phase.

[0074] Step 3: Add the water phase to the oil phase, stir evenly, add phospholipids, and adjust the pH value to 5.0 using acetic acid and triethanolamine.

[0075] Step 4: Filter (0.22μm filter membrane) and package in a GMP workshop or clean bench to obtain the essence.

[0076] Comparative Example 6, Preparation of Jerusalem artichoke polysaccharide: Step a, adding 10 L of water to 1 kg of Jerusalem artichoke, reflux extraction twice at 70° C., filtering, and combining the two filtrates to obtain an extract; Step b, adjusting the pH of the extract obtained in step a to 7.0, extracting with ethyl acetate, taking the lower layer of solution, removing the residual ethyl acetate, concentrating at 70° C., and using the concentrate for later use; Step c, loading the concentrated solution obtained in step b onto a silica gel column, using a mixed solution of n-butanol: water: formic acid = 0.23:10:0.07 by volume as an elution solvent, collecting the eluate, removing n-butanol from the eluate and concentrating it, adjusting the alcohol content to 50%, standing for 5 hours, centrifuging, and taking the precipitate to obtain Jerusalem artichoke polysaccharide.

[0077] Prepare the composition and essence containing complex peptides according to the following ratio and method (1 kg)

[0078] Step 1: Add palmitoyl tripeptide-8, palmitoyl tetrapeptide-7, vitamin E, and PEG hydrogenated castor oil into a container, heat to 60°C and stir until dissolved to prepare the oil phase.

[0079] Step 2: Add tripeptide-1, acetyl dipeptide-1, glycerol and water into a container and stir at room temperature to dissolve to prepare an aqueous phase.

[0080] Step 3: Add the water phase to the oil phase, stir evenly, add Jerusalem artichoke polysaccharide and phospholipids, and use acetic acid and triethanolamine to adjust the pH value to 5.0.

[0081] Step 4: Filter (0.22μm filter membrane) and package in a GMP workshop or clean bench to obtain the essence.

[0082] Comparative Example 7, Preparation of Jerusalem artichoke polysaccharide: Add 10L of water to 1kg of Jerusalem artichoke, reflux extract twice at 70℃, filter, and combine the two filtrates to obtain an extract; adjust the pH to 6.5, add 0.33g / L of glucose oxidase (enzyme activity is 2000u / g) and 1.3g / L of β-glucanase (enzyme activity is 20000u / g), and enzymolyze at 40℃ for 5h. The alcohol content of the enzymolyzate is adjusted to 20%, 30%, 50%, and 70% with anhydrous ethanol in turn. After each adjustment of the alcohol content, let it stand for 5h, centrifuge, and take the precipitate to obtain Jerusalem artichoke polysaccharide.

[0083] Prepare the composition and essence containing complex peptides according to the following ratio and method (1 kg)

[0084] Step 1: Add palmitoyl tripeptide-8, palmitoyl tetrapeptide-7, vitamin E, and PEG hydrogenated castor oil into a container, heat to 60°C and stir until dissolved to prepare the oil phase.

[0085] Step 2: Add tripeptide-1, acetyl dipeptide-1, glycerol and water into a container and stir at room temperature to dissolve to prepare an aqueous phase.

[0086] Step 3: Add the water phase to the oil phase, stir evenly, add Jerusalem artichoke polysaccharide and phospholipids, and use acetic acid and triethanolamine to adjust the pH value to 5.0.

[0087] Step 4: Filter (0.22μm filter membrane) and package in a GMP workshop or clean bench to obtain the essence.

[0088] Comparative Example 8, Preparation of Jerusalem artichoke polysaccharide: Step a, adding 10 L of water to 1 kg of Jerusalem artichoke, adjusting the pH to 5.0, refluxing and extracting twice at 70° C., filtering, and combining the two filtrates to obtain an extract; Step b, adjusting the pH of the extract obtained in step a to 7.0, extracting with ethyl acetate, taking the lower layer of solution, removing the residual ethyl acetate, concentrating at 70° C., and using the concentrate for later use; Step c, loading the concentrated solution obtained in step b onto a silica gel column, using a mixed solution with a volume ratio of n-butanol: water: formic acid = 0.23:10:0.07 as an elution solvent, collecting the eluate, removing n-butanol from the eluate and concentrating it, adjusting the pH to 6.5, adding 0.33 g / L of glucose oxidase (enzyme activity of 2000 u / g) and 1.3 g / L of β-glucanase (enzyme activity of 20000 u / g), performing enzymolysis at 40° C. for 5 h, heating to 80° C. to inactivate the enzyme, adjusting the alcohol content of the enzymolysis solution to 20%, 30%, 50%, and 70% with anhydrous ethanol in turn, standing for 5 h after each adjustment of the alcohol content, centrifuging, and taking the precipitate to obtain Jerusalem artichoke polysaccharide.

[0089] Prepare the composition and essence containing complex peptides according to the following ratio and method (1 kg)

[0090] Step 1: Add palmitoyl tripeptide-8, palmitoyl tetrapeptide-7, and PEG hydrogenated castor oil into a container, heat to 60°C and stir until dissolved to prepare the oil phase.

[0091] Step 2: Add tripeptide-1, acetyl dipeptide-1, glycerol and water into a container and stir at room temperature to dissolve to prepare an aqueous phase.

[0092] Step 3: Add the water phase to the oil phase, stir evenly, add Jerusalem artichoke polysaccharide and phospholipids, and use acetic acid and triethanolamine to adjust the pH value to 5.0.

[0093] Step 4: Filter (0.22μm filter membrane) and package in a GMP workshop or clean bench to obtain the essence.

[0094] Performance Testing 1. The essence prepared by the present invention is used to repair the face of hormone-induced face In this experiment, 80 volunteers aged between 25 and 30 were selected. Due to the use of cosmetics containing hormones, their faces showed obvious redness, swelling, inflammation, itching and other phenomena to varying degrees.

[0095] The volunteers were randomly divided into 8 groups, with 10 people in each group. After cleansing their faces in the morning and evening every day, they used the essences prepared by Examples 4 to 6, Comparative Examples 1 to 4, and a repairing anti-allergic essence of a well-known brand on the market for 28 consecutive days to test the repair effect of each essence.

[0096] The American Visia skin tester was used to test and record the skin redness of the subjects.

[0097] Table 1 Improvement of hormone-induced facial by different essences

[0098] It can be seen from the results in Table 1 that the four polypeptides of different action mechanism categories in the present invention have obvious synergistic effects, and the efficacy of test 1 to test 3 groups in repairing hormone face is significantly higher than that of test 4 to test 7 groups and a well-known brand of soothing repair essence on the market.

[0099] 2. Anti-wrinkle effect of the essence prepared by the present invention In this experiment, 80 volunteers aged 40-45 years old were selected and randomly divided into 8 groups, with 10 people in each group. After cleansing their faces in the morning and evening every day, the essences prepared in Examples 4 to 6, Comparative Examples 1 to Comparative Examples 4 and an anti-wrinkle essence of a well-known commercial brand were used for 28 consecutive days to test the anti-wrinkle effect of each essence.

[0100] The American Visia skin tester was used to test and record the skin red area of ​​the subjects.

[0101] Table 2 Comparison of anti-wrinkle effects of different essences

[0102] From the results in Table 2, it can be seen that the combination of the four polypeptides in the present invention has a better effect. The anti-wrinkle effect of groups 1 to 3 on the face is significantly higher than that of groups 4 to 7 and a well-known brand of anti-wrinkle essence on the market, indicating that the four polypeptides of the present invention act synergistically to achieve a higher anti-wrinkle effect.

[0103] 3. Antioxidant stability of different essences The composite peptide obtained in Example 1 and the essences obtained in Example 4 and Comparative Examples 5 to 8 were exposed to natural light at room temperature and ultraviolet light at room temperature, respectively, and the effects of the composite peptide and the essence on ·OH and ·O at different time points were measured. 2- clearance rate.

[0104] Results Figure 1-Figure 4 It can be seen that the composite peptide in Example 1 and the essences of Comparative Examples 5 to 8 will have a delayed effect on ·OH and ·O under natural light and ultraviolet light at room temperature over time. 2- The clearance rate of ·OH and ·O gradually decreased, and its physiological activity gradually decreased; while the essence in Example 4 2- Although the clearance rate decreased, the decrease was small and the physiological activity did not change much. It can be seen that the addition of Jerusalem artichoke polysaccharide and vitamin E in the present invention helps to improve the physiological activity and stability of the composite peptide.

Claims

1. A composite peptide with anti-inflammatory and soothing effects, characterized in that: It contains the following ingredients: tripeptide-1, acetyl dipeptide-1, palmitoyl tetrapeptide-7, and palmitoyl tripeptide-8; the mass ratio of the tripeptide-1, acetyl dipeptide-1, palmitoyl tetrapeptide-7, and palmitoyl tripeptide-8 is (0.5-1.5):(0.6-1.5):(1-2):(0.5-1.5).

2. Use of the composite peptide according to claim 1 in the preparation of cosmetics with anti-inflammatory and soothing effects.

3. A composition containing the composite peptide according to claim 1, characterized in that: In terms of mass fraction, it mainly contains the following ingredients: 0.8-1.2% tripeptide-1, 0.8-1.5% acetyl dipeptide-1, 1.0-1.5% palmitoyl tetrapeptide-7, 0.9-1.3% palmitoyl tripeptide-8, 3-6% glycerin, 3-8% Jerusalem artichoke polysaccharide, 0.5-1.5% PEG hydrogenated castor oil, 0.5-1.0% phospholipids, 0.1-0.14% vitamin E, and the balance is purified water.

4. The composition according to claim 3, characterized in that The preparation method of the Jerusalem artichoke polysaccharide is: Step a, adding water to Jerusalem artichoke, adjusting the pH to 4.5-5.0, and refluxing and extracting at 60-80° C. to obtain an extract; Step b, adjusting the pH of the extract obtained in step a to 7.0-7.5, extracting with ethyl acetate, taking the lower layer of solution, removing the residual ethyl acetate, concentrating at 60-80° C., and using the concentrate for later use; Step c, separating the concentrated solution obtained in step b by using a silica gel column, using a mixed solution of n-butanol-water-formic acid as an elution solvent, collecting the eluate, removing n-butanol from the eluate and concentrating it, adjusting the pH to 6.5-7.0, adding glucose oxidase and β-glucanase, performing enzymolysis at 35-40° C. for 4-6 hours, adding oryzanol and heating to 80-90° C. to inactivate the enzyme, adjusting the alcohol content of the enzymolysis solution to 20%, 30%, 50%, and 70% by using anhydrous ethanol in sequence, standing it for 4-6 hours after each adjustment of the alcohol content, centrifuging it, and taking the precipitate to obtain Jerusalem artichoke polysaccharide.

5. The composition according to claim 4, characterized in that In the step c of the method for preparing Jerusalem artichoke polysaccharide, the volume ratio of n-butanol, water and formic acid in the mixed solution of n-butanol-water-formic acid is (0.2-0.25):10:(0.06-0.08).

6. The composition according to claim 4, characterized in that In step c of the Jerusalem artichoke polysaccharide preparation method, the amount of glucose oxidase added is 0.3-0.35 g / L, and the amount of β-glucanase added is 1-1.5 g / L.

7. The composition according to claim 4, characterized in that In step c of the Jerusalem artichoke polysaccharide preparation method, the enzyme activity of glucose oxidase is 2000u / g, and the enzyme activity of β-glucanase is 20000u / g.

8. The composition according to claim 4, characterized in that In step c of the Jerusalem artichoke polysaccharide preparation method, the amount of oryzanol added is 5-8 g / L.

9. A method for preparing the composition according to any one of claims 3 to 8, characterized in that: The following steps are involved: Step 1: Add palmitoyl tripeptide-8, palmitoyl tetrapeptide-7, vitamin E, and PEG hydrogenated castor oil into a container and heat to 60°C and stir until dissolved to prepare the oil phase; Step 2: Add tripeptide-1, acetyl dipeptide-1, glycerol and water into a container and stir at room temperature to dissolve to prepare an aqueous phase; Step 3: Add the aqueous phase obtained in step 2 to the oil phase obtained in step 1, stir evenly, add Jerusalem artichoke polysaccharide and phospholipids, and adjust the pH value to 5.0 to obtain a composition.

10. Use of the composition according to any one of claims 3 to 8 in the preparation of cosmetics with anti-inflammatory and soothing effects.

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