Cell active peptide composition with anti-allergy and redness-reducing effects, its preparation method and medical aesthetic application

Abstract

The present invention provides a cell active peptide composition with anti-allergic and redness-reducing effects, its preparation method and medical aesthetic applications, belonging to the fields of medicine and beauty technology. The composition includes: dipeptide-1, tripeptide-1, palmitoyl tripeptide-8, palmitoyl tetrapeptide-7, sh-pentapeptide-1, hydrolyzed silk, mussel extract, dipeptide-2, acetyl tetrapeptide-3, ceramide, 4-tert-butylcyclohexanol, bisabolol, sodium hyaluronate, etc. The composition can repair skin barrier damage; improve skin nerve dysfunction; strengthen immune function and eliminate skin inflammation; enhance the elasticity of the capillary wall and repair telangiectasia. The composition of the present invention is applied to dermatology and skin beauty, can effectively reduce the average value of skin flushing, the area of red area and the redness a value, strengthen the brick-wall structure, has a mild effect, and has the functions of anti-allergic redness reduction, soothing and calming, and immune regulation.

Description

Technical Field

[0001] The present invention belongs to the field of medicine and cosmetology technology, and specifically relates to a cell active peptide composition with anti-allergic and redness-removing effects, a preparation method thereof, and medical and cosmetology applications. Background Art

[0002] Skin, as the largest organ in the human body, plays multiple roles: it is our defense shield, protecting us from external aggression; it is a transmitter of sensation, allowing us to experience subtle changes in touch; it is an excretion channel, helping us to eliminate waste from the body; it is part of the immune system, resisting the invasion of pathogens. However, the skin is also extremely vulnerable to physical factors such as high temperature, strong ultraviolet radiation, high altitude climate, and chemical factors such as steroid hormones, preservatives, surfactants, etc., all of which may cause skin inflammation.

[0003] A healthy skin barrier has excellent tolerance to physical and chemical stimuli, and has good self-repair and regulation capabilities. In contrast, sensitive skin reacts more violently to minor external stimuli, and may experience itching, tingling, pins and needles, burning, tightness, dry skin, erythema, fine scales, and easy flushing. The skin is not only a physical barrier for the human body, but also an active neuro-endocrine-immune organ that involves the interaction of the nervous, endocrine, and immune systems. The causes and mechanisms of sensitive skin include damage to the skin barrier, abnormalities in the nervous sensory system, increased vascular reactivity, and immune inflammatory responses. Existing technologies lack comprehensive care for sensitive skin from the aspects of nervous system recovery, reduced vascular permeability, skin barrier repair, and inflammation control.

[0004] Therefore, there is an urgent need for a cell active peptide composition with anti-allergic and redness-removing effects, as well as a preparation method and medical and aesthetic applications thereof. Summary of the invention

[0005] The purpose of the present invention is to provide a cell active peptide composition with anti-allergic and redness-removing effects, a preparation method thereof, and medical and aesthetic applications.

[0006] In order to achieve the above object, the present invention provides the following technical solutions:

[0007] A cell-active peptide composition with anti-allergy and redness-reducing effects, comprising the following components in parts by weight: 1-2 parts of tripeptide-1, 0.1-0.8 part of ceramide, 2-3 parts of hydrolyzed silk, 1.3-2.6 parts of mussel extract, 0.0001-0.2 part of sodium hyaluronate, 0.0001-1 part of 4-tert-butylcyclohexanol, 0.0001-0.2 part of dipeptide-1, 0.0001-0.2 part of palmitoyl tripeptide-8, 0.0001-0.2 part of palmitoyl tetrapeptide-7, 0.0001-0.15 part of sh-pentapeptide-1, 0.0001-0.15 part of dipeptide-2, 0.0001-0.2 part of bisabolol, 0.0001-0.15 part of acetyl tetrapeptide-3;

[0008] The preparation method of the mussel extract is as follows:

[0009] (1) Take out the fresh mussel meat from the green-lipped mussel, break up the mussel meat, and add water 5-6 times the weight of the mussel meat to obtain a mixture;

[0010] (2) Add a complex enzyme to the mixture for enzymatic hydrolysis, hydrolyze at 30-35 °C for 1-2 days, inactivate the enzyme, and obtain an enzymatic hydrolysis product;

[0011] (3) Add a complex bacterium to the enzymatic hydrolysis product for fermentation, ferment at 38-40 °C for 3-5 days, sterilize, and obtain a fermentation product;

[0012] (4) First filter the fermentation product through a 0.45 μm filter membrane to obtain a filtrate, then ultrafilter the filtrate through a 10 kDa ultrafiltration membrane to obtain an ultrafiltrate, and perform spray drying to obtain the mussel extract.

[0013] Furthermore, the complex enzyme includes pectinase, bromelain, and chitosanase.

[0014] Furthermore, the enzyme activity of pectinase in the mixture is 70 U / mL - 90 U / mL; the enzyme activity of bromelain in the mixture is 150 U / mL - 170 U / mL; the enzyme activity of chitosanase in the mixture is 110 U / mL - 130 U / mL.

[0015] Furthermore, the complex bacterium includes Bifidobacterium longum, Bifidobacterium lactis, and Lactobacillus rhamnosus.

[0016] Furthermore, the viable count of Bifidobacterium longum in the enzymatic hydrolysis product is 3.0×10 9 CFU / mL - 5.0×10 9 CFU / mL, the viable count of Bifidobacterium lactis is 6.0×10 8 CFU / mL - 8.0×10 8 CFU / mL, and the viable count of Lactobacillus rhamnosus is 0.5×10 9CFU / mL - 1.5×10 9 CFU / mL.

[0017] Research shows that mussel extract has potential anti - allergic effects. However, mussel extracts on the market are usually extracted using organic reagents, which have problems such as low extraction efficiency and damage to thermosensitive components. When added to the system of the present invention, the anti - allergic effect is not ideal. In the present invention, mussel meat is first enzymolyzed using a composite enzyme and then fermented using a composite bacterium, and the prepared mussel extract has a better anti - allergic effect. Enzymolyzing mussel meat with a composite enzyme first in the present invention can effectively decompose proteins and other macromolecules in mussel meat, making it easier to be absorbed and possibly reducing the macromolecular structures that cause allergic reactions. Subsequently, fermentation with a composite bacterium is carried out. The fermentation process can not only further degrade proteins but also produce some beneficial metabolites, such as short - chain fatty acids, etc. These metabolites may help enhance the skin barrier function and reduce allergic reactions. The present invention improves the bioavailability of the extract and at the same time changes the components that may cause allergic reactions, thereby reducing the risk of allergy, which is of great significance to consumers who hope to obtain skin care benefits from marine organisms but are worried about allergic reactions. However, the improvement of the system's redness - reducing effect by the mussel extract prepared in the present invention in combination with other components is not as good as adding commercially available mussel extract.

[0018] Furthermore, the preparation method of hydrolyzed silk is as follows:

[0019] S1. Clean and boil the mulberry silkworm cocoon shell to degum it to obtain silk fibroin.

[0020] S2. Divide the silk fibroin into three groups, namely silk fibroin group A, silk fibroin group B, and silk fibroin group C. Add 7 - 10 parts by weight of water and Bifidobacterium breve with a viable count of 10 9 CFU / mL - 10 10 CFU / mL to 1 part by weight of silk fibroin group A, and ferment at 40 - 45°C for 40 - 50 h, then sterilize to obtain the fermentation broth of silk fibroin group A; add 7 - 10 parts by weight of water and Lactobacillus casei with a viable count of 10 9 CFU / mL - 10 10 CFU / mL to 1 part by weight of silk fibroin group B, and ferment at 40 - 45°C for 60 - 70 h, then sterilize to obtain the fermentation broth of silk fibroin group B; add 7 - 10 parts by weight of water and Lactobacillus brevis with a viable count of 10 9 CFU / mL - 10 10 CFU / mL to 1 part by weight of silk fibroin group C, and ferment at 40 - 45°C for 20 - 30 h, then sterilize to obtain the fermentation broth of silk fibroin group C.

[0021] S3. Respectively decolorize the fibroin A group fermentation broth, fibroin B group fermentation broth, and fibroin C group fermentation broth with activated carbon, and filter them through a 0.22 μm microfiltration membrane to obtain the decolorized fibroin A group solution, decolorized fibroin B group solution, and decolorized fibroin C group solution respectively;

[0022] S4. Respectively concentrate and dry the decolorized fibroin A group solution, decolorized fibroin B group solution, and decolorized fibroin C group solution at low temperature, and then perform ultraviolet sterilization treatment to obtain the fibroin A group powder, fibroin B group powder, and fibroin C group powder;

[0023] S5. Thoroughly mix the fibroin A group powder, fibroin B group powder, and fibroin C group powder in a specific weight ratio to obtain hydrolyzed silk.

[0024] Furthermore, the weight ratio of the fibroin A group powder, fibroin B group powder, and fibroin C group powder in step S5 is 1:(1.2 - 1.4):(0.5 - 0.8).

[0025] In order to improve the redness-reducing effect of the products of the present invention, the present invention attempts to add hydrolyzed silk to assist the mussel extract to jointly improve redness reduction, but the effect is not ideal. By fermenting fibroin raw materials with different bacteria, hydrolyzed silk compositions with different compositions can be obtained. The hydrolyzed silk is obtained by compounding and added to the system of the present invention, and it can cooperate with the other components to improve the redness-reducing effect of the products. The hydrolyzed silk prepared by the present invention contains more active ingredients that are helpful for skin recovery and soothing, and can produce a synergistic effect with other components such as the mussel extract, and can more effectively relieve skin redness and reduce inflammatory reactions.

[0026] Furthermore, the weight ratio of the mussel extract, hydrolyzed silk, and tripeptide-1 is (1.2 - 1.5):(2.4 - 2.8):(1.9 - 2.2).

[0027] It was found in the experiments that the addition amounts of the mussel extract and hydrolyzed silk would affect the storage stability of the products, and unpleasant odors or color darkening would easily occur if the products were not stored properly. Through a large number of experiments, it was found that when the mussel extract, hydrolyzed silk, and tripeptide-1 were compounded in a specific ratio, the stability of the products could be improved. The adhesiveness and antioxidant properties of the mussel extract of the present invention are combined with the moisturizing property and film-forming property of hydrolyzed silk to form a more stable system. Tripeptide-1 is generally relatively stable and not easily decomposed, and can cooperate with other components to improve the stability of the overall formulation.

[0028] Tripeptide-1: Helps repair damaged skin barriers by promoting the synthesis of the extracellular matrix and enhancing the skin's regenerative ability.

[0029] Ceramide: Ceramide is an important component of the skin barrier. Supplementing ceramide can help repair and strengthen the skin barrier.

[0030] Sodium hyaluronate: It has a strong hydration ability. Topical HA can regulate the differentiation of keratinocytes, inhibit the inflammatory level, and enhance the production of endogenous HA, thereby repairing the skin barrier.

[0031] 4-tert-Butylcyclohexanol: 4-tert-Butylcyclohexanol can immediately relieve skin pain and burning sensation by inhibiting TRPV-1. Research shows that 4-tert-Butylcyclohexanol significantly reduces the activation of TRPV-1 in vitro and has a significant immediate anti-tingling / anti-burning effect in vivo.

[0032] Dipeptide-1: It has analgesic, neuroprotective and neuromodulatory properties. It is synthesized at the nerve endings and acts by stimulating the release of methionine enkephalin from nerve cells. Methionine enkephalin is an opioid (i.e., relaxation, pain relief) messenger molecule with neuro- and immunomodulatory functions.

[0033] Palmitoyl tripeptide-8: Derived from pro-opiomelanocortin (POMC), it can soothe and relieve irritated skin, restoring the skin to its normal sensitivity threshold.

[0034] Palmitoyl tetrapeptide-7: Reduces the secretion of IL-6, reduces inflammation after UVB exposure, and stimulates the production of laminin IV and V and collagen VII.

[0035] SH-Pentapeptide-1: Also known as thymopentin (TP5), with the amino acid sequence RKDVY, it is the amino acid fragment at positions 32-36 of thymopoietin II (TPII). It has similar biological activities to TPII and can bidirectionally regulate the imbalanced immune system. It is an effective immunomodulator.

[0036] Dipeptide-2: An effective ACE inhibitor, it promotes blood circulation by reducing blood vessel constriction.

[0037] Acetyl tetrapeptide-3: Promotes the generation of vascular endothelium and increases the elasticity of capillaries.

[0038] Bisabolol: Bisabolol is a natural monoterpene alcohol extracted from Chamomile, widely used in cosmetics and pharmaceuticals, especially for relieving sensitive skin, reducing skin inflammation and promoting skin health.

[0039] Furthermore, the cell active peptide composition includes the following components in parts by weight: 1 - 2 parts of Tripeptide - 1, 0.1 - 0.8 part of ceramide, 2 - 3 parts of hydrolyzed silk, 1.3 - 2.6 parts of mussel extract, 0.0001 - 0.2 part of sodium hyaluronate, 0.0001 - 1 part of 4 - tert - butylcyclohexanol, 0.0001 - 0.2 part of Dipeptide - 1, 0.0001 - 0.2 part of Palmitoyl Tripeptide - 8, 0.0001 - 0.2 part of Palmitoyl Tetrapeptide - 7, 0.0001 - 0.15 part of sh - Pentapeptide - 1, 0.0001 - 0.15 part of Dipeptide - 2, 0.0001 - 0.2 part of bisabolol, 0.0001 - 0.15 part of Acetyl Tetrapeptide - 3.

[0040] The present invention also provides a preparation method of the cell active peptide composition. The preparation method includes the following steps: weighing the corresponding components according to the weight ratio, mixing them evenly, and sterilizing to obtain the cell active peptide composition.

[0041] The present invention also provides the application of the cell active peptide composition in skin care products.

[0042] The present invention also provides the application of the cell active peptide composition in medical beauty.

[0043] The present invention also provides the application of the cell active peptide composition in the preparation of cosmetics.

[0044] The present invention also provides the application of the cell active peptide composition in the preparation of topical skin drugs.

[0045] The present invention also provides the application of the cell active peptide composition in the preparation of beauty products.

[0046] Compared with the prior art, the advantages and beneficial effects of the present invention are as follows:

[0047] 1. Mussel extracts on the market are usually extracted using organic reagents, which have problems such as low extraction efficiency and damage to thermosensitive components. When added to the system of the present invention, the anti - allergy effect is not ideal. In the present invention, mussel meat is first enzymolyzed using a composite enzyme and then fermented using a composite bacterium, and the prepared mussel extract has a better anti - allergy effect.

[0048] 2. By using different bacteria to ferment silk fibroin raw materials, the present invention can obtain hydrolyzed silk compositions with different compositions. Through compounding to obtain hydrolyzed silk and adding it to the system of the present invention, through synergistic action with other components, the redness - reducing effect of the product can be improved.

[0049] 3. By compounding mussel extract, hydrolyzed silk and Tripeptide - 1 in a specific ratio, the present invention can improve the stability of the product. Detailed implementation manners

[0050] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Apparently, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0051] The enzymes and bacteria used in the present invention are commercially available products, specifically as follows.

[0052] Bifidobacterium longum, preservation number: SHBCCD24310, purchased from Shanghai Culture Collection of Microorganisms.

[0053] Bifidobacterium lactis, preservation number: SHBCCD24728, purchased from Shanghai Culture Collection of Microorganisms.

[0054] Lactobacillus rhamnosus, preservation number: SHBCCD73158, purchased from Shanghai Culture Collection of Microorganisms.

[0055] Bifidobacterium breve, preservation number: SHBCCD24424JCM7017, purchased from Shanghai Culture Collection of Microorganisms.

[0056] Lactobacillus casei, preservation number: SHBCCD24737, purchased from Shanghai Culture Collection of Microorganisms.

[0057] Lactobacillus brevis, preservation number: SHBCCD14346, purchased from Shanghai Culture Collection of Microorganisms.

[0058] All the enzymes used in the present invention are purchased from Shanghai Yuanye Bio-Technology Co., Ltd.

[0059] Example 1

[0060] This example provides a cell active peptide composition, including the following components in parts by weight: 1.3 parts of tripeptide-1, 0.5 part of ceramide, 2.6 parts of hydrolyzed silk, 2.0 parts of mussel extract, 0.003 part of sodium hyaluronate, 0.1 part of 4-tert-butylcyclohexanol, 0.0009 part of dipeptide-1, 0.05 part of palmitoyl tripeptide-8, 0.008 part of palmitoyl tetrapeptide-7, 0.12 part of sh-pentapeptide-1, 0.09 part of dipeptide-2, 0.02 part of bisabolol, 0.03 part of acetyl tetrapeptide-3.

[0061] The preparation method of the mussel extract is as follows:

[0062] (1) Take out the fresh mussel meat from the green-lipped mussel, break the mussel meat, and add 6 times the weight of water of the mussel meat to obtain a mixture;

[0063] (2) Add a complex enzyme to the mixture for enzymatic hydrolysis, hydrolyze at 32 °C for 2 days, and after inactivating the enzyme, obtain an enzymatic hydrolysis product;

[0064] (3) Add compound bacteria to the enzymatic hydrolysate and ferment at 39 °C for 4 days, then sterilize to obtain the fermentation product;

[0065] (4) First filter the fermentation product through a 0.45 μm filter membrane to obtain a filtrate, then ultrafilter the filtrate through an ultrafiltration membrane with a molecular weight cut-off of 10 kDa to obtain an ultrafiltrate, and perform spray drying to obtain the mussel extract.

[0066] The compound enzyme includes pectinase, bromelain, and chitosanase.

[0067] The enzyme activity of pectinase in the mixture is 80 U / mL; the enzyme activity of bromelain in the mixture is 160 U / mL; the enzyme activity of chitosanase in the mixture is 120 U / mL.

[0068] The compound bacteria include Bifidobacterium longum, Bifidobacterium lactis, and Lactobacillus rhamnosus.

[0069] The viable count of Bifidobacterium longum added to the enzymatic hydrolysate is 4.0×10 9 CFU / mL, the viable count of Bifidobacterium lactis is 7.0×10 8 CFU / mL, and the viable count of Lactobacillus rhamnosus is 1×10 9 CFU / mL.

[0070] The preparation method of hydrolyzed silk is as follows:

[0071] S1. Wash and boil the mulberry silk cocoon shell to degum it, and obtain fibroin.

[0072] S2. Divide the fibroin into three groups, namely fibroin group A, fibroin group B, and fibroin group C; add 8 parts by weight of water and Bifidobacterium breve with a viable count of 10 10 CFU / mL to 1 part by weight of fibroin group A, ferment at 42 °C for 45 h, and sterilize to obtain the fermentation broth of fibroin group A; add 9 parts by weight of water and Lactobacillus casei with a viable count of 10 9 CFU / mL to 1 part by weight of fibroin group B, ferment at 42 °C for 65 h, and sterilize to obtain the fermentation broth of fibroin group B; add 8 parts by weight of water and Lactobacillus brevis with a viable count of 10 10 CFU / mL to 1 part by weight of fibroin group C, ferment at 42 °C for 26 h, and sterilize to obtain the fermentation broth of fibroin group C;

[0073] S3. Decolorize the fermentation broth of fibroin group A, fibroin group B, and fibroin group C with activated carbon respectively, and filter through a 0.22 μm microfiltration membrane to obtain the decolorized solution of fibroin group A, the decolorized solution of fibroin group B, and the decolorized solution of fibroin group C respectively;

[0074] S4. Respectively, concentrate and dry the decolorization solutions of fibroin group A, fibroin group B, and fibroin group C at low temperature, and then perform ultraviolet sterilization treatment to obtain the powders of fibroin group A, fibroin group B, and fibroin group C;

[0075] S5. Thoroughly mix the powders of fibroin group A, fibroin group B, and fibroin group C with a weight ratio of 1:1.3:0.6 to obtain hydrolyzed silk.

[0076] The preparation method of the cell active peptide composition with anti-allergic and redness-reducing effects in this example includes the following steps: Weigh the corresponding components according to the weight ratio, mix them evenly, and sterilize to obtain the cell active peptide composition.

[0077] Example 2

[0078] This example provides a cell active peptide composition, including the following components in parts by weight: 1.2 parts of tripeptide-1, 0.5 part of ceramide, 2.8 parts of hydrolyzed silk, 1.9 parts of mussel extract, 0.01 part of sodium hyaluronate, 0.0001 part of 4-tert-butylcyclohexanol, 0.2 part of dipeptide-1, 0.0001 part of palmitoyl tripeptide-8, 0.05 part of palmitoyl tetrapeptide-7, 0.007 part of sh-pentapeptide-1, 0.09 part of dipeptide-2, 0.2 part of bisabolol, 0.12 part of acetyl tetrapeptide-3.

[0079] The preparation method of the mussel extract is as follows:

[0080] (1) Take out the fresh mussel meat from the green-lipped mussel, break the mussel meat, and add 5 times the weight of water of the mussel meat to obtain a mixture;

[0081] (2) Add a complex enzyme to the mixture for enzymatic hydrolysis. Hydrolyze at 30°C for 1 day, and after inactivating the enzyme, obtain the enzymatic hydrolysis product;

[0082] (3) Add a complex bacterium to the enzymatic hydrolysis product for fermentation. Ferment at 38°C for 3 days, and sterilize to obtain the fermentation product;

[0083] (4) First filter the fermentation product through a 0.45 μm filter membrane to obtain a filtrate, then ultrafilter the filtrate through a 10 kDa ultrafiltration membrane to obtain an ultrafiltrate, and perform spray drying to obtain the mussel extract.

[0084] The complex enzyme includes pectinase, bromelain, and chitosanase.

[0085] The enzyme activity of pectinase in the mixture is 70 U / mL; the enzyme activity of bromelain in the mixture is 170 U / mL; the enzyme activity of chitosanase in the mixture is 110 U / mL.

[0086] The composite bacteria include Bifidobacterium longum, Bifidobacterium lactis and Lactobacillus rhamnosus.

[0087] In the enzymolysis product, the viable count of Bifidobacterium longum is 5.0×10 9 CFU / mL, the viable count of Bifidobacterium lactis is 6.0×10 8 CFU / mL, and the viable count of Lactobacillus rhamnosus is 1.5×10 9 CFU / mL.

[0088] The preparation method of hydrolyzed silk is as follows:

[0089] S1. Clean and boil the mulberry cocoon shell to degum it, and obtain fibroin.

[0090] S2. Divide the fibroin into three groups, namely fibroin group A, fibroin group B and fibroin group C; add 10 parts by weight of water and Bifidobacterium breve with a viable count of 10 9 CFU / mL to 1 part by weight of fibroin group A, ferment at 40°C for 50 h, and sterilize to obtain the fermentation broth of fibroin group A; add 10 parts by weight of water and Lactobacillus casei with a viable count of 10 9 CFU / mL to 1 part by weight of fibroin group B, ferment at 40°C for 60 h, and sterilize to obtain the fermentation broth of fibroin group B; add 7 parts by weight of water and Lactobacillus brevis with a viable count of 10 9 CFU / mL to 1 part by weight of fibroin group C, ferment at 40°C for 30 h, and sterilize to obtain the fermentation broth of fibroin group C;

[0091] S3. Decolorize the fermentation broth of fibroin group A, the fermentation broth of fibroin group B and the fermentation broth of fibroin group C with activated carbon respectively, and filter through a 0.22 μm microfiltration membrane to obtain the decolorized solution of fibroin group A, the decolorized solution of fibroin group B and the decolorized solution of fibroin group C respectively;

[0092] S4. Concentrate and dry the decolorized solution of fibroin group A, the decolorized solution of fibroin group B and the decolorized solution of fibroin group C at low temperature respectively, and then perform ultraviolet sterilization treatment to obtain the powder of fibroin group A, the powder of fibroin group B and the powder of fibroin group C;

[0093] S5. Mix the powder of fibroin group A, the powder of fibroin group B and the powder of fibroin group C with a weight ratio of 1:1.2:0.5 evenly to obtain hydrolyzed silk.

[0094] The preparation method of the cell active peptide composition with anti-allergy and redness-relieving effects in this example includes the following steps: Weigh the corresponding components according to the weight ratio, mix them evenly, and sterilize to obtain the cell active peptide composition.

[0095] Comparative Example 1

[0096] The difference between this comparative example and Example 1 is as follows: A cell active peptide composition comprises the following components in parts by weight: 0.5 part of Tripeptide-1, 0.5 part of ceramide, 1.6 parts of hydrolyzed silk, 2.8 parts of mussel extract, 0.003 part of sodium hyaluronate, 0.1 part of 4-tert-butylcyclohexanol, 0.5009 part of Dipeptide-1, 0.05 part of Palmitoyl Tripeptide-8, 0.008 part of Palmitoyl Tetrapeptide-7, 0.12 part of SH-Pentapeptide-1, 0.09 part of Dipeptide-2, 0.52 part of bisabolol, and 0.03 part of Acetyl Tetrapeptide-3.

[0097] Comparative Example 2

[0098] The difference between this comparative example and Example 1 is that the mussel extract is a commercially available product purchased from Xi'an Ruili Biotechnology Co., Ltd.

[0099] Comparative Example 3

[0100] The difference between this comparative example and Example 1 is that the composite enzyme includes neutral protease, ficin, and pectinase.

[0101] The enzyme activity of neutral protease in the mixture is 80 U / mL; the enzyme activity of ficin in the mixture is 160 U / mL; the enzyme activity of pectinase in the mixture is 120 U / mL.

[0102] Comparative Example 4

[0103] The difference between this comparative example and Example 1 is that the composite bacteria include Bifidobacterium breve, Lactobacillus casei, and Lactobacillus brevis.

[0104] The viable count of Bifidobacterium breve in the enzymolysis product is 4.0×10 9 CFU / mL, the viable count of Lactobacillus casei is 7.0×10 8 CFU / mL, and the viable count of Lactobacillus brevis is 1×10 9 CFU / mL.

[0105] Comparative Example 5

[0106] The difference between this comparative example and Example 1 is that the hydrolyzed silk is a commercially available product purchased from Hubei Jianchu Biopharmaceutical Co., Ltd.

[0107] Comparative Example 6

[0108] The difference between this comparative example and Example 1 is that Bifidobacterium breve is replaced by Bifidobacterium longum; Lactobacillus casei is replaced by Bifidobacterium lactis; Lactobacillus brevis is replaced by Lactobacillus rhamnosus.

[0109] Comparative Example 7

[0110] The difference between this comparative example and Example 1 is as follows: Silk fibroin powder in Group A, silk fibroin powder in Group B, and silk fibroin powder in Group C with a weight ratio of 1:1:1 were thoroughly mixed evenly according to a specific weight ratio to obtain hydrolyzed silk.

[0111] Comparative Example 8

[0112] The difference between this comparative example and Example 1 is as follows: A cell-active peptide composition, comprising the following components in parts by weight: 2 parts of Tripeptide-1, 0.5 part of ceramide, 2.2 parts of hydrolyzed silk, 1.7 parts of mussel extract, 0.003 part of sodium hyaluronate, 0.1 part of 4-tert-butylcyclohexanol, 0.0009 part of Dipeptide-1, 0.05 part of Palmitoyl Tripeptide-8, 0.008 part of Palmitoyl Tetrapeptide-7, 0.12 part of SH-Pentapeptide-1, 0.09 part of Dipeptide-2, 0.02 part of bisabolol, 0.03 part of Acetyl Tetrapeptide-3.

[0113] Performance Test

[0114] 1. Anti-allergy experiment: Hyaluronidase activity experiment: Hyaluronic acid is widely present in the skin and other tissues, and the relationship between hyaluronic acid and the skin is very close. Macromolecular hyaluronic acid can inhibit inflammatory reactions, thus alleviating the discomfort caused by skin inflammation. Macromolecular hyaluronic acid will be degraded under the action of hyaluronidase, resulting in a decrease in the content of macromolecular hyaluronic acid, and the degradation product N-acetylglucosamine is produced by decomposition. Measuring N-acetylglucosamine in the reaction system can indirectly reflect the activity of hyaluronidase. The anti-allergy activity of the sample was evaluated by an in vitro experiment for inhibiting hyaluronidase activity. Inhibiting hyaluronidase activity can reduce the degradation of hyaluronic acid and alleviate skin sensitivity.

[0115] Detection method: The cell-active peptide compositions of Examples 1-2 and Comparative Examples 1-8 were respectively mixed with hyaluronidase solution. The concentration of the cell-active peptide composition in the solution was 7 wt%. After shaking and mixing, it was incubated at 37 °C for 10 min. Sodium hyaluronate solution was added and incubated at 37 °C for 45 min. BSA solution was added and incubated for another 10 min, and the OD600nm value was measured to calculate the hyaluronidase inhibition rate. See Table 1.

[0116] Table 1: Hyaluronidase inhibition rate

[0117] Inhibition rate % Example 1 100 Example 2 99 Comparative Example 1 83 Comparative Example 2 85 Comparative Example 3 89 Comparative Example 4 90 Comparative Example 5 87 Comparative Example 6 88 Comparative Example 7 91 Comparative Example 8 96

[0118] 2. Redness reduction effect test

[0119] 5-Lipoxygenase is an enzyme involved in the synthesis of inflammatory mediators. It can initiate the conversion of arachidonic acid to leukotrienes, and leukotrienes are a class of substances related to inflammatory reactions. During the skin inflammation process, an increase in the activity of 5-lipoxygenase will lead to inflammatory symptoms such as skin redness and swelling.

[0120] Sample group: The cell active peptide compositions of Examples 1-2 and Comparative Examples 1-8 were accurately prepared into 1 mg / mL solutions respectively using 100 mmol / L phosphate buffer solution (PBS) with pH 6.9 for standby.

[0121] Substrate solution preparation: Prepare a PBS buffer solution with a concentration of 200 mmol / L and pH 9.0. Accurately pipette 50 μL of polysorbate-20 and disperse it in the PBS buffer solution. While shaking, add 50 μL of linoleic acid. After mixing evenly, add 1 mol / L sodium hydroxide while shaking until the whole system becomes clear. Adjust the pH to 9.0 and make up the volume to 100 mL with PBS;

[0122] 5-Lipoxygenase inhibitory activity assay: Prepare a 5-lipoxygenase solution with a concentration of 0.2 U / mL using PBS. Accurately pipette 50 μL of the sample solution into a 96-well plate, add 100 μL of the substrate solution, gently shake and mix well, incubate at room temperature for 5 min, quickly add 100 μL of the enzyme solution, measure the absorbance A value (referring to the absorbance of the microplate reader at different wavelengths) at 234 nm of the microplate reader at 0 and 5 min of the reaction respectively. Set up a sample group (the concentration of each sample component is 0.2 mg / mL) and a blank group (PBS instead of the sample), record the difference in A for 2 times, perform parallel operations 3 times, and calculate the inhibition rate according to the following formula: Inhibition rate = (△A0 - △A) / △A0, where △A0 is the difference in A of the blank group, and △A is the difference in A of the sample group. The specific detection results are shown in Table 2 below.

[0123] Table 2: Comparison table of 5-lipoxygenase inhibition rates

[0124]

[0125]

[0126] 3. Storage stability

[0127] The cell active peptide compositions prepared in Examples 1-2 and Comparative Examples 1-8 were sealed and stored, and were respectively placed in the following 4 environments for observation to test the storage stability.

[0128] Normal temperature: The samples were left standing at 25 °C and 60% RH (relative humidity) for 180 days.

[0129] Cold resistance: Placed at -10 °C for 30 days.

[0130] Thermal cycling: -10 °C, 25% RH; 25 °C, 60% RH; 38 °C, 75% RH, 24 h for each temperature, a total of 30 cycles.

[0131] High-temperature acceleration: The test conditions were 50 °C ± 1 °C and 25% RH ± 5% RH for 90 days.

[0132] No obvious change is recorded as passing, and the appearance of odor and / or color change is recorded as failing.

[0133] The results are shown in Table 3.

[0134] Table 3: Results of Stability Test

[0135]

[0136]

[0137] As can be seen from the above performance test results, the cell-active peptide compositions of Examples 1-2 have good anti-allergic and anti-reddening effects. In particular, the comprehensive performance of Example 1 is the most prominent, which is mainly due to the synergistic compounding of multiple components.

[0138] In the comparative examples, since the necessary technical solutions were not adopted, their performance in the corresponding performance tests was significantly worse than that of the examples. In Comparative Example 1, the component ratio was changed, and it can be seen that the comprehensive effect decreased. In Comparative Example 2, a commercially available mussel extract was used. In Comparative Examples 3-4, the scheme for preparing the mussel extract was not used, and it can be seen from the results that the anti-allergic effect was affected. In Comparative Example 5, a commercially available hydrolyzed silk was used. In Comparative Examples 6-7, the scheme for preparing the hydrolyzed silk was not used, and the anti-reddening effect of the product decreased. In Comparative Example 8, the ratio of the mussel extract, hydrolyzed silk, and Tripeptide-1 was changed, and the storage stability of the product decreased. The above experimental results further prove the importance of the technical solutions defined in the present invention for its technical effects.

[0139] The above is the preferred embodiment of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A cell active peptide composition with anti-allergic and redness-removing effects, characterized in that: The invention comprises the following components in parts by weight: 0.0001-2 parts of tripeptide-1, 0.0001-0.8 parts of ceramide, 0.0001-3 parts of hydrolyzed silk, 0.0001-2.6 parts of mussel extract, 0.0001-0.2 parts of sodium hyaluronate, 0.0001-1 parts of 4-tert-butyl cyclohexanol, 0.0001-0.2 parts of dipeptide-1, 0.0001-0.2 parts of palmitoyl tripeptide-8, 0.0001-0.2 parts of palmitoyl tetrapeptide-7, 0.0001-0.15 parts of sh-pentapeptide-1, 0.0001-0.15 parts of dipeptide-2, 0.0001-0.2 parts of bisabolol, and 0.0001-0.15 parts of acetyl tetrapeptide-3; The preparation method of the mussel extract is: (1) taking fresh mussel meat from green-lipped mussels, crushing the mussel meat, and adding water 5 to 6 times the weight of the mussel meat to obtain a mixture; (2) adding a composite enzyme to the mixture for enzymolysis at 30-35° C. for 1-2 days, inactivating the enzyme, and obtaining an enzymolysis product; (3) adding composite bacteria to the enzymatic hydrolyzate for fermentation at 38-40° C. for 3-5 days, sterilizing, and obtaining a fermentation product; (4) filtering the fermentation product using a 0.45 μm filter membrane to obtain a filtrate, ultrafiltering the filtrate using a 10 kDa ultrafiltration membrane to obtain an ultrafiltrate, and then spray drying to obtain a mussel extract; The complex enzyme comprises pectinase, bromelain and chitosanase; the enzyme activity of pectinase in the mixture is 70U / mL-90U / mL; the enzyme activity of bromelain in the mixture is 150U / mL-170U / mL; the enzyme activity of chitosanase in the mixture is 110U / mL-130U / mL; The composite bacteria include Bifidobacterium longum, Bifidobacterium lactis and Lactobacillus rhamnosus; the number of live Bifidobacterium longum in the enzymatic hydrolysis product is 3.0×10 9 CFU / mL-5.0×10 9 CFU / mL, number of live Bifidobacterium lactis 6.0×10 8 CFU / mL-8.0×10 8 CFU / mL, the number of live Lactobacillus rhamnosus cells is 0.5×10 9 CFU / mL-1.5×10 9 CFU / mL; The preparation method of hydrolyzed silk is as follows: S1, washing and boiling silkworm cocoon shells to degumming to obtain silk fibroin; S2, the silk fibroin is divided into three groups, namely, silk fibroin group A, silk fibroin group B and silk fibroin group C; 7-10 parts by weight of water and 10 parts by weight of bacterial activity in the system are added to 1 part by weight of silk fibroin group A. 9 CFU / mL-10 10 CFU / mL of breve Bifidobacterium, ferment at 40-45°C for 40-50h, sterilize, and obtain a fermentation liquid of silk fibroin group A; add 7-10 parts by weight of water and 10 parts by weight of bacteria active in the system to 1 part by weight of silk fibroin group B; 9 CFU / mL-10 10 CFU / mL of Lactobacillus casei, fermented at 40-45°C for 60-70h, sterilized, and obtained a silk fibroin group B fermentation liquid; 1 part by weight of silk protein C group is added with 7-10 parts by weight of water and 10 parts by weight of bacteria in the system. 9 CFU / mL-10 10 CFU / mL of Lactobacillus brevis, ferment at 40-45°C for 20-30h, sterilize, and obtain the silk fibroin group C fermentation liquid; S3, decolorizing the silk fibroin group A fermentation liquid, the silk fibroin group B fermentation liquid and the silk fibroin group C fermentation liquid respectively through activated carbon, and filtering through a 0.22 μm micro-membrane to obtain a silk fibroin group A decolorizing liquid, a silk fibroin group B decolorizing liquid and a silk fibroin group C decolorizing liquid respectively; S4, respectively concentrating and drying the silk fibroin group A decolorizing solution, the silk fibroin group B decolorizing solution and the silk fibroin group C decolorizing solution at low temperature, and then sterilizing them with ultraviolet light to obtain silk fibroin group A powder, silk fibroin group B powder and silk fibroin group C powder; S5, mixing the silk fibroin group A powder, the silk fibroin group B powder and the silk fibroin group C powder in a specific weight ratio to obtain hydrolyzed silk; In the step S5, the weight ratio of the silk fibroin group A powder, the silk fibroin group B powder and the silk fibroin group C powder is 1:(1.2-1.4):(0.5-0.8).

2. The cell active peptide composition according to claim 1, characterized in that: The weight ratio of mussel extract, hydrolyzed silk and tripeptide-1 is (1.2-1.5):(2.4-2.8):(1.9-2.2).

3. A method for preparing the cell active peptide composition according to any one of claims 1 to 2, characterized in that: The corresponding components are weighed according to the weight ratio, mixed evenly, and sterilized to obtain the cell active peptide composition.

4. Use of the cell active peptide composition according to any one of claims 1 to 2 in the preparation of skin products.

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