CN117481996A - Mild anti-aging composition containing recombinant collagen and application thereof - Google Patents
Mild anti-aging composition containing recombinant collagen and application thereof Download PDFInfo
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- CN117481996A CN117481996A CN202311622947.1A CN202311622947A CN117481996A CN 117481996 A CN117481996 A CN 117481996A CN 202311622947 A CN202311622947 A CN 202311622947A CN 117481996 A CN117481996 A CN 117481996A
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- recombinant collagen
- aging
- mild
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- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
- A61K8/65—Collagen; Gelatin; Keratin; Derivatives or degradation products thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/36—Carboxylic acids; Salts or anhydrides thereof
- A61K8/365—Hydroxycarboxylic acids; Ketocarboxylic acids
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/67—Vitamins
- A61K8/671—Vitamin A; Derivatives thereof, e.g. ester of vitamin A acid, ester of retinol, retinol, retinal
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
- A61K8/735—Mucopolysaccharides, e.g. hyaluronic acid; Derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9728—Fungi, e.g. yeasts
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
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Abstract
The invention relates to a mild anti-aging composition containing recombinant collagen and application thereof. The invention creatively combines two components of vitamin A alcohol and glycollic acid, and simultaneously combines the recombinant collagen to jointly play the anti-aging effect. Meanwhile, in consideration of the fact that retinol and glycolic acid have certain irritation to skin, the combination of the recombinant collagen, the schizophyllan and the chondroitin sulfate is taken as an anti-irritation component, the three components have remarkable synergistic effect in the aspect of reducing skin irritation, and the composition can well give consideration to excellent anti-aging effect and excellent mildness and has a prospect in the application of the composition to the preparation of cosmetics.
Description
Technical Field
The invention belongs to the technical field of cosmetics, and relates to a mild anti-aging composition containing recombinant collagen and application thereof.
Background
Aging is also known as aging, which is an essential stage in the course of life activities, and skin aging is most evident in the aging process of the body. Skin aging not only causes wrinkles, looseness and pigmentation, but also causes problems such as reduced metabolism of epidermis, impaired ability of epidermis repair and impaired ability of barrier protection. The composition has a plurality of anti-aging effects, but the composition has high efficiency and mildness. For example, hydroxy acid based compositions can improve the appearance of photoaged or naturally aged skin, but can be irritating to the skin, and can cause varying degrees of redness or stinging after use.
With the vigorous development of cosmetic markets, consumers have demands for not only anti-aging, whitening and other effects, but also increasing demands for mildness and nonirritating skin care products. The Chinese cosmetic market is the biggest emerging market worldwide, and a great deal of skin care cosmetics are emerging to make people feel at the spotlight, so that people not only learn to use good cosmetics, but also pay more attention to the effect and safety of the cosmetics.
Factors affecting skin irritation include intrinsic factors such as race differences, age, skin health, and external factors such as climate, and cosmetic irritation components. Skin irritation may cause damage to the skin barrier, abnormal nerve function, inflammatory response, etc. For people with sensitive skin, skin irritation may cause the skin to be drier, easy to tighten and redder, even slight irritation can cause itching, desquamation, burning, stinging and other sensations, and serious people generate bad emotion or mental symptoms.
It would therefore be of great interest to be able to develop a product that combines both efficient anti-ageing and gentle non-irritating effects.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a mild anti-aging composition containing recombinant collagen and application thereof.
In order to achieve the aim of the invention, the invention adopts the following technical scheme:
in a first aspect, the present invention provides a mild anti-aging composition comprising recombinant collagen, the mild anti-aging composition comprising tretinoin, glycolic acid, recombinant collagen, schizophyllan, and chondroitin sulfate.
Vitamin a alcohol is a fat-soluble vitamin consisting of 3, 7-dimethoxy vitamin a derivative, an active form of vitamin a. The vitamin A alcohol can promote collagen synthesis, promote skin metabolism, increase skin elasticity, and prevent and reduce wrinkle.
The glycollic acid can permeate deep skin, remove cutin, gently dissolve dead skin cells, improve skin texture, color and pore size, and also improve the level of collagen and elastin in skin, thereby helping to resist wrinkles.
The invention creatively combines two components of the vitamin A alcohol and the glycollic acid, and simultaneously combines the recombinant collagen to jointly play the anti-aging effect, and the three components have potential interaction in the aspect of anti-aging, namely, the vitamin A alcohol and the glycollic acid and the recombinant collagen have synergistic interaction. Meanwhile, in consideration of the fact that retinol and glycolic acid have certain irritation to skin, the combination of the recombinant collagen, the schizophyllan and the chondroitin sulfate is taken as an anti-irritation component, the three components have remarkable synergistic effect in the aspect of reducing skin irritation, and the composition can well give consideration to excellent anti-aging effect and excellent mildness and has a prospect in the application of the composition to the preparation of cosmetics.
Preferably, the mild anti-aging composition comprises, by mass, 0.1-0.5 part of tretinoin, 0.5-2 parts of glycolic acid, 0.01-1 part of recombinant collagen, 0.1-0.5 part of schizophyllan polysaccharide and 0.1-3 parts of chondroitin sulfate.
Based on the potential interaction between the components, when the five components are combined in a specific mass ratio relationship, the five components have better effects on reducing skin irritation and improving skin aging resistance.
The weight parts of the vitamin A alcohol can be selected from 0.1 part, 0.15 part, 0.2 part, 0.25 part, 0.3 part, 0.35 part, 0.4 part, 0.45 part, 0.5 part and the like; the mass parts of the glycollic acid can be selected from 0.5 part, 0.6 part, 0.7 part, 0.8 part, 1 part, 1.2 part, 1.5 part, 1.8 part, 2 parts and the like; the weight parts of the recombinant collagen can be selected from 0.01 part, 0.03 part, 0.05 part, 0.08 part, 0.1 part, 0.2 part, 0.4 part, 0.5 part, 0.8 part, 1 part and the like; the mass parts of the carboxymethyl schizophyllan can be selected to be 0.1 part, 0.15 part, 0.2 part, 0.25 part, 0.3 part, 0.35 part, 0.4 part, 0.45 part, 0.5 part and the like; the mass part of the chondroitin sulfate can be selected from 0.1 part, 0.2 part, 0.3 part, 0.5 part, 0.8 part, 1 part, 1.5 part, 2 parts, 2.5 parts, 3 parts and the like; other specific point values in the numerical ranges are selectable, and will not be described in detail herein.
Preferably, the nucleotide sequence encoding the recombinant collagen is the sequence shown in SEQ ID NO. 1.
Compared with other sources of recombinant collagen, the recombinant collagen from the specific source has better effects on supplementing skin collagen, resisting oxidation and aging and reducing skin irritation.
SEQ ID NO:1：
GGGCCTCAAGGTATTGCTGGACAGCGTGGTGTGGTCGGCCTGCCTGGTCAGAGAGGAGAGAGAGGCTTCCCTGGTCTTCCTGGCCCCTCTGGTGAACCTGGCAAACAAGGTCCCTCTGGAGCAAGTGGTGAACGTGGTCCCCCTGGTCCCATGGGCCCCCCTGGATTGGCTGGACCCCCTGGTGAATCTGGACGTGAGGGGGCTCCTGGTGCCGAAGGTTCCCCTGGACGAGACGGTTCTCCTGGCGCCAAGGGTGACCGTGGTGAGACCGGCCCCGCTGGACCCCCTGGTGCTCCTGGTGCTCCTGGTGCCCCTGGCCCCGTTGGCCCTGCTGGCAAGAGTGGTGATCGTGGTGAGACTGGTCCTGCTGGTCCCGCCGGAGAACGAGGTGGCCCTGGAGGACCTGGCCCTCAGGGTCCTCCTGGAAAGAATGGTGAAACTGGACCTCAGGGACCCCCAGGGCCTACTGGGCCTGGTGGTGACAAAGGAGACACAGGACCCCCTGGTCCACAAGGATTACAAGGCTTGCCTGGTACAGGTGGTCCTCCAGGAGAAAATGGAAAACCTGGGGAACCAGGTCCAAAGGGTGATGCCGGTGCACCTGGAGCTCCAGGAGGCAAGGGTGATGCTGGTGCCCCTGGTGAACGTGGACCTCCTGGATTGGCAGGGGCCCCAGGACTTAGAGGTGGAGCTGGTCCCCCTGGTCCCGAAGGAGGAAAGGGTGCTGCTGGTCCTCCTGGGCCACCTGGTGCTGCTGGTACTCCTGGTCTGCAAGGAATGCCTGGACCTGGTCCTTGCTGTGGTGGTTAA。
In a second aspect, the present invention provides the use of a mild anti-ageing composition comprising recombinant collagen according to the first aspect for the preparation of a cosmetic.
Preferably, the weight percentage of the mild anti-aging composition containing recombinant collagen in the cosmetic is 0.1-10%, for example 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 7%, 8%, 9%, 10%, etc., and other specific values within the numerical range can be selected, which will not be described in detail herein.
In a third aspect, the invention provides an essence emulsion with an anti-aging effect, wherein the essence emulsion with the anti-aging effect is prepared from the following raw materials in percentage by mass: the mild anti-aging composition containing recombinant collagen according to the first aspect comprises 0.1-10%, emulsifier 1-5%, thickener 0.1-2%, polyalcohol 5-20% and water 50-95%.
The mild anti-aging composition may be 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 7%, 8%, 9%, 10%, etc.; the mass percentage of the emulsifying agent can be 1%, 2%, 3%, 4%, 5% and the like; the mass percentage of the thickener can be 0.1%, 0.5%, 0.7%, 0.8%, 1%, 1.2%, 1.4%, 1.5%, 1.6%, 1.8%, 2% and the like; the mass percent of the polyol can be 5%, 6%, 8%, 10%, 12%, 14%, 15%, 17%, 18%, 19%, 20%, etc.; the water may be 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, etc.; other specific point values in the numerical ranges are selectable, and will not be described in detail herein.
Preferably, the emulsifier comprises any one or a combination of at least two of potassium cetyl phosphate, polysorbate-60, methyl glucose sesquistearate, sorbitan stearate, sorbitol polyether-30 tetraoleate, polysorbate-20, cetostearyl olivate/sorbitan olivate or cetyl PEG/PPG-10/1 polydimethylsiloxane.
Preferably, the thickener comprises any one or a combination of at least two of sodium polyacrylate, sodium polyacrylate grafted starch, ammonium acryloyldimethyl taurate/VP copolymer or xanthan gum.
Preferably, the polyol comprises any one or a combination of at least two of glycerol, 1, 3-propanediol, panthenol, 1, 2-hexanediol or butanediol.
Preferably, the preparation raw materials of the essence milk with the anti-aging effect further comprise the following components in percentage by mass: 1-10% of plant source conditioning agent, 0.1-2% of pH regulator, 0.01-0.5% of chelating agent, 0.05-0.5% of antioxidant and 0.1-0.5% of aromatic agent or the combination of at least two.
The mass percentage of the plant source conditioning agent can be selected to be 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% and the like; the mass percentage of the pH regulator can be selected to be 0.1%, 0.3%, 0.5%, 0.8%, 1%, 1.2%, 1.4%, 1.6%, 1.8%, 2% and the like; the mass percentage of the chelating agent can be selected to be 0.01%, 0.03%, 0.05%, 0.08%, 0.1%, 0.15%, 0.2%, 0.3%, 0.4%, 0.5% and the like; the antioxidant may be selected from 0.05%, 0.08%, 0.1%, 0.15%, 0.2%, 0.3%, 0.4%, 0.5%, etc.; the mass percentage of the aromatic agent can be selected to be 0.1%, 0.15%, 0.2%, 0.3%, 0.4%, 0.5% and the like; other specific point values in the numerical ranges are selectable, and will not be described in detail herein.
Preferably, the plant source conditioner is selected from any one or a combination of at least two of perilla leaf extract, silybum marianum extract and lentinus edodes extract; preferably, the combination of perilla leaf extract, silybum marianum extract and shiitake mushroom extract.
When the composition is used for preparing cosmetics, the perilla leaf extract, the silybum marianum extract and the lentinus edodes extract are further added, so that the effect of relieving and resisting irritation of the product can be further improved.
In a fourth aspect, the present invention provides a method for preparing the essence milk with anti-aging effect according to the third aspect, the method comprising:
mixing water and part of polyalcohol in a water phase pot, heating to 60-80deg.C, maintaining the temperature for 10-40min, cooling to 25-40deg.C, and mixing with vitamin A alcohol, glycolic acid, recombinant collagen and chondroitin sulfate to obtain water phase mixture;
mixing the emulsifier, the thickener, the rest polyol, the schizophyllan and the rest raw materials in an emulsifying pot, homogenizing for 5-15min at 30-50 ℃, then mixing with the water phase mixture, and homogenizing for 5-30min at 25-40 ℃ to obtain the essence emulsion with anti-aging effect.
The specific point value in 60-80deg.C can be 60 deg.C, 65 deg.C, 70 deg.C, 75 deg.C, 80 deg.C, etc.; the specific point value in the 10-40min can be selected from 10min, 15min, 20min, 25min, 30min, 35min, 40min, etc.; the specific point value of 25-40deg.C can be selected from 25deg.C, 30deg.C, 35deg.C, 40deg.C, etc.; the specific point value of 30-50deg.C can be 30 deg.C, 32 deg.C, 33 deg.C, 35 deg.C, etc.; the specific point value in the 5-15min can be selected from 5min, 6min, 7min, 8min, 10min, 13min, 15min, etc.; the specific point value in the 5-30min can be selected from 5min, 6min, 7min, 8min, 10min, 13min, 15min, 20min, 25min, 30min, etc.; other specific point values in the numerical ranges are selectable, and will not be described in detail herein.
Compared with the prior art, the invention has the following beneficial effects:
the invention creatively combines two components of the vitamin A alcohol and the glycollic acid, and simultaneously combines the recombinant collagen to jointly play the anti-aging effect, and the three components have potential interaction in the aspect of anti-aging, namely, the vitamin A alcohol and the glycollic acid and the recombinant collagen have synergistic interaction. Meanwhile, in consideration of the fact that retinol and glycolic acid have certain irritation to skin, the combination of the recombinant collagen, the schizophyllan and the chondroitin sulfate is taken as an anti-irritation component, the three components have remarkable synergistic effect in the aspect of reducing skin irritation, and the composition can well give consideration to excellent anti-aging effect and excellent mildness and has a prospect in the application of the composition to the preparation of cosmetics.
Detailed Description
In order to further describe the technical means adopted by the present invention and the effects thereof, the following describes the technical scheme of the present invention in combination with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.
The mass parts of each component in the composition product referred to in the following are calculated according to the actual content of the effective components in the raw materials sold in the market.
The schizophyllan polysaccharide involved in the following is derived from the product of the Chengsu biosciences of Shaanxi, inc. with the model TZCS W02905; chondroitin sulfate is derived from the product of guangzhou Hua biosciences limited; the perilla leaf extract is derived from the product of Waters biotechnology Co., ltd; the silybum marianum extract is derived from the product of the company of the biological technology of Volterras, lanzhou; the Lentinus edodes extract is derived from the product of Waters biotechnology Co., ltd; sodium polyacrylate is a product from guangzhou Tianjia biotechnology limited company; xanthan gum is derived from Shanghai spectral vibro-biological limited.
The recombinant collagen referred to in the following was prepared using the product prepared in preparation examples in which pET-28a (+) vector was purchased from Michelin; BL21 (DE 3) chemocompetent cells were commercially available from Merck under the designation CMC0014-4X40 UL.
Preparation example
The preparation example provides recombinant collagen freeze-dried powder, and the preparation method is carried out by referring to the method disclosed in the preparation example 1 in ZL 202310123476.3, and specifically comprises the following steps:
(1) Constructing a recombinant plasmid: according to the nucleotide sequence shown in SEQ ID NO. 1, selecting a pET-28a (+) vector suitable for an expression vector escherichia coli host, and sending the pET-28a (+) vector to a sequencing company for synthesis to prepare a recombinant collagen expression vector pET-28a-1.
(2) Recombinant fermentation strain construction: transferring the pET-28a-1 expression vector into competent cells of escherichia coli (BL 21 (DE 3)) by using a chemical conversion method, screening positive clones in plate colonies after conversion by agarose gel electrophoresis, transferring the screened positive clones to 2mL of LB culture medium, culturing for 12 hours at 37 ℃ and 200rpm, diluting the cultured bacterial liquid by 1 time with glycerol with the volume fraction of 50%, packaging into a freezing tube, and freezing at the temperature of minus 80 ℃ to obtain the recombinant collagen fermentation strain.
(3) Fermenting and culturing recombinant fermentation strains: 10 mu L of recombinant collagen fermentation broth is coated on a solid LB plate, cultured for 12 hours in a 37 ℃ incubator, selected and inoculated in 50mL of LB liquid medium containing 50 mu g/mL kanamycin content, placed in a shaking table for culture at 200rpm for 14 hours at 37 ℃, then inoculated in 200mL of LB liquid medium containing 50 mu g/mL kanamycin content at a ratio of 1:100, cultured until the bacterial broth OD=0.6 at 37 ℃, added with IPTG to achieve the final concentration of 1mM, continuously cultured for 4 hours, then centrifuged at 4000rpm for 15 minutes, and the bacterial cells are collected.
(4) Purifying recombinant collagen: the bacterial cells are subjected to ultrasonic disruption and centrifugation, the precipitate is washed by using a TritonX-100 washing solution, the supernatant is removed by centrifugation again, 6M guanidine hydrochloride is added into the precipitate, the pH is regulated to 8, after the precipitate is fully dissolved, the solution is subjected to nickel affinity chromatography resin, 20mM, 50mM, 100mM, 200mM and 500mM concentration gradient imidazole solution is used for eluting in sequence, eluent is concentrated by using a protein concentration column, the concentrated solution is collected and dialyzed overnight (dialysis bag, RC film, 14KD and the flat width is 44mM (Producer F600112)), then the protein concentration is detected by using a BCA kit, and the solution is uniformly mixed with the mannitol aqueous solution according to the proportion of 0.5mL of 5% mannitol aqueous solution by volume concentration per 5mg protein, and then the solution is pre-frozen at-40 ℃ for 10min by using a freeze dryer, and finally freeze-dried at-35 ℃ to obtain the recombinant collagen freeze-dried powder.
Example 1
The embodiment provides a mild anti-aging composition which comprises the following components in parts by weight: 0.2 part of vitamin A alcohol, 0.7 part of glycollic acid, 0.05 part of recombinant collagen, 0.2 part of schizophyllan and 0.3 part of chondroitin sulfate.
Example 2
The embodiment provides a mild anti-aging composition which comprises the following components in parts by weight: 0.1 part of vitamin A alcohol, 1 part of glycollic acid, 0.01 part of recombinant collagen, 0.5 part of schizophyllan and 1 part of chondroitin sulfate.
Example 3
The embodiment provides a mild anti-aging composition which comprises the following components in parts by weight: 0.4 part of vitamin A alcohol, 0.5 part of glycollic acid, 0.05 part of recombinant collagen, 0.1 part of schizophyllan and 0.4 part of chondroitin sulfate.
Comparative example 1
This comparative example provides a composition which differs from example 1 only in that no tretinoin alcohol is contained, the parts by mass of the glycolic acid is 0.9 parts, and the other components and the content remain unchanged.
Comparative example 2
This comparative example provides a composition which differs from example 1 only in that no glycolic acid is present, the parts by weight of tretinoin is 0.9 parts, and the other components and contents remain unchanged.
Comparative example 3
This comparative example provides a composition which differs from example 1 only in that it does not contain recombinant collagen, and its parts by mass are split onto the mass of glycolate, with the other components and the contents remaining unchanged.
Comparative example 4
This comparative example provides a composition differing from example 1 only in that schizophyllan is not contained, the mass fraction of chondroitin sulfate is 0.5, and the other components and contents remain unchanged.
Comparative example 5
This comparative example provides a composition differing from example 1 only in that chondroitin sulfate is not contained, the schizophyllan polysaccharide is 0.5 part by mass, and the other components and contents remain unchanged.
Comparative example 6
This comparative example provides a composition differing from example 1 only in that recombinant collagen is not contained, and its mass fraction is divided over the mass of schizophyllan, and the other components and contents remain unchanged.
Application example 1
The application example provides an essence emulsion, which comprises the following components:
the preparation method comprises the following steps:
(1) Mixing water and glycerol in a water phase pot, heating to 70deg.C, maintaining the temperature for 20min, cooling to 35deg.C, and mixing with vitamin A alcohol, glycolic acid, recombinant collagen and chondroitin sulfate in the composition to obtain water phase mixture;
(2) Mixing emulsifier, thickener, residual polyol and schizophyllan polysaccharide in an emulsifying pot, homogenizing for 10min at 45deg.C, mixing with the water phase mixture, homogenizing for 20min at 35deg.C, and regulating pH to obtain essence emulsion.
Application examples 2 to 3
The present application example provides two kinds of essence milk, the formulation of which is different from that of application example 1 only in that the mild anti-aging composition of example 1 is replaced with the same amount of the anti-aging compositions prepared in examples 2 to 3, and the other components and contents remain unchanged.
The preparation method comprises the following steps:
(1) Mixing water and glycerol in a water phase pot, heating to 65deg.C, maintaining the temperature for 30min, cooling to 30deg.C, and mixing with vitamin A alcohol, glycolic acid, recombinant collagen and chondroitin sulfate in the composition to obtain water phase mixture;
(2) Mixing emulsifier, thickener, residual polyol and schizophyllan polysaccharide in an emulsifying pot, homogenizing for 15min at 40 ℃, then mixing with the water phase mixture, homogenizing for 25min at 30 ℃, and regulating pH value to obtain the essence milk.
Comparative application examples 1 to 6
Comparative examples 1 to 6 provided six kinds of essential milks, the formulation of which was different from that of example 1 only in that the mild anti-aging composition of example 1 was replaced with the same amount of the compositions prepared in comparative examples 1 to 6, and the other components and contents were kept unchanged. The preparation method is described in application example 1.
Application example 4
The application example provides an essence emulsion, which comprises the following components:
the preparation method comprises the following steps:
(1) Mixing water and glycerol in a water phase pot, heating to 70deg.C, maintaining the temperature for 20min, cooling to 35deg.C, and mixing with vitamin A alcohol, glycolic acid, recombinant collagen and chondroitin sulfate in the composition to obtain water phase mixture;
(2) Mixing emulsifier, thickener, residual polyol and schizophyllan polysaccharide in an emulsifying pot, homogenizing for 10min at 45deg.C, mixing with the water phase mixture and plant extract, homogenizing for 20min at 35deg.C, and regulating pH to obtain essence emulsion.
Application example 5
The application example provides an essence emulsion, the formula of which is different from that of application example 4 only in that the perilla leaf extract is lacked, the quality of the essence emulsion is proportionally distributed on the quality of the silybum marianum extract and the shiitake mushroom extract, and other combinations and contents are kept unchanged.
Application example 6
The application example provides an essence emulsion, the formula of which is different from that of application example 4 only in that the silybum marianum extract is absent, the mass of the milk is proportionally distributed on the mass of the perilla leaf extract and the shiitake mushroom extract, and other combinations and contents are kept unchanged.
Application example 7
The application example provides essence milk, the formula of the essence milk is different from that of application example 4 only in that the shiitake mushroom extract is lacked, the quality of the shiitake mushroom extract is distributed to the quality of the silybum marianum extract and the quality of the perilla leaf extract in proportion, and other combinations and contents are kept unchanged.
Test example 1
Chick embryo chorioallantoic membrane vascular assay:
(1) Experimental conditions
Chicken embryo: SPF-grade white Hangzhou chicken.
Culture conditions: the incubator temperature is 37.5+/-0.5 ℃ and the relative humidity is 55-70%.
Reagent and control: positive control (PC 1): 1.0% SDS solution;
positive control (PC 2): 0.1mol/LNaOH solution;
negative Control (NC): 0.9% nacl solution;
test sample (TA): the products prepared in examples 1-3 and comparative examples 1-6 were diluted to 5% with deionized water.
(2) Experimental method
CAM preparation: purchasing a 0 d-old chick embryo, turning once a day in the hatching process, checking the development state of the chick embryo when hatching is 5d years, and stripping eggshell parts to expose white egg membranes when hatching is 6 days; the intima was carefully removed with forceps, ensuring that the vessel membrane was not damaged.
Formal test: each group of 6 chick embryos is prepared by crushing the test substances into powder, and the powder is lightly extruded in a graduated container (such as a microcentrifuge tube) to form 0.3mL volume, directly acts on the CAM to ensure that at least 50% of the surface of the CAM is covered by the test substances, lightly washes the test substances on the CAM membrane with physiological saline after 3min of action, and the washing operation can cover the slight bleeding change on the CAM membrane quickly, so that the result is observed after 30s of washing is finished, the degree of each toxic effect change is observed, and the degree of each toxic effect change is accurate to 3 reactions including bleeding, coagulation and vascular thawing is recorded. The positive and negative controls used the same procedure.
Data analysis: the severity of each response was recorded, graded and scored, and bleeding was classified as no bleeding (0 score), mild bleeding (1 score), moderate bleeding (2 scores), severe bleeding (3 scores) depending on severity; coagulation is classified according to severity as no coagulation (0 min), mild coagulation (1 min), moderate coagulation (2 min), severe coagulation (3 min); vessel thawing is classified into avascular thawing (0 minutes), mild vessel thawing (1 minute), moderate vessel thawing (2 minutes), and severe vessel thawing (3 minutes) according to severity.
Endpoint Scores (ES) were calculated according to the following formula, with the result remaining two bits after the decimal point: score per chick embryo = sum of extent of bleeding, coagulation and vascular thawing observed per chick embryo; es=mathematical sum of 6 chick embryo scores. The eye irritation of the subject was classified according to the following table based on the ES values.
| Endpoint scoring | Irritation classification |
| 0＜ES＜6 | No irritation |
| 6≤ES≤12 | Light irritation |
| 12＜ES＜16 | Moderate irritation |
| ES≥16 | Strong irritation/corrosiveness |
The samples tested were rated for ES values at the end point and the results are shown in Table 1.
TABLE 1
| Group of | Endpoint scoring | Group of | Endpoint scoring |
| Positive control 1 | 18 | Positive control 2 | 18 |
| Negative control | 0 | Example 1 | 3 |
| Example 2 | 4 | Example 3 | 4 |
| Comparative example 1 | 3 | Comparative example 2 | 8 |
| Comparative example 3 | 13 | Comparative example 4 | 11 |
| Comparative example 5 | 9 | Comparative example 6 | 11 |
As can be seen from the data in table 1, the compositions according to the present invention were classified as non-irritating, and the recombinant collagen, schizophyllan and chondroitin sulfate were able to effectively reduce the irritation of the compositions, compared to the results of comparative examples 4-6, and there was a significant synergy between schizophyllan and chondroitin sulfate, and between them and recombinant collagen.
Test example 2
Hyaluronidase in vitro inhibition assay:
hyaluronic acid is a component in tissue matrix which limits diffusion of water and other extracellular substances, and after hydrolysis by hyaluronidase, the cell becomes non-adhesive, so that cell degranulation and synthetic new medium exudation are caused, biological effect is exerted, and rapid allergic reaction is caused. Hyaluronidase is a participant of allergic reaction, has strong correlation with allergy, and can evaluate the anti-allergic effect of the product through the inhibition activity of the hyaluronidase.
The in vitro anti-sensitization efficacy of examples 1-3 and comparative examples 1-6 was examined using the hyaluronidase inhibition method. The products prepared in examples 1 to 3 and comparative examples 1 to 6 were diluted with deionized water to prepare solutions having a concentration of 5% as sample solutions, respectively.
0.1mL CaCl was taken 2 The solution (0.25 mmol/L) and 0.5mL hyaluronidase solution (100U/mL) were reacted in a water bath at 37℃for 20min; adding 0.5mL of the extracting solution, and preserving heat for 20min; then 0.5mL of sodium hyaluronate solution (0.5 g/L) is added, and the mixture is taken out and cooled for 5min after 30min of reaction; 0.1mL of sodium hydroxide solution (0.4 mol/L) and 0.5mL of acetylacetone solution (3.5 mL of acetylacetone was dissolved in 50mL of 1.0mol/L sodium carbonate solution) were added, and immediately after 15min of boiling water bath, the solution was transferred to ice water bath for 5min; 1mL of an Escherichia reagent (0.8 g of p-dimethylaminobenzaldehyde was dissolved in 15mL of concentrated hydrochloric acid and 15mL of absolute ethanol) was added dropwise, and the mixture was diluted with 3mL of absolute ethanol, left at 25℃for 20min for color development, and the absorbance thereof was measured at 540nm with dipotassium glycyrrhizinate as a control.
The inhibition rate for hyaluronidase was calculated according to the following formula:
hyaluronidase inhibition ratio = { [ (A-B) - (C-D) ]/(A-B) } ×100%
Wherein: a—control solution absorbance (with acetate buffer solution instead of sample solution); b—control blank solution absorbance (with acetate buffer solution instead of sample solution and enzyme solution); c-absorbance of sample solution; d-absorbance of sample blank solution (acetic acid buffer solution instead of enzyme solution).
The inhibition rate of hyaluronidase by each group of samples was examined, and the results are shown in table 2:
TABLE 2
| Sample name | Inhibition of hyaluronidase (%) |
| Example 1 | 42.7 |
| Example 2 | 38.4 |
| Example 3 | 40.5 |
| Comparative example 1 | 43.4 |
| Comparative example 2 | 39.6 |
| Comparative example 3 | 21.7 |
| Comparative example 4 | 29.1 |
| Comparative example 5 | 34.3 |
| Comparative example 6 | 25.5 |
As is clear from table 2, the composition of the present invention has a certain inhibitory effect on hyaluronidase, and the inhibition ratio is 38% or more, which indicates that the composition can alleviate or inhibit allergy. And the recombinant collagen, schizophyllan and chondroitin sulfate in the composition have remarkable synergistic effect on the efficacy.
Test example 3
Skin elasticity evaluation test:
(1) The subject: 140 volunteers between the ages of 18 and 50 who were non-physique and hypersensitive (ratio of men to women 1:4) were randomized into 14 groups of 2 men and 8 women each.
Preparation before testing: the inner sides of the forearms of the hands of the subjects are marked with measuring areas, the experimental areas are at least 3cm multiplied by 3cm, and the interval between each measuring area is at least 1cm.
Sample to be tested: the emulsions prepared in application examples 1 to 7 and comparative application examples 1 to 6.
(2) The test process comprises the following steps: the 13 groups of samples are measured according to 2ml/cm 2 Single coating was performed (blank set is pure water). The test specimens were uniformly coated onto the test area using latex gloves and gently tapped until absorbed. Once daily, the skin elasticity test instrument is used for 28 days, the skin elasticity is measured on the 14 th day, after the face is thoroughly cleaned by using a uniform facial cleaning product, the face is gently wiped dry by using a paper towel, the skin elasticity test instrument probe is vertically and lightly touched to the area to be tested after the skin elasticity test instrument probe is left for 15 minutes in a room with the room temperature of 22+/-2 ℃ and the relative humidity of 40+/-2%, and the probe is removed until the test cycle is finished. Each area to be measured was measured at 3 three points, and the results are shown in table 3. On day 28, the anti-wrinkle effect was measured, the face was thoroughly cleaned with a unified cleansing product, gently wiped dry with a paper towel, left standing for 15 minutes in a room with a room temperature of 22.+ -. 2 ℃ and a relative humidity of 40.+ -. 2%, and the face image of the tester was photographed with a VISIA face image analyzer, the analysis area was as full as possibleThe amount avoids interference of the light reflecting area with the hair. Shooting the face (including the front face, the left side face and the right side face) of the subject, carrying out data statistics on the obtained absolute scores, and recording the detection data of wrinkles on the face of the subject. The test results are shown in Table 3.
TABLE 3 Table 3
From the data in Table 3, it is clear that the composition according to the present invention has a good effect of restoring skin elasticity, wherein the glycolic acid and the retinol mutually promote the effect of promoting skin elasticity, and the effect is optimized by acting together with the recombinant collagen.
Test example 4
Sensory evaluation test:
test example 3 the same volunteers, after use, were scored for anti-aging and mildness of the products, respectively, with a score of 5 at the highest and 1 at the lowest, scoring interval 1-5, and the evaluation results are shown in table 4 (average score):
TABLE 4 Table 4
As shown in Table 4, the composition and the cosmetic prepared by the composition have excellent anti-aging property and mildness, the plant extract added into the cosmetic can further improve the mildness effect of the composition, and the perilla leaf extract, the silybum marianum extract and the lentinus edodes extract have a certain synergistic effect on improving the mildness of the product.
The applicant states that the technical solution of the present invention is illustrated by the above embodiments, but the present invention is not limited to the above embodiments, i.e. it does not mean that the present invention must be implemented by the above embodiments. It should be apparent to those skilled in the art that any modification of the present invention, equivalent substitution of raw materials for the product of the present invention, addition of auxiliary components, selection of specific modes, etc., falls within the scope of the present invention and the scope of disclosure.
The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the specific details of the above embodiments, and various simple modifications can be made to the technical solution of the present invention within the scope of the technical concept of the present invention, and all the simple modifications belong to the protection scope of the present invention.
In addition, the specific features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various possible combinations are not described further.
Claims (10)
1. A mild anti-aging composition comprising recombinant collagen, wherein the mild anti-aging composition comprises tretinoin, glycolic acid, recombinant collagen, schizophyllan, and chondroitin sulfate.
2. The mild anti-aging composition containing recombinant collagen according to claim 1, wherein said mild anti-aging composition comprises, in parts by mass, 0.1 to 0.5 part of tretinoin, 0.5 to 2 parts of glycolic acid, 0.01 to 1 part of recombinant collagen, 0.1 to 0.5 part of schizophyllan and 0.1 to 3 parts of chondroitin sulfate.
3. The mild anti-aging composition containing recombinant collagen according to claim 1 or 2, wherein the nucleotide sequence encoding said recombinant collagen is the sequence shown in SEQ ID No. 1.
4. Use of a mild anti-ageing composition comprising recombinant collagen according to any one of claims 1 to 3 for the preparation of a cosmetic.
5. The use according to claim 4, wherein the mild anti-aging composition containing recombinant collagen is present in the cosmetic in an amount of 0.1 to 10% by mass.
6. The essence emulsion with the anti-aging effect is characterized by comprising the following preparation raw materials in percentage by mass: a mild anti-aging composition according to any one of claims 1 to 3, comprising 0.1 to 10% of recombinant collagen, 1 to 5% of an emulsifier, 0.1 to 2% of a thickener, 5 to 20% of a polyol and 50 to 95% of water.
7. The anti-aging cream of claim 6, wherein the emulsifier comprises any one or a combination of at least two of potassium cetyl phosphate, polysorbate-60, methyl glucose sesquistearate, sorbitan stearate, sorbitol polyether-30 tetraoleate, polysorbate-20, cetostearyl oleate/sorbitan olivate or cetyl PEG/PPG-10/1 polydimethylsiloxane;
preferably, the thickener comprises any one or a combination of at least two of sodium polyacrylate, sodium polyacrylate grafted starch, ammonium acryloyldimethyl taurate/VP copolymer or xanthan gum;
preferably, the polyol comprises any one or a combination of at least two of glycerol, 1, 3-propanediol, panthenol, 1, 2-hexanediol or butanediol.
8. The essence emulsion with anti-aging effect according to claim 6, wherein the preparation raw materials of the essence emulsion with anti-aging effect further comprise, in mass percent: 1-10% of plant source conditioning agent, 0.1-2% of pH regulator, 0.01-0.5% of chelating agent, 0.05-0.5% of antioxidant and 0.1-0.5% of aromatic agent or the combination of at least two.
9. The anti-aging essence emulsion according to claim 8, wherein the plant source conditioner is selected from any one or a combination of at least two of perilla leaf extract, silybum marianum extract and shiitake mushroom extract; preferably, the combination of perilla leaf extract, silybum marianum extract and shiitake mushroom extract.
10. The preparation method of the essence milk with anti-aging effect according to any one of claims 6 to 9, which comprises the following steps:
mixing water and part of polyalcohol in a water phase pot, heating to 60-80deg.C, maintaining the temperature for 10-40min, cooling to 25-40deg.C, and mixing with vitamin A alcohol, glycolic acid, recombinant collagen and chondroitin sulfate to obtain water phase mixture;
mixing the emulsifier, the thickener, the rest polyol, the schizophyllan and the rest raw materials in an emulsifying pot, homogenizing for 5-15min at 30-50 ℃, then mixing with the water phase mixture, and homogenizing for 5-30min at 25-40 ℃ to obtain the essence emulsion with anti-aging effect.
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