CN114983923B - A skin external composition capable of simultaneously resisting aging of internal and external sources - Google Patents
A skin external composition capable of simultaneously resisting aging of internal and external sources Download PDFInfo
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- CN114983923B CN114983923B CN202210819003.2A CN202210819003A CN114983923B CN 114983923 B CN114983923 B CN 114983923B CN 202210819003 A CN202210819003 A CN 202210819003A CN 114983923 B CN114983923 B CN 114983923B
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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
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
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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
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/005—Preparations for sensitive skin
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/04—Plant cells or tissues
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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
- A61K2800/59—Mixtures
- A61K2800/592—Mixtures of compounds complementing their respective functions
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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
- A61K2800/59—Mixtures
- A61K2800/592—Mixtures of compounds complementing their respective functions
- A61K2800/5922—At least two compounds being classified in the same subclass of A61K8/18
-
- 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
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Biotechnology (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Chemical & Material Sciences (AREA)
- Birds (AREA)
- Organic Chemistry (AREA)
- Botany (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Cell Biology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Cosmetics (AREA)
Abstract
The invention discloses a skin external composition capable of simultaneously resisting internal and external aging and a preparation method thereof. The skin external composition comprises the following raw material components: 0.1-30% of anti-aging active, 0.1-15% of humectant, 0.1-15% of emollient, 0.1-12% of emulsifier, 0.1-12% of filler, 0.1-12% of thickener, 0.01-5% of preservative, 0.01-1% of pH regulator, 0.01-1% of daily essence and the balance of deionized water. The main components of the anti-aging active comprise: mirabilis active plant cells, yeast/rice fermentation product filtrate and a lysates of the fermentation products of the two yeasts. The components of the anti-aging active substance in the formula are used simultaneously, so that the anti-aging active substance has synergistic anti-aging effect, can resist endogenous aging, and can repair skin injury caused by exogenous aging.
Description
Technical Field
The invention belongs to the technical field of daily chemicals, and relates to a skin external composition capable of simultaneously resisting internal and external aging and a preparation method thereof.
Background
For skin aging, there are various theories about skin aging, such as genetic theory, radical theory, non-enzymatic glycosylation theory, and the like. Overall, there are two main causes of skin aging, namely endogenous aging and exogenous aging.
Wherein endogenous aging is generally determined by genetic factors, and is an irreversible process of natural aging of skin. With age, in vivo DNA is gradually damaged to cause aging and damage of cells, when mitochondrial dysfunction occurs in cells, ATP supply is reduced, and when cellular activity lacks energy, problems of all physiological activities such as proliferation, differentiation, reduction of collagen, elastin and other proteins of skin cells are caused later, and finally, the skin structure is degenerated to form aged wrinkles. Exogenous aging is mainly caused by environmental factors and life style, such as ultraviolet irradiation, PM2.5, staying up night, eating habit, psychological stress and other problems, wherein the longest problem is that the ultraviolet irradiation, long-term ultraviolet irradiation causes the reduction of collagen fibers and elastic fibers of the skin to cause the aging of the skin, and when the skin loses elasticity and retractive force, the skin becomes more and more loose.
Since skin aging is caused by the combined action of endogenous and exogenous factors, it is generally necessary to pay attention to how to care skin from both aspects in order to delay the progression of aging.
Many related technologies about anti-skin aging, such as chinese patent CN113244156B, disclose an anti-aging composition comprising the following components in weight percent: 0.01 to 0.07 percent of humus extract, 0.001 to 0.05 percent of polyphenol antioxidant, 0.01 to 0.8 percent of mussel extract, 0.05 to 2 percent of sodium hyaluronate, 0.05 to 2 percent of sodium polyglutamate and the balance of deionized water. The anti-aging effect is achieved by the following four aspects: (1) The generation of skin Reactive Oxygen Species (ROS) is reduced by effectively shielding ultraviolet rays and high-energy blue light; (2) effective scavenging of the excess free radicals generated; (3) performing anti-inflammatory repair on damaged skin; (4) The composition can fully exert the functions of the components and produce synergistic effect to realize effective anti-aging effect through reasonable design. Chinese patent CN112932988B discloses an anti-aging composition containing polydeoxyribonucleic acid, comprising honey and polydeoxyribonucleic acid; the disclosed anti-aging composition and skin care product comprising the same enhance skin cell regeneration and anti-aging effects through the synergistic effect of honey and polydeoxyribonucleic acid. Chinese patent CN108392436B discloses a skin care composition for whitening skin and anti-aging, comprising the following components in weight percentage: 10-35% of butanediol, 0.1-5% of glucan, 0.1-50% of lotus leaf flavone, 0.1-5% of nicotinamide and the balance of distilled water. The content of heavy metals and microorganisms is low, so that the whitening cream has remarkable whitening effect, has remarkable anti-aging performance and can remove DPPH free radicals. Chinese patent CN113893203a discloses a skin external composition containing anti-aging liposome and its preparation method. The skin external composition comprises the following raw material components: 0.1-15% of anti-aging efficacy liposome, 1-30% of humectant, 1-22.5% of emollient, 1-21% of emulsifier, 0.1-10% of filler, 1-12.5% of thickener, 0.1-3.5% of preservative, 0.01-1% of pH regulator, 0.01-1% of daily essence and the balance of deionized water. The main components of the anti-aging efficacy liposome comprise: hexapeptide-11, dunaliella salina extract, leucasian willow leaf extract, gynostemma pentaphyllum extract. The preparation method can increase the synthesis of skin collagen, enhance the activity of superoxide dismutase, remove excessive free radicals in skin, relieve peroxidation damage of tissues or cells, strengthen skin barrier, strengthen skin structure and resist skin aging relaxation by simultaneously compounding the components.
Therefore, the related technology of anti-aging skin care products is numerous, and various active ingredients are matched at will, so that how to compound the composition which is effective and can resist the internal and external aging is the key point and the hot point of the research in the cosmetic industry.
Disclosure of Invention
The invention aims to provide a novel skin external composition capable of simultaneously resisting internal and external aging and a preparation method thereof.
The aim of the invention can be achieved by the following technical scheme:
in order to achieve the above object, the present invention adopts the following technical scheme:
the invention firstly provides an anti-aging active composition which comprises the following components: the mass ratio of the mirabilis jalapa active plant cells, the saccharomycetes/meter fermentation product filtrate and the yeast fermentation product lysate is (0.5-5): (1-10): (0.5 to 5), preferably 1: (1-4): 1.
in one embodiment of the invention, the mirabilis active plant cell is prepared by the following preparation method:
1) The mirabilis jalapa leaves are subjected to surface sterilization, crushed and placed in a solid culture medium for culture to obtain leaf callus;
2) Separating leaf callus from Mirabilis jalapa leaves when the leaf callus grows to be 1-3 cm in diameter, performing suspension culture on the leaf callus, placing the leaf callus in continuous fluorescent lamp irradiation, and placing the leaf callus in a shaking table at 24-26 ℃ for subculture to obtain a subculture suspension culture;
3) Separating the subculture suspension, removing tissue mass, dispersing the filtered cell fluid in fresh culture medium, and continuously culturing to obtain the mirabilis jalapa active plant cells.
In one embodiment of the invention, the composition of the solid medium of step 1) is: 2mg/mL NAA, 1mg/mL 6-BA and 1mg/mL 2,4-D were added to the common MS medium.
In one embodiment of the present invention, the medium used for suspension culture in step 2) is: sterilized MS medium, and 2mg/mL NAA, 1mg/mL 6-BA, 1mg/mL 2,4-D were added.
In one embodiment of the invention, the intensity of the continuous fluorescent lamp illumination in step 2) is 4000-6000lux, for example 5000lux may be chosen.
In one embodiment of the present invention, the specific method for performing suspension culture in step 2) is as follows: the callus was removed (the callus was generally soft and loose in structure), gently crushed in a flask, and the MS culture in the flask for sterilization was based on shaking culture on a shaker at 24-26℃at a speed of 120rpm.
In one embodiment of the invention, subculture in step 2) means once every 7d passages.
In one embodiment of the invention, the fresh medium in step 3) is: sterilized MS medium, and 2mg/mL NAA, 1mg/mL 6-BA, 1mg/mL 2,4-D were added.
In one embodiment of the invention, the operation of step 3) is mainly for filtration and expansion culture of cells: the suspension culture obtained at the time of passage was filtered off large tissue masses with a 50-mesh stainless steel screen, and the filtered cell fluid was dispersed in fresh medium for continuous culture.
In one embodiment of the invention, the yeast/rice fermentation product filtrate is a relatively conventional additive in the cosmetic field, and can be obtained by adopting a conventional preparation method or can be directly purchased.
In one embodiment of the invention, the lysate of the fermentation product of Saccharomyces cerevisiae is a Bifidobacterium extract containing metabolites, cytoplasmic components, cell wall components and polysaccharide complexes.
The invention also provides a skin external composition capable of simultaneously resisting the internal and external aging, which comprises the anti-aging active composition and also comprises acceptable auxiliary agents in the skin external product.
In one embodiment of the present invention, the present invention provides a skin external composition capable of simultaneously resisting internal and external aging, which is composed of the following components by weight of the total composition:
0.1-30% of anti-aging active substance, 0.1-15% of humectant, 0.1-15% of emollient, 0.1-12% of emulsifier, 0.1-12% of filler, 0.1-12% of thickener, 0.01-5% of preservative, 0.01-1% of pH regulator, 0.01-1% of daily essence and the balance of deionized water, wherein the anti-aging active substance is the anti-aging active composition.
Preferably, the external skin composition capable of simultaneously resisting the internal and external aging comprises the following components in percentage by weight: 0.1-8% of mirabilis jalapa active plant cells, 0.1-14% of yeast/rice fermentation product filtrate, 0.1-8% of a lysate of a fermentation product of the two-way yeast, 0.1-15% of a humectant, 0.1-15% of an emollient, 0.1-12% of an emulsifying agent, 0.1-12% of a filler, 0.1-12% of a thickening agent, 0.01-5% of a preservative, 0.01-1% of a pH regulator, 0.01-1% of daily essence and the balance of deionized water.
Preferably, the external skin composition capable of simultaneously resisting the internal and external aging comprises the following components in percentage by weight:
0.1-20% of anti-aging active substance, 0.1-10% of humectant, 1-15% of emollient, 1-15% of emulsifier, 0.1-5% of filler, 0.1-8% of thickener, 0.01-3% of preservative, 0.01-0.8% of pH regulator, 0.01-0.8% of daily essence and the balance of deionized water, wherein the anti-aging active substance is the anti-aging active composition.
Preferably, the external skin composition capable of simultaneously resisting the internal and external aging comprises the following components in percentage by weight:
0.1-5% of mirabilis jalapa active plant cells, 0.1-10% of yeast/rice fermentation product filtrate, 0.1-5% of a lysate of a fermentation product of the two-way yeast, 0.1-10% of a humectant, 1-15% of an emollient, 1-15% of an emulsifying agent, 0.1-5% of a filler, 0.1-8% of a thickening agent, 0.01-3% of a preservative, 0.01-0.8% of a pH regulator, 0.01-0.8% of daily essence and the balance of deionized water. Preferably, the external skin composition capable of simultaneously resisting the internal and external aging comprises the following components in percentage by weight:
0.1-15% of an anti-aging active comprising: 0.1-3% of mirabilis active plant cells, 0.1-8% of yeast/rice fermentation product filtrate and 0.1-3% of lysates of fermentation products of saccharomyces cerevisiae;
0.1-8% of humectant, 2-15% of emollient, 2-15% of emulsifier, 0.1-2% of filler, 0.1-5% of thickener, 0.01-2% of preservative, 0.01-0.5% of pH regulator, 0.01-0.4% of daily essence and the balance of deionized water.
Preferably, the skin external composition capable of simultaneously resisting the aging of the internal source and the external source, and the humectant is selected from the following components: one or more of glycerol, butanediol, propylene glycol, pentanediol, phytosterol, dipropylene glycol, hexylene glycol, polyethylene glycol, panthenol, xylitol, sodium hyaluronate, glycerol polyether, methyl glucitol polyether, etc.
The emollient is selected from: one or more of methyl glucose sesquistearate, squalane, beeswax, myristyl alcohol, cetostearyl alcohol, caprylic/capric triglyceride, shea butter, hydrogenated castor oil, jojoba oil, canola oil, olive oil, sunflower seed oil, isohexadecane, and the like;
the emulsifier is selected from: one or more of glycerol stearate, PEG-100 stearate, PEG400 dilaurate, PEG-400 monolaurate, PEG-12 lauryl ether, PEG20 sorbitan monostearate, cetostearyl glucoside, and the like;
the filler is selected from: one or more of polymethylsilsesquioxane, kaolin, silica powder, starch octyl aluminum succinate, talcum powder and the like;
the thickener is selected from: one or more of carbomers, polyethylene, sodium polyacrylate, sodium acrylate/sodium acryloyldimethyl taurate copolymer, acrylic acid (esters) and C10-30 alkanol acrylate cross-linked polymer;
the preservative is selected from the group consisting of: one or more of phenoxyethanol, ethylhexyl glycerol, p-hydroxyacetophenone, sodium benzoate and potassium sorbate;
the pH regulator is one or more of lactic acid, arginine and sodium hydroxide.
Preferably, the preparation method of the skin external composition containing the anti-aging efficacy liposome comprises the following steps:
a method for preparing a skin external composition capable of simultaneously resisting internal and external aging, comprising the following steps:
1) And stirring the humectant and deionized water together, heating to 70-80 ℃, and stirring for 10-30 minutes at a constant temperature to obtain a water phase material.
2) Dissolving the emollient and the emulsifier, stirring and dispersing uniformly, and heating to 70-80 ℃ until the emollient and the emulsifier are completely dissolved to obtain an oil phase material.
3) Slowly adding the oil phase raw material into the water phase material under a homogenizing state of 3000-5000r/min, continuously homogenizing for 5-15min, and then adding the pH regulator, the preservative, the thickener and the filler.
4) Stirring and cooling the mixed solution in the step 3) to 40-50 ℃, adding anti-aging active substances and essence, and homogenizing for 5-10min at 1500-3000 r/min;
5) After homogenization, stirring and cooling to 30-40 ℃ at the speed of 200-300r/min, thus obtaining the skin external composition capable of simultaneously resisting internal and external aging.
The invention also provides the use of the anti-aging active composition capable of simultaneously resisting the internal and external aging, and the anti-aging active composition capable of simultaneously resisting the internal and external aging is used for preparing cosmetics.
In one embodiment of the invention, the cosmetic product comprises a form of face cream, eye cream, lotion, essence, or the like.
The invention discloses a skin external composition capable of simultaneously resisting internal and external aging. The main anti-aging active ingredients of the skin external composition capable of simultaneously resisting the internal and external aging comprise: the optimal composition ratio is found by optimal combination and efficacy test of Mirabilis jalapa active plant cells, yeast/rice fermentation product filtrate and two-way yeast fermentation product lysate, so that the synergistic efficacy of anti-aging repair is obtained.
(1) Mirabilis active plant cell: mirabilis jalapa is a traditional medicine widely used in many parts of the world for treating various diseases. As plant tissue culture techniques develop, innovative mirabilis jalapa active ingredients derived from dedifferentiated plant cell cultures have been developed on the market, with the leaves, stems or roots of which a large number of biologically active compounds of sustainable origin are cultivated. The mirabilis jalapa active plant cells from plant cell culture are verified to enhance the malic acid-aspartic acid shuttle activity, thereby enhancing the energy supply function of mitochondria, so that the mirabilis jalapa active plant cells can show strong anti-aging capability by enhancing the cell metabolic process required by the physiological activity of healthy cells, and the skin becomes tighter and younger.
(2) Yeast/rice fermentation product filtrate: the yeast/rice fermentation product filtrate contains more than 90 nutrition components, the types of the yeast/rice fermentation product filtrate comprise fruit acid, sugar, amino acid, polyamine, nucleic acid and the like, and the most important is that the yeast/rice fermentation product filtrate is rich in alpha-ketoglutarate, can directly participate in tricarboxylic acid circulation to enhance energy metabolism of cells, and provides energy for physiological activities of the cells; simultaneously, mRNA expression of the protein is accurately regulated and controlled, and the cell structure is strengthened from the gene level, so that the aging problem is solved.
(3) Lysates of fermentation products of Saccharomyces cerevisiae: is a Bifidobacterium extract containing metabolites, cytoplasmic components, cell wall components and polysaccharide complex. When the skin is irradiated by ultraviolet rays, the balance between IL-10 and IL-12 is broken, so that the cell DNA is damaged, and the lysate of the fermentation product of the saccharomyces cerevisiae can prevent the release of the immunosuppressant IL-10 and simultaneously prevent the IL-12 from being inhibited, so that the skin can promote the repair mechanism of the cell DNA, help the skin cells and the dermis cells to have the harmful effect on the ultraviolet rays, and keep the skin in a full-activity young appearance.
By means of the technical scheme, the invention has the following beneficial effects:
1) The innovative anti-aging actives of the present invention mainly include: mirabilis active plant cells, yeast/rice fermentation product filtrate, and lysates of yeast fermentation products. Wherein the mirabilis jalapa active plant cells and the yeast/rice fermentation product filtrate can promote the tricarboxylic acid circulation of mitochondria, maintain the energy supply of cells so as to resist endogenous aging, and the yeast fermentation product lysate can help the epidermal cells and dermal cells to resist the damage of external ultraviolet rays, specifically support the protection and repair mechanism of the skin, thereby resisting the premature skin aging.
2) The innovative anti-aging active substance can play a synergistic anti-aging role through effective combination of the components, can resist endogenous aging and can repair skin injury caused by exogenous aging.
Drawings
FIG. 1 is a graph showing changes in the level of malate/aspartate shuttle activity (young cells) in the presence of example 1 at various concentrations;
FIG. 2 shows changes in the levels of malate/aspartate shuttle activity (senescent cells) in the presence of example 1 at various concentrations;
FIG. 3 shows the level of Malondialdehyde (MDA) in the presence of different concentrations of example 1.
Detailed Description
The invention discloses a skin external composition capable of simultaneously resisting internal and external aging. The main anti-aging active ingredients of the skin external composition comprise: the optimal composition ratio is found by optimal combination and efficacy test of Mirabilis jalapa active plant cells, yeast/rice fermentation product filtrate and two-way yeast fermentation product lysate, so that the synergistic efficacy of anti-aging repair is obtained.
The invention is illustrated by the following examples which are given for further illustration of the invention and are not intended to limit the scope of the invention. Some insubstantial modifications and adaptations of the invention by others are within the scope of the invention.
The following percentages (%) are mass percentages unless otherwise indicated. All raw material components of the invention are commercial products.
Examples 1 to 3
An external composition for skin capable of simultaneously resisting internal and external aging is prepared into anti-aging facial cream, and the raw material components and the content are shown in the following table 1.
TABLE 1 raw material composition ratio (unit: mass%) of examples 1-3
A process for preparing a cream composition of the above embodiments 1-3, which is resistant to both internal and external aging, comprising the steps of:
1) And stirring the humectant and deionized water together, heating to 80 ℃, and carrying out heat preservation and stirring for 30 minutes to obtain a water phase material.
2) And dissolving the emollient and the emulsifier, stirring and dispersing uniformly, and heating to 80 ℃ until the emollient and the emulsifier are completely dissolved to obtain an oil phase material.
3) Slowly adding the oil phase raw material into the water phase material in a homogeneous state of 4500r/min, continuously homogenizing for 6min, and then adding the pH regulator, the preservative, the thickener and the filler.
4) Stirring and cooling the mixed solution in the step 3) to 45 ℃, adding the anti-aging active substance and the essence, and homogenizing for 5min at 3000 r/min.
5) After homogenization, stirring at a speed of 200r/min, and cooling to 37 ℃ to obtain the cream composition containing the anti-aging active substances.
Effect verification test example
1. Safety test
Test sample: example 1;
the testing method comprises the following steps: and testing the security by adopting a third party detection mechanism. The selected area is not more than 50mm 2 A suitable plaque assay having a depth of about 1mm is provided wherein about 0.02 to about 0.025ml (g) of the test object is introduced into the plaque assay. The patch tester is attached to the forearm of a subject, the test object is removed after 24 hours, skin reactions are observed after 0.5 hour, 24 hours and 48 hours respectively, and the results are recorded according to the skin reaction grading standard in cosmetic safety technical regulations 2015.
TABLE 2 Patch test results
Wherein, the control group is tested by purified water, and neither stimulation nor anaphylactic reaction occurs.
Remarks: skin reaction grading criterion
Level 0: a negative reaction;
stage 1: suspicious reactions; only weak erythema;
2 stages: weak positive response (erythema response); erythema, infiltration, edema, and possibly papules;
3 stages: strong positive response (herpes response); erythema, infiltration, edema, papules, herpes; the reaction may be beyond the test zone;
4 stages: extremely strong reactions (fusogenic herpetic reactions); obvious erythema, severe infiltration, edema, blepharospermia, and response beyond the test area.
From table 2 above, the prepared cream showed no irritation and was safe. Examples 2 and 3 differ from example 1 only in terms of the amounts used, and are generally safe and non-irritating since no other irritating ingredients are added.
2. Stability test
Test sample: example 1;
the testing method comprises the following steps: internal control standard, the prepared cosmetic compositions were respectively put into the following conditions for stability investigation, CYCLE means that the temperature was cycled from 45 ℃ to-15 ℃ once a day, and the appearance color, viscosity, pH were observed for three months, and the results are shown in table 3.
TABLE 3 stability investigation results
As can be seen from Table 3, the prepared face cream has no significant changes in color, viscosity and pH within three months under high and low temperature or illumination conditions, which indicates that the face cream product of example 1 has better stability.
3. Efficacy evaluation of anti-aging actives
Test sample: anti-aging actives in example 1
The testing method comprises the following steps:
(1) Malic acid/aspartic acid shuttle assay
Using an enzymatic method, a primary culture of human keratinocytes was obtained from 35 and 62 year old donors, the human keratinocytes were seeded onto a semi-permeable polycarbonate membrane and incorporated into a cell culture system and placed in a standard well plate, which placed the culture at the gas-liquid interface (the medium in the well plate was MCDB 153 serum-free medium), and the cells were cultured at the gas-liquid interface for 14 days with medium changes every two days. The epidermal cells thus formed were used for the study since day 14 of culture. The anti-aging active substance in example 1 was co-cultured with human keratinocytes at different concentrations (the concentration means that the anti-aging active substance accounts for the MASs concentration of the culture system, 0.5%, 1%, 2.5% were selected respectively) for 24 hours, and then mitochondria were isolated, and the activity of malate-aspartate shuttle (MAS) was measured. Specifically, 50. Mu.l of the mitochondrial suspension was mixed with 2ml of the assay (300 mM mannitol, 10mM potassium phosphate, 10mM Tris buffer, 10mM potassium chloride, 5mM magnesium chloride, 2mM aspartic acid, 2mM ADP, 0.14mM NADH, 3U/ml Malate Dehydrogenase (MDH), 2U/ml aspartate aminotransferase mitochondria), and pH 7.4. The oxidation of NADH was monitored at a constant temperature of 37℃for 4 minutes at 340 nm. 4mM malic acid and 4mM glutamic acid (final concentration) were added to initiate the malic-aspartic acid shuttle activity. The dose of malic/aspartic shuttle activity was measured at 340nm for 4 minutes.
(2) Assessment of lipid peroxidation degree by Malondialdehyde (MDA)
After the cell suspension was treated after co-culturing the anti-aging actives of example 1 with human keratinocytes at various concentrations (0.5%, 1%, 2.5%) for 24 hours, MDA levels were measured using malondialdehyde content detection kit.
Test results:
(1) Malic acid/aspartic acid shuttle assay: results in young groups of cells (cells from 35 year old donors) the anti-aging actives shown in figure 1 increased malate/aspartate shuttle activity by 20%, 23%, 25% at concentrations of 0.5%, 1%, 2.5%, respectively; results in cells of the aging group (cells from 62 year old donors) the anti-aging actives shown in figure 2 increased malate/aspartate shuttle activity by 18%, 24%, 28% at concentrations of 0.5%, 1%, 2.5%, respectively. The anti-aging active substance can effectively promote the function of mitochondria, thereby realizing the anti-aging function.
(2) Malondialdehyde (MDA) evaluation of lipid peroxidation degree: the final product of lipid peroxidation reaction in organism is malondialdehyde, which can cause cross-linking polymerization of living macromolecules such as protein, nucleic acid and the like, has cytotoxicity, the content of which is an important parameter reflecting the oxidation potential capability of organism, can reflect the lipid peroxidation rate and strength of organism, and can also indirectly reflect the tissue peroxidation damage degree, and the results in figure 3 show that the anti-aging active substance in the embodiment 1 has remarkable protective effect, and the anti-physiological lipid peroxidation is 24%, 30% and 33% respectively at the concentrations of 0.5%, 1.0% and 2.5%.
4. Synergistic verification of topical skin compositions
Test sample: the experimental group was example 1, and the specific composition of the control group is shown in table 4.
TABLE 4 raw material component proportions (unit: mass%) of example 1 and comparative examples 1 to 7
The preparation process of the face cream composition in the above comparative examples 1 to 7 comprises the following steps:
1) And stirring the humectant and deionized water together, heating to 80 ℃, and carrying out heat preservation and stirring for 30 minutes to obtain a water phase material.
2) And dissolving the emollient and the emulsifier, stirring and dispersing uniformly, and heating to 80 ℃ until the emollient and the emulsifier are completely dissolved to obtain an oil phase material.
3) Slowly adding the oil phase raw material into the water phase material in a homogeneous state of 4500r/min, continuously homogenizing for 6min, and then adding the pH regulator, the preservative, the thickener and the filler.
4) Stirring and cooling the mixed solution in the step 3) to 45 ℃, adding the anti-aging active substance and the essence, and homogenizing for 5min at 3000 r/min.
5) After homogenization, stirring at a speed of 200r/min, and cooling to 37 ℃ to obtain the cream composition containing the anti-aging active substances.
The testing method comprises the following steps: after daily face cleaning, a proper amount of test group samples are smeared on one side face randomly, and a control group sample is smeared on the other side. The sunscreen product is applied to the whole face daily in the morning. The results were tested in the following 4 time periods, respectively: before the sample (D0), after 14 days of the sample (D14), after 28 days of the sample (D28), and after 56 days of the sample (D56). The test is performed in a room at 20-22 c with a relative humidity of 40-60% and the subject must now be conditioned for 15 minutes in the same environment.
The detection items are as follows: primos is used for facial image acquisition and skin wrinkle analysis;
the test results are shown in table 5 below.
Table 5 synergistic test set effects
As seen from the results of Table 5, the wrinkle resistance was compared with the comparative results: example 1> comparative examples 5-7> comparative examples 1-3, demonstrate that the selected functional ingredients of mirabilis jalapa active plant cells, yeast/rice fermentation product filtrate, and yeast fermentation product lysate can effectively reduce skin and facial wrinkles by 15.76% when being used in a simultaneous compounding manner, and have better anti-aging effects compared with the use of a single component or two components, which means that the components of the anti-aging active substance in the formula of the invention are used simultaneously, and have synergistic anti-aging effects. In addition, the experimental results of the functional components show that the anti-aging active substance can effectively enhance the mitochondrial activity and provide an energy source for the physiological activities of skin cells. In conclusion, the skin external composition provided by the invention has a remarkable anti-aging effect, can effectively improve the skin aging condition, can delay the arrival of skin aging when the cream prepared by the composition is used for a long time, and can also be seen as younger and more active in the same age.
The previous description of the embodiments is provided to facilitate a person of ordinary skill in the art in order to make and use the present invention. It will be apparent to those skilled in the art that various modifications can be readily made to these embodiments and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above-described embodiments, and those skilled in the art, based on the present disclosure, should make improvements and modifications without departing from the scope of the present invention.
Claims (8)
1. An anti-aging active composition, characterized by comprising the following components: the mass ratio of the mirabilis active plant cells, the saccharomycetes/meter fermentation product filtrate and the yeast fermentation product lysate is 1: (1-4): 1, a step of;
the mirabilis jalapa active plant cell is prepared by the following preparation method:
1) The mirabilis jalapa leaves are subjected to surface sterilization, crushed and placed in a solid culture medium for culture to obtain leaf callus;
2) Separating leaf callus from Mirabilis jalapa leaves when the leaf callus grows to be 1-3 cm in diameter, performing suspension culture on the leaf callus, placing the leaf callus in continuous fluorescent lamp irradiation, and placing the leaf callus in a shaking table at 24-26 ℃ for subculture to obtain a subculture suspension culture;
3) Separating the subculture suspension, removing tissue mass, dispersing the filtered cell fluid in fresh culture medium for continuous culture to obtain the mirabilis jalapa active plant cells;
the solid culture medium in the step 1) comprises the following components: 2mg/mL NAA, 1mg/mL 6-BA and 1mg/mL 2,4-D are added into a common MS culture medium;
the specific method for suspension culture in the step 2) comprises the following steps: taking out the callus, putting the callus into a bottle for crushing, and carrying out shaking culture on a shaking table at 24-26 ℃ for MS culture with sterilization in the bottle, wherein the rotation speed is 120rpm, and the culture medium used for suspension culture in the step 2) is as follows: the sterilized MS culture medium is added with 2mg/mL NAA, 1mg/mL 6-BA and 1mg/mL 2,4-D, the continuous fluorescent lamp irradiation intensity in the step 2) is 4000-6000lux, and the subculture in the step 2) means that every 7 and D passages are carried out;
the fresh medium in step 3) is: sterilized MS medium, and 2mg/mL NAA, 1mg/mL 6-BA, 1mg/mL 2,4-D were added.
2. A skin external composition capable of simultaneously resisting internal and external aging, comprising the anti-aging active composition according to claim 1, and acceptable auxiliary agents in skin external products.
3. The external skin composition capable of simultaneously resisting internal and external aging according to claim 2, which is characterized by comprising the following components in percentage by weight:
0.1-30% of anti-aging active, 0.1-15% of humectant, 0.1-15% of emollient, 0.1-12% of emulsifier, 0.1-12% of filler, 0.1-12% of thickener, 0.01-5% of preservative, 0.01-1% of pH regulator, 0.01-1% of daily essence and the balance of deionized water, wherein the anti-aging active is the anti-aging active composition of claim 1.
4. The external skin composition for simultaneously resisting internal and external aging according to claim 3, wherein the external skin composition for simultaneously resisting internal and external aging comprises the following components in percentage by weight:
0.1-8% of mirabilis jalapa active plant cells, 0.1-14% of yeast/rice fermentation product filtrate, 0.1-8% of a lysate of a fermentation product of the two-way yeast, 0.1-15% of a humectant, 0.1-15% of an emollient, 0.1-12% of an emulsifying agent, 0.1-12% of a filler, 0.1-12% of a thickening agent, 0.01-5% of a preservative, 0.01-1% of a pH regulator, 0.01-1% of daily essence and the balance of deionized water.
5. The external skin composition capable of simultaneously resisting internal and external aging according to claim 2, which is characterized by comprising the following components in percentage by weight:
0.1-20% of anti-aging active, 0.1-10% of humectant, 1-15% of emollient, 1-15% of emulsifier, 0.1-5% of filler, 0.1-8% of thickener, 0.01-3% of preservative, 0.01-0.8% of pH regulator, 0.01-0.8% of daily essence and the balance of deionized water, wherein the anti-aging active is the anti-aging active composition of claim 1 or 2.
6. The external skin composition capable of simultaneously resisting internal and external aging according to claim 5, which is characterized by comprising the following components in percentage by weight:
0.1-5% of mirabilis jalapa active plant cells, 0.1-10% of yeast/rice fermentation product filtrate, 0.1-5% of a lysate of a fermentation product of the two-way yeast, 0.1-10% of a humectant, 1-15% of an emollient, 1-15% of an emulsifying agent, 0.1-5% of a filler, 0.1-8% of a thickening agent, 0.01-3% of a preservative, 0.01-0.8% of a pH regulator, 0.01-0.8% of daily essence and the balance of deionized water.
7. The external skin composition capable of simultaneously resisting internal and external aging according to claim 2, wherein the external skin composition capable of simultaneously resisting internal and external aging comprises the following components in percentage by weight:
0.1-3% of mirabilis jalapa active plant cells, 0.1-8% of yeast/rice fermentation product filtrate, 0.1-3% of a lysate of a fermentation product of the two-way yeast, 0.1-8% of a humectant, 2-15% of an emollient, 2-15% of an emulsifying agent, 0.1-2% of a filler, 0.1-5% of a thickening agent, 0.01-2% of a preservative, 0.01-0.5% of a pH regulator, 0.01-0.4% of daily essence and the balance of deionized water.
8. The external composition for skin which is simultaneously resistant to both internal and external aging according to claim 2, wherein the external composition for skin which is simultaneously resistant to both internal and external aging is a cosmetic.
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