CN117297999A - Composition for improving skin state and application thereof - Google Patents
Composition for improving skin state and application thereof Download PDFInfo
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- CN117297999A CN117297999A CN202310940801.5A CN202310940801A CN117297999A CN 117297999 A CN117297999 A CN 117297999A CN 202310940801 A CN202310940801 A CN 202310940801A CN 117297999 A CN117297999 A CN 117297999A
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- retinol
- ergothioneine
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- 239000000203 mixture Substances 0.000 title claims abstract description 12
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 claims abstract description 194
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- SFRPDSKECHTFQA-ONOWFSFQSA-N [(2e,4e,6e,8e)-3,7-dimethyl-9-(2,6,6-trimethylcyclohexen-1-yl)nona-2,4,6,8-tetraenyl] propanoate Chemical compound CCC(=O)OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C SFRPDSKECHTFQA-ONOWFSFQSA-N 0.000 description 1
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- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 1
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- 229940108325 retinyl palmitate Drugs 0.000 description 1
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Classifications
-
- 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
- A61K8/67—Vitamins
- A61K8/671—Vitamin A; Derivatives thereof, e.g. ester of vitamin A acid, ester of retinol, retinol, retinal
-
- 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
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/494—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with more than one nitrogen as the only hetero atom
- A61K8/4946—Imidazoles or their condensed derivatives, e.g. benzimidazoles
-
- 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
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the 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
-
- 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
- A61K2800/52—Stabilizers
- A61K2800/522—Antioxidants; Radical scavengers
-
- 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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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Cosmetics (AREA)
Abstract
The invention discloses a composition for improving skin condition and application thereof, wherein the addition mass of ergothioneine is 0.01-20%, and the addition mass of retinol is 0.01-20%. The composition for improving skin condition of the present invention can be applied to the preparation of medical and cosmetic products. Ergothioneine can improve stability of retinol, and effectively prevent damage of light and air to retinol. Ergothioneine reduces phototoxic effect of retinol on cells and inflammatory reaction caused by retinol, and compared with retinol alone, the combination of retinol and the ergothioneine promotes proliferation and migration of keratinocytes and basal layer cells better, and shortens cell renewal cycle. Ergothioneine can improve skin irritation side effect caused by retinol, and the combination of the ergothioneine and the retinol has synergistic effect on skin health, and has effects of reducing wrinkles, refining pores, whitening skin, reducing wrinkle, reducing pigmentation, brightening skin, enhancing skin elasticity, promoting collagen and elastin production, resisting inflammation, relieving and enhancing skin barrier.
Description
Technical Field
The invention relates to the technical field of composition preparation, in particular to a composition for improving skin state and application thereof.
Background
Retinol (VA), a fat-soluble vitamin a, is a precursor of retinoic acid (retinoic acid). Retinol or other forms of vitamin a are essential for vision, cellular development, maintenance of skin and mucous membranes, immune function and reproductive development, and are used as an ingredient in skin care products to reduce the effects of wrinkles and other skin aging, including reducing wrinkles, refining pores, whitening and anti-wrinkle, anti-inflammatory and soothing.
The mechanism by which retinol exerts these effects has been demonstrated in studies because it accelerates the renewal of skin cells, causes senescent cells to shed, and promotes the production of new cells. As an all-round skin care active ingredient, retinol is deeply favored by consumers, but the consumers are loved again because of the side effect of skin irritation brought by the retinol. Often consumers will experience varying degrees of skin irritation at the time of initial use, including dryness, irritation, burning, pain, even redness, swelling, peeling, and the like. These side effects are also due to its ability to accelerate cell turnover.
It has been thought by researchers that retinol can update skin cells faster, shed more dead skin cells, but can create a lag time before new healthy cells reach the skin surface. The new skin is exposed to the outside before being prepared and is subject to external influences such as light, air pollutants and the like, with the result that the skin is reddish, discolored and irritated. In addition, retinol itself is poor in stability, is easily denatured and oxidized by exposure to air and light, and may possibly generate substances harmful to the body, and in order to avoid this problem, many of the retinol in the market are modified, such as retinol palmitate, retinol propionate Huang Chunzhi, and the like.
Ergothioneine is a water-soluble small molecular amino acid, widely exists in fungi in nature, exists in animal livers and cannot be synthesized by human bodies, but almost every tissue and organ in the human bodies have the expression of a transporter OCTN-1, and the transporter is also expressed in skin cells. Ergothioneine is a high-efficiency antioxidant, effectively eliminates various free radicals, protects mitochondrial health, delays cell aging, promotes aging cell apoptosis and the like, and is widely used in health care products and cosmetics at present as an antioxidant and anti-aging functional component.
Disclosure of Invention
The invention aims to solve the technical problem that the prior retinol has different skin irritation including side effects of dryness, inflammation, burning sensation, pain, even swelling and peeling and the like when in initial use, and provides a composition for improving skin condition and application thereof.
In order to achieve the above object, the present invention provides a composition for improving skin conditions, wherein ergothioneine is added in an amount of 0.01% -20% by mass, and retinol is added in an amount of 0.01% -20% by mass.
In a preferred embodiment of the present invention, the ergothioneine and the retinol may be added in the same proportion or in different proportions.
In a preferred embodiment of the invention, the retinol is at a cell viability concentration of 70-80%.
The application of the composition for improving the skin state is the application in preparing medical and aesthetic products.
In a preferred embodiment of the present invention, the medical product is one of an aqueous emulsion, an essence, a cream, a mask, a spray, an ampoule, and an oral preparation.
The research shows that ergothioneine can improve the stability of retinol, and effectively prevent the damage of illumination and air to retinol. Experiments show that ergothioneine reduces the toxic effect of retinol on cells, reduces inflammatory reaction caused by retinol, better promotes proliferation and migration of keratinocytes and basal layer cells by combining the retinol and the cell regeneration period by using the ergothioneine and the retinol independently. Thus, it is believed that ergothioneine can ameliorate the skin irritation side effects of retinol, and that the combination of ergothioneine and retinol has a synergistic effect on skin health. Skin health includes reducing wrinkles, refining pores, whitening and anti-wrinkle, reducing pigmentation, lightening skin tone, enhancing skin elasticity, promoting collagen and elastin production, anti-inflammatory and soothing, enhancing skin barrier.
Drawings
FIG. 1 is a bar graph of retinol versus HaCaT cell viability according to various concentrations.
FIG. 2 is a bar graph of ergothioneine improving the toxic effects of retinol.
FIGS. 3a to 3c are graphs showing the relative expression of genes associated with retinol-induced inflammation according to various concentrations of ergothioneine.
FIG. 4 is a schematic representation of cell scratch images according to example 4 at various time points.
FIG. 5 is a schematic representation of cell scratch images according to example 5 at various time points.
FIG. 6 is a schematic representation of the growth cycle of HaCaT cells according to the presence of retinol in example 6.
Detailed Description
The present invention will be described in detail with reference to specific examples.
EXAMPLE 1 retinol cytotoxicity assay
Human immortalized keratinocytes (HaCaT) in logarithmic growth phase were selected at 1.0X10 per well 4 Inoculated in 96-well cell culture plates (100. Mu.L per well) placed in CO 2 Culturing in an incubator (37 ℃, 5%) for 24 hours. The medium in the 96-well plate was removed, 200. Mu.L of fresh medium containing retinol at different concentrations was added as a dosing group, 200. Mu.L of fresh medium was added as a blank, and after further culturing in an incubator for 24 hours, 100. Mu.L of 10% CCK-8 working solution was replaced, and after 1 hour incubation at 37℃the OD value of each well was measured at 450 nm. Cell viability= (sample group OD/blank group OD value) ×100%.
From a bar graph of retinol versus HaCaT cell viability at different concentrations (see fig. 1), it can be seen that retinol as a dosing group decreases HaCaT cell viability with increasing concentration, i.e. exhibits toxic effects above a certain concentration, with stimulatory effects. We therefore selected a concentration around 75% of cell viability as the subsequent retinol-stimulated dosing concentration to complete the following test and explore the improving effect of ergothioneine on retinol in the present invention.
EXAMPLE 2 ergothioneine improves retinol toxicity
The toxic effect of ergothioneine on HaCaT cells was first determined with only varying concentrations, as in example 1. Except that retinol concentration with cell activity of 70-80% in example 1 was selected as positive control group, three safe concentrations of ergothioneine without significant toxicity were selected as experimental group, the ergothioneine and retinol were compounded and used as fresh dosing medium, and after 24 hours of incubation, 100 μl of 10% cck-8 working solution was replaced, after 1 hour of incubation, OD value of each well was measured at 450nm wavelength, and cell viability was calculated. The improvement effect of ergothioneine on retinol toxicity was determined.
From the results in the bar graph (see FIG. 2), it was shown that ergothioneine alone at a concentration of 0.1mM-10mM has no toxic effect on HaCaT cells. And the ergothioneine at the concentration is correspondingly compounded with the retinol, so that the result shows that the ergothioneine obviously reduces the toxic effect generated by retinol stimulation along with the increase of the concentration, and effectively improves the side effect generated by retinol.
Example 3 ergothioneine improves retinol inflammatory action
HaCaT cells in logarithmic growth phase were selected at 6.0X10 per well 5 Inoculating into 6-well plate with liquid amount of 1mL per well, placing at 37deg.C, and 5% CO 2 Incubation for 24h, the normal control group was replaced with fresh medium, the model control group was replaced with fresh medium containing retinol as defined in example 1, the sample group was replaced with fresh medium containing three safe concentrations of ergothioneine in combination with retinol, at 37 ℃,5% co 2 Incubation was carried out for 24h. Collecting cells: extracting total RNA of each experimental group, synthesizing cDNA, and detecting gene expression of beta-actin and target genes by q-PCR. The relative expression amounts of RNA of TRPV1, IL-1 beta and NF-kappa B genes are calculated by using beta-actin as an internal reference of gene expression.
According to the relative expression patterns of the genes related to the inflammation induced by the retinol with different concentrations of the ergothioneine, referring to fig. 3, it can be seen from fig. 3 that the ergothioneine significantly reduces the expression of the TRPV1, IL-1 beta and NF- κb genes generated by the retinol effect with the increase of the concentration, which indicates that the ergothioneine can effectively relieve the inflammation caused by the retinol on cells and repair the skin barrier.
Example 4 ergothioneine in combination with retinol promotes proliferation and migration of keratinocytes
Selecting HaCaT in logarithmic growth phase, and selecting 5.0X10 of HaCaT per hole 5 Inoculated in 12-well cell culture plates (1 mL per well) placed in CO 2 Culturing in incubator (37 ℃, 5%) for 24h. After the alignment line is marked at the bottom of the culture plate, straight artificial scratches are carried out on each hole of cells by using a 1mL suction head, the culture medium in a 6-hole plate is removed, the cells are washed 3 times by using precooled PBS to wash away cell residues, 1mL of fresh serum-free culture medium containing safe concentrations of independent ergothioneine, independent retinol and ergothioneine compounded with retinol is added as a dosing group, 1mL of fresh culture solution is added as a blank control, EGF growth factors are used as a control group, after the cells are continuously cultured in an incubator for 12 hours, 24 hours and 48 hours respectively, the original scratches are photographed, the comparison analysis is carried out with the photographs at the time of 0 hour, and the cell scratch area analysis HaCaT cell migration capacity is calculated by Image J Image analysis software. After 48h photographing, adding CCK-8 working solution to measure the activity of each group of cells, and calculating the proliferation condition of HaCaT cells
Referring to fig. 4, from the images of cell scratches at different time points, it can be seen that the migration ability of HaCaT cells in the presence of ergothioneine and retinol is significantly improved, the scratch area gradually decreases with time, and finally returns to almost the front of the scratch. Meanwhile, the migration capacity of the combination of ergothioneine and retinol is weaker than that of the retinol, which indicates that the combination of ergothioneine and retinol can play a role in regulating and controlling the migration of skin keratinocytes. In addition, according to the cell viability after 48 hours, both ergothioneine and retinol have a certain proliferation effect on HaCaT cells, and these results indicate that ergothioneine and retinol and their complexes may have the efficacy of rapidly repairing skin barriers and recovering wounds after surgery.
Example 5 ergothioneine in combination with retinol promotes proliferation and migration of basal layer cells
Selecting Human Skin Fibroblast (HSF) in logarithmic growth phase, 1.0X10 per well 5 Inoculated in 12-well cell culture plates (1 mL per well) placed in CO 2 Culturing in an incubator (37 ℃, 5%) for 24 hours. After alignment was established at the bottom of the plate, cells from each well were streaked straight with a 1mL tip, medium from the 6-well plate was removed, cell debris was washed 3 times with pre-chilled PBS, 1mL fresh serum-free medium containing safe concentrations of ergothioneine alone, retinol alone, and ergothioneine in combination with retinol was added as a dosing group, and blank control was added1mL of fresh culture solution, the control group uses EGF growth factors, after the EGF growth factors are continuously cultured in an incubator for 12 hours, 24 hours and 48 hours respectively, the original scratch position is photographed, the comparison analysis is carried out on the original scratch position and the photograph at the time of 0 hour scratch, and the cell scratch area analysis HSF cell migration capacity is calculated through Image J Image analysis software. And (3) after photographing at 48h, adding CCK-8 working solution to measure the activity of each group of cells, and calculating the proliferation condition of the fibroblasts.
Referring to fig. 5, from the images of cell scratches at different time points, it can be seen that the migration ability of HSF cells in the presence of ergothioneine and retinol is significantly improved, the scratch area gradually decreases with time, and finally returns to almost the front of the scratch. Meanwhile, the migration capacity of the combination of the ergothioneine and the retinol is higher than that of the retinol, which indicates that the combination of the ergothioneine and the retinol can play a role in regulating and controlling the migration of the dermis layer into fibers. In addition, according to the cell viability after 48 hours, both ergothioneine and retinol have a certain proliferation effect on HSF cells, and these results indicate that ergothioneine can significantly promote proliferation and migration of dermal fibroblasts, and this effect is also shown when compared with retinol alone in Huang Chunfu, which indicates that ergothioneine can improve the skin sensitivity problem that the proliferation and differentiation speed of dermal cells is lower than that of keratinocytes in the presence of retinol, promote metabolism of whole-layer skin cells, and improve the skin toxicity effect of retinol.
Example 6 ergothioneine in combination with retinol regulates keratinocyte growth cycle
HaCaT cells in logarithmic growth phase were selected at 6.0X10 per well 5 Inoculated in 6-well cell culture plates (1 mL per well) placed in CO 2 Culturing in an incubator (37 ℃, 5%) for 24 hours. Removing culture medium in 6-well plate, washing 3 times with pre-cooled PBS to wash cell residue, adding 1mL fresh culture medium containing ergothioneine and retinol at safe concentration as administration group, adding 1mL fresh culture medium according to safe retinol concentration as positive control group in example 1, continuously culturing in incubator for 24 times, digesting each well cell with pancreatin, detecting cell growth cycle change by cell viability analyzer, analyzing wheatChanges in HaCaT cell growth and proliferation cycle due to both kerathionine and retinol.
Referring to FIG. 6, it can be seen from the growth cycle curve of HaCaT cells in the presence of retinol that when ergothioneine is used with view Huang Chunfu, the excessively fast cell production cycle caused by retinol is effectively controlled, indicating that ergothioneine regulates the metabolism of cells caused by retinol, and prevents skin problems caused by excessive skin renewal.
EXAMPLE 7 ergothioneine enhances retinol stability
The change in ergothioneine and retinol levels was monitored by High Performance Liquid Chromatography (HPLC) as follows:
the method for detecting the content of the ergothioneine by using an HPLC method comprises the following steps:
liquid phase: thermo Fisher
Chromatographic column: c18 reverse chromatography column
Flow rate: 1.0mL/min
Column temperature: 35 DEG C
Mobile phase: 95% methanol-water
And (3) detection: 325 nm/full wavelength
Sample injection amount: 10 mu L
Through the combination of ergothioneine and retinol with different concentrations, 30% ethanol is added for different time points: the content changes of ergothioneine and retinol were measured by HPLC from 1day, 3day, and 7day solutions. The effect of ergothioneine on retinol decomposition was analyzed.
The results show that the retinol content decreases with time, indicating that retinol readily decomposes in 30% aqueous ethanol. When the ergothioneine is matched with the vitamin Huang Chunfu, the decomposition effect is obviously reduced, which proves that the ergothioneine can obviously inhibit the degradation speed of the retinol and improve the stability of the retinol.
Example 8 ergothioneine improves skin discomfort due to retinol
20 women aged 25-35 years had no related skin care products with retinol or EGT used for a period of one year. The left and right faces were treated with 1% retinol essence and 1% retinol &1% egt essence, respectively, once a day for 7 days. At Day 0 and Day 30, the subject would receive a questionnaire, fill out facial skin sensations, and the researcher would record the subject's skin status by way of observation and interrogation. The results were as follows:
8.1 facial analysis and personal statistics of 1% Retinol extract smeared
| Day 0 number of people | Day 7 number of people | Proportion of persons with skin discomfort | |
| Red and swollen | 0 | 4 | 20％ |
| Burning sensation | 0 | 8 | 40％ |
| Tingling sensation | 0 | 6 | 30％ |
| Peeling off | 0 | 5 | 25％ |
| Drying | 0 | 13 | 65％ |
8.2 facial Condition analysis and personal statistics with 1% Retinol &1% EGT essence applied
| Day 0 number of people | Day 7 number of people | Proportion of persons with skin discomfort | |
| Red and swollen | 0 | 1 | 5％ |
| Burning sensation | 0 | 3 | 15％ |
| Tingling sensation | 0 | 5 | 25％ |
| Peeling off | 0 | 3 | 15％ |
| Drying | 0 | 5 | 25％ |
From the analysis of the results, the retinol essence added with EGT can obviously relieve the skin irritation phenomena such as red swelling and the like.
Claims (4)
1. A composition for improving skin condition, characterized in that ergothioneine is added in an amount of 0.01-20% by mass and retinol is added in an amount of 0.01-20% by mass.
2. A composition for improving skin conditions according to claim 1, wherein said ergothioneine and said retinol are added in the same or different proportions.
3. Use of a composition for improving the condition of the skin according to any one of claims 1 to 3, characterized by the use in the preparation of a medical product.
4. The use according to claim 4, wherein the medical product is one of an aqueous emulsion, an essence, a cream, a mask, a spray, an ampoule, and an oral preparation.
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| CN202310940801.5A CN117297999A (en) | 2023-07-28 | 2023-07-28 | Composition for improving skin state and application thereof |
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| CN202310940801.5A CN117297999A (en) | 2023-07-28 | 2023-07-28 | Composition for improving skin state and application thereof |
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