Skin anti-carbonylation composition, preparation method and skin care product containing skin anti-carbonylation composition
By co-culturing cystellae and chlorophyllium trigonoplasma on adenosine culture medium, skin anti-carbonylation compositions are prepared for use in skin care products, and the problem of dull skin yellowing caused by accumulation of carbonylation compositions in the prior art is solved. , achieving stronger antioxidant and cell metabolism activation effects.
Patent Information
- Application Number
- CN202411256900.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-09
- Publication Date
- 2024-12-13
AI Technical Summary
The prior art is not effective in reducing the formation and accumulation of skin carbonylated compositions and cannot effectively improve the problem of dull yellowing and rough skin.
By co-culturing cysteiniae and chlorophyllium trigonoplasma on adenosine-containing culture medium, high-speed homogenization and filtration, skin anti-carbonylation compositions were prepared and used in skin care products to activate cell metabolism and proteasome expression, degrading carbonylated proteins.
Significantly reduces the production and accumulation of carbonylated proteins, improves dull yellowing and rough skin problems, enhances cell viability and proteasome expression, and provides stronger anti-carbonylation effects.
Smart Images
Figure CN119112718A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of skin care products, and in particular relates to a skin anti-carbonylation composition and a preparation method, and skin care products containing the composition. Background Art
[0002] Dark yellow skin is one of the skin problems that bothers Chinese women and is also a hot research and development topic for the cosmetics industry. There are two main factors that lead to dark yellow skin: one is the formation and distribution of melanin, and the other is protein oxidation. To solve the problem of dark yellow skin, in addition to regulating the production and metabolism of melanin, we must also focus on how to avoid the production of oxidized proteins and how to accelerate the metabolism of oxidized proteins. Oxidized proteins are divided into two categories in the skin, one is glycated protein - advanced glycation end products (AGEs), and the other is carbonylated protein - carbonylated protein (CPs), and the latter causes much more yellowing of the skin than the former. Therefore, carbonylated protein is an important target for solving dark yellow skin.
[0003] CPs are formed by the reaction of amino groups in proteins with active aldehyde compounds generated by lipid peroxidation caused by ROS. They are early markers of oxidative stress and accumulate with age. They are difficult to degrade, which can overload the cell's repair and degradation system, exacerbate the accumulation of CPs, and form a vicious cycle, ultimately causing the skin to become yellow and dark, and its water-holding capacity to decrease, leading to a series of problems such as enlarged pores. The carbonylation process of proteins is an irreversible post-translational modification that is closely related to many skin problems and aging.
[0004] Most of the existing active ingredients for solving the problem of skin carbonylation reduce the generation of CPs through antioxidant pathways, but have no effect on the accumulated CPs. In addition, they cannot activate the degradation system of aging cells with excessive accumulation of CPs, resulting in poor anti-carbonylation effect and less than ideal effect on improving skin dullness. Summary of the invention
[0005] The purpose of the present invention is to solve at least one problem in the prior art and to provide a skin anti-carbonylation composition and a preparation method, and a skin care product containing the composition.
[0006] To achieve the above object, the present invention provides a method for preparing a skin anti-carbonylation composition, comprising the following steps:
[0007] Inoculate marine coccolithophore and triangular brown finger algae on f / 2 culture medium containing adenosine, place in a constant temperature photoreactor for cultivation after inoculation, and supplement 0.01% to 0.05% adenosine at regular intervals;
[0008] After the marine coccolithophores and triangular brown finger algae are cultured to a stable growth period, the algae cells are collected by filtration and freeze-dried to obtain algae powder;
[0009] The algae powder is dispersed in an organic solution, subjected to high-speed homogenization treatment, 0.5% to 1% adenosine is added, and the supernatant is filtered to obtain a skin anti-carbonylation composition.
[0010] Preferably, during the process of culturing the marine coccolithophore and the triangular brown finger algae in the constant temperature photoreactor, the marine coccolithophore and the triangular brown finger algae are irradiated with a white LED light source, and the light intensity is adjusted to 50 μE / (m 2 ·s)~100μE / (m 2 ·s), the light cycle was 14L:10D, the culture temperature was 15℃~20℃, and 0.01%~0.05% adenosine was supplemented every 24h.
[0011] Preferably, the algal cells are collected by filtration using a disc filter.
[0012] Preferably, the density of inoculating Coccolithophore and Pseudoalterium triangularis on the f / 2 culture medium is controlled at 10,000 cells / ml to 150,000 cells / ml.
[0013] Preferably, the organic solution is a 20% to 50% propylene glycol aqueous solution or a butylene glycol aqueous solution, and the high-speed homogenization treatment is carried out at a rotation speed of 5000 rpm to 5000 rpm.
[0014] Preferably, the filtering and supernatant extraction treatment is performed through a filter membrane with a pore size of 0.15 μm to 0.25 μm.
[0015] The present invention also provides a skin anti-carbonylation composition prepared according to the preparation method of the skin anti-carbonylation composition.
[0016] The present invention also provides a skin care product, which contains 0.5% to 5% of the skin anti-carbonylation composition.
[0017] Preferably, the skin care product further comprises the following components: 3% to 10% moisturizer, 0.3% to 3% thickener, 3% to 30% emollient, 1% to 8% emulsifier, 0.05% to 8% fragrance agent, 0.1% to 2% preservative and the balance water, wherein the moisturizer comprises one or more of glycerin, butylene glycol, propylene glycol, glyceryl polyether-26, sodium hyaluronate and allantoin, and the emollient comprises isononyl isononanoate, polydimethylsiloxane, caprylic / capric triglyceride, vegetable wax, paraffin, shea butter, cocoa seed butter, tocopherol and tocopherol acetate , cetearyl alcohol, the thickener comprises one or more of carbomer, sodium carbomer, xanthan gum, polyacrylate polymers, and grafted starch, the emulsifier comprises one or more of glyceryl stearate, PEG-100 stearate, polyglycerol emulsifier, hydrogenated lecithin, and tri(laureth-4) phosphate, the preservative comprises one or more of caprylhydroxamic acid, p-hydroxyacetophenone, caprylyl glycol, ethylhexylglycerin, and phenoxyethanol, and the flavoring agent comprises one or more of plant floral water, plant extracts, and plant essential oils.
[0018] Preferably, the skin care product is an essence, a cream or an emulsion.
[0019] Introduction to the efficacy of each ingredient: Adenosine is an endogenous purine nucleoside, a small molecule produced by the denucleation of adenosine triphosphate (ATP) and adenine nucleotides. It can directly replenish cellular energy and produce a variety of physiological effects by activating specific G protein-coupled receptors, activating cellular autophagy levels and maintaining normal cellular energy metabolism; Pseudomonas triangularis and Coccolithophore are two marine microalgae with strong adaptability in the ocean. While synthesizing macromolecular active chemicals such as proteins and polysaccharides during their growth, marine microalgae also synthesize some highly active organic small molecules, especially small molecule secondary metabolites in microalgae, which have important application potential in the field of functional food and drug development.
[0020] The beneficial effects of the present invention are as follows: the present invention inoculates marine coccolithophore and triangular brown finger algae on a culture medium for co-cultivation, and regularly supplements adenosine to activate the metabolism of algal cells while enriching adenosine in the subsequent filtrate, thereby improving the dissolution rate and bioavailability of adenosine, so that adenosine can also activate skin cells when applied to the skin; the skin anti-carbonylation composition obtained by mixed culture proliferation of marine coccolithophore, triangular brown finger algae and adenosine and wall-breaking purification can reduce the generation of CPs by anti-oxidation and increasing the expression of heat shock proteins, increase the expression of proteasomes to degrade the generated CPs, and activate senescent cells to successfully complete the above two metabolic processes, thereby solving the problem of dark yellow and rough skin caused by CPs accumulation from three dimensions of reducing the generation of CPs, degrading the generated CPs and activating the metabolism of cells with CPs accumulation, and compared with a single active ingredient such as triangular brown finger algae, the pathway is more complete and the anti-carbonylation effect is stronger.
[0021] The features and advantages of the present invention will be described in detail through embodiments in conjunction with the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS
[0022] Figure 1 It is a schematic diagram comparing the effects of different components and concentrations of active substances on cell viability according to the embodiments of the present invention.
[0023] Figure 2 It is a schematic diagram of the effect of the skin anti-carbonylation composition according to an embodiment of the present invention on the expression of proteasome-related genes. DETAILED DESCRIPTION
[0024] In order to make the purpose, technical solution and advantages of the embodiments of the present application clearer, the technical solution in the embodiments of the present application will be clearly and completely described below in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, not all of the embodiments.
[0025] Example 1
[0026] This embodiment provides a method for preparing a skin anti-carbonylation composition, comprising the following steps:
[0027] S01. Inoculate marine coccolithophores and triangular brown finger algae on f / 2 culture medium containing 0.1% adenosine, and control the inoculation density at 30,000 cells / ml. After inoculation, place the cells in a constant temperature photoreactor for cultivation so that the marine coccolithophores and triangular brown finger algae are activated and expanded to the logarithmic growth phase. During the cultivation process, white LED light source is used to irradiate the marine coccolithophores and triangular brown finger algae, and the light intensity is adjusted to 60μE / (m 2 ·s), the photoperiod was 14L:10D, the culture temperature was 16°C, and 0.03% adenosine was supplemented every 24 h;
[0028] S02. After the marine coccolithophore and the triangular brown finger algae are cultured to a stable growth period, they are filtered using a disc filter, and the algal cells on the 2.0 μm filter membrane on the disc filter are collected and freeze-dried to obtain algal powder;
[0029] S03. Disperse the algae powder in a 30% propylene glycol aqueous solution, perform high-speed homogenization at a rotation speed of 10,000 rpm, then add 0.5% adenosine, filter with a filter membrane with a pore size of 0.2 μm, take the supernatant, and obtain a skin anti-carbonylation composition.
[0030] The present embodiment also provides a skin care product, which is an essence, and the essence is composed of the following components in percentage by mass: 3% of the skin anti-carbonylation composition of the present embodiment, 5% of lavender flower water, 0.05% of xanthan gum, 0.15% of carbomer sodium, 5% of glycerol, 5% of butylene glycol, 2% of tri(laureth-4) phosphate, 0.5% of hydrogenated lecithin, 2% of polydimethylsiloxane, 5% of caprylic / capric triglyceride, 0.5% of parahydroxyacetophenone, and 71.8% of water.
[0031] The essence is prepared according to the following preparation method: water, xanthan gum, sodium carbomer, glycerin and butylene glycol are weighed and mixed as phase A according to the formula ratio of the essence, tri(laureth-4) phosphate, hydrogenated lecithin, polydimethylsiloxane and caprylic / capric triglyceride are weighed and mixed as phase B according to the formula ratio of the essence, and p-hydroxyacetophenone and lavender flower water are weighed and mixed as phase C according to the formula ratio of the essence; phase A and phase B are heated to 85°C respectively, and after keeping the heat for 30 minutes, phase B is slowly added to phase A, emulsified and homogenized at a rotation speed of 3000rpm for 5 minutes, stirred and cooled to 45°C, and the skin anti-carbonylation composition and phase C in the formula ratio of the essence are added, and stirred evenly to obtain the essence.
[0032] Example 2
[0033] This embodiment provides a method for preparing a skin anti-carbonylation composition, comprising the following steps:
[0034] S01. Inoculate marine coccolithophores and triangular brown finger algae on f / 2 culture medium containing 0.2% adenosine, and control the inoculation density at 50,000 cells / ml. After inoculation, place the culture in a constant temperature photoreactor to activate and expand the marine coccolithophores and triangular brown finger algae to the logarithmic growth phase. During the culture process, use a white LED light source to irradiate the marine coccolithophores and triangular brown finger algae, and adjust the light intensity to 100μE / (m 2 ·s), the photoperiod was 14L:10D, the culture temperature was 20°C, and 0.05% adenosine was supplemented every 24 h;
[0035] S02. After the marine coccolithophore and the triangular brown finger algae are cultured to a stable growth period, they are filtered using a disc filter, and the algal cells on the 2.2 μm filter membrane on the disc filter are collected and freeze-dried to obtain algal powder;
[0036] S03. The algae powder is dispersed in a 40% propylene glycol aqueous solution, and high-speed homogenization is performed at a rotation speed of 15,000 rpm. Then, 1% adenosine is added, and then filtered with a filter membrane with a pore size of 0.18 μm, and the supernatant is taken to obtain a skin anti-carbonylation composition.
[0037] Example 3
[0038] This embodiment provides a method for preparing a skin anti-carbonylation composition, comprising the following steps:
[0039] S01. Inoculate marine coccolithophores and triangular brown finger algae on f / 2 culture medium containing 0.15% adenosine, and control the inoculation density at 40,000 cells / ml. After inoculation, place the cells in a constant temperature photoreactor for cultivation so that the marine coccolithophores and triangular brown finger algae are activated and expanded to the logarithmic growth phase. During the cultivation process, white LED light source is used to irradiate the marine coccolithophores and triangular brown finger algae, and the light intensity is adjusted to 50μE / (m 2 ·s), the photoperiod was 14L:10D, the culture temperature was 18°C, and 0.01% adenosine was supplemented every 24 h;
[0040] S02. After the marine coccolithophore and the triangular brown finger algae are cultured to a stable growth period, they are filtered using a disc filter, and the algal cells on the 2.0 μm filter membrane on the disc filter are collected and freeze-dried to obtain algal powder;
[0041] S03. The algae powder is dispersed in a 40% butanediol aqueous solution, and high-speed homogenization is performed at a rotation speed of 12000 rpm. Then, 0.7% adenosine is added, and then filtered with a filter membrane with a pore size of 0.15 μm, and the supernatant is taken to obtain a skin anti-carbonylation composition.
[0042] The present embodiment also provides a skin care product, which is a facial cream, and the facial cream is composed of the following components in percentage by mass: 5% of the skin anti-carbonylation composition of the present embodiment, 0.05% of sodium hyaluronate, 0.2% of arginine, 0.3% of sodium polyacrylate grafted starch, 0.2% of carbomer, 5% of glyceryl polyether-26, 5% of butylene glycol, 1% of glyceryl stearate, 1% of PEG-100 stearate, 5% of avocado butter, 2% of cetearyl alcohol, 5% of polydimethylsiloxane, 5% of caprylic / capric triglyceride, 0.5% of parahydroxyacetophenone, 0.05% of sandalwood essential oil, and 64.7% of water.
[0043] The face cream is prepared according to the following preparation method: water, sodium hyaluronate, arginine, sodium polyacrylate grafted starch, carbomer, glyceryl polyether-26, and butylene glycol are weighed and mixed as phase A according to the formula ratio of the above face cream; glyceryl stearate, PEG-100 stearate, avocado tree butter, cetearyl alcohol, polydimethylsiloxane, caprylic / capric triglyceride are weighed and mixed as phase B according to the formula ratio of the above essence; p-hydroxyacetophenone and sandalwood essential oil are weighed and mixed as phase C according to the formula ratio of the above essence; phase A and phase B are heated to 85°C respectively, and after keeping the heat for 30 minutes, phase B is slowly added to phase A, emulsified and homogenized at a rotation speed of 5000rpm for 5 minutes, stirred and cooled to 45°C, and the skin anti-carbonylation composition and phase C in the formula ratio of the above essence are added, and stirred evenly to obtain the face cream.
[0044] 1. Experiment on the anti-carbonylation effect of different active ingredients
[0045] Six groups of the same EMEM culture medium were taken, and fibroblasts were cultured under the same conditions on 96-well plates of each culture medium. One group of fibroblasts was not treated, and the other five groups of fibroblasts were treated with equal amounts of 0.1% skin anti-carbonylation composition, 0.5% skin anti-carbonylation composition, 0.1% adenosine, 0.1% triangular brown finger algae (abbreviated as brown finger algae), and 0.1% marine coccolithophore (abbreviated as coccolithophore) for 24 hours, and then 20 μM H2O2 solution was added for treatment for 90 minutes to simulate the cell oxidative stress process. After the treatment, the cells were kept warm at 37°C in MTT solution for 3 hours, and then the MTT solution was removed, 200 μl DMSO was added to each well to lyse the cells, and the absorbance of the supernatant at 570 nm was read using a spectrophotometer after centrifugation. By comparing the samples not treated with H2O2, the protective effect of the complex solution on the oxidative stress process of the cells can be judged. The specific test results are as follows: Figure 1 shown.
[0046] Depend on Figure 1 It can be seen that under the same concentration conditions, the cell viability after adding the skin anti-carbonylation composition is better than the cell viability after adding a single ingredient (adenosine, triangular finger algae, and marine coccolithophore); the cell viability after adding 0.5% of the skin anti-carbonylation composition is better than the cell viability after adding 0.1% of the skin anti-carbonylation composition.
[0047] This indicates that the skin anti-carbonylation composition has a more significant protective effect on cells subjected to oxidative stress, can effectively reduce oxidative stress damage to cells, enhance cell viability, reduce the production of carbonylated proteins, and activate cell metabolism of CPs accumulation. The anti-carbonylation effect of the skin anti-carbonylation composition at a specific concentration, such as 0.5%, is even better.
[0048] 2. Experiment on the effect of skin anti-carbonylation composition on cell proteasome expression
[0049] Fibroblasts were cultured in 96-well plates in EMEM medium and treated with 0.1% complex solution for 24 h. After mechanical disruption of the cells, the quantity and quality of specific RNA synthesis were evaluated by capillary electrophoresis. The specific test results are shown in Figure 2 shown.
[0050] Depend on Figure 2 It can be seen that the skin anti-carbonylation composition significantly increased the expression of several genes related to cellular proteasome synthesis (PSMB8, PSMB9, PSMB10, PSM1 and PSME2), indicating that the skin anti-carbonylation composition has a promoting effect on the proteasome expression of fibroblasts, thereby improving the cell's ability to degrade accumulated carbonylated proteins.
[0051] The above embodiments are intended to illustrate the present invention, not to limit the present invention. Any solution that is a simple transformation of the present invention belongs to the protection scope of the present invention.
Claims
1. A method for preparing a skin anti-carbonylation composition, characterized in that: It includes the following steps: Inoculate marine coccolithophore and triangular brown finger algae on f / 2 culture medium containing adenosine, place in a constant temperature photoreactor for cultivation after inoculation, and supplement 0.01% to 0.05% adenosine at regular intervals; After the marine coccolithophores and triangular brown finger algae are cultured to a stable growth period, the algae cells are collected by filtration and freeze-dried to obtain algae powder; The algae powder is dispersed in an organic solution, subjected to high-speed homogenization treatment, 0.5% to 1% adenosine is added, and the supernatant is filtered to obtain a skin anti-carbonylation composition.
2. The method for preparing the skin anti-carbonylation composition according to claim 1, characterized in that: During the process of culturing marine coccolithophores and triangular brown finger algae in the constant temperature photoreactor, a white LED light source is used to irradiate the marine coccolithophores and triangular brown finger algae, and the light intensity is adjusted to 50 μE / (m 2 ·s)~100μE / (m 2 ·s), the light cycle was 14L:10D, the culture temperature was 15℃~20℃, and 0.01%~0.05% adenosine was supplemented every 24h.
3. The method for preparing the skin anti-carbonylation composition according to claim 1, characterized in that: The algal cells are collected by filtration using a disc filter.
4. The method for preparing the skin anti-carbonylation composition according to claim 1, characterized in that: The density of inoculating ocean coccolithophore and triangular brown finger algae on the f / 2 culture medium is controlled to be 10000 cells / ml to 50000 cells / ml.
5. The method for preparing the skin anti-carbonylation composition according to claim 1, characterized in that: The organic solution is a 20% to 50% propylene glycol aqueous solution or a butylene glycol aqueous solution, and the high-speed homogenization treatment is carried out at a rotation speed of 5000 rpm to 15000 rpm.
6. The method for preparing the skin anti-carbonylation composition according to claim 1, characterized in that: The filtering and supernatant treatment is carried out through a filter membrane with a pore size of 0.15 μm to 0.25 μm.
7. A skin anti-carbonylation composition prepared according to the method for preparing a skin anti-carbonylation composition according to any one of claims 1 to 6.
8. A skin care product, characterized in that: The skin care product comprises 0.5% to 5% of the skin anti-carbonylation composition according to claim 7.
9. The skin care product according to claim 8, characterized in that The skin care product also includes the following components: 3% to 10% moisturizer, 0.3% to 3% thickener, 3% to 30% emollient, 1% to 8% emulsifier, 0.05% to 8% fragrance, 0.1% to 2% preservative and the balance water, the moisturizer includes one or more of glycerin, butylene glycol, propylene glycol, glyceryl polyether-26, sodium hyaluronate, and allantoin, the emollient includes isononyl isononanoate, polydimethylsiloxane, caprylic / capric triglyceride, vegetable wax, paraffin, shea butter, cocoa seed butter, tocopherol, tocopherol acetate, spermaceti The invention relates to a novel composition comprising: a first embodiment of the present invention comprises one or more of stearyl alcohol, the first embodiment comprises one or more of carbomer, sodium carbomer, xanthan gum, polyacrylate polymers, and grafted starch, the second embodiment comprises one or more of emulsifiers, glyceryl stearate, PEG-100 stearate, polyglycerol emulsifiers, hydrogenated lecithin, and tri(laureth-4) phosphate, the second embodiment comprises one or more of preservatives, caprylylhydroxamic acid, p-hydroxyacetophenone, caprylyl glycol, ethylhexylglycerin, and phenoxyethanol, and the second embodiment comprises one or more of flavoring agents, including plant floral water, plant extracts, and plant essential oils.
10. The skin care product according to claim 8, characterized in that The skin care product is essence, cream or lotion.