CN112716855B

CN112716855B - Composition with barrier repairing and anti-aging effects and preparation method and application thereof

Info

Publication number: CN112716855B
Application number: CN202110114706.0A
Authority: CN
Inventors: 周秋娜; 金荣熙; 周春亚; 金炫延; 尹锺赫; 金延埈
Original assignee: Cosmax China Cosmetics Co Ltd
Current assignee: Cosmax China Cosmetics Co Ltd
Priority date: 2021-01-27
Filing date: 2021-01-27
Publication date: 2023-01-06
Anticipated expiration: 2041-01-27
Also published as: CN112716855A

Abstract

The invention provides a composition with barrier repair and anti-aging effects, and a preparation method and application thereof. The composition consists of the following components in percentage by mass: rosa multiflora flower water: rosa centifolia flower extract: hydrolyzed red algae extract = (10-90): 0.5-3): 1-6). The effect of the compositions mixed according to a certain proportion is better than that of a single component under the same concentration. The composition with barrier repair and anti-aging effects provided by the invention starts from 'internal maintenance and external repair', and prevents and improves the aging symptoms of skin from the dimensions of moisture preservation, oxidation resistance, brightening, tightening and the like. The composition provided by the invention not only retains the unique fragrance of rosa multiflora, but also realizes the reutilization of rosa multiflora residues, and accords with the green, natural and sustainable concept.

Description

Composition with barrier repairing and anti-aging effects and preparation method and application thereof

Technical Field

The invention belongs to the field of cosmetics, and particularly relates to a composition with barrier repair and anti-aging effects, and a preparation method and application thereof.

Background

Modern people are under multiple stresses, such as fast pace of life, irregular work and rest, environmental pollution and the like, and the young people of 20-25 years old are induced to be 'premature senility', and the symptoms of aging, such as fine lines, dry lines, darkness, dark circles, slight looseness of outlines and the like, begin to appear. Young people also have premature aging due to improper skin care.

At present, the anti-aging composition on the market mostly focuses on the skin repair after damage, and treats the symptoms rather than the root causes. As a layer of structure on the outermost side of the human body, the skin is the first barrier of the human body against external damage, and the importance is self-evident. The stratum corneum, the upper layer of the epidermis, is differentiated from keratinocytes to constitute different layers of epidermal cells, and thus the physiological function of the skin barrier depends on the process of cell maturation, i.e., the process of proliferative differentiation of keratinocytes, while Ca2+ plays a key role in epidermal differentiation. In the normal skin barrier, the distribution of Ca2+ on the surface layer of the skin is in a special T shape, and the closer to the surface layer of the skin, the higher the concentration of Ca2+ is. Research shows that in human beings, the content of Ca2+ (calcium ions) in the epidermis layer of the facial skin is the largest at the age of 13-19 years, the content of Ca2+ in the facial skin begins to decrease after the age of 20 years, when the concentration of Ca2+ is reduced, the maturation speed of keratinocytes is reduced, the stratum corneum cannot be continuously renewed, and the epidermis layer becomes thin, fragile, sensitive and loose, so that various problems occur to the skin.

From the biological point of view, aging is a time-oriented, naturally occurring life process, is the result of the combined action of many factors, is an irreversible life phenomenon, and simply lifting the skin barrier is far from sufficient. More and more researches show that the antioxidation is a key step for preventing skin aging, and because free radicals and oxidants can damage cell tissues and damage the normal metabolic action of the skin, the quantity of the free radicals is effectively controlled, and the method is particularly important for preventing skin aging. Therefore, there is a need to develop a composition that can repair the barrier and has antioxidant properties.

Flos rosae multiflorae (Rosa centifolia l.) is a shrub of Rosa genus of rosaceae family, also called pluronic rose, rosa multiflora, rosa grisea, rosa veneriformis, etc., is the first choice of perfume for luxury due to its unique fragrance, and also has excellent efficacy in skin care: has effects in keeping moisture, promoting melanin decomposition, resisting free radical, and delaying aging. The rosa multiflora flower residue contains rich nutrient components such as organic acid, polysaccharide, phenols, vitamins and the like, wherein the content of total flavonoids is up to 3.34 percent, and the rosa multiflora flower residue has a remarkable antioxidant effect. At present, the major purpose of rosa centifolia flowers is to serve as an extraction raw material of essential oil, a large amount of by-products, namely flower residues, can be generated, part of the flower residues are naturally air-dried to be used as fuel, and the rest of the flower residues are mostly used as garbage for treatment, so that resource waste and environmental pollution are caused. Therefore, how to utilize precious rosa multiflora efficiently and reasonably is very important.

Calcareous algae, also known as limnophyta, is a generic name for algae having the function of secreting or depositing calcium, most commonly known as Corallinaceae (Corallinaceae) of the Rhodophyta. Coral algae (coral algae) is a type of large calcified algae widely distributed in the ocean and is called as sarcodictyia glabra (organized/generic coral algae), sarcodictyia glabra (non-organized/non-generic coral algae), corallina shelllifera (CCA), corallina pellicularis (organizing coral algae), rhodochrous algae (rhodolith) and the like, depending on the difference in external morphology. The calcium algae are mostly formed in the sea and are rich in sea calcium, magnesium, trace elements and the like, wherein the calcium and magnesium elements play an important role in the protective barrier of the seaweed.

The existing product does not have the function of enhancing the epidermal barrier by regulating mineral elements in the epidermis, so that the composition which treats both symptoms and root causes and has the effects of barrier repair and anti-aging needs to be developed urgently, namely the composition which has the functions of regulating the mineral elements in the epidermis to enhance the epidermal barrier, repairing skin damage caused by external stimulation and has the effects of preventing and repairing.

Disclosure of Invention

In order to solve the defect that the anti-aging composition in the prior art can only be used for repairing aged skin but cannot adjust mineral elements in the epidermis to enhance the epidermal barrier function, the invention provides the composition with the effects of maintaining stability internally and repairing externally, namely, the composition can adjust the mineral elements in the epidermis to enhance the epidermal barrier function and repair skin damage caused by external stimulation and has the effects of preventing and repairing.

The application is realized by the following technical scheme:

the invention provides a composition with barrier repair and anti-aging effects, which comprises the following components in percentage by mass: the mass ratio of flos Rosae Davuricae water to flos Rosae Davuricae extract to hydrolyzed red algae extract is (10-90): (0.5-3): 1-6).

Preferably, the mass ratio of the rosa centifolia flower water to the rosa centifolia flower extract to the hydrolyzed red algae extract is (30-60): (0.5-1): 1-3).

Further preferably, the mass ratio of the rosa centifolia flower water to the rosa centifolia flower extract to the hydrolyzed red algae extract is 60.

The invention also provides a preparation method of the composition, which comprises the following steps:

1) Selecting fresh flos Rosae Davuricae, sorting, cleaning, drying at low temperature, and collecting condensate water to obtain flos Rosae Davuricae water;

2) Adding deionized water into the dried flowers in the step 1), performing wet superfine grinding, centrifuging, collecting supernatant, concentrating under reduced pressure, and drying the precipitate to obtain flos Rosae Davuricae extract;

3) Adding deionized water into red algae, heating to 90 deg.C, adding lactic acid, boiling, and maintaining for 2-4 hr; then filtering, and vacuum drying the filtrate to obtain hydrolyzed red algae extract;

4) Adding the obtained flos Rosae Davuricae extract and hydrolyzed red algae extract into flos Rosae Davuricae water, and mixing to obtain solution;

5) Filtering the solution obtained in the step 4) by using a microfiltration membrane and an ultrafiltration membrane in sequence, and collecting filtrate to obtain the composition.

Preferably, the low-temperature drying in step 1) is performed at a temperature lower than 50 ℃, and includes but is not limited to one or two of oven drying, freeze drying and vacuum drying.

Preferably, the dosage of the deionized water in the step 2) is 5-10 times of that of the dried flowers;

and 2) performing wet-process ultrafine grinding by using a colloid mill or a homogenizer.

Preferably, the dosage of the deionized water in the step 3) is 5-15 times of that of red algae;

the addition amount of lactic acid in the step 3) is 1-3 times of that of red algae;

in the step 5), the aperture of the microfiltration membrane is 0.1-1.0 μm, and the aperture of the ultrafiltration membrane is 0.01-0.05 μm.

Preferably, the method for blending in the step 5) is stirring or ultrasonic blending.

The invention also provides a cosmetic containing the composition; such cosmetics include, but are not limited to, lotions, creams, sprays, creams, masks, foundations.

Compared with the prior art, the application can obtain the following technical effects:

at the same total concentration, the DPPH clearance of the composition provided by the invention is higher than that of the single component, which indicates that 3 single raw materials can obtain better DPPH clearance effect in a combined mode. In addition, the invention provides a composition which uses rosa centifolia flower water to replace solvent water, reduces the use of the solvent water, uses flower residues to retain the unique fragrance of rosa centifolia, realizes the reutilization of the rosa centifolia flower residues, and accords with the green, natural and sustainable concept. The invention adopts microfiltration-ultrafiltration step-by-step filtration, realizes the purposes of separation, purification and refinement in a combined way, and has the advantages of simple and convenient operation, low energy consumption, short extraction time, better effect and more stable extract compared with the traditional method.

Drawings

FIG. 1 is a diagram showing an analysis of the expression of filaggrin FLG gene in Experimental example 3;

FIG. 2 is a diagram showing the analysis of the expression of the filaggrin degradation-associated gene CASP14 in Experimental example 3;

FIG. 3 is a graph showing moisture growth (%) of stratum corneum after using different compositions in Experimental example 5;

FIG. 4 is a graph showing the reduction (%) of the epidermal water loss value (%) after using different compositions in Experimental example 5;

FIG. 5 is a graph showing the reduction rate of melanin content after different compositions were used in Experimental example 5;

FIG. 6 is a comparison of the fine lines of the posterior canthus after the use of different compositions in Experimental example 5.

Detailed Description

Embodiments of the present application will be described in detail by examples, so that how to apply technical means to solve technical problems and achieve technical effects of the present application can be fully understood and implemented.

The raw materials and equipment used in the present application are all common raw materials and equipment in the field, and are all from commercially available products, unless otherwise specified. The methods used in this application are conventional in the art unless otherwise indicated.

There are many other possible embodiments of the present invention, which are not listed here, and the embodiments claimed in the claims of the present invention can be implemented.

In the following examples and comparative examples of the present invention, compositions having barrier repair, anti-aging effects were prepared using the following processes:

(1) Preparation of rosa centifolia flower water: selecting appropriate amount of fresh flos Rosae Davuricae, sorting, cleaning, drying at low temperature, condensing and collecting water vapor evaporated during drying to obtain flos Rosae Davuricae water;

(2) Preparing a rosa centifolia flower extract: taking a proper amount of flowers dried in the step (1), adding deionized water with the mass being 8 times that of the flowers, performing wet-process superfine grinding by using a colloid mill, centrifuging, taking supernate, performing reduced pressure concentration, and drying precipitates to obtain a rosa multiflora flower extract;

(3) Preparation of hydrolyzed red algae extract: taking a proper amount of red algae, adding deionized water with the mass of 5 times, heating to 90 ℃, adding lactic acid with the mass of 2 times, boiling, preserving heat for 3 hours, filtering, and drying filtrate in vacuum to obtain a hydrolyzed red algae extract;

(4) Adding the rosa centifolia flower extract and the hydrolyzed red algae extract obtained in the steps (2) and (3) into the rosa centifolia flower water obtained in the step (1) according to a certain proportion, and stirring and uniformly mixing;

(5) And (5) sequentially filtering the solution obtained in the step (4) by using a microfiltration membrane with the pore diameter of 0.45 mu m and an ultrafiltration membrane with the pore diameter of 0.05 mu m, and collecting filtrate to obtain the composition with the barrier repairing and anti-aging effects.

Table 1 shows the composition ratios of the compositions of examples 1 to 10 and comparative examples 1 to 3 (the composition ratios of multiflora rose water to multiflora rose flower extract to hydrolyzed red algae extract are by mass). Compositions of each example and comparative example were prepared according to table 1 and by the above-described method, and the resulting compositions were subjected to efficacy tests.

TABLE 1

Experimental example 1

Cytotoxicity evaluation test:

MTT assay is an experimental method widely used for detecting cytotoxicity or cell proliferation by measuring the number of live cells, and the detection principle is that succinate dehydrogenase in mitochondria of the live cells can reduce water-soluble yellow salt MTT (3- (4, 5-dimethylthiazole-2) -2, 5-diphenyl tetrazolium bromide) into water-insoluble blue formazan which is deposited in the cells, but dead cells do not have the function. The generated crystalline formazan is generally added with DMSO (dimethyl sulfoxide), and the absorbance is measured after dissolution. The specific experimental method is 1X10 in 96-well plate ⁴ Each 100. Mu.l of the culture medium was inoculated into a DMEM medium containing 10% bovine serum and keratinocytes (HaCaT) at a density of 100. Mu.l/well, and the medium was changed to a serum-free medium after 24 hours of culture. The culture was carried out for 24 hours after the treatment by adding the compositions at different concentrations to serum-free medium, respectively. Thereafter, the medium was removed, treated with 20. Mu.l of MTT solution, and allowed to react at 37 ℃ for 2 hours. 200 μ l of isopropanol was added to the cells from which the MTT solution was removed, gently shaken for 30min to completely dissolve the crystalline formazan, the absorbance was measured at 570nm, and the cell viability was calculated according to the following formula.

The blank was tested without the addition of composition. Table 2 shows the cytotoxicity results of the compositions obtained in examples 1 to 10 and comparative examples 1 to 3.

TABLE 2

Experimental example 2

And (3) antioxidant evaluation test:

DPPH is also called 1, 1-diphenyl-2-trinitrophenylhydrazine, is a very stable free radical with a nitrogen center, and the absolute ethanol solution of DPPH is purple, and when a free radical scavenger is added, the number of free radicals is reduced, the absorbance is reduced, the color of the solution is lightened, and the capability of scavenging the free radicals can be evaluated.

Measuring 40 mu l of each composition by using a pipettor, adding 160 mu l of DPPH ethanol solution (200 mu g/ml) for dilution and mixing uniformly, reacting for 30min at room temperature in a dark place after mixing uniformly, measuring absorbance A at 517nm, setting 3 groups of parallel experiments for each group to calculate the average value, wherein the calculation method is as follows:

clearance% = [1- (A) ₁ -A ₂ )/A ₀ ×100

In the formula: a. The ₀ Absorbance of DPPH solution without each composition added;

A ₁ adding each composition into DPPH solution and obtaining absorbance after reaction;

A ₂ the compositions were added to the DPPH solution and the absorbance before reaction was taken after mixing.

Table 3 shows the results of the antioxidant evaluation tests of the compositions of examples 1 to 10 and comparative examples 1 to 3.

TABLE 3

From the above table, it can be seen that:

1) The DPPH clearance of the compositions obtained in examples 1-10 is higher than that of comparative examples 1-3, which shows that 3 single raw materials can obtain better DPPH clearance effect after being combined in a certain proportion;

2) DPPH clearance and 3 raw materials all have certain quantitative effect relation, and after the mass fraction increases to a certain range, the rising trend slows down: comparative examples 1-4 show that the DPPH clearance rate of the compositions obtained in examples 3 and 4 is not large, which indicates that the effect is not obviously increased after the mass fraction of the rosa multiflora flower water is more than 60; examples 3, 5 and 6 compare and find that the DPPH clearance rate of the compositions with 3 and 6 parts by weight of the hydrolyzed red algae extract is not large; examples 7-10 comparison showed that the DPPH clearance differences were not significant for compositions with rosa centifolia flower extract fractions 1 and 3. Therefore, in combination with DPPH scavenging ability and economic efficiency, examples 5 (60.

In summary, the combination of the 3 raw materials in different proportions has different oxidation resistance, and is preferably rosa centifolia water: rosa centifolia flower extract: the mass percentage of the hydrolyzed red algae extract is (30-60): (0.5-1): 1-3), more preferably, the mass percentage of the rosa centifolia water: rosa centifolia flower extract: the mass percentages of the hydrolyzed red algae extract are 60.5.

Experimental example 3

Keratinocyte gene expression assay:

filaggrin (FLG) is an important condition for the terminal differentiation of keratinocytes, plays an important role in the differentiation and formation of epidermis, caspase14 (CASP 14, aspartase proteolytic enzyme) catalyzes the synthesis of FLG, and the combination of the FLG and the Caspase14 is an important condition for forming a skin surface barrier. Therefore, both of them can be used as an index for evaluating the state of the skin barrier function.

And detecting the regulation of the composition on the target gene expression level by adopting a real-time fluorescent quantitative PCR method.

6 groups of culture media with different compositions, which are identical to other components, are respectively arranged on a 6-well plate, wherein: negative control (no composition NT); positive control (containing 0.01% hyaluronic HA by mass); in two groups of culture media containing the composition obtained in example 5, the mass concentration of the composition is 0.1% and 0.3%, respectively; in both groups of the culture media containing the composition obtained in example 7, the mass concentrations of the compositions were 0.1% and 0.3%, respectively.

HaCaT cells were seeded at 3 × 105/wellCulturing on a 6-well plate for 24h, and growing the cells in an adherent way. Formulating the composition into gradient dilution spots of 1X, 2 concentrations, adding 2ml per well to the cell plate, 5% CO at 37% ₂ After the constant-temperature incubator is continuously incubated for 24 hours, cells are collected, total RNA of the cells is extracted by using a PureLink mini RNA extraction kit, the concentration of nucleic acid is quantitative, 200-800ng of RNA in each tube is used as a template, and the relative expression quantity (20 mu l system) of each gene is detected by performing Real-Time quantitative fluorescence PCR (reverse transcription + Real Time-PCR) reaction in a Taqman one-step method.

Calculating the formula:

Fold change＝2 ^-ΔΔCT ；

- Δ Δ CT = [ (CT target gene-CT reference gene) test group- (CT target gene-CT reference gene) control group ].

In this experiment, 2 target genes were FLG and CASP14, and 1 reference gene was GAPDH.

The results are shown in FIGS. 1-2, respectively.

Wherein:

1) FLG gene expression as shown in fig. 1, 0.3% of example 5 can significantly induce its expression, and the effect is stronger than that of hyaluronic acid, and is increased by about 1-fold compared to NT, and 0.3% of example 7 also significantly up-regulates its expression, but the effect is weaker than that of hyaluronic acid, and the order of up-regulated expression is 0.3% of example 5 > 0.01% of hyaluronic acid > 0.3% of example 7 > 0.1% of example 5 > 0.1% of example 7 > negative control;

2) Filaggrin degradation as shown in fig. 2, 0.3% example 5 significantly increased the expression of CASP14 gene and was more potent than hyaluronic acid, increasing it by about 4-fold compared to NT, and 0.3% example 7 also significantly upregulated its expression, but less potent than hyaluronic acid, with the order of upregulated expression being 0.3% example 5 > 0.01% hyaluronic acid > 0.3% example 7 > 0.1% example 5 > 0.1% example 7 > negative control.

Experimental example 4

And (3) testing safety patches:

dripping 20 μ l of the solution containing the composition to be tested into a plaque tester, wherein the control hole is blank control (pure water); the spot tester with the tested object is attached to the bent side of the forearm of the tested person, and the tested person is lightly pressed by the palm to be uniformly attached to the skin for 24 hours; the skin irritation and sensitization were observed as in table 4 after 30min, 24 hours, and 48 hours, respectively, after removal of the test article plaque test device, and the observation results were recorded. Skin-closed patch test skin response grading criteria are shown in table 4.

TABLE 4

The experimental results are as follows: the compositions obtained in comparative examples 5 and 7 were subjected to the human skin patch test and the results are shown in Table 5. The result of the skin patch test of the human body shows that 20 people have no adverse reaction to the skin.

TABLE 5

Experimental example 5

Screening 30 healthy volunteers, 15 males and 15 females, aged 22-35 years. An emulsion containing 1% by mass of the composition obtained in example 5, example 7 and comparative examples 1 to 3 was used to set a blank (using water) 2 times a day for 28 consecutive days. Meanwhile, in the test, the increase of stratum corneum moisture, decrease of the value of teal of transepidermal water loss, decrease of melanin content and fine lines of eyes were continuously recorded by using the instrument, and the results are respectively shown in fig. 3 to 6.

Wherein:

1) FIG. 3 is a graph of stratum corneum moisture growth (%) with different compositions; FIG. 4 is a graph showing the reduction (%) of the epidermal water loss value of Tewl values with different compositions. As can be seen from FIGS. 3 and 4, the compositions of examples and comparative examples, which improve skin moisture content and transdermal water loss, example 5 > example 7 > comparative example 3 > comparative example 1 > comparative example 2, demonstrate that hydrolyzed red algae extract has a positive effect on skin moisturization.

2) FIG. 5 is a graph of the rate of reduction of melanin content using different compositions. As can be seen from FIG. 5, the examples and comparative examples can reduce the melanin content of the skin, and the reduction trend is in the order of example 7 > example 5 > comparative example 2 > comparative example 1 > comparative example 2, which shows that the rosa centifolia flower extract has a positive effect on skin lightening.

3) Figure 6 is a comparison of canthus striae after use of different compositions. As can be seen from fig. 6, examples 5 and 7 have a significant effect of improving the fine lines of the eyes.

The details not described in the specification of the present application belong to the common general knowledge of those skilled in the art.

In the following description and in the claims, the terms "include" and "comprise" are used in an open-ended fashion, and thus should be interpreted to mean "include, but not limited to. "substantially" means within an acceptable error range, that a person skilled in the art can solve the technical problem within a certain error range to substantially achieve the technical effect.

It is also noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a good or system that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such good or system. Without further limitation, an element defined by the phrases "comprising one of \8230;" does not exclude the presence of additional like elements in an article or system comprising the element.

The foregoing description shows and describes several preferred embodiments of the present application, but as aforementioned, it is to be understood that the application is not limited to the forms disclosed herein, and is not to be construed as excluding other embodiments, but rather is capable of use in various other combinations, modifications, and environments and is capable of changes within the scope of the inventive concept as expressed herein, commensurate with the above teachings, or the skill or knowledge of the relevant art. And that modifications and variations may be effected by those skilled in the art without departing from the spirit and scope of the application, which is to be protected by the claims appended hereto.

Claims

1. The composition with the barrier repair and anti-aging effects is characterized by comprising the following components in percentage by mass: the mass ratio of the rosa centifolia flower water to the rosa centifolia flower extract to the hydrolyzed red alga extract is 60; the composition is prepared by the following steps:

1) Selecting fresh flos Rosae Davuricae, sorting, cleaning, drying at low temperature, and collecting water vapor evaporated during drying to obtain flos Rosae Davuricae water;

2) Adding deionized water into the flowers dried in the step 1), performing wet superfine grinding, centrifuging, collecting supernatant, concentrating under reduced pressure, and drying the precipitate to obtain flos Rosae Davuricae extract;

3) Adding deionized water into red algae, heating to 90 deg.C, adding lactic acid, boiling, and keeping the temperature for 2-4 hr; then filtering, and vacuum drying the filtrate to obtain hydrolyzed red algae extract;

2. The composition of claim 1,

the low-temperature drying condition in the step 1) is drying at a temperature lower than 50 ℃; the drying method is one or two of oven drying, freeze drying and vacuum drying.

3. The composition of claim 1,

the dosage of the deionized water in the step 2) is 5-10 times of that of the dried flowers;

4. The composition of claim 1,

the dosage of the deionized water in the step 3) is 5-15 times of that of the red algae;

the addition amount of the lactic acid in the step 3) is 1-3 times of that of the red algae;

in the step 5), the aperture of the micro-filtration membrane is 0.1-1.0 μm, and the aperture of the ultra-filtration membrane is 0.01-0.05 μm.

5. A cosmetic comprising the composition of claim 1; the cosmetic comprises water, emulsion, spray, cream, facial mask, and foundation solution.