A composition with a tightening effect, its preparation method and applications

Abstract

The present invention relates to a composition with a firming effect, its preparation method and application. The composition with a firming effect includes humanized collagen, copper peptide and red algae extract. Among them, the mass ratio of the humanized collagen, copper peptide and red algae extract is 0.5-5:0.1-2:0.5-3. The present invention uses human type III collagen with a 164.88° bend in the core functional region, which has a typical triple helix structure, has good skin affinity and is beneficial to improving the skin regeneration speed and restoring the skin barrier function. In addition, the present invention also uses red algae extract and copper peptide in combination with humanized collagen to promote collagen synthesis from multiple angles and firm the skin. Each raw material used in the present invention is mild and non-irritating, and is suitable for sensitive skin.

Description

Technical Field

[0001] The present invention belongs to the technical field of daily chemical products, and in particular relates to a composition with firming effect, its preparation method and application. Background Art

[0002] With the growth of age, most people will show signs of skin aging. Skin blood circulation will slow down, and the subcutaneous adipose layer will become loose and lack elasticity. Skin laxity and subcutaneous fat loss may also lead to contour changes such as sunken apple muscles and sunken temples, and the formation of eye bags, etc. Therefore, skin laxity is one of the major signals of aging, and how to firm the skin is a difficult problem to solve.

[0003] Chinese Patent CN115252508A discloses a firming essence and its preparation method, including 92-97 parts of Phase A, 0.5-1 part of Phase B and 2-7.5 parts of Phase C; the Phase A includes: 80-88 parts of solvent, 8-12 parts of humectant A, 0.05-0.15 parts of thickener, and 0.5-1.8 parts of skin conditioner A. The Phase B includes: 0.2-0.8 parts of emollient and 0.1-0.4 parts of emulsifier; the Phase C includes 2-3 parts of humectant B, 0.1-0.4 parts of skin conditioner B, 0.1-0.4 parts of skin conditioner C, and 0.1-0.4 parts of skin conditioner D. In this patent, a conditioner is composed of various extracts (such as Tremella fuciformis extract, Paeonia suffruticosa root extract, Eriobotrya japonica leaf extract, Selaginella tamariscina extract, Platycladus orientalis leaf extract, Paeonia lactiflora extract, Tilia europaea flower extract, Saussurea tinctoria extract, Althaea officinalis root extract, etc.) and oligopeptides. The essence obtained through reasonable cooperation can penetrate into the basal layer of the skin, absorb and retain skin moisture, form a protective film on the skin surface, slow down the loss of skin moisture, and at the same time has the effect of tightening the skin.

[0004] Chinese Patent CN111150701A discloses an anti-wrinkle firming essence and its preparation method. In this patent, the anti-wrinkle firming composition, by weight, includes: 0.53 parts of Spirulina maxima extract, 0.005-0.1 parts of sodium hyaluronate, 0.01-0.05 parts of cyclodextrin, 0.01-0.05 parts of ubiquinone, 0.1-1 part of avenanthramide, 1-5 parts of yeast extract, 0.1-1 part of fucoidan, 0.01-0.1 parts of humic acid, 0.1-3 parts of Hylocereus undatus fruit extract, 0.05-0.3 parts of decapeptide-4, 0.05-0.3 parts of glycoprotein, and 0.005-0.05 parts of rose flower oil. It can quickly penetrate deep into the skin to replenish moisture, smooth the skin, smooth out fine lines and wrinkles, endow the skin with elastic firmness, delay skin aging, promote the regeneration of young cells, and make the skin present brightness and luster.

[0005] However, in the solutions disclosed in the above patents, the small molecule polypeptides directly extracted from natural organisms or obtained by protease hydrolysis have disadvantages such as poor stability, short half-life, and difficulty in improving purity.

[0006] In addition, most skin care products on the market can meet basic requirements such as whitening and moisturizing, but they cannot deeply improve the skin, promote skin metabolism, repair the skin, and achieve the effect of firming the skin. Some existing firming skin care products will add hydrolyzed collagen, hyaluronic acid, polypeptides or retinol, etc. However, the molecular weight of hydrolyzed collagen is too large and it is not easy to penetrate the skin barrier of the human body to reach the dermis layer. Hyaluronic acid cannot essentially reduce the loss of collagen. Polypeptides and retinol have certain irritation and safety risks to the skin. At the same time, some firming skin care products overly emphasize the efficacy of the products, and the added ingredients are highly irritating, resulting in a poor use experience. Summary of the Invention

[0007] In order to improve the problems of poor effect of promoting collagen synthesis, irritation or safety risks existing in the existing firming skin care products, the present invention provides a composition with firming efficacy, a preparation method thereof and applications.

[0008] The object of the present invention can be achieved by the following technical solutions:

[0009] In a first aspect, the present invention provides a composition with firming efficacy, comprising humanized collagen, copper peptide and red algae extract, wherein the mass ratio of the humanized collagen, copper peptide and red algae extract is 0.5-5:0.1-2:0.5-3.

[0010] In an embodiment of the present invention, preferably, the mass ratio of the humanized collagen, copper peptide and red algae extract is 2-5:1-2:1.5-3; more preferably, the mass ratio of the humanized collagen, copper peptide and red algae extract is 3:1:1.5.

[0011] In an embodiment of the present invention, the humanized collagen is human type III collagen, the central functional region of which is bent at 164.88° and has a clear triple helix structure. Its cell adhesion is 183% of that of human own collagen, and it can reconstruct the three-dimensional structure fiber network of type III collagen, activate the extracellular matrix, support the orderly growth of various cells, and give full play to the functions of cell repair, regeneration and anti-aging.

[0012] In an embodiment of the present invention, the chemical structure of the copper peptide is as follows:

[0013] .

[0014] The copper peptide is Pal-GHK-Cu with the amino acid sequence of H-Gly-His-Lys (Pal)-OH.(Cu), and its chemical formula is C 30 H 52 CuN5O5.

[0015] In an embodiment of the present invention, the preparation method of the copper peptide includes the following steps: mixing Pal-GHK and copper acetate in water and reacting to synthesize a copper peptide with the amino acid sequence of H-Gly-His-Lys (Pal)-OH.(Cu).

[0016] In an embodiment of the present invention, the preparation method of the copper peptide includes the following steps:

[0017] I. Preparation of Pal-GHK

[0018] (1) Amino protection

[0019] Under stirring, add Fmoc-Lys (Dde)-OH and an amino acid containing 9-fluorenylmethoxycarbonyl (Fmoc) or tert-butylcarbonyl (Boc) into the reactor, and stir and react at room temperature for 1-3 h for amino protection to obtain GHK(Pal);

[0020] (2) Palmitic acid modification

[0021] Under stirring, continue to add palmitic acid into the reactor, and stir and react at room temperature for 1-3 h for side chain modification;

[0022] The amino acid containing 9-fluorenylmethoxycarbonyl (Fmoc) or tert-butylcarbonyl (Boc) is Fmoc-His(Trt)-OH or Boc-Gly-OH;

[0023] When the amino acid is Fmoc-His(Trt)-OH, the mass ratio of the amounts used of Fmoc-Lys(Dde)-OH, Fmoc-His(Trt)-OH, and palmitic acid is: 2-4: 2-4: 0.5-2;

[0024] When the amino acid is Boc-Gly-OH, the mass ratio of the amounts used of Fmoc-Lys (Dde)-OH, Boc-Gly-OH, and palmitic acid is: 2-4: 0.5-1: 0.5-2.

[0025] (3) Deprotection

[0026] The product obtained in step (2) is successively eluted with eluents (20% piperidine or N,N-dimethylformamide, and 2% hydrazine or N,N-dimethylformamide) (i.e., the product is first mixed with 20% piperidine or N,N-dimethylformamide and stirred for 1 - 3 h, then centrifuged to remove the Fmoc group, and then mixed with 2% hydrazine or N,N-dimethylformamide and stirred for 1 - 3 h, then centrifuged to remove the Dde group), to remove the Fmoc group and the Dde group;

[0027] (4) Amino acid residue elongation

[0028] The product obtained in step (3) is added with an active reagent and reacted at room temperature for 1 - 3 h to elongate the carbon chain of the amino acid residue; the active reagent includes N-hydroxybenzotriazole and N,N'-diisopropylcarbodiimide, and the dosage of the active reagent is 1 - 3 times the molar amount of Fmoc-Lys (Dde)-OH.

[0029] (5) Separation and purification

[0030] The product obtained in step (4) is successively washed with dimethylformamide, dichloromethane and methanol, dried in vacuo, and then reacted with an aqueous solution containing trifluoroacetic acid and triisopropylsilane (a mixture of trifluoroacetic acid, triisopropylsilane and water with a volume ratio of 90:5:5) to separate the product from the resin. The product is concentrated, centrifuged, and then purified and separated by reverse-phase high performance liquid chromatography to obtain the Pal-GHK product.

[0031] II. Synthesis of Pal-GHK-Cu

[0032] Pal-GHK is mixed and reacted with copper acetate in water, and the pH of the reaction solution is adjusted to 4 - 6 with sodium hydroxide. The precipitated sky-blue solid product is washed with water and dried in vacuo to obtain a copper complex with the amino acid sequence H-Gly-His-Lys (Pal)-OH.(Cu).

[0033] In the present invention, the copper peptide is an active peptide with skin repair effect, which can be used as a remodeling agent for cell activation to help the skin restore its self-repair ability; promote the production of collagen and reduce the growth of wrinkles; it is also applicable to anti-inflammatory repair, promote hair growth, reduce hair loss, and has broad application prospects.

[0034] In one embodiment of the present invention, the red algal extract is an extract from Botryocladia leptopoda from Taiwan. Botryocladia leptopoda is mainly distributed in the waters through which the warm Kuroshio current passes, and its vesicles contain a variety of marine essence active substances. Therefore, the red algal extract in the present invention contains a variety of marine essence active substances, such as polysaccharides, polypeptides, phenols, terpenoids, etc., which has the effect of promoting water circulation, promoting the movement of abundant moisture in the skin base layer to the skin surface, achieving soft, moist and transparent skin, and contributing to skin health.

[0035] In one embodiment of the present invention, humanized collagen, copper peptide and red algal extract are placed in a mixed solution of butylene glycol and methyl palmitate. The mass ratio of humanized collagen, copper peptide, red algal extract, butylene glycol and methyl palmitate is 0.5 - 5:0.1 - 2:0.5 - 3:5 - 10:0.1 - 0.2. At this time, the composition with firming effect is in the form of a microemulsion. The droplets of the microemulsion are only nanosized. Among them, methyl palmitate is dissolved in butylene glycol to form a pre - preparation, and this pre - preparation is mixed with humanized collagen, red algal extract and copper peptide, and the active ingredients humanized collagen, copper peptide and red algal extract are wrapped in the pre - preparation. By using this microemulsion encapsulation technology, while increasing the solubility of the active ingredients, it can also improve their permeability. In the present invention, the composition with firming effect adopts the form of a microemulsion, and these active ingredients can be dispersed in the microemulsion; the surfactant of the microemulsion can reduce the diffusion barrier of the stratum corneum; the droplets of the microemulsion are only nanosized and can easily transport the loaded active ingredients through the stratum corneum of the skin.

[0036] Second, the present invention provides a preparation method of a composition with firming effect, which mixes humanized collagen, red algal extract and copper peptide evenly to obtain a composition with firming effect.

[0037] Furthermore, the present invention provides a preparation method of a composition with firming effect, including the following steps:

[0038] Weigh methyl palmitate and dissolve it in butylene glycol at 80 - 85 °C. After complete dissolution, cool it to room temperature, and then add humanized collagen, red algal extract and copper peptide to it and mix evenly to obtain a composition with firming effect. Preferably, the temperature for adding humanized collagen, red algal extract and copper peptide for mixing is 30 - 45 °C, and the mixing time is 10 - 20 min.

[0039] Third, the present invention provides an application of a composition with firming effect, using the composition with firming effect in skin care products or cosmetics. The skin care products or cosmetics include essence, lotion, cream, etc.

[0040] Fourthly, the present invention provides a skin care product or cosmetic containing a composition with firming effect, which contains a composition with firming effect. Among them, the mass percentage content of humanized collagen in the skin care product or cosmetic is 0.5 - 5%, the mass percentage content of red algae extract in the skin care product or cosmetic is 0.5 - 3%, and the mass percentage content of copper peptide in the skin care product or cosmetic is 0.1 - 2%.

[0041] Furthermore, the present invention also provides a serum with firming effect. The components and contents of the serum with firming effect are as follows:

[0042]

[0043] Furthermore, the present invention also provides a preparation method of the serum with firming effect, which includes the following steps:

[0044] (1) Weigh each substance in phase A and mix them evenly at room temperature to obtain mixture A;

[0045] (2) Weigh methyl palmitate and dissolve it in butanediol at 80 - 85°C. After complete dissolution, cool it to room temperature, and then add humanized collagen, red algae extract and copper peptide to it, and mix them evenly to obtain mixture B;

[0046] (3) Under stirring conditions, add mixture B to mixture A. After stirring evenly, add each substance in phase C and stir evenly to obtain the serum with firming effect.

[0047] Compared with the prior art, the beneficial effects of the present invention include:

[0048] (1) The present invention uses humanized type III collagen with a 164.88° bend in the core functional region, which has a typical triple helix structure, has good skin affinity and is beneficial to improving the skin regeneration speed and restoring the skin barrier function. In addition, the present invention also uses red algae extract and copper peptide in combination with humanized collagen to promote collagen synthesis from multiple angles and firm the skin.

[0049] (2) In a preferred embodiment of the present invention, humanized collagen, copper peptide and red algal extract are placed in a mixed solution of butanediol and methyl palmitate. The mass ratio of humanized collagen, copper peptide, red algal extract, butanediol and methyl palmitate is 0.5-5:0.1-2:0.5-3:5-10:0.1-0.2. At this time, the composition with firming effect is in the form of a microemulsion, and the droplets of the microemulsion are only nanosized. Among them, methyl palmitate is a surfactant that can reduce the interfacial tension between oil and water; butanediol is a co-surfactant that can change the spontaneous curvature and elasticity of the surfactant interfacial film, thereby improving the stability of the microemulsion. Methyl palmitate dissolves in butanediol to form a special preparation, and this preparation is mixed with humanized collagen, red algal extract and copper peptide, and the active ingredients humanized collagen, copper peptide and red algal extract are wrapped in the surfactant. By using this microemulsion encapsulation technology, while increasing the solubility of the active ingredients, it can also improve their permeability. In the present invention, the composition with firming effect is in the form of a microemulsion, and these active ingredients can be dispersed in the microemulsion; the surfactant of the microemulsion can reduce the diffusion barrier of the stratum corneum; the droplets of the microemulsion are only nanosized and can easily transport the loaded active ingredients through the stratum corneum of the skin.

[0050] (3) Each raw material used in the present invention is mild and non-irritating, and is suitable for sensitive skin. Description of the Drawings

[0051] Figure 1 It is the HPLC chart of Pal-GHK-Cu;

[0052] Figure 2 It is the mass spectrometry chart of Pal-GHK-Cu;

[0053] Figure 3 It is the 3D rendering of Pal-GHK-Cu;

[0054] Figure 4 It is the skin firming F4 value of the subjects using the essence sample of Example 1;

[0055] Figure 5 It is the change value of the skin firming F4 value of the subjects using the essence sample of Example 1;

[0056] Figure 6 It is the change rate of the skin firming F4 value of the subjects using the essence sample of Example 1;

[0057] Figure 7 It is the comparison result of the mouse skin retention amount of the essence samples of Example 1 and Comparative Examples 1-8. Detailed Description of the Invention

[0058] The technical solutions of the present invention will be further described below through specific embodiments. Those skilled in the art should understand that the embodiments are only for helping to understand the present invention and should not be regarded as specific limitations to the present invention. Some non-essential modifications and adjustments made according to the present invention still fall within the protection scope of the present invention. Unless otherwise specified, the following percentages (%) are all mass percentages.

[0059] In the following examples, the chemical structure of the copper peptide is as follows:

[0060] 。

[0061] The copper peptide is Pal-GHK-Cu with the amino acid sequence of H-Gly-His-Lys (Pal)-OH.(Cu), and its chemical formula is C 30 H 52 CuN5O5.

[0062] The preparation method of the copper peptide includes the following steps:

[0063] I. Preparation of Pal-GHK

[0064] (1) Amino protection

[0065] Under stirring, add Fmoc-Lys (Dde)-OH resin and Boc-Gly-OH into the reactor and stir at room temperature for 2.0 h for amino protection to obtain GHK(Pal);

[0066] (2) Palmitic acid modification

[0067] Under stirring, continue to add palmitic acid into the reactor and stir at room temperature for 2.0 h for side chain modification;

[0068] The dosage ratio of Fmoc-Lys (Dde)-OH, Boc-Gly-OH and palmitic acid is: mass ratio 3.37:1:1.46.

[0069] (3) Deprotection

[0070] Elute the product obtained in step (2) successively with an eluent (20% piperidine and 2% N,N-dimethylformamide) (that is, first mix the product with 20% piperidine and stir for 2.0 h and then centrifuge to remove the Fmoc group; then mix with 2.0% N,N-dimethylformamide and stir for 2.0 h and then centrifuge to remove the Dde group) to remove the Fmoc group and the Dde group;

[0071] (4) Amino acid residue extension

[0072] The product obtained in step (3) was added with the active reagents N-hydroxybenzotriazole and N,N′-diisopropylcarbodiimide), and reacted at room temperature for 2.0 h to elongate the carbon chain of the amino acid residue; the dosage of the active reagents was 2.0 times the molar amount of Fmoc-Lys (Dde)-OH resin.

[0073] (5)Separation and purification

[0074] The product obtained in step (4) was washed successively with dimethylformamide 6 times, dichloromethane 6 times, and methanol 2 times, and then dried under vacuum. Then, the resin attached with the product was reacted with an aqueous solution containing trifluoroacetic acid and triisopropylsilane (a mixture of trifluoroacetic acid, triisopropylsilane, and water with a volume ratio of 90:5:5) to separate the product from the resin. The product was concentrated to a residue, precipitated with cold diethyl ether and centrifuged, and then the solid was washed with cold diethyl ether 3 times and centrifuged, and dried under vacuum. Then, reverse-phase high-performance liquid chromatography (column: Unisil, C18, 10 μ, 120 Å, 250 x 50 mm) was used to purify and separate the peptide to obtain the Pal-GHK product.

[0075] II. Synthesis of Pal-GHK-Cu

[0076] Pal-GHK was mixed and reacted with copper acetate in water, and the pH of the reaction solution was adjusted to 4 - 6 with sodium hydroxide. The precipitated sky-blue solid product was washed 3 times with water and dried under vacuum to obtain a copper complex with the amino acid sequence H-Gly-His-Lys (Pal)-OH.(Cu).

[0077] (6)The prepared Pal-GHK-Cu was characterized by high-performance liquid chromatography (Agilent, 1100) and mass spectrometry (Agilent, 6120). As Figure 1-2 shown, it can be seen from the figure that under these analysis conditions, the prepared Pal-GHK-Cu has good resolution, high ultraviolet absorbance, a relatively symmetric peak shape, a retention time of about 12 minutes, and a molecular weight of 640.33 g / mol.

[0078] The 3D rendering of Pal-GHK-Cu is as shown in Figure 3 shown. Safety tests were carried out on the copper peptide

[0079] According to Sections 6.4, 6.5, and 6.6 of the "Technical Specifications for Cosmetics Safety" (2015 Edition), the skin application safety of the copper peptide was evaluated. As shown in Tables 1 and 2:

[0080] Table 1 Results of cell proliferation assay (human keratinocytes)

[0081]

[0082] Table 2 Results of Cell Proliferation Assay (Human Fibroblasts)

[0083]

[0084] In the above table, SC indicates: negative control;

[0085] SD indicates: relative migration rate;

[0086] PC indicates: positive control;

[0087] It can be seen that after treating keratinocytes with 25 μg / L of copper peptide for 24 hours, the proliferation increased by nearly 200%, while the proliferation of copper peptide cells increased by nearly 20%. The analysis results further showed that copper peptide has better epidermal repair ability. As can be seen from Table 3, after treating with 25 μg / L of copper peptide for 24 hours, the proliferation rate of fibroblasts increased by more than 300%, which is better than the proliferation rate after treating with copper peptide for 24 hours. This copper peptide can maintain the proliferation activity of fibroblasts and has a potential effect on dermal repair.

[0088] Table 3 Results of Cell Migration

[0089] Sample Name Average Mobility Average Relative Mobility Relative Mobility SD P Value SC 0.25 1.00 0.126 / PC(10% FBS) 0.39 1.53 0.178 0.000 ** 25 μg / l Pal-GHK-Cu 0.31 1.22 0.102 0.008 ** 25 μg / l GHK-Cu 0.27 1.06 0.094 0.352

[0090] It can be seen that after treating with 25 μg / L of copper peptide for 24 hours, the relative migration rate of fibroblasts increased significantly (p < 0.01). Compared with copper peptide, copper peptide has better cell migration ability, further indicating that copper peptide has better skin repair ability.

[0091] In the following examples, the red algal extract is an extract from Botryococcus braunii from Taiwan and can be directly purchased.

[0092] In the following examples, the humanized collagen is human type III collagen and can be directly purchased.

[0093] In the following examples, other raw materials are all common raw materials for daily chemicals and can be directly purchased.

[0094] Examples 1, 2, 3 and Comparative Examples 1 - 8

[0095] Examples 1, 2, 3 and Comparative Examples 1 - 8 are essence liquids composed of different components, and the raw material component ratios of Example 1 and Comparative Examples 1 - 8 are shown in Table 4.

[0096] Table 4 Raw Material Component Ratios of Example 1 and Comparative Examples 1 - 8 (Weight Percentage)

[0097]

[0098] The preparation process flow of the essence in Examples 1, 2, and 3 is as follows:

[0099] (1) Weigh each substance in Phase A according to the weights in Table 4 and mix them evenly at room temperature to obtain Mixture A.

[0100] (2) Weigh methyl palmitate and dissolve it in butanediol at 80 - 85 °C. After complete dissolution, cool it to room temperature, and then add humanized collagen, red algal extract, and copper peptide to it, and mix evenly to obtain Mixture B.

[0101] (3) Under stirring conditions, add Mixture B to Mixture A. After stirring evenly, add each substance in Phase C and stir evenly to obtain the essence in Examples 1, 2, and 3.

[0102] The preparation process flow of the essence in Comparative Examples 1 - 8 is as follows:

[0103] (1) Weigh each substance in Phase A according to the weights in Table 4 and mix them evenly at room temperature to obtain Mixture A.

[0104] (2) Weigh methyl palmitate and dissolve it in butanediol at 80 - 85 °C. After complete dissolution, cool it to room temperature, and then add humanized collagen, red algal extract or copper peptide to it, and mix evenly to obtain Mixture B.

[0105] (3) Under stirring conditions, add Mixture B to Mixture A. After stirring evenly, add each substance in Phase C and stir evenly to obtain the essence in Comparative Examples 1 - 8.

[0106] Test on the firming effect of the essence with different component compositions in Example 1 and Comparative Examples 1 - 8

[0107] (1) The skin elasticity measuring instrument Cutometer dual MPA580 is used to detect skin firmness. The method is as follows:

[0108] The research subjects were 330 Chinese healthy women with sensitive skin, aged between 25 and 50 years old, with an average age of 35.06 years. They were divided into 11 groups, with 30 subjects in each group. The subjects gave written consent under the condition of being informed. The subjects were willing to cooperate and understand the necessity and duration of the control in order to comply with the protocol formulated by the clinical trial center. For each group of subjects, the essence liquids of Examples 1-3 and Comparative Examples 1-8 were used on one half of the face respectively, and only water was used on the other half of the face. It was used once in the morning and once in the evening every day for 28 consecutive days. At D0, D14, and D28, the experimenter used the skin elasticity measuring instrument Cutometer dual MPA580 to detect skin firmness. A smaller F4 measurement value indicates an improvement in skin firmness. The data was statistically analyzed using SPSS 28.0. The test data was subjected to a normal distribution test. If the test data was normally distributed, the T-test method was used for statistical analysis; if the test data was non-normally distributed, the rank sum test method was used for analysis. Rank sum test was used for statistical analysis of rank data. The significance level of the statistical method was p < 0.05.

[0109] Among them, the average skin firmness F4 values of 30 subjects using the essence liquid sample of Example 1 are as Figure 4 shown, and the change values of the average skin firmness F4 values of the subjects using the essence liquid sample of Example 1 are as Figure 5 shown. The change rate of the average skin firmness F4 values of the subjects using the essence liquid sample of Example 1 is as Figure 6 shown. Compared with before using the essence liquid sample of Example 1, after using the essence liquid sample of Example 1 for 14 days, the skin firmness F4 decreased by 12.94%; after using the sample for 28 days, the skin firmness F4 value decreased by 26.22%, proving that continuous use of this essence liquid can effectively improve skin firmness.

[0110] The average skin firmness F4 changes of 30 subjects after using the essence liquid samples of Examples 2 and 3 are shown in Table 5.

[0111] Table 5 Average skin firmness F4 changes of 30 subjects after using the essence liquid samples of Examples 1, 2, and 3

[0112]

[0113] (2) In vitro percutaneous penetration experiment

[0114] Take healthy SPF-grade male mice, with a batch purchase quantity of 33. Decapitate and sacrifice healthy mice weighing 20 - 25 g, peel off the skin, depilate with sodium sulfide, remove subcutaneous fat, wash with water, then soak in normal saline for 30 min for standby. Randomly select mouse skins and ensure that the skin thickness is similar. After selection, fix them between the diffusion cell and the receiving cell, with the dermis layer of the skin facing the receiving cell and the stratum corneum facing the diffusion cell. The volume of the receiving cell is 15 ml, and the dermis layer of the skin should just contact the solvent in the receiving cell to prevent the generation of air bubbles. The receiving solution is freshly prepared normal saline, and the test solution uses distilled water as the solvent, adding 1 g of the sample (Examples 1 - 3 or Comparative Examples 1 - 8). Place the receiving cell on the magnetic stirrer of the intelligent transdermal experiment instrument, set the water bath temperature at 37°C, the stirring speed at 350 r / min, and the transdermal time at 12 h. After the experiment, remove the skin, wipe and rinse the surface of the mouse skin with normal saline. After drying at room temperature on filter paper, transfer the mouse skin to a small beaker, cut the mouse skin into pieces with scissors, add distilled water, ultrasonically extract for 30 min, and soak for 24 hours. After the extract is filtered through a 0.22 μm microporous membrane, chromatographic analysis is carried out to calculate the skin retention rate of different samples.

[0115] When conducting the above experiment, 33 mice were divided into 11 groups with 3 mice in each group, and the samples of Examples 1 - 3 and Comparative Examples 1 - 8 were added to each group respectively.

[0116] Skin retention amount Q (μg / cm 2 ), and the calculation formula is Q = Vc / A, where A is the effective diffusion area, V is the total volume of the skin extract, and c is the mass concentration of the sample in the skin extract (referring to the mass concentration of the essence of Examples 1 - 3 or Comparative Examples 1 - 8).

[0117] Figure 7 For the comparison of the average value of the skin retention amount of Examples 1 - 3 and Comparative Examples 1 - 8, the higher the skin retention amount, the better the permeability. Therefore, the corresponding essence has a better absorption effect and a better firming effect. From the experimental results, it can be seen that the permeability of Example 1 is significantly stronger than that of Comparative Examples 1 - 8. Among them, in Comparative Example 8, since it does not contain methyl palmitate and butanediol, it does not form a microemulsion state, and the skin retention amount of its solution is relatively low.

[0118] (3) Safety test

[0119] The human patch test was carried out with reference to the "Technical Specifications for Cosmetics Safety", GB17149.1-1997 "Diagnostic Criteria and Management Principles for Cosmetic Dermatoses", and GB17149.2-1997 "Diagnostic Criteria and Management Principles for Cosmetic Contact Dermatitis". 30 volunteers were recruited in this study. The inclusion criteria for the volunteers were as follows: 1) Aged between 15 and 60 years old, with a male-to-female ratio of 1:1; 2) Without serious diseases, immune deficiency or autoimmune diseases, and the test sites have not been treated for skin diseases or for beauty purposes; 3) Without active allergic diseases; 4) No anti-inflammatory drugs have been applied to the test sites in the past two months; 5) Those who have used antihistamines in the past week or have not used immunosuppressants in the past month; 6) Those who have not undergone patch tests in the past month. The informed consent form for the volunteers was signed. First, serial numbers were marked on the backs of all patch testers, and an equal amount of Example 1 was added. The processed patch testers were applied to the flexor side of the forearms of the subjects, and the skin conditions were observed at 24 and 48 hours respectively. The grading of the skin reaction after patch testing is shown in Table 6.

[0120] Table 6 Grading of Skin Reaction after Patch Testing

[0121]

[0122] The results showed that the scores of the samples of Example 1 were both 0 at 24 hours and 48 hours, and no skin adverse reactions occurred, indicating that the composition and essence product involved in the present invention are safe and non-irritating to the skin.

[0123] The above description of the embodiments is for the convenience of those of ordinary skill in the art to understand and use the invention. It is obvious that those who are familiar with the technology in this field can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative labor. Therefore, the present invention is not limited to the above embodiments, and the improvements and modifications made by those skilled in the art without departing from the scope of the present invention according to the disclosure of the present invention should be within the protection scope of the present invention.

Claims

1. A composition with a firming effect, characterized in that, It includes humanized collagen, copper peptide and red algae extract. The humanized collagen, copper peptide and red algae extract are placed in a mixed solution of butylene glycol and methyl palmitate. The mass ratio of humanized collagen, copper peptide, red algae extract, butylene glycol and methyl palmitate is 0.5 - 5:0.1 - 2:0.5 - 3:5 - 10:0.1 - 0.2; The humanized collagen is human type III collagen, whose central functional region is bent at 164.88° and has a triple helix structure; The chemical structure of the copper peptide is as follows: 。 2. The composition with a firming effect according to claim 1, characterized in that, The mass ratio of the humanized collagen, copper peptide and red algae extract is 2 - 5:1 - 2:1.5 - 3.

3. The composition with a firming effect according to claim 1, characterized in that, The mass ratio of the humanized collagen, copper peptide and red algae extract is 3:1:1.

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4. A method for preparing the composition with a firming effect according to claim 1 or 2 or 3, characterized in that, It includes the following steps: Weigh methyl palmitate and dissolve it in butylene glycol at 80 - 85°C. After complete dissolution, cool it to room temperature. Then add humanized collagen, red algae extract and copper peptide to it and mix evenly to obtain a composition with firming effect.

5. Use of the composition with a firming effect according to any one of claims 1-3, characterized in that, The composition with firming effect is used for preparing cosmetics.

6. A cosmetic containing the composition with a firming effect as described in any one of claims 1-3, characterized in that, It contains a composition with firming effect. Among them, the mass percentage content of humanized collagen in the cosmetics is 0.5 - 5%, the mass percentage content of red algae extract in the cosmetics is 0.5 - 3%, and the mass percentage content of copper peptide in the cosmetics is 0.1 - 2%.

7. An essence with a firming effect, characterized in that, The components and mass percentage contents of the essence with firming effect are as follows: Jojoba oil 0.5 - 3% Glycerol 1 - 10% Sweet almond oil 0.5 - 3% Glycerol monooleate 2 - 4% Butylene glycol 5 - 10% Methyl palmitate 0.1 - 0.2% Humanized collagen 0.5 - 5% Red algae extract 0.5 - 3% Copper peptide 0.1 - 2% Sodium hyaluronate 0.1 - 1% Xanthan gum 0.5 - 3% Triethanolamine 0.1 - 1% Fragrance 0.01 - 0.3% Deionized water the balance; The humanized collagen is human type III collagen, whose central functional region is bent at 164.88° and has a triple helix structure; The chemical structure of the copper peptide is as follows: 。 8. The preparation method of the essence with a firming effect according to claim 7, characterized in that, It includes the following steps: (1) Weigh jojoba oil, glycerol, sweet almond oil and glycerol monooleate by weight parts and mix them evenly at room temperature to obtain mixture A; (2) Weigh methyl palmitate and dissolve it in butylene glycol at 80 - 85°C. After complete dissolution, cool it to room temperature. Then add humanized collagen, red algae extract and copper peptide to it and mix evenly to obtain mixture B; (3) Under stirring conditions, add mixture B to mixture A. After stirring evenly, add sodium hyaluronate, xanthan gum, triethanolamine, fragrance and deionized water and stir evenly to obtain the essence with firming effect.

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