Anti-inflammatory and anti-aging composition, preparation method thereof and application of anti-inflammatory and anti-aging composition in skin care products

Abstract

The invention relates to an anti-inflammatory and anti-aging composition, a preparation method of the anti-inflammatory and anti-aging composition and application of the anti-inflammatory and anti-aging composition in skin care products, and belongs to the technical field of skin care products. The sea cucumber oligopeptide and the cysteine are cross-linked through the beta-cyclodextrin to form the anti-inflammatory and anti-aging composition. The active ingredients of the sea cucumber oligopeptide and cysteine in the composition can achieve synergistic interaction through a cross-linking reaction, and the composition is good in stability and especially can inhibit skin inflammatory aging in a targeted mode.

Description

Technical Field

[0001] The invention belongs to the technical field of skin care products, and in particular relates to an anti-inflammatory aging composition, a preparation method thereof, and application thereof in skin care products. Background Art

[0002] In modern skin care technology, the research and development of low molecular weight complexes has become an important means to enhance the efficacy of active ingredients and improve skin care effects. With the in-depth study of the mechanism of skin aging, the demand for anti-aging products increasingly emphasizes improving the stability of active ingredients and the sustained release effect.

[0003] As a natural polypeptide extracted from sea cucumbers, sea cucumber oligopeptides have significant anti-aging, anti-oxidation, and skin repair effects. Sea cucumber oligopeptides can promote collagen synthesis, enhance skin elasticity, reduce fine lines and wrinkles, and show strong effects in repairing skin barriers and resisting UV damage. However, sea cucumber oligopeptides are easily oxidized when in contact with air, resulting in unstable efficacy and affecting their long-term effects in skin care, which limits their effective application in skin care products.

[0004] Cysteine ​​is a sulfur-containing amino acid that is widely used in skin moisturizing, anti-oxidation, and promoting skin cell repair. Cysteine ​​can neutralize free radicals in the skin and reduce oxidative stress. Its mechanism of action in the field of skin care is mainly through enhancing the skin barrier function and improving the skin's self-repair ability. However, when cysteine ​​is used alone, its stability in the skin is poor and it is difficult to exert a long-term anti-aging effect, which limits its effective application in skin care products.

[0005] At present, although some technical means for improving the stability of active ingredients have been applied in skin care products, such as microencapsulation of peptide ingredients and liposome encapsulation, which can improve the stability and permeability of active ingredients to a certain extent, the following problems still exist: (1) Most technologies can only maintain the stability of active ingredients in the short term and it is difficult to ensure the medium- and long-term stability of active ingredients; (2) It is impossible to improve the long-term efficacy of active ingredients, and active ingredients are easily lost, resulting in limited skin care effects; (3) The targeted release of active ingredients at the site of inflammation is poor, and the anti-inflammatory effect is limited; (4) Generally, it is based on a single active ingredient, and it is difficult to effectively combine multiple ingredients with synergistic effects, and the stability and sustained-release effect of the composition are not well controlled.

[0006] In addition, studies have found that inflammatory disorders are recognized as a sign of aging and disease susceptibility, and inflammation plays a key role in regulating physiological aging. When the degree of inflammation exceeds the clearance capacity of macrophages, macrophages will also begin to secrete proinflammatory factors and reactive oxygen species, accelerating skin inflammation and damage. Proinflammatory factors are a type of aging-related secretory phenotype secreted by cells. After being absorbed by neighboring cells, they will stimulate aging-related signaling pathways and accelerate aging. Therefore, chronic inflammation will not only cause diseases in the body, but also accelerate skin aging. For example: inflammation affects the skin barrier, increases epidermal water loss, and affects the growth of keratinocytes, making it more difficult to recover after the barrier is damaged; inflammation decomposes the extracellular matrix, causing skin collapse, while inhibiting collagen synthesis, making the skin loose and wrinkled; inflammation can lead to increased tyrosinase activity and aggravate the production of melanin. Therefore, it is of great significance to develop anti-inflammatory aging skin care products. Summary of the invention

[0007] In view of the above-mentioned deficiencies, the present invention provides an anti-inflammatory aging composition, a preparation method thereof and an application thereof in skin care products. The active ingredients sea cucumber oligopeptide and cysteine ​​in the composition can synergistically enhance their effectiveness through a cross-linking reaction and the composition has good stability, and can especially target and inhibit skin inflammatory aging.

[0008] The present invention provides an anti-inflammatory aging composition, comprising sea cucumber oligopeptide, cysteine ​​and beta-cyclodextrin. The sea cucumber oligopeptide and the cysteine ​​are cross-linked through the beta-cyclodextrin to form the anti-inflammatory aging composition.

[0009] In some of the embodiments, the mass ratio of the sea cucumber oligopeptide, the cysteine ​​and the β-cyclodextrin is (0.01-2):(0.01-2.5):(0.5-2).

[0010] In some of the embodiments, the mass ratio of the sea cucumber oligopeptide, the cysteine ​​and the β-cyclodextrin is (0.8-2):(0.5-1.5):(0.8-2).

[0011] In some of the embodiments, the mass ratio of the sea cucumber oligopeptide, the cysteine ​​and the β-cyclodextrin is 1:1:1.

[0012] In some of the embodiments, the molecular weight of the sea cucumber oligopeptide is 600-2000 Da.

[0013] The present invention also provides a method for preparing the anti-inflammatory aging composition described in any one of the above technical solutions, comprising the following steps:

[0014] dissolving the sea cucumber oligopeptide in water to obtain a sea cucumber oligopeptide solution;

[0015] dissolving cysteine ​​in the sea cucumber oligopeptide solution to obtain an intermediate composition solution;

[0016] dissolving β-cyclodextrin in water to obtain a β-cyclodextrin solution;

[0017] The beta-cyclodextrin solution is slowly added to the intermediate composition solution, and the mixture is stirred to perform a cross-linking reaction to obtain an anti-inflammatory aging composition.

[0018] In some embodiments, the reaction temperature of the cross-linking reaction is 30-50° C., and the reaction time is 4-8 hours.

[0019] The present invention further provides use of the anti-inflammatory aging composition described in any one of the above technical solutions in the preparation of skin care products.

[0020] In some embodiments, the skin care product includes any one of toner, lotion, face cream, eye cream and essence.

[0021] In some of the embodiments, in the anti-inflammatory aging composition, the concentration of the sea cucumber oligopeptide is 0.01-2 mg / mL, the concentration of cysteine ​​is 0.01-2.5 mg / mL, and the concentration of β-cyclodextrin is 0.5-2 mg / mL; in the skin care product, the added amount of the anti-inflammatory aging composition is 0.05% to 1% of the quality of the skin care product.

[0022] Compared with the prior art, the advantages and beneficial effects of the present invention are:

[0023] 1. The anti-inflammatory aging composition provided by the present invention uses β-cyclodextrin as a molecular cross-linking agent to cross-link sea cucumber oligopeptide and cysteine ​​to form a low-molecular complex, which not only improves the stability of the two active ingredients, solves the problem of poor stability when sea cucumber oligopeptide or cysteine ​​is used as a single ingredient, but also improves the antioxidant and anti-aging effects through the synergistic effect of the two.

[0024] 2. The anti-inflammatory aging composition provided by the present invention has active ingredients that can provide long-lasting skin care effects, and has targeted inflammation inhibition effects, and can effectively improve the inflammatory state;

[0025] 3. The preparation method of the anti-inflammatory aging composition provided by the present invention uses water as a solvent, which is convenient for adding to skin care products, and the preparation steps are simple, without the need for complex equipment and cumbersome process flow, thus reducing production costs and energy consumption, being conducive to large-scale industrial production, and having good economy and sustainability;

[0026] 4. The anti-inflammatory aging composition provided by the present invention has good market potential and application prospects when used in the preparation of skin care products. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The stability test results of the compositions prepared in Example 1 and Comparative Example 1 of the present invention are shown in FIG. 1 , wherein (a) is the stability test result of sea cucumber oligopeptide, and (b) is the stability test result of cysteine;

[0028] Figure 2 The test results of type I collagen level detection of the anti-inflammatory aging composition prepared in Example 1 of the present invention and sea cucumber oligopeptides, cysteine ​​and β-cyclodextrin;

[0029] Figure 3 The DPPH free radical scavenging test results of the anti-inflammatory aging composition prepared in Example 1 of the present invention and sea cucumber oligopeptides, cysteine ​​and β-cyclodextrin;

[0030] Figure 4 The results of the targeted release test of the anti-inflammatory aging composition prepared in Example 1 of the present invention;

[0031] Figure 5 The test results of type I collagen level detection of the compositions prepared in Examples 1-3 and Comparative Examples 2-7 of the present invention;

[0032] Figure 6 The DPPH free radical scavenging test results of the compositions prepared in Examples 1-3 and Comparative Examples 2-7 of the present invention;

[0033] Figure 7 These are the test results for detecting the transcription levels of inflammatory factor genes of the compositions prepared in Examples 1-3 and Comparative Examples 2-7 of the present invention. DETAILED DESCRIPTION

[0034] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0035] An embodiment of the present invention provides an anti-inflammatory aging composition, comprising sea cucumber oligopeptide, cysteine ​​and β-cyclodextrin, wherein the sea cucumber oligopeptide and the cysteine ​​are cross-linked through the β-cyclodextrin to form the anti-inflammatory aging composition.

[0036] The above anti-inflammatory aging composition uses β-cyclodextrin as a molecular cross-linking agent to cross-link sea cucumber oligopeptide with cysteine ​​to form a low molecular weight complex, thereby maintaining the unique activity of the two active ingredients, and the structure of the formed low molecular weight complex helps the active ingredients enter the basal layer of the skin and enhance the skin repair effect. At the same time, the above anti-inflammatory aging composition uses β-cyclodextrin to cross-link sea cucumber oligopeptide with cysteine ​​to form a three-dimensional supramolecular network, thereby improving the stability of the two active ingredients, specifically: (1) the hydrophobic cavity of β-cyclodextrin interacts with the hydrophobic group of the sea cucumber oligopeptide to form a host-guest inclusion complex, thereby achieving the embedding of the active center of the sea cucumber oligopeptide, reducing the exposure of the protease attack site, and improving the in vitro digestion stability of the sea cucumber oligopeptide; (2) the hydroxyl group of β-cyclodextrin forms an intermolecular hydrogen bond network with the carboxyl group of cysteine, interfering with the orderly arrangement of the peptide chain, so that the anti-inflammatory aging composition remains amorphous during long-term storage, avoiding activity loss and improving stability. Furthermore, the above anti-inflammatory aging composition achieves synergistic effects of two active ingredients through the combination of sea cucumber oligopeptides and cysteine, specifically: (1) the antioxidant peptide segment of the sea cucumber oligopeptide forms an electron relay transfer chain with the thiol group of cysteine, and the synergistic effect of the two improves the free radical scavenging efficiency; (2) the sea cucumber oligopeptide promotes collagen synthesis and thus increases skin elasticity, which, combined with the antioxidant effect of cysteine, effectively improves the structure and function of the skin and delays the aging process. In addition, the sea cucumber oligopeptide segment of the above anti-inflammatory aging composition is degraded under the action of matrix metalloproteinases (MMPs) that occur with skin inflammation, so that the three-dimensional supramolecular network of the anti-inflammatory aging composition dissociates in the inflammatory microenvironment, achieving targeted release of active ingredients and being able to effectively improve the inflammatory state.

[0037] In a preferred embodiment, the mass ratio of the sea cucumber oligopeptide, the cysteine ​​and the β-cyclodextrin is (0.01-2):(0.01-2.5):(0.5-2). This preferred embodiment further defines the preferred ratio of sea cucumber oligopeptide, cysteine ​​and β-cyclodextrin. The anti-inflammatory aging composition obtained under this preferred ratio has good antioxidant and anti-aging effects, and its active ingredients can provide long-lasting skin care effects and have the effect of targeted inhibition of inflammation.

[0038] In a preferred embodiment, the mass ratio of the sea cucumber oligopeptide, the cysteine ​​and the β-cyclodextrin is (0.8-2):(0.5-1.5):(0.8-2). This preferred embodiment further defines the preferred ratio of sea cucumber oligopeptide, cysteine ​​and β-cyclodextrin, and the anti-inflammatory aging composition obtained under this preferred ratio is more suitable for preparing skin care products.

[0039] In a preferred embodiment, the mass ratio of the sea cucumber oligopeptide, the cysteine ​​and the β-cyclodextrin is 1:1:1. This preferred embodiment further defines the optimal ratio of sea cucumber oligopeptide, cysteine ​​and β-cyclodextrin, and the anti-inflammatory aging composition obtained under the optimal ratio has more significant anti-aging ability and antioxidant activity.

[0040] In a preferred embodiment, the molecular weight of the sea cucumber oligopeptide is 600 to 2000 Da. This preferred embodiment further limits the preferred range of the molecular weight of the sea cucumber oligopeptide. Sea cucumber oligopeptides with too large molecular weight are not conducive to forming low molecular weight complexes after cross-linking, and are not conducive to skin absorption, while sea cucumber oligopeptides with too small molecular weight are not conducive to cross-linking to form a three-dimensional supramolecular network structure.

[0041] It should be noted that the preparation method of sea cucumber oligopeptide is as follows:

[0042] (1) taking fresh sea cucumber, cleaning and removing sand, cutting into small pieces, weighing, adding ultrapure water in a ratio of 1:4 (m / w), homogenizing, and preparing a sea cucumber homogenate;

[0043] (2) adjusting the pH of the sea cucumber homogenate to neutral, adding flavor protease at a ratio of 4200 U / g solid content, performing enzymatic hydrolysis at 45° C. for 6 h, inactivating the enzyme, and obtaining a sea cucumber enzymatic hydrolyzate;

[0044] (3) The pH of the sea cucumber hydrolysate was adjusted to 6, and a mixed carbohydrase (the mass ratio of cellulase to pectinase was 1:2) was added at 0.12%, and the enzymatic hydrolysis was carried out at 50° C. for 3.6 h, and the enzyme was inactivated for 20 min. The pH was adjusted to neutral, and the solution was filtered to obtain a sea cucumber peptide hydrolysate;

[0045] (4) adding 1.2% activated carbon to the sea cucumber peptide hydrolysate, decolorizing at 70°C for 50 min, refrigerating centrifuging at 10000 r / min for 10 min, collecting the supernatant, and ultrafiltration through a 0.45 μm membrane to obtain a sea cucumber decolorizing solution;

[0046] (5) using an organic membrane system to separate, purify and concentrate the sea cucumber decolorization solution, wherein a 2000Da organic membrane is used, the temperature is controlled at 10-20°C, the pressure is 0.25MPa, and the flow rate is 1.6mL / s; collecting the concentrated solution to obtain the final enzymatic hydrolyzate of the sea cucumber oligopeptide;

[0047] (6) The final enzymatic hydrolyzate of the sea cucumber oligopeptide is frozen in a plate, and further vacuum-freeze-dried to obtain refined sea cucumber oligopeptide.

[0048] The molecular weight of the sea cucumber oligopeptide prepared by the preparation method is between 600 and 2000 Da.

[0049] It should also be noted that cysteine ​​is a conditionally essential amino acid for the human body. Its chemical name is 2-amino-3-mercaptopropionic acid and its chemical formula is C 3 H 7 NO 2 S, the purity of cysteine ​​used in the anti-inflammatory aging composition must be above 99%, and the structural formula of cysteine ​​is as follows:

[0050]

[0051] It should be further explained that β-cyclodextrin is a conical ring with a hydrophilic outer edge and a hydrophobic inner cavity. Its chemical formula is C 42 H 70 O 35 The purity of β-cyclodextrin used in the anti-inflammatory aging composition must be above 99%. The structural formula of β-cyclodextrin is as follows:

[0052]

[0053] The present invention also provides a method for preparing an anti-inflammatory aging composition, comprising the following steps:

[0054] S1, dissolving the sea cucumber oligopeptide in water to obtain a sea cucumber oligopeptide solution;

[0055] S2, dissolving cysteine ​​in the sea cucumber oligopeptide solution to obtain an intermediate composition solution;

[0056] S3, dissolving β-cyclodextrin in water to obtain a β-cyclodextrin solution;

[0057] S4. Slowly add the β-cyclodextrin solution to the intermediate composition solution, stir to carry out a cross-linking reaction, and obtain an anti-inflammatory aging composition.

[0058] The preparation method of the anti-inflammatory aging composition uses water as a solvent, which is convenient for adding to skin care products, and the preparation steps are simple, without the need for complex equipment and cumbersome process flow, thus reducing production costs and energy consumption, being conducive to large-scale industrial production, and having good economy and sustainability.

[0059] In a preferred embodiment, in step S1, according to the solubility of the sea cucumber oligopeptide, it is preferably dissolved in pure water at 15 to 20° C. and ultrasonically dissolved for 25 to 35 minutes.

[0060] In a preferred embodiment, in step S2, according to the solubility of cysteine, it is preferably dissolved at 25-30° C. and ultrasonically dissolved for 25-35 min.

[0061] In a preferred embodiment, in step S3, according to the solubility of β-cyclodextrin, it is preferably dissolved in pure water at 30-50° C. and stirred for 1 hour.

[0062] In a preferred embodiment, in step S4, the reaction temperature of the cross-linking reaction is 30-50°C and the reaction time is 4-8 hours. This preferred embodiment further defines the preferred reaction temperature and time of the cross-linking reaction, which is conducive to ensuring the cross-linking to form a three-dimensional supramolecular network.

[0063] The embodiments of the present invention further provide the use of the anti-inflammatory aging composition in the preparation of skin care products. It should be noted that the anti-inflammatory aging composition provided in the embodiments of the present invention is not only used to prepare anti-aging and repair skin care products, but can also be extended to other skin care products, such as moisturizing, anti-oxidation, whitening, protection and other fields, and has great market potential and application prospects.

[0064] In a preferred embodiment, the skin care products include any one of toner, lotion, face cream, eye cream and essence, for example, anti-aging repair cream, antioxidant essence, soothing repair night cream, whitening essence and deep moisturizing essence.

[0065] In a preferred embodiment, in the anti-inflammatory aging composition, the concentration of the sea cucumber oligopeptide is 0.01-2 mg / mL, the concentration of cysteine ​​is 0.01-2.5 mg / mL, and the concentration of β-cyclodextrin is 0.5-2 mg / mL; in the skin care product, the amount of the anti-inflammatory aging composition added is 0.05%-1% of the quality of the skin care product. This preferred embodiment further defines the preferred range of the amount of the anti-inflammatory aging composition added to the skin care product, and at this preferred amount, a significant effect can be obtained.

[0066] In order to more clearly and in detail introduce the anti-inflammatory aging composition and its preparation method and its application in skin care products provided by the embodiments of the present invention, the following will be described in conjunction with specific examples. It should be noted that the preparation method of the sea cucumber oligopeptide used in the following embodiments is as follows:

[0067] (1) taking fresh sea cucumber, cleaning and removing sand, cutting into small pieces, weighing, adding ultrapure water in a ratio of 1:4 (m / w), homogenizing, and preparing a sea cucumber homogenate;

[0068] (2) adjusting the pH of the sea cucumber homogenate to neutral, adding flavor protease at a ratio of 4200 U / g solid content, performing enzymatic hydrolysis at 45° C. for 6 h, inactivating the enzyme, and obtaining a sea cucumber enzymatic hydrolyzate;

[0069] (3) The pH of the sea cucumber hydrolysate was adjusted to 6, and a mixed carbohydrase (the mass ratio of cellulase to pectinase was 1:2) was added at 0.12%, and the enzymatic hydrolysis was carried out at 50° C. for 3.6 h, and the enzyme was inactivated for 20 min. The pH was adjusted to neutral, and the solution was filtered to obtain a sea cucumber peptide hydrolysate;

[0070] (4) adding 1.2% activated carbon to the sea cucumber peptide hydrolysate, decolorizing at 70°C for 50 min, refrigerating centrifuging at 10000 r / min for 10 min, collecting the supernatant, and ultrafiltration through a 0.45 μm membrane to obtain a sea cucumber decolorizing solution;

[0071] (5) using an organic membrane system to separate, purify and concentrate the sea cucumber decolorization solution, wherein a 2000Da organic membrane is used, the temperature is controlled at 10-20°C, the pressure is 0.25MPa, and the flow rate is 1.6mL / s; collecting the concentrated solution to obtain the final enzymatic hydrolyzate of the sea cucumber oligopeptide;

[0072] (6) The final enzymatic hydrolyzate of the sea cucumber oligopeptide is frozen in a plate, and further freeze-dried in a vacuum to obtain the refined sea cucumber oligopeptide. The molecular weight of the sea cucumber oligopeptide is between 600 and 2000 Da.

[0073] 1. Specific Examples of Anti-Inflammatory Aging Composition and Preparation Method thereof

[0074] Example 1

[0075] An anti-inflammatory aging composition comprises 1g of sea cucumber oligopeptide, 1g of cysteine ​​(with a purity greater than 99%) and 1g of beta-cyclodextrin (with a purity greater than 99%), wherein the sea cucumber oligopeptide and cysteine ​​are cross-linked by beta-cyclodextrin to form the anti-inflammatory aging composition.

[0076] The method for preparing the anti-inflammatory aging composition comprises the following steps:

[0077] (1) dissolving the sea cucumber oligopeptide in 600 mL of 20° C. pure water, and ultrasonically dissolving for 30 min to obtain a sea cucumber oligopeptide solution;

[0078] (2) heating the sea cucumber oligopeptide solution to 25° C., adding cysteine, and dissolving by ultrasonication for 30 min to obtain an intermediate composition solution;

[0079] (3) dissolving β-cyclodextrin in 400 mL of 40° C. pure water, stirring and dissolving at a constant temperature for 1 h to obtain a β-cyclodextrin solution;

[0080] (4) The intermediate composition solution is heated to 40° C., the β-cyclodextrin solution is slowly added thereto, and the mixture is stirred and reacted at a constant temperature for 6 hours to obtain an anti-inflammatory aging composition.

[0081] Example 2

[0082] The difference between this embodiment and embodiment 1 is that the dosage of each component is different. In this embodiment, the amount of sea cucumber oligopeptide is 0.01 g, the amount of cysteine ​​is 0.01 g, and the amount of β-cyclodextrin is 0.5 g.

[0083] Example 3

[0084] The difference between this embodiment and embodiment 1 is that the dosage of each component is different. In this embodiment, the amount of sea cucumber oligopeptide is 2 g, the amount of cysteine ​​is 2.5 g, and the amount of β-cyclodextrin is 2 g.

[0085] Comparative Example 1

[0086] The difference between this embodiment and embodiment 1 is that β-cyclodextrin is not added. The preparation method of the composition provided in this embodiment is as follows:

[0087] (1) dissolving the sea cucumber oligopeptide in 600 mL of 20° C. pure water, and ultrasonically dissolving for 30 min to obtain a sea cucumber oligopeptide solution;

[0088] (2) heating the above sea cucumber oligopeptide solution to 25° C., adding cysteine, and dissolving by ultrasonication for 30 min to uniformly mix the cysteine ​​and the sea cucumber oligopeptide to obtain a mixture solution;

[0089] (3) The mixture solution was heated to 400 mL at 40° C. and stirred for reaction at constant temperature for 6 h to obtain a composition of sea cucumber oligopeptide and cysteine.

[0090] Comparative Example 2

[0091] The difference between this embodiment and embodiment 1 is that the dosage of each component is different. In this embodiment, the amount of sea cucumber oligopeptide is 0.001 g, the amount of cysteine ​​is 0.01 g, and the amount of β-cyclodextrin is 0.5 g.

[0092] Comparative Example 3

[0093] The difference between this embodiment and embodiment 1 is that the dosage of each component is different. In this embodiment, the amount of sea cucumber oligopeptide is 2.5 g, the amount of cysteine ​​is 2.5 g, and the amount of β-cyclodextrin is 2 g.

[0094] Comparative Example 4

[0095] The difference between this embodiment and embodiment 1 is that the dosage of each component is different. In this embodiment, the amount of sea cucumber oligopeptide is 0.01 g, the amount of cysteine ​​is 0.001 g, and the amount of β-cyclodextrin is 0.5 g.

[0096] Comparative Example 5

[0097] The difference between this embodiment and embodiment 1 is that the dosage of each component is different. In this embodiment, the amount of sea cucumber oligopeptide is 2 g, the amount of cysteine ​​is 3 g, and the amount of β-cyclodextrin is 2 g.

[0098] Comparative Example 6

[0099] The difference between this embodiment and embodiment 1 is that the dosage of each component is different. In this embodiment, the amount of sea cucumber oligopeptide is 0.01 g, the amount of cysteine ​​is 0.01 g, and the amount of β-cyclodextrin is 0.1 g.

[0100] Comparative Example 7

[0101] The difference between this embodiment and embodiment 1 is that the dosage of each component is different. In this embodiment, the amount of sea cucumber oligopeptide is 2 g, the amount of cysteine ​​is 2.5 g, and the amount of β-cyclodextrin is 2.5 g.

[0102] Performance Testing

[0103] 1. Stability test

[0104] For the compositions prepared in Example 1 and Comparative Example 1, the sea cucumber oligopeptide content and cysteine ​​content in the compositions were detected at 25° C. every 24 hours for 3 consecutive days.

[0105] The method for detecting the content of sea cucumber oligopeptides is based on the manufacturer's instructions of the BeyoBCA peptide concentration determination kit. The specific method is as follows:

[0106] (1) Prepare 0, 15.625, 31.25, 62.5, 125, 250, 500, and 1000 μg / mL peptide standards, and mix thoroughly during each dilution;

[0107] (2) Prepare BeyoBCA working solution: Each reaction requires 180 μl of BeyoBCA working solution. Prepare an appropriate amount of BeyoBCA working solution according to the volume ratio of ReagentA: ReagentB: ReagentC = 50:48:2 (i.e., 50 volumes of ReagentA, 48 volumes of ReagentB, and 2 volumes of ReagentC). Mix thoroughly. The BeyoBCA working solution is stable at room temperature within 30 minutes.

[0108] (3) Add 20 μL of peptide standards of different concentrations and the samples to be tested to the standard wells of a 96-well plate, add 180 μL of working solution to each well, mix for 1 minute, and place at 37°C for 15 minutes;

[0109] (4) Use an enzyme-labeled instrument to measure the absorbance of the standard and sample at A480 wavelength, where the absorbance measurement value of the standard and sample must be subtracted from the absorbance measurement value of the 0 μg / mL blank standard. Draw a standard curve based on the absorbance of the standard, and calculate the polypeptide concentration of the sample well based on the standard curve, which is the sea cucumber oligopeptide content.

[0110] The results of the stability test of sea cucumber oligopeptides are as follows Figure 1 As shown in a, the results show that compared with the composition without adding β-cyclodextrin, the sea cucumber oligopeptide in the anti-inflammatory aging composition prepared by adding β-cyclodextrin in Example 1 of the present invention has higher stability.

[0111] The method for detecting cysteine ​​content is based on the manufacturer's instructions of the cysteine ​​(Cys) content detection kit. The specific method is as follows:

[0112] (1) Take 0.2 mL of sample, add 0.3 mL of extract, mix thoroughly, centrifuge at 11000 rpm and 4°C for 10 min, and take the supernatant for testing;

[0113] (2) Add the standard to 0.826 mL of pure water to obtain a 100 μmol / mL standard solution, which is then diluted with water to obtain standard solutions with concentrations of 20, 10, 5, 2.5, 1.25, and 0.625 μmol / mL for drawing a standard curve;

[0114] (3) Add the sample to be tested, pure water, reagent 1 and reagent 2 to a 96-well plate in sequence, mix well and let stand at room temperature for 15 min, measure the absorbance at 600 nm, and calculate the cysteine ​​content according to the standard curve.

[0115] The results of the cysteine ​​stability test are as follows Figure 1 As shown in b, the results show that compared with the composition provided in Comparative Example 1 without adding β-cyclodextrin, the cysteine ​​in the anti-inflammatory aging composition prepared by adding β-cyclodextrin in Example 1 of the present invention has higher stability.

[0116] 2. Activity test of the composition

[0117] 2.1. Anti-aging ability test

[0118] The sea cucumber oligopeptide, cysteine, β-cyclodextrin and the anti-inflammatory aging composition prepared in Example 1 were subjected to a test for detecting the type I collagen level in zebrafish embryos to evaluate the anti-aging ability.

[0119] The test groups are as follows:

[0120] Blank group: fish culture water;

[0121] H 2 O 2 Model group: 20 μmol / L H 2 O 2 ;

[0122] The treatment groups included: H 2 O 2 and sea cucumber oligopeptide group (H 2 O 2 The concentration of 20 μmol / L and the concentration of sea cucumber oligopeptide were 5 μg / mL, and the H 2 O 2 and cysteine ​​group (H 2 O 2The concentration of 20 μmol / L and cysteine ​​concentration of 5 μg / mL) were prepared with H 2 O 2 and β-cyclodextrin group (H 2 O 2 The concentration of 20 μmol / L and 5 μg / mL of β-cyclodextrin were prepared with H 2 O 2 and Example 1 Composition Group (H 2 O 2 The concentration was 20 μmol / L, and the composition of Example 1 was diluted 200 times).

[0123] The zebrafish embryos exposed to 96 hpf in the above experimental groups were collected by grinding and centrifugation, and the type I collagen content was detected.

[0124] The test was performed according to the manufacturer's instructions of the type I collagen ELISA test kit. The specific method was as follows: take out the ELISA plate, add a wash solution diluted to 1 times the concentration (350 μL / well) to soak, let it stand for 30 seconds, discard the liquid in the wells, and pat the ELISA plate dry; add 100 μL of the standard and the sample to be tested to each well, and incubate at room temperature for 2 hours; discard the liquid in the wells, add the detection antibody, and incubate at room temperature for 2 hours; discard the liquid in the wells again, add the enzyme conjugate, and incubate at room temperature for 20 minutes; after the incubation, discard the liquid in the wells, add the substrate solution, and incubate at room temperature in the dark for 20 minutes; finally, add the stop solution and detect the absorbance value.

[0125] The results of the anti-aging test of sea cucumber oligopeptides, cysteine, β-cyclodextrin and Example 1 are as follows Figure 2 As shown, it can be seen that sea cucumber oligopeptides, cysteine ​​and the anti-inflammatory aging composition provided in Example 1 of the present invention all significantly increased the content of type I collagen, and compared with sea cucumber oligopeptides and cysteine, the anti-inflammatory aging composition increased the content of type I collagen to a greater extent, which shows that in the anti-inflammatory aging composition provided in Example 1 of the present invention, sea cucumber oligopeptides and cysteine ​​have a synergistic effect.

[0126] 2.2 Antioxidant activity test

[0127] The sea cucumber oligopeptide, cysteine, β-cyclodextrin and the anti-inflammatory aging composition prepared in Example 1 were subjected to a DPPH free radical scavenging test to evaluate the antioxidant activity.

[0128] The test groups are as follows:

[0129] Blank group: no sample was added;

[0130] The treatment groups included: a sea cucumber oligopeptide group (sea cucumber oligopeptide concentration was 5 μg / mL), a cysteine ​​group (cysteine ​​concentration was 5 μg / mL), a β-cyclodextrin group (β-cyclodextrin concentration was 5 μg / mL) and an Example 1 group (the composition of Example 1 was diluted 200 times).

[0131] Take 0.1 mL of the sample from the above test groups and add 3.9 mL of DPPH solution (concentration of 6×10 -5 mol / L), and placed in the dark at room temperature for 2 h, and its absorbance was measured at a wavelength of 515 nm.

[0132] DPPH free radical scavenging rate = (A 0 -A b ) / A 0 ×100%

[0133] A 0 Indicates the absorbance value of DPPH solution; A b Represents the absorbance values ​​of DPPH solutions of different sample groups.

[0134] The results of the antioxidant activity test of sea cucumber oligopeptides, cysteine, β-cyclodextrin and Example 1 are as follows Figure 3 As shown, it can be seen that sea cucumber oligopeptides, cysteine ​​and the anti-inflammatory aging composition provided in Example 1 of the present invention all have significant free radical scavenging capabilities, and the free radical scavenging effect of the anti-inflammatory aging composition provided in Example 1 of the present invention is the most significant, and its free radical scavenging level is better than the effect of sea cucumber oligopeptides and cysteine ​​used alone. This shows that in the anti-inflammatory aging composition provided in Example 1 of the present invention, sea cucumber oligopeptides and cysteine ​​have a synergistic effect.

[0135] 2.3 Targeted release ability test

[0136] The anti-inflammatory aging composition prepared in Example 1 was subjected to a targeted release test of active ingredients under inflammatory conditions based on zebrafish to evaluate the targeted release capability.

[0137] The test groups are as follows:

[0138] Blank group: fish culture water;

[0139] LPS model group: 20 μg / mL LPS prepared in fish culture water;

[0140] The drug treatment groups included: MMP group (MMP concentration was 0.1 μmol / L), LPS and MMP group (LPS concentration was 20 μg / mL, MMP concentration was 0.1 μmol / L), LPS and Example 1 group (LPS concentration was 20 μg / mL, and the composition of Example 1 was diluted 200 times), and LPS, Example 1 and MMP mixed group (LPS concentration was 20 μg / mL, MMP concentration was 0.1 μmol / L, and the composition of Example 1 was diluted 200 times).

[0141] The zebrafish embryos were incubated to 96 hpf using the above experimental groups, and the transcription levels of inflammatory factor genes were detected.

[0142] Primer Blast software was used to design qPCR primers for the detection of inflammatory factor gene transcription levels, and all primers followed the primer design principles. RNA was extracted from zebrafish embryos exposed to 96 hpf in the above experimental groups to obtain total RNA; the obtained RNA was reverse transcribed to obtain cDNA; real-time fluorescence quantitative PCR was performed according to the manufacturer's instructions, and 2 was used according to the CT value. －ΔΔct The relative expression levels of genes were statistically analyzed by this method.

[0143] It should be noted that MMP is usually accompanied by the appearance of skin inflammation. Therefore, the purpose of this experiment is to verify the inhibitory effect of Example 1 in the presence of MMP. If the inhibitory effect of Example 1 with the addition of MMP is better than that of Example 1 without the addition of MMP, it indicates that Example 1 can target and identify skin inflammation accompanied by MMP and can perform better repair at this site. The test results of the targeted release ability of the anti-inflammatory aging composition prepared in Example 1 are as follows: Figure 4 As shown, it can be seen that compared with the Example 1 group without adding MMP, the expression levels of inflammatory factors in Example 1 and the MMP mixed group with added MMP were significantly reduced, which shows that the anti-inflammatory aging composition provided in Example 1 of the present invention can target and identify skin inflammation accompanied by MMP, and can perform better repair at this site.

[0144] 3. Comparative test of the activity of the composition under different ratios

[0145] 3.1. Cross-linking status comparison test

[0146] The solution states of the compositions of Examples 1-3 and Comparative Examples 2-7 after standing for 24 hours were observed. The results are summarized in Table 1.

[0147] Table 1 Comparison of crosslinking states of compositions at different ratios

[0148]

[0149] 3.2. Anti-aging ability comparison test

[0150] The compositions prepared in Examples 1-3 and Comparative Examples 2-7 were subjected to a test for detecting type I collagen levels in zebrafish embryos to evaluate their anti-aging abilities.

[0151] The test groups are as follows:

[0152] Blank group: fish culture water;

[0153] H 2 O 2 Model group: 20 μmol / L H 2 O 2 ;

[0154] The treatment groups included: H 2 O 2 and Example 1 group, H 2 O 2 and Example 2 Group, H 2 O 2 and Example 3 Group, H 2 O 2 And the comparative example 2 group, H 2 O 2 And the comparative example 3 group, H 2 O 2 And the comparative example 4 group, H 2 O 2 And the comparative example 5 group, H 2 O 2 And the comparative example 6 group, H 2 O 2 and 7 groups of comparative examples (in each treatment group, H 2 O 2 The concentration is 20 μmol / L, the composition is diluted 200 times)

[0155] The test method for detecting type I collagen level is the same as the test method for detecting type I collagen level used in the above-mentioned composition activity test. The test results are shown in FIG. Figure 5 As shown, it can be seen that compared with the compositions prepared in Comparative Examples 2-7, the anti-inflammatory aging compositions prepared in Examples 1-3 significantly increased the content of type I collagen.

[0156] 3.3 Antioxidant activity test

[0157] The compositions prepared in Examples 1-3 and Comparative Examples 2-7 were subjected to a DPPH free radical scavenging test to evaluate their antioxidant activity.

[0158] The test groups are as follows:

[0159] Blank group: no sample was added;

[0160] The treatment groups include: Example 1 group, Example 2 group, Example 3 group, Comparative Example 2 group, Comparative Example 3 group, Comparative Example 4 group, Comparative Example 5 group, Comparative Example 6 group and Comparative Example 7 group (in each treatment group, the composition was diluted 200 times).

[0161] The DPPH free radical scavenging test method is the same as the DPPH free radical scavenging test method used in the above-mentioned composition activity test. The test results are as follows: Figure 6 As shown, it can be seen that compared with the compositions prepared in Comparative Examples 2-7, the anti-inflammatory aging compositions prepared in Examples 1-3 have significant free radical scavenging capabilities.

[0162] 3.4. Comparative test of anti-inflammatory ability

[0163] The compositions prepared in Examples 1-3 and Comparative Examples 2-7 were subjected to inflammatory factor gene transcription level detection based on zebrafish to evaluate the anti-inflammatory ability.

[0164] The test groups are as follows:

[0165] Blank group: fish culture water;

[0166] LPS model group: 20 μg / mL LPS prepared in fish culture water;

[0167] The drug treatment groups included: LPS and Example 1 group, LPS and Example 2 group, LPS and Example 3 group, LPS and Comparative Example 2 group, LPS and Comparative Example 3 group, LPS and Comparative Example 4 group, LPS and Comparative Example 5 group, LPS and Comparative Example 6 group, LPS and Comparative Example 7 group (in each treatment group, the concentration of LPS was 20 μg / mL, and the composition was diluted 200 times).

[0168] The method for detecting the transcription level of inflammatory factor genes is the same as the method for detecting the transcription level of inflammatory factor genes used in the above-mentioned composition activity test. The test results are compared as follows: Figure 7 As shown, it can be seen that compared with the compositions prepared in Comparative Examples 2-7, the anti-inflammatory aging compositions prepared in Examples 1-3 have significant anti-inflammatory effects.

[0169] 2. Specific examples of the application of the anti-inflammatory aging composition in skin care products

[0170] Example 4: Anti-aging repair cream

[0171] The anti-inflammatory aging composition comprises: 2g sea cucumber oligopeptide, 0.5g cysteine ​​(purity greater than 99%) and 2g beta-cyclodextrin (purity greater than 99%), wherein the sea cucumber oligopeptide and cysteine ​​are cross-linked by beta-cyclodextrin to form the anti-inflammatory aging composition.

[0172] The preparation method of the anti-inflammatory aging composition is the same as that of Example 1, wherein the concentration of sea cucumber oligopeptide is 2 mg / mL, the concentration of cysteine ​​is 0.05 mg / mL, and the concentration of β-cyclodextrin is 2 mg / mL.

[0173] An anti-aging repairing facial cream comprises the following components in percentage by mass: 1% of the above-mentioned anti-inflammatory aging composition (used to promote collagen synthesis, enhance skin elasticity, and resist oxidation, slow down the skin aging process, and reduce free radical damage), 3% of glycerin (as a main moisturizer, with excellent hydration, helping to maintain skin moisture balance and enhancing skin barrier function), 0.5% of polyvinyl alcohol (providing a smooth texture and moisturizing effect, enhancing the comfort of the cream), 2% of squalene (having a moisturizing effect, while helping to enhance the skin barrier function and improve the gloss and softness of the skin), 15% of polysorbate-80 (as an emulsifier), 0.8% of phenoxyethanol (as a preservative), xanthan gum (as a thickener, the amount of which is adjusted according to the target viscosity state) and citric acid (as a pH regulator, the amount of which is determined according to the target pH value), and the rest is deionized water.

[0174] The preparation method of the above-mentioned anti-aging repairing cream comprises the following steps:

[0175] (1) Raw material preparation: weigh each component according to the amount of each component in the above anti-aging repair cream and set aside;

[0176] (2) Preparation of aqueous phase: Add the anti-inflammatory aging composition and glycerin to deionized water, and stir at 100-200 rpm for 15 minutes until all ingredients are completely dissolved; add xanthan gum and continue stirring at 300-400 rpm for 10 minutes until the solution is uniform and slightly viscous; slowly add citric acid dropwise while stirring to adjust the pH to 5.5; it should be noted that the aqueous phase preparation process should be kept at room temperature (20-25°C) to avoid degradation of ingredients due to excessively high temperatures;

[0177] (3) Preparation of oil phase: Add polysorbate 80 and squalene to another container, stir and heat to 60°C, and continue stirring for 10 minutes until polysorbate 80 and squalene are completely dissolved to form a transparent and uniform oil phase; it should be noted that the oil phase preparation process should be maintained at 60°C ± 5°C to avoid excessively high temperatures that may cause degradation of polysorbate 80;

[0178] (4) Emulsification: slowly add the prepared water phase to the oil phase, use an emulsifier for efficient emulsification, set the speed of the emulsifier to 1500 rpm, and emulsify for 15 minutes to ensure that the water phase and the oil phase are completely fused to form a stable emulsion; it should be noted that the viscosity and texture of the emulsion can be monitored during the emulsification process to ensure that it is uniform and without stratification; during the emulsification process, the temperature of the oil phase should be maintained at about 60°C, while the temperature of the water phase should be close to room temperature (20-25°C);

[0179] (5) Adding polyvinyl alcohol and phenoxyethanol: After emulsification is completed, slowly add polyvinyl alcohol and stir with a high shear stirrer for 15 minutes to ensure that it is fully dissolved and evenly dispersed in the emulsion; after the emulsion is stirred evenly, add phenoxyethanol and continue stirring for 10 minutes to ensure that it is completely dissolved in the emulsion;

[0180] (6) Cooling and pH adjustment: Place the mixed emulsion on a cooling rack and cool it naturally to room temperature (20-25° C.). After cooling, check the pH value of the emulsion again and use citric acid to make a fine adjustment to ensure that the final pH value is 5.5 to obtain an anti-aging repair cream.

[0181] (7) Inspection and packaging:

[0182] Product inspection: After confirming that the appearance and sensory effects of the cream meet the standards, a stability check is conducted to ensure that there is no stratification, precipitation, or bubbles;

[0183] Subpackaging: The final anti-aging repair cream is packaged into pre-sterilized containers and sealed for storage. It should be noted that the packaging environment must be kept clean during the subpackaging process to avoid product contamination;

[0184] Storage: Store in a cool and dry place, avoid direct sunlight and maintain constant room temperature.

[0185] Example 5: Antioxidant Essence

[0186] The anti-inflammatory aging composition comprises: 1g of sea cucumber oligopeptide, 1.5g of cysteine ​​(purity greater than 99%) and 1g of beta-cyclodextrin (purity greater than 99%), wherein the sea cucumber oligopeptide and cysteine ​​are cross-linked by beta-cyclodextrin to form the anti-inflammatory aging composition.

[0187] The preparation method of the anti-inflammatory aging composition is the same as that of Example 1, wherein the concentration of sea cucumber oligopeptide is 1 mg / mL, the concentration of cysteine ​​is 1.5 mg / mL, and the concentration of β-cyclodextrin is 1 mg / mL.

[0188] An antioxidant essence comprises the following components in percentage by mass: 0.1% of the above-mentioned anti-inflammatory aging composition (for promoting collagen synthesis, enhancing skin elasticity, and anti-oxidation, slowing down the skin aging process, and reducing free radical damage), 1% hyaluronic acid (with a strong moisturizing effect, capable of penetrating into the bottom layer of the skin, enhancing skin elasticity, and improving the firmness and moisture of the skin), 0.5% vitamin E (as a powerful antioxidant, providing additional antioxidant protection and enhancing the skin's ability to resist aging), 0.8% phenoxyethanol (as a preservative) and citric acid (as a pH adjuster, the amount of which is determined according to the target pH value), and the remainder is deionized water.

[0189] The method for preparing the above-mentioned antioxidant essence comprises the following steps:

[0190] (1) Raw material preparation: weigh each component according to the amount of each component in the above antioxidant essence and set aside;

[0191] (2) Preparation of aqueous phase: adding the anti-inflammatory aging composition to deionized water, and stirring at 200 rpm for 10 min using a stirrer until the components are evenly dispersed;

[0192] (3) Adding hyaluronic acid and vitamin E: Add hyaluronic acid to the aqueous phase and stir at 100 rpm for 10 min to ensure that the hyaluronic acid is fully dissolved and evenly dispersed; further add vitamin E and continue to stir evenly to ensure that it is completely dissolved and fully mixed with other ingredients;

[0193] (4) Ultrasonic treatment: The mixture is granulated using ultrasonic treatment technology to ensure that the particle size is between 100-300 nm to enhance permeability and bioavailability, wherein the ultrasonic power is set to 100 W and the treatment time is 10 min. It should be noted that during the ultrasonic treatment process, the temperature of the mixture is controlled at 25°C ± 2°C to avoid degradation of the components due to excessively high temperature.

[0194] (5) Adding phenoxyethanol: After the ultrasonic treatment, add phenoxyethanol and stir for 10 min to ensure that the phenoxyethanol is completely dissolved. Check the stability to ensure that there is no precipitation or stratification.

[0195] (6) pH adjustment: Use citric acid to adjust the pH of the product to 5.5 to obtain an antioxidant essence;

[0196] (7) Inspection and packaging:

[0197] Product inspection: sensory testing is conducted to check the texture, absorption and comfort of the serum to ensure that it is smooth, non-greasy and easily absorbed;

[0198] Cooling: Allow the essence to cool naturally to room temperature (25°C);

[0199] Packaging: Dispense the essence into glass bottles or aerosol cans, seal the packaging and ensure the hygienic conditions of the outer packaging;

[0200] Storage: Store in a cool, dry place away from direct sunlight.

[0201] Example 6: Soothing and repairing night cream

[0202] The anti-inflammatory aging composition comprises: 1g of sea cucumber oligopeptide, 1g of cysteine ​​(with a purity greater than 99%) and 0.8g of beta-cyclodextrin (with a purity greater than 99%), wherein the sea cucumber oligopeptide and cysteine ​​are cross-linked by beta-cyclodextrin to form the anti-inflammatory aging composition.

[0203] The preparation method of the anti-inflammatory aging composition is the same as that of Example 1, wherein the concentration of sea cucumber oligopeptide is 1 mg / mL, the concentration of cysteine ​​is 1 mg / mL, and the concentration of β-cyclodextrin is 0.8 mg / mL.

[0204] A soothing and repairing night cream comprises the following components in percentage by mass: 0.05% of the above-mentioned anti-inflammatory aging composition (for promoting collagen synthesis, enhancing skin elasticity, and resisting oxidation, slowing down the skin aging process, and reducing free radical damage), 5% glycerin (as a main moisturizer, with excellent hydration, helping to maintain skin moisture balance, and enhancing skin barrier function), 3% shea butter (shea butter is rich in fatty acids and vitamins, has a deep moisturizing effect, can effectively repair dry and cracked skin, and relieve skin tightness), 1% squalene (has a moisturizing effect, and helps to enhance skin barrier function, improve skin gloss and softness) and citric acid (as a pH regulator, and its amount is determined according to the target pH value), and the rest is deionized water.

[0205] The preparation method of the soothing and repairing night cream comprises the following steps:

[0206] (1) Raw material preparation: weigh each component according to the amount of each component in the above-mentioned soothing and repairing night cream and set aside;

[0207] (2) Preparation of aqueous phase: Add the anti-inflammatory aging composition to deionized water and stir at 200 rpm for 10 min using a stirrer to ensure that the components are evenly dispersed. It should be noted that the aqueous phase preparation process should be maintained at 25°C ± 2°C to avoid excessively high temperatures affecting the stability and activity of the components.

[0208] (3) Adding the oil phase: adding squalene and shea butter to the water phase and stirring at 300 rpm for 10 min to ensure that the oil phase and the water phase are fully blended to form a stable ointment;

[0209] (4) Ultrasonic treatment: The mixture is granulated using ultrasonic treatment technology to ensure that the particle size is between 50-500 nm to enhance permeability and bioavailability, wherein the ultrasonic power is set to 100 W and the treatment time is 10 min. It should be noted that during the ultrasonic treatment process, the temperature of the mixture is controlled at 25°C ± 2°C to avoid degradation of the components due to excessively high temperature.

[0210] (5) Adding glycerol: After the ultrasonic treatment, add glycerol and stir at 200 rpm for 10 min to ensure that the glycerol is completely dissolved and fully mixed with other ingredients;

[0211] (6) pH adjustment: Use citric acid to adjust the pH of the product to 5.5 to obtain a soothing and repairing night cream;

[0212] (7) Inspection and packaging:

[0213] Product inspection: sensory testing is conducted to check the texture, absorption and comfort of the night cream to ensure that it is smooth, non-greasy and easily absorbed;

[0214] Cooling: Allow the night cream to cool naturally to room temperature (25°C);

[0215] Packaging: Pack the night cream into glass bottles or other suitable containers, seal the packaging and ensure the hygienic conditions of the containers;

[0216] Storage: Store in a cool, dry place away from direct sunlight.

[0217] Example 7: Whitening Essence

[0218] The anti-inflammatory aging composition comprises: 0.8g sea cucumber oligopeptide, 1.5g cysteine ​​(purity greater than 99%) and 0.8g beta-cyclodextrin (purity greater than 99%), wherein the sea cucumber oligopeptide and cysteine ​​are cross-linked by beta-cyclodextrin to form the anti-inflammatory aging composition.

[0219] The preparation method of the anti-inflammatory aging composition is the same as that of Example 1, wherein the concentration of sea cucumber oligopeptide is 0.8 mg / mL, the concentration of cysteine ​​is 1.5 mg / mL, and the concentration of β-cyclodextrin is 0.8 mg / mL.

[0220] A whitening essence comprises the following components in percentage by mass: 0.8% of the above-mentioned anti-inflammatory aging composition (for promoting collagen synthesis, enhancing skin elasticity, and anti-oxidation, slowing down the skin aging process, and reducing free radical damage), 0.8% arbutin (arbutin is a natural whitening ingredient that can effectively inhibit the formation of melanin and reduce uneven skin color), 3% butylene glycol (as a moisturizer, which can increase skin hydration while improving skin transparency and gloss), 2% aloe extract (has soothing and anti-inflammatory effects, can calm the skin, and reduce discomfort caused by external stimuli) and citric acid (as a pH regulator, the amount of which is determined according to the target pH value), and the rest is deionized water.

[0221] The preparation method of the above whitening essence comprises the following steps:

[0222] (1) Raw material preparation: weigh each component according to the amount of each component in the above whitening essence and set aside;

[0223] (2) Preparation of aqueous phase: Add the anti-inflammatory aging composition to deionized water and stir with a stirrer at 200 rpm for 10 min until the ingredients are evenly dispersed; it should be noted that the aqueous phase preparation process should be maintained at 25°C ± 2°C to avoid high temperature damage to the active ingredients;

[0224] (3) Adding arbutin and aloe extract: Add arbutin and aloe extract to the aqueous phase and stir at 250 rpm for 5 min to ensure that the ingredients are evenly dispersed;

[0225] (4) Adding butanediol: Add butanediol and stir at 200 rpm for 10 min to ensure complete fusion;

[0226] (5) Ultrasonic treatment: The mixture is granulated using ultrasonic treatment technology to ensure that the particle size is between 30-300 nm to enhance permeability and bioavailability, wherein the ultrasonic power is set to 100 W and the treatment time is 10 min. It should be noted that during the ultrasonic treatment process, the temperature of the mixture is controlled at 25°C ± 2°C to avoid degradation of the components due to excessively high temperature.

[0227] (6) pH adjustment and filtration: Use citric acid to adjust the pH of the product to 4.5-5.5, and filter through a microporous membrane to remove undissolved particles or impurities to ensure the purity of the product and obtain a whitening essence;

[0228] (7) Inspection and packaging:

[0229] Product inspection: sensory testing to check the serum’s texture, absorption and comfort to ensure it is refreshing, non-greasy and easily absorbed;

[0230] Packaging: Dispense the essence into glass bottles or other suitable containers, seal the packaging and ensure the hygienic conditions of the containers;

[0231] Storage: Store in a cool, dry place away from direct sunlight.

[0232] Example 8: Deep Moisturizing Essence

[0233] The anti-inflammatory aging composition comprises: 0.5g sea cucumber oligopeptide, 0.5g cysteine ​​(purity greater than 99%) and 0.6g beta-cyclodextrin (purity greater than 99%), wherein the sea cucumber oligopeptide and cysteine ​​are cross-linked by beta-cyclodextrin to form the anti-inflammatory aging composition.

[0234] The preparation method of the anti-inflammatory aging composition is the same as that of Example 1, wherein the concentration of sea cucumber oligopeptide is 0.5 mg / mL, the concentration of cysteine ​​is 0.5 mg / mL, and the concentration of β-cyclodextrin is 0.6 mg / mL.

[0235] A deep moisturizing essence comprises the following components in percentage by mass: 0.5% of the above-mentioned anti-inflammatory aging composition (for promoting collagen synthesis, enhancing skin elasticity, and anti-oxidation, slowing down the skin aging process, and reducing free radical damage), 2% hyaluronic acid (as a powerful moisturizing ingredient, which can deeply replenish water, enhance skin elasticity and luster, and keep the skin hydrated), 1% vitamin C (has a strong antioxidant effect, can brighten skin tone, improve dullness, reduce melanin deposition, and promote collagen synthesis), 0.5% polyglycerol-6 (as a high-efficiency moisturizing ingredient, which can enhance skin barrier function and maintain long-term moisturizing effect) and citric acid (as a pH adjuster, the amount of which is determined according to the target pH value), and the rest is deionized water.

[0236] The preparation method of the above-mentioned deep moisturizing essence comprises the following steps:

[0237] (1) Raw material preparation: weigh each component according to the amount of each component in the above-mentioned deep moisturizing essence and set aside;

[0238] (2) Preparation of aqueous phase: Add the anti-inflammatory aging composition to deionized water and stir at 250 rpm for 10 min to ensure that the components are evenly dispersed. It should be noted that the aqueous phase preparation process should be maintained at 25°C ± 2°C to avoid excessively high temperatures affecting the stability and activity of the components.

[0239] (3) Adding hyaluronic acid and vitamin C: Add hyaluronic acid and vitamin C to the aqueous phase and stir until completely dissolved; the temperature of this step is controlled at 30°C ± 2°C to ensure that the ingredients are not damaged by heat and can be stably dissolved;

[0240] (4) Homogenization: The mixture is treated with a high shear homogenizer for 10 min to ensure that the particle size is between 100 and 300 nm and to form a stable emulsion;

[0241] (5) Adding polyglycerol-6: After homogenization, add butanediol and stir at 200 rpm for 10 min to ensure that polyglycerol-6 is fully blended with other ingredients;

[0242] (6) pH adjustment and filtration: Use citric acid to adjust the pH of the product to 4.5-5.5, and filter through a microporous membrane to remove undissolved particles or impurities to ensure the purity of the product and obtain a deep moisturizing essence;

[0243] (7) Inspection and packaging:

[0244] Product inspection: Conduct sensory testing to check the serum’s texture, whether it absorbs quickly and is non-greasy;

[0245] Packaging: Dispense the essence into glass bottles or other suitable containers, seal the packaging and ensure the hygienic conditions of the containers;

[0246] Storage: Store in a cool, dry place away from direct sunlight.

Claims

1. An anti-inflammatory aging composition, characterized in that: The invention comprises sea cucumber oligopeptide, cysteine ​​and beta-cyclodextrin. The sea cucumber oligopeptide and the cysteine ​​are cross-linked through the beta-cyclodextrin to form the anti-inflammatory aging composition.

2. The anti-inflammatory aging composition according to claim 1, characterized in that The mass ratio of the sea cucumber oligopeptide, the cysteine ​​and the β-cyclodextrin is (0.01-2):(0.01-2.5):(0.5-2).

3. The anti-inflammatory aging composition according to claim 1, characterized in that The mass ratio of the sea cucumber oligopeptide, the cysteine ​​and the β-cyclodextrin is (0.8-2):(0.5-1.5):(0.8-2).

4. The anti-inflammatory aging composition according to claim 1, characterized in that The mass ratio of the sea cucumber oligopeptide, the cysteine ​​and the beta-cyclodextrin is 1:1:

1.

5. The anti-inflammatory aging composition according to any one of claims 1 to 4, characterized in that: The molecular weight of the sea cucumber oligopeptide is 600-2000Da.

6. The method for preparing the anti-inflammatory aging composition according to any one of claims 1 to 5, characterized in that: The following steps are involved: dissolving the sea cucumber oligopeptide in water to obtain a sea cucumber oligopeptide solution; dissolving cysteine ​​in the sea cucumber oligopeptide solution to obtain an intermediate composition solution; dissolving β-cyclodextrin in water to obtain a β-cyclodextrin solution; The beta-cyclodextrin solution is slowly added to the intermediate composition solution, and the mixture is stirred to perform a cross-linking reaction to obtain an anti-inflammatory aging composition.

7. The method for preparing the anti-inflammatory aging composition according to claim 6, characterized in that: The reaction temperature of the cross-linking reaction is 30-50° C., and the reaction time is 4-8 hours.

8. Use of the anti-inflammatory aging composition according to any one of claims 1 to 5 in the preparation of skin care products.

9. The use according to claim 8, characterized in that: The skin care product includes any one of toner, lotion, face cream, eye cream and essence.

10. The use according to claim 8, characterized in that: In the anti-inflammatory aging composition, the concentration of the sea cucumber oligopeptide is 0.01-2 mg / mL, the concentration of cysteine ​​is 0.01-2.5 mg / mL, and the concentration of β-cyclodextrin is 0.5-2 mg / mL; in the skin care product, the added amount of the anti-inflammatory aging composition is 0.05% to 1% of the quality of the skin care product.

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