Cosmetic compositions and creams for firming the skin and mask liquids

Abstract

The application discloses a cosmetic composition and cream and mask liquid for skin tightening, which comprises: an active ingredient, which comprises at least one of the following components: icariin, ligustrazine, dihydromyricetin, an organic acid represented by formula (1); and one or more dispersion mediums for skin. The composition has good skin tightening activity, can significantly increase the collagen content in cells at a safe dose, can reduce the content of collagen degradation enzyme-matrix metalloproteinase-1 (MMP-1) to different degrees, and has a clear effect of increasing the skin tightening degree.

Description

Technical Field

[0001] This application belongs to the field of cosmetic technology, and particularly relates to a cosmetic composition, face cream, and face mask liquid for firming skin. Background Technology

[0002] Skin laxity is mainly characterized by loss of skin elasticity, loosening, partial collapse of skin structure, and the formation of wrinkles. The causes primarily include natural laxity and external factors. Natural laxity is mainly caused by damage to the extracellular matrix of dermal cells and decreased elastin expression. External factors contributing to laxity include ultraviolet radiation, smoking, and chemical damage.

[0003] Ultraviolet (UV) radiation is a major cause of skin laxity: ① UV radiation leads to structural changes in collagen, manifested as a decrease in its quantity or density; ② Fibroblasts and the extracellular matrix in the dermis adhere to each other, providing tension and elasticity to the skin, and the integrity of the collagen structure is fundamental to ensuring this function. These structural changes include: ① Increased collagen degradation. Matrix metalloproteinases (MMPs) are the main enzymes that degrade collagen and elastin in the dermis, and UV radiation accelerates their synthesis; ② Reduced collagen synthesis. UV radiation can inhibit the transforming growth factor-β (TGF-β) / Smad signaling pathway, reducing collagen synthesis and lowering the level of collagen in the body.

[0004] How to tighten the skin is a key research focus. Summary of the Invention

[0005] This application provides a cosmetic composition, cream, and mask liquid for skin firming, which has good skin firming activity, can significantly increase the collagen content in cells at a safe dose, and can reduce the content of collagen-degrading enzyme matrix metalloproteinase-1 (MMP-1) to varying degrees.

[0006] In a first aspect, embodiments of this application provide a composition for skin tightening, comprising:

[0007] The active ingredient includes at least one of the following components: icariin, tetramethylpyrazine, dihydromyricetin, and an organic acid having the structure of formula (1);

[0008]

[0009] Equation (1)

[0010] One or more dispersion media for use on the skin.

[0011] According to an embodiment of one aspect of this application, the active ingredients include icariin and dihydromyricetin in a molar ratio of 1:(0.1-1).

[0012] According to an embodiment of one aspect of this application, the active ingredients include tetramethylpyrazine and dihydromyricetin in a molar ratio of 1:(0.1-1).

[0013] According to one embodiment of this application, the molar concentrations of icariin, ligustrazine, dihydromyricetin, and organic acid in the composition are 0.01 to 5 μM, respectively.

[0014] According to one embodiment of this application, the dispersion medium includes any one or more media for ointments, creams, lotions, serums, patches, masks, tinctures.

[0015] According to one embodiment of this application, the dispersion medium is water.

[0016] Secondly, embodiments of this application provide a face cream for firming skin, comprising, by weight percentage: the active ingredient in the composition of the first aspect: 1%, stearic acid: 8%, glyceryl monostearate: 3%, olive oil: 3%, ammonium dimethyl taurate / VP copolymer: 6%, triethanolamine: 12%, glycerin: 18%, and the balance being water.

[0017] According to one embodiment of this application, the active ingredient is icariin.

[0018] Thirdly, embodiments of this application provide a facial mask liquid for firming skin, comprising, by mass percentage: the active ingredient in the composition of the first aspect: 0.01%, sodium carboxymethyl cellulose: 0.8%, glycerin: 16%, butylene glycol: 9%, aloe vera extract: 1.09%, phenoxyethanol: 1.2%, with the balance being deionized water.

[0019] According to one embodiment of this application, the active ingredients are icariin and dihydromyricetin, with a molar ratio of 1:(0.1-1).

[0020] The application embodiments have at least the following beneficial effects:

[0021] The natural products provided in this application's embodiments are: icariin, tetramethylpyrazine, dihydromyricetin, and organic acids. They have good skin-tightening activity and can significantly increase the collagen content in cells at safe doses. Further studies show that these compounds can increase the content of type I collagen, while all of them can reduce the content of collagen-degrading enzyme matrix metalloproteinase-1 (MMP-1) to varying degrees, thus having a clear effect on increasing skin firmness. They have a very broad prospect for application in the market of skin-tightening cosmetics. Attached Figure Description

[0022] To more clearly illustrate the technical solutions of the embodiments of this application, the drawings used in the embodiments of this application will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0023] Figure 1 The effects of icariin, tetramethylpyrazine, dihydromyricetin, organic acid (DES acid), and their combinations on collagen content in human embryonic skin fibroblasts (ESF) cells are illustrated in the figures (* represents p < 0.05 compared to the model group, ** represents p < 0.01 compared to the model group, *** represents p < 0.001 compared to the model group, **** represents p < 0.0001 compared to the model group, and the same applies below);

[0024] Figure 2 The cytotoxicity of icariin, tetramethylpyrazine, dihydromyricetin, and organic acid (DES acid) on ESF cells is illustrated in the embodiments of this application.

[0025] Figure 3 The effects of icariin, tetramethylpyrazine, dihydromyricetin, and organic acid (DES acid) on the expression level of type I collagen (COL-I) in human embryonic skin fibroblasts (ESF) cells are shown in the embodiments of this application.

[0026] Figure 4 The effects of icariin, tetramethylpyrazine, dihydromyricetin, and organic acid (DES acid) in embodiments of this application on the expression level of matrix metalloproteinase-1 (MMP-1) in human embryonic skin fibroblasts (ESF) cells are shown. Detailed Implementation

[0027] To make the inventive objectives, technical solutions, and beneficial technical effects of this application clearer, the following detailed description is provided in conjunction with specific embodiments. It should be understood that the embodiments described in this specification are merely illustrative and not intended to limit the scope of this application.

[0028] For simplicity, this paper only explicitly discloses some numerical ranges. However, any lower limit can be combined with any upper limit to form an unspecified range; and any lower limit can be combined with other lower limits to form an unspecified range, just as any upper limit can be combined with any other upper limit to form an unspecified range. Furthermore, although not explicitly stated, every point or individual value between the endpoints of a range is included within that range. Therefore, each point or individual value can serve as its own lower or upper limit and be combined with any other point or individual value, or with other lower or upper limits, to form an unspecified range.

[0029] In the description of this article, it should be noted that, unless otherwise stated, "above" and "below" include the number itself, and "several" in "one or several" means two or more.

[0030] The foregoing description of this invention is not intended to describe every disclosed embodiment or implementation. Instead, the following description provides more specific examples of exemplary embodiments. Throughout the application, guidance is provided through a series of embodiments that can be used in various combinations. The examples listed are merely representative and should not be construed as exhaustive.

[0031] The pursuit of beauty is universal, and firm skin is an important aspect of this pursuit. Research has found that skin firming can be achieved through the following methods: increasing collagen synthesis, promoting collagen fiber synthesis, and reducing the synthesis of matrix metalloproteinases (MMPs) in the skin, thereby slowing down collagen degradation. Collagen is the main substance supporting the structure of the epidermis and dermis; a decrease in its quantity or density is a major cause of skin structural collapse, reduced skin elasticity, and the formation of wrinkles. Collagen is an important structural component of the skin. Currently, it is known that the skin contains types I, III, IV, V, VII, XVII, and XVIII collagen. Types I and III collagen are found in the reticular fiber layer. Type IV collagen is a major component of the basement membrane. Type I collagen is mainly found in bone, tendons, ligaments, and skin tissue, while type III collagen is mainly found in blood vessels. Tissues containing type I collagen generally also contain type V collagen, while type II collagen is mainly found in cartilage tissue (which also contains type XI collagen) and the lens. Types IX, XII, and XIV collagen are also known as fibril-associated collagen (FAC) or fibril-associated collagen with interrupted triple-helices (FACIT). FACIT primarily coordinates interactions between fibril collagen molecules, between collagen and other cellular matrix components, and between cells. Type IX collagen is distributed on the surface of type II collagen microfibrils and forms covalent cross-linked structures with them. Types XII and XIV collagen are involved in the interaction with type I collagen molecules. Hyaluronic acid can minimize skin aging and wrinkles by absorbing and retaining moisture, resulting in a temporary plump and rounded appearance of the skin. Supplementing with collagen and hyaluronic acid can improve skin laxity and achieve skin firming. The main active compounds that can increase collagen synthesis and tighten the skin include: chemically synthesized peptides—palmitoyl tripeptide-5 and palmitoyl tetrapeptide-3; animal peptides—sheep placental peptides; vitamin C; α-hydroxy acids; and ethanol extracts of eucalyptus and sandalwood.

[0032] Collagen synthesis begins in the rough endoplasmic reticulum (RER). After synthesis on the RER, collagen α-chains undergo hydroxylation of proline and lysine residues via membrane-bound proteases—prolyl-4-hydroxylase and prolyl-3-hydroxylase. The C- and N-terminal propeptide sequences of the three α-chains form interchain cross-links via disulfide bonds, forming procollagen. Collagen peptidases remove the "loose" structures at both ends of procollagen, forming procollagen. Lysyl oxidase catalyzes the formation of covalent bonds between lysine and hydroxylysine, thereby polymerizing procollagen to form collagen fibers. Compounds that promote collagen fiber synthesis include retinol and marigold methanol extract.

[0033] Reducing the synthesis of matrix metalloproteinases (MMPs) helps slow down collagen degradation: Collagen degradation occurs through matrix metalloproteinases (MMPs). MMPs are a family of endopeptidases whose biological activity depends on zinc ions and has the ability to degrade the extracellular matrix (ECM). The main function of MMPs is to degrade collagen and elastin in the dermis. Their tissue inhibitors of metalloproteinases (TIMPs) specifically inhibit the activity of MMPs by covalently binding to their highly conserved zinc binding sites, jointly regulating matrix metabolism and maintaining the structure of the dermis. Increased MMP secretion or decreased TIMP secretion both accelerate collagen degradation, resulting in collagen loss in the skin, skin laxity, and decreased elasticity. Under physiological conditions, matrix metalloproteinase-1 (MMP-1) and MMP-8 break down the triple-helix structure of type I collagen into gelatin fragments of 1 / 4 and 3 / 4 length, respectively, which are then further degraded by MMP-2 and MMP-9. MMP-1, MMP-8, and MMP-13 are the main degrading enzymes of type I, II, III, and IV collagen. Type I collagen, cleaved into 1 / 4 and 3 / 4 fragments by these enzymes, is further cleaved by MMP-2 and MMP-9. Stromelysin degrades type IV collagen and fibronectin, while MMP-7 and MMP-12 degrade elastin. Tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) inhibits the activity of all MMPs, while Tissue inhibitor of matrix metalloproteinases-2 (TIMP-2) primarily inhibits MMP-2. Compounds that function by regulating the MMP / TIMP ratio include marine collagen synthetin; tea polyphenols, rhubarb extract, purslane extract, and eugenol, which have inhibitory effects on MMP-1.

[0034] Compositions for firming skin

[0035] In a first aspect, embodiments of this application provide a composition for skin tightening, comprising:

[0036] The active ingredient includes at least one of the following components: icariin, tetramethylpyrazine, dihydromyricetin, and an organic acid having the structure of formula (1);

[0037]

[0038] Equation (1)

[0039] One or more dispersion media for use on the skin.

[0040] Icariin is the main active ingredient of Epimedium, and is an 8-isopentenyl flavonoid glycoside. It can be extracted from the dried stems and leaves of Epimedium sagittatum, Epimedium pubescens, Epimedium wushanense, and Epimedium koreanum. Icariin has a certain soothing effect on sensitive skin, inhibits melanin synthesis and tyrosinase activity in melanoma cells, thus exhibiting a whitening effect. This application creatively discovers the efficacy of icariin in increasing collagen synthesis and firming the skin.

[0041] Ligustrazine is one of the effective components of the traditional Chinese medicine Ligusticum chuanxiong. This application creatively discovers that ligustrazine can increase collagen in skin cells, thereby achieving a skin-firming effect.

[0042] Dihydromyricetin is a natural component found in plant materials such as Ampelopsis japonica leaves, and it has certain anti-inflammatory and anti-photodamage effects. This application creatively reports that dihydromyricetin can increase skin collagen content and tighten the skin.

[0043] The organic acid shown in formula (1), (17R*,20R*,24Z)-21,26,29-trihydroxy-3,4-secocycloart-4(28),24(25)-dien-3-oic acid, is a natural product found in gardenia (hereinafter referred to as organic acid (DES acid)). This application creatively discovers that it has the effect of tightening the skin, and its CAS number is 2067656-40-8.

[0044] According to the embodiments of this application, it has been discovered for the first time that the above four compounds and their combinations can increase the collagen content in human embryonic skin fibroblasts (ESF) cells, exhibiting a clear skin-tightening effect. These compounds and their combinations have significant positive effects on novel applications in the preparation of skin-tightening cosmetics. Compared with extracts, the above compounds have clearly defined components and significantly greater activity.

[0045] According to the embodiments of this application, the icariin, tetramethylpyrazine, dihydromyricetin, and organic acids provided in this application have the following advantages: 1. The raw materials are readily available and all come from natural raw materials that can be obtained in large quantities; 2. They have strong skin-tightening activity; 3. They have high safety.

[0046] According to embodiments of this application, the dispersion medium may include water, commonly used cosmetic additives such as humectants and preservatives.

[0047] In some alternative embodiments, the active ingredients include icariin and dihydromyricetin in a molar ratio of 1:(0.1-1).

[0048] According to the embodiments of this application, controlling the molar ratio of icariin and dihydromyricetin within the above-mentioned range is beneficial to improving the skin-tightening effect.

[0049] In some alternative embodiments, the active ingredients include tetramethylpyrazine and dihydromyricetin in a molar ratio of 1:(0.1-1).

[0050] According to the embodiments of this application, controlling the molar ratio of tetramethylpyrazine and dihydromyricetin within the above-mentioned range is beneficial to improving the skin-tightening effect.

[0051] In some alternative embodiments, the molar concentrations of icariin, ligustrazine, dihydromyricetin, and organic acids in the composition are 0.01 to 5 μM, respectively.

[0052] According to embodiments of this application, using the collagen content in human embryonic skin fibroblasts (ESF) as a model for evaluation, it was shown that these compound monomers can significantly increase the collagen content in cells at a safe concentration of 5 μM, exhibiting clear skin-tightening activity. By controlling the dosage of the above-mentioned components within the aforementioned range, it is beneficial to protect the skin and improve the skin-tightening effect.

[0053] In some alternative embodiments, the dispersion medium includes any one or more media used in ointments, creams, lotions, serums, patches, masks, and tinctures.

[0054] In some alternative implementations, the dispersion medium is water.

[0055] Secondly, embodiments of this application provide a face cream for firming skin, comprising, by weight percentage: the active ingredient in the composition of the first aspect: 1%, stearic acid: 8%, glyceryl monostearate: 3%, olive oil: 3%, ammonium dimethyl taurate / VP copolymer: 6%, triethanolamine: 12%, glycerin: 18%, and the balance being water.

[0056] According to the embodiments of this application, the face cream composed of the above-mentioned components has a good skin-tightening effect and is skin-friendly and non-allergenic.

[0057] In some alternative embodiments, the active ingredient is icariin.

[0058] Thirdly, embodiments of this application provide a facial mask liquid for firming skin, comprising, by mass percentage: the active ingredient in the composition of the first aspect: 0.01%, sodium carboxymethyl cellulose: 0.8%, glycerin: 16%, butylene glycol: 9%, aloe vera extract: 1.09%, phenoxyethanol: 1.2%, with the balance being deionized water.

[0059] According to the embodiments of this application, the facial mask liquid composed of the above components has a good skin-tightening effect and is skin-friendly and non-allergenic.

[0060] In some optional embodiments, the active ingredients are icariin and dihydromyricetin in a molar ratio of 1:(0.1-1). According to embodiments of this application, controlling the molar ratio of icariin and dihydromyricetin within the above range is beneficial to improving the skin-tightening effect of the mask liquid.

[0061] This application discloses for the first time the use of icariin, tetramethylpyrazine, and dihydromyricetin organic acids in skin tightening. Therefore, they can be used alone or in combination with other active ingredients or excipients to make cosmetics or pharmaceuticals, as long as the pharmaceuticals can be used in the field of skin tightening and related fields.

[0062] Example

[0063] The following embodiments describe the disclosure of this application in more detail. These embodiments are merely illustrative, as various modifications and variations will be apparent to those skilled in the art within the scope of the disclosure of this application. Unless otherwise stated, all parts, percentages, and ratios reported in the following embodiments are based on weight, and all reagents used in the embodiments are commercially available or synthesized by conventional methods and can be used directly without further processing, and the instruments used in the embodiments are commercially available.

[0064] Example 1: Effect of the compound on collagen content in human embryonic skin fibroblasts (ESF)

[0065] Four monomeric compounds, namely icariin, tetramethylpyrazine, dihydromyricetin, and (17R*,20R*,24Z)-21,26,29-organic acids, and their combinations, were tested to determine their activity against collagen content in human embryonic skin fibroblasts (ESF). The specific test methods are as follows:

[0066] Human embryonic skin fibroblasts (ESF) derived from the Peking Union Medical College Cell Resource Center were seeded in 96-well plates at a density of 5 × 10⁶ cells / well. 4Cells were divided into three groups: a negative control, a positive control, and a screening drug group. Each group had three replicates. Cells were cultured in DMEM medium containing 1% penicillin, streptomycin, and 10% fetal bovine serum for 24 hours. Then, the medium was replaced with DMEM medium containing the drug and incubated at 37°C in a CO2 incubator for 24 hours. The drug was then added and incubated for another 24 hours. Two 96-well plates with the same treatment were prepared and used for the following assays:

[0067] (1) Detection of collagen content using Sirius red picric acid staining method

[0068] In the embodiments of this application, the collagen is secreted by fibroblasts, aggregates into collagen fibers outside the cells, and is then bound together into collagen fibers by a small amount of binders (proteoglycans and glycoproteins).

[0069] Dyeing principle: Sirius red is an anionic strong acid dye that readily reacts with the basic groups in collagen fibers, specifically staining them red. After staining, the absorbance value at a wavelength of 540 nm is measured using an ELISA reader; a higher OD value indicates more collagen fibers have been produced.

[0070] After the drug treatment period ended, the culture medium was discarded, and 100 μL of methanol was added to each well. The cells were fixed at -20°C overnight. After overnight fixation, the methanol was removed from the 96-well plate at room temperature, and 200 μL of PBS was added to each well for washing twice. The PBS wash buffer was removed, and 150 μL of 0.1% Sirius crimson picric acid staining solution was added to each well. The plate was shaken thoroughly at room temperature for 1 hour. The 0.1% Sirius crimson picric acid staining solution was removed, and 200 μL of 0.1% glacial acetic acid was added to each well for washing three times, 5 minutes each time. The plate was then air-dried for 3 hours. 200 μL of 0.1 mol / L sodium hydroxide was added to each well, and the plate was shaken thoroughly at room temperature for 1 hour. Using 0.1 M sodium hydroxide as a blank, the absorbance (OD) value at 540 nm was measured using a microplate reader.

[0071] (2) Total protein quantification

[0072] Detection of Cu using 2,2'-dioctoctanic acid (BCA) 1+ The content of Cu 2+ It is reduced to Cu by proteins in an alkaline environment. 1 + Two BCA molecules interact with one copper ion (Cu). 1+ The reaction product, a purple color, is formed through chelation. This water-soluble complex exhibits strong absorbance at 562 nm, which is linearly related to the increase in protein concentration. The absorbance (OD) value was measured at 450 nm using a microplate reader, and the sample concentration was calculated by plotting a standard curve.

[0073] After the drug treatment time is over, discard the culture medium, add 200 μL of PBS to each well and wash three times; add 50 μL of IPA lysis buffer to each well, place in a microwell shaker, set to 1000 rpm, 5℃, 30 min to fully lyse the cells, and use the cell lysate for protein quantification. Follow the instructions of the protein quantification kit for specific operating procedures.

[0074] The results are expressed as the relative amount of collagen / total protein, such as Figure 1 As shown, from Figure 1 The results showed that icariin, tetramethylpyrazine, dihydromyricetin, and organic acids at a concentration of 5 μM all significantly increased the relative expression of collagen in ESF cells. Compared with the positive control drugs 2.27 mM VC and 20 μM ginsenoside Re, their effects were comparable or better, indicating that these compounds have a clear skin-tightening effect.

[0075] Example 2: Study on the toxicity of the compound to human embryonic skin fibroblasts (ESF)

[0076] Following the same method as in Exercise 1, 5000 ESF cells / well were seeded into 96-well plates and cultured for 24 h. Different concentrations of the drug were added to each well, along with a negative control group containing only basal medium (DMSO) prepared with fresh medium (DMEM basal medium containing 1% penicillin and streptomycin, 10% fetal bovine serum). Each concentration was replicated in 5 wells. After culturing at 37°C in a 5% CO2 incubator for 24 h, 10 μL of MTT was added to bring the final MTT concentration to 0.5%. The cells were then incubated for 4 h. Finally, 100 μL of DMSO was added to each well, and the absorbance was measured at 570 nm using a microplate reader. Cell viability (%) was calculated using the following formula:

[0077] Cell viability (%) = (OD value of drug group - OD value of blank group) / (OD value of normal group - OD value of blank group) × 100%

[0078] The results are as follows Figure 2 As shown, by Figure 2 It can be seen that icariin, tetramethylpyrazine, dihydromyricetin, and DES acid have no obvious cytotoxicity at concentrations of 0.5-40 μM and do not affect the collagen test results.

[0079] Example 3: Effects of the compound on type I collagen (COL-I) and matrix metalloproteinase-1 (MMP-1) in human embryonic skin fibroblasts (ESF) cells.

[0080] (1) Experimental methods

[0081] Treat ESF cells as follows:

[0082] Human embryonic skin fibroblasts (ESF) were seeded in 24-well plates at a density of 1 × 10⁻⁶. 5 Cells were cultured in DMEM medium containing 1% penicillin and streptomycin and 10% fetal bovine serum for 24 h. The medium was then replaced with drug-containing DMEM medium and incubated at 37°C in a 5% CO2 incubator for 24 h. The culture medium was discarded, and residual cells were washed with PBS. 150 μL of RIPA lysis buffer was added for 30 min to lyse the cells (on ice). The resulting mixture was centrifuged at 1500 × g at 4°C for 10 min, and the supernatant lysis buffer was collected. The levels of human COL-I and MMP1 in the lysis buffer were determined according to the relevant kit instructions.

[0083] (2) Principle of the Human Type I Collagenase and MMP-1 Combined Immunosorbent Assay Kit

[0084] The kit employs a sandwich method. Specific anti-human COL-I monoclonal antibody is coated onto 96-well microplates. Human COL-I standards and samples are added to the wells, allowing the human COL-I protein in both the standards and samples to bind to the anti-human COL-I antibody immobilized on the microplate. Biotinylated anti-human COL-I monoclonal antibody is then added. After washing away unbound biotinylated antibody, HRP-labeled streptavidin is added, followed by thorough washing. TMB substrate is then added for color development. TMB is converted to blue under the catalysis of peroxidase, and then to yellow under acidic conditions. The color intensity is positively correlated with the amount of human COL-I protein in the sample. The absorbance (OD value) is measured at 450 nm using a microplate reader, and the sample concentration is calculated by plotting a standard midline.

[0085] The detection of MMP-1 was consistent with that of the collagenases mentioned above.

[0086] As attached Figure 3 As shown, icariin, tetramethylpyrazine, dihydromyricetin, and DES acid can all increase the expression of type I collagen in ESF cells to varying degrees at a concentration of 5 μM, with significant or highly significant effects compared to the control group, demonstrating that these compounds have a skin-tightening effect.

[0087] From the appendix Figure 4 It is known that icariin, tetramethylpyrazine, dihydromyricetin, and DES acid can all reduce the expression of matrix metalloproteinase-1 to varying degrees at a concentration of 5 μM, thereby slowing down the degradation of collagen and having a clear skin-tightening effect.

[0088] Example 4. Preparation of Epimedium-based Skin-Tightening Cream

[0089] This embodiment provides a skin-firming face cream comprising, by weight percentage: 1% icariin, 8% stearic acid, 3% glyceryl monostearate, 3% olive oil, 6% ammonium dimethyl taurate / VP copolymer, 12% triethanolamine, 18% glycerin, and purified water to 100%. The emulsification time was set at 20 minutes, resulting in a face cream with a uniform, smooth, and glossy appearance, good consistency, spreadability, and physical stability.

[0090] Example 5. Preparation of Skin-Tightening Mask of Composition 1 (Icariin + Dihydromyricetin)

[0091] This embodiment provides a facial mask liquid, comprising, by mass percentage: 0.8% sodium carboxymethyl cellulose, 16% glycerin, 9% butylene glycol, 1.09% aloe vera extract, 0.01% icariin and dihydromyricetin in a molar ratio of 1:1, 1.2% phenoxyethanol, and 71.9% deionized water. pH testing showed the mask to be weakly acidic, effectively tightening the skin. When applied, a non-woven fabric sheet mask is soaked in the liquid, with each sheet containing 20g of the skin-tightening mask liquid. Sensory testing showed the product was non-irritating to the skin and met the quality standards for facial masks.

[0092] This application evaluated the activity of four naturally derived, high-purity monomeric compounds isolated from human embryonic skin fibroblasts (ESF) at safe dosages, based on activity screening and toxicity assessment. The research process was rigorous and highly reliable. The activity of some of the compositions was also determined, demonstrating their effectiveness.

[0093] Using human embryonic dermal fibroblasts (ESF) cells as a model, the skin-tightening activities of icariin, tetramethylpyrazine, dihydromyricetin, (17R*,20R*,24Z)-21,26,29-trihydroxy-3,4-secocycloart-4(28),24(25)-dien-3-oic acid, and their combinations were evaluated. The results showed that these monomeric compounds significantly increased collagen content in ESF cells at a safe concentration of 5 μM, demonstrating significant skin-tightening activity, with effects comparable to or superior to the positive control agents vitamin C (2.27 mM) and ginsenoside Re (20 μM). Composition 1 (icariin: dihydromyricetin = 1:1, total 5μM) and Composition 2 (tetramethylpyrazine + dihydromyricetin = 1:1, total 5μM) can also significantly increase the collagen content in ESF cells and have significant skin-tightening activity.

[0094] The cytotoxicity of the compounds to human embryonic skin fibroblasts (ESF) was evaluated using the MTT assay. Icariin, tetramethylpyrazine, dihydromyricetin, and organic acids did not show significant cytotoxicity at concentrations ranging from 0.5 to 40 μM.

[0095] The preliminary mechanism by which the compounds increase collagen was evaluated using ELISA. It was found that the compounds exert their effects by inhibiting matrix metalloproteinase-1 (MMP-1), a collagen degradation-related protein, and increasing the activity of type I collagen in human embryonic skin fibroblasts (ESF). Icariin, tetramethylpyrazine, dihydromyricetin, and organic acids, at a concentration of 5 μM, significantly increased type I collagen content and inhibited MMP-1 expression, demonstrating a clear therapeutic effect.

[0096] It should also be noted that the exemplary embodiments mentioned in this application describe methods or systems based on a series of steps or apparatus. However, this application is not limited to the order of the above steps; that is, the steps can be performed in the order mentioned in the embodiments, or in a different order, or several steps can be performed simultaneously.

[0097] The above description is merely a specific embodiment of this application, but the scope of protection of this application is not limited thereto. Any person skilled in the art can easily conceive of various equivalent modifications or substitutions within the technical scope disclosed in this application, and these modifications or substitutions should all be covered within the scope of protection of this application. Therefore, the scope of protection of this application should be determined by the scope of the claims.

Claims

1. Use of a composition in the preparation of a skin-firming cosmetic, said composition comprising: Active ingredients, including (17) R *,20 R *,twenty four Z )-21,26,29-trihydroxy-3,4-secocycloart-4(28),24(25)-dien-3-oic acid, the (17) R *,20 R *,twenty four Z The molar concentration of 21,26,29-trihydroxy-3,4-secocycloart-4(28),24(25)-dien-3-oic acid in the composition is from 0.01 to 5 μM. One or more dispersion media for use on the skin.

2. The use according to claim 1, characterized in that, The dispersion medium includes any one or more media used in ointments, creams, lotions, serums, patches, masks, and tinctures.

3. The use according to claim 2, characterized in that, The dispersion medium is water.

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