Skin care active substance and skin care product

Abstract

Provided are a skin care active substance and skin care product. The skin care active substance comprises, in part by weight: 3-90 parts of an anti-wrinkle / firming agent, 0.1-15 parts of a humectant, and 0.05-2.4 parts of an additional ingredient. The anti-wrinkle / firming agent comprises a first anti-wrinkle / firming agent and / or a second anti-wrinkle / firming agent. The first anti-wrinkle / firming agent comprises at least one of acetyl hexapeptide-8, tripeptide-1, acetyl octapeptide-3, and snake tripeptide, and the second anti-wrinkle / firming agent comprises at least one of palmitoyl tripeptide-5, palmitoyl tripeptide-1, palmitoyl hexapeptide-12, acetyl tetrapeptide-11, and hexapeptide-3. The humectant comprises at least one of ceramide NP, hydrolyzed sodium hyaluronate, and sodium hyaluronate. The additional component comprises at least one of 0.05-0.4 part of a moisturizing / firming agent and 0.05-2 parts of a soothing / antiallergic agent by weight. The moisturizing / firming agent comprises at least one of hydrolyzed lupin protein and collagen XVII, and the soothing / antiallergic agent comprises a paeonia root extract. The skin care active substance is designed to protect the skin barrier.

Description

Skin care actives, skin care products

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority based on Chinese patent application 202410115384.5 filed on January 25, 2024, and all of its contents are incorporated herein by reference. Technical Field

[0003] The present application relates to the technical field of skin care, and in particular to skin care active ingredients and skin care products. Background Art

[0004] As the body's physiological functions continue to decline, the skin becomes damaged under the influence of adverse factors such as pollution, stress, and ultraviolet rays, leading to skin aging.

[0005] With the continuous improvement of living standards, consumers are becoming more aware of anti-aging. Currently, most firming and anti-aging products focus on moisturizing and free radical scavenging, which are relatively common skincare methods. Currently, these products either use synthetic polymers to achieve a firming effect but fail to address skin aging, or use small-molecule peptide active ingredients that offer some anti-aging benefits but lack firming properties.

[0006] Summary of the Invention

[0007] The present application provides a skin care active ingredient and skin care product, which has both anti-aging and firming effects, effectively protects the skin barrier, and maintains skin health.

[0008] In order to solve the above technical problems, the first aspect of the present application provides a skin care active material, which comprises 3-90 parts by weight of an anti-wrinkle and firming agent, 0.1-15 parts by weight of a moisturizer, and 0.05-2.4 parts by weight of other ingredients; wherein the anti-wrinkle and firming agent comprises a first anti-wrinkle and firming agent and / or a second anti-wrinkle and firming agent, wherein the first anti-wrinkle and firming agent comprises at least one of acetyl hexapeptide-8, tripeptide-1, acetyl octapeptide-3, and snake venom peptide; and the second anti-wrinkle and firming agent comprises palmitoyl At least one of tripeptide-5, palmitoyl tripeptide-1, palmitoyl hexapeptide-12, acetyl tetrapeptide-11, and hexapeptide-3; the moisturizing agent includes at least one of ceramide NP, hydrolyzed sodium hyaluronate, and sodium hyaluronate; the other ingredients include 0.05-0.4 parts by weight of a moisturizing and firming agent, and 0.05-2 parts by weight of at least one of a soothing and anti-allergic agent; wherein the moisturizing and firming agent includes at least one of hydrolyzed lupine protein and XVII collagen, and the soothing and anti-allergic agent includes peony root extract.

[0009] In one embodiment, the skin care active ingredients include, by weight, at least one of: 3-10 parts by weight of acetyl hexapeptide-8, 3-10 parts by weight of tripeptide-1, 3-10 parts by weight of acetyl octapeptide-3, 3-10 parts by weight of snake venom peptide, 3-10 parts by weight of palmitoyl tripeptide-5, 3-10 parts by weight of palmitoyl tripeptide-1, 3-10 parts by weight of palmitoyl hexapeptide-12, 3-10 parts by weight of acetyl tetrapeptide-11, and 3-10 parts by weight of hexapeptide-3.

[0010] In one embodiment, the skin care active comprises, by weight, 0.5-5 parts by weight of ceramide NP, 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate; or

[0011] The skin care active ingredients include, by weight, 0.5-5 parts by weight of ceramide NP and 0.1-10 parts by weight of sodium hyaluronate; or

[0012] The skin care active ingredients include, by weight, 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate; or

[0013] The skin care active ingredient comprises, by weight, 0.5-5 parts by weight of ceramide NP.

[0014] In one embodiment, the skin care active ingredient comprises, by weight, 0.05-0.2 parts by weight of hydrolyzed lupin protein and / or 0.05-0.2 parts by weight of XVII collagen.

[0015] In one embodiment, the skin care active is composed of the following components in the following weight ratios: 3-10 parts by weight of acetyl hexapeptide-8, 3-10 parts by weight of palmitoyl tripeptide-5, 0.05-0.2 parts by weight of hydrolyzed lupin protein, 0.5-5 parts by weight of ceramide NP, 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate, and 0.05-2 parts by weight of peony root extract; or

[0016] The skin care active ingredient is composed of the following components in the following weight ratios: 0.5-5 parts by weight of ceramide NP, 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate, 3-10 parts by weight of acetyl hexapeptide-8, 3-10 parts by weight of palmitoyl tripeptide-5, and 0.05-0.2 parts by weight of hydrolyzed lupin protein; or,

[0017] The skin care active ingredient is composed of the following components in the following weight ratio: 0.5-5 parts by weight of ceramide NP, 3-10 parts by weight of acetyl hexapeptide-8, 3-10 parts by weight of palmitoyl tripeptide-5, and 0.05-0.2 parts by weight of hydrolyzed lupin protein; or,

[0018] The skin care active ingredient is composed of the following components in the following weight ratio: 0.5-5 parts by weight of ceramide NP, 0.1-10 parts by weight of sodium hyaluronate, 3-10 parts by weight of palmitoyl tripeptide-5, and 0.05-0.2 parts by weight of hydrolyzed lupin protein; or,

[0019] The skin care active ingredient is composed of the following components in the following weight ratios: 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate, 0.05-2 parts by weight of peony root extract, and 3-10 parts by weight of acetyl hexapeptide-8; or,

[0020] The skin care active ingredient is composed of the following components in the following weight ratios: 0.5-5 parts by weight of ceramide NP, 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate, 0.05-2 parts by weight of peony root extract, and 3-10 parts by weight of palmitoyl tripeptide-5.

[0021] In one embodiment, in the 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate, the weight ratio of the hydrolyzed sodium hyaluronate to the sodium hyaluronate is (2.5-4):(1.5-3).

[0022] In order to solve the above technical problems, the second aspect of the present application provides a skin care product, comprising any of the skin care active ingredients described above.

[0023] In one embodiment, the skin care product comprises 3.15-107.4 parts by weight of the skin care active ingredient and 7.5-14 parts by weight of the base material.

[0024] In one embodiment, the skin care product consists of the skin care active, the base and deionized water.

[0025] In one embodiment, the base material is composed of the following components in the following weight ratios: 3-5 parts by weight of glycerin, 2-3 parts by weight of polydimethylsiloxane, 1-3 parts by weight of squalane, 0.5-1 parts by weight of acrylic acid or acrylate or C10-30 alkyl acrylate crosspolymer, and 1-2 parts by weight of polysorbate 60.

[0026] Beneficial effects of the present application: Different from the prior art, the present application discloses a skin care active substance and a skin care product; the skin care active substance comprises 3-90 parts by weight of an anti-wrinkle and firming agent, 0.1-15 parts by weight of a moisturizer, and 0.05-2.4 parts by weight of other ingredients; wherein the anti-wrinkle and firming agent comprises a first anti-wrinkle and firming agent and / or a second anti-wrinkle and firming agent, the first anti-wrinkle and firming agent comprises at least one of acetyl hexapeptide-8, tripeptide-1, acetyl octapeptide-3, and a snake venom peptide; the second anti-wrinkle and firming agent comprises At least one of palmitoyl tripeptide-5, palmitoyl tripeptide-1, palmitoyl hexapeptide-12, acetyl tetrapeptide-11, and hexapeptide-3 is included in the active ingredients. The moisturizer includes at least one of ceramide NP, hydrolyzed sodium hyaluronate, and sodium hyaluronate. Other ingredients include 0.05-0.4 parts by weight of a moisturizing and firming agent and 0.05-2 parts by weight of at least one of a soothing and anti-allergic agent. The moisturizing and firming agent includes at least one of hydrolyzed lupine protein and collagen XVII, and the soothing and anti-allergic agent includes peony root extract. By designing the skincare active to include an anti-wrinkle and firming agent, it achieves both anti-wrinkle and firming benefits. Combined with the moisturizer, moisturizing and firming agent, and / or soothing and anti-allergic agent, it effectively protects the skin barrier and maintains skin health.

[0027] The above description is only an overview of the technical solution of the present application. In order to more clearly understand the technical means of the present application, it can be implemented in accordance with the contents of the specification. In order to make the above and other purposes, features and advantages of the present application more obvious and easy to understand, the specific implementation methods of the present application are listed below. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without any creative work.

[0029] FIG1 is a comparison chart of cell proliferation activity test results;

[0030] Figure 2 is a comparison of cell scratches;

[0031] Figure 3 is a comparison chart of cell scratch test results;

[0032] Figure 4 is a comparison of mitochondrial membrane potential;

[0033] FIG5 is a comparison diagram of mitochondrial membrane potential test results;

[0034] Figure 6 is a comparison chart of qPCR test results;

[0035] Figure 7 is a comparison of the experimental results of AQP-3;

[0036] Figure 8 is a comparison of the experimental results of ZO-1;

[0037] Figure 9 is a comparison of experimental results of Claudin-1;

[0038] FIG10 is a comparison chart of the experimental results of Collagen I;

[0039] FIG11 is a comparison chart of the experimental results of Collagen IV;

[0040] FIG12 is a comparison diagram of transmission electron microscopy experimental results;

[0041] FIG13 is a comparison diagram of the experimental results of HE staining;

[0042] FIG14 is a comparison chart of the experimental results of Masson staining. DETAILED DESCRIPTION

[0043] In order to make the purpose, technical solution and effect of this application clearer and more specific, the following embodiments of the technical solution of this application will be described in detail with reference to the accompanying drawings. The following embodiments are only used to more clearly illustrate the technical solution of this application and are therefore only examples and are not intended to limit the scope of protection of this application.

[0044] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this application belongs; the terms used herein are only for the purpose of describing specific embodiments and are not intended to limit this application; the terms "including" and "having" and any variations thereof in the specification and claims of this application and the above-mentioned figure descriptions are intended to cover non-exclusive inclusions.

[0045] In the description of the embodiments of the present application, the technical terms "first" and "second" are used only to distinguish different objects and should not be understood as indicating or implying relative importance or implicitly indicating the quantity, specific order, or primary and secondary relationship of the indicated technical features. In the description of the embodiments of the present application, the term "multiple" refers to more than two (including two), similarly, "multiple groups" refers to more than two (including two), and "multiple pieces" refers to more than two (including two), unless otherwise clearly and specifically defined.

[0046] References herein to "embodiments" mean that a particular feature, structure, or characteristic described in connection with the embodiments may be included in at least one embodiment of the present application. The appearance of this phrase in various places in the specification does not necessarily refer to the same embodiment, nor does it constitute an independent or alternative embodiment that is mutually exclusive of other embodiments. It is understood, both explicitly and implicitly, by those skilled in the art that the embodiments described herein may be combined with other embodiments.

[0047] In the description of the embodiments of this application, the term "and / or" is simply a description of the association relationship between associated objects, indicating that three relationships can exist. For example, A and / or B can represent the following three situations: A exists alone, A and B exist simultaneously, and B exists alone. In addition, the character " / " in this document generally indicates that the associated objects are in an "or" relationship.

[0048] Amounts, ratios, and other numerical values ​​are presented herein in a range format. It should be understood that such range format is used for convenience and brevity and should be interpreted flexibly to include not only the values ​​explicitly specified as range limits, but also all individual values ​​or sub-ranges encompassed within the range, as if each value and sub-range were explicitly specified.

[0049] If not otherwise specified, all steps of the present application may be performed sequentially, randomly, or in parallel, preferably sequentially. For example, the method includes steps (a) and (b), indicating that the method may include steps (a) and (b) performed sequentially, or may include steps (b) and (a) performed sequentially, or may include steps (a) and (b) performed simultaneously in parallel. For example, the method may further include step (c), indicating that step (c) may be added to the method in any order, for example, the method may include steps (a), (b), and (c), or may include steps (a), (c), and (b), or may include steps (c), (a), and (b), etc.

[0050] Skin aging is the outward manifestation of human aging, resulting from a combination of endogenous and exogenous factors. Endogenous aging refers to the natural aging of the skin with age, manifested by skin whitening, fine wrinkles, decreased elasticity, and sagging. Exogenous aging, the primary cause of photoaging caused by sun exposure, manifests as wrinkles, sagging, roughness, yellowish or grayish-yellow skin discoloration, dilated capillaries, and the formation of pigmentation spots.

[0051] In recent years, anti-aging awareness has been growing among consumers. According to survey data, the anti-aging market now accounts for 58% of the overall skincare market. Over 40% of women aged 20 to 24 use anti-aging products, with those born in the 1990s and 2000s becoming the primary drivers of this market.

[0052] Currently, most firming and anti-aging products are based on moisturizing and free radical scavenging, making them relatively common skincare methods. These products either use synthetic polymers to achieve a firming effect but fail to address skin aging, or they use small-molecule peptide active ingredients that offer some anti-aging benefits but lack firming properties. With limited options for firming and anti-aging products, some consumers are turning to cosmetic procedures to achieve wrinkle-lifting effects. However, cosmetic procedures often employ a process of first destroying and then rebuilding, which can damage the skin in a short period of time and is expensive.

[0053] In view of this, the present application provides a skin care active ingredient and a skin care product, which have both anti-wrinkle and firming effects, effectively protect the skin barrier and maintain skin health.

[0054] The present invention provides a skin care active ingredient. The skin care active ingredient includes, by weight, 3-90 parts of an anti-wrinkle and firming agent, 0.1-15 parts of a moisturizer, and 0.05-2.4 parts of other ingredients. The anti-wrinkle and firming agent includes a first anti-wrinkle and firming agent and / or a second anti-wrinkle and firming agent, wherein the first anti-wrinkle and firming agent includes at least one of acetyl hexapeptide-8, tripeptide-1, acetyl octapeptide-3, and a snake venom peptide; the second anti-wrinkle and firming agent includes at least one of palmitoyl tripeptide-5, palmitoyl tripeptide-1, palmitoyl hexapeptide-12, acetyl tetrapeptide-11, and hexapeptide-3; and the moisturizer includes at least one of ceramide NP, hydrolyzed sodium hyaluronate, and sodium hyaluronate. Other ingredients include 0.05-0.4 parts by weight of a moisturizing and firming agent and 0.05-2 parts by weight of at least one of a soothing and anti-allergic agent; wherein the moisturizing and firming agent includes at least one of hydrolyzed lupin protein and XVII collagen, and the soothing and anti-allergic agent includes peony root extract.

[0055] The first anti-wrinkle and firming agent acts on muscles, inhibiting muscle contraction and thus reducing the formation of wrinkles. For example, acetyl hexapeptide-8 can inhibit muscle contraction and thus reduce the formation of expression lines, thus achieving an anti-wrinkle effect and anti-aging through anti-wrinkle.

[0056] The second anti-wrinkle and firming agent acts on collagen in the skin, promoting its synthesis and maintenance, thereby reducing skin sagging and wrinkle formation. For example, palmitoyl tripeptide-5 can act on collagen in the skin, promoting its synthesis and maintenance, reducing skin sagging and wrinkle formation, thereby achieving an anti-wrinkle and firming effect. Simultaneously, palmitoyl tripeptide-5 can stimulate the proliferation of epidermal cells, promote the repair of the stratum corneum, and improve the skin's ability to retain moisture. In other words, palmitoyl tripeptide-5 has both anti-wrinkle and firming effects and moisturizing and repairing properties.

[0057] Ceramide NP is the main raw material for the brick wall structure of the skin barrier. It also stimulates the synthesis of endogenous sebum components to provide moisturizing and repair for dry and aging skin.

[0058] The functions of hydrolyzed sodium hyaluronate and sodium hyaluronate are to achieve moisturizing effects by strengthening the skin barrier and locking in moisture.

[0059] Moisturizing and firming agents stimulate collagen synthesis within cells, improving skin elasticity and firmness. They also have moisturizing properties, increasing skin moisture levels. For example, hydrolyzed lupin protein primarily promotes collagen production and skin firmness. This stimulates collagen synthesis within cells, improving skin elasticity and firmness. It also has moisturizing properties, increasing skin moisture levels and improving dryness and roughness.

[0060] Soothing and anti-allergic agents achieve soothing and anti-allergic effects through mitochondrial autophagy. For example, peony root extract optimizes cellular energy efficiency through mitochondrial autophagy, achieving soothing and anti-allergic effects, thereby combating skin aging.

[0061] By designing the skin care active ingredients to include at least one of a first anti-wrinkle and firming agent, a second anti-wrinkle and firming agent, and a soothing and anti-allergic agent with anti-aging effects, an anti-aging effect is achieved; by designing the skin care active ingredients to include at least one of a first anti-wrinkle and firming agent, a second anti-wrinkle and firming agent, and a moisturizing and firming agent with firming effects, a firming effect is achieved; and by designing the skin care active ingredients to include at least one of ceramide NP, hydrolyzed sodium hyaluronate, and sodium hyaluronate, a moisturizing effect is achieved. It can be understood that by designing the skin care active ingredients in this way, the skin care active ingredients simultaneously have anti-aging, firming, and moisturizing effects, effectively protecting the skin barrier and maintaining skin health.

[0062] In addition, the weight proportions of the various components of the skin care actives are designed as above, so that the skin care actives can act on the skin to have anti-wrinkle, firming, and moisturizing effects, and the effects are long-lasting without burdening the skin. It is relatively safe and can achieve anti-wrinkle, firming, and moisturizing effects gently and efficiently, promoting the skin to reach a healthy state.

[0063] It's important to note that healthy skin refers to skin that is smooth, soft, hydrated, even, firm, elastic, and flawless, based on the normal functioning of its physiological functions. Health is the foundation of beauty, and only healthy skin can achieve beauty. By empowering the skin with "balance, resilience, and self-healing abilities," we can gently and effectively maintain healthy skin.

[0064] Balance: The skin's surface and underlying layers, the stratum corneum and basal layer, each tightly bound together, enable the skin to maintain moisture and prevent water loss. The skin's ability to absorb and retain water, as well as the skin cells' ability to absorb and utilize oxygen, influence the skin's water balance and moisturizing capacity, as well as its metabolism and health, including its ability to heal itself.

[0065] Self-healing ability: The skin's ability to repair itself from damage, trauma, cell aging, etc., as well as the skin cells' ability to absorb and metabolize nutrients, affect the skin's metabolism and repair ability.

[0066] Resistance: The skin's ability to resist external environmental factors, including sunlight, pollutants, ultraviolet rays, bacteria, etc., as well as the skin cells' ability to identify and resist external pathogens and viruses, affect the health and appearance of the skin.

[0067] The acetyl hexapeptide-8, tripeptide-1, acetyl octapeptide-3, and snake venom-like peptide in this application correspond to "self-healing power", palmitoyl tripeptide-5, palmitoyl tripeptide-1, palmitoyl hexapeptide-12, acetyl tetrapeptide-11, and hexapeptide-3 correspond to "self-healing power", hydrolyzed lupin protein and XVII collagen correspond to "self-healing power", ceramide NP corresponds to "resistance", sodium hyaluronate and hydrolyzed sodium hyaluronate correspond to "balance power", and peony root extract corresponds to "self-healing power". The present application designs skin care active ingredients including anti-wrinkle and firming agents and moisturizers, wherein the anti-wrinkle and firming agents include at least one of acetyl hexapeptide-8, tripeptide-1, acetyl octapeptide-3, snake venom peptide, palmitoyl tripeptide-5, palmitoyl tripeptide-1, palmitoyl hexapeptide-12, acetyl tetrapeptide-11, and hexapeptide-3; the moisturizer includes at least one of ceramide NP, hydrolyzed sodium hyaluronate, and sodium hyaluronate; the skin care active ingredients also include at least one of hydrolyzed lupine protein, XVII collagen, and peony root extract, so that the skin care active ingredients have at least "self-healing power" and even "balancing power, resistance, and self-healing power" at the same time, thereby promoting the skin to a healthy state.

[0068] From another perspective, based on the framework of "balance, resistance, and self-healing," a combination of active ingredients derived from both human and plant sources can gently and efficiently achieve moisturizing, soothing, and anti-wrinkle benefits, promoting healthy skin. The ceramide NP, hydrolyzed sodium hyaluronate, sodium hyaluronate, acetyl hexapeptide-8, tripeptide-1, acetyl octapeptide-3, snake venom-like peptide, palmitoyl tripeptide-5, palmitoyl tripeptide-1, palmitoyl hexapeptide-12, acetyl tetrapeptide-11, hexapeptide-3, and XVII collagen in this application are ingredients derived from human sources, while peony root extract and hydrolyzed lupin protein are ingredients derived from plants. The present application designs skin care active ingredients including anti-wrinkle and firming agents and moisturizers, wherein the anti-wrinkle and firming agents include at least one of acetyl hexapeptide-8, tripeptide-1, acetyl octapeptide-3, snake venom peptide, palmitoyl tripeptide-5, palmitoyl tripeptide-1, palmitoyl hexapeptide-12, acetyl tetrapeptide-11, and hexapeptide-3; the moisturizer includes at least one of ceramide NP, hydrolyzed sodium hyaluronate, and sodium hyaluronate; the skin care active ingredients also include at least one of hydrolyzed lupine protein, XVII collagen, and peony root extract, so that the skin care active ingredients have both human homologous ingredients and plant-derived ingredients, and achieve moisturizing, soothing, and anti-wrinkle effects efficiently and gently.

[0069] In one embodiment, the skin care active ingredients include, by weight, at least one of: 3-10 parts by weight of acetyl hexapeptide-8, 3-10 parts by weight of tripeptide-1, 3-10 parts by weight of acetyl octapeptide-3, 3-10 parts by weight of snake venom peptide, 3-10 parts by weight of palmitoyl tripeptide-5, 3-10 parts by weight of palmitoyl tripeptide-1, 3-10 parts by weight of palmitoyl hexapeptide-12, 3-10 parts by weight of acetyl tetrapeptide-11, and 3-10 parts by weight of hexapeptide-3. It can be understood that the anti-wrinkle and firming agent of the skin care active substance is composed of a first anti-wrinkle and firming agent. When the first anti-wrinkle and firming agent is acetyl hexapeptide-8, the skin care active substance includes 3-10 parts by weight of acetyl hexapeptide-8; the anti-wrinkle and firming agent of the skin care active substance is composed of a second anti-wrinkle and firming agent. When the second anti-wrinkle and firming agent is palmitoyl tripeptide-5, the skin care active substance includes 3-10 parts by weight of palmitoyl tripeptide-5; the anti-wrinkle and firming agent of the skin care active substance is composed of a first anti-wrinkle and firming agent and a second anti-wrinkle and firming agent. When the first anti-wrinkle and firming agent is acetyl hexapeptide-8 and the second anti-wrinkle and firming agent is palmitoyl tripeptide-5, the skin care active substance includes 3-10 parts by weight of acetyl hexapeptide-8 and 3-10 parts by weight of palmitoyl tripeptide-5.

[0070] In one embodiment, the skin care active comprises 0.5-5 parts by weight of ceramide NPs and 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate. It will be appreciated that when the moisturizing agent for the skin care active comprises ceramide NPs, hydrolyzed sodium hyaluronate, and sodium hyaluronate, the skin care active comprises 0.5-5 parts by weight of ceramide NPs and 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate.

[0071] Wherein, 0.1-10 weight parts of hydrolyzed sodium hyaluronate and sodium hyaluronate refer to the total weight parts of hydrolyzed sodium hyaluronate and sodium hyaluronate in the skin care active material is 0.1-10 weight parts. In 0.1-10 weight parts of hydrolyzed sodium hyaluronate and sodium hyaluronate, the weight ratio of hydrolyzed sodium hyaluronate to sodium hyaluronate is (2.5-4): (1.5-3). Exemplary, in 0.1-10 weight parts of hydrolyzed sodium hyaluronate and sodium hyaluronate, sodium hyaluronate includes sodium hyaluronate with a molecular weight of 220 WDa (ten thousand Daltons), sodium hyaluronate with a molecular weight of 138 WDa (ten thousand Daltons), and sodium hyaluronate with a molecular weight of 36 WDa (ten thousand Daltons), and hydrolyzed sodium hyaluronate includes hydrolyzed sodium hyaluronate with a molecular weight of 3.9 WDa (ten thousand Daltons) and hydrolyzed sodium hyaluronate with a molecular weight of 7-8 KDa (kilo Daltons). 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate have hyaluronic acid of various molecular weights, covering the skin surface to the deep layer of the skin, strengthening the skin barrier function, locking in moisture, and achieving a moisturizing effect. The 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate in the following content are the same and will not be repeated. Exemplarily, in 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate, the weight ratio of hydrolyzed sodium hyaluronate to sodium hyaluronate can be 3.5:2.5, 3:2.5, 3:2, 3.5:2, etc.

[0072] In one embodiment, the skin care active comprises 0.5-5 parts by weight of ceramide NPs and 0.1-10 parts by weight of sodium hyaluronate. It will be appreciated that when the moisturizing agent for the skin care active comprises ceramide NPs and sodium hyaluronate, the skin care active comprises 0.5-5 parts by weight of ceramide NPs and 0.1-10 parts by weight of sodium hyaluronate.

[0073] In one embodiment, the skin care active comprises 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate. It will be appreciated that when the moisturizing agent for the skin care active comprises hydrolyzed sodium hyaluronate and sodium hyaluronate, the skin care active comprises 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate.

[0074] In one embodiment, the skin care active comprises 0.5-5 parts by weight of ceramide NPs. It is understood that when the moisturizing agent of the skin care active comprises ceramide NPs, the skin care active comprises 0.5-5 parts by weight of ceramide NPs.

[0075] In one embodiment, the skin care active comprises 0.05-0.2 parts by weight of hydrolyzed lupin protein and / or 0.05-0.2 parts by weight of XVII collagen. It is understood that when the moisturizing and firming agent is composed of hydrolyzed lupin protein, the skin care active comprises 0.05-0.2 parts by weight of hydrolyzed lupin protein.

[0076] In one embodiment, when the soothing and anti-allergic agent is composed of a peony root extract, the skin care active comprises 0.05-2 parts by weight of the peony root extract.

[0077] In one specific embodiment, the skin care active is composed of the following components in the following weight ratio: 3-10 parts by weight of acetyl hexapeptide-8, 3-10 parts by weight of palmitoyl tripeptide-5, 0.05-0.2 parts by weight of hydrolyzed lupin protein, 0.5-5 parts by weight of ceramide NP, 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate, and 0.05-2 parts by weight of peony root extract.

[0078] In one specific embodiment, the skin care active is composed of the following components in the following weight ratio: 0.5-5 parts by weight of ceramide NP, 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate, 3-10 parts by weight of acetyl hexapeptide-8, 3-10 parts by weight of palmitoyl tripeptide-5, and 0.05-0.2 parts by weight of hydrolyzed lupin protein.

[0079] In one embodiment, the skin care active ingredient is composed of the following components in the following weight ratio: 0.5-5 parts by weight of ceramide NP, 3-10 parts by weight of acetyl hexapeptide-8, 3-10 parts by weight of palmitoyl tripeptide-5, and 0.05-0.2 parts by weight of hydrolyzed lupin protein.

[0080] In one embodiment, the skin care active ingredient is composed of the following components in the following weight ratio: 0.5-5 parts by weight of ceramide NP, 0.1-10 parts by weight of sodium hyaluronate, 3-10 parts by weight of palmitoyl tripeptide-5, and 0.05-0.2 parts by weight of hydrolyzed lupin protein.

[0081] In one embodiment, the skin care active is composed of the following components in the following weight ratio: 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate, 0.05-2 parts by weight of peony root extract, and 3-10 parts by weight of acetyl hexapeptide-8.

[0082] In one specific embodiment, the skin care active is composed of the following components in the following weight ratio: 0.5-5 parts by weight of ceramide NP, 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate, 0.05-2 parts by weight of peony root extract, and 3-10 parts by weight of palmitoyl tripeptide-5.

[0083] It should be noted that when the skin care active includes at least one of acetyl hexapeptide-8, tripeptide-1, acetyl octapeptide-3, and snake venom-like peptide, the weight proportions of acetyl hexapeptide-8, tripeptide-1, acetyl octapeptide-3, and snake venom-like peptide in the skin care active can be 3 parts by weight, 4 parts by weight, 5 parts by weight, 6 parts by weight, 7 parts by weight, 8 parts by weight, 10 parts by weight, etc., respectively, or a range consisting of any two of the above values, for example, 3-5 parts by weight, 4-8 parts by weight, etc.

[0084] When the skin care active includes at least one of palmitoyl tripeptide-5, palmitoyl tripeptide-1, palmitoyl hexapeptide-12, acetyl tetrapeptide-11, and hexapeptide-3, the weight amounts of palmitoyl tripeptide-5, palmitoyl tripeptide-1, palmitoyl hexapeptide-12, acetyl tetrapeptide-11, and hexapeptide-3 in the skin care active can be 3 parts by weight, 4 parts by weight, 5 parts by weight, 6 parts by weight, 7 parts by weight, 8 parts by weight, 10 parts by weight, etc., respectively, or a range consisting of any two of the above values, for example, 3-5 parts by weight, 4-6 parts by weight, etc.

[0085] When the skin care active includes at least one of hydrolyzed lupine protein and collagen XVII, the weight amounts of hydrolyzed lupine protein and collagen XVII in the skin care active can be 0.05 parts by weight, 0.08 parts by weight, 0.11 parts by weight, 0.14 parts by weight, 0.17 parts by weight, 0.2 parts by weight, etc., respectively, or a range consisting of any two of the above values, for example, 0.05-0.14 parts by weight, 0.08-0.17 parts by weight, etc.

[0086] When the skin care active includes peony root extract, the weight portion of the peony root extract in the skin care active can be 0.05 weight part, 0.08 weight part, 0.11 weight part, 0.14 weight part, 0.17 weight part, 0.2 weight part, 0.6 weight part, 0.8 weight part, 1 weight part, 1.2 weight parts, 1.4 weight parts, 1.6 weight parts, 1.8 weight parts, 2 weight parts, etc., or a range consisting of any two of the above values, for example, 0.05-1 weight part, 0.14-1.4 weight parts, etc.

[0087] When the skin care active includes ceramide NPs, the weight amount of ceramide NPs in the skin care active can be 0.5 parts by weight, 1 part by weight, 1.5 parts by weight, 2 parts by weight, 2.5 parts by weight, 3 parts by weight, 3.5 parts by weight, 4 parts by weight, 4.5 parts by weight, 5 parts by weight, etc., or a range consisting of any two of the above values, such as 0.5-2 parts by weight, 2-4 parts by weight, etc.

[0088] When the skin care active includes hydrolyzed sodium hyaluronate and sodium hyaluronate, the total weight of hydrolyzed sodium hyaluronate and sodium hyaluronate in the skin care active can be 0.1 parts by weight, 0.5 parts by weight, 1 part by weight, 1.5 parts by weight, 2 parts by weight, 2.5 parts by weight, 3 parts by weight, 3.5 parts by weight, 4 parts by weight, 4.5 parts by weight, 5 parts by weight, 5.5 parts by weight, 6 parts by weight, 6.5 parts by weight, 7 parts by weight, 7.5 parts by weight, 8 parts by weight, 8.5 parts by weight, 9 parts by weight, 10 parts by weight, etc., or a range consisting of any two of the above values, for example, 0.1-5 parts by weight, 3-8 parts by weight, etc.

[0089] When the skin care active includes sodium hyaluronate, the weight amount of sodium hyaluronate in the skin care active can be 0.1 parts by weight, 0.5 parts by weight, 1 part by weight, 1.5 parts by weight, 2 parts by weight, 2.5 parts by weight, 3 parts by weight, 3.5 parts by weight, 4 parts by weight, 4.5 parts by weight, 5 parts by weight, 5.5 parts by weight, 6 parts by weight, 6.5 parts by weight, 7 parts by weight, 7.5 parts by weight, 8 parts by weight, 8.5 parts by weight, 9 parts by weight, 10 parts by weight, etc., or a range consisting of any two of the above values, for example, 0.1-4 parts by weight, 2-8 parts by weight, etc.

[0090] The embodiments of the present application also provide a skin care product, which includes the skin care active ingredients introduced in the above embodiments and has at least the same advantages as the above skin care active ingredients.

[0091] In one embodiment, the skin care product comprises, by weight, 3.15-107.4 parts of the skin care active ingredient and 7.5-14 parts of the base. The base typically includes a moisturizing ingredient. Optionally, the skin care product comprises, by weight, 3.15-37.2 parts of the skin care active ingredient. The composition and proportions of the skin care active ingredient are as described above and will not be further elaborated.

[0092] The combination of skin care actives and base materials forms a skin care product that can gently and effectively produce anti-wrinkle, firming and moisturizing effects, effectively protecting the skin barrier and maintaining skin health.

[0093] In one embodiment, the skin care product comprises a skin care active ingredient, a base, and deionized water. Optionally, the amount of deionized water to be added is calculated from the total weight of the skin care product being 100 parts by weight minus the sum of the weight of the skin care active ingredient and the weight of the base.

[0094] In one embodiment, the dosage form of the skin care product includes any one of a water-based solution, an emulsion, a cream, an essence lotion, a facial mask, and a spray. The skin care active ingredient in any of these dosage forms can achieve anti-wrinkle, firming, and moisturizing effects. The dosage form is selected based on specific needs; the base ingredients are designed based on the desired dosage form.

[0095] In one embodiment, the base is composed of the following components in the following weight ratios: 3-5 parts by weight of glycerin, 2-3 parts by weight of polydimethylsiloxane, 1-3 parts by weight of squalane, 0.5-1 parts by weight of acrylic acid, acrylic acid esters, or C10-30 alkyl acrylate crosspolymer, and 1-2 parts by weight of polysorbate 60. This base is suitable for use in serum emulsion formulations, where polysorbate 60 provides an emulsifying effect and acrylic acid esters provide a thickening effect.

[0096] In one embodiment, the skin care product comprises the following components in the following weight ratios: 3-10 parts by weight of acetyl hexapeptide-8, 3-10 parts by weight of palmitoyl tripeptide-5, 0.05-0.2 parts by weight of hydrolyzed lupin protein, 0.5-5 parts by weight of ceramide NP, 0.1-10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate, 0.05-2 parts by weight of peony root extract, 3-5 parts by weight of glycerin, 2-3 parts by weight of polydimethylsiloxane, 1-3 parts by weight of squalane, 0.5-1 parts by weight of acrylic acid, acrylates, or C10-30 alkyl acrylate crosspolymer, 1-2 parts by weight of polysorbate 60, and the remaining parts by weight of deionized water. The total weight of the skin care product is 100 parts.

[0097] The skin care actives provided herein have anti-wrinkle, firming, and moisturizing effects, and relevant experimental testing and research have been conducted at the cellular, biochemical, and skin membrane levels. The following skin care product, composed of the following components in the following weight ratio: 5 parts by weight of acetyl hexapeptide-8, 5 parts by weight of palmitoyl tripeptide-5, 0.1 parts by weight of hydrolyzed lupin protein, 1.5 parts by weight of ceramide NP, 1 part by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate (the weight ratio of hydrolyzed sodium hyaluronate to sodium hyaluronate is 3:2), 0.5 parts by weight of peony root extract, 5 parts by weight of glycerin, 2.3 parts by weight of polydimethylsiloxane, 2 parts by weight of squalane, 0.8 parts by weight of C10-30 alkyl acrylate crosspolymer, 1.6 parts by weight of polysorbate 60, and 75.2 parts by weight of deionized water, was used as an example. Experimental testing at the cellular, biochemical, and skin membrane levels was conducted.

[0098] 1. Cell proliferation activity assay.

[0099] Experimental steps:

[0100] (1) Cells were seeded into 96-well plates, and 100 μL of culture medium containing 8 × 103 cells was added to each well. The plates were incubated at 37°C for 24 h until the cells adhered to the plate and entered the logarithmic growth phase.

[0101] (2) After 24 hours, the culture medium in step (1) was discarded, and 100 μL of 0.05% concentration of the test solution was added to each well. Three parallel wells were set up and incubated for 24 hours.

[0102] (3) Aspirate the test solution added in step (2), add 90 μL of fresh culture medium and 10 μL of CCK-8 solution (Cell Counting Kit-8 reagent), and incubate for 1 to 4 hours.

[0103] (4) Measure the absorbance at 450 nm using an enzyme-labeled instrument.

[0104] Calculation method: Cell viability*(%)=[A(sample added)-A(blank) / [A(0 sample added)-A(blank)]×100;

[0105] Wherein, A(sample added) represents the absorbance of the wells containing cells, CCK-8 solution, and test solution;

[0106] A (blank) indicates the absorbance of wells with culture medium and CCK-8 solution but no cells;

[0107] A(0 loading) represents the absorbance of the wells containing cells and CCK-8 solution but no test solution;

[0108] Cell viability* indicates cell proliferation activity or cytotoxic activity.

[0109] It should be noted that when adding the culture medium containing CCK-8 to the cells in step (3), several blank wells without cells are added simultaneously to correct the values, which are referred to as A (blank). In step (2), several wells do not contain the test solution, but only the cell culture medium, which are referred to as A (0 addition).

[0110] Experimental results:

[0111] The experimental results are shown in Figure 1, which is a comparison chart of cell proliferation activity test results. As shown in Figure 1, the skin care products provided in the examples of this application can significantly promote cell proliferation and have significant pro-proliferation efficacy; specifically, the proliferation activity of HaCaT cells increased by 18.1% and that of HSF cells increased by 88.5%.

[0112] 2. Cell scratch test.

[0113] Experimental steps:

[0114] (1) HaCaT cells were seeded into 12-well plates and incubated at 37°C for 24 h. The cells adhered to the plate and entered the logarithmic growth phase.

[0115] (2) After 24 hours, the culture medium was discarded and 100 μL of 0.05% concentration of the test solution was added to each well. Three parallel wells were set up and incubated for 24 hours.

[0116] (3) Use a 100 μl pipette tip to make three parallel slits in each well, wash with PBS to remove the cells, add serum-free culture medium, take a photo under a microscope at 0 h, and continue to culture in the incubator for 24 h.

[0117] (4) After 24 hours, take photos under a microscope.

[0118] (5) Calculate the cell migration rate after 24 hours.

[0119] Experimental results:

[0120] The experimental results are shown in Figures 2 and 3. Figure 2 is a comparison diagram of cell scratches, and Figure 3 is a comparison diagram of cell scratch test results. As shown in Figures 2 and 3, the skin care products provided in the examples of the present application can significantly promote cell migration rate and have a significant repair effect; compared with the cell migration rate of the blank control group (pure culture medium, no test solution added) of 46.5%, the cell migration rate of the skin care product composition provided in the examples of the present application increased by 19.5%. It should be noted that in Figures 2 and 3, the positive control is the addition of EGF (growth factor, a substance that promotes migration).

[0121] 3. DPPH free radical scavenging experiment.

[0122] Experimental steps:

[0123] (1) Reagent preparation: Prepare a 150 μM DPPH (i.e., 150 μmol / L DPPH) solution in 80% ethanol.

[0124] (2) Sample addition: Add 20 μL of sample to each well, then add 180 μL of DPPH solution, and react at room temperature in the dark for 30 minutes.

[0125] (3) Detection: The absorbance at 517 nm was measured using an enzyme-labeled instrument, and the inhibition rate was calculated.

[0126] Calculation method: inhibition rate % = (1-A1 / A0) × 100%;

[0127] Wherein, A0 represents the absorbance value of the blank group at 517 nm; A1 represents the absorbance value of the sample group at 517 nm.

[0128] It should be noted that the sample is the skin care product of the present application. The blank group refers to the well in which no sample is added. In the DPPH free radical scavenging experiment, the skin care product provided in the embodiment of the present application, palmitoyl tripeptide-5, hydrolyzed lupine protein, ceramide NP, 5-fold hyaluronic acid (including sodium hyaluronate with a molecular weight of 220 WDa, sodium hyaluronate with a molecular weight of 138 WDa, sodium hyaluronate with a molecular weight of 36 WDa, hydrolyzed sodium hyaluronate with a molecular weight of 3.9 WDa, and hydrolyzed sodium hyaluronate with a molecular weight of 7-8 KDa), and peony root extract were used as samples in step (2) and the experiment was carried out in sequence. The concentration of Palmitoyl Tripeptide-5 is the same as the concentration of Palmitoyl Tripeptide-5 in the skin care active of the skin care product; the concentration of Hydrolyzed Lupin Protein is the same as the concentration of Hydrolyzed Lupin Protein in the skin care active of the skin care product; the concentration of Ceramide NP is the same as the concentration of Ceramide NP in the skin care active of the skin care product; the concentration of Quintuple Hyaluronic Acid is the same as the concentration and composition of Quintuple Hyaluronic Acid in the skin care active of the skin care product; and the concentration of Paeonia Lactiflora Root Extract is the same as the concentration of Paeonia Lactiflora Root Extract in the skin care active of the skin care product.

[0129] Experimental results:

[0130] The experimental results are shown in Table 1 below. The skin care active provided in the examples of the present application can significantly inhibit DPPH free radicals with a scavenging rate of 41.5%, and has significant antioxidant effect. The free radical scavenging rate of the skin care active provided in the examples of the present application is higher than that of each single component in the skin care active, reflecting the synergistic efficiency of the composition. It should be noted that the concentration of palmitoyl tripeptide-5 in the skin care active at a concentration of 10% is equivalent to 0.3%, the concentration of hydrolyzed lupin protein in the skin care active at a concentration of 10% is equivalent to 0.01%, the concentration of ceramide NP in the skin care active at a concentration of 10% is equivalent to 0.1%, the concentration of pentahedral hyaluronic acid in the skin care active at a concentration of 10% is equivalent to 0.1%, and the concentration of peony root extract in the skin care active at a concentration of 10% is equivalent to 0.1%.

[0131] Table 1 DPPH test results of skin care actives and active monomers of skin care actives

[0132] 4. Mitochondrial membrane potential detection experiment.

[0133] Experimental steps:

[0134] (1) Cell preparation: Cells were plated one day before the test to ensure that the cell confluence reached 50-70% at the time of the test.

[0135] (2) Sample addition: After 24 hours, the culture medium was discarded and 100 μL of 0.05% concentration of the test solution was added to each well. Three parallel wells were set up and incubated for 24 hours. The culture medium was prepared by mixing DMEM with fetal bovine serum in a ratio of 9:1.

[0136] (3) Aspirate the culture medium and wash the cells once with PBS or other appropriate solution if necessary according to the specific experiment.

[0137] (4) Drug induction: Remove the cell culture medium, dilute H2O2 to a concentration of 2 mmol / L with serum-free culture medium, and incubate in a cell culture incubator at 37°C in the dark for 4 hours.

[0138] (5) Aspirate the culture medium, wash the cells once with PBS, add 1 mL of JC-1 staining solution, and mix thoroughly. Incubate in a cell culture incubator at 37°C for 20 minutes. The JC-1 stock solution is 500X (that is, 500 times, the concentration of the staining solution is 1 times, that is, 1X, and the JC-1 staining solution is the 1X concentration after dilution).

[0139] (6) During the incubation period, prepare an appropriate amount of JC-1 staining buffer (1×) by adding 4 mL of distilled water per 1 mL of JC-1 staining buffer (5×) and place it in an ice bath.

[0140] (7) After incubation at 37°C, remove the supernatant and wash twice with JC-1 staining buffer (1×).

[0141] (8) Add 2 mL of cell culture medium containing serum and phenol red.

[0142] (9) Observe under a fluorescence microscope or laser confocal microscope.

[0143] Experimental results:

[0144] The experimental results are shown in Figures 4 and 5. Figure 4 is a comparison diagram of mitochondrial membrane potential, and Figure 5 is a comparison diagram of mitochondrial membrane potential test results. According to Figures 4 and 5, the skin care products provided in the examples of the present application can significantly reduce damage to mitochondria and have significant mitochondrial protective effects: compared with the fluorescence value of the blank control group of 0.63, the fluorescence value of the skin care composition provided in the examples of the present application is 0.54, and the protection rate of the composition increases by 14.2%. It should be noted that normal and healthy mitochondria are red fluorescence, and the model group is damaged by the addition of inducers to the mitochondria, and the red fluorescence is weakened. The sample group (skin care products of the present application) shows a protective effect by comparison with the model group.

[0145] 5. qPCR experiment.

[0146] Experimental steps:

[0147] (1) Cells were seeded into 24-well plates and incubated at 37°C for 24 h to allow the cells to adhere to the plate and enter the logarithmic growth phase.

[0148] (2) After 24 hours, the culture medium was discarded and 100 μL of 0.05% concentration of the test solution was added to each well. Three parallel wells were set up and incubated for 24 hours.

[0149] (3) Extract cellular RNA and reverse transcribe it into cDNA.

[0150] (4) Perform qPCR.

[0151] Experimental results:

[0152] The experimental results are shown in Figure 6, which is a comparison chart of the qPCR test results. As shown in Figure 6, the skin care products provided in the embodiment of the present application can significantly reduce the expression of MMP-1 (matrix metalloproteinase-1) and MMP-3 (matrix metalloproteinase-3), which are reduced by -36.5% and -17.3% respectively, and have a significant repair effect. It should be noted that the UVB model group damages cells through UVB, and the expression levels of MMP-1 and MMP-3 in the cells are increased, which will reduce the content of collagen; the sample group (the skin care products of the present application) is also irradiated with UVB. Compared with the model group, the expression levels of MMP-1 and MMP-3 genes are low, which weakens the decomposition of collagen, so that the skin care products of the present application can have a repair effect.

[0153] 6. Skin model.

[0154] Experimental Materials:

[0155] (1) Detection kit: T-Skin TM In vitro artificial full-thickness skin model test kit, batch number: 23TS060508.

[0156] (2) Culture conditions: 37°C, 5% CO2, relative humidity 95%.

[0157] (3) Culture medium: culture medium for full-thickness skin model (Lot: 23-MED-0505).

[0158] (4) Tissue fixative: 4% neutral paraformaldehyde solution.

[0159] (5)OCT frozen section embedding medium: Leica-14020108926.

[0160] (6) Anti-Collagen IV antibody: Abcam-ab6586.

[0161] (7) Recombinant Anti-Collagen I antibody: Abcam-ab138492.

[0162] (8) Recombinant Anti-Claudin I antibody: Abcam-ab211737.

[0163] (9) Anti-Collagen III antibody: Abcam-ab7778.

[0164] (10) Anti-Aquaporin 3 antibody: Abcam-ab125219.

[0165] (11) Recombinant anti-ZO1 tight junction protein antibody: Abcam-ab221547.

[0166] Experimental steps:

[0167] (1) Skin model preparation. After receiving the model, first preheat the accompanying culture medium. Add the preheated culture medium to the accompanying blue deep-well plate in advance, 8.5 mL / well. Use tweezers to remove the model from the test box, remove the residual agarose gel culture medium on the model, and then immediately transfer it to the blue deep-well plate that has been pre-added with culture medium. Each skin model is just stuck in the hole, and the bottom membrane of each skin model is in contact with the liquid surface to ensure that there are no bubbles between the model and the culture medium. Place the well plate in the incubator overnight for use.

[0168] (2) Raw material processing.

[0169] (a) Prepare a certain volume of culture medium containing the active substance according to the concentration.

[0170] (b) Remove the 6-well plate from the incubator, remove the culture medium from the bottom of the model using a pipette, and add 8.5 mL / well of fresh culture medium containing the active ingredient prepared in step (a). After the medium is replaced, ensure that the base membrane of each skin model is in contact with the liquid surface. Return the plate to the incubator and incubate at 37°C, 5% CO2 for 48 hours.

[0171] (3) Collection of skin model samples.

[0172] After the model incubation is completed (i.e., after step (2)), the model is carefully removed from the well plate with tweezers, the skin tissue is punched out with a punch, and the tissue is cut with a sharp blade. Tissues used for immunofluorescence experiments are fixed in OCT frozen section embedding medium. During the fixation process, bubbles in the OCT glue must be removed, and then quickly placed in liquid nitrogen. After marking the groups, transfer to -80°C for storage. Tissues used for transmission electron microscopy experiments are fixed in electron microscopy fixative and fixed at room temperature for more than 2 hours. Tissues used for HE staining and Masson staining are fixed in 4% paraformaldehyde.

[0173] (4) Tissue immunofluorescence.

[0174] Experimental process:

[0175] Allow frozen sections to dry at room temperature for 15 minutes. Use an immunohistochemistry pen to circle the tissue to be stained and soak in PBS for 10 minutes to remove the OCT cryosection embedding medium. Block sections with PBS containing 10% serum at room temperature. Apply various concentrations of primary antibody to the sections and incubate overnight at 4°C. Rinse sections three times with PBS for 10 minutes each. Add various concentrations of secondary antibody and incubate at 37°C for 1-2 hours. Rinse sections three times with PBS for 10 minutes each. Finally, mount the sections and observe the results under a fluorescence microscope and photograph.

[0176] (5) Transmission electron microscopy.

[0177] Experimental process:

[0178] (a) Washing: Rinse the sliced ​​tissue with 0.1 mol / L phosphate buffer 6 times, 30 minutes each time.

[0179] (b) Post-fixation: fixation with 1% osmium phosphate (prepared by dissolving 1 gram of osmium phosphate crystals in 0.1 mol / L phosphate buffer) for 1.5-2 hours.

[0180] (c) Washing: Rinse with 0.1 mol / L phosphate buffer three times for 10 minutes each. Steps (b) to (c) were performed in a refrigerator at 4°C.

[0181] (d) Gradient dehydration: 30% ethanol for 10 minutes; 50% ethanol for 10 minutes; 70% ethanol for 10 minutes; 90% ethanol for 10 minutes; 100% ethanol for 15 minutes; 100% ethanol:100% acetone (1:1) for 10 minutes; 100% acetone, twice, 15 minutes each time.

[0182] (e) Resin penetration:

[0183] 100% acetone: Epon812 resin = 1:1, oven at 38°C for 3-4 hours.

[0184] 100% acetone: Epon 812 resin = 1:2, oven at 38°C for 3-4 hours.

[0185] Pure Epon812 resin, oven at 38°C for 3-4 hours.

[0186] (f) Embedding: Embedding using embedding molds and Epon812 pure resin.

[0187] (g) Curing: Heating and polymerization curing in an electric heated forced air drying oven.

[0188] 38℃, 6 hours; 60℃, 6 hours; 80℃, 12 hours.

[0189] (h) Semi-thin section positioning:

[0190] The skin model was cut into semi-thin sections one micron thick using a Leica ultramicrotome. The areas suitable for electron microscopy were observed using an Olympus optical microscope, and suitable areas were located and selected for ultrathin sections using a single-sided blade.

[0191] (i) Ultrathin sections:

[0192] The tissues were sliced ​​using a Leica ultramicrotome with a thickness of 50-70 nm.

[0193] (j) Uranium-lead double staining:

[0194] Stain with 2% uranyl acetate (2 g uranyl acetate dissolved in 100 mL 70% ethanol) for 30 minutes at room temperature. Rinse the sections with about 300 mL of double-distilled water. After absorbing the water with filter paper, bake under a baking lamp for 15 minutes to completely dry the sections.

[0195] Stain with 3% lead citrate (add 0.04 g of lead citrate to 100 mL of freshly boiled double-distilled water, followed by 0.1 mL of 10 mol / L NaOH, and shake until the precipitate dissolves and the solution becomes clear). Stain for 15 minutes at room temperature. Rinse sections with approximately 300 mL of double-distilled water. After blotting with filter paper, bake under a heat lamp for 15 minutes to thoroughly dry the sections. The sections are then ready for electron microscopy.

[0196] (k) Transmission electron microscopy observation: observe the target structure, adjust the electron microscope until the image is clear, and collect the required images.

[0197] (6) HE staining of sections.

[0198] The tissue is removed from the fixative, trimmed, dehydrated, and transparentized. The transparentized tissue block is then embedded in melted paraffin and allowed to harden before sectioning. After sectioning, the sections are spread and baked, then stained with hematoxylin. Finally, the stained sections are dehydrated, transparentized, mounted, and imaged under a microscope.

[0199] (7) Masson staining of sections.

[0200] The paraffin sections were dewaxed and then stained with Masson's staining.

[0201] (a) Hematoxylin staining: Use Weigert's iron hematoxylin in the Masson staining kit to stain for 5 minutes, wash with tap water, differentiate with 1% hydrochloric acid alcohol for a few seconds, rinse with tap water, and rinse with running water for a few minutes to return to blue.

[0202] (b) Ponceau staining: Stain with the ponceau acid fuchsin solution in the Masson staining kit for 5-10 minutes, then rinse quickly with distilled water. Phosphomolybdic acid treatment: Treat with the phosphomolybdic acid aqueous solution in the Masson staining kit for approximately 3-5 minutes.

[0203] (c) Aniline blue staining: Without washing, directly counterstain with the aniline blue solution in the Masson staining kit for 5 minutes.

[0204] (d) Differentiation: treatment with 1% icy CH3COOH for 1 min.

[0205] (e) Dehydration and Mounting: Dehydrate and transparentize the sections in 95% alcohol I for 5 min, 95% alcohol II for 5 min, anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, pure xylene I for 5 min, and pure xylene II for 5 min. Remove the sections from the xylene and allow them to dry briefly, then mount the sections with neutral gum. "95% alcohol I" and "95% alcohol II" refer only to the first use of alcohol and should not be reused; "anhydrous ethanol I" and "anhydrous ethanol II" refer only to the first use of anhydrous ethanol and should not be reused; and "pure xylene I" and "pure xylene II" refer only to the first use of pure xylene and should not be reused.

[0206] (f) Microscopic examination, image acquisition and analysis.

[0207] Experimental results:

[0208] (1)AQP-3.

[0209] The experimental results for AQP-3 are shown in Figure 7 and Table 2. Figure 7 is a comparison chart of the experimental results for AQP-3. The negative control group consisted of culture medium only, the positive control group consisted of 200 μM Ve, and the sample group consisted of culture medium containing the skin care product of the present application. The sample groups referred to below have the same meaning and are not further described.

[0210] Table 2 AQP-3 mean fluorescence intensity (Mean±SD)

[0211] (2)ZO-1.

[0212] The experimental results of ZO-1 are shown in Figure 8 and Table 3. Figure 8 is a comparison chart of the experimental results of ZO-1.

[0213] Table 3 ZO-1 mean fluorescence intensity (Mean±SD)

[0214] (3) Claudin-1.

[0215] The experimental results of Claudin-1 are shown in Figure 9 and Table 4. Figure 9 is a comparison chart of the experimental results of Claudin-1.

[0216] Table 4 Claudin-1 mean fluorescence intensity (Mean±SD)

[0217] (4)Collagen I.

[0218] The experimental results of Collagen I are shown in Figure 10 and Table 5. Figure 10 is a comparison chart of the experimental results of Collagen I.

[0219] Table 5 Collagen I mean fluorescence intensity (Mean±SD)

[0220] (5)Collagen IV.

[0221] The experimental results of Collagen IV are shown in Figure 11 and Table 6. Figure 11 is a comparison chart of the experimental results of Collagen IV.

[0222] Table 6 Collagen IV mean fluorescence intensity (Mean±SD)

[0223] (6) Transmission electron microscopy.

[0224] The transmission electron microscope photograph is shown in FIG12 , which is a comparison diagram of the experimental results of the transmission electron microscope.

[0225] (7) HE staining.

[0226] The experimental results of HE staining are shown in Figure 13 and Table 7. Figure 13 is a comparison chart of the experimental results of HE staining.

[0227] Table 7 Epidermal thickness of the full skin model (Mean±SD)

[0228] It should be noted that “*” in Table 7 indicates that the difference is statistically significant compared with NC (p < 0.05).

[0229] (8)Masson staining.

[0230] The experimental results of Masson staining are shown in Figure 14. Figure 14 is a comparison chart of the experimental results of Masson staining.

[0231] It should be noted that in the experiment of this application, the skin model test was carried out under the full skin model T-SkinTM. According to the above skin model test results, it can be concluded that:

[0232] 1) Transmission electron microscopy revealed that after the skincare product was applied to T-SkinTM at a concentration of 0.6 mg / mL for 48 hours, the number of hemidesmosomes observed by transmission electron microscopy (TEM) was significantly increased in the sample group compared to the negative control group. These results indicate that the skincare product strengthens the dermal-epidermal junction (DEJ) and has a firming effect.

[0233] IF results showed that after 48 hours of exposure to the skincare product at a concentration of 0.6 mg / mL, AQP-3, ZO-1, Claudin I, Collagen I, and Collagen IV all showed fluorescence intensity. The relative fluorescence intensity of AQP-3 was 192.49%; that of ZO-1 was 152.38%; that of Claudin I was 173.11%; that of Collagen I was 241.18%; and that of Collagen IV was 280.22%. These results demonstrate that the skincare product has both repairing and anti-wrinkle effects.

[0234] 3) HE and Masson staining results showed that after 48 hours of exposure to T-SkinTM at a concentration of 0.6 mg / mL, the epidermal thickness of the sample increased significantly by 365.01 μm (p < 0.5) compared to the negative control. The number of collagen fibers in the dermis also increased, suggesting that the composition has a repairing effect.

[0235] Through testing at the cellular, biochemical, and skin model levels, including cell proliferation rate, cell migration rate, DPPH scavenging rate, mitochondrial activity, MMP-1, MMP-3, AQP-3, ZO-1, Claudin I, Collagen I, and Collagen IV, the skin care actives in the skin care products provided in the examples of this application were verified to effectively promote moisturizing, anti-wrinkle, and repair effects. The DPPH free radical scavenging rate of the skin care actives was higher than that of each individual component, demonstrating the synergistic and efficient nature of the skin care actives.

[0236] Experimental studies were conducted on the components and amounts of the skin care actives provided herein. The differences between the examples are as follows: the components and amounts of the skin care actives, as shown in Table 8, are identical in the base material used in each example, specifically: 4 parts by weight of glycerin, 2 parts by weight of polydimethylsiloxane, 1 part by weight of squalane, 1 part by weight of acrylates, 2 parts by weight of polysorbate 60, and the remaining parts by weight of deionized water, for a total of 100 parts by weight. The parts by weight of sodium hyaluronate and hydrolyzed sodium hyaluronate in each example in Table 8 are the total parts by weight, and the weight ratio of hydrolyzed sodium hyaluronate to sodium hyaluronate is 3:2.

[0237] Table 8 Test parameters of various embodiments

[0238] As can be seen from Examples 1 to 14 above, the skin care actives provided in the present application have firming, anti-wrinkle, and repairing effects. It should be noted that the specific testing methods for HSF cell proliferation activity and MMP-1 expression rate change in each example in Table 8 can be found in the relevant descriptions of the above examples and will not be repeated here.

[0239] The above description is only an implementation method of the present application and does not limit the patent scope of the present application. Any equivalent structure or equivalent process transformation made using the contents of the description and drawings of this application, or directly or indirectly applied in other related technical fields, are also included in the patent protection scope of the present application.

Claims

1. A skin care active ingredient, wherein, the skin care active ingredient comprises, by weight ratio, 3 - 90 parts by weight of an anti-wrinkle and firming agent, 0.1 - 15 parts by weight of a moisturizing agent, and 0.05 - 2.4 parts by weight of other ingredients; wherein, the anti-wrinkle and firming agent comprises a first anti-wrinkle and firming agent and / or a second anti-wrinkle and firming agent, and the first anti-wrinkle and firming agent comprises at least one of acetyl hexapeptide-8, tripeptide-1, acetyl octapeptide-3, and syn-ake; the second anti-wrinkle and firming agent comprises at least one of palmitoyl tripeptide-5, palmitoyl tripeptide-1, palmitoyl hexapeptide-12, acetyl tetrapeptide-11, and hexapeptide-3; the moisturizing agent comprises at least one of ceramide NP, sodium hyaluronate hydrolyzate, and sodium hyaluronate; the other ingredients comprise at least one of 0.05 - 0.4 parts by weight of a moisturizing and firming agent and 0.05 - 2 parts by weight of a soothing and anti-allergic agent; wherein, the moisturizing and firming agent comprises at least one of hydrolyzed lupetide and collagen XVII, and the soothing and anti-allergic agent comprises paeonia root extract.

2. The skin care active substance according to claim 1, wherein, The skin care active ingredient comprises, by weight ratio: at least one of 3 - 10 parts by weight of acetyl hexapeptide-8, 3 - 10 parts by weight of tripeptide-1, 3 - 10 parts by weight of acetyl octapeptide-3, 3 - 10 parts by weight of syn-ake, 3 - 10 parts by weight of palmitoyl tripeptide-5, 3 - 10 parts by weight of palmitoyl tripeptide-1, 3 - 10 parts by weight of palmitoyl hexapeptide-12, 3 - 10 parts by weight of acetyl tetrapeptide-11, and 3 - 10 parts by weight of hexapeptide-3.

3. The skin care active substance according to claim 1, wherein, The skin care active ingredient comprises, by weight ratio: 0.5 - 5 parts by weight of ceramide NP, 0.1 - 10 parts by weight of sodium hyaluronate hydrolyzate, and sodium hyaluronate; or, The skin care active ingredient comprises, by weight ratio: 0.5 - 5 parts by weight of ceramide NP, 0.1 - 10 parts by weight of sodium hyaluronate; or, The skin care active ingredient comprises, by weight ratio: 0.1 - 10 parts by weight of sodium hyaluronate hydrolyzate, and sodium hyaluronate; or, The skin care active ingredient comprises, by weight ratio: 0.5 - 5 parts by weight of ceramide NP.

4. The skin care active substance according to claim 1, wherein The skin care active ingredient comprises, by weight ratio: 0.05 - 0.2 parts by weight of hydrolyzed lupetide and / or 0.05 - 0.2 parts by weight of collagen XVII.

5. The skin care active substance according to claim 1, wherein, The skin care active ingredient consists of the following components by weight ratio: 3 - 10 parts by weight of acetyl hexapeptide-8, 3 - 10 parts by weight of palmitoyl tripeptide-5, 0.05 - 0.2 parts by weight of hydrolyzed lupetide, 0.5 - 5 parts by weight of ceramide NP, 0.1 - 10 parts by weight of sodium hyaluronate hydrolyzate, and sodium hyaluronate, 0.05 - 2 parts by weight of paeonia root extract; or, The skin care active ingredient consists of the following components by weight ratio: 0.5 - 5 parts by weight of ceramide NP, 0.1 - 10 parts by weight of sodium hyaluronate hydrolyzate, and sodium hyaluronate, 3 - 10 parts by weight of acetyl hexapeptide-8, 3 - 10 parts by weight of palmitoyl tripeptide-5, 0.05 - 0.2 parts by weight of hydrolyzed lupetide; or, The skin care active ingredient consists of the following components in the following weight ratios: 0.5 - 5 parts by weight of ceramide NP, 3 - 10 parts by weight of acetyl hexapeptide-8, 3 - 10 parts by weight of palmitoyl tripeptide-5, 0.05 - 0.2 parts by weight of hydrolyzed lupetide protein; or, The skin care active ingredient consists of the following components in the following weight ratios: 0.5 - 5 parts by weight of ceramide NP, 0.1 - 10 parts by weight of sodium hyaluronate, 3 - 10 parts by weight of palmitoyl tripeptide-5, 0.05 - 0.2 parts by weight of hydrolyzed lupetide protein; or, The skin care active ingredient consists of the following components in the following weight ratios: 0.1 - 10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate, 0.05 - 2 parts by weight of peony root extract, 3 - 10 parts by weight of acetyl hexapeptide-8; or, The skin care active ingredient consists of the following components in the following weight ratios: 0.5 - 5 parts by weight of ceramide NP, 0.1 - 10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate, 0.05 - 2 parts by weight of peony root extract, 3 - 10 parts by weight of palmitoyl tripeptide-5.

6. The skin care active substance according to claim 3 or 5, wherein In the 0.1 - 10 parts by weight of hydrolyzed sodium hyaluronate and sodium hyaluronate, the weight ratio of the hydrolyzed sodium hyaluronate to the sodium hyaluronate is (2.5 - 4):(1.5 - 3).

7. A skin care product, wherein, It includes the skin care active ingredient according to any one of claims 1 to 6.

8. The skin care product according to claim 7, wherein, The skin care product includes 3.15 - 107.4 parts by weight of the skin care active ingredient and 7.5 - 14 parts by weight of the base material according to the weight ratio.

9. The skin care product according to claim 8, wherein, The skin care product consists of the skin care active ingredient, the base material and deionized water.

10. The skin care product according to claim 7 or 8, wherein, The base material consists of the following components in the following weight ratios: 3 - 5 parts by weight of glycerol, 2 - 3 parts by weight of polydimethylsiloxane, 1 - 3 parts by weight of squalane, 0.5 - 1 part by weight of acrylic acid or acrylate or C10 - 30 alkanol acrylate cross-linked polymer, 1 - 2 parts by weight of polysorbate-60.

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