Skin care composition comprising pinane diol and at least one soluble collagen and use thereof

Abstract

The invention provides a skin care composition. The skin care composition comprises the following components in a physiologically acceptable medium: (a) pinane glycol; and (b) at least one soluble collagen, the mass ratio of the component (a) to the component (b) being (6-20): 1. The pinanediol and the at least one soluble collagen are compounded to achieve a synergistic effect, and the expression quantity of the type I collagen in the skin can be remarkably improved, so that the synthesis of the type I collagen in the skin can be remarkably promoted under the condition of using a relatively small amount of pinanediol and the at least one soluble collagen, and the skin quality is improved. Meanwhile, the problems of irritation and the like possibly caused by too high active matter dosage are avoided, and the purposes of low cost, high efficacy and high safety are achieved.

Description

Technical Field

[0001] The present application relates to the technical field of cosmetics, and in particular to a skin care composition comprising pinanediol and at least one soluble collagen and a use thereof. Background Art

[0002] Collagen is an abundant protein that exists in many tissues such as skin, bones, tendons, cartilage and blood vessels. It performs various functions through the high-level structure formed by protein polymerization and plays an important role in the life activities of cells, tissues and organs. Among them, type I collagen accounts for more than 80% of the human body's collagen and plays a role in supporting the skin structure. In the skin, type I collagen is the main component of the dermis, constituting collagen fibers, providing elasticity and firmness to the skin. With age, the synthesis rate of type I collagen decreases, while the degradation rate increases, resulting in the breakage and reduction of collagen fibers, which causes aging phenomena such as sagging skin, wrinkles, and dryness. Therefore, it is necessary to restart collagen synthesis in the body to effectively resist skin aging (especially in the dermis). Summary of the invention

[0003] After extensive research, the inventors found that the combination of pinanediol and at least one soluble collagen has a synergistic effect in promoting the expression of type I collagen in the skin, thereby effectively resisting skin aging, especially dermal aging.

[0004] The specific technical solutions of this application are as follows:

[0005] 1. A skin care composition comprising the following components in a physiologically acceptable medium:

[0006] (a) pinanediol; and

[0007] (b) at least one soluble collagen,

[0008] The mass ratio of component (a) to component (b) is (6-20):1.

[0009] 2. The skin care composition according to item 1, wherein the mass ratio of component (a) to component (b) is (9-19):1.

[0010] 3. The skin care composition according to item 1 or 2, wherein the component (b) comprises type III collagen.

[0011] 4. Use of the skin care composition according to any one of items 1 to 3 for increasing the expression of type I collagen in the skin.

[0012] 5. Use of the skin care composition according to any one of items 1 to 3 for improving skin elasticity and firmness.

[0013] 6. Use of the skin care composition according to any one of items 1 to 3 in resisting and / or alleviating skin aging.

[0014] 7. Use of a combination of pinanediol and at least one soluble collagen for increasing the expression of type I collagen in the skin.

[0015] 8. Use of a combination of pinanediol and at least one soluble collagen for improving skin elasticity and firmness.

[0016] 9. Use of a combination of pinanediol and at least one soluble collagen in resisting and / or alleviating skin aging.

[0017] 10. The use according to any one of items 7 to 9, wherein the mass ratio of pinanediol to at least one soluble collagen is (6-20):1.

[0018] Effects of the Invention

[0019] The present application has a synergistic effect after compounding pinanediol with at least one soluble collagen, which can significantly increase the expression of type I collagen in the skin. Therefore, when using a relatively small amount of pinanediol and at least one soluble collagen, the synthesis of type I collagen in the skin can be significantly promoted, while avoiding the irritation and other problems that may be caused by excessive use of active ingredients, thereby achieving the goals of low cost, high efficacy and high safety. DETAILED DESCRIPTION

[0020] The present application is described in detail below in conjunction with the described embodiments. Although specific embodiments of the present application are shown, it should be understood that the present application can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided to enable a more thorough understanding of the present application and to enable the scope of the present application to be fully communicated to those skilled in the art.

[0021] It should be noted that certain words are used in the specification and claims to refer to specific components. Those skilled in the art should understand that technicians may use different nouns to refer to the same component. This specification and claims do not use the difference in nouns as a way to distinguish components, but use the functional differences of components as the criterion for distinction. As mentioned throughout the specification and claims, "including" or "comprising" are open-ended terms and should be interpreted as "including but not limited to". The subsequent description of the specification is a preferred embodiment of the present application, but the description is based on the general principles of the specification and is not intended to limit the scope of the present application. The scope of protection of the present application shall be determined by the attached claims.

[0022] The present application provides a skin care composition comprising the following components in a physiologically acceptable medium:

[0023] (a) pinanediol; and

[0024] (b) at least one soluble collagen,

[0025] The mass ratio of component (a) to component (b) is (6-20):1.

[0026] In some preferred embodiments, the mass ratio of component (a) to component (b) is (9-19):1.

[0027] For example, the mass ratio (m) of the component (a) to the component (b) 组分(a) :m 组分(b) ) can be 6:1, 6.5:1, 7:1, 7.5:1, 8:1, 8.5:1, 9:1, 9.5:1, 10:1, 10.5:1, 11:1, 11.5:1, 12:1, 12.5:1, 13:1, 13.5:1, 14:1, 14.5:1, 15:1, 15.5:1, 16:1, 16.5:1, 17:1, 17.5:1, 18:1, 18.5:1, 19:1, 19.5:1, 20:1, etc.

[0028] The amounts of component (a) and component (b) to be used according to the present application can vary within a wide range to determine suitable amounts, depending on the desired dermatological effect.

[0029] In some embodiments, the component (a) is 0.001-2% by mass in the skin care composition.

[0030] For example, in terms of mass percentage in the skin care composition, the component (a) can be 0.001%, 0.003%, 0.005%, 0.007%, 0.009%, 0.01%, 0.011%, 0.012%, 0.013%, 0.014%, 0.015%, 0.016%, 0.017%, 0.018%, 0.019%, 0.02%, 0.03%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.5%, 2.0%, etc.

[0031] In some embodiments, the component (b) accounts for 0.0005-2% by mass in the skin care composition.

[0032] For example, the component (b) may be 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.011%, 0.0 12%, 0.013%, 0.014%, 0.015%, 0.016%, 0.017%, 0.018%, 0.019%, 0.02%, 0.03%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.5%, 2.0%, etc.

[0033] The soluble collagen is soluble collagen, and in the present application refers to type III collagen.

[0034] In some embodiments, the component (b) comprises type III collagen.

[0035] In some embodiments, the component (b) comprises recombinant type III collagen.

[0036] In the present application, the molecular weight of the component (b) may be 10k-100kDa, for example, 20k-80kDa, 30k-70kDa, etc.

[0037] In the present application, there is no limitation on the method for determining the molecular weight of component (b). A person skilled in the art may determine the molecular weight using conventional methods in the art, such as gel permeation chromatography (GPC).

[0038] In some specific embodiments, the component (b) can be purchased or prepared by conventional methods in the art.

[0039] In some specific embodiments, the component (b) can be prepared by the method described in patent CN202210739807.1.

[0040] In some more specific embodiments, the component (b) is recombinant type III collagen prepared by the method described in patent CN202210739807.1.

[0041] In some more specific embodiments, the component (b) is a recombinant type III collagen having an amino acid sequence as shown in SEQ ID NO.1, prepared by the method described in patent CN202210739807.1.

[0042] In some more specific embodiments, the component (b) is a recombinant type III collagen having an amino acid sequence as shown in SEQ ID NO.2, prepared by the method described in patent CN202210739807.1.

[0043] In some more specific embodiments, the component (b) is a recombinant type III collagen having an amino acid sequence as shown in SEQ ID NO.3, which is prepared by the method described in patent CN202210739807.1.

[0044] As physiologically acceptable media, their composition, usage amount, etc. can be selected by those skilled in the art according to their knowledge and the type of application scenario required.

[0045] In the present application, the physiologically acceptable medium is such as solvents, UV filters, antioxidants, emulsifiers, fillers, colorants, preservatives, fragrances, skin care active agents such as moisturizers, anti-aging agents, whitening agents and mixtures thereof.

[0046] Examples of the solvent include water, oils and fats, hexylene glycol, butanediol, and 1,3-propylene glycol.

[0047] As UV filters, for example, benzophenone, 4-butylmethoxydibenzoylmethane, octocrylene, ethylhexyl methoxycinnamate, ethylhexyl salicylate, phenylbenzimidazole sulfonic acid or homosalate may be used.

[0048] As the antioxidant, for example, tocopherol acetate, BHT, BHA and the like are mentioned.

[0049] Examples of the emulsifier include anionic emulsifiers, cationic emulsifiers, amphoteric emulsifiers, and nonionic emulsifiers.

[0050] As fillers, for example, silicon dioxide, mica, kaolin, oxides of zinc and titanium of natural or synthetic origin; calcium carbonate, magnesium carbonate and magnesium hydrogen carbonate; zinc stearate, magnesium stearate or lithium stearate, zinc laurate, magnesium myristate and the like.

[0051] Examples of the coloring agent include iron oxide, carbon black, titanium dioxide, zinc oxide, cochineal, carotene, and chlorophyll.

[0052] Examples of the preservative include phenoxyethanol, benzyl alcohol, benzoic acid, sorbic acid, and imidazolidinyl urea.

[0053] Examples of the flavor include citronellol and tricyclodecenyl acetate.

[0054] Examples of the moisturizer include mannitol, sodium hyaluronate, lactic acid, glycerin, urea, and propylene glycol.

[0055] As anti-aging agents, for example, retinol, phytosterols, ceramide, recombinant type I collagen, recombinant type II collagen, recombinant type IV collagen and recombinant type XVII collagen.

[0056] As whitening agents, there are arbutin, phenethylresorcinol, ascorbic acid and its derivatives and the like.

[0057] The present application provides the use of the above-mentioned skin care composition in increasing the expression of type I collagen in the skin.

[0058] The components (a) and (b) in the skin care composition work synergistically to increase the expression of type I collagen in the skin.

[0059] The present application provides the use of the above-mentioned skin care composition in improving skin elasticity and firmness.

[0060] The present application provides use of the above-mentioned skin care combination in resisting and / or alleviating skin aging.

[0061] The skin care composition described in the present application can effectively resist skin aging because it can increase the expression of type I collagen in the skin, thereby improving skin elasticity and skin firmness and resisting and / or alleviating skin aging.

[0062] The present application provides use of the above-mentioned skin care composition in preparing cosmetics.

[0063] The present application provides the use of a combination of pinanediol and at least one soluble collagen in increasing the expression of type I collagen in the skin.

[0064] The present application provides the use of a combination of pinanediol and at least one soluble collagen in improving skin elasticity and firmness.

[0065] The present application provides use of a combination of pinanediol and at least one soluble collagen in resisting and / or alleviating skin aging.

[0066] The definitions of pinanediol and soluble collagen and the mass relationship thereof are as described above.

[0067] The composition described in the present application is applied in the field of beauty and skin care to achieve the technical effects of the present application for non-therapeutic purposes.

[0068] Example

[0069] The present application provides a general and / or specific description of the materials and test methods used in the experiments. In the following examples, if there is no other special description, % means wt%, i.e., weight percentage. The reagents or instruments used without indicating the manufacturer are all conventional reagent products that can be obtained commercially. Among them, the content of pinanediol is about 100%, which is purchased from Shanghai Xianding Biotechnology Co., Ltd., and the recombinant type III collagen (the content of soluble collagen is about 100%) is purchased from Huaxi Biotechnology (Tianjin) Co., Ltd. (trade name: Bloomcolla TM ColⅢ recombinant type Ⅲ collagen).

[0070] Experimental Example 1 Cytotoxicity Test

[0071] 1. Experimental Materials

[0072] Human fibroblasts (Yimo Biotechnology, IM-H032), cell culture medium: low-glucose DMEM culture medium (VivaCell) + 10% fetal bovine serum (Gibco), PBS (VivaCell), MTT (Sigma), DMSO (Sinopharm), trypsin (Gibco).

[0073] 2. Experimental Methods

[0074] The experiment set up a zero-adjustment group, a control group, a positive control group and a sample group.

[0075] Sample group: 8 concentration gradients of pinanediol and soluble collagen were set as shown in Tables 1 and 2 respectively (preparation method: weigh 0.1 g of pinanediol sample, dissolve in 1 mL of anhydrous ethanol, prepare a 10% stock solution, and dilute with PBS; weigh 0.1 g of soluble collagen sample, dissolve in 1 mL of PBS, prepare a 10% stock solution, and dilute with PBS);

[0076] Control group: cell culture medium;

[0077] Positive control group: cell culture medium containing 10% DMSO;

[0078] Zero adjustment group: cell culture medium.

[0079] Three replicate wells were set up under each concentration gradient, and the drug was administered when the plating rate of human fibroblasts in the 96-well plate reached 40% to 60%. 200 μL of cell culture medium containing 10% PBS was added to each well of the control group; 200 μL of cell culture medium containing 10% DMSO was added to each well of the positive control group; 200 μL of cell culture medium containing the corresponding concentration of sample was added to each well of the sample group; the zero adjustment group had no cell inoculation and only 200 μL of cell culture medium was added. After the administration was completed, the 96-well plate was placed in an incubator (37°C, 5% CO2) for culture. After the cells were incubated for 24 hours, the supernatant was discarded, MTT working solution (0.5 mg / mL) was added, and the cells were incubated at 37°C in the dark for 4 hours. After the incubation was completed, the supernatant was discarded, 100 μL DMSO was added to each well, and the OD value was read at 490nm. The cell survival rate was calculated using the following formula: cell survival rate (%) = average absorbance (OD value) of the test group / average absorbance (OD value) of the blank control group × 100%.

[0080] When the cell survival rate was ≥90%, there was no obvious cytotoxicity, and the concentration without cytotoxicity was selected for the next test.

[0081] The results are shown in Table 1-2.

[0082] Table 1 Soluble collagen cytotoxicity results

[0083]

[0084] Table 2 Pinanediol cytotoxicity results

[0085]

[0086] As can be seen from Table 1-2, pinanediol has no cytotoxicity at a concentration below 0.03% and soluble collagen has no cytotoxicity at a concentration below 2.3%.

[0087] Experimental Example 2 Type I collagen gene expression test

[0088] 1. Experimental Materials

[0089] Human fibroblasts (Yimo Biotechnology, IM-H032), cell culture medium: low-glucose DMEM culture medium (VivaCell) + 10% fetal bovine serum (Gibco), PBS (VivaCell), trypsin (Gibco), cell lysis solution (RNA-easyIsolation Reagent Novozyme).

[0090] 2. Experimental Equipment

[0091] CO2 incubator (Thermo, 160i), biological safety cabinet (Sujing Antai, BSC-1604ⅡA2), inverted fluorescence microscope (Keyence BZ-X810), microplate reader (Tecan, Spark), floor-standing half-body ultraviolet therapy device (Sigma, SS-03AB), Q-PCR instrument (Thermo Fisher, ABI7500).

[0092] 3. Experimental methods:

[0093] Fibroblasts were used as the research object, UVB stimulation was used to establish an in vitro cell model, and the relative expression of type Ⅰ collagen gene in the cells was detected by q-PCR detection technology to explore the collagen-promoting effect of pinanediol, soluble collagen and their combination.

[0094] The experiment set up blank control group, negative control group, positive control group and sample group.

[0095] Blank control group: cell culture medium;

[0096] Negative control group: cell culture medium;

[0097] Positive control group: cell culture medium containing 100 μg / mL vitamin C (VC) and 7 μg / mL vitamin E (VE);

[0098] Sample set:

[0099] Pinanediol: weigh 0.1 g sample and dissolve it in 1 mL DMSO to prepare a 10% stock solution;

[0100] Soluble collagen: weigh 0.1 g sample and dissolve it in 1 mL PBS to prepare a 10% stock solution;

[0101] The samples of Examples 1-6 and Comparative Examples 1-8 were prepared using cell culture medium according to Table 3.

[0102] According to the appropriate inoculation density (8×10 4 / well) to inoculate cells into 24-well plates and incubate them in an incubator (37°C, 5% CO2) overnight to complete cell inoculation. When the cell plating rate in the 24-well plate reaches 40% to 60%, irradiation is performed. The total dose received by the negative control group, positive control group and sample group is 9J / cm 2 At the same time, the blank control group was placed in the same environment (UVA radiation dose of 0 J / cm 2 ). After the irradiation is completed, the drug is administered. 1 mL of cell culture medium is added to each well of the blank control group and the negative control group; 1 mL of cell culture medium containing 100 μg / mL vitamin C (VC) and 7 μg / mL vitamin E (VE) is added to each well of the positive control group; 1 mL of the sample of Examples 1-8 and Comparative Examples 1-6 is added to each well of the sample group.

[0103] After 24 hours of incubation, 0.5 mL of cell lysis buffer was added to each well and placed at room temperature for 5 minutes to fully lyse the cells. The cells were transferred to a 1.5 mL RNase-free Eppendorf tube and RNA was extracted according to the operating procedures. cDNA was synthesized according to the instructions of the RNA reverse transcription kit. qRT-PCR was performed to detect the type I collagen gene.

[0104] Table 3

[0105] Pinanediol content Soluble collagen content The quality ratio of the two Example 1 0.0095％ 0.0005％ 19:1 Example 2 0.019％ 0.001％ 19:1 Example 3 0.009375％ 0.000625％ 15:1 Example 4 0.01875％ 0.00125％ 15:1 Example 5 0.009％ 0.001％ 9:1 Example 6 0.018％ 0.002％ 9:1 Comparative Example 1 0.01％ - - Comparative Example 2 - 0.01％ - Comparative Example 3 0.02％ - - Comparative Example 4 - 0.02％ - Comparative Example 5 0.008％ 0.002％ 4：1 Comparative Example 6 0.016％ 0.004％ 4：1 Comparative Example 7 0.005％ 0.005％ 1:1 Comparative Example 8 0.0192％ 0.0008％ 24:1

[0106] 4. Statistical Analysis of Results

[0107] β-actin was used as the internal reference for gene expression to calculate the relative RNA expression of the target gene.

[0108]

[0109] ΔC(t)=C(t) 目的基因 –C(t) β-actin

[0110] All experiments were repeated three times, and the obtained data were expressed as Mean±SD. T.TEST test was used, and the difference was considered statistically significant when P-value < 0.05. The results are shown in Table 4.

[0111] If the effect of the combination is greater than that of a single substance, and P < 0.05, the effects of the two components can be considered synergistic; if the effect of the combination is weaker than that of a single substance, and P < 0.05, the effects of the two components can be considered antagonistic; if P > 0.05, the effects of the two components can be considered additive.

[0112] Table 4

[0113]

[0114]

[0115] When using the t-test two-tailed test method for statistical analysis, the significance of the NC group compared with the BC group is indicated by #, p-value < 0.05 is indicated by #, p-value < 0.01 is indicated by ##, and p-value < 0.001 is indicated by ###. The significance of the sample group and PC group compared with the NC group is indicated by *, p-value < 0.05 is indicated by *, p-value < 0.01 is indicated by **, and p-value < 0.001 is indicated by ***. The significance of the sample combination group compared with the sample pinanediol monomer group is indicated by ▲ indicates that p-value<0.05 indicates that▲ , p-value<0.01 is indicated as ▲▲ , p-value<0.001 is indicated as ▲▲▲ Compared with the sample soluble collagen monomer group, the sample combination group was significantly △ indicates that p-value<0.05 indicates that △ , p-value<0.01 is indicated as △△ , p-value<0.001 is indicated as △△△ .

[0116] It can be seen from the above table that the pinanediol and soluble collagen in the composition described in the example work synergistically to significantly increase the expression level of type I collagen gene.

[0117] The above is only the preferred embodiment of the present application, and does not limit the present application in other forms. Any technician familiar with the profession may use the above disclosed technical content to change or modify it into an equivalent embodiment with equivalent changes. However, any simple modification, equivalent change and modification made to the above embodiment based on the technical essence of the present application without departing from the technical solution of the present application still falls within the protection scope of the technical solution of the present application.

Claims

1. A skin care composition comprising the following components in a physiologically acceptable medium: (a) pinanediol; and (b) at least one soluble collagen, The mass ratio of component (a) to component (b) is (6-20):

1.

2. The skin care composition according to claim 1, wherein The mass ratio of component (a) to component (b) is (9-19):

1.

3. The skin care composition according to claim 1 or 2, wherein The component (b) includes type III collagen.

4. Use of the skin care composition according to any one of claims 1 to 3 for increasing the expression of type I collagen in the skin.

5. Use of the skin care composition according to any one of claims 1 to 3 for improving skin elasticity and firmness.

6. Use of the skin care composition according to any one of claims 1 to 3 in resisting and / or alleviating skin aging.

7. Use of a combination of pinanediol and at least one soluble collagen for increasing the expression of type I collagen in the skin.

8. Use of a combination of pinanediol and at least one soluble collagen for improving skin elasticity and firmness.

9. Use of a combination of pinanediol and at least one soluble collagen in resisting and / or alleviating skin aging.

10. The use according to any one of claims 7 to 9, wherein The mass ratio of the pinanediol to the at least one soluble collagen is (6-20):1.

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