LIQUID CRYSTAL COMPOSITION INCLUDING CERAMIDE AND METHOD OF PREPARING THE SAME

Abstract

Provided a liquid crystal composition including ceramide, a method of preparing the same, and a cosmetic composition including the same. According to a liquid crystal composition including various types of ceramide of an aspect and a method of preparing the same, the liquid crystal composition not only has high liquid crystal stability that liquid crystal is maintained even after 4 weeks at a high temperature, but also activities of increasing an amount of ceramide, increasing an expression level of aquaporin 3, increasing an amount of filaggrin, increasing an expression level of hyaluronic acid, increasing an expression level of loricrin, increasing an expression level of involucrin, and increasing a thickness of the skin, resulting in effects usefully available for amelioration of the skin conditions.

Claims

1. Liquid crystal composition: a water phase; and an oil phase comprising a skin lipid, an emulsifier, and oil, wherein the skin lipid comprises ceramide, cholesterol or a derivative thereof, and a fatty acid.

2. The liquid crystal composition of claim 1, wherein the skin lipid comprises in an amount in a range of about 0.1 wt % to about 10 wt % based on the total weight of the composition.

3. The liquid crystal composition of claim 1, wherein the emulsifier comprises in an amount in a range of about 1 wt % to about 20 wt % based on the total weight of the composition.

4. The liquid crystal composition of claim 1, wherein the oil is comprised in an amount in a range of about 5 wt % to about 30 wt % based on the total weight of the composition.

5. The liquid crystal composition of claim 1, wherein the skin lipid comprises the ceramide, the cholesterol or a derivative thereof, and the fatty acid at a molar ratio of 1:0.5 to 2:0.5 to 2.

6. The liquid crystal composition of claim 1, wherein the ceramide comprises natural ceramide, synthetic ceramide, or pseudo ceramide.

7. The liquid crystal composition of claim 1, wherein the ceramide comprises any one selected from the group consisting of Ceramide 1 (Ceramide EOP), Ceramide 2 (Ceramide NS), Ceramide 3 (Ceramide NP), Ceramide 4 (Ceramide EOH), Ceramide 5 (Ceramide AS), Ceramide 6 (Ceramide AP), Ceramide 7 (Ceramide AH), Ceramide 8 (Ceramide NH), Ceramide 9 (Ceramide EOS), Ceramide 10 (Ceramide Nds), Ceramide 11 (Ceramide Ads), Ceramide 12 (Ceramide EOds), and a combination thereof.

8. The liquid crystal composition of claim 1, wherein the ceramide is hybrid ceramides in which Ceramide3 (Ceramide NP) and Ceramide 6 (Ceramide AP) are mixed at a weight ratio in a range of about 5:1 to about 1:1.

9. The liquid crystal composition of claim 8, wherein the ceramide is hybrid ceramides further comprising Ceramide 9 (Ceramide EOs).

10. The liquid crystal composition of claim 8, wherein the ceramide is hybrid ceramides further comprising two or more ceramides selected from Ceramide 9 (Ceramide EOS), Ceramide 10 (Ceramide Nds), and Ceramide 11 (Ceramide Ads).

11. The liquid crystal composition of claim 1, further comprising any one selected from sphingosine, phytosphingosine, tetraacetylphytosphingosine, and spinganine.

12. The liquid crystal composition of claim 1, the emulsifier has hydrophile-lipophile balance (HLB) in a range of about 3 to about 9.

13. The liquid crystal composition of claim 1, wherein the liquid crystal composition has a hexagonal system crystal structure.

14. The liquid crystal composition of claim 1, wherein the liquid crystal composition has an emulsion formulation.

15. The liquid crystal composition of claim 1, wherein the liquid crystal composition has emulsion particles having an average particle size in a range of about 1 ?m to about 50 ?m.

16. The liquid crystal composition of claim 1, wherein the liquid crystal composition is for ameliorating skin conditions.

17. The liquid crystal composition of claim 16, wherein the amelioration of skin conditions comprises skin barrier strengthening, amelioration of skin aging, amelioration of skin wrinkles, amelioration of skin elasticity, skin regeneration, or skin whitening.

18. A cosmetic composition comprising the liquid crystal composition of any one of claim 1.

19. The cosmetic composition of claim 18, further comprising at least one selected from a stabilizer, a surfactant, oil, a chelating agent, polyol, a preservative, and a thickener.

20. The cosmetic composition of claim 18, wherein the cosmetic composition has a formulation selected from a toner, a lotion, a cream, an essence, a mask pack, a foundation, a soap, a cleansing product, a body moisturizer, a hair product, and a cleanser.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0069] The above and other aspects, features, and advantages of certain embodiments of the disclosure will be more apparent from the following description taken in conjunction with the accompanying drawings, in which:

[0070] FIG. 1 is an image of a liquid crystal composition according to an embodiment observed with a polarizing microscope;

[0071] FIG. 2 is an image of a liquid crystal composition according to an embodiment for analyzing the stability by using a polarizing microscope;

[0072] FIG. 3 is an image (upper panel) and a graph (lower panel) that show effects of a liquid crystal composition according to an embodiment on the production amount of ceramide in the skin;

[0073] FIG. 4 is an image (upper panel) and a graph (lower panel) that show effects of a liquid crystal composition according to an embodiment on the expression level of aquaporin 3 in the skin;

[0074] FIG. 5 is an image (upper panel) and a graph (lower panel) that show effects of a liquid crystal composition according to an embodiment on the expression level of filaggrin in the skin;

[0075] FIG. 6 is a graph that shows effects of a liquid crystal composition according to an embodiment on the expression level of hyaluronic acid in the skin;

[0076] FIG. 7 is an image (upper panel) and a graph (lower panel) that show effects of a liquid crystal composition according to an embodiment on the expression level of aquaporin 3 in the undifferentiated skin;

[0077] FIG. 8 is an image (upper panel) and a graph (lower panel) that show effects of a liquid crystal composition according to an embodiment on the expression level of loricrin in the undifferentiated skin;

[0078] FIG. 9 is an image (upper panel) and a graph (lower panel) that show effects of a liquid crystal composition according to an embodiment on the expression level of involucrin in the undifferentiated skin; and

[0079] FIG. 10 is an image that shows effects of a liquid crystal composition according to an embodiment on the thickness of undifferentiated skin.

DETAILED DESCRIPTION

[0080] Reference will now be made in detail to embodiments, examples of which are illustrated in the accompanying drawings, wherein like reference numerals refer to like elements throughout. In this regard, the present embodiments may have different forms and should not be construed as being limited to the descriptions set forth herein. Accordingly, the embodiments are merely described below, by referring to the figures, to explain aspects. As used herein, the term and/or includes any and all combinations of one or more of the associated listed items. Expressions such as at least one of, when preceding a list of elements, modify the entire list of elements and do not modify the individual elements of the list.

[0081] Hereinafter, preferable Examples are presented to help understanding of the disclosure. However, the following examples are only presented for easier understanding of the disclosure, and the contents of the disclosure are not limited by the following examples. Various transformations can be applied in the examples, and thus examples are not limited to the following examples, and can be implemented in various forms.

Preparation of Liquid Crystal Composition, Examples 1 to 3 and Comparative Examples

[0082] The ingredients listed in Table 1 below were mixed to prepare liquid crystal comparative Examples of Examples 1 to 3 and Comparative Examples 1 and 2.

[0083] Here, the amounts of the following ingredients all expressed in wt %.

[0084] In detail, to prepare a water phase, xanthan gum was dissolved in water at room temperature. Afterwards, glycerin and 1,2-hexanediol were added to the resultant solution, stirred, and heated at a temperature up to 75? C. to 80? C., so as to prepare a water phase. To prepare an oil phase, an emulsifier (e.g., hydrogenated lecithin, glyceryl monostearate, and cetearyl alcohol), a skin lipid (e.g., cholesterol, stearic acid, ceramide), and an additional sphingolipid (e.g., glucosyl ceramide, tetraacetylphytosphingosine, phytosphingosine) were sequentially added to oil (e.g., hydrogenated polydecene), and completely dissolved at a temperature in a range of about 90? C. to about 100? C. Next, while maintaining the temperature between 75? C. and 85? C., the oil phase was stirred and introduced to the water phase. After the introduction of the oil phase was finished, homo-mixing (3000 rpm) was performed thereon for 5 minutes. Afterwards, by rapid cooling down to 35? C. to 40? C., the liquid crystal composition was prepared.

TABLE-US-00001 TABLE 1 Comparative Comparative Ingredients Example 1 Example 2 Example 3 Example 1 Example 2 Oil Hydrogenated polydecene 7.00 10.00 10.00 7.00 10.00 phase Glyceryl monostearate 3.00 3.50 3.50 3.00 Ceteareth-20 3.00 Cetyl alcohol 3.00 2.00 2.00 3.00 3.00 Hydrogenated lecithin 1.50 1.00 1.00 1.50 0.50 Ceramide NP 0.50 1.00 1.00 0.50 1.00 Ceramide AP 0.10 0.50 0.50 0.10 0.50 Ceramide EOP 0.01 0.01 0.01 Ceramide NG 0.10 Ceramide AG 0.10 Cholesterol 0.30 0.90 1.00 0.30 2.70 Stearic acid 0.30 0.90 1.00 0.30 0.90 Phytosphingosine 0.5 0.5 Tetraacetylphytosphingosine 0.25 0.25 0.25 0.25 0.25 Glucosylceramide 0.001 0.001 0.001 0.001 0.001 Water Purified water 77.049 72.439 72.539 77.049 70.639 phase Glycerin 5.00 5.00 5.00 5.00 5.00 1,2-hexanediol 2.00 2.00 2.00 2.00 2.00 Total 100 100 100 100 100

[0085] In the liquid crystal compositions of Examples 1 to 3, the mass ratio of ceramide:cholesterol:fatty acid was 40:25:25. However, the liquid crystal composition was prepared to include these ingredients at a ratio of 1:1:1 when the mass ratio was converted to a molar ratio.

Experimental Example 1. Analysis of Crystal Structure of Liquid Crystal Composition

[0086] The crystals of the liquid crystal compositions of Examples 1 to 3 and Comparative Examples 1 to 2 were analyzed with a polarizing microscope, and the results are shown in FIG. 1.

[0087] As shown in FIG. 1, it was confirmed that the crystals of the liquid crystal compositions of Examples 1 to 3 showed a homogeneous hexagonal liquid crystal form well, whereas the crystals of the liquid crystal compositions of Comparative Examples 1 and 2 did not form hexagonal liquid crystals well.

Experimental Example 2. Analysis of Stability of Liquid Crystal Composition

[0088] To analyze the stability of the liquid crystal composition, the liquid crystal composition of Example 2 was stored at different temperatures (25? C., 4? C., and 45? C.), and then the presence or absence of liquid crystals was analyzed with a polarizing microscope every week, and the results are shown in FIG. 2.

[0089] As shown in FIG. 2, it was confirmed that the liquid crystals of the liquid crystal composition according to Example was maintained for 4 weeks at all temperatures (25? C., 4? C., and 45? C.).

Experimental Example 3. Confirmation of Ceramide Production Amount Using Artificial Skin

[0090] To confirm the amount of ceramide produced by the liquid crystal composition, the liquid crystal compositions of Examples 1 and 2 were applied to artificial skin (EPI-200-X, MatTek), and after 4 hours, the permeation and production amount of ceramide were confirmed. The amount of ceramide was analyzed by immunofluorescence. Specifically, artificial skin was treated with the liquid crystal compositions of Examples, cultured for 24 hours, and then washed out by using PBS. The resultant artificial skin was put into a cassette and fixed with 10% formalin. After cutting the artificial skin tissue to a thickness of 3 om to prepare a slide, paraffin was melt thereon in a dry oven at a temperature of 60? C. for 30 minutes. To remove the paraffin, each slide was treated with xylene for 15 minutes, 100% ethanol for 5 minutes, 95% ethanol for 5 minutes, and 70% ethanol for 5 minutes, respectively, and then subjected to cement hydration with running water. Antigen retrieval was performed on the resultant slides by heating in a citrate buffer for 10 minutes by using a cooker device, followed by cooling in running water. After treated with 0.5% triton X-100 for 15 minutes, the resultant slides were washed with PBS buffer three times for 5 minutes each. Blocking with 1% BSA was then performed thereon, followed by washing with PBS three times for 5 minutes each.

[0091] The primary antibody was diluted at 1:50 and reacted overnight at 4? C., and citrate buffer was used for antigen retrieval.

[0092] After washing with PBS, the FITC 488 antibody was reacted for 1 hour at room temperature and washed three times with PBS. The slides were fixed by DAPI staining and using a vectashield mounting agent, and then observed under a fluorescence microscope. As a control group, an untreated liquid crystal composition was used. The analysis results are shown in FIG. 3.

[0093] As shown in FIG. 3, it was confirmed that the amount of ceramide produced in the skin was increased according to the application of the liquid crystal composition of Examples.

Experimental Example 4. Confirmation of Expression Level of Aquaporin 3 by Using Artificial Skin

[0094] To confirm the change in the expression level of aquaporin 3 by the liquid crystal composition, the liquid crystal compositions of Examples 1 and 2 were applied to artificial skin (EPI-200-X, MatTek), and after 24 hours, the expression of aquaporin 3 was analyzed in the same manner as in Experimental Example 3 according to immunofluorescence.

[0095] As shown in FIG. 4, it was confirmed that the expression level of aquaporin 3 in the skin was increased according to the application of the liquid crystal composition of Examples.

Experimental Example 5. Confirmation of Expression Level of Filaggrin by Using Artificial Skin

[0096] To confirm the change in the expression level of filaggrin by the liquid crystal composition, the liquid crystal compositions of Examples 1 and 2 were applied to artificial skin (EPI-200-X, MatTek), and after 24 hours, the expression of filaggrin was analyzed in the same manner as in Experimental Example 3 according to immunofluorescence. The results are shown in FIG. 5.

[0097] As shown in FIG. 5, it was confirmed that the expression level of filaggrin in the skin was increased according to the application of the liquid crystal composition of Examples.

Experimental Example 6. Confirmation of Hyaluronic Acid Production Amount Using Artificial Skin

[0098] To confirm the amount of hyaluronic acid produced by the liquid crystal composition, the liquid crystal compositions of Examples 1 and 2 were applied to artificial skin (EPI-200-X, MatTek), and after 6 hours, the amount of hyaluronic acid produced was confirmed. The amount of hyaluronic acid was analyzed by using a Hyaluronan DuoSet ELISA (DY3614-05) according to the instructions of the manufacturer, and the results are shown in FIG. 6.

[0099] As shown in FIG. 6, it was confirmed that, according to the application of the liquid crystal compositions of Examples, the amount of hyaluronic acid produced after 6 hours was increased compared to the control group, and that the amount of hyaluronic acid produced after 24 hours was significantly increased compared to the control group.

[0100] Based on the results above, it was confirmed that the liquid crystal compositions of Examples had effects on the amelioration of the skin conditions, for example, strengthening the skin barrier and moisturizing the skin.

Experimental Example 7. Confirmation of Expression Level of Aquaporin 3 by Liquid Crystal Composition

[0101] To confirm the change in the skin moisturizing ability, specifically the expression level of aquaporin 3, by the liquid crystal composition, the liquid crystal compositions of Example 2 and Comparative Example 1 were applied to undifferentiated artificial skin (EPI-201, MatTek), and after 24 hours, the expression level of aquaporin 3 was analyzed in the same manner as in Experimental Example 3 according to immunofluorescence. The results are shown in FIG. 7.

[0102] As shown in FIG. 7, as the skin differentiation process proceeded, the expression level of aquaporin 3 was increased regardless of the presence or absence of the liquid crystal composition of Example. However, it was confirmed that the expression level of aquaporin 3 in Example 2 was significantly improved compared to Comparative Example 1 having a non-liquid crystal structure.

Experimental Example 8. Confirmation of Expression Level of Loricrin by Liquid Crystal Composition

[0103] To confirm the change in the skin differentiation factors, specifically the expression level of loricrin, by the liquid crystal composition, the liquid crystal compositions of Example 2 and Comparative Example 1 were applied to undifferentiated artificial skin (EPI-201, MatTek), and after 24 hours, the expression level of loricrin was analyzed in the same manner as in Experimental Example 3 according to immunofluorescence. The results are shown in FIG. 8.

[0104] As shown in FIG. 8, it was confirmed that the expression level of loricrin in the skin was increased according to the application of the liquid crystal composition of Examples. In addition, it was confirmed that the expression level of loricrin in the liquid crystal structure of Example was significantly greater than the expression level of loricrin in the non-liquid crystal structure of Comparative Example.

Experimental Example 9. Confirmation of Expression Level of Involucrin by Liquid Crystal Composition

[0105] To confirm the change in the skin differentiation factors, specifically the expression level of involucrin, by the liquid crystal composition, the liquid crystal compositions of Example 2 and Comparative Example 1 were applied to undifferentiated artificial skin (EPI-201, MatTek), and after 24 hours, the expression level of involucrin was analyzed in the same manner as in Experimental Example 3 according to immunofluorescence. The results are shown in FIG. 9.

[0106] As shown in FIG. 9, it was confirmed that the expression level of involucrin in the skin was increased according to the application of the liquid crystal composition of Examples. In addition, it was confirmed that the expression level of involucrin in the liquid crystal structure of Example was significantly greater than the expression level of involucrin in the non-liquid crystal structure of Comparative Example.

Experimental Example 10. Confirmation of Skin Thickness and Differentiation Degree by Liquid Crystal Composition

[0107] To confirm the difference in skin differentiation and skin barrier thickness by the liquid crystal composition, the liquid crystal compositions of Example 2 and Comparative Example 1 were applied to undifferentiated artificial skin (EPI-201, MatTek), and after 24 hours, the skin thickness was measured. The skin thickness was confirmed through H&E staining. In detail, in the same manner as in Experimental Example 3, a paraffin block was prepared, and a slide was prepared by cutting the artificial skin tissue to a thickness of 3 om to remove paraffin, and then paraffin was melt thereon in a dry oven at a temperature of 60? C. for 30 minutes. The slide was immersed in xylene, 100% ethanol, 85% ethanol, 70% ethanol, and water in this stated order, followed by staining with hematoxylin for 10 minutes. After washing with running water, the resultant slide was immersed in 1% HCl and 1% ammonia in this stated order. After staining with eosin for 2 minutes, dehydration was performed thereon, and the resultant slide was mounted, and the results obtained by observation under a microscope are shown in FIG. 10.

[0108] As shown in FIG. 10, it was confirmed that the skin thickness and differentiation of skin cells were increased according to the application of the liquid crystal composition of Examples. In addition, it was confirmed that the skin thickness in the liquid crystal structure of Example was significantly greater than the skin thickness in the non-liquid crystal structure of Comparative Example.

[0109] Based on the results above, it was confirmed that the liquid crystal compositions of Examples had effects of improving the condition of undifferentiated skin, for example, strengthening skin barrier and inducing skin differentiation.

[0110] According to a liquid crystal composition including various types of ceramide of an aspect and a method of preparing the same, the liquid crystal composition not only has high liquid crystal stability that liquid crystal is maintained even after 4 weeks at a high temperature, but also activities of increasing an amount of ceramide, increasing an expression level of aquaporin 3, increasing an amount of filaggrin, increasing an expression level of hyaluronic acid, increasing an expression level of loricrin, increasing an expression level of involucrin, and increasing a thickness of the skin, resulting in effects usefully available for amelioration of the skin conditions.

[0111] It should be understood that embodiments described herein should be considered in a descriptive sense only and not for purposes of limitation. Descriptions of features or aspects within each embodiment should typically be considered as available for other similar features or aspects in other embodiments. While one or more embodiments have been described with reference to the figures, it will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope as defined by the following claims.