Pseudo-ceramide complex derived from citron-seed oil, and cosmetic composition comprising same for moisturizing skin and enhancing skin barrier

Abstract

The present invention relates to a cosmetic composition containing pseudo-ceramide derived from vegetable oil as an active ingredient and, specifically, to a cosmetic composition containing pseudo-ceramide derived from citron seed oil as an active ingredient, which does not irritate the skin, and has an excellent skin moisturizing effect and barrier strengthening function.

Description

A ceramide-like complex derived from citron seed oil, and a cosmetic composition for skin moisturization and skin barrier strengthening containing the same.

[0001] The present invention relates to a ceramide-like complex derived from vegetable oil, and a cosmetic composition containing the same for moisturizing the skin and strengthening the skin barrier.

[0002] The stratum corneum of the skin's epidermis is composed of a brick-like multilayer structure, and the lamellar structure, consisting of lipids between keratinocytes, is bound by ceramide, cholesterol, and free fatty acids. Among these, ceramide is the main component of epidermal lipids, accounting for approximately 40–65%, and plays a central role in maintaining moisture and the barrier function of the stratum corneum. When the ceramide content in the stratum corneum decreases, the protective barrier function of the stratum corneum is reduced, and various skin diseases such as atopic dermatitis worsen, leading to skin aging; however, it has been reported that the skin can be restored to a normal state by supplementing ceramide from the outside.

[0003] These ceramides have been extracted from plants, animals, or microorganisms and utilized as natural ceramides; however, commercialization is difficult due to challenges such as the difficulty of mass production, excessively high production costs, and very low solubility in various solvents used in cosmetics and raw materials used in cleansing products. Consequently, active research is being conducted on ceramide-like substitutes that have a structure similar to natural ceramides, high solubility in various solvents, and similar efficacy.

[0004] Meanwhile, upcycling materials, a recent trend in the cosmetics industry, are gaining attention. Upcycling is a process that assigns new value to by-products and waste materials that are no longer in use, going beyond the concept of recycling. In Korea, while approximately 10,000 tons of yuzu are processed annually, yuzu seeds, a by-product, account for about 20% (2,000 tons) of the raw material. Although some farms use them as fertilizer, most are currently disposed of, causing regional problems (Non-patent Literature 1: Ko, Eun Ah, et al., Journal of the Society of Cosmetic Scientists of Korea 46.3 (2020): 283-294; thereby, the entire content of Non-patent Literature 1 is incorporated and adopted as the prior art content of this specification).

[0005] Therefore, there is a demand for various upcycling technologies that utilize yuzu seeds, which are currently being discarded in large quantities.

[0006] Accordingly, the inventors confirmed that when pseudo-ceramide is produced from yuzu seed oil, a pseudo-ceramide complex can be obtained more efficiently and economically than with other vegetable oils, and furthermore, the obtained pseudo-ceramide complex derived from yuzu seed oil has high stability and is non-irritating to the skin, while also possessing excellent skin moisturizing effects, skin inflammation alleviation effects, and skin barrier strengthening effects, and thus completed the present invention.

[0007] (Non-patent Document 1) Ko, Eun Ah, et al., Journal of the Society of Cosmetic Scientists of Korea 46.3 (2020): 283-294

[0008] As mentioned above, there is a need for technology to utilize yuzu seeds, which are currently being discarded in large quantities. Accordingly, yuzu seed oil was obtained from these discarded yuzu seeds, and a pseudo-ceramide complex was manufactured using it.

[0009] The present invention aims to solve the problems of the aforementioned prior art and to provide a cosmetic composition containing a pseudo-ceramide complex derived from yuzu seed oil.

[0010] In order to solve the problems of the aforementioned prior art, the present invention

[0011] A step of reacting vegetable oil, amino alcohol, and lipase by adding them to an organic solvent; and

[0012] The method sequentially includes the step of adding additional unsaturated fatty acids and lipase to the above reactants and reacting them again, wherein

[0013] The present invention provides a method for manufacturing a pseudo-ceramide complex characterized in that the vegetable oil is yuzu seed oil.

[0014] In addition, the present invention provides a method for manufacturing a pseudo-ceramide complex characterized in that the yuzu seed oil contains at least 35% oleic acid and at least 20% palmitic acid.

[0015] In addition, the present invention provides a method for manufacturing a pseudo-ceramide complex characterized in that the yuzu seed oil has a saponification value of 180 to 210, an iodine value of 90 to 110, and an acid value of 0.3 to 0.6.

[0016] In addition, the present invention provides a method for preparing a pseudo-ceramide complex characterized in that the additional unsaturated fatty acid is oleic acid.

[0017] In addition, the present invention provides a method for producing a pseudo-ceramide characterized in that the amino alcohol is 3-amino-1-propanol.

[0018] In addition, the present invention provides a method for manufacturing a pseudo-ceramide complex characterized in that the organic solvent is hexane.

[0019] In addition, the present invention provides a method for preparing a pseudo-ceramide complex, characterized in that the lipase is a CAL-B enzyme or a variant thereof.

[0020] In addition, the present invention provides a citron seed oil-derived ceramide complex prepared according to the manufacturing method of the present invention.

[0021] In addition, the present invention provides a citron seed oil-derived ceramide complex characterized by being represented by the following chemical formula 1.

[0022] [Chemical Formula 1]

[0023]

[0024] Here, R in the above chemical formula 1 , or R 2 Each is an acyl residue derived from the fatty acid or oleic acid of citron seed oil, independently.

[0025] In addition, the present invention provides a citron seed oil-derived pseudo-ceramide complex characterized by comprising one or more compounds represented by the following chemical formulas 2 to 6.

[0026] [Chemical Formula 2]

[0027]

[0028] [Chemical Formula 3]

[0029]

[0030] [Chemical Formula 4]

[0031]

[0032] [Chemical Formula 5]

[0033]

[0034] [Chemical Formula 6]

[0035]

[0036] In addition, the present invention provides a citron seed oil-derived ceramide complex characterized by including 35 to 40 weight percent of the compound of Formula 2 based on the total weight of the composition.

[0037] In addition, the present invention provides a citron seed oil-derived ceramide complex characterized by including 35 to 40 weight percent of the compound of Formula 2 based on the total weight of the composition.

[0038] In addition, a cosmetic composition is provided containing 0.001 to 10% by weight of the yuzu seed oil-derived ceramide complex of the present invention based on the total weight of the cosmetic composition.

[0039] In addition, in the cosmetic composition of the present invention, the composition comprises: caprylic / capric triglyceride 6-10 wt%, coco-caprylate / caprate 3-7 wt%, 2,3-butanediol 3-6 wt%, glycerin 1-5 wt%, pentaerythrityl distearate 1-3 wt%, cetearyl glucoside 0.5-2.5 wt%, cetearyl alcohol 0.5-2.5 wt%, shea butter 0.5-2.5 wt%, vinyl dimethicone 0.5-2.5 wt%, 1,2-hexanediol 1-2 wt%, stearyl alcohol 0.1-0.5 wt%, stearic acid 0.1-0.3 wt%, tromethamine 0.1-0.4 wt%, and sodium stearoyl glutamate. A cosmetic composition is provided that is characterized by a cream formulation comprising 0.1~0.3 wt%, 0.2~0.4 wt% carbomer, 0.01~0.03 wt% disodium EDTA, 0.02~0.04 wt% fragrance, and 65~75 wt% purified water.

[0040] In addition, the present invention provides a cosmetic composition characterized in that, in the above-described cosmetic composition, the cosmetic composition is for strengthening the skin barrier, for alleviating skin inflammation, or for moisturizing the skin.

[0041] By utilizing discarded yuzu seed oil, it is possible to manufacture pseudo-ceramide complexes with a high yield compared to other vegetable oils, in addition to achieving an upcycling effect. Furthermore, since the obtained pseudo-ceramide complex derived from yuzu seed oil exhibits excellent skin barrier strengthening, moisturizing, and skin inflammation alleviation effects, it can be usefully applied as a cosmetic product for moisturizing, skin regeneration, skin elasticity improvement, and atopic dermatitis improvement.

[0042] Figure 1 is a diagram showing a reaction scheme for producing pseudo-ceramide from citron seed oil according to the present invention.

[0043] Figure 2 is a graph of the gas chromatography analysis results.

[0044] Figure 3 is a photograph of test results showing the degree of dissolution of the pseudo-ceramide complex of Example 1 of the present invention and Ceramide NP in each of four solvents, MCT Oil (caprylic / capric triglyceride), Dimethicone, 1,3 BG (1,3-butylene Glycol), and Greendiol™ (2,3-butanediol), at concentrations of 1% and 3%.

[0045] Figure 4 is a graph showing the test results for the amount of hyaluronic acid produced by the pseudo-ceramide complex of Example 1 of the present invention.

[0046] Figure 5 is a graph showing the results of a test on the production of Filaggrin (FLG) of the pseudo-ceramide complex of Example 1 of the present invention.

[0047] Figure 6 is a graph showing the results of a test on the inhibition of Prostaglandin E2 (PGE2) production by the pseudo-ceramide complex of Example 1 of the present invention.

[0048] The present invention will be described in detail below.

[0049]

[0050] One aspect of the present invention is,

[0051] A step of reacting vegetable oil, amino alcohol, and lipase by adding them to an organic solvent; and

[0052] The method sequentially includes the step of adding additional unsaturated fatty acids and lipase to the above reactants and reacting them again, wherein

[0053] The present invention relates to a method for manufacturing a pseudo-ceramide complex characterized in that the above-mentioned vegetable oil is yuzu seed oil. Through various experiments, the inventors discovered that among various vegetable oils, yuzu seed oil synthesizes pseudo-ceramide with the highest yield when synthesizing a pseudo-ceramide complex. They specifically selected yuzu seed oil, which enables the synthesis of pseudo-ceramide with high purity and high yield, to complete the present invention. That is, while other vegetable oils using the same method generally show a yield in the 80% range, it was discovered that yuzu seed oil specifically exhibits a high yield of over 90%, and thus it was selected as a particularly desirable oil for the production of pseudo-ceramide. Furthermore, by utilizing discarded yuzu seed oil to manufacture a high-value pseudo-ceramide that has higher stability than the oil itself and possesses effects such as strengthening skin barrier function, moisturizing the skin, and alleviating skin inflammation, an upcycling effect can be achieved. A more detailed understanding of this can be obtained through the examples and experimental examples described below.

[0054] In one embodiment, the citron seed oil (Citrus junos seed oil) may be a fatty oil obtained by cold-pressing the seeds of citron from Goheung, Jeollanam-do without using an organic solvent, and the citron seed oil is desirable for smoothness and spreadability. Citron seed oil mainly contains fatty acids such as oleic acid (C18:1) and linoleic acid (C18:2), and has antioxidant, anti-inflammatory, and anti-allergic effects, and has excellent skin moisturizing properties, which helps improve atopic dermatitis.

[0055] In particular, it is preferable to use the above yuzu seed oil containing at least 35% oleic acid and at least 20% palmitic acid.

[0056] It has been confirmed that using yuzu seed oil containing oleic acid and palmitic acid within the above range is relatively suitable for the pseudo-ceramide manufacturing process and that it is possible to manufacture pseudo-ceramide with excellent effects. In this regard, the oleic acid and palmitic acid may serve as indicator components that serve as a standard for the quality of the yuzu seed oil of the present invention, and at the same time, as functional components that provide beneficial effects to the skin.

[0057] In other words, oleic acid is a monounsaturated fatty acid that is not destroyed by heat, and palmitic acid is a stable saturated fatty acid, making it not only desirable as an indicator component but also capable of providing moisturizing effects in terms of skin benefits.

[0058] Similarly, ceramides derived from oleic acid and palmitic acid can also serve as quality standards for citron seed oil-derived ceramide complexes or as functional ingredients that provide beneficial effects to the skin. For example, ceramide components derived from oleic acid or palmitic acid can help improve tissue density and help strengthen skin barrier function. A more detailed understanding of this can be achieved through the examples and experimental examples described below.

[0059] The above yuzu seed oil preferably has a saponification value of 180 to 210 (mg / g), an iodine value of 90 to 110 (g / 100g), and an acid value of 0.3 to 0.6 (mg / g). The saponification value is the number of mg of KOH required to saponify the oil and tends to decrease due to rancidity or oxidation of the oil; the iodine value is a measure of the degree of unsaturation (degree of double bonds) and tends to decrease during oxidation or rancidity; and the acid value is a measure of free fatty acids and may increase during extraction, oil processing, or storage.

[0060] Although there are currently no established raw material standards or specifications for the yuzu seed oil in use, it has been confirmed that oils with saponification, iodine, and acid values ​​within this approximate range are suitable for the smooth production of pseudo-ceramides without issues of oxidation or rancidity. A more detailed understanding of this can be achieved through the examples and experimental examples described later.

[0061] As described above, the oleic acid and palmitic acid content, or the saponification value, iodine value, acid value, etc., can be referred to as raw material standards for yuzu seed oil that is desirable for the production of pseudo-ceramide.

[0062] It is preferable that the additional unsaturated fatty acid mentioned above be oleic acid. Oleic acid is a monounsaturated fatty acid that is not destroyed by heat and is a component found in large quantities in some vegetable oils, such as camellia oil.

[0063] The above amino alcohol can be exemplified by various amino alcohols such as 3-amino-1,2-propanediol, 3-amino-1-propanol, 2-amino-1-butanol, 1-amino-2-propanol, 2-amino-2-ethyl-1,3-propanediol, 2-amino-2-ethyl-1,3-propanediol, 2-amino-1,3-propanediol, etc., but it is most preferable to be 3-amino-1-propanol (3-amino-1-propanol) in terms of production efficiency through the amination reaction with yuzu seed oil and in terms of the physical properties and effects of the generated pseudo-ceramide.

[0064] The above organic solvent may be any organic solvent widely known in the art, and non-limiting examples include hexane and t-amyl alcohol (TAA). The above organic solvent may be a known solvent used alone, or a known solvent may be used in combination.

[0065] The above lipase is an enzyme that breaks down fat into fatty acids and monoglycerides, and is also referred to as a lipolytic enzyme or lipase. While commercially available lipases may be used without limitation, CAL-B enzymes or their variants may be used, and examples include CalB1422-L278L, CalB1422-L278A, L278G, L278H, etc.

[0066]

[0067] Another aspect of the present invention is,

[0068] The present invention relates to a citron seed oil-derived ceramide produced according to the manufacturing method of the present invention described above, and may be a citron seed oil-derived ceramide characterized by being represented by the following chemical formula 1.

[0069] [Chemical Formula 1]

[0070]

[0071] Here, R in the above chemical formula 1 , or R 2 Each is an acyl residue derived from the fatty acid or oleic acid of citron seed oil, independently.

[0072] The compound of Chemical Formula 1 above may include various compounds such as Chemical Formulas 2 to 6 below, depending on the type of fatty acid derived from yuzu seed oil.

[0073] [Chemical Formula 2]

[0074]

[0075] [Chemical Formula 3]

[0076]

[0077] [Chemical Formula 4]

[0078]

[0079] [Chemical Formula 5]

[0080]

[0081] [Chemical Formula 6]

[0082]

[0083]

[0084] Also, another aspect of the present invention is,

[0085] The present invention relates to a cosmetic composition containing a citron seed oil-derived ceramide complex as described above.

[0086] The yuzu seed oil-derived pseudo-ceramide complex of the present invention may contain approximately 0.001 to 10 weight% based on the total weight of the composition, but preferably 0.01 to 10 weight%, more preferably 0.1 to 7 weight%, even more preferably 0.3 to 5 weight%, and most preferably 0.5 to 3 weight%. Below the above range, the pseudo-ceramide content may be too low, which may result in insufficient effects from its inclusion; above the above range, there may be concerns that it may be difficult to manufacture into a cosmetic composition due to increased viscosity or reduced compatibility, as well as concerns that formulation stability may be compromised due to reduced solubility. For reference, commonly used ceramide NPs present significant difficulties in formulation application when used at such high concentrations; therefore, the ability to use them in cosmetic compositions at high concentrations may be a major advantage of the yuzu seed oil-derived pseudo-ceramide complex.

[0087] Such compositions may include, for example, vehicles permitted in commonly used ordinary cosmetics, such as diluents, dispersants, gelling agents, carriers for ceramides, solidifying agents, gums, resins, tension activators, solvents, fillers, such as rice starch, pigments, preservatives, essential oils, antioxidants, coloring agents, pearl fibers, fragrances, deodorizers, pH adjusters or neutralizing agents, and viscosity enhancers.

[0088] In addition, the composition exists in forms suitable for application to the skin and foreskin, such as gels, lotions, particularly capillary lotions and varnishes, particularly O / W or W / O type emulsions or dispersants, creams, particularly mascara creams, ointments, milks, foams, sticks, particularly lip creams and lipsticks.

[0089] The compound obtained by the method according to the present invention may be formulated in a manner similar to that described in the prior art. Without limitation, it may be useful to convert it into examples of the listed formulations.

[0090] In the form of an emulsion, the composition according to the present invention contains an emulsifier and an auxiliary emulsifier as used by those skilled in the art. Suitable examples of emulsifiers and auxiliary emulsifiers are fatty acid and polyol esters, such as glyceryl stearate, and fatty acid and polyethylene glycol esters, such as PEG-20 stearate. Advantageously, the concentration of the emulsifier and auxiliary emulsifier is 0.3 to 30% w / w, preferably 0.5 to 5%.

[0091] In a specific embodiment of the present invention, the cosmetic composition comprises 0.4 to 0.6 wt%, caprylic / capric triglyceride 6 to 10 wt%, coco-caprylate / caprate 3 to 7 wt%, 2,3-butanediol 3 to 6 wt%, glycerin 1 to 5 wt%, pentaerythrityl distearate 1 to 3 wt%, cetearyl glucoside 0.5 to 2.5 wt%, cetearyl alcohol 0.5 to 2.5 wt%, shea butter 0.5 to 2.5 wt%, vinyl dimethicone 0.5 to 2.5 wt%, 1,2-hexanediol 1 to 2 wt%, stearyl alcohol 0.1 to 0.5 wt%, stearic acid 0.1 to 0.3 wt%, and tromethamine 0.1 to 0.4 wt%. It is preferable to formulate a cream formulation with 0.1~0.3% by weight of sodium stearoyl glutamate, 0.2~0.4% by weight of carbomer, 0.01~0.03% by weight of disodium EDTA, 0.02~0.04% by weight of fragrance, and 65~75% by weight of purified water.

[0092] Based on the total weight of the cream formulation, the cosmetic composition comprises 0.5 wt%, caprylic / capric triglyceride 8 wt%, coco-caprylate / caprate 5 wt%, 2,3-butanediol 4.5 wt%, glycerin 3 wt%, pentaerythrityl distearate 1.8 wt%, cetearyl glucoside 1.25 wt%, cetearyl alcohol 1.25 wt%, shea butter 1.25 wt%, vinyl dimethicone 1.25 wt%, 1,2-hexanediol 1.5 wt%, stearyl alcohol 0.3 wt%, stearic acid 0.2 wt%, tromethamine 0.28 wt%, sodium stearoyl glutamate 0.2 wt%, carbomer 0.3 wt%, and disodium EDTA. A cream formulation composition formed by mixing in a ratio of 0.02% by weight, 0.03% by weight of fragrance, and 69.37% by weight of purified water is even more preferable.

[0093] The above cosmetic composition can be used for moisturizing, skin regeneration, skin elasticity improvement, and atopic dermatitis improvement.

[0094] The above cosmetic composition can be used for strengthening the skin barrier or for moisturizing, and its efficacy has been confirmed. More specific efficacy and effects may be demonstrated through the examples and experimental examples described later.

[0095]

[0096]

[0097] The present invention will be explained in more detail below through examples and test examples. However, it should be clarified that the following examples and test examples are for the purpose of explaining the invention in detail only and are not intended to interpret the scope of rights.

[0098]

[0099] Examples

[0100] Example 1: Preparation of Citron Seed Oil-Derived Ceramide-like

[0101] Yuzu seed oil extract

[0102] The yuzu used was seeds from yuzu produced in Goheung, Jeollanam-do.

[0103] First, foreign substances contained in the yuzu seeds stored at low temperature were removed using a mesh, and the seeds were washed 1 to 3 times by immersion in water. Then, the washed yuzu seeds were dried at 25 to 30°C, and the foreign substances were separated. Next, the yuzu seeds were cold-pressed using an oil press at a temperature of 20°C to produce pressed oil. Then, 5% activated carbon and 4% deoxidized clay were added to the pressed oil, and the mixture was filtered using a filter press. 0.05% of the antioxidant DL-alpha tocopherol was added to the filtered pressed oil, and the mixture was filtered through a 1+0.45 μm filter to produce yuzu seed oil.

[0104]

[0105] Yuzu Seed Oil Specification Analysis

[0106] To analyze the fatty acids of citron seed oil, a gas chromatography system equipped with an automatic injector and a flame ionization detector (FID) were used.

[0107] Ingredient g / 100g (%)Oleic acid37.05Linoleic acid33.53Palmitic acid20.25Stearic acid3.84Linolenic acid1.82Palmitoleic acid0.70Arachidic acid0.31Heptadecanoic acid0.11

[0108] The results were as shown in Table 1 above, and it was confirmed that the yuzu seed oil contained the highest amounts of unsaturated fatty acids, oleic acid and linoleic acid, at 37.05% and 33.53%, respectively, followed by the saturated fatty acid palmitic acid at 20.25%. Meanwhile, the yuzu seed oil was transparent with a light yellow color and had a slight peculiar odor. Its acid value was approximately 0.38 (mg / g), iodine value 104.79 (g / 100g), saponification value 187 (mg / g), and specific gravity 0.89, and heavy metals such as lead and arsenic were not detected.

[0109]

[0110] Preparation of a ceramide complex derived from yuzu seed oil

[0111] In the preparation of pseudoceramide from citron seed oil, as shown in Fig. 1, the amidation of primary fatty acids must precede, and the secondary fatty acids are then esterified to the fatty acid amide produced therefrom to form the final pseudoceramide.

[0112] First, for the first-stage reaction, yuzu seed oil and 3-amino-1-propanol were mixed at a substrate molar ratio of 1.5:1, and the first-stage reaction was carried out using a lipase enzyme. Next, a second-stage reaction was conducted to induce a re-reaction of unreacted fatty acid amides remaining from the first stage and to increase the conversion yield of pseudoceramide. After removing the water and unreacted amino alcohols generated in the first stage reaction through distillation, pseudoceramide was recovered in the first stage. In the second stage reaction, to further convert the unreacted fatty acid amides remaining from the first stage into pseudoceramide, the amount of residual fatty acid amides was quantified, and an equal molar amount of oleic acid was added. The substrate for the second stage reaction was dissolved in hexane, a lipase enzyme was added, and the reaction was carried out at 50°C.

[0113] Specifically, a primary reaction substrate was prepared by mixing amino alcohol yuzu seed oil, 3-amino-1-propanol, and a solvent (hexane:TAA 1:1 (v / v)), and a lipase enzyme was added to this mixture, followed by a primary reaction at 50°C. After removing the enzyme from the reaction mixture, water and residual amino alcohol were removed by distillation, and a secondary reaction was carried out. As a secondary reaction substrate, oleic acid was added to the residual fatty acid amide in an amount equal to the molar ratio of the residual fatty acid amide, and hexane and a lipase enzyme were added as a secondary solvent, followed by a secondary reaction at 50°C for 60 hours. Subsequently, the enzyme was removed from the reaction mixture, followed by distillation for 2 hours. Additionally, decolorization and deodorization processes were performed to produce pseudoceramide.

[0114] As a result, as shown in Figure 2, the production of a high-purity pseudoceramide complex derived from yuzu seed oil was confirmed with high yield (approximately 93%). For reference, Figure 2 is a graph of the results of analyzing the pseudoceramide complex using gas chromatography (7890A, Agilent Technologies, Santa Clara, CA, USA) equipped with an HP-5 capillary column (30 m length x 0.320 mm diameter, 0.25 μm film thickness, Hewlett Packard, USA).

[0115]

[0116] Example 2: Preparation of a cream formulation using a cosmetic composition containing a pseudo-ceramide complex

[0117] To select appropriate solvents for the pseudo-ceramide complex prepared in Example 1 above and commercially available ceramide NP, the solubility was evaluated by dissolving the complex in each of four solvents—MCT Oil (caprylic / capric triglyceride), dimethicone, 1,3 BG (1,3-butylene Glycol), and Greendiol™ (2,3-butanediol)—at concentrations of 1% and 3% (test conditions: temperature 80℃, heating for 1 hour). The results are shown in Figure 3.

[0118] As shown in Figure 3, both the pseudo-ceramide complex prepared in Example 1 and the commercially available ceramide NP exhibited the best solubility in MCT Oil; however, not only did Example 1 exhibit better solubility, but the pseudo-ceramide complex prepared in Example 1 also showed better solubility compared to the commercially available ceramide NP across all other solvents.

[0119] Meanwhile, in order to prepare a cosmetic composition containing the pseudo-ceramide complex prepared in Example 1 above, the caprylic / capric triglyceride (MCT Oil) with the best solubility in the solvent test was selected, and a cream formulation was prepared with the composition of Table 2 below.

[0120] Composition Example 2 Example 3 Example 4 Comparative Example 1 Comparative Example 2 Comparative Example 3 Ceramide-like (Example 1) 0.513-Ceramide NP*---0.513 Caprylic / Capric Triglyceride 8.00 Coco-Caprylate / Caprate 5.00 2,3-Butanediol 4.50 Glycerin 3.00 Pentaerythrityl Distearate 1.80 Cetearyl Glucoside 1.25 Cetearyl Alcohol 1.25 Shea Butter 1.25 Vinyl Dimethicone 1.25 1,2-Hexanediol 1.50 Stearyl Alcohol 0.30 Stearic Acid 0.20 Tromethamine 0.28 Sodium Stearoyl Glutamate 0.20 Carbomer 0.30 Disodium EDTA 0.02 Fragrance 0.03 Purified Water 69.37 (Unit: Weight%)* Commercially available Ceramide NP was purchased and used

[0121]

[0122] Test example

[0123] Test Example 1: Confirmation of skin moisturizing effect of pseudo-ceramide complex

[0124] The skin moisturizing effect of the cosmetic composition containing the pseudo-ceramide complex of Example 1 above was confirmed as follows.

[0125] HaCaT Cell 1.0 X 10 5 Cells / ml were inoculated into a 6-well plate using DMEM (Dulbecco's Modified Eagle Medium) and cultured for 24 hours. The cosmetic composition containing pseudo-ceramide and ceramide NP were each added to fresh medium at 20 μg / mL, and a positive control was added at 10 mM of N-Acetylglucosamine, which was cultured for 24 hours. After collecting the supernatant from the cultured cells, an ELISA assay was performed using the Hyaluronan Quantikine ELISA kit (R&D System, USA) according to the manufacturer's manual to measure the amount of hyaluronic acid produced. The measurement results are shown in Figure 4.

[0126] As confirmed in Figure 4, the amount of hyaluronic acid produced by the pseudo-ceramide cosmetic composition was found to increase by approximately 7.09%, and it was found to be statistically significantly higher than that of commercial ceramide, ceramide NP.

[0127]

[0128] Test Example 2: Confirmation of Filaggrin (FLG) generation effect of pseudo-ceramide complex

[0129] The skin moisturizing effect of the pseudo-ceramide complex of Example 1 above was confirmed as follows.

[0130] HaCaT Cell 1.0 X 10 5 Cells / ml were inoculated into a 6-well plate using DMEM (Dulbecco's Modified Eagle Medium) and cultured for 24 hours. The cosmetic composition containing pseudo-ceramide and ceramide NP were each added to fresh medium at 20 μg / mL and cultured for 24 hours. After lysing the cultured cells of each experimental group, the proteins were collected, and the amount of FLG produced was measured by performing an ELISA assay using a Human Filaggrin (FLG) ELISA kit (Cusa Biotechnology, USA) according to the manufacturer's manual. The measurement results are shown in Figure 5.

[0131] As shown in Figure 5, it was confirmed that the FLG production amount of the similar ceramide cosmetic composition increased by about 5.65%, and it was confirmed that it was statistically significantly higher than that of commercial ceramide NP.

[0132]

[0133] Test Example 3: Confirmation of the inhibitory effect of pseudo-ceramide complex on Prostaglandin E2 (PGE2) production

[0134] The anti-inflammatory effect of the pseudo-ceramide complex of Example 1 above was confirmed as follows.

[0135] RAW 264.7 Cell 1.0 X 10 5 Cells / ml were inoculated into a 6-well plate using DMEM (Dulbecco's Modified Eagle Medium) and cultured for 24 hours. The cosmetic composition containing pseudo-ceramide and ceramide NP were each added to fresh medium at 30 μg / mL, and lipopolysaccharides (LPS, Sigma-Aldrich) were added at 1 μg / mL, followed by incubation for 24 hours. The PGE2 content in the cell culture medium of each experimental group was measured by performing an ELISA assay according to the manufacturer's manual using a PGE2 Parameter assay kit (R&D System, USA). The measurement results are shown in Figure 6.

[0136] As shown in Figure 6, it was confirmed that the amount of PGE2 produced by the pseudo-ceramide cosmetic composition decreased by about 17.3%, and it was confirmed that the anti-inflammatory effect was superior to that of commercial ceramide NP.

[0137]

[0138] Test Example 4: Primary human skin irritation test of a cream formulation using a cosmetic composition containing pseudo-ceramide

[0139] The primary skin irritation caused by the cosmetic composition containing the pseudo-ceramide complex of Example 2 above was confirmed.

[0140] The study was conducted on 31 healthy men and women who had no acute or chronic renal diseases, including skin diseases, and whose body weight was not excessively abnormal.

[0141] After removing impurities from the subject's back (excluding the spine) using a test tissue (Kimtech), a patch inoculated with the product was applied. The patch was closed for 24 hours; after removing the patch, the test site was marked, and a first evaluation was conducted 30 minutes after removal, followed by a second evaluation 24 hours after removal. The average skin reactivity was calculated by combining the two evaluations of the product, and a final evaluation was conducted based on this result. The evaluation procedure was as shown in Table 3 below. All evaluations were performed under constant temperature and humidity conditions (22±2℃, 50±10% RH) free from air movement and direct sunlight, after the subject had rested for at least 30 minutes.

[0142] Test Phase Before Patch Application (0 Hours) 30 Minutes After Patch Removal (24 Hours) 24 Hours After Patch Removal (48 Hours) Subject Selection and Basic Investigation O Test Substance Closure Patch O 1st Judgment Evaluation O 2nd Judgment Evaluation O

[0143] The patch application site was selected as the back (excluding the spine). After applying 20 µl of the product to the patch, the patch was attached to the back (excluding the spine). For the negative control group, 20 µl of distilled water was applied, and the blank space within the patch where nothing was applied was selected as the Control. Skin reactivity was calculated according to the formula in Table 4 below.

[0144]

[0145] The final judgment criteria for the product were determined based on the average skin reactivity (Mean score) value, which is calculated by dividing the sum of each skin reactivity by 2, and the final judgment was made according to the criteria shown in Table 5 below.

[0146] Grade Mean Score No irritation 0.0 Low irritation 0.1 ~ 0.9 Mild irritation 1.0 ~ 2.9 Moderate irritation 3.0 ~ 4.9 Strong irritation 5.0 ~

[0147] The judgment results were as shown in the following Table 6.

[0148] Sample Responder Count Skin Reactivity Average Skin Reactivity Assessment 30 min 24 hr Example 200000 Comparative Example 100000 Blank (Control) 00000 Distilled Water (Negative Control) 00000

[0149] As a result of the test, the average skin reactivity of Example 2, a cream formulation containing a cosmetic composition with similar ceramide, and Comparative Example 1, a cream formulation containing ceramide NP, was calculated to be 0, and it was determined to be non-irritating according to the final judgment criteria.

[0150]

[0151] Test Example 5: Measurement of human skin moisture content of a cream formulation applied with a cosmetic composition containing pseudo-ceramide

[0152] The skin moisture content was confirmed by a cosmetic composition containing the pseudo-ceramide complex of Example 2 above.

[0153] The study was conducted on 11 healthy adult women aged 20 to 55 who had no acute or chronic renal diseases, including skin diseases, and whose body weight was not excessively abnormal.

[0154] The test areas were both sides of the face, and the product was randomly assigned to the corresponding areas for the subjects. Before using the product (Week 0), the subjects received the test formulation along with instructions for use of the cream formulation containing the cosmetic compositions of Example 2 and Comparative Example 1, and used it twice a day (morning and evening) for one week. The evaluation points were before using the test formulation (Week 0), immediately after using the test formulation, and at Week 1. At each evaluation point, skin moisture content was measured after the subjects had rested for at least 20 minutes under constant temperature and humidity conditions (22±2℃, 50±10% Rh) free from air movement and direct sunlight. Skin moisture content to evaluate skin moisturizing ability was measured using an Epsilon E100 (Biox, England). The measurement results are shown in Table 7 below.

[0155] Sample Time Mean ± Standard Deviation Change Rate (%) p-value Example 2 Before 23.05 ± 4.953 -- After 36.39 ± 8.786 ▲ 60.66 < 0.001 *** Week 144.06 ± 6.388▲96.82<0.001 *** Comparative Example 1 Before 23.97 ± 5.303 -- After 31.77 ± 6.669 ▲ 35.44 0.002 ** Week 137.45 ± 5.408▲61.90<0.001 ***

[0156] -Before vs. After, Week 1, Paired t test, Significance: ** p<0.01, *** p<0.001. Skin moisture images were measured and compared before and after using the test formulation (week 0), immediately after using the test formulation, and at week 1 after use. As a result, Comparative Example 1, a cream formulation containing Ceramide NP, showed a statistically significant improvement, increasing by 35.44% immediately after using the test formulation and by 61.90% at week 1 after use compared to before using the test formulation (week 0) (p<0.001). Example 2, a cream formulation containing a cosmetic composition containing pseudo-ceramide, showed a statistically significant improvement, increasing by 60.66% immediately after using the test formulation and by 96.82% at week 1 after use compared to before using the test formulation (week 0) (p<0.001).

[0157]

[0158] Test Example 6: Measurement of Transepidermal Water Loss (TEWL) of a Cream Formulation Applying a Cosmetic Composition Containing Ceramide-like Material

[0159] The study was conducted on 11 healthy adult women aged 20 to 55 who had no acute or chronic renal diseases, including skin diseases, and whose body weight was not excessively abnormal.

[0160] The test sites were both sides of the face, and the test formulation was randomly assigned to the corresponding sites and provided to the subjects. Before using the test formulation (Week 0), the subjects received the test formulation along with instructions for use of the cream formulation containing the cosmetic compositions of Example 2 and Comparative Example 1, and used it twice a day (morning and evening) for one week. The evaluation points were before using the test formulation (Week 0), immediately after using the test formulation, and Week 1. At each evaluation point, transepidermal water loss was measured after the subjects had rested for at least 20 minutes under constant temperature and humidity conditions (22±2℃, 50±10% Rh) free from air movement and direct sunlight. Transepidermal water loss (TEWL) for evaluating skin barrier function improvement was measured using a Vapometer (Delfin, Finland). The measurement results are shown in Table 8 below.

[0161] Sample Time Mean ± Standard Deviation Change Rate (%) p-value Example 2 Before 15.93 ± 4.141 -- After 13.39 ± 3.276 ▼ 15.6 10.001 ** Week 111.71 ± 2.560▼25.58<0.001 *** Comparative Example 1 Before 15.79 ± 4.348 -- After 13.80 ± 3.184 ▼ 11.500.001 *** Week 112.57 ± 2.681▼19.09<0.001 ***

[0162] -Before vs. After, Week 1, Paired t test, Significance: ** p<0.01, ***p<0.001. Transepidermal water loss was measured before use of the test formulation (week 0), immediately after use of the test formulation, and at week 1 after use, and compared before and after. As a result, Comparative Example 1, a cream formulation containing Ceramide NP, showed a statistically significant improvement, decreasing by 11.50% immediately after use of the test formulation and by 19.09% at week 1 after use compared to before use of the test formulation (week 0) (p<0.001). Example 2, a cream formulation containing a cosmetic composition containing pseudo-ceramide, showed a statistically significant improvement, decreasing by 15.61% immediately after use of the test formulation and by 25.58% at week 1 after use compared to before use of the test formulation (week 0) (p<0.001).

[0163] As a result, as confirmed from the results of the above test examples, the cosmetic composition containing the ceramide-like substance of the present invention has a higher rate of change in skin moisture content (%) and a higher rate of change in transepidermal water loss (%) than the commercially available ceramide, ceramide NP, and is more effective in improving skin moisture content and skin barrier function.

[0164]

[0165] Test Example 7: Stability test of pseudo-ceramide complex

[0166] The pseudo-ceramide complex was stored for 4 weeks under conditions of -20℃, 4℃, 25℃, 45℃, and sunlight, and the changes in the content of each pseudo-ceramide component were checked at weekly intervals using gas chromatography.

[0167] Before proceeding with gas chromatography analysis, 1 μL of a pseudo-ceramide complex was collected and diluted 20 to 50 times with chloroform to prepare a sample for analysis.

[0168] The pseudo-ceramide complex was analyzed using gas chromatography (7890A, Agilent Technologies, Santa Clara, CA, USA) equipped with an HP-5 capillary column (30 m length x 0.320 mm diameter, 0.25 μm film thickness, Hewlett Packard, USA). The oven temperature was started at 150°C, maintained for 1 minute, and then raised to 350°C at a rate of 10°C / min (maintenance time 3 minutes). A flame ionization detector was used as the detector, with the detector temperature set to 270°C and the injector temperature to 350°C. Helium was used as the carrier gas. The analysis results are shown in Table 9 below.

[0169] Room Temperature (25℃) Ingredients Day 0 Day 7 Day 14 Day 21 Day 28 Oleic / Oleic amide ester 35.8 4 35.7 4 35.4 6 35.6 5 35.5 6 Palmitic / Oleic amide ester 19.2 3 19.2 3 19.4 2 19.2 3 19.2 3 Refrigerated (4℃) Ingredients Day 0 Day 7 Day 14 Day 21 Day 28 Oleic / Oleic amide ester 35.8 4 35.8 4 35.5 6 36.0 2 35.6 5 Palmitic / Oleic amide ester 19.2 3 19.1 4 19.1 4 18.7 6 19.0 4 Frozen (-20℃) Ingredients Day 0 Day 7 Day 14 Day 21 Day 28 Oleic / Oleic amide ester 35.8 4 35.5 6 35.6 5 35.6 5 35.5 6 Palmitic / Oleic amide ester 19.2 3 19.1 4 18.9 5 18.9 5 18.9 5 High temperature (45℃) component Day 0 Day 7 Day 14 Day 21 Day 28 Oleic / Oleic amide ester 35.8 4 35.6 5 35.4 6 35.4 6 35.3 7 Palmitic / Oleic amide ester 19.2 3 19.2 3 19.1 4 19.1 4 18.8 6 Sunlight component Day 0 Day 7 Day 14 Day 21 Day 28 Oleic / Oleic amide ester 35.8 4 35.5 6 35.3 7 35.3 7 35.2 7 Palmitic / Oleic amide ester19.2319.0418.9519.0418.58

[0170] (unit : %)

[0171] As a result of the stability test of the component content of the pseudo-ceramide complex, as shown in Table 9, the content of the two components, Oleic / Oleic amide ester and Palmitic / Oleic amide ester, on day 0 was 35.84% and 19.23%, respectively. When the content was checked at weekly intervals under conditions of -20℃, 4℃, 25℃, 45℃, and sunlight, it was confirmed that the stability of the pseudo-ceramide complex was excellent, with 35.27% and 18.58% at week 4.

[0172]

[0173] Test Example 8: Formulation stability test

[0174] The change in pH of a cream formulation containing a ceramide-like complex and a cream formulation containing commercial ceramide NP was checked under -20℃, 4℃, 25℃, 45℃, and sunlight conditions.

[0175] According to the formulation of Comparative Example 1 in Table 2, a cream formulation serving as the comparative group was prepared by varying only the content of ceramide NP to 0.5 wt%, 1 wt%, and 3 wt%. Similarly, according to the formulation of Example 2 in Table 2, a cream formulation serving as the control group was prepared by varying only the content of the pseudo-ceramide complex to 0.5 wt%, 1 wt%, and 3 wt%. The comparative and experimental groups were stored for 4 weeks under conditions of -20°C, 4°C, 25°C, 45°C, and sunlight, and pH changes were measured at weekly intervals (to measure pH fluctuations caused by impurities, etc., detached from the pseudo-ceramide). pH measurements were taken at weekly intervals for 4 weeks for the comparative and experimental groups under storage at different temperatures and sunlight exposure. Accuracy was ensured by pH correction using a pH standard solution prior to measurement, and the temperature was maintained at 25 ± 1°C during measurement. The results are shown in Table 10 below.

[0176] Comparative Group (Ceramide NP applied cream formulation) pH Day 0 Day 7 Day 14 Day 21 Day 28 Comparative Example 1 - 20℃ 6.4 4 6.4 7 6.5 26.6 56.7 24℃ 6.4 4 6.4 7 6.5 16.6 96.8 325℃ 6.4 4 6.4 6 6.4 6 6.5 06.4 745℃ 6.4 4 6.4 96.5 96.8 16.97 Sunlight 6.4 4 6.5 16.6 96.8 47.04 Comparative Example 2-20℃ 6.50 6.54 6.61 6.77 6.884℃ 6.50 6.49 6.60 6.87 7.0925℃ 6.50 6.47 6.53 6.53 6.5145℃ 6.50 6.50 6.68 6.91 7.17 Solar light 6.50 6.60 6.77 6.95 7.22 Comparative Example 3-20℃ 6.63 6.61 6.89 7.02 7.034℃ 6.63 6.84 6.91 7.11 7.3925℃ 6.63 6.66 6.65 6.60 6.6545℃ 6.63 6.78 6.88 7.27 7.40 Solar light 6.63 6.68 6.93 7.23 7.49 Control group (similar Ceramide-applied cream formulation) pH Day 0 Day 7 Day 14 Day 21 Day 28 Example 2 - 20℃ 6.58 6.60 6.60 6.65 6.654℃ 6.58 6.60 6.60 6.62 6.6725℃ 6.58 6.56 6.62 6.62 6.6145℃ 6.58 6.57 6.65 6.67 6.70 Sunlight 6.58 6.59 6.68 6.74 6.77 Example 3-20℃6.546.576.556.616.604℃6.546.486.546.606.6325℃6.546.556.586.556.6045℃6.546.536.556.606.68 Solar light 6.546.596.666.726.70 Example 4-20℃6.476.516.496.546.514℃6.476.536.486.516.4625℃6.476.506.526.496.5845℃6.476.446.406.476.59 Solar light 6.476.536.596.656.63

[0177] In the control group formulations containing 0.5%, 1%, and 3% ceramide NP, the initial pH increased from 6.44, 6.50, and 6.63 to approximately 0.2 to 1.0 after 4 weeks under all conditions except 25℃.

[0178] There was almost no change in pH at 25℃. In particular, pH increased most significantly at 45℃ and under sunlight conditions.

[0179] In the control formulations with 0.5%, 1%, and 3% ceramide-like compounds, the initial pH was 6.58, 6.54, and 6.47, and after 4 weeks, it increased slightly by about 0.05 to 0.2 in all conditions.

[0180] In conclusion, the control group formulation with ceramide analogue had superior pH stability compared to the comparison group formulation with ceramide NP.

[0181] Fluctuations in pH values ​​indicate that fatty acids are derived from pseudo-ceramide, and the range of fluctuation in the example was much lower than that of Comparative Example 1. In particular, the difference was pronounced at low and high temperatures, and since cosmetics containing pseudo-ceramide are often stored in cosmetic refrigerators, formulation stability at low temperatures is also of great significance.

[0182] The present invention can be widely utilized in the fields of cosmetics, beauty, and the upcycling industry.

Claims

1. A step of adding vegetable oil, amino alcohol, and lipase to an organic solvent and reacting them; and The method sequentially includes the step of adding additional unsaturated fatty acids and lipase to the above reactants and reacting them again, wherein A method for manufacturing a pseudo-ceramide complex characterized in that the vegetable oil is yuzu seed oil.

2. A method for preparing a pseudo-ceramide complex according to claim 1, wherein the yuzu seed oil contains at least 35% oleic acid and at least 20% palmitic acid.

3. A method for preparing a pseudo-ceramide complex according to claim 1, wherein the yuzu seed oil has a saponification value of 180 to 210, an iodine value of 90 to 110, and an acid value of 0.3 to 0.

6.

4. A method for preparing a pseudo-ceramide complex according to claim 1, characterized in that the additional unsaturated fatty acid is oleic acid.

5. A method for producing a pseudo-ceramide according to claim 1, characterized in that the amino alcohol is 3-amino-1-propanol.

6. A method for preparing a pseudo-ceramide complex according to claim 1, characterized in that the organic solvent is hexane.

7. A method for preparing a pseudo-ceramide complex according to claim 1, characterized in that the lipase is a CAL-B enzyme or a variant thereof.

8. A ceramide-like complex derived from yuzu seed oil, prepared according to the method of claim 1.

9. A citron seed oil-derived pseudo-ceramide complex according to Claim 8, characterized by being represented by the following chemical formula 1. [Chemical Formula 1] Here, R in the above chemical formula 1 , or R 2 Each is an acyl residue derived from the fatty acid or oleic acid of citron seed oil, independently.

10. A citron seed oil-derived pseudo-ceramide complex according to claim 8, characterized by comprising any one or more compounds represented by the following chemical formulas 2 to 6. [Chemical Formula 2] [Chemical Formula 3] [Chemical Formula 4] [Chemical Formula 5] [Chemical Formula 6] 11. A citron seed oil-derived pseudo-ceramide complex according to claim 10, characterized in that it contains 35 to 40 weight percent of the compound of formula 2 based on the total weight of the composition.

12. A citron seed oil-derived pseudo-ceramide complex according to claim 10, characterized in that it contains 35 to 40 weight percent of the compound of formula 2 based on the total weight of the composition.

13. A cosmetic composition containing 0.001 to 10% by weight of the citron seed oil-derived ceramide complex of claim 8 or claim 9 based on the total weight of the cosmetic composition.

14. In claim 13, caprylic / capric triglyceride 6-10 wt%, coco-caprylate / caprate 3-7 wt%, 2,3-butanediol 3-6 wt%, glycerin 1-5 wt%, pentaerythrityl distearate 1-3 wt%, cetearyl glucoside 0.5-2.5 wt%, cetearyl alcohol 0.5-2.5 wt%, shea butter 0.5-2.5 wt%, vinyl dimethicone 0.5-2.5 wt%, 1,2-hexanediol 1-2 wt%, stearyl alcohol 0.1-0.5 wt%, stearic acid 0.1-0.3 wt%, tromethamine 0.1-0.4 wt%, sodium stearoyl glutamate A cosmetic composition characterized by a cream formulation further comprising 0.1~0.3 wt%, 0.2~0.4 wt% carbomer, 0.01~0.03 wt% disodium EDTA, 0.02~0.04 wt% fragrance, and 65~75 wt% purified water.

15. A cosmetic composition according to claim 13, characterized in that the cosmetic composition is for strengthening the skin barrier, for alleviating skin inflammation, or for moisturizing the skin.

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