JP2020045308A - Novel ceramide and skin humectant containing the same and method for producing ceramide composition - Google Patents

Abstract

To provide a production method that produces a novel ceramide having a similar structure to that of ceramide on human skin by a safe and convenient method without using harmful extraction solvent from natural glucosylceramide.SOLUTION: The present invention relates to a ceramide represented by formula (I) (R, R, Rand Rindependently represent H, a methyl group, an ethyl group or an acetyl group; wave lines denote that adjacent double bonds are E or Z arrangements, n is an integer of 1-11).SELECTED DRAWING: Figure 1

Description

  The present invention relates to a novel ceramide, a naturally derived ceramide composition containing the same, their use as a skin moisturizer, and a method for producing a ceramide composition.

  Since the skin is located at the boundary between the outside world and the living body, it has a defense mechanism against constant external stimuli such as ultraviolet rays, oxidative stress, dryness, chemical substances and microorganisms. Ceramide, one of the sphingolipids, is present in the stratum corneum of the skin and accounts for about 50% of the intercellular lipids in the stratum corneum. Sphingolipid is a generic term for complex lipids containing a sphingosine skeleton. Glucosylceramide is a kind of sphingolipid in which glucose is bound to ceramide. Ceramide synthesized in skin epidermal cells is once stored in the form of glucosylceramide or sphingomyelin, then excreted out of the cell and subjected to the action of β-glucocerebrosidase (β-glucosidase) and sphingomyelinase. It becomes ceramide again and exhibits the function of a permeation barrier in the stratum corneum of the epidermis.

  Ceramide has a structure in which fatty acids are bonded to sphingosine, which is a long-chain base. Keratinocytes in late differentiation (granular layer) synthesize various ceramides, and the diversity of these molecules is due to the hydroxylation (non-hydroxy, 2-hydroxy and terminal ω-hydroxy) of fatty acids and sphingosine bases constituting ceramide. Due to diversity. In addition to the diversity of hydroxylation, there is also diversity in the chain length of fatty acids and sphingosine bases. These ceramides are found not only in the skin but also in blood, brain, spinal cord and nervous tissue in vivo, and in plants, they are contained in wheat, rice, soybean, millet, spinach, mushrooms and the like.

  Methods for extracting ceramide from plant materials include, for example, a method of extracting seeds such as yuzu, grapes and cherries, and plant materials such as wheat, rice and soybean with an alcoholic solvent (eg, ethanol) (see Patent Document 1). There are known methods for hydrolyzing plant materials such as wheat, soybean, corn, and rice bran with an alkaline ethanol solvent, followed by extraction with a mixed solvent of hexane, acetone and water (for example, see Patent Documents 2 and 3). Vegetable ceramides are plant-derived and therefore have a characteristic that they are gentle to the body and do not easily irritate when formulated into cosmetics or supplements.However, because of plant-derived components, ceramides on human skin have a subtle structure. different. It is not clear whether plant ceramides extracted from plants such as rice, corn and soybean have the same skin barrier function as ceramide present in the skin stratum corneum.

A method for producing ceramide from glucosylceramide by a deglucosidation reaction has already been reported (see Non-Patent Documents 1 to 3). Non-Patent Document 1 discloses a method of preparing ceramide by acid hydrolysis under mild conditions by performing periodate oxidation and NaBH ₄ reduction from galacto or glucocerebroside to decompose a sugar moiety, and under heating conditions. A method for N-acylation of sphingosine prepared from the acid hydrolyzate in Example 1 to reconstruct ceramide is described. Non-Patent Document 2 describes a method for preparing ceramide under mildly acidic conditions after periodate oxidation from brain-derived cerebroside in a chloroform / ethanol solution and reduction of NaBH ₄ . Non-Patent Document 3 reports the results of decomposing cerebroside derived from sea anemone under the same conditions as described above, and analyzing the structure.

Japanese Patent No. 4,108,069 JP-A-2002-030993 JP 2006-232699A

Iga, S .; et al. , Preparation of Ceramide and Sphingosine by Chemical and Biochemical Methods. Romanian Biotechnological Letters, Vol. 16, No. 6, pp. 6841-6846 Carter, HE. et al. , Biochemistry of the sphingolipids: XII. Conversion of cerebrosides to ceramides and sphingosine; structure of Gaucher cerebroside. J. Lipid Research 1961, Vol. 2, No. 3, pp. 228-234. Karlsson KA, Leffler H, Samuelsson BE. Characterization of cerebroside (monoglycosylceramide) from the sea anemone, Medium medium series. Identification of the major long-chain base as an unusual dienic base with a methyl branch at a double bond. Biochim Biophys Acta. 1979, Jul 27; 574 (1): 79-93.

  However, the methods described in the above Non-Patent Documents 1 to 3 use a highly toxic solvent such as chloroform, do not describe the reaction yield, and do not optimize the reaction conditions. According to such a conventional production method, there is a problem that the yield of ceramide is small, and much labor and cost are required for extraction and purification. An object of the present invention is to obtain a novel ceramide having a structure similar to that of ceramide present in human skin from naturally occurring glucosylceramide by a safe and simple method without preferably using a harmful extraction solvent. And one of them.

  The present invention has been made to solve such a problem.

（式中、Ｒ^１、Ｒ^２、Ｒ^３およびＲ^４は、互いに独立して水素原子、メチル基、エチル基またはアセチル基を表し、波線は、隣接する二重結合がＥまたはＺの配置であることを表し、ｎは、１〜１１のいずれかの整数を表す。）で示されるセラミドである。 That is, one embodiment of the present invention provides the following formula (I):

(Wherein R ¹ , R ² , R ³ and R ⁴ independently represent a hydrogen atom, a methyl group, an ethyl group or an acetyl group, and the wavy line indicates that the adjacent double bond is in the E or Z configuration. And n represents an integer of any of 1 to 11).

  In another embodiment, the present invention relates to a composition comprising a plurality of ceramides represented by the above formula (I), wherein the chain length of the fatty acid moiety and the arrangement of E or Z in the ceramide mixture are varied by changing the number of n. A rice-derived ceramide composition is provided, wherein the proportion is related to their abundance in the rice ceramide.

  According to a different aspect of the present invention, there is provided a skin moisturizer comprising the ceramide or the ceramide composition as an active ingredient.

（式中、波線は、隣接する二重結合がＥまたはＺの配置であることを表し、ｎは、１〜１１のいずれかの整数を表す。）に示すセラミドを得る工程と、
を含む、セラミド組成物の製造方法を提供する。 In a further different aspect, there is provided a method for producing the ceramide composition,
Prepare glucosylceramide extracted from rice,
Oxidizing the glucosylceramide to an aldehyde intermediate by reacting the glucosylceramide and sodium periodate in an aqueous medium containing an organic solvent miscible with water,
In the water-miscible organic solvent, reducing the aldehyde intermediate by reacting the aldehyde intermediate and sodium borohydride,
An acid and water are added to the reduction reaction product of the aldehyde intermediate to carry out a hydrolysis reaction, and the following formula (II):

(In the formula, a wavy line indicates that an adjacent double bond has an E or Z configuration, and n represents an integer of any of 1 to 11).
A method for producing a ceramide composition is provided.

  ADVANTAGE OF THE INVENTION According to the manufacturing method of this invention, ceramide excellent in safety can be manufactured from a plant raw material by a simple method, preferably without using harmful chemicals. Moreover, since the novel ceramide of the present invention is structurally similar to ceramide present in the stratum corneum of human skin, it is expected to have excellent skin moisturizing properties.

FIG. 1 is a scheme of a method for deglycosylating glycosylceramide by the production method of the present invention. In the title of the figure, “1” is compound 1, “2” is compound 2, and “4” is compound 4. FIG. 2 shows the result of HPLC analysis of compound 4 obtained in Example 7.

  Hereinafter, the novel ceramide according to the present invention and a ceramide composition containing the same will be described first, followed by a description of its production method and its use as a skin moisturizer.

(Ceramide and ceramide composition)
In the present specification, ceramide is a kind of sphingolipid, and is sphingosine and a fatty acid in which an amide bond is formed. Also, glucosyl ceramide is a compound in which glucose is bound to ceramide. One embodiment of the present invention includes a ceramide represented by the following formula (I).

（式中、Ｒ^１、Ｒ^２、Ｒ^３およびＲ^４は、互いに独立して水素原子、メチル基、エチル基またはアセチル基を表し、波線は、隣接する二重結合がＥまたはＺの配置であることを表し、ｎは、１〜１１のいずれかの整数を表す。）で示されるセラミドである。すなわち、式（Ｉ）で表されるセラミドにおいてスフィンゴシン由来の長鎖塩基は、２つの二重結合に基づく幾何異性体を含んでおり、一方（４位）の二重結合はＥ（ｅｎｔｇｅｇｅｎ）であり、他方（８位）の二重結合はＥおよびＺ（ｚｕｓａｍｍｅｎ）の何れかとして存在してもよい。したがって、このような異性体およびそれらの混合物のすべては、本発明の範囲内に含まれる。

(Wherein R ¹ , R ² , R ³ and R ⁴ independently represent a hydrogen atom, a methyl group, an ethyl group or an acetyl group, and the wavy line indicates that the adjacent double bond is in the E or Z configuration. And n represents an integer of any of 1 to 11). That is, the sphingosine-derived long-chain base in the ceramide represented by the formula (I) contains a geometric isomer based on two double bonds, while the double bond at position (4) is E (entgegen). Yes, the double bond at the other (position 8) may be present as either E or Z (zusammen). Accordingly, all such isomers and mixtures thereof are included within the scope of the present invention.

In formula (I), ceramide in which R ¹ , R ² , R ³ and R ⁴ are all hydrogen atoms can be produced by deglucosylating glucosylceramide extracted from rice by a method described later. . A derivative in which any or all of R ¹ , R ² , R ³ and R ⁴ are substituted with an alkyl group such as a methyl group or an ethyl group can be obtained by converting a ceramide derived from rice into an alkylating agent in the presence of a base. To act. Examples of the base used include inorganic bases such as sodium hydroxide, sodium carbonate and potassium carbonate, alkali metal alkoxides such as sodium methoxide, sodium ethoxide, sodium isopropoxide and potassium tert-butoxide, sodium hydride, potassium hydride and the like. Of these, sodium carbonate, sodium methoxide, sodium ethoxide, potassium tert-butoxide, and sodium hydride are preferable from the viewpoint of yield and economy.

  Examples of the alkylating agent to be used include alkyl halides such as methyl iodide, ethyl iodide, methyl bromide, ethyl bromide, benzyl bromide, methyl chloride, ethyl chloride and benzyl chloride, and sulfate esters such as dimethyl sulfate and diethyl sulfate. And sulfonic acid esters such as methyl methanesulfonate, ethyl methanesulfonate, methyl p-toluenesulfonate, ethyl p-toluenesulfonate, and methyl p-trifluoromethanesulfonate. Preferred are methyl iodide, ethyl iodide, methyl bromide, ethyl bromide, benzyl bromide, benzyl chloride, methyl methanesulfonate, ethyl methanesulfonate, methyl p-toluenesulfonate, dimethyl sulfate, and diethyl sulfate. Among them, methyl iodide, ethyl iodide, dimethyl sulfate and diethyl sulfate are particularly preferred.

A derivative in which any or all of R ¹ , R ² , R ³ and R ⁴ are acetylated can be obtained by allowing an acetylating agent such as acetic anhydride or acetyl chloride to act.

  The ceramide represented by the above formula (I) contains a plurality of ceramides depending on the variation of the number n and the difference in the arrangement of E or Z. Accordingly, another embodiment of the present invention is a composition comprising a plurality of ceramides represented by the above formula (I), wherein the number of n in such a ceramide mixture varies, and the chain length of the fatty acid moiety, and Ceramide compositions in which the proportion of the E or Z configuration correlates with their abundance in plant ceramides, especially rice ceramides. Ceramides extracted from plant raw materials including rice are derived from natural materials, and are therefore desired to be used in health care products from the viewpoint of safety and cost. However, it has been pointed out that ceramides derived from these plants have slightly different structures from ceramides present in the stratum corneum of the skin. The ceramide composition of the present embodiment has a structure similar to human skin stratum corneum ceramide and, because it is a ceramide derived from a natural material, has not only an effect such as moisturizing property and stability but also safety. It is excellent. Such a ceramide composition can be produced by the following method.

(Method for producing ceramide composition)
Glucosylceramide used in the present embodiment is typically glucosylceramide derived from rice, but may be glucosylceramide derived from other plants and the like, for example, grape, cherry, yuzu, olive, perilla, shikuwasa , Prunes, wheat, wheat germ, millet, soybeans, sesame, peanuts, corn, spinach, konjac potato, pineapple, sugar beet, sugar beet, Unshu mandarin orange, peach, mushrooms (such as Maitake mushrooms, Tamagitake mushrooms), milk and other extracts Is mentioned. Methods for extracting glucosylceramide from plant raw materials are known. For example, extraction is performed by the method described in JP-A-2001-09983, or commercially available rice-derived glucosylceramide (eg, Nagara Science Co., Ltd.) Can be used.

  Examples of the organic solvent for obtaining the extract include water, alcohols (eg, anhydrous ethanol, ethanol, propylene glycol, 1,3-butylene glycol, etc.), ketones such as hexane and acetone, diethyl ether, dioxane, and ethyl acetate. Organic solvents such as esters and the like can be used alone or in any combination of two or more kinds of liquid mixtures, and can also be used in a state where each solvent extract is combined. Preferably, ethanol is used. The amount of the solvent used is 1 to 100 parts by weight, preferably 10 to 30 parts by weight, per 1 part by weight of the plant material for extraction.

  Further, an alkaline organic solvent obtained by adding potassium hydroxide, sodium hydroxide, or the like to these alcohol solvents to make them alkaline may be used. The preferred alkali concentration is 0.01 to 1N, and particularly preferably 0.1 to 0.4N.

  The extraction method is not particularly limited, but usually, it may be within the range of the boiling point of the solvent at normal temperature and normal pressure, and filtration may be performed after extraction. What is necessary is just to arbitrarily select a normal means generally applied in the extraction processing.

  The glucosylceramide prepared by these methods is deglucosylated in the three-step process shown in FIG. 1 to prepare a ceramide composition containing a novel ceramide. In FIG. 1, compound 1 is glucosylceramide derived from rice prepared by the above method. According to the findings of the present inventors, rice glucosylceramide is composed of a mixture of α-hydroxy acids having different chain lengths, and contains a plurality of saturated fatty acids having 15 to 25 carbon atoms (n = 1 to 11). . Above all, fatty acids having 18 to 24 carbon atoms (n = 4 to 10) are contained in a large amount, and fatty acids having 18 and 20 carbon atoms are the largest.

  The glucosylceramide derived from rice can be oxidized to an aldehyde intermediate represented by compound 2 in FIG. 1 by reacting it with sodium periodate in an aqueous medium containing an organic solvent miscible with water. The solvent used in this oxidation reaction is not particularly limited as long as it is an aqueous medium containing an organic solvent miscible with water, but typically, a mixed solvent of water and ethanol is used. The mixing ratio is not particularly limited, but an aqueous solution of 30% to 90% by volume of ethanol can be used, an aqueous solution of 50% to 70% by volume of ethanol is preferable, and an aqueous solution of about 60% by volume of ethanol is most preferable. By this oxidation reaction, the carbon-carbon bond having a 1,2-diol (glycol) structure in the glucose molecule is cleaved to obtain compound 2 in FIG. Although the reaction time and reaction temperature are not particularly limited, the reaction may be performed at room temperature (about 16 ° C. to 27 ° C.) for 1 to 24 hours. By using sodium periodate, a reaction under mild conditions at a pH in a neutral region is possible. After the reaction, it is preferable to deactivate the remaining sodium periodate by adding glycerin or sodium L-ascorbate. In particular, the sodium periodate excessively added by the addition of sodium L-ascorbate is completely deactivated. Can be alive.

Subsequently, the aldehyde intermediate represented by the compound 2 is reacted with sodium borohydride in the water-miscible organic solvent to reduce the aldehyde intermediate. The reduction reaction product is subjected to a hydrolysis reaction by adding an acid and water to give a compound 4 represented by the following formula (II):
The following formula (II):

（式中、波線は、隣接する二重結合がＥまたはＺの配置であることを表し、ｎは、１〜１１のいずれかの整数を表す。）を得ることができる。本実施形態では、温和な条件を用いることができるため、分解されたグルコース以外の部分は、天然物であるコメ由来のセラミドの化学構造が維持され、特に、脂肪酸の鎖長が長いセラミドは保湿性に有効と考えられる。

(In the formula, a wavy line indicates that the adjacent double bond has an E or Z configuration, and n represents an integer of any of 1 to 11.) In this embodiment, since mild conditions can be used, the parts other than degraded glucose maintain the chemical structure of ceramide derived from rice, which is a natural product, and particularly, ceramide having a long fatty acid chain length is moisturized. It is considered effective for sex.

(Skin moisturizer)
In another embodiment of the present invention, there is provided a skin moisturizer containing the ceramide or the ceramide composition obtained by the above-mentioned production method as an active ingredient. The ceramide obtained in Example 3 described later becomes almost a single spot and peak by TLC and liquid chromatography. Further, the skin moisturizer of the present embodiment is a cosmetic composition such as an external preparation composition, a food composition such as a health food, or an intermediate material thereof, in the form of powder, liquid, granule, tablet, paste, or emulsion. , Creams, gels, capsules, aerosols, and the like, and an optimal shape for constituting a final product can be arbitrarily selected.

When preparing a food composition, the form can be appropriately selected, and a beverage is also included.
In addition to general foods, treatment, prevention or improvement by promoting the production of ceramide, such as improvement of keratinization of the epidermis, improvement and maintenance of skin moisturizing function and skin barrier function, imparting of firmness or firmness of hair, and improving feeling of hair. Foods and drinks that indicate the concept of treating, preventing or ameliorating a possible disease or condition, such as health foods, functional foods, foods for the sick, and foods for specified health use are also included. Health foods, functional foods, foods for the sick, and foods for specified health use are specifically formulated as various preparations such as fine granules, tablets, granules, powders, capsules, syrups, liquids, and liquid foods. And can be used for these formulations. The food composition in the form of a formulation is prepared by mixing the active ingredient with a carrier acceptable as a food, for example, a suitable excipient (eg, starch, processed starch, lactose, glucose, water, etc.), and then mixing the active ingredient with a conventional means. Can be manufactured. Furthermore, food compositions include soups, juices, milk drinks, tea drinks, coffee drinks, cocoa drinks, liquid food compositions such as jelly drinks, semi-solid food compositions such as pudding, yogurt, breads, udon Noodles, cookies, chocolate, candy, gum, senbei and other confectionery, sprinkles, butter, jams and other spreads. Food also includes feed.

  Food compositions include various food additives, for example, antioxidants, flavors, various esters, organic acids, organic acid salts, inorganic acids, inorganic acid salts, inorganic salts, pigments, emulsifiers, preservatives, seasonings Additives such as ingredients, sweeteners, sour agents, fruit juice extracts, vegetable extracts, nectar extracts, pH adjusters, quality stabilizers and the like may be used alone or in combination.

  Examples of the external preparation composition include lotions, emulsions, creams, ointments, lotions, oils, basic cosmetics such as packs, soaps, cleansing creams, cleansing lotions, skin cleansers such as facial cleansers, shampoos, rinses, and treatments. Cosmetics for hair washing, hair creams, hair sprays, hair tonics, hair gels, hair lotions, hair oils, hair essences, hair styling products such as hair water, hair wax, hair foam, etc., hair restoration / hair restoration, foundation, white powder, whitening Makeup cosmetics such as lipstick, blusher, eye shadow, eyeliner, mascara, eyebrows, eyelashes, finishing cosmetics such as nail polish, and cosmetics such as oral compositions such as perfumes, toothpastes, gargles, etc. Composition, topical medicinal preparations, ointments, cataplasms, bath preparations, medicated toothpaste, mouthwash, etc. Cavity compositions, medicated cosmetics, hair dyes, hair restorers, hair loss inhibitors, deodorants in hair solvents such as hair removers, quasi-drugs for external use such as hygiene products, sanitary cotton, wet tissue, Topical drugs and the like.

  The concentration of ceramide in the food composition and the external preparation composition according to the present embodiment is about 0.00001 to 100% by mass (hereinafter, expressed as%), preferably about 0.0005 to 50%, as solid content. If so, usability and good effects can be obtained.

  In addition, the skin moisturizing agent of this embodiment, the food composition containing this, and the external preparation composition can also be manufactured using together the components and additives normally used for these. For example, it can be produced by arbitrarily selecting and using the components and additives exemplified below, and the content in the preparation is not particularly limited, but is usually preferably 0.0001 to 50%.

  The external preparation composition comprises other components commonly used in cosmetics, quasi-drugs, pharmaceuticals, and the like, for example, powder components, liquid fats and oils, solid fats and oils, waxes, hydrocarbons, higher fatty acids, higher alcohols, esters, silicones, and anions. Surfactants, cationic surfactants, amphoteric surfactants, nonionic surfactants, humectants, water-soluble polymers, thickeners, coating agents, ultraviolet absorbers, sequestering agents, lower alcohols, polyhydric alcohols , Sugars, amino acids, organic amines, polymer emulsions, pH adjusters, skin nutritional supplements, vitamins, antioxidants, antioxidant aids, fragrances, water, etc., if necessary, and can be produced by a conventional method. it can.

  Other components that can be incorporated into the external preparation composition include, for example, preservatives (eg, ethyl paraben, butyl paraben), anti-inflammatory agents (eg, glycyrrhizic acid derivatives, glycyrrhetinic acid derivatives, salicylic acid derivatives, hinokitiol, zinc oxide, allantoin, etc.) ), Whitening agents (for example, ascorbic acid and derivatives thereof, placenta extract, Saxifraga extract, arbutin, etc.), various extracts (for example, oak, oak, sicon, peony, assembly, birch, sage, loquat, carrot, aloe) , Mallow, iris, grape, yokuinin, loofah, lily, saffron, senkyu, ginger, hypericum, ononis, garlic, capsicum, chimpanzee, corn, seaweed, etc., activator (for example, royal jelly, sensitizer, cholesterol derivative etc.) Blood circulation enhancer For example, nonylate valenylamide, nicotinic acid benzyl ester, nicotinic acid β-butoxyethyl ester, capsaicin, gingerone, cantharistinki, itactamol, tannic acid, α-borneol, tocopherol nicotinate, inositol hexanicotinate, cyclandate, cinnarizine , Trazoline, acetylcholine, verapamil, cepharanthin, γ-oryzanol, etc.), antiseborrheic agents (eg, sulfur, thiantol, etc.), anti-inflammatory agents (eg, tranexamic acid, thiotaurine, hypotaurine, etc.) and bactericides (eg, triclosan, Cetylpyridinium chloride, thymols, benzalkonium chloride, and the like).

  The subject to which the skin moisturizing agent of the present embodiment is administered is preferably a warm-blooded vertebrate, and more preferably a mammal. In the present specification, examples of the mammal include humans and non-human mammals such as monkeys, mice, rats, rabbits, dogs, cats, cows, horses, and pigs. Ceramide production enhancer, cholesterol production enhancer, intercellular lipid production enhancer, skin barrier function improver, epidermal keratinization improver, skin moisturizer, skin roughening preventive or improver of the present invention, hair firmness or firming agent Or a hair feel improving agent is suitable for administration to humans, primates such as monkeys, and particularly to humans.

  The skin moisturizing agent of the present embodiment is desired to improve the skin barrier function, prevent or treat keratinization deficiency of the epidermis, moisturize the skin, prevent or improve rough skin, impart hair firmness or firmness, or improve the feel of hair. It can be applied to those who do. The skin moisturizer is preferably applied under necessary conditions (preferably, low humidity and dry conditions). The skin moisturizer is preferably applied to the skin, scalp or hair.

  The dose of the active ingredient in the skin moisturizer of the present embodiment is appropriately determined depending on the condition of the individual, body weight, sex, age, activity of the material, administration or intake route, administration or intake schedule, formulation form or other factors. be able to. For example, based on the mass of the active ingredient, preferably 0.001 mg or more, more preferably 0.01 mg or more, preferably 1000 mg or less, more preferably 100 mg or less, more preferably 100 mg or less per adult (60 kg in weight) per day. 0.001 to 1000 mg, more preferably 0.01 to 100 mg. Further, the active ingredient may be ingested / administered once to several times a day, or at any time and interval.

  The content of the active ingredient in the skin moisturizing agent of the present embodiment can be appropriately determined so as to achieve the above-mentioned dose. For example, in the total amount of the skin moisturizer of the present embodiment, the content of the active ingredient is preferably 0.001% by mass or more, more preferably 0.01% by mass or more, and preferably 10% by mass or less, and more preferably 5% by mass. The following is more preferable, 0.001 to 10% by mass is preferable, and 0.01 to 5% by mass is more preferable.

[Example 1] Synthesis of compound 2-1
Suspended in distilled water (48 mL) and ethanol (70 mL) in hydrated glucosylceramide (64.0 g, solid content 10.9%). Sodium periodate (11.6 g) was added at 26-27 ° C over 23 minutes and stirred at room temperature for 13 hours. After glycerol (12.3 g) was added at 23 to 31 ° C. over 25 minutes, water (250 mL) and ethyl acetate (500 mL) were added, and the mixture was separated. The organic layer was washed with distilled water (250 mL), and the aqueous layers were combined and extracted with ethyl acetate (500 mL). The organic layers were combined, anhydrous sodium sulfate (150 g) was added, the mixture was stirred, filtered, and washed with ethyl acetate. Concentration at 50 ° C. under reduced pressure gave brown amorphous compound 2 (6.22 g).

Example 2 Synthesis of Compound 2-2
Suspended in distilled water (48 mL) and ethanol (70 mL) in hydrated glucosylceramide (64.0 g, solid content 10.9%). Sodium periodate (11.6 g) was added over 4 minutes at 24-30 ° C and stirred at room temperature for 18 hours. After adding sodium L-ascorbate (9.56 g) / distilled water (38 mL) at 20 to 30 ° C. over 20 minutes (colorless with iodine starch paper), the mixture was concentrated at 50 ° C. under reduced pressure to 180 g. (Brown and wax-like substance was precipitated) Water (100 mL) and ethyl acetate (100 mL) were added, and the mixture was separated. The aqueous layer was extracted with ethyl acetate (50 mL). The organic layers were combined, anhydrous sodium sulfate (7.5 g) was added, the mixture was stirred, filtered, and washed with ethyl acetate. By concentrating under reduced pressure, a brown amorphous compound 2 (7.43 g) was obtained.

Example 3 Synthesis of Compound 4-1
Compound 2 (760 mg) was dissolved in 30 mL of ethanol, and sodium borohydride (120 mg) was added. After stirring at room temperature for 3 hours, 6N hydrochloric acid (2 mL) was added, and then water (15 mL) was added. The acidity of the reaction system was confirmed with pH test paper (about pH 1), and the mixture was stirred at room temperature overnight. The reaction solution was neutralized with a 5% aqueous sodium hydrogen carbonate solution, added with 100 mL of an aqueous sodium hydrogen carbonate solution, and extracted with 200 mL of ethyl acetate. The ethyl acetate layer was washed with saturated saline and dried over anhydrous sodium sulfate. The organic layer was concentrated to dryness using a rotary evaporator to obtain a brown wax-like compound 4 (600 mg). A part of the obtained target product was further purified by a silica gel column and subjected to NMR measurement. ¹ H-NMR (500 MHz, CD3OD) δ 0.88-0.91 (m, 6H), 1.23-1.37 (m), 1.38-1.43 (m, 2H), 1.51- 1.58 (m, 1H), 1.68-1.75 (m, 1H), 1.96-2.15 (m8H), 3.63 (dd, J = 4.0, 11.5, 1H) ), 3.78 (dd, J = 5.5, 11.0), 3.83-3.86 (m, 1H), 3.99 (dd, J = 3.8, 7.8, 1H). , 4.07-4.09 (m, 1H), 5.36-5.52 (m, 3H), 5.7-5.75 (m, 1H).

¹³ C-NMR (125 MHz, CD3OD) δ 14.46, 23.74, 26.18, 27.89, 28.27, 30.43, 30.47, 30.79, 30.87, 33.08, 33 66, 35.89, 56.03, 61.96, 73.01, 73.27, 129.97, 131.38, 131.48, 134.22, 177.18.

Example 4 Synthesis of Compound 4-2
Compound 2 (1.00 g) was dissolved in ethanol (35.6 mL) under a stream of argon, and sodium borohydride (138 mg) was added at room temperature. After stirring at room temperature for 3 hours, 6N hydrochloric acid (2.44 mL) was added, and after 2 hours, water (18 mL) was added. After stirring at room temperature for 14 hours, the mixture was neutralized with a 6N aqueous sodium hydroxide solution (using 2.1 mL), and a 5% aqueous sodium hydrogen carbonate solution (4 mL) was further added. (PH = 8.1) The solution was concentrated at 50 ° C. under reduced pressure to 22.4 g, and extracted with ethyl acetate (18 mL × 2). The organic layer was washed with 10% saline (18 mL), anhydrous sodium sulfate (1.9 g) was added, and the mixture was stirred, filtered, and washed with ethyl acetate. The filtrate was concentrated under reduced pressure at 50 ° C., dissolved in ethanol (10 mL), and concentrated again under reduced pressure to obtain a brown wax, compound 4 (719 mg).

Example 5 Synthesis of Compound 4-3
Compound 2 (1.00 g) was dissolved in ethanol (18 mL) under a stream of argon, and sodium borohydride (138 mg) was added at room temperature. After stirring at room temperature for 3 hours, 6N hydrochloric acid (2.44 mL) was added, and the mixture was stirred for 20 hours. Neutralized with 6N aqueous sodium hydroxide solution (2.0 mL used, pH = 8.1), and diluted with water (9 mL). Further, a 5% aqueous sodium hydrogen carbonate solution (7 mL) and water (38 mL) were added, and the mixture was extracted with ethyl acetate (54 mL × 2). The organic layer was washed with 10% saline (54 mL), anhydrous sodium sulfate (11.0 g) was added, and the mixture was stirred, filtered, and washed with ethyl acetate. The filtrate was concentrated under reduced pressure at 50 ° C., dissolved in ethanol (10 mL), and again concentrated under reduced pressure to obtain a brown wax, compound 4 (673 mg).

[Example 6] Synthesis of compound 4-4
Compound 2 (1.00 g) was dissolved in ethanol (18 mL) under a stream of argon, and sodium borohydride (138 mg) was added at room temperature. After stirring at room temperature for 3 hours, 6N hydrochloric acid (2.44 mL) was added, and after 1 hour, water (9 mL) was added. After stirring at room temperature for 22 hours, the mixture was neutralized with a 6N aqueous sodium hydroxide solution (use 2.0 mL, pH = 8.8), added with water (45 mL), and extracted with ethyl acetate (54 mL × 2). The organic layer was washed with 10% saline (54 mL), anhydrous sodium sulfate (22.0 g) was added, and the mixture was stirred, filtered, and washed with ethyl acetate. The filtrate was concentrated under reduced pressure at 50 ° C., dissolved in ethanol (10 mL), and concentrated again under reduced pressure to obtain a brown wax, compound 4 (655 mg).

[Example 7]
Hydrous glucosylceramide (64.0 g, solid content 10.9%) was suspended in water (48 mL) and ethanol (70 mL). Sodium periodate (11.6 g) was added over 5 minutes at 29-30 <0> C and stirred at room temperature for 23 hours. After adding sodium L-ascorbate (6.38 g, 32.2 mmol) / city water (10 mL) at 24 to 30 ° C. over 15 minutes, the mixture was concentrated at 50 ° C. under reduced pressure to 133 g. Water (150 mL) and ethyl acetate (100 mL) were added, the mixture was separated, and the aqueous layer was extracted with ethyl acetate (100 mL). The organic layers were combined, anhydrous sodium sulfate (10.0 g) was added, and the mixture was stirred, filtered, and washed with ethyl acetate. The filtrate was concentrated under reduced pressure at 50 ° C. to 19.5 g. Ethanol (67 mL) was added, and the mixture was concentrated under reduced pressure to 19.4 g. Ethanol (67 mL) was further added, and the mixture was concentrated under reduced pressure to obtain a compound 2 solution of 19.1 g. Ethanol (92.3 g) was added to the solution of compound 2 (equivalent to a total of 134 mL of ethanol), and sodium borohydride (1.03 g) was added at 18 to 30 ° C over 4 minutes. After stirring at 22 to 30 ° C. for 3 hours, 6N hydrochloric acid (18.1 mL) was added (22 ° C. → 27 ° C.), and the mixture was stirred for 21 hours. After diluting with water (69 mL) and neutralizing with a 6N aqueous sodium hydroxide solution (use 14.2 mL, pH = 6.0), a 5% aqueous sodium hydrogen carbonate solution (20 mL, pH = 8.0) was added, and the mixture was added with 50 mL of water. The mixture was concentrated under reduced pressure at 123C to 123 g. After two extractions with ethyl acetate (134 mL) (Hartz development), the combined organic layers were washed with 10% saline (134 mL). The organic layer was added with anhydrous sodium sulfate (13.4 g), stirred, filtered, and the filtrate washed with ethyl acetate was concentrated at 50 ° C under reduced pressure to 5.74 g, and then added to ethanol (77 mL) at 50 ° C. And concentrated again under reduced pressure to obtain 5.28 g of a brown wax, compound 4. The obtained compound was analyzed by HPLC (column: Inertsil SIL-100A, 5 μm, 4.6 × 150 mm; A: CHCl ₃ , B: 95% CH ₃ OH; Gradient: 1% to 25%, 15 minutes).

  FIG. 2 shows the result of HPLC analysis of the compound 4 obtained in Example 7. The peak at a retention time (RT) of 8.429 minutes was a mixture of the ceramide represented by the formula (I), and its structure was confirmed using NMR.

[Example 8] Epidermal barrier improvement test using plant-derived ceramide Compound 4 prepared in Example 7 was used as a plant-derived ceramide. Samples 1 to 3 having a composition shown in Table 1 below for an epidermal barrier improvement test were prepared.

Method Samples 1 to 3 (50 μL) were added to the insert of a three-dimensional skin model (Labocyte Epimodel, manufactured by J-TEC), and after culturing for 3 days, the transepidermal water loss (TEWL: Trans Epidermal Water Loss) of each well. ) Was measured using a Tetro TW24 (manufactured by Integral Co., Ltd.), and the ratio to the TEWL value of Sample 1 as a base was calculated.

Result As a result of this calculation, assuming that sample 1 is 100, sample 2 was 96.8 and sample 3 was 83.6. Only in Sample 3 containing plant-derived ceramide, a decrease in TEWL value was observed, suggesting that plant-derived ceramide has an effect of improving the epidermal barrier.

Claims (5)

The following formula (I):

(Wherein R ¹ , R ² , R ³ and R ⁴ independently represent a hydrogen atom, a methyl group, an ethyl group or an acetyl group, and the wavy line indicates that the adjacent double bond is in the E or Z configuration. And n represents an integer of any of 1 to 11.)
Ceramide represented by Wherein ^{R 1,} R 2, ^{R 3} and ^{R 4,} ceramide according to claim 1 is a hydrogen atom.   3. The composition comprising a plurality of ceramides according to claim 1 or 2, wherein the chain length of the fatty acid moiety due to the change in the number of n in the ceramide mixture and the ratio of the arrangement of E or Z are rice ceramides. Ceramide compositions derived from rice, related to their abundance in water.   A skin moisturizer comprising the ceramide according to claim 1 or 2 or the ceramide composition according to claim 3 as an active ingredient. It is a manufacturing method of the ceramide composition of Claim 3, Comprising:
Prepare glucosylceramide extracted from rice,
Oxidizing the glucosylceramide to an aldehyde intermediate by reacting the glucosylceramide and sodium periodate in an aqueous medium containing an organic solvent miscible with water,
In the water-miscible organic solvent, reducing the aldehyde by reacting the aldehyde intermediate and sodium borohydride,
An acid and water are added to the reduction reaction product of the aldehyde intermediate to carry out a hydrolysis reaction, and the following formula (II):

(In the formula, a wavy line indicates that an adjacent double bond has an E or Z configuration, and n represents an integer of any of 1 to 11).
A method for producing a ceramide composition, comprising:

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