Cosmetic composition

Abstract

The present invention provides a composition, such as a cosmetic composition, that contains two or more phytoceramides (PHCs) having different alkyl chain lengths, for example in specific amounts.

Description

[Technical Field]

[0001] This invention relates to compositions such as cosmetic compositions. [Background technology]

[0002] Ceramides are compounds having a structure in which a sphingoid base and a fatty acid are covalently bonded by an amide bond. In other words, ceramides contain a sphingoid base moiety (i.e., an alkyl chain) and a fatty acid moiety (i.e., an acyl chain), and these moieties are covalently bonded to each other by an amide bond. Various groups of ceramides are known, depending on the type of sphingoid base and fatty acid (Non-Patent Literature 1). Some ceramides are used as ingredients in pharmaceuticals and cosmetics.

[0003] Phytoceramide (PHC) is a ceramide of phytosphingosine (PHS). In other words, PHC is a ceramide whose sphingoid base is PHS. It is a mid. That is, a PHC contains a PHS moiety (i.e., an alkyl chain) and a fatty acid moiety (i.e., an acyl chain), and these moieties are covalently bonded to each other by an amide bond. As for PHCs, Various alkyl chain lengths and acyl chain lengths of variant species are known (Non-Patent Literature 1). Compositions containing two or more PHC species with different acyl chain lengths have been reported. (Patent Document 1 and Non-Patent Document 2). However, no compositions containing two or more PHC species with different alkyl chain lengths have been reported. [Prior art documents] [Patent Documents]

[0004] [Patent Document 1] WO2008 / 043386 [Non-patent literature]

[0005] [Non-Patent Document 1] Masukawa Y. et al., J Lipid Res. 2008 Jul;49(7):1466-76. Characterization of overall ceramide species in human stratum corneum. [Non-Patent Document 2] Oh MJ. et al., Clin Cosmet Investig Dermatol. 2017 Sep 13;10:363-371. Novel phytoceramides containing fatty acids of diverse chain lengths are better than a single C18-ceramide N-stearoyl phytosphingosine to improve the physiological properties of human stratum corneum. [Overview of the project] [Problems that the invention aims to solve]

[0006] The object of this invention is to provide compositions such as cosmetic compositions. [Means for solving the problem]

[0007] The inventors have diligently studied to solve the above problems and have found that by using two or more PHC species with different alkyl chain lengths in combination, the properties of compositions containing them can be improved. We discovered this and completed the present invention.

[0008] In other words, the present invention can be illustrated as follows. [1] A composition containing the following ingredient (I): (I) Two or more phytoceramides with different alkyl chain lengths. [2] The aforementioned component (I) is the following phytoceramides (A), (B), and (C): (A) A phytoceramide having a C16 phytosphingosine moiety in an amount of X% by weight relative to the total amount of phytoceramides (A), (B), and (C); (B) A phytoceramide having a C18 phytosphingosine moiety in an amount of Y by weight % relative to the total amount of phytoceramides (A), (B), and (C); (C) A phytoceramide having a C20 phytosphingosine moiety in an amount of Z weight% relative to the total amount of phytoceramides (A), (B), and (C); It consists of, The composition wherein two or all of X, Y, and Z are greater than 0, and all of X, Y, and Z are less than 100. [3] The composition is a cosmetic composition or a pharmaceutical composition. [4] Furthermore, the composition contains ingredients for cosmetic or pharmaceutical use. [5] The composition wherein the cosmetic or pharmaceutical ingredients are selected from the group consisting of excipients, binders, disintegrants, lubricants, stabilizers, diluents, surfactants, pH adjusters, vitamins, minerals, fragrances, pigments, preservatives, terpenoids, lipids, fatty acids, alcohols, water, antioxidants, anti-inflammatory agents, moisturizers, anti-aging agents, anti-cellulite agents, skin whitening agents, skin tanning agents, UV filters, and combinations thereof. [6] The composition comprising at least a terpenoid, a lipid, a fatty acid, or a combination thereof. [7] The composition wherein the terpenoid is selected from the group consisting of cholesterol, cholesterol sulfate, squalene, pristane, and combinations thereof. [8] The composition wherein the lipid is selected from the group consisting of triolein, phosphatidylethanolamine, and combinations thereof. [9] The composition wherein the fatty acid is selected from the group consisting of lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid, oleic acid, linolenic acid, and combinations thereof.

[10] The composition wherein X is from 0 to 90, Y is from 0 to 95, and Z is from 0 to 50.

[11] The composition wherein X is from 65 to 90, Y is from 5 to 30, and Z is from 1 to 20.

[12] The phytoceramide (A) contains phytoceramide having a C16:0 phytosphingosine moiety and the phytoceramide (B) contains phytoceramide having a C18:0 phytosphingosine moiety and the phytoceramide (C) contains phytoceramide having a C20:0 phytosphingosine moiety of the composition.

[13] The phytoceramide (A) has an acyl chain having a length selected from C14 - C26, the phytoceramide (B) has an acyl chain having a length selected from C14 - C26, and the phytoceramide (C) has an acyl chain having a length selected from C14 - C26 of the composition.

Brief Description of the Drawings

[0009] [Figure 1] A figure showing the measurement results of transepidermal water loss (TEWL).

Modes for Carrying Out the Invention

[0010] Hereinafter, the present invention will be described in detail.

[0011] <1> The composition of the present invention The composition of the present invention contains two or more phytoceramides (PHCs) with different alkyl chain lengths. These phytoceramides are collectively referred to as "active ingredients."

[0012] The composition of the present invention may be, for example, a cosmetic composition or a pharmaceutical composition. In particular, the composition of the present invention may be a cosmetic composition. A cosmetic composition refers to a composition used for cosmetic purposes and may be synonymous with cosmetics. A pharmaceutical composition refers to a composition used for pharmaceutical purposes and may be synonymous with pharmaceuticals. Unless otherwise specified, "cosmetic composition" and "pharmaceutical composition" may include quasi-drugs such as medicated cosmetics.

[0013] The compositions of the present invention can be used, for example, by applying them to living organisms. "Application of the compositions of the present invention to living organisms" may mean bringing the compositions of the present invention into contact with a desired part of the living organism to obtain the desired effect. Specifically, the compositions of the present invention can be applied to living organisms in the manner described in the methods of the present invention, which will be described later. The target organisms and application locations of the compositions of the present invention can be appropriately selected according to various conditions such as the purpose of use of the compositions of the present invention. Examples of living organisms (i.e., targets of the compositions of the present invention) include mammals. Examples of mammals include primates such as humans, monkeys, and chimpanzees, rodents such as mice, rats, hamsters, and guinea pigs, and various other mammals such as rabbits, horses, cows, sheep, goats, pigs, dogs, and cats. Humans are particularly noteworthy among mammals. Examples of application locations for the compositions of the present invention include the body surface of living organisms. Examples of body surfaces include skin and body hair. Skin is particularly noteworthy among body surfaces. The skin may be from any part of the body. Examples of skin include the face, head, and body. Examples of body parts include the hands, arms, feet, legs, shoulders, chest, waist, buttocks, back, and any other part. Body hair can come from any part of the body. Examples of body hair include hair on the head.

[0014] The organism may be a healthy individual or an unhealthy individual. The organism may be an individual in which deterioration of the condition of its body surface, such as skin or fur, is observed, or it may not be. Specifically, the organism may be an individual exhibiting symptoms related to deterioration of the condition of its body surface, such as skin or fur, or it may not be.

[0015] The composition of the present invention may be applied to healthy areas or unhealthy areas. The composition of the present invention may be applied to, for example, areas on the body surface such as skin or body hair where deterioration of the condition is observed, or it may not be. Specifically, the composition of the present invention may be applied to, for example, areas on the body surface such as skin or body hair where symptoms related to deterioration of the condition are present, or it may not be.

[0016] The "desired effect" in using the composition of the present invention includes the effect obtained by using the active ingredient (e.g., application to living organisms). That is, the effect obtained by using the composition of the present invention (e.g., application to living organisms) may be obtained by using the active ingredient (e.g., application to living organisms). In other words, the composition of the present invention may have the function of achieving the effect obtained by using the active ingredient (e.g., application to living organisms). The above-mentioned description of the use of the composition of the present invention (e.g., application to living organisms) can be applied mutatis mutandis to the use of the active ingredient (e.g., application to living organisms).

[0017] By using the active ingredient, specifically by applying the active ingredient to an organism, the condition of the application site of the active ingredient in the organism can be improved; that is, an effect of improving the condition of the application site of the active ingredient in the organism can be obtained. Therefore, by using the composition of the present invention, specifically by applying the composition of the present invention to an organism, the condition of the application site of the active ingredient (specifically, the application site of the composition of the present invention) in the organism can be improved; that is, an effect of improving the condition of the application site of the active ingredient (specifically, the application site of the composition of the present invention) in the organism can be obtained. In other words, the composition of the present invention may have the function of improving the condition of the application site of the active ingredient (specifically, the application site of the composition of the present invention). Therefore, the composition of the present invention may be a composition for improving the condition of an application site, for example, a cosmetic or pharmaceutical composition for improving the condition of an application site.

[0018] Improvements in condition include improvements in the condition of the body surface, such as the skin and body hair. Improvements in condition include, in particular, improvements in the condition of the skin. Therefore, the composition of the present invention may be a composition for improving the condition of the body surface, such as the skin and body hair, for example, a cosmetic or pharmaceutical composition for improving the condition of the body surface, such as the skin and body hair. The composition of the present invention may be a composition for improving the condition of the skin, for example, a cosmetic or pharmaceutical composition for improving the condition of the skin.

[0019] Improvements in skin condition include improvements in the skin's lamellar structure (e.g., increased orderliness of the skin's lamellar structure), improvements in the skin's barrier function (e.g., increased function), and improvements in the skin's moisturizing properties (e.g., increased moisture retention). Therefore, the composition of the present invention may be a composition for improving the skin's lamellar structure, improving the skin's barrier function, or improving the skin's moisturizing properties, for example, a cosmetic or pharmaceutical composition for improving the skin's lamellar structure, improving the skin's barrier function, or improving the skin's moisturizing properties. Only one of these indicators may be improved, or two or more may be improved. Furthermore, improvement in one indicator may lead to improvement in another indicator. For example, improvement in the skin's lamellar structure may improve skin functions such as the skin's barrier function and skin moisturizing properties. In other words, a composition for improving the skin's barrier function or skin moisturizing properties may be one embodiment of a composition for improving the skin's lamellar structure. Furthermore, for example, improvement in the skin's barrier function may lead to improved skin moisturizing properties. In other words, a composition for improving skin moisturizing properties may be one embodiment of a composition for improving the skin's barrier function. Specifically, the condition of the skin refers to the condition of the stratum corneum. In other words, improvement of the skin's lamellar structure may specifically refer to improvement of the lamellar structure of the stratum corneum. Furthermore, improvement of the skin's barrier function may specifically refer to improvement of the barrier function of the stratum corneum. Furthermore, improvement of the skin's moisturizing properties may specifically refer to improvement of the moisturizing properties of the stratum corneum.

[0020] There are no particular limitations on the methods used to measure improvements in the condition of the body surface, such as skin and body hair. Improvements in the condition of the body surface can be measured using parameters that reflect the condition of the body surface. For example, a parameter that reflects the condition of the skin is transepidermal water loss. Examples include loss (TEWL) and the order parameter S. That is, improvement in skin condition can be measured, for example, as a decrease in TEWL. TEWL can be measured by conventional methods. Specifically, a decrease in TEWL may mean, for example, an improvement in the skin's barrier function. Also, specifically, a decrease in TEWL may mean, for example, an improvement in the skin's moisturizing ability. Furthermore, improvement in skin condition can be measured, for example, as an increase in the order parameter S. The order parameter can be measured by conventional methods. The order parameter can be measured, for example, by electron spin resonance (ESR) spectroscopy. Specifically, the order parameter can be measured, for example, as described in the examples. The order parameter represents the orderliness of the lipid chain arrangement (e.g., the orderliness of the lamellar structure), and a higher order parameter S means that the lipid chains are more neatly arranged. Specifically, an increase in the order parameter S may mean, for example, an improvement in the lamellar structure of the skin.

[0021] The indicators exemplified above all only need to show improvement when the active ingredient is used (specifically, when the composition of the present invention is used) compared to when the active ingredient is not used (specifically, when the composition of the present invention is not used). Furthermore, "improvement" for the indicators exemplified above includes, in cases where the indicator deteriorates when the active ingredient is not used (specifically, when the composition of the present invention is not used), a reduction in such deterioration when the active ingredient is used (specifically, when the composition of the present invention is used).

[0022] By using an active ingredient, specifically by applying the active ingredient to a living organism, an effect based on improvement of the condition of the application site of the active ingredient may be obtained. By improving the condition of the application site of the active ingredient, it is expected that symptoms related to the deterioration of the condition of that site can be prevented and / or treated. That is, an effect based on improvement of the condition of the application site of the active ingredient is the effect of preventing and / or treating symptoms related to the deterioration of the condition of that site. Symptoms related to the deterioration of the condition of the application site of the active ingredient may be diseases or not. Symptoms related to the deterioration of the condition of the application site of the active ingredient include symptoms caused by the deterioration of the condition of that site and symptoms accompanied by the deterioration of the condition of that site. Therefore, the composition of the present invention may be a composition for the prevention and / or treatment of symptoms related to the deterioration of the condition of the application site, for example, a cosmetic or pharmaceutical composition for the prevention and / or treatment of symptoms related to the deterioration of the condition of the application site.

[0023] Examples of deterioration include deterioration of the condition of the body surface, such as the skin and body hair. In particular, deterioration of the skin condition is a notable example. Therefore, the composition of the present invention may be a composition for preventing and / or treating symptoms related to deterioration of the condition of the body surface, such as the skin and body hair, for example, a cosmetic or pharmaceutical composition for preventing and / or treating symptoms related to deterioration of the condition of the body surface, such as the skin and body hair. In particular, the composition of the present invention may be a composition for preventing and / or treating symptoms related to deterioration of the skin condition, for example, a cosmetic or pharmaceutical composition for preventing and / or treating symptoms related to deterioration of the skin condition.

[0024] Deterioration of skin condition can include deterioration of the skin's lamellar structure (e.g., decreased order in the skin's lamellar structure), deterioration of the skin's barrier function (e.g., decreased function), and deterioration of the skin's moisture retention (e.g., decreased moisture retention). Symptoms associated with deterioration of skin condition include rough skin, dry skin, scaling, desquamation, dermatitis, and psoriasis.

[0025] The active ingredient may be used for therapeutic purposes or for non-therapeutic purposes. That is, unless otherwise specified, the effects exemplified above may be obtained for therapeutic purposes or for non-therapeutic purposes. In the case of non-therapeutic purposes, "treatment of symptoms" may be interpreted as "improvement of symptoms." "Therapeutic purposes" may mean, for example, procedures performed on the human body for therapeutic purposes, and may particularly mean that the procedure is performed as a medical procedure. "Non-therapeutic purposes" may mean, for example, that the procedure does not include procedures performed on the human body for therapeutic purposes, and may particularly mean that the procedure is performed as a non-medical procedure. Examples of non-therapeutic purposes include health promotion and cosmetic purposes.

[0026] By using a combination of active ingredients, the properties of the composition can be improved compared to using any one of the active ingredients alone; in other words, an effect of improving the properties of the composition can be obtained. This effect is also called the "combination effect of active ingredients." For example, the combination effect of active ingredients can be obtained in the composition of the present invention. Properties include physical properties, chemical properties, and physiological properties. Specifically, improvements in properties include a decrease in the melting point of the composition and an increase in the lamellar structure-forming ability of the composition. Specifically, improvements in properties also include an increase in the function that provides the effects obtained by using the active ingredients. Such functions include a function that improves the condition of the application site of the active ingredient (specifically, the application site of the composition of the present invention) and a function that prevents and / or treats symptoms related to the deterioration of the condition of the application site of the active ingredient (specifically, the application site of the composition of the present invention). An increase in the lamellar structure-forming ability of the composition can be measured, for example, as an increase in the appearance of a Maltese cross pattern. A Maltese cross pattern can be observed as described in the examples. The combination of active ingredients may provide only one of these effects, or two or more of these effects. One effect may result in another. For example, an increase in lamellar structure formation ability may enhance the function that provides the effect obtained by using the active ingredient.

[0027] Phytoceramide (PHC) is a ceramide derived from phytosphingosine (PHS). PHC is also known as "ceramide 3" or "ceramide NP." Each variation of PHC is also called a "PHC species," and each variation of PHS is also called a "PHS species."

[0028] "Phytosphingosine (PHS)" is a long-chain amino acid called a sphingoid base. This refers to an alcohol having the following structure. PHS has an amino group at the C2 position. The alkyl chain includes an alkyl chain. That is, the carbon at one end of the alkyl chain that is bonded to the aminated carbon (C2 position) is considered to be the C1 position of the alkyl chain. The alkyl chain has two or more hydroxyl groups. The alkyl chain may have hydroxyl groups at positions C1, C3, and C4, for example. The alkyl chain may have additional hydroxyl groups other than at positions C1, C3, and C4, or it may not. Typically, the alkyl chain does not need to have additional hydroxyl groups other than at positions C1, C3, and C4. The length and degree of unsaturation of the alkyl chain may be variable. The alkyl chain length may be, for example, C14-C26 (C14, C16, C18, C20, C22, C24, C26, etc.). The alkyl chain length may be, for example, C16, C18, or C20 in particular. The alkyl chain length can be interpreted as the number of carbon atoms in the alkyl chain. The alkyl chain may be saturated or unsaturated. The alkyl chain may have one or more unsaturated double bonds. That is, unless otherwise specified, the "alkyl chain" for PHS and PHC is not limited to saturated chains and may include unsaturated chains such as alkenyl chains and alkadienyl chains. Typically, the alkyl chain may not have any unsaturated double bonds, or may have only one unsaturated double bond. More typically, the alkyl chain may not have any unsaturated double bonds. The alkyl chain may have, for example, a C8 trans double bond. The chiral center configuration may be the same as or different from that of the PHS moiety of natural PHCs. The C2 position may be, for example, 2S. The C3 position may be, for example, 3S. The C4 position may be, for example, 4R. The chiral center configuration may be, for example, 2S, 3S, 4R. The number of carbon atoms in the alkyl chain of a PHS can be denoted by "n". A PHS having an alkyl chain with "n" carbon atoms is also called a "Cn PHS" or "Cn alkyl PHS (Cn-alkyl PHS)". For example, "C18 PHS" is a general term for PHS having an alkyl chain of C18 length, which may be saturated or unsaturated. The number of unsaturated double bonds in the alkyl chain of a PHS can be denoted by "m".PHS having an alkyl chain with n carbon atoms and m unsaturated double bonds is also called "Cn:m PHS" or "Cn:m alkyl PHS (Cn:m-alkyl PHS)". Examples of PHS include variant species of such PHS with different lengths and / or degrees of unsaturation. Specifically, examples of PHS variant species include C16:0 PHS having a saturated C16 alkyl chain, C18:0 PHS having a saturated C18 alkyl chain, C20:0 PHS having a saturated C20 alkyl chain, C18:1 PHS having a C18 alkyl chain with one unsaturated double bond, and C20:1 PHS having a C20 alkyl chain with one unsaturated double bond. More specifically, variant species of PHS include C16:0 PHS, C18:0 PHS, C20:0 PHS, C18:1 PHS, and C20:1 PHS, which do not have additional hydroxyl groups other than at the C1, C3, and C4 positions. Variant species of PHS include adducts of PHS, such as 4-(hydroxymethyl)-2-methyl-6-tetradecanyl-1,3-oxazinan-5-ol and 4-(hydroxymethyl)-2-methyl-6-hexadecanyl-1,3-oxazinan-5-ol (these can be produced by the reaction of C18:0 PHS and C20:0 PHS with acetaldehyde, respectively). "Phytosphingosine (PHS)" is not limited to the typical PHS species C18:0 PHS, but may also refer collectively to variant species of PHS such as C16:0 PHS, C18:0 PHS, C20:0 PHS, C18:1 PHS, and C20:1 PHS, and may also refer collectively to such PHS variant species and their accretionary compounds.

[0029] "Phytoceramide (PHC)" refers to a compound having a structure in which PHS is covalently bonded to a fatty acid by an amide bond. In other words, PHC consists of a PHS moiety (i.e., a moiety equivalent to PHS) and a fatty acid moiety. It contains a PHS (i.e., a fatty acid) portion, and these portions are covalently bonded to each other by amide bonds. The PHS portion is also called an "alkyl chain." The fatty acid portion is also called an "acyl chain." The amide bond is formed between the amino group at the C2 position of PHS and the carboxyl group of the fatty acid. The above explanation regarding PHS can be applied mutatis mutandis to the PHS portion of PHC. That is, for example, The length and degree of unsaturation of the chain (i.e., the PHS portion) may be variable, as in the case of PHS. That is, as PHCs, examples of PHS species of ceramide can be found. Specifically, examples include C16 PHS, C18 PHS, and C20 PHS ceramides. More specifically, examples of PHCs include C16:0 PHS, C18:0 PHS, C20:0 PHS, C18:1 PHS, and C20:1 PHS ceramides. The length and degree of unsaturation of the acyl chain (i.e., the fatty acid portion) of the PHC may be variable. The chain length may be, for example, C14 to C26 (C14, C16, C18, C20, C22, C24, C26, etc.). The acyl chain length may be, for example, C18 in particular. The acyl chain length is the number of carbon atoms in the acyl chain. This can be interpreted as the number of carbon atoms. The acyl chain may be saturated or unsaturated. The acyl chain may have one or more unsaturated double bonds. The acyl chain may or may not have functional groups (i.e., substituents). The acyl chain may have one or more functional groups (substituents). A hydroxyl group is an example of a functional group (substituent). For example, the acyl chain may or may not have a hydroxyl group at the C2 position. Typically, the acyl chain does not have a hydroxyl group at the C2 position. Note that the carbon atom constituting the amide bond is considered to be at the C1 position of the acyl chain. Typically, the acyl chain does not have a hydroxyl group. Typically, the acyl chain does not have functional groups (substituents). A PHC having an alkyl chain with n carbon atoms (i.e., a PHC having a Cn PHS moiety) is also called a "Cn alkyl PHC," "Cn PHS (phyto)ceramide," or "Cn PHS (phyto)ceramide." The number of carbon atoms in the acyl chain of a PHC can be denoted by "x". A PHC having an acyl chain with "x" carbon atoms (i.e., a PHC with a Cx acyl moiety) is also called a "Cx acyl PHC". The number of unsaturated double bonds in the acyl chain of a PHC can be denoted by "y". A PHC having an acyl chain with "x" carbon atoms and "y" unsaturated double bonds (i.e., a PHC with a Cx:y acyl moiety) is also called a "Cx:y acyl PHC". A PHC having an alkyl chain with "n" carbon atoms and an acyl chain with "x" carbon atoms (i.e., a PHC with a Cn PHS moiety and a Cx acyl moiety) is also called a "Cn alkyl / Cx acyl PHC". For example, "C18 alkyl PHC", "C18 PHS ceramide", or "C18 PHS ceramide" is a general term for PHC species having an alkyl chain of length C18 and an arbitrary acyl chain. Furthermore, for example, "C14 acyl PHC" is a general term for PHC species having an acyl chain of length C14 and an arbitrary alkyl chain. Also, for example, "C18 alkyl / C14 acyl PHC" is a general term for PHC species having an alkyl chain of length C18 and an acyl chain of length C14.In the case of a PHC having an alkyl chain with n carbon atoms and m unsaturated double bonds (i.e., a PHC having a Cn:m PHS moiety), "Cn" in the above name can be rewritten as "Cn:m". The same applies to "Cx" in the acyl chain. For example, "C18:1 alkyl / C14:0 acyl PHC" is a general term for PHC species having an unsaturated alkyl chain of length C18 with one unsaturated double bond and a saturated acyl chain of length C14. Unless otherwise specified, the PHC specified by the above name may consist of one type of PHC species, or it may consist of a combination of two or more types of PHC species. For example, Cn PHS ceramide (Cn alkyl PHC) may consist of one type of Cn PHS ceramide, or it may consist of a combination of two or more types of Cn PHS ceramide. Such a combination may consist of two or more Cn PHS ceramides having different alkyl chains and / or different acyl chains (e.g., alkyl chains with different degrees of unsaturation and / or acyl chains with different lengths and / or degrees of unsaturation). Alternatively, for example, a Cn:m PHS ceramide may consist of one Cn:m PHS ceramide or a combination of two or more Cn:m PHS ceramides. Such a combination may consist of two or more Cn:m PHS ceramides having different acyl chains (e.g., acyl chains with different lengths and / or degrees of unsaturation).

[0030] The combination of active ingredients and the ratio of their content are not particularly limited, as long as the desired effect is achieved.

[0031] The combination of active ingredients is C16 PHS ceramide, C18 PHS ceramide, and C20 PHS Examples include combinations of two or all ceramides. That is, the active ingredients may be, for example, a combination of C16 PHS ceramide and C18 PHS ceramide, a combination of C16 PHS ceramide and C20 PHS ceramide, a combination of C18 PHS ceramide and C20 PHS ceramide, or C16 PHS ceramide. It may be a combination of C18 PHS ceramide and C20 PHS ceramide.

[0032] Content of each active ingredient (i.e., PHCs with each alkyl chain length) relative to the total amount of active ingredients The quantities are, for example, 0.01% by weight or more, 0.1% by weight or more, 1% by weight or more, 2% by weight or more, 3% by weight or more, 5% by weight or more, 10% by weight or more, 15% by weight or more, 20% by weight or more, 25% by weight or more, 30% by weight It may be % or more, 35% or more by weight, 40% or more by weight, 45% or more by weight, 50% or more by weight, 55% or more by weight, 60% or more by weight, 65% or more by weight, 70% or more by weight, 75% or more by weight, 80% or more by weight, 85% or more by weight, or 90% or more by weight, and it may be less than 100g, 99.99% or less by weight, 99% or less by weight, 97% or less by weight, 95% or less by weight, 90% or less by weight, 85% or less by weight, 80% or less by weight, 75% or less by weight, 70% or less by weight, 65% or less by weight, 60% or less by weight, 55% or less by weight, 50% or less by weight, 45% or less by weight, 40% or less by weight, 35% or less by weight, 30% or less by weight, 25% or less by weight, 20% or less by weight, 15% or less by weight, or 10% or less by weight, and may be within a range of non-contradictory combinations thereof. The content of each active ingredient relative to the total amount of active ingredients may specifically be, for example, 0.01-99.99% by weight, 0.1-99.9% by weight, 1-99% by weight, 5-95% by weight, 10-90% by weight, 15-85% by weight, 20-80% by weight, 25-75% by weight, 1-90% by weight, 5-80% by weight, 10-70% by weight, 15-60% by weight, 20-50% by weight, 25-40% by weight, 1-60% by weight, 5-50% by weight, 10-40% by weight, 15-35% by weight, or 20-30% by weight. The content of each active ingredient relative to the total amount of active ingredients may specifically be, for example, 0.01-10% by weight, 0.1-10% by weight, 1-10% by weight, 10-20% by weight, 20-30% by weight, 30-40% by weight, 40-50% by weight, 50-60% by weight, 60-70% by weight, 70-80% by weight, 80-90% by weight, 90-99% by weight, 90-99.9% by weight, or 90-99.99% by weight.

[0033] In other words, the compositions of the present invention include, for example, the following PHC(A), (B), and (C) The composition may contain: (A) A PHC having an amount of X weight% of the total amount of PHC(A), (B), and (C) (i.e., C16 PHS ceramide); (B) A PHC having a C18 PHS moiety in an amount of Y by weight % relative to the total amount of PHC(A), (B), and (C) (i.e., C18 PHS ceramide); (C) A PHC having an amount of Z weight% of the total amount of PHC(A), (B), and (C) (i.e., C20 PHS ceramide).

[0034] Two or all of X, Y, and Z are greater than 0, and all of X, Y, and Z are less than 100. In other words, only one of X, Y, and Z may be 0. In this case, PHC(A), (B), and (C), specifically those in which the content of PHC(A), (B), and (C) is greater than 0, are considered to be the active ingredients.

[0035] The PHC(A), (B), and (C) content may be, for example, 0% by weight or more, or the content of each of the active ingredients exemplified above, or within a range of non-inconsistent combinations thereof.

[0036] That is, X, Y, and Z are, for example, 0 or more, 1 or more, 2 or more, 3 or more, 5, respectively. X may be 10 or more, 15 or more, 20 or more, 25 or more, 30 or more, 35 or more, 40 or more, 45 or more, 50 or more, 55 or more, 60 or more, 65 or more, 70 or more, 75 or more, 80 or more, 85 or more, or 90 or more, and may be less than 100, 95 or less, 90 or less, 85 or less, 80 or less, 75 or less, 70 or less, 65 or less, 60 or less, 55 or less, 50 or less, 45 or less, 40 or less, 35 or less, 30 or less, 25 or less, 20 or less, 15 or less, or 10 or less, and may be within a range of non-contradictory combinations thereof. Specifically, X may be, for example, 0-90, 1-85, 2-80, 65-90, 70-85, 5-20, or 1-15. Specifically, Y may be, for example, 0-95, 10-90, 15-85, 75-90, 50-65, 5-30, or 10-25. Specifically, Z may be, for example, 0-50, 2-40, 5-35, 25-40, 5-20, 1-20, or 2-15.

[0037] In particular, X may be between 0 and 90, Y may be between 0 and 95, and Z may be between 0 and 50. More specifically, X may be between 1 and 85, Y may be between 10 and 90, and Z may be between 2 and 40. It may also be the case that X is between 2 and 80, and Y is between 15 and 85. Furthermore, Z may be between 5 and 35.

[0038] In another embodiment, X may be 65 to 90, Y may be 5 to 30, and Z may be 1 to 20. It may be present. In yet another embodiment, X may be 70 to 85, Y may be 10 to 25, and Z may be 2 to 15.

[0039] In another embodiment, X may be 1 to 15, Y may be 75 to 90, and Z may be 5 to 20. It's okay to have it.

[0040] In another embodiment, X may be 5 to 20, Y may be 50 to 65, and Z may be 25 to 40.

[0041] Each active ingredient (i.e., PHC having each alkyl chain length) may consist of one PHC species, or it may consist of a combination of two or more PHC species. Each of the one, two, or more PHC species constituting each active ingredient is not particularly limited, as long as it is a PHC species having the corresponding alkyl chain length. This refers to the length corresponding to each active ingredient. Each active ingredient is a set of two or more PHC species. When referring to a combination of ingredients, "content of each active ingredient" means the total amount of that combination.

[0042] Each active ingredient (i.e., PHC having each alkyl chain length) is, for example, the corresponding alkyl It may contain any of the above-exemplified PHC species having a chain length. That is, each active ingredient Insofar as it has the corresponding alkyl chain length, it possesses any one or more of the characteristics of the above PHC. Each active ingredient may have, for example, a PHC species having a corresponding alkyl chain length in which the acyl chain is unsubstituted. Each active ingredient may specifically include, For example, a PHC having the corresponding alkyl chain length, wherein the alkyl chain has hydroxyl groups at the C1, C3, and C4 positions and does not have any additional hydroxyl groups other than those hydroxyl groups. It may contain species. Specifically, each active ingredient may contain, for example, a PHC species having a corresponding alkyl chain length in which the alkyl chain has no or only one unsaturated double bond. i. "The active ingredient (i.e., a PHC having a certain alkyl chain length) contains a certain PHC species" means, This means that at least one PHC species is used as the active ingredient, and this active ingredient is This includes cases where the active ingredient consists of a certain PHC species, or where the active ingredient consists of a combination of the certain PHC species and one or more other PHC species having corresponding alkyl chain lengths. If a component (i.e., a PHC having a certain alkyl chain length) contains a certain PHC species, the ratio of the amount of that PHC species to the total amount of the active ingredient is, for example, 30% by weight or more, 40% by weight or more, 50% by weight or more. The amount may be % by weight or more, 60% by weight or more, 70% by weight or more, 80% by weight or more, 90% by weight or more, 95% by weight or more, 97% by weight or more, 99% by weight or more, 99.5% by weight or more, 99.9% by weight or more, or 100% by weight. Each active ingredient may have an acyl chain of a length selected from C14 to C26 (e.g., C14, C16, C18, C20, C22, C24, and C26). "An active ingredient (i.e., a PHC having a certain alkyl chain length) has an acyl chain of a length selected from C14 to C26" means that if the active ingredient consists of a combination of two or more PHC species, each PHC species independently has an acyl chain of a length selected from C14 to C26 (e.g., C14, C16, C18, C20, C22, C24, and C26).

[0043] PHC(A) may consist of one type of C16 PHS ceramide, or two or more types. It may consist of the above combination of C16 PHS ceramides. PHC(A) may consist of two types or If it consists of more than one combination of C16 PHS ceramides, X represents the total amount of the combination. PHC(A) may contain C16:0 PHS ceramide. "PHC(A) contains C16:0 PHS ceramide" means that at least C16:0 PHS ceramide is used as PHC(A), and may include cases where PHC(A) consists of C16:0 PHS ceramide or a combination of C16:0 PHS ceramide and one or more other C16 PHS ceramides. The ratio of the amount of C16:0 PHS ceramide to the total amount of PHC(A) may be, for example, 30% by weight or more, 40% by weight or more, 50% by weight or more, 60% by weight or more, 70% by weight or more, 80% by weight or more, 90% by weight or more, 95% by weight or more, 97% by weight or more, 99% by weight or more, 99.5% by weight or more, 99.9% by weight or more, or 100%. PHC(A) may have any of the above-described characteristics of a PHC, as long as it has a C16 PHS moiety. For example, PHC(A) may have an acyl chain of a length selected from C14 to C26 (e.g., C14, C16, C18, C20, C22, C24, and C26). "PHC(A) has an acyl chain of a length selected from C14 to C26" means that if PHC(A) consists of a combination of two or more C16 PHS ceramides, each C16 PHS ceramide independently has an acyl chain of a length selected from C14 to C26 (e.g., C14, C16, C18, C20, C22, C24, and C26).

[0044] PHC(B) may consist of one type of C18 PHS ceramide, or two or more types. It may consist of the above combination of C18 PHS ceramides. PHC(B) may consist of two types or If it consists of more than one combination of C18 PHS ceramides, Y represents the total amount of that combination. PHC(B) may contain C18:0 PHS ceramide. "PHC(B) contains C18:0 PHS ceramide" means that at least C18:0 PHS ceramide is used as PHC(B), and may include cases where PHC(B) consists of C18:0 PHS ceramide or a combination of C18:0 PHS ceramide and one or more other C18 PHS ceramides. The ratio of the amount of C18:0 PHS ceramide to the total amount of PHC(B) may be, for example, 30% by weight or more, 40% by weight or more, 50% by weight or more, 60% by weight or more, 70% by weight or more, 80% by weight or more, 90% by weight or more, 95% by weight or more, 97% by weight or more, 99% by weight or more, 99.5% by weight or more, 99.9% by weight or more, or 100%. PHC(B) may have any of the above-described characteristics of a PHC, as long as it has a C18 PHS moiety. For example, PHC(B) may have an acyl chain of a length selected from C14 to C26 (e.g., C14, C16, C18, C20, C22, C24, and C26). "PHC(B) has an acyl chain of a length selected from C14 to C26" means that if PHC(B) consists of a combination of two or more C18 PHS ceramides, each C18 PHS ceramide independently has an acyl chain of a length selected from C14 to C26 (e.g., C14, C16, C18, C20, C22, C24, and C26).

[0045] PHC(C) may consist of one type of C20 PHS ceramide, or two or more types. It may consist of the above C20 PHS ceramide combination. PHC(C) may consist of two types or If it consists of more than one combination of C20 PHS ceramides, Z represents the total amount of that combination. PHC(C) may contain C20:0 PHS ceramide. "PHC(C) contains C20:0 PHS ceramide" means that at least C20:0 PHS ceramide is used as PHC(C), and may include cases where PHC(C) consists of C20:0 PHS ceramide, or a combination of C20:0 PHS ceramide and one or more other C20 PHS ceramides. The ratio of the amount of C20:0 PHS ceramide to the total amount of PHC(C) may be, for example, 30% or more by weight, 40% or more by weight, 50% or more by weight, 60% or more by weight, 70% or more by weight, 80% or more by weight, 90% or more by weight, 95% or more by weight, 97% or more by weight, 99% or more by weight, 99.5% or more by weight, 99.9% or more by weight, or 100%. PHC(C) may have any of the above-described characteristics of a PHC, as long as it has a C20 PHS moiety. For example, PHC(C) may have an acyl chain of a length selected from C14 to C26 (e.g., C14, C16, C18, C20, C22, C24, and C26). "PHC(C) has an acyl chain of a length selected from C14 to C26" means that if PHC(C) consists of a combination of two or more C20 PHS ceramides, each C20 PHS ceramide independently has an acyl chain of a length selected from C14 to C26 (e.g., C14, C16, C18, C20, C22, C24, and C26).

[0046] As for each active ingredient, commercially available PHC may be used, or PHC may be manufactured and obtained as appropriate. The method of manufacturing PHC is not particularly limited. PHC can be manufactured by known methods, for example. PHC can be manufactured by chemical synthesis, enzymatic methods, bioconversion methods, fermentation methods, extraction methods, etc. They can be manufactured by a combination of these. Specifically, PHCs are, for example, yeast It can be produced by a fermentation method using a mother. Alternatively, PHC can be specifically, for example, It can be manufactured by PHS conversion. The manufacturing method for PHC can be independently selected for each active ingredient.

[0047] The method for manufacturing PHS is not particularly limited. PHS can be manufactured, for example, by known methods. PHS can be manufactured, for example, by chemical synthesis, enzymatic methods, biotransformation methods, fermentation methods, extraction methods, or These can be manufactured by combining them. Specifically, PHS uses, for example, yeast It can be manufactured using the fermentation method employed.

[0048] In other words, each active ingredient may be a PHC produced by a fermentation method, such as a fermentation method using yeast. Alternatively, each active ingredient may be produced by a fermentation method, such as a fermentation method using yeast. Therefore, a PHC having a manufactured PHS component may also be used.

[0049] Examples of methods for producing PHC or PHS by fermentation using yeast, and methods for producing PHC by converting PHS, will be described later.

[0050] Each PHC constituting the active ingredient may be manufactured individually, or two or more PHCs constituting the active ingredient may be manufactured together as a mixture. A mixture containing seeds is also called a "PHC mixture." PHC and / or PHC mixtures produced in this manner can be used as active ingredients in appropriate combinations. That is, For example, two or more PHCs may be combined with each other, or one or more PHCs may be combined with one or more PHC mixtures, or two or more The PHC mixtures may be combined with each other, thereby forming a combination of active ingredients. Alternatively, the PHC mixture may be used as is as the active ingredient.

[0051] As for each active ingredient, PHC that has been purified to a desired degree may be used, or a material containing PHC may be used.

[0052] The composition of the present invention may consist of an active ingredient, or it may contain ingredients other than the active ingredient. That is, the active ingredient may be used as is or in combination with other ingredients as part of the composition of the present invention. The ingredients other than the active ingredient may be one ingredient, or two or more ingredients may be used in combination.

[0053] Other components are not particularly limited as long as the desired effect is obtained. Other components may be acceptable depending on the intended use of the composition of the present invention. Examples of other components include physiologically acceptable components. Examples of physiologically acceptable components include cosmetic and pharmaceutical components, such as those commonly used in cosmetic and pharmaceutical applications. "Cosmetic components" refers to components that can be used as cosmetic components. "Pharmaceutical components" refers to components that can be used as pharmaceutical components. Specific examples of such components include excipients, binders, disintegrants, lubricants, stabilizers, diluents, surfactants, pH adjusters, vitamins, minerals, fragrances, pigments, preservatives, terpenoids, lipids, fatty acids, alcohols, and water. Other components include terpenoids, lipids, and fatty acids in particular. Examples of terpenoids include sterols, squalene, and pristane. Examples of sterols include cholesterol and cholesterol sulfate. Examples of lipids include triacylglycerols and glycerophospholipids. Examples of triacylglycerols include triolein. Examples of glycerophospholipids include phosphatidylethanolamine. Examples of fatty acids include lauric acid (12:0), myristic acid (14:0), palmitic acid (16:0), stearic acid (18:0), arachidic acid (20:0), behenic acid (22:0), lignoceric acid (24:0), cerotic acid (26:0), myristoleic acid (14:1), palmitoleic acid (16:1), oleic acid (18:1), linoleic acid (18:2), and linolenic acid (18:3). In particular, examples of fatty acids include lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid, oleic acid, and linolenic acid. Fatty acids include, more specifically, lauric acid, palmitic acid, stearic acid, arachidic acid, behenic acid, and lignoceric acid. Lauric acid is a particularly notable fatty acid. Other such components include physiologically active substances such as antioxidants, anti-inflammatory agents, moisturizers, anti-aging agents, anti-cellulite agents, skin whitening agents, skin sunscreens, and UV filters.Furthermore, other components that were not selected as active ingredients include ceramides such as PHC. As components other than active ingredients, for example, at least one or more of the components exemplified above may be selected. In other words, components other than active ingredients may include, for example, one or more of the components exemplified above. "A certain component is selected as a component other than an active ingredient" or "a component other than an active ingredient includes a certain component" means that at least that certain component is used as a component other than an active ingredient, and includes cases where the component other than an active ingredient consists of that certain component, or where the component other than an active ingredient consists of that certain component and one or more other components. Any of the components exemplified above can be used, for example, as ingredients for cosmetics and ingredients for pharmaceuticals.

[0054] If components other than the active ingredient can take the form of a salt, these components may be in the form of a free form, a salt thereof, or a mixture thereof. For example, salts with acidic groups such as carboxyl groups (e.g., salts of fatty acids) include ammonium salts; salts with alkali metals such as sodium and potassium; salts with alkaline earth metals such as calcium and magnesium; and salts with other metals such as aluminum and zinc.

[0055] The shape of the composition of the present invention is not particularly limited as long as the desired effect is obtained. The shape of the composition of the present invention can be set according to various conditions such as the type of active ingredient, the types and amounts of other ingredients, the intended use of the composition of the present invention, and the manner in which the composition of the present invention is used. The composition of the present invention may be formulated into a desired shape. The composition of the present invention may be provided, for example, in the form of any cosmetic composition or pharmaceutical composition that can be applied to the skin. Examples of cosmetic compositions or pharmaceutical compositions (especially cosmetic compositions) include skincare cosmetics, makeup cosmetics, point makeup cosmetics, hair care products, hair styling products, hair dyes, bleaching agents, permanent wave agents, waving agents, body care cosmetics, and UV care cosmetics. Examples of skincare cosmetics include cleansing cosmetics (facial cleansers, makeup removers, etc.), lotions, serums, packs, massage creams, emulsions, creams, and essences. Examples of makeup cosmetics include foundations, concealers, face powders, and makeup bases. Examples of point makeup cosmetics include lipsticks, blushes, and eye makeup (eyeshadows, eyeliners, mascaras, eyebrow products, etc.). Examples of hair care products include shampoos and conditioners (also called rinses or treatments). Hair care products may, for example, be used to obtain the effects described above on the hair and / or scalp. Examples of hair styling products include hair creams, hair waxes, hair liquids, and hair sprays. Examples of hair dyes or bleaches include hair manicures and hair colors. Examples of body care cosmetics include body creams, soaps, body washes, hand soaps, and antiperspirants. Examples of UV care cosmetics include sunscreens, suntan cosmetics, self-tanning cosmetics, and after-sun cosmetics. Examples of cosmetic compositions or pharmaceutical compositions (especially pharmaceutical compositions) include topical preparations such as skin preparations. Examples of topical preparations include solid preparations (powder, etc.), liquid preparations (lotions, liniments, etc.), sprays, ointments, creams, gels, and patches (tapes, poultices, etc.). The compositions of the present invention may be provided in a form that can be applied directly to the skin, or they may be prepared into a form that can be applied to the skin before use.

[0056] The content of the components (i.e., the active ingredient and optionally other components) in the composition of the present invention is not particularly limited, as long as the desired effect is obtained. The content of the components in the composition of the present invention can be set according to various conditions such as the type of active ingredient, the types of other components, the shape of the composition of the present invention, the intended use of the composition of the present invention, and the manner in which the composition of the present invention is used.

[0057] The total content of the active ingredient in the composition of the present invention is, for example, 0.01% by weight or more, and 0.1% by weight or less. The above may be 1% or more by weight, 5% or more by weight, or 10% or more by weight, and may be 100% or less by weight, or 99.9% or less by weight. Less than or equal to % by weight, less than or equal to 70% by weight, less than or equal to 50% by weight, less than or equal to 30% by weight, less than or equal to 10% by weight, less than or equal to 5% by weight, and The amount may be 1% by weight or less, and may be within a range of non-inconsistent combinations thereof. Specifically, the total content of the active ingredients in the composition of the present invention may be, for example, 0.01 to 1% by weight, 1 to 5% by weight, 5 to 10% by weight, 10 to 20% by weight, 20 to 30% by weight, or 30 to 50% by weight. Specifically, the total content of the active ingredients in the composition of the present invention may be, for example, 0.01 to 50% by weight, 0.01 to 30% by weight, 0.01 to 10% by weight, or 0.01 to 5% by weight. Specifically, the total content of the active ingredients in the composition of the present invention may be, for example, 1 to 50% by weight, 1 to 30% by weight, 1 to 10% by weight, or 1 to 5% by weight. Specifically, the total content of the active ingredients in the composition of the present invention may be, for example, 5 to 50% by weight, 5 to 30% by weight, or 5 to 10% by weight.

[0058] If the composition of the present invention contains ceramides other than the active ingredient, the ratio of the total content of the active ingredient to the total content of ceramides in the composition of the present invention may be, for example, 30% by weight or more, 40% by weight or more, 50% by weight or more, 60% by weight or more, 70% by weight or more, 80% by weight or more, 90% by weight or more, 95% by weight or more, 97% by weight or more, 99% by weight or more, 99.5% by weight or more, or 99.9% by weight or more.

[0059] Furthermore, the total amount of the active ingredient in the composition of the present invention may be such that, for example, the total amount of the active ingredient applied when the composition of the present invention is applied to a body surface such as skin or body hair falls within a desired range.

[0060] Furthermore, the content of each active ingredient shall be calculated based on the amount of the active ingredient itself in the material, in cases where a material containing the active ingredient is used.

[0061] <2> Method of the present invention The method of the present invention is a method that includes the step of applying the composition of the present invention to a living organism.

[0062] By applying the composition of the present invention to an organism using the method of the present invention, the condition of the application site of the composition of the present invention in the organism may be improved; that is, an effect of improving the condition of the application site of the composition of the present invention in the organism may be obtained. In other words, the method of the present invention may be a method for improving the condition of the application site of the composition of the present invention.

[0063] The improvement of the condition is as described above. That is, the method of the present invention may be a method for improving the condition of the body surface, such as the skin and body hair. In particular, the method of the present invention may be a method for improving the condition of the skin. The method of the present invention may be a method for improving the lamellar structure of the skin, improving the barrier function of the skin, or improving the moisturizing properties of the skin. A method for improving the barrier function of the skin or improving the moisturizing properties of the skin may be one embodiment of a method for improving the lamellar structure of the skin. Furthermore, a method for improving the moisturizing properties of the skin may be one embodiment of a method for improving the barrier function of the skin.

[0064] By applying the composition of the present invention to an organism using the method of the present invention, an effect based on improvement of the condition of the application site of the composition of the present invention may be obtained in the organism. In other words, the method of the present invention may be a method for obtaining an effect based on improvement of the condition of the application site of the composition of the present invention.

[0065] The effects based on the improvement of the condition of the application site of the composition of the present invention (i.e., the application site of the active ingredient) are as described above. That is, the method of the present invention may be a method for preventing and / or treating symptoms associated with the deterioration of the condition of the application site of the composition of the present invention. In particular, the method of the present invention may be a method for preventing and / or treating symptoms associated with the deterioration of the condition of the skin.

[0066] The application methods of the composition of the present invention (for example, the target of application, the location of application, the timing of application, the duration of application, the number of applications, the amount applied, and other conditions related to application) are not particularly limited as long as the desired effect is obtained. The usage methods of the composition of the present invention can be appropriately set according to various conditions such as the type and content of the active ingredient, the type and content of other ingredients, the type and shape (dosage form) of the composition, the type of target of application, age, and health condition, and the purpose of use of the composition of the present invention.

[0067] The applicable subjects and application locations of the composition of the present invention are as described above. That is, the composition of the present invention can be applied to the subjects and application locations exemplified above, for example. The composition of the present invention can be applied to living organisms by appropriate methods such as coating, pasting, or spraying.

[0068] The application period of the composition of the present invention may be, for example, 1 hour or more, 6 hours or more, 1 day or more, 3 days or more, 1 week or more, 4 weeks or more, 2 months or more, 6 months or more, 12 months or more, 2 years or more, 5 years or more, or 10 years or more. The composition of the present invention may be applied, for example, daily or only during a specific period. The composition of the present invention may be applied, for example, continuously or intermittently. The composition of the present invention may be applied, for example, until the desired effect is obtained. The composition of the present invention may be applied, for example, once a day or divided into multiple times a day. Also, the composition of the present invention may be applied, for example, every day or once every few days. The application amount of the composition of the present invention at each application may be constant or not when converted to the application amount of the active ingredient. The single application amount of the composition of the present invention, when converted to the application amount of the active ingredient, is, for example, 0.1 μg / cm 2 or more, 0.5 μg / cm 2 or more, 1 μg / cm 2 or more, 5 μg / cm 2 or more, or 10 μg / cm 2 or more, and may be 500 mg / cm 2 or less, 100 mg / cm 2 or less, 50 mg / cm 2 or less, 10 mg / cm 2 or less, 5 mg / cm 2 or less, 1 mg / cm 2 or less, 0.5 mg / cm 2 or less, or 0.1 mg / cm 2 or less, and may also be a combination thereof. The single application amount of the composition of the present invention, when converted to the application amount of the active ingredient, specifically, for example, is 0.1 μg / cm 2 ~500 mg / cm 2 and may be so.

[0069] <3>Production method of PHC The following describes methods for producing PHC or PHS by fermentation using yeast, and methods for producing PHC by conversion of PHS. The product of the fermentation method (i.e., PHC and / or PHS) is also referred to as the "target substance." The yeast used in the fermentation method is also referred to as the "yeast of the present invention."

[0070] <3-1> The Yeast of the Invention The yeast of this invention is a yeast that has the ability to produce the target substance. The "ability to produce the target substance" is also called the "target substance production capacity."

[0071] In the present invention, "yeast having the ability to produce the target substance" refers to yeast that, when cultured in a culture medium, can produce the target substance and accumulate it in the culture medium and / or within the cells to a degree that allows for its recovery. The culture medium may be a culture medium that can be used in a fermentation method, and specifically, it may be a culture medium containing fatty acids. The yeast having the ability to produce the target substance may be yeast that can accumulate a larger amount of the target substance in the culture medium and / or within the cells than an unmodified strain. "Unmodified strain" may mean a control strain that has not been modified to confer or enhance the ability to produce the target substance. That is, examples of unmodified strains include wild strains and parent strains, such as Saccharomyces cerevisiae strain BY4742 (ATCC 201389; EUROSCARF Y10000), strain S288C (ATCC 26108), and strain NCYC 3608. Furthermore, the yeast having the ability to produce the target substance may be yeast that can accumulate an amount of the target substance in the culture medium preferably of 5 mg / L or more, more preferably of 10 mg / L or more.

[0072] The yeast is not particularly limited as long as it can be used in the fermentation method. The yeast may be budding yeast or fission yeast. The yeast may be haploid, diploid, or more polyploid.

[0073] Examples of yeasts include those belonging to the genus Saccharomyces, such as Saccharomyces cerevisiae; the genus Pichia (also known as the genus Wickerhamomyces), such as Pichia ciferrii, Pichia sydowiorum, and Pichia pastoris; the genus Candida, such as Candida utilis; the genus Hansenula, such as Hansenula polymorpha; and the genus Schizosaccharomyces, such as Schizosaccharomyces pombe. Some species of the genus Pichia have been reclassified into the genus Wickerhamomyces (Int J Syst Evol Microbiol. 2014 Mar;64(Pt 3):1057-61). Therefore, for example, Pichia ciferrii and Pichia sydowiorum are also called Wickerhamomyces ciferrii and Wickerhamomyces sydowiorum, respectively. In this invention, "Pichia" includes species that were formerly classified in the genus Pichia but have been reclassified into other genera such as Wickerhamomyces.

[0074] Examples of Saccharomyces cerevisiae include strain BY4742 (ATCC 201389; EUROSCARF Y10000), strain S288C (ATCC 26108), strain Y006 (FERM BP-11299), strain NCYC 3608, and their derivatives. Examples of Pichia ciferrii (Wickerhamomyces ciferrii) include strain NRRL Y-1031 (ATCC 14091), strain CS.PCΔPro2 (Schorsch et al., 2009, Curr Genet. 55, 381-9.), the strain disclosed in WO95 / 12683, and their derivatives. Specifically, examples of Pichia sydowiorum (Wickerhamomyces sydowiorum) include the NRRL Y-7130 strain (ATCC 58369) and its derivatives.

[0075] These strains are, for example, from the American Type Culture Collection (ATCC, Address: PO Box 1549, Manassas, VA 20108, United States of America) and the European Saccharomyces. Cerevisiae strains can be obtained from the Cerevisiae Archive for Functional Analysis (EUROSCARF, Address: Institute for Molecular Biosciences, Johann Wolfgang Goethe-University Frankfurt, Max-von-Laue Str. 9; Building N250, D-60438 Frankfurt, Germany), the National Collection of Yeast Cultures (NCYC, Address: Institute of Food Research, Norwich Research Park, Norwich, NR4 7UA, UK), or from the depositary institution corresponding to each deposited strain. For example, in the case of ATCC strains, each strain is assigned a registration number, which can be used to place an order (see http: / / www.atcc.org / ). The registration numbers for each strain are listed in the catalog of the American Type Culture Collection (ATCC).

[0076] The yeast of the present invention may inherently possess the ability to produce the target substance, or it may be modified to possess the ability to produce the target substance. Yeast possessing the ability to produce the target substance can be obtained by conferring the ability to produce the target substance to the above-mentioned yeast, or by enhancing the ability to produce the target substance of the above-mentioned yeast.

[0077] The following are specific examples of methods for conferring or enhancing the ability to produce the target substance. The modifications for conferring or enhancing the ability to produce the target substance may be used individually or in appropriate combinations. The modifications for constructing the yeast of the present invention may be carried out in any order.

[0078] The ability to produce the target substance can be conferred or enhanced by modifying the yeast so that the expression and / or activity of one or more proteins involved in the production of the target substance is increased or decreased. That is, the yeast of the present invention may be modified so that the expression and / or activity of one or more proteins involved in the production of the target substance is increased or decreased. "Protein" includes so-called peptides such as polypeptides. Proteins involved in the production of the target substance include enzymes that catalyze the synthesis of the target substance (hereinafter also referred to as "biosynthetic enzymes of the target substance"), enzymes that catalyze reactions that branch off from the biosynthetic pathway of the target substance and produce compounds other than the target substance (hereinafter also referred to as "biosynthetic enzymes of by-products"), enzymes that catalyze the degradation of the target substance (hereinafter also referred to as "degrading enzymes of the target substance"), and proteins that affect (for example increase or decrease) the activity of such enzymes.

[0079] The proteins whose expression and / or activity are increased or decreased can be appropriately selected depending on the type of target substance, the proteins involved in the production of the target substance, and the types and activities of proteins inherently present in the yeast of the present invention. For example, preferably, the expression and / or activity of one or more proteins selected from the biosynthetic enzymes of the target substance may be increased. Alternatively, preferably, the expression and / or activity of one or more proteins selected from the biosynthetic enzymes of by-products and the degrading enzymes of the target substance may be decreased.

[0080] The activity of a protein can be increased, for example, by increasing the expression of the gene encoding that protein. The expression and / or activity of a protein can be increased, for example, by increasing the copy number of the gene encoding that protein, or by modifying the regulatory sequence of the gene encoding that protein. The activity of a protein can be decreased, for example, by decreasing the expression of the gene encoding that protein, or by disrupting the gene encoding that protein. The expression and / or activity of a protein can be decreased, for example, by deletion of the gene encoding that protein. When increasing or decreasing the expression and / or activity of two or more proteins, the methods for increasing or decreasing the expression and / or activity of each protein can be selected independently. Gene expression can be controlled by "protein This is also called "protein expression" (i.e., the protein encoded by the gene). Methods for increasing or decreasing the expression and / or activity of such proteins are well known in the art.

[0081] Specifically, the proteins involved in the production of the target substance are LCB1, LCB2, TSC10, and SUR2. These include proteins encoded by the LAG1, LAC1, LIP1, SER1, SER2, SER3, YPC1, NEM1, SPO7, LCB4, LCB5, ELO3, CKA2, ORM2, and CHA1 genes.

[0082] The yeast of the present invention includes LCB1, LCB2, TSC10, SUR2, LAG1, LAC1, LIP1, SER1, SER2, SER3, The Ypc1 genes may be modified to increase the expression and / or activity of one or more proteins encoded by the YPC1 gene, and / or to decrease the expression and / or activity of one or more proteins encoded by the YPC1, NEM1, SPO7, LCB4, LCB5, ELO3, CKA2, ORM2, and CHA1 genes. The expression and / or activity of Ypc1p may be increased, for example, when producing PHS. The expression and / or activity of Ypc1p may be decreased, for example, when producing PHC.

[0083] Examples of these genes and proteins include those from yeasts such as S. cerevisiae and Pichia ciferrii.

[0084] The genes and proteins used in breeding the yeast of the present invention may, for example, have the base sequence and amino acid sequence of known genes and proteins, such as those exemplified above. Furthermore, the genes and proteins used in breeding L-amino acid-producing bacteria may, for example, be variants of known genes and proteins, such as those exemplified above, as long as their original function (e.g., activity or properties) is maintained.

[0085] <3-2> Method for manufacturing PHC The fermentation method can be carried out by culturing the yeast of the present invention in a culture medium. In the fermentation method, one target substance may be produced, or two or more target substances may be produced.

[0086] The culture medium used is not particularly limited, as long as it allows the yeast of the present invention to grow and produces the target substance. For example, a conventional culture medium used for yeast cultivation can be used. Examples of such media include SD medium, SG medium, SDTE medium, and YPD medium. The culture medium may contain, as needed, a carbon source, a nitrogen source, a phosphorus source, a sulfur source, and other components selected from various organic and inorganic components. The types and concentrations of the culture medium components may be appropriately set according to various conditions such as the type of yeast used and the type of target substance to be produced.

[0087] The culture medium may contain fatty acids. The use of fatty acids may increase the production of the target substance. That is, in the presence of fatty acids, the production of the target substance by yeast may increase compared to the absence of fatty acids. Increases in the production of the target substance include an increase in the amount of the target substance produced, an increase in the production rate of the target substance, and an increase in the yield of the target substance. In addition, the use of fatty acids may allow for adjustment of the composition (e.g., length) of the alkyl chain of the target substance. Such adjustments include adjusting the production of the target substance containing a specific alkyl chain, or adjusting the ratio of the production amount of the target substance containing a specific alkyl chain to the total amount of all products. Such a ratio is also called the "production ratio". A specific alkyl chain may be an alkyl chain of a specific length. "Total amount of all products" may mean, for example, the total amount of two or more PHS species (e.g., all PHS species produced) or two or more PHC species (e.g., all PHC species produced).

[0088] In other words, specifically, the use of fatty acids, depending on the type of fatty acid, results in the formation of certain alkyl groups. The production of the target substance containing the chain may be increased. For example, by using a fatty acid with n carbon atoms, The production of the target substance containing an alkyl chain of n+2 may increase. In other words, if the number of carbon atoms in the fatty acid is n, the target substance contains a PHS or PHC species containing an alkyl chain of n+2 carbon atoms. It is acceptable for the production of the PHS species or PHC species to increase due to the presence of fatty acids. "The target substance contains a PHS species or PHC species" means that at least the PHS species or PHC species are produced as the target substance, and the target substance consists of the PHS species or PHC species, or contains the PHS species. This may include cases where the mixture consists of a PHC species.

[0089] Furthermore, specifically, the use of fatty acids may increase the ratio of the production amount of a target substance containing a specific alkyl chain to the total amount of products, depending on the type of fatty acid. For example, a fatty acid with n carbon atoms The use of fatty acids may increase the ratio of the production of the target substance containing an alkyl chain with n+2 carbon atoms to the total production amount. In other words, if the number of carbon atoms in the fatty acid is n, the target substance is The product may contain PHS species or PHC species that include an alkyl chain with n+2 carbon atoms, and the ratio of the production of said PHS species or PHC species to the total amount of product may increase due to the presence of fatty acids.

[0090] The length and degree of unsaturation of fatty acids may be variable. The length of a fatty acid may be, for example, C12-C24 (C12, C14, C16, C18, C20, C22, C24, etc.). The length of a fatty acid may, for example, be C14, C16, or C18. The length of a fatty acid can be interpreted as the number of carbon atoms (i.e., the number of carbon atoms) in the fatty acid. Fatty acids may be saturated or unsaturated. Fatty acids may have one or more unsaturated double bonds. Specifically, examples of fatty acids include lauric acid (12:0), myristic acid (14:0), palmitic acid (16:0), stearic acid (18:0), arachidic acid (20:0), behenic acid (22:0), lignoceric acid (24:0), myristoleic acid (14:1), palmitoleic acid (16:1), oleic acid (18:1), linoleic acid (18:2), and linolenic acid (18:3). In particular, examples of fatty acids include myristic acid (14:0), palmitic acid (16:0), and stearic acid (18:0). Even more particularly, examples of fatty acids include myristic acid (14:0). The use of myristic acid (14:0) may increase the production volume or production ratio of C16 PHS or PHC, such as C16:0 PHS or PHC. The use of palmitic acid (16:0) may increase the production volume or production ratio of C18 PHS or PHC, for example, C18:0 PHS or PHC. The use of stearic acid (18:0) may increase the production volume or production ratio of C20 PHS or PHC, for example, C20:0 PHS or PHC. One type of fatty acid may be used as the fatty acid, or two or more fatty acids may be used in combination.

[0091] The fatty acid may be in its free form, its salt, or a mixture thereof. That is, in this invention, unless otherwise specified, the term "fatty acid" may mean a free fatty acid, its salt, or a mixture thereof. Examples of salts include ammonium salts, sodium salts, and potassium salts. As the precursor salt, one type of salt may be used, or two or more salts may be used in combination.

[0092] Fatty acids may be present in the culture medium for the entire duration of the culture, or for only a portion of the culture period. In other words, "cultivating yeast in a culture medium containing fatty acids" does not necessarily mean that fatty acids are present in the culture medium for the entire duration of the culture. For example, fatty acids may or may not be present in the culture medium from the start of the culture. If fatty acids are not present in the culture medium at the start of the culture, fatty acids are supplied to the culture medium after the start of the culture. The timing of the supply can be appropriately set according to various conditions such as the length of the culture period. For example, fatty acids may be supplied to the culture medium after the yeast of the present invention has grown sufficiently. In any case, fatty acids may be supplied to the culture medium in addition as needed. The means of supplying fatty acids to the culture medium are not particularly limited. For example, fatty acids can be supplied to the culture medium by supplying a fed-batch medium containing fatty acids to the culture medium. The concentration of fatty acids in the culture medium is not particularly limited as long as the target substance can be produced. For example, the concentration of fatty acids in the culture medium may be 0.1 g / L or more, 1 g / L or more, 2 g / L or more, 5 g / L or more, or 10 g / L or more, or 200 g / L or less, 100 g / L or less, 50 g / L or less, or 20 g / L or less, or a combination thereof. The concentration of fatty acids in the culture medium may be, for example, 0.1 g / L to 200 g / L, 1 g / L to 100 g / L, or 5 g / L to 50 g / L. Fatty acids may be contained in the culture medium at concentrations within the above example ranges throughout the entire period of culture, or not. For example, fatty acids may be contained in the culture medium at concentrations within the above example ranges at the start of culture, or they may be supplied to the culture medium after the start of culture so that the concentration becomes within the above example ranges.

[0093] The culture medium may contain additives that enable association with, binding to, solubilization of, and / or capture of the target substance (WO2017 / 033463). The use of additives allows for: The production of the target substance may increase. That is, in the presence of the additive, the amount of the target substance produced by the yeast of the present invention may increase compared to the absence of the additive. Specifically, the use of the additive may increase the production of the target substance in the culture medium. The production of the target substance in the culture medium is also called "excretion of the target substance." "Association with the target substance, binding with the target substance, solubilization of the target substance, and / or capture of the target substance" may specifically mean increasing the solubility of the target substance in the culture medium. Examples of additives include cyclodextrins and zeolites. The number of glucose residues constituting the cyclodextrin is not particularly limited. The number of glucose residues constituting the cyclodextrin may be, for example, 5, 6, 7, or 8. That is, examples of cyclodextrins include cyclodextrins consisting of 5 glucose residues, α-cyclodextrins, β-cyclodextrins, γ-cyclodextrins, and their derivatives. Examples of cyclodextrin derivatives include cyclodextrins to which one or more functional groups have been introduced. The type, number, and amount of functional groups, as well as the position in which the functional groups are introduced, are not particularly limited, as long as the derivative associates with the target substance, binds to the target substance, solubilizes the target substance, and / or captures the target substance. Functional groups may be introduced, for example, at the C2 hydroxyl group, the C3 hydroxyl group, the C6 hydroxyl group, or a combination thereof, thereby increasing the solubility of the cyclodextrin itself. Examples of functional groups include alkyl groups and hydroxyalkyl groups. Both alkyl groups and hydroxyalkyl groups may have a linear alkyl chain or a branched alkyl chain. Both alkyl groups and hydroxyalkyl groups may have, for example, 1, 2, 3, 4, or 5 carbon atoms. Specific examples of alkyl groups include methyl, ethyl, propyl, butyl, pentyl, isopropyl, and isobutyl groups. Specific examples of hydroxyalkyl groups include hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, hydroxypentyl, hydroxyisopropyl, and hydroxyisobutyl groups.Examples of cyclodextrin derivatives include methyl-α-cyclodextrin, methyl-β-cyclodextrin, hydroxypropyl-α-cyclodextrins such as 2-hydroxypropyl-α-cyclodextrin, and hydroxypropyl-β-cyclodextrins such as 2-hydroxypropyl-β-cyclodextrin. The type of zeolite is not particularly limited. As for additives, one additive may be used, or two or more additives may be used in combination.

[0094] The additive may be present in the culture medium for the entire duration of the culture, or for only a portion of the culture period. In other words, "cultivating yeast in a culture medium containing an additive" does not necessarily mean that the additive is present in the culture medium for the entire duration of the culture. For example, the additive may or may not be present in the culture medium from the start of the culture. If the culture medium does not contain the additive at the start of the culture, the additive is supplied to the culture medium after the start of the culture. The timing of the supply can be appropriately set according to various conditions such as the length of the culture period. For example, the additive may be supplied to the culture medium after the yeast of the present invention has grown sufficiently. In any case, the additive may be supplied to the culture medium in addition as needed. The means of supplying the additive to the culture medium are not particularly limited. For example, the additive can be supplied to the culture medium by feeding a fed-batch medium containing the additive into the culture medium. The concentration of the additive in the culture medium is not particularly limited as long as the target substance can be produced. The concentration of the additive in the culture medium may be, for example, 0.1 g / L or more, 1 g / L or more, 2 g / L or more, 5 g / L or more, or 10 g / L or more, or 200 g / L or less, 100 g / L or less, 50 g / L or less, or 20 g / L or less, or a combination thereof. The concentration of the additive in the culture medium may be, for example, 0.1 g / L to 200 g / L, 1 g / L to 100 g / L, or 5 g / L to 50 g / L. The additive may be contained in the culture medium at concentrations within the above example ranges throughout the entire period of culture, or not. For example, the additive may be contained in the culture medium at concentrations within the above example ranges at the start of culture, or it may be supplied to the culture medium after the start of culture to reach concentrations within the above example ranges.

[0095] Specifically, examples of carbon sources include sugars such as glucose, fructose, sucrose, lactose, galactose, xylose, arabinose, molasses, starch hydrolysates, and biomass hydrolysates; organic acids such as acetic acid, fumaric acid, citric acid, and succinic acid; alcohols such as glycerol, crude glycerol, and ethanol; and fatty acids. In other words, the fatty acids mentioned above may also be used as carbon sources. The fatty acids mentioned above may be used as the sole carbon source, or they may not. However, usually, at least one carbon source other than the fatty acids mentioned above may be used. One carbon source may be used, or two or more carbon sources may be used in combination.

[0096] Specific examples of nitrogen sources include ammonium salts such as ammonium sulfate, ammonium chloride, and ammonium phosphate; peptone; organic nitrogen sources such as yeast extract, meat extract, and soy protein hydrolysate; ammonia; and urea. Ammonia gas or ammonia water used for pH adjustment may also be used as a nitrogen source. One nitrogen source may be used, or two or more nitrogen sources may be used in combination.

[0097] Examples of phosphate sources include phosphates such as potassium dihydrogen phosphate and dipotassium hydrogen phosphate, and phosphate polymers such as pyrophosphate. One phosphate source may be used, or two or more phosphate sources may be used in combination.

[0098] Examples of sulfur sources include inorganic sulfur compounds such as sulfates, thiosulfates, and sulfites, and sulfur-containing amino acids such as cysteine, cystine, and glutathione. A single sulfur source may be used, or a combination of two or more sulfur sources may be used.

[0099] Other various organic and inorganic components include, specifically, inorganic salts such as sodium chloride and potassium chloride; trace metals such as iron, manganese, magnesium, and calcium; and vitamins such as vitamin B1, vitamin B2, vitamin B6, nicotinic acid, nicotinamide, and vitamin B12. Examples of organic components include methyl compounds; amino acids; nucleic acids; and peptones, casamino acids, yeast extracts, and hydrolyzed soy protein products containing these compounds. Other organic and inorganic components may be used individually, or in combination of two or more components.

[0100] Furthermore, when using nutrient-dependent mutant strains that require amino acids, nucleic acids, etc. for growth, it is preferable to supplement the culture medium with the necessary nutrients.

[0101] The culture conditions are not particularly limited, as long as the yeast of the present invention can grow and the target substance can be produced. Culture can be carried out, for example, under the usual conditions used for yeast culture. The culture conditions may be set appropriately depending on various conditions such as the type of yeast used and the type of target substance to be produced.

[0102] Culturing can be carried out using liquid medium under aerobic, microaerobic, or anaerobic conditions. The culture can preferably be carried out under aerobic conditions. "Aerobic conditions" means that the dissolved oxygen concentration in the liquid medium is 0.33 ppm or higher, preferably 1.5 ppm or higher. Under aerobic conditions, the oxygen concentration may be controlled to, for example, 5-50%, preferably 10-20%, of the saturated oxygen concentration. Aerobic culture is specifically carried out by aeration or shaking. This is possible. "Microaerophilic conditions" may be conditions in which oxygen is supplied to the culture system, but the dissolved oxygen concentration in the liquid medium is less than 0.33 ppm. "Anaerobic conditions" may be conditions in which oxygen is not supplied to the culture system. The culture temperature may be, for example, 25 to 35°C, preferably 27 to 33°C, and more preferably 28 to 32°C. The pH of the medium may be, for example, pH 3 to 10, preferably pH 4 to 8. During cultivation, the pH of the medium can be adjusted as needed. pH adjustment is Inorganic or organic acidic or alkaline substances, such as ammonia gas, can be used. The culture period may be, for example, 10 to 200 hours, or 15 to 120 hours. Culture conditions may be constant throughout the entire culture period, or they may be changed during the culture. Culture can be carried out by batch culture, fed-batch culture, continuous culture, or a combination thereof. Culture may be carried out in two stages: seed culture and main culture. In such cases, the culture conditions for seed culture and main culture may or may not be the same. For example, both seed culture and main culture may be carried out by batch culture. Alternatively, for example, seed culture may be carried out by batch culture, and main culture may be carried out by fed-batch culture or continuous culture.

[0103] By culturing the yeast of the present invention under these conditions, the target substance accumulates in the culture medium and / or within the yeast cells.

[0104] The formation of the target substance can be confirmed by known methods used for the detection or identification of compounds. Examples of such methods include HPLC, UPLC, LC / MS, GC / MS, and NMR. These methods can be used individually or in appropriate combinations.

[0105] The generated target substance can be recovered as appropriate. Recovery of the generated target substance can be carried out by known methods used for the separation and purification of compounds. Examples of such methods include ion exchange resin methods, membrane treatment methods, precipitation methods, and crystallization methods. These methods can be used individually or in appropriate combinations. If the target substance accumulates within the bacterial cells, for example, the bacterial cells can be disrupted by ultrasound, and then the target substance can be recovered from the supernatant obtained by removing the bacterial cells by centrifugation. The recovered target substance may be a free form, its salt, or a mixture thereof.

[0106] Furthermore, if the target substance precipitates in the culture medium, it can be recovered by centrifugation or filtration. Alternatively, the target substance precipitated in the culture medium may be isolated together with the target substance dissolved in the culture medium after crystallization.

[0107] Furthermore, the recovered target substance may contain additional components other than the target substance, such as yeast cells, culture medium components, water, and metabolic by-products of yeast. The purity of the recovered target substance may be, for example, 30% (w / w) or higher, 50% (w / w) or higher, 70% (w / w) or higher, 80% (w / w) or higher, 90% (w / w) or higher, or 95%. It may be (w / w) or more.

[0108] When PHS is produced by culturing the yeast of the present invention, the produced PHS is converted to the corresponding PHC. This is possible. In other words, the present invention provides a method for producing PHC, comprising producing PHS by fermentation and converting the PHS into PHC.

[0109] The PHS produced by culturing the yeast of the present invention can be used as is, or concentrated as needed. After undergoing appropriate processing such as dilution, drying, dissolution, fractionation, extraction, and purification, it is used for conversion to PHC. This is possible. In other words, for PHS, for example, even if a product purified to a desired degree is used. Materials containing PHS may also be used. The materials containing PHS are not particularly limited as long as the conversion from PHS to PHC is progressing. Specific examples of materials containing PHS include PHS-containing culture media, supernatants separated from culture media, and processed products thereof (e.g., concentrates (concentrates, etc.) or dried products).

[0110] There are no particular restrictions on how PHS can be converted to PHC.

[0111] PHS can be converted to PHC, for example, by a chemical reaction with a fatty acid (J. Biol. Chem. July 2002 277 (29): 25847-5). The chemical reaction can be carried out under typical conditions for condensing an amine and a carboxylic acid to form an amide bond. Specifically, the chemical reaction can be carried out using a condensing agent. Examples of condensing agents include carbodiimides such as water-soluble carbodiimides (WSCs). A specific example of a WSC is 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride. The fatty acid is not particularly limited as long as it provides the acyl chain of the PHC to be produced. That is, examples of fatty acids include those corresponding to the acyl chains of the PHCs exemplified above. Specifically, examples of fatty acids include myristic acid (14:0), palmitic acid (16:0), stearic acid (18:0), arachidic acid (20:0), behenic acid (22:0), lignoceric acid (24:0), cerotic acid (26:0), myristoleic acid (14:1), palmitoleic acid (16:1), oleic acid (18:1), linoleic acid (18:2), and linolenic acid (18:3). Stearic acid (18:0) is a particularly noteworthy fatty acid. One type of PHS may be used as the PHS, or two or more types of PHS may be used in combination. One type of fatty acid may be used as the fatty acid, or two or more types of fatty acids may be used in combination. A mixture of two or more PHC types may be produced by using two or more PHS types and / or two or more fatty acids in combination.

[0112] Confirmation of PHC generation and recovery of PHC can be carried out in the same manner as by fermentation. The percentage may be, for example, 30% (w / w) or more, 50% (w / w) or more, 70% (w / w) or more, 80% (w / w) or more, 90% (w / w) or more, or 95% (w / w) or more.

[0113] The PHC produced as described above can be used as is, or concentrated, diluted, and dried as needed. After being subjected to appropriate processing such as dissolution, fractionation, extraction, and purification, it can be used as an active ingredient. That is, as an active ingredient, for example, a material purified to a desired degree may be used, or a material containing PHC may be used. The material containing PHC is not particularly limited as long as it is permissible according to the intended use of the composition of the present invention. Specifically, examples of materials containing PHC include PHC-containing culture medium or conversion reaction mixture, supernatant separated from culture medium or conversion reaction mixture, and processed products thereof (e.g., concentrates (concentrates, etc.) or dried products).

[0114] <4> Use of active ingredients Furthermore, the present invention discloses the use of the active ingredient in the applications exemplified above. Specifically, the present invention discloses, for example, the use of the active ingredient to improve the condition of the application site (for example, the application site of the composition of the present invention), and the use of the active ingredient in the manufacture of a composition (for example, a cosmetic or pharmaceutical composition) for improving the condition of the application site. Furthermore, the present invention discloses, for example, the use of the active ingredient to improve the properties of a composition (for example, a cosmetic or pharmaceutical composition).

[0115] Furthermore, the present invention discloses active ingredients for use in the applications exemplified above. Specifically, the present invention discloses, for example, active ingredients for use in improving the condition of an application site (for example, an application site of the composition of the present invention), and active ingredients for use in the manufacture of a composition (for example, a cosmetic or pharmaceutical composition) for improving the condition of an application site. Furthermore, the present invention discloses, for example, active ingredients for use in improving the properties of a composition (for example, a cosmetic or pharmaceutical composition). ru. [Examples]

[0116] The present invention will be described more specifically below with reference to non-limiting embodiments.

[0117] <1> Preparation of PHC The C18:0 PHS was obtained as a commercially available product from Tokyo Chemical Industries. The C16:0 PHS and C20:0 PHS were also obtained from Tokyo Chemical Industries. These PHS were obtained using the contract manufacturing service of Seikogyo. Each of these PHS was condensed with a fatty acid, specifically palmitic acid (16:0), stearic acid (18:0), or behenic acid (22:0), using the condensing agent WSCI-HCl (1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride; Watanabe Chemical Industry), to prepare the corresponding PHC species. In the examples, Cn:m PHS The PHC of Cx:y fatty acids is also called "Cx:y-PHSn:m".

[0118] <2> Analysis of the properties of PHC-containing compositions (1) <2-1> Preparation of the composition The amounts of PHC, cholesterol, and lauric acid shown in Table 1 were mixed. 95 After heating to a certain temperature to melt the material, it was cooled to room temperature, and each sample was prepared.

[0119] <2-2> Measurement of Melting Point Each sample was heated to 70°C and then further heated at a rate of 1°C / 5 minutes. The temperature at which the sample completely melted was defined as the melting point of the sample.

[0120] <2-3> Measurement of lamellar structure formation ability Each sample was coated onto a microscope slide, and a coverslip was placed on top. The slide was placed on a 120°C hot plate to melt the sample, and then allowed to cool. The thesis cross images were observed using a polarizing microscope. Based on the observation of the Malthesis cross images, the lamellar structure-forming ability of the samples was evaluated and classified on a score from 1 to 5 as follows. Score 5: Maltese crosses are observed throughout the entire field of view. Score 4: The Maltese cross image is observed across almost the entire field of view. Score 3: A Maltese cross image is observed in part of the field of view. Score 2: A faint image of the Maltese Cross is observed. Score 1: No image of the Maltese Cross is observed.

[0121] <2-4>Result The results are shown in Table 1. Samples containing combinations of three PHC species with different alkyl chain lengths Samples 1 and 2 have lower melting points and higher melting points than samples 3-5 which contain a single type of PHC species. The samples demonstrated lamellar structure formation ability. Furthermore, samples 1 and 2 showed higher lamellar structure formation ability than sample 6, which contained a combination of three PHC species with different acyl chain lengths. In other words, it was suggested that using multiple PHC species with different alkyl chain lengths in combination improves the properties of compositions containing them compared to using only a single PHC species or using multiple PHC species with different acyl chain lengths in combination.

[0122] [Table 1]

[0123] <3> Analysis of the properties of PHC-containing compositions (2) <3-1> Preparation of Composition PHCs in the proportions shown in Table 2 were mixed in a mortar to prepare each sample.

[0124] <3-2> Measurement of Melting Point Using a differential scanning calorimeter DSC7000X (Hitachi High-Tech Science), approximately 2 mg of each sample was heated in a nitrogen atmosphere from 25°C at a heating rate of 5°C / min. The baseline and the inflection point of the endothermic peak were measured. The temperature at the intersection of the lines was defined as the melting point.

[0125] <3-3>Results The results are shown in Table 2. Combinations of two or more PHC species with different alkyl chain lengths were selected. Samples 7-10 contain a single PHC species, as do sample 11 and 1. 2. Samples 12 and 13, samples 11 and 13, and samples 11-13 showed lower melting points. In other words, by using multiple PHC species with different alkyl chain lengths in combination again... Furthermore, compared to using only a single type of PHC, the properties of compositions containing them are It was suggested that it would improve.

[0126] [Table 2]

[0127] <4> Evaluation of skin barrier function restoration by PHC-containing compositions based on transepidermal water loss (TEWL) using isolated cadaveric skin. <4-1> Preparation of PHC mixture A PHC mixture with the composition shown in Table 3 was obtained. This PHC mixture or each PHC species contained therein For example, it can be reproducibly prepared by the method exemplified above.

[0128] [Table 3]

[0129] <4-2> Preparation of Composition The PHC mixture obtained in <4-1> or Ceramide III (Evonik) was mixed with fatty acids and cholesterol in the amounts shown in Table 4 to prepare lipid mixtures. Ceramide III (Evonik) consists of a single PHC species, C18:0-PHS18:0. Each lipid mixture was dissolved in chloroform / methanol. Each sample was prepared by dissolving the substance in chloroform:methanol (2:1) at a concentration of 1% by weight.

[0130] [Table 4]

[0131] <4-3> Measurement of transepidermal water loss (TEWL) <4-3-1> Skin Pretreatment The isolated cadaveric skin (KAC, frozen, full-thickness) is thawed at room temperature and immersed in chloroform / methanol (chloroform:methanol = 2:1, the same as in the following procedure) for 5 minutes. It was left to stand. The skin was removed from the chloroform / methanol solution and cut off the skin with a scalpel or scissors. Adipose tissue was removed. The skin was immersed in a 0.5% SLS (sodium lauryl sulfate) aqueous solution and shaken for 30 minutes. The skin was removed from the SLS aqueous solution and immersed in distilled water and shaken for 30 minutes. The skin was removed from the distilled water, immersed in fresh distilled water, and immediately removed. Moisture from the skin surface was removed with paper towels such as Kimwipes. The skin was immersed in chloroform / methanol and left to stand for 60 minutes. The skin was removed from the chloroform / methanol solution and placed on a soft piece of paper moistened with distilled water. The skin was placed with the dermis side down (i.e., with the dermis side facing the paper) and left to stand for 60 minutes to air dry.

[0132] <4-3-2> Determination of Application Amount An experiment was conducted to determine the appropriate sample dosage using the pre-treated skin obtained in <4-3-1>. 50 μl, 40 μl, or 30 μl of each sample was dropped onto the stratum corneum side of the skin and air-dried. Aluminum foil was placed on an 80°C block heater, and the skin was placed on top of the foil with the stratum corneum side down. The sample was heated for 2 minutes to melt and penetrate the skin. During heating, a container of ice water was placed on the upper side of the skin (i.e., the dermis side). After heating, the skin and aluminum foil were visually inspected, and the maximum dosage for each sample that left almost no sample on either the skin or the aluminum foil was determined.

[0133] <4-3-3> Measurement of Transepidermal Water Loss (TEWL) A 24-well plate (Corning; product number 353047) was placed on the stratum corneum side of the pre-treated skin obtained in <4-3-1> and secured with a rubber band. A weight was then placed on top and left to stand for 2 hours. Then, multiple circular patterns with a diameter of approximately 1.6 cm were formed in the stratum corneum. The 24-well plate was removed from the skin. Each sample, with the maximum application amount determined in <4-3-2>, was formed on the stratum corneum side of the skin. The sample was dropped onto a circular pattern and air-dried. Aluminum foil was placed on an 80°C block heater, and the skin was placed on top with the stratum corneum side down and heated for 2 minutes to melt the sample and immerse it in the skin. The skin was allowed to permeate. During heating, a container of ice water was placed on the upper side of the skin (i.e., the dermis side). After heating, the skin was placed dermis-side down (i.e., with the dermis facing the paper) on a paper moistened with distilled water, and left to stand in a sealed container at 30°C for at least 16 hours. The skin was removed from the sealed container and placed on a paper moistened with distilled water on a 26°C block heater placed in a 20°C room, and left to stand for 90 minutes. Transepidermal water loss (TEWL) of the skin was measured using Tewameter TM300 (Courage+Khazaka) and MPA580 (Courage+Khazaka). Skin TEWL was calculated as a relative value with the value of the untreated portion of the skin set to 100%. Here, "untreated portion" refers to the part of the skin to which the sample was not applied.

[0134] The results are shown in Figure 1. The TEWL in the portion treated with sample 14 containing the PHC mixture remains untreated. Although significantly lower than the TEWL of the untreated portion, the TEWL of the portion treated with sample 15 containing ceramide III (i.e., C18:0-PHS18:0) did not differ significantly from that of the untreated portion. In other words, the application of sample 14 significantly improved (restored) the skin barrier function. That is, again, it was suggested that the combined use of multiple PHC species with different alkyl chain lengths improves the properties of compositions containing them compared to the use of a single PHC species alone. [Industrial applicability]

[0135] According to the present invention, compositions such as cosmetic compositions can be provided.

Claims

1. A composition containing the following phytoceramides (A), (B), and (C): (A) A phytoceramide having a C16 phytosphingosine moiety in an amount of X weight% relative to the total amount of phytoceramides (A), (B), and (C); (B) A phytoceramide having a C18 phytosphingosine moiety in an amount of Y by weight % relative to the total amount of phytoceramides (A), (B), and (C); (C) A phytoceramide having a C20 phytosphingosine moiety in an amount of Z weight% relative to the total amount of phytoceramides (A), (B), and (C); X, Y, and Z are all greater than 0, and X, Y, and Z are all less than 100.

2. The composition according to claim 1, which is a cosmetic composition or a pharmaceutical composition.

3. Furthermore, the composition according to claim 1 or 2, further containing an ingredient for cosmetic or pharmaceutical use.

4. The composition according to claim 3, wherein the cosmetic or pharmaceutical ingredient is selected from the group consisting of excipients, binders, disintegrants, lubricants, stabilizers, diluents, surfactants, pH adjusters, vitamins, minerals, fragrances, pigments, preservatives, terpenoids, lipids, fatty acids, alcohols, water, antioxidants, anti-inflammatory agents, moisturizers, anti-aging agents, anti-cellulite agents, skin whitening agents, skin tanning agents, UV filters, and combinations thereof.

5. The composition according to claim 4, wherein at least a terpenoid, a lipid, a fatty acid, or a combination thereof is selected.

6. The composition according to claim 4 or 5, wherein the terpenoid is selected from the group consisting of cholesterol, cholesterol sulfate, squalene, pristane, and combinations thereof.

7. The composition according to any one of claims 4 to 6, wherein the lipid is selected from the group consisting of triolein, phosphatidylethanolamine, and combinations thereof.

8. The composition according to any one of claims 4 to 7, wherein the fatty acid is selected from the group consisting of lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid, oleic acid, linolenic acid, and combinations thereof.

9. The composition according to any one of claims 1 to 8, wherein X is 1 to 85, Y is 10 to 90, and Z is 2 to 40.

10. The composition according to any one of claims 1 to 9, wherein X is 65 to 90, Y is 5 to 30, and Z is 1 to 20.

11. The phytoceramide (A) is a phytoceramide having a C16:0 phytosphingosine moiety. Including Do, The phytoceramide (B) is a phytoceramide having a C18:0 phytosphingosine moiety. It includes and The phytoceramide (C) is a phytoceramide having a C20:0 phytosphingosine moiety. A composition according to any one of claims 1 to 10, comprising D.

12. The phytoceramide (A) has an acyl chain of a length selected from C14 to C26, The phytoceramide (B) has an acyl chain of a length selected from C14 to C26, and The composition according to any one of claims 1 to 11, wherein the phytoceramide (C) has an acyl chain of a length selected from C14 to C26.

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