JP7187781B2 - moisturizer - Google Patents

Description

The present invention relates to moisturizers and the like.

The stratum corneum has, as stratum corneum functions, a moisturizing ability to retain moisture inside and a barrier ability to suppress invasion of foreign substances (eg, foreign substances, viruses and bacteria) from the outside. Sebum, intercellular lipids, natural moisturizing factors and the like are known as substances that act on such functions of the stratum corneum.

Profilaggrin stored in the granular layer of skin tissue is degraded to filaggrin through dephosphorylation during keratinization. Filaggrin moves from the stratum granulosum to the stratum corneum, binds and aggregates with keratin fibers, and forms a keratin pattern. After that, filaggrin is decomposed into low-molecular-weight substances such as urocanic acid and becomes a natural moisturizing factor involved in water retention and ultraviolet absorption. Therefore, it is considered that the more the production of filaggrin is promoted, the more the amount of natural moisturizing factor, which is a decomposition product of filaggrin, increases, and the moisturizing ability of the skin improves.

Invasion of foreign substances from the outside world is suppressed by intercellular lipids and the like present between corneocytes that constitute the stratum corneum. Therefore, if the number, size, etc. of corneocytes are insufficient, gaps are formed in the stratum corneum, and the barrier function is lowered.

A peripheral zone, which is an extremely tough and huge insoluble structure lining the cell membrane of the corneocyte, exists around the corneocyte, which contains aggregated structures of keratin such as keratin 10 and filaggrin. The main structural elements of the marginal zone are proteins such as involucrin produced in spinous cells and loricrin produced in granule cells, which are cross-linked by enzymes such as transglutaminase (TG)1. Therefore, filaggrin, involucrin, transglutaminase 1, keratin 10, and loricrin strongly affect the formation of corneocytes, and are considered to be related to skin barrier function.

It has been reported that the expression of filaggrin, involucrin, transglutaminase 1, keratin 10, and loricrin is promoted by specific peptides such as glycylproline (Patent Document 1).

WO2014/175001

However, certain peptides such as glycylproline are weak in promoting the expression of moisturizing-related proteins such as filaggrin, and are considered to be insufficient in improving the moisturizing ability to retain moisture inside the stratum corneum. Such specific peptides are also weak in promoting the expression of proteins associated with the skin's barrier function, and are considered to be insufficient in improving the barrier function that suppresses invasion of foreign matter. That is, such specific peptides are considered insufficient for improving keratin function.

An object of the present invention is to provide a moisturizing agent capable of improving the moisturizing ability of retaining moisture inside the stratum corneum.

As a result of intensive studies, the present inventors found that a specific acyl neutral amino acid ester can improve the production of proteins related to the improvement of the moisturizing ability to retain moisture inside the stratum corneum, and is therefore promising as a moisturizing agent. I found The present inventors also found that a specific acyl neutral amino acid ester is useful as a high value-added moisturizer that can also improve the barrier ability to suppress the invasion of foreign substances, and completed the present invention. .

〔式中、
Ａは、炭素原子数１～１７の直鎖もしくは分岐鎖の炭化水素基であり、
Ｂは、炭素原子数２～１８の直鎖もしくは分岐鎖の炭化水素基であり、
Ｘは、水素原子、または炭素原子数１～５の直鎖もしくは分岐鎖の炭化水素基である。〕で表される化合物を含む、保湿剤。
〔２〕前記式（１）で表される化合物が、下記式（２）～（６）：

〔式中、
ＡおよびＢはそれぞれ、前記式（１）のものと同じである。〕のいずれか一つにより表される化合物である、〔１〕の保湿剤。
〔３〕Ａが、炭素原子数１～１１の直鎖もしくは分岐鎖の炭化水素基である、〔１〕または〔２〕の保湿剤。
〔４〕Ｂが、炭素原子数２～８の直鎖もしくは分岐鎖の炭化水素基である、〔１〕～〔３〕のいずれかの保湿剤。
〔５〕Ｂが、炭素原子数２もしくは３の直鎖もしくは分岐鎖の炭化水素基である、〔４〕の保湿剤。
〔６〕Ａが、炭素原子数７～１７の直鎖もしくは分岐鎖の炭化水素基である、〔１〕～〔５〕のいずれかの保湿剤。
〔７〕Ａ、ＢおよびＸにおける炭化水素基がアルキルである、〔１〕～〔６〕のいずれかの保湿剤。
〔８〕角層機能改善作用を有する、〔１〕～〔７〕のいずれかの保湿剤。
〔９〕プロフィラグリン、フィラグリン、インボルクリン、トランスグルタミナーゼ１、ケラチン１０、およびロリクリンからなる群より選ばれる１または２以上のタンパク質の産生促進作用を有する、〔１〕～〔８〕のいずれかの保湿剤。
〔１０〕前記化合物を０．０１～１０００ｍＭの濃度で含む組成物である、〔１〕～〔９〕のいずれかの保湿剤。
〔１１〕保湿剤が皮膚外用剤である、〔１〕～〔１０〕のいずれかの保湿剤。
〔１２〕保湿剤が化粧料である、〔１〕～〔１１〕のいずれかの保湿剤。
〔１３〕化粧料がリーブオン化粧料である、〔１２〕の保湿剤。 That is, the present invention is as follows.
[1] Formula (1) below:

[In the formula,
A is a linear or branched hydrocarbon group having 1 to 17 carbon atoms,
B is a linear or branched hydrocarbon group having 2 to 18 carbon atoms,
X is a hydrogen atom or a linear or branched hydrocarbon group having 1 to 5 carbon atoms. ] A moisturizing agent containing a compound represented by
[2] The compound represented by the above formula (1) is represented by the following formulas (2) to (6):

[In the formula,
A and B are each the same as in formula (1) above. ], the moisturizing agent of [1], which is a compound represented by any one of [1].
[3] The moisturizer of [1] or [2], wherein A is a linear or branched hydrocarbon group having 1 to 11 carbon atoms.
[4] The moisturizer of any one of [1] to [3], wherein B is a straight or branched hydrocarbon group having 2 to 8 carbon atoms.
[5] The moisturizer of [4], wherein B is a linear or branched hydrocarbon group having 2 or 3 carbon atoms.
[6] The moisturizing agent of any one of [1] to [5], wherein A is a straight or branched hydrocarbon group having 7 to 17 carbon atoms.
[7] The moisturizer of any one of [1] to [6], wherein the hydrocarbon groups in A, B and X are alkyl.
[8] The moisturizing agent of any one of [1] to [7], which has an effect of improving stratum corneum function.
[9] The moisturizing agent of any one of [1] to [8], which has the action of promoting the production of one or more proteins selected from the group consisting of profilaggrin, filaggrin, involucrin, transglutaminase 1, keratin 10, and loricrin. agent.
[10] The moisturizing agent of any one of [1] to [9], which is a composition containing the compound at a concentration of 0.01 to 1000 mM.
[11] The moisturizing agent of any one of [1] to [10], which is an external preparation for skin.
[12] The moisturizer of any one of [1] to [11], which is a cosmetic.
[13] The moisturizer of [12], wherein the cosmetic is a leave-on cosmetic.

The moisturizing agent of the present invention can exert a moisturizing ability to retain moisture inside the stratum corneum.
The moisturizing agent of the present invention can also exert a barrier function that suppresses the invasion of foreign substances from the outside into the stratum corneum.
Further, the moisturizing agent of the present invention can be stably blended with a hydrophilic organic solvent and a hydrophobic organic solvent (oil) within an effective concentration range. In particular, the moisturizing agent of the present invention can be stably blended with a hydrophilic organic solvent within an effective concentration range without using an emulsifier.
When the moisturizing agent of the present invention is blended with a hydrophilic organic solvent, the wettability of the formulation is improved, and the compatibility with the skin can be improved.
Furthermore, the moisturizing agent of the present invention is superior in terms of safety to the skin because no cytotoxicity can be observed even when the acyl chain length in the N-acyl amino acid ester is long.
Further, the humectant of the present invention can exhibit a transparent property in a mixed liquid of the humectant of the present invention and a hydrophobic organic solvent (oil) and a mixed liquid of the humectant of the present invention and a hydrophilic organic solvent. Therefore, it is possible to impart a high-class feeling to the appearance, and to avoid visual effects on the skin. Therefore, the moisturizing agent of the present invention has the advantage of being able to reduce, for example, white residue caused by application.

FIG. 1 shows octanoyl valine (Example 3: C8Val), octanoyl alanine (Example 4: C8Ala), glycylproline (Comparative Example 2: Gly-Pro) and valine (Comparative Example 3: Val). FIG. 4 is a diagram showing the amount of amplification of related protein genes (relative value to the amount of amplification of control). The moisturizing-related protein genes tested are, from the left, genes encoding filaggrin, involucrin, transglutaminase 1 (TG1), keratin 10, and loricrin (also in FIG. 2). FIG. 2 shows octanoylleucine (Example 5: C8Leu), octanoylisoleucine (Example 6: C8Ile), octanoylglycine (Example 7: C8Gly), glycylproline (Comparative Example 2: Gly-Pro) and FIG. 3 is a diagram showing the amplification amount (relative value to the control amplification amount) of each moisturizing-related protein gene of valine (Comparative Example 3: Val). Figure 3 shows butanoyl valine (Example 8: C4Val), hexanoyl valine (Example 9: C6Val), octanoyl valine (Example 10: C8Val), decanoyl valine (Example 11: C10Val), lauroyl ( FIG. 3 is a diagram showing the amplification levels (relative values to the control amplification levels) of moisturizing-related protein genes of dodecanoyl)valine (Example 12: C12Val) and methanoylvaline (Example 13: C2Val). The moisturizing-related protein genes tested are, from the left, genes encoding filaggrin, involucrin, and transglutaminase 1 (TG1). Figure 4 shows lauroylvaline (Example 14: C12Val), decanoylvaline (Example 15: C10Val), octanoylvaline (Example 16: C8Val), hexanoylvaline (Example 17: C6Val), butanoylvaline. FIG. 10 is a diagram showing the amount of filaggrin gene amplification of (Example 18: C4Val) and acetylvaline (Example 19: C2Val) (relative value to the amplification amount of the control). Figure 5 shows lauroylalanine (Example 20: C12Ala), decanoylalanine (Example 21: C10Ala), octanoylalanine (Example 22: C8Ala), hexanoylalanine (Example 23: C6Ala), butanoylalanine (Example 24: C4Ala)) shows the amount of amplification of the filaggrin gene (relative value to the amount of amplification of the control). Figure 6 shows lauroylvaline (Comparative Example 4: C12Val), decanoylvaline (Example 25: C10Val), octanoylvaline (Example 26: C8Val), hexanoylvaline (Example 27: C6Val), butanoylvaline. (Example 28: C4Val) and acetylvaline (Example 29: C2Val) cells (percentage of control absorbance). FIG. 7 is a HPLC (manufactured by Shimadzu Corporation) chromatogram (measurement wavelength 210 nm) of N-octanoylalanine production by hydrolysis of C8Ala-iPA in the presence or absence of esterase. On the chromatogram, a peak of N-octanoylalanine is observed at an elution time of 0.6 minutes, and a peak of C8Ala-iPA is observed at an elution time of 1.7 minutes. When C8Ala-iPA was reacted with esterase, the peak at 1.7 minutes decreased and a new peak appeared at 0.6 minutes. FIG. 8 is a HPLC (manufactured by Shimadzu Corporation) chromatogram (measurement wavelength 210 nm) of N-octanoylalanine production by hydrolysis of C8Ala-Et in the presence or absence of esterase. On the chromatogram, an N-octanoylalanine peak is observed at an elution time of 0.6 minutes, and a C8Ala-Et peak is observed at an elution time of 1.2 minutes. When C8Ala-Et is reacted with an esterase, the peak at 1.2 minutes is reduced and a new peak appears at 0.6 minutes.

The present invention provides a moisturizer containing the compound represented by formula (1) above.

A is a straight or branched hydrocarbon group having 1 to 17 carbon atoms. The number of carbon atoms in the hydrocarbon group in A is preferably 16 or less, more preferably 15 or less, still more preferably 13 or less, and even more preferably 11 or less. The number of carbon atoms in the hydrocarbon group in A may also be 2 or more, 3 or more, 4 or more, 5 or more, or 6 or more. More specifically, the number of carbon atoms in the hydrocarbon group in A may preferably be from 2 to 16, more preferably from 3 to 15, even more preferably from 4 to 13, even more preferably from 5 to 11. . The hydrocarbon groups in A are saturated or unsaturated. Such hydrocarbon groups include, for example, alkyls, alkenyls, and alkynyls.

When A is alkyl, alkyl is straight or branched chain alkyl of 1 to 17 carbon atoms. Examples and preferred examples of the number of carbon atoms in alkyl in A are the same as the number of carbon atoms in the above-described hydrocarbon group in A. Examples of alkyl for A include methyl, ethyl, propyl (eg, n-propyl, isopropyl), butyl (eg, n-butyl, iso-butyl, sec-butyl, tert-butyl), pentyl (eg, n- pentyl, iso-pentyl, neo-pentyl, 1-ethylpropyl), hexyl (e.g. n-hexyl, iso-hexyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 2 -ethylbutyl), heptyl (e.g., n-heptyl, 1-methylhexyl, 2-methylhexyl, 3-methylhexyl, 1,1-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2- ethylpentyl), octyl (e.g., n-octyl, 1-methylheptyl, 2-methylheptyl, 3-methylheptyl, 1,1-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 2- ethylhexyl), nonyl (eg, n-nonyl, 3,5,5-trimethylhexyl), decanyl (eg, n-decanyl), undecanyl (eg, n-undecanyl), dodecanyl (eg, n-dodecanyl), tridecanyl ( Examples include n-tridecanyl), tetradecanyl (eg n-tetradecanyl), pentadecanyl (eg n-pentadecanyl), hexadecanyl (eg n-hexadecanyl), and heptadecanyl (eg n-heptadecanyl).

When A is alkenyl, alkenyl is straight or branched chain alkenyl having 2 to 17 carbon atoms. Examples and preferred examples of the number of carbon atoms in alkenyl in A are the same as the number of carbon atoms in the above-described hydrocarbon group in A. Alkenyl in A includes, for example, ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodecenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, and heptadecenyl.

When A is alkynyl, alkynyl is straight or branched alkynyl having 2 to 17 carbon atoms. Examples and preferred examples of the number of carbon atoms in alkynyl in A are the same as the number of carbon atoms in the above-described hydrocarbon group in A. Alkynyl for A includes, for example, ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl, undecynyl, dodecynyl, tridecynyl, tetradecynyl, pentadecynyl, hexadecynyl, and heptadecynyl.

Preferably, A is a straight or branched chain alkyl of 1 to 17 carbon atoms. Examples and preferred examples of the number of carbon atoms in alkyl in A are the same as the number of carbon atoms in the above-described hydrocarbon group in A. Specific examples of such alkyl are given above.

In certain embodiments, the group in A may have 7 to 17 (preferably 7 to 11) carbon atoms. When the number of carbon atoms in the group in A is 7 or more, the compound represented by the above formula (1) can significantly induce the expression of genes encoding various proteins related to improvement of stratum corneum function, It has the advantage that no toxicity can be observed. Such proteins include, for example, proteins capable of producing natural moisturizing factors in skin tissue (e.g., filaggrin, profilaggrin), and proteins capable of participating in skin barrier function (e.g., filaggrin, involucrin, transglutaminase 1, Keratin 10, Loricrin).

In another particular embodiment, the groups in A may have 6 or fewer carbon atoms. When the number of carbon atoms in the group in A is 6 or less, the compound represented by the above formula (1) has a partition coefficient (LogP) in the range of 1 to 4 and a molecular weight of 500 Da or less. It has the advantage of being excellent in cost absorption and being able to be provided in a high concentration in the stratum corneum.

B is a straight or branched hydrocarbon group having 2 to 18 carbon atoms. The number of carbon atoms in the hydrocarbon group in B is preferably 2 to 15, more preferably 2 to 12, even more preferably 2 to 8, even more preferably 2 to 6, particularly preferably may be 2 or 3. The hydrocarbon groups in B are saturated or unsaturated. Such hydrocarbon groups include, for example, alkyls, alkenyls, and alkynyls.

When B is alkyl, alkyl is straight or branched chain alkyl having 2 to 18 carbon atoms. Examples and preferred examples of the number of carbon atoms in alkyl in B are the same as the number of carbon atoms in the hydrocarbon group in B described above. Examples of alkyl for B include ethyl, propyl (eg, n-propyl, isopropyl), butyl (eg, n-butyl, iso-butyl, sec-butyl, tert-butyl), pentyl (eg, n-pentyl, iso-pentyl, neo-pentyl, 1-ethylpropyl), hexyl (e.g. n-hexyl, iso-hexyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 2-ethylbutyl ), heptyl (e.g., n-heptyl, 1-methylhexyl, 2-methylhexyl, 3-methylhexyl, 1,1-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl ), octyl (e.g., n-octyl, 1-methylheptyl, 2-methylheptyl, 3-methylheptyl, 1,1-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 2-ethylhexyl) , nonyl (e.g., n-nonyl, 3,5,5-trimethylhexyl), decanyl (e.g., n-decanyl), undecanyl (e.g., n-undecanyl), dodecanyl (e.g., n-dodecanyl), tridecanyl (e.g., n-tridecanyl), tetradecanyl (e.g. n-tetradecanyl), pentadecanyl (e.g. n-pentadecanyl), hexadecanyl (e.g. n-hexadecanyl), heptadecanyl (e.g. n-heptadecanyl), and octadecanyl (e.g. n-octadecanyl) is mentioned.

When B is alkenyl, alkenyl is straight or branched chain alkenyl having 2 to 18 carbon atoms. Examples and preferred examples of the number of carbon atoms in alkenyl in B are the same as the number of carbon atoms in the hydrocarbon group in B above. Alkenyl in B includes, for example, ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodecenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, and octadecenyl.

When B is alkynyl, alkynyl is straight or branched alkynyl having 2 to 18 carbon atoms. Examples and preferred examples of the number of carbon atoms in alkynyl in B are the same as the number of carbon atoms in the hydrocarbon group in B above. Alkynyl in B includes, for example, ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl, undecynyl, dodecynyl, tridecynyl, tetradecynyl, pentadecynyl, hexadecynyl, heptadecynyl, and octadecynyl.

Preferably, B is a straight or branched chain alkyl of 2 to 18 carbon atoms. Examples and preferred examples of the number of carbon atoms in alkyl in B are the same as the number of carbon atoms in the hydrocarbon group in B described above. Specific examples of such alkyl are given above.

X is a hydrogen atom or a linear or branched hydrocarbon group having 1 to 5 carbon atoms. The number of carbon atoms in the hydrocarbon group in X may be, for example, 1-4. The hydrocarbon group in X is saturated or unsaturated. Such hydrocarbon groups include, for example, alkyls, alkenyls, and alkynyls.

When X is alkyl, alkyl is straight or branched chain alkyl of 1 to 5 carbon atoms. Examples and preferred examples of the number of carbon atoms of alkyl in X are the same as the number of carbon atoms in the above-mentioned hydrocarbon group in X. Examples of alkyl for X include methyl, ethyl, propyl (e.g. n-propyl, isopropyl), butyl (e.g. n-butyl, iso-butyl, sec-butyl, tert-butyl), and pentyl (e.g. n -pentyl, iso-pentyl, neo-pentyl, 1-ethylpropyl).

When X is alkenyl, alkenyl is straight or branched alkenyl having 2 to 5 carbon atoms. Examples and preferred examples of the number of carbon atoms in alkenyl for X are the same as the number of carbon atoms in the above-described hydrocarbon group for X. Alkenyl for X includes, for example, ethenyl, propenyl, butenyl, and pentenyl.

When X is alkynyl, alkynyl is straight or branched alkynyl having 2 to 5 carbon atoms. Examples and preferred examples of the number of carbon atoms of alkynyl in X are the same as the number of carbon atoms in the above-described hydrocarbon group in X. Alkynyl for X includes, for example, ethynyl, propynyl, butynyl, and pentynyl.

Preferably, X is a straight or branched chain alkyl of 1 to 5 carbon atoms. Examples and preferred examples of the number of carbon atoms of alkyl in X are the same as the number of carbon atoms in the above-mentioned hydrocarbon group in X. Specific examples of such alkyl are given above.

In a specific embodiment, the moisturizing agent of the present invention is represented by the above formula (1) [wherein A is a straight or branched hydrocarbon group having 2 to 10 carbon atoms, and B is a It is a straight or branched hydrocarbon group of 2 to 18, and X is a straight or branched hydrocarbon group of 1 to 5 carbon atoms. ] may contain the compound represented by.

Preferably, the compound represented by formula (1) above is a compound represented by any one of formulas (2) to (6) above. Regarding A and B in the compounds represented by the above formulas (2) to (6), the hydrocarbon group (eg, alkyl, alkenyl, alkynyl) and the number of carbon atoms, examples, preferred examples and specific examples are defined by the formula ( 1) is the same as described above.

In certain embodiments, A and B in the compounds represented by formulas (2) to (6) above may each independently have different numbers of carbon atoms and different hydrocarbon groups. For example, A in the compounds represented by (2) to (4) above is a linear or branched hydrocarbon group having 2 to 17 carbon atoms, and represented by (5) and (6) above. A in the compound may be a linear or branched hydrocarbon group having 6 or less carbon atoms, or 7 to 17 (preferably 7 to 11, more preferably 8 to 11) carbon atoms. Further, A in the compounds represented by the above (2) to (4) is a linear or branched hydrocarbon group having 2 to 11 carbon atoms, and represented by the above (5) and (6). A in the compound may be a linear or branched hydrocarbon group having 7 to 11 (preferably 8 to 11) carbon atoms. Further, A in the compounds represented by (2) to (4) above is a straight or branched hydrocarbon group having 5 to 11 carbon atoms, and represented by (5) and (6) above. A in the compound may be a linear or branched hydrocarbon group having 7 to 11 (preferably 8 to 11) carbon atoms.

The compounds represented by formulas (1) to (6) correspond to N-acyl neutral amino acid esters (neutral amino acid derivatives). The compounds represented by formulas (1) to (6) may be optical isomers (L form or D form) with respect to the carbon atom to which all of the nitrogen atom, the carbon atom of the carbonyl group and X are bonded, or a mixture thereof. Preferably, the compounds represented by formulas (1)-(6) are in the L form.

The compound contained in the moisturizing agent of the present invention can be appropriately prepared by reacting an N-acyl neutral amino acid and an alcohol. The reaction of N-acyl neutral amino acids and alcohols can be carried out by general methods for the formation of esters from carboxylic acid compounds and alcohols. Such reactions are well known in the art. For example, the reaction can be carried out by appropriately adding a catalyst. The reaction can also be carried out at a moderate temperature (eg 15-200° C.) for several hours (eg 0.5-8 hours). More specifically, the compound contained in the moisturizing agent of the present invention can be prepared by the method of Production Example A described below. The compounds of the present invention can also be prepared by the method described in JP-A-11-240828, except that the type of N-acyl neutral amino acid and alcohol is changed in the method described in JP-A-11-240828. It may be done in the same way. The compound contained in the moisturizing agent of the present invention can be appropriately purified by any purification method (eg, extraction) used in the field of organic synthesis.

The moisturizer of the present invention is useful, for example, in developing products such as cosmetics, pharmaceuticals and quasi-drugs. The moisturizer of the present invention may be provided in the form of a composition containing the compound represented by formula (1) above and a carrier (eg, a carrier acceptable as a cosmetic, pharmaceutical or quasi-drug). In this case, the moisturizing agent of the present invention, in addition to the components described above, has one or two or more (e.g., two, three, 4) active ingredients may be further included. Examples of such active ingredients include low-molecular-weight compounds.

The term "low molecular weight compound" refers to a compound having a molecular weight of 1500 or less. Small compounds are natural or synthetic compounds. The molecular weight of the low molecular compound may be 1200 or less, 1000 or less, 900 or less, 800 or less, 700 or less, 600 or less, 500 or less, 400 or less, or 300 or less. The molecular weight of the small molecule compound may also be 30 or greater, 40 or greater, or 50 or greater. Examples of low-molecular-weight compounds include amino acids, oligopeptides, vitamins, nucleosides, nucleotides, oligonucleotides, monosaccharides, oligosaccharides, lipids, fatty acids, metabolites thereof, and salts thereof.

Salts include, for example, salts with inorganic acids, salts with organic acids, salts with inorganic bases, salts with organic bases, and salts with amino acids. Salts with inorganic acids include, for example, salts with hydrogen chloride, hydrogen bromide, phosphoric acid, sulfuric acid, and nitric acid. Examples of salts with organic acids include formic acid, acetic acid, trifluoroacetic acid, lactic acid, tartaric acid, fumaric acid, oxalic acid, maleic acid, citric acid, succinic acid, malic acid, benzenesulfonic acid, and p-toluenesulfonic acid. salt of Salts with inorganic bases include, for example, salts with alkali metals (eg, sodium, potassium), alkaline earth metals (eg, calcium, magnesium), and other metals such as zinc, aluminum, and ammonium. . Salts with organic bases include, for example, salts with trimethylamine, triethylamine, propylenediamine, ethylenediamine, pyridine, ethanolamine, monoalkylethanolamine, dialkylethanolamine, diethanolamine, and triethanolamine. Examples of salts with amino acids include salts with basic amino acids (eg, arginine, histidine, lysine, ornithine) and acidic amino acids (eg, aspartic acid, glutamic acid).

When the moisturizing agent of the present invention is a composition, the low-molecular-weight compound that may be further contained in the moisturizing agent of the present invention may be another moisturizing agent. When the moisturizing agent of the present invention is a composition, when the moisturizing agent of the present invention further contains another moisturizing agent, the moisturizing effect can be further improved, and such an embodiment is also preferable.

Other such moisturizing agents include pyrrolidone carboxylic acid, 3-acetyl-2-ethoxycarbonyl-2-methyl-1,3-thiazolidine-4-carboxylic acid, amino acids (e.g. glutamic acid, aspartic acid, arginine, lysine). , histidine, ornithine, glycine, alanine, valine, leucine, isoleucine, serine, threonine, asparagine, glutamine, cysteine, cystine, methionine, phenylalanine, tyrosine, tryptophan, proline, hydroxyproline), glycylglycine, alanylglutamine, dipeptide -2 (valyltryptophan), dipeptide-4 (cystenylglycine), dipeptide-8 (alanylhydroxyproline), dipeptide-9 (glutamyllysine), dipeptide-11 (cystenyllysine), dipeptide-17 (glycylproline) ), dipeptide-19 (leucylglutamic acid), and salts thereof (eg, the salts described above). One or more humectants can be used in combination.

When the moisturizing agent of the present invention is a composition, the moisturizing agent of the present invention may also contain other ingredients such as thickeners, stabilizers, pH adjusters, preservatives, UV inhibitors, fragrances, and pigments. You can The specific types and amounts of these components can be set as appropriate.

Examples of thickeners include carrageenan, dextrin, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyacrylic acid, polymethacrylic acid, carboxyvinyl polymer (carbomer), (acrylic acid/alkyl acrylate ( C10-30)) copolymers, xanthan gum.

Stabilizers include, for example, ascorbic acid, sodium pyrosulfite, EDTA.

Examples of pH adjusting agents include aqueous solutions (buffers) as described above, acidic substances such as hydrochloric acid, and basic substances such as sodium hydroxide.

Preservatives include, for example, ethyl parahydroxybenzoate, sodium benzoate, salicylic acid, sorbic acid, parabens (eg, methylparaben, propylparaben), sodium bisulfite.

Examples of UV inhibitors include UV absorbers (eg, t-butylmethoxydibenzoylmethane, ethylhexyl methoxycinnamate, oxybenzone-3) and UV scattering agents (eg, titanium oxide, zinc oxide).

Fragrances include, for example, limonene, citral, menthol, rose oil and rose oil.

Examples of dyes include organic pigments (e.g., red pigments such as Red No. 201, blue pigments such as Blue No. 404, orange pigments such as Orange No. 203, yellow pigments such as Yellow No. 205, and green pigments such as Green No. 3). , organic lake pigments such as zirconium lake, natural pigments such as chlorophyll), and inorganic pigments (e.g., white pigments such as titanium oxide, colored pigments such as iron oxide, extender pigments such as talc, pearl pigments such as mica). be done.

For example, the moisturizing agent of the present invention is applied to a desired site (eg, skin, hair, body hair, scalp) of a subject (eg, mammals such as humans, animals such as birds, reptiles, etc.) in an effective amount (eg, 0.5%). 001 mg to 100 g) can be used by applying (eg, applying or administering). Preferably, the moisturizer of the present invention is applied to humans. The subject's condition to which the moisturizing agent of the present invention is applied is a healthy condition or an abnormal condition (eg, disease). Such abnormal conditions include, for example, rough skin, dry skin, scales, disordered turnover, and skin diseases (eg, dermatitis such as atopic dermatitis).

The moisturizing agent of the present invention can also contain an effective amount of a compound that induces the expression of genes encoding the above-mentioned various proteins related to improvement of stratum corneum function. Such an effective amount is not particularly limited as long as it is an amount capable of inducing gene expression, but an amount sufficient to induce gene expression is preferable. Such sufficient amount may be, for example, a concentration of 0.01-1000 mM. Such a sufficient amount may also be 500 mM or less, 100 mM or less, 50 mM or less, 10 mM or less, 5 mM or less, or 1 mM or less with a view to reducing compound usage.

The moisturizing agents of the present invention may also be cosmetics or external preparations. The cosmetic or topical preparation of the present invention can be made into any form of formulation that can be applied to a desired site (eg, skin, hair, scalp) according to conventional methods.

The moisturizing agent of the present invention can be preferably used as a cosmetic or external preparation for the skin, hair or scalp. Examples of cosmetics or external preparations for the skin include milky lotions, lotions, creams, gels, beauty essences, and face masks. Cosmetics or external preparations for hair include, for example, emulsions for hair, hair treatments, hair conditioners, shampoos, and hair lotions. Cosmetics or external preparations for the scalp include, for example, hair restorers. Preferred cosmetics include, for example, leave-on cosmetics, emulsions, lotions, creams, gels, beauty essences, and face masks. Preferred external preparations include, for example, ointments, creams, mousses, and gels.

EXAMPLES Next, the present invention will be described in more detail with reference to examples, but the present invention is not limited to the following examples. All of the N-acyl neutral amino acids used as raw materials in the following production examples are L-forms.

Preparation A: Synthesis of N-Acyl Neutral Amino Acid Esters from Alcohols and N-Acyl Neutral Amino Acids .14 mol) was added and mixed. p-Toluenesulfonic acid monohydrate (5.3 g, 0.03 mol) was added to the mixture, a reflux tube was attached so that the alcohol would not evaporate, and the mixture was stirred at a bath temperature of 125° C. for 3 hours. After the product was concentrated by vacuum decompression, the product was diluted with ethyl acetate (600 mL) and washed four times with saturated aqueous sodium bicarbonate (240 mL). The organic phase was further washed twice with ultrapure water (480 mL). Water was removed from the organic layer with sodium sulfate. Furthermore, after removing the sodium phosphate by filtration, the organic layer was concentrated under reduced pressure to obtain the product (acyl neutral amino acid ester).

Production Example 1: Production of N-octanoylalanine isopropyl ester (C8Ala-iPA)

Using N-octanoylalanine (C8Ala, 30.00 g, 0.14 mol) as the N-acyl amino acid and isopropyl alcohol (418 mL, 5.48 mol) as the alcohol, the reaction was performed according to the synthesis method of Production Example A to obtain C8Val. -iPA was obtained (97% yield).

¹ H-NMR (400 MHz, CDCl ₃ , r.t.): δ 6.077-6.093 (1H, d, J=6.4 Hz), 5.015-5.078 (1H, sep, J=6 .4Hz), 4.511-4.583 (1H, quint, J = 7.2Hz), 2.184-2.222 (2H, t, J = 7.6Hz), 1.599-1.669 ( 2H, quint, J = 7.2Hz), 1.372-1.390 (3H, d, J = 7.2Hz) 1.244-1.298 (14H, m), 0.859-0.892 ( 3H, t, J = 6.8 Hz) ppm.

ESI-MS (positive) m/z 258.2 [M+H] ⁺ , 280.1 [M+Na] ⁺

Production Example 2: Production of N-octanoylvaline isopropyl ester (C8Val-iPA)

Using N-octanoyl valine (C8Val, 34.06 g, 0.14 mol) as the N-acyl amino acid and isopropyl alcohol (418 mL, 5.48 mol) as the alcohol, reacted for 8 hours according to the synthesis method of Preparation A. C8Val-iPA was thus obtained (yield 93%).

¹ H-NMR (400 MHz, CDCl ₃ , r.t.): δ 5.955-5.976 (1H, d, J=8.4 Hz), 5.024-5.087 (1H, sep, J=6 .4Hz), 4.528-4.561 (1H, dd), 2.216-2.254 (2H, t, J = 7.2Hz), 2.135-2.182 (1H, sex, J = 6.8 Hz), 1.610-1.765 (2H, quint, J = 7.2 Hz), 1.256-1.314 (14H, m), 0.860-0.949 (9H, m) ppm .

ESI-MS (positive) m/z 286.2 [M+H] ⁺ , 308.1 [M+Na] ⁺

Production Example 3: Production of N-octanoylglycine isopropyl ester (C8Gly-iPA)

Using N-octanoylglycine (C8Gly, 28.18 g, 0.14 mol) as the N-acyl amino acid and isopropyl alcohol (418 mL, 5.48 mol) as the alcohol, the reaction was performed according to the synthesis method of Production Example A to obtain C8Gly. -iPA was obtained (97% yield).

ESI-MS (positive) m/z 244.1 [M+H] ⁺ , 266.1 [M+Na] ⁺

Production Example 4: Production of N-octanoyl isoleucine isopropyl ester (C8Ile-iPA)

Using N-octanoyl isoleucine (C8Ile, 36.03 g, 0.14 mol) as the N-acyl amino acid and isopropyl alcohol (418 mL, 5.48 mol) as the alcohol, the reaction was performed according to the synthesis method of Production Example A to obtain C8Ile. -iPA was obtained (71% yield).

ESI-MS (positive) m/z 300.2 [M+H] ⁺ , 322.2 [M+Na] ⁺

Production Example 5: Production of N-octanoylleucine isopropyl ester (C8Leu-iPA)

Using N-octanoylleucine (C8Leu, 36.03 g, 0.14 mol) as the N-acyl amino acid and isopropyl alcohol (418 mL, 5.48 mol) as the alcohol, the reaction was carried out according to the synthesis method of Production Example A to obtain C8Leu. -iPA was obtained (97% yield).

ESI-MS (positive) m/z 300.2 [M+H] ⁺ , 322.2 [M+Na] ⁺

Production Example 6: Production of N-lauroyl isoleucine isopropyl ester (C12Ile-iPA)

Using N-lauroylisoleucine (C12Ile, 43.89 g, 0.14 mol) as the N-acyl amino acid and isopropyl alcohol (418 mL, 5.48 mol) as the alcohol, the reaction was performed according to the synthesis method of Production Example A to obtain C12Ile-. iPA was obtained (85% yield).

ESI-MS (positive) m/z 356.3 [M+H] ⁺ , 378.2 [M+Na] ⁺

Production Example 7: Production of N-octanoylalanine ethyl ester (C8Ala-Et)

Using N-octanoylalanine (C8Ala, 30.00 g, 0.14 mol) as the N-acylamino acid and ethanol (320 mL, 5.48 mol) as the alcohol, the reaction was performed according to the synthesis method of Production Example A to obtain C8Val-. Et was obtained (90% yield).

¹ H-NMR (400 MHz, CDCl ₃ , r.t.): δ 6.063-6.078 (1H, d, J=6.0 Hz), 4.552-4.624 (1H, quint, J=7 .2Hz), 4.176-4.230 (2H, q, J = 7.2Hz), 2.186-2.224 (2H, t, J = 7.6Hz), 1.599-1.670 ( 2H, quint, J = 7.2Hz), 1.389-1.407 (3H, d, J = 7.2Hz) 1.267-1.302 (11H, m), 0.860-0.892 ( 3H, m) ppm.

ESI-MS (positive) m/z 244.2 [M+H] ⁺ , 266.1 [M+Na] ⁺

Production Example 8: Production of N-octanoylvaline ethyl ester (C8Val-Et)

Using N-octanoylvaline (C8Val, 34.06 g, 0.14 mol) as the N-acyl amino acid and ethanol (320 mL, 5.48 mol) as the alcohol, reacting for 8 hours according to the synthesis method of Production Example A C8Val-Et was obtained (93% yield).

¹ H-NMR (400 MHz, CDCl ₃ , r.t.): δ 5.952-5.973 (1H, d, J=8.4 Hz), 4.557-4.591 (1H, dd), 4. 159-4.241 (2H, m), 2.217-2.255 (2H, t, J=7.2Hz), 2.121-2.202 (1H, sex, J=6.8Hz), 1 .610-1.681 (2H, quint, J=7.2Hz), 1.268-1.303 (11H, m), 0.860-0.952 (9H, m) ppm.

ESI-MS (positive) m/z 272.2 [M+H] ⁺ , 294.1 [M+Na] ⁺

Production Example 9: Production of N-octanoylglycine ethyl ester (C8Gly-Et)

Using N-octanoylglycine (C8Gly, 28.18 g, 0.14 mol) as the N-acyl amino acid and ethanol (320 mL, 5.48 mol) as the alcohol, the reaction was performed according to the synthesis method of Production Example A to give C8Gly- Et was obtained (96% yield).

ESI-MS (positive) m/z 230.2 [M+H] ⁺ , 252.1 [M+Na] ⁺

Production Example 10: Production of N-octanoyl isoleucine ethyl ester (C8Ile-Et)

Using N-octanoyl isoleucine (C8Ile, 36.03 g, 0.14 mol) as the N-acyl amino acid and ethanol (320 mL, 5.48 mol) as the alcohol, the reaction was carried out according to the synthesis method of Production Example A to give C8Ile- Et was obtained (97% yield).

ESI-MS (positive) m/z 286.2 [M+H] ⁺ , 308.2 [M+Na] ⁺

Production Example 11: Production of N-octanoylleucine ethyl ester (C8Leu-Et)

Using N-octanoylleucine (C8Leu, 36.03 g, 0.14 mol) as the N-acyl amino acid and ethanol (320 mL, 5.48 mol) as the alcohol, the reaction was carried out according to the synthesis method of Production Example A to give C8Gly- Et was obtained (98% yield).

ESI-MS (positive) m/z 286.2 [M+H] ⁺ , 308.2 [M+Na] ⁺

Production Example 12: Production of N-lauroyl isoleucine ethyl ester (C12Ile-Et)

Using N-lauroylisoleucine (C12Ile, 43.89 g, 0.14 mol) as the N-acyl amino acid and ethanol (320 mL, 5.48 mol) as the alcohol, C12Ile-Et was prepared by reacting according to the synthesis method of Production Example A. was obtained (yield 91%).

ESI-MS (positive) m/z 342.3 [M+H] ⁺ , 364.3 [M+Na] ⁺

Production Example 13: Production of N-lauroylleucine ethyl ester (C12Leu-Et)

Using N-lauroylleucine (C12Leu, 43.89 g, 0.14 mol) as the N-acyl amino acid and ethanol (320 mL, 5.48 mol) as the alcohol, the reaction was performed according to the synthesis method of Production Example A to give C12Leu-Et. was obtained (88% yield).

ESI-MS (positive) m/z 342.3 [M+H] ⁺ , 364.3 [M+Na] ⁺

Production Example 14: Production of N-Lauroylvaline Ethyl Ester (C12Val-Et)

Using N-lauroylvaline (C12Val, 41.92 g, 0.14 mol) as the N-acyl amino acid and ethanol (320 mL, 5.48 mol) as the alcohol, the reaction was performed for 8 hours according to the synthesis method of Production Example A to obtain C12Val. -Et was obtained (87% yield).

ESI-MS (positive) m/z 328.3 [M+H] ⁺ , 350.2 [M+Na] ⁺

Production Example 15: Production of N-decanoylvaline isopropyl ester (C10Val-iPA)

Using N-decanoyl valine (C10Val, 38 g, 0.14 mol) as the N-acyl amino acid and isopropyl alcohol (418 mL, 5.48 mol) as the alcohol, the reaction was carried out according to the synthesis method of Production Example A to give C10Val-iPA. was obtained (yield 98%).

ESI-MS (positive) m/z 314.5 [M+H] ⁺ , 336.4 [M+Na] ⁺

Production Example 16: Production of N-decanoylalanine isopropyl ester (C10Ala-iPA)

Using N-decanoylalanine (C10Ala, 34.6 g, 0.14 mol) as the N-acyl amino acid and isopropyl alcohol (418 mL, 5.48 mol) as the alcohol, the reaction was carried out according to the synthesis method of Production Example A to obtain C10Ala. -iPA was obtained (97% yield).

ESI-MS (positive) m/z 285.4 [M+H] ⁺ , 307.4 [M+Na] ⁺

Production Example 17: Production of N-decanoylleucine isopropyl ester (C10Leu-iPA)

Using N-decanoyl leucine (C10Leu, 39.9 g, 0.14 mol) as the N-acyl amino acid and isopropyl alcohol (418 mL, 5.48 mol) as the alcohol, C10Leu was reacted according to the synthesis method of Production Example A. -iPA was obtained (97% yield).

ESI-MS (positive) m/z 328.5 [M+H] ⁺ , 350.3 [M+Na] ⁺

Production Example 18: Production of N-hexanoyl valine ethyl ester (C6Val-Et)

Using N-hexanoylvaline (C6Val, 30.14 g, 0.14 mol) as the N-acyl amino acid and ethanol (320 mL, 5.48 mol) as the alcohol, reacting for 8 hours according to the synthesis method of Production Example A C6Val-Et was obtained (82% yield).

ESI-MS (positive) m/z 244.3 [M+H] ⁺ , 266.2 [M+Na] ⁺

Reference Production Examples 1-3: Production of N-lauryl valine isopropyl ester (C12Val-iPA), N-lauryl alanine isopropyl ester (C12Ala-iPA), and N-lauryl alanine ethyl ester (C12Ala-Et) N-lauryl valine isopropyl Ester (C12Val-iPA), N-laurylalanine isopropyl ester (C12Ala-iPA), and N-laurylalanine ethyl ester (C12Ala-Et) were produced according to the description in JP-A-11-240828.

Example 1 Evaluation of Acyl Neutral Amino Acid Esters (1) Evaluation of Compatibility with Hydrophobic Organic Solvents (Oils) 1 g of each sample was weighed out, added to 5 g of liquid paraffin, and stirred at 25° C. for 10 minutes. mixed. After stirring was stopped, the mixture was allowed to stand at 25° C. for 10 minutes, and the state of separation of various samples and liquid paraffin was visually observed and evaluated according to the following criteria.
Very preferred (A): clear solution with no separation observed Preferred (B): clear solution with little separation observed Less preferred (C): locally separated and turbidity not preferred at all: ( D): clear separation occurred

(2) Evaluation of Compatibility with Hydrophilic Organic Solvent 0.5 g of each sample was weighed, added to 10 g of 1,3-butylene glycol, and mixed by stirring at 25° C. for 10 minutes. After stopping the stirring, the mixture was allowed to stand at 25° C. for 10 minutes, and the state of separation of various samples and 1,3-butylene glycol was visually observed and evaluated according to the following criteria.
Very preferred (A): clear solution with no separation observed Preferred (B): clear solution with little separation observed Less preferred (C): locally separated and turbidity not preferred at all: ( D): clear separation occurred

(3) Property evaluation After cooling each compound to 4°C for one day, it was allowed to stand at room temperature (25°C) for 3 days. After 3 days, the properties of each compound were confirmed at room temperature (25°C).
Preferred (A): Liquid at room temperature (25° C.). Therefore, formulations can be easily prepared by simply mixing with other liquid ingredients at room temperature without heating.
Unpreferable (B): It is solid or crystalline at room temperature (25° C.), and it is difficult to prepare a formulation by simply mixing it with other liquid components at room temperature without heating.

As a result, the acyl neutral amino acid ester could be stably blended with a hydrophobic organic solvent (oil) and a hydrophilic organic solvent (Table 1). Therefore, acyl neutral amino acid esters were shown to be superior in terms of production and use.
Acyl neutral amino acid esters also facilitated formulation (Table 1). Therefore, acyl neutral amino acid esters were shown to be excellent in handleability.

Example 2 Evaluation of Esterase Hydrolysis of Acyl Neutral Amino Acid Ester 5 mg of acyl neutral amino acid ester was added to 1×PBS buffer, and 2.15 μl (0.2 wt %) of Triton-X100 (manufactured by Aldrich) was added. It was mixed uniformly by post-ultrasound irradiation. A buffer solution of esterase (Esterase from porcine liver, manufactured by Sigma-Aldrich) was added to the mixture so that 5 Units of esterase per molecule of acylamino acid was added, and the mixture was stirred and mixed. This mixture was shaken and stirred for 24 hours in a shaker heated to 30° C. to react. The reactants were centrifuged at 12000×g and the supernatant was analyzed by high performance liquid chromatography (Shimadzu). As a control, only buffer was added instead of the esterase solution. The esterase used (Esterase from porcine liver) was used as a model for the esterase enzyme present in human skin [Bonina, F. et al. P. et al. , Eur. J. Pharm. Sci. 14, 123-134 (2001); Wong, O.; et al. , Int. J. Pharm. , 52, 191-202 (1989); et al. , J. Control Release. 104, 41-49 (2005)].

When the ester bond of the acyl neutral amino acid ester is hydrolyzed by esterase, the peak area of the acyl neutral amino acid ester decreases on the HPLC chromatogram, and a new peak is confirmed at the same elution time as that of the acyl neutral amino acid. . From the appearance of this peak, it can be concluded that acyl neutral amino acid ester was hydrolyzed by esterase to produce acyl neutral amino acid.

As test substances, the compounds produced in Production Examples 1 to 18 and Reference Production Examples 1 to 3 were used.

In the presence of an esterase, C6Val-Et, C8Ala-iPA, C8Val-iPA, C8Gly-iPA, C8Ile-iPA, C8Leu-iPA, C8Ala-Et, C8Val-Et, C8Gly-Et, C8Ile-Et, C8Leu-Et, C12Ile-Et, C10Ala-iPA, C10Val-iPA, C10Leu-iPA, C12Ala-Et, C12Ala-iPA, C12Val-Et, C12Val-iPA, C12Ile-iPA, C12Leu-Et are all hydrolyzed to form C6Val, C8Ala, C8Val, C8Gly, C8Ile, C8Leu, C10Ala, C10Val, C10Leu, C12Ala, C12Val, C12Ile, C12Leu were produced.

Examples 3 to 7 and Comparative Examples 2 to 3: Comparison of amplification amounts of genes such as moisturizing-related proteins by octanoyl neutral amino acids ), adjusted to a concentration of 30×10 ⁴ cells/mL, added 2 mL of the adjusted culture medium (ie, 60×10 ⁴ cells/well) to a 6-well plate, and cultured at 37° C. for 1 day. Samples (octanoyl valine (Example 3: C8Val), octanoyl alanine (Example 4: C8Ala), octanoyl leucine (Example 5: C8 Leu), octanoyl isoleucine (Example 6: C8 Ile), octanoyl glycine ( Example 7: C8Gly), glycylproline (Comparative Example 2: Gly-Pro), and valine (Comparative Example 3: Val)) were added at concentrations shown in each figure, and reacted for 48 hours. After the reaction, total RNA was extracted from the cells, reverse transcription was performed, and the amplification amount of target genes (filagrin, involucrin, transglutaminase (TG) 1, keratin 10, and loricrin) was measured by real-time PCR (n = 2), compared with the amplified amount of the control (no addition). GAPDH was used as a correction gene. The comparative CT method was used for calculation, TaqMan (registered trademark) Gene Expression was used for primers and probes, and FAM was used for fluorescent dye. The results are shown in Figures 1-2.

As is clear from FIGS. 1 and 2, the amount of amplification of each target gene in Examples 3-7 was higher than in Comparative Examples 2 and 3. These results indicate that acyl neutral amino acids can promote the production of moisturizing-related proteins.

Examples 8 to 13: Comparison of amount of amplification of genes such as moisturizing-related proteins by acylvaline 10: C8Val), decanoylvaline (Example 11: C10Val), lauroylvaline (Example 12: C12Val) and acetylvaline (Example 13: C2Val), and the target genes were filaggrin, involucrin, and The procedure was carried out in the same manner as in Example 3, except that transglutaminase (TG) was 1. The results are shown in FIG.

As is clear from FIG. 3, in all of Examples 8 to 13, the amount of amplification of each target gene was higher than that of the control. These results indicate that an acyl-neutral amino acid having an acyl group of 2 to 12 carbon atoms or a salt thereof can promote the production of moisturizing-related proteins regardless of the type of acyl group.

Examples 14 to 24: Comparison of Amount of Filaggrin Gene Amplification by Acylvaline or Acylalanine Normal human epidermal keratinocytes NHEK (NB) (manufactured by Kurabo Industries) were precultured in HuMedia-KG2 (manufactured by Kurabo Industries), and 15×10 ⁴ cells/cell were obtained. 2 mL of the adjusted culture medium (ie, 30×10 ⁴ cells/well) was added to a 6-well plate and cultured at 37° C. for 1 day. Samples (Lauroylvaline (Example 14: C12Val), Decanoylvaline (Example 15: C10Val), Octanoylvaline (Example 16: C8Val), Hexanoylvaline (Example 17: C6Val), Butanoylvaline (Example 17: C6Val) Example 18: C4Val), acetylvaline (Example 19: C2Val), lauroylalanine (Example 20: C12Ala), decanoylalanine (Example 21: C10Ala), octanoylalanine (Example 22: C8Ala), hexanoyl Alanine (Example 23: C6Ala) and butanoylalanine (Example 24: C4Ala)) were added at concentrations shown in each figure, and reacted for 24 hours. After the reaction, total RNA was extracted from the cells, reverse transcription was performed, and the amplification amount of the target gene (filagrin) was measured by real-time PCR (n = 2). compared with this value. GAPDH was used as a correction gene. The comparative CT method was used for calculation, TaqMan (registered trademark) Gene Expression was used for primers and probes, and FAM was used for fluorescent dye.

The following evaluations were made according to the amount of amplification. The results are shown in Tables 2-3 and Figures 4-5.
Very preferred (A): 200% or more Preferred (B): 100% or more and less than 200% Less preferred (C): 50% or more and less than 100% Not preferred at all (D): Less than 50%

Considering the above results of Examples 2 to 24 (Tables 2 and 3, Figures 1 to 8), acyl neutral amino acid esters were able to improve the expression-promoting action of moisturizing-related proteins such as filaggrin. , it is thought that the moisturizing ability to retain moisture inside the stratum corneum can be improved. In addition, since the acyl neutral amino acid ester was able to improve the expression-promoting action of proteins related to skin barrier ability, it is thought that the barrier ability to suppress invasion of foreign matter can be improved. In addition to these results, taking into account the results of Example 1 (Table 1), acyl neutral amino acid esters are excellent in handling properties and exhibit sufficient keratin function-improving ability due to their action inside the stratum corneum. It is possible.

Examples 25 to 31: Comparison of Cytotoxicity of Acylvaline and Acylvaline Ester Human normal epidermal keratinocytes NHEK (NB) (manufactured by Kurabo Industries) were precultured in HuMedia-KG2 (manufactured by Kurabo Industries) at 15×10 ⁴ cells/mL. 2 mL of the adjusted culture medium (ie, 30×10 ⁴ cells/well) was added to a 6-well plate and cultured at 37° C. for 1 day. Samples (Lauroylvaline (Comparative Example 4: C12Val), Decanoylvaline (Example 25: C10Val), Octanoylvaline (Example 26: C8Val), Hexanoylvaline (Example 27: C6Val), Butanoylvaline (Example 27: C6Val) Example 28: C4Val), acetylvaline (Example 29: C2Val), lauroylvaline-iPA (Example 30: C12Val-iPA), and lauroylvaline-Et (Example 31: C12Val-Et) at concentrations shown in each figure. After that, the medium was discarded and replaced with 2 mL/well of Neutral Red solution, and cultured for another 2 hours.The Neutral Red solution was Neutral Red (Kurabo) in HuMedia-KG2 medium/ The cells were diluted to 1/100 with a mixed solution of physiological saline (2:1), and the cells stained with the Neutral Red solution were washed once with PBS (-), and then washed with 1% CaCl ₂ /1% formalin. After fixation with 1.5 mL/well of solution for 1 minute, 1.5 mL/well of 1% acetic acid/50% ethanol solution was added for extraction at room temperature for 20 minutes, and absorbance at 540/630 nm was measured with a plate reader.

The following evaluations were made according to the cell viability. Results are shown in Table 4 and FIG.
Very preferred (A): 95% or more Preferred (B): 75% or more and less than 95% Less preferred (C): 50% or more and less than 75% Not preferred at all (D): Less than 50%

The results of Examples 14-24 demonstrate that each acyl neutral amino acid can amplify the filaggrin gene. Further, the results of Examples 25 to 31 and Comparative Example 4 show that the acyl neutral amino acids and acyl neutral amino acid esters of Examples 25 to 31 have low cytotoxicity, whereas the acyl neutral amino acid of Comparative Example 4 has low cytotoxicity. It shows high cytotoxicity. Interestingly, N-acylamino acid esters with long acyl chains were not cytotoxic, unlike N-acyl-amino acids with similarly long acyl chains. Summarizing the above cytotoxicity evaluations, it can be seen that acyl neutral amino acid esters are preferred over acyl neutral amino acids.

Example 32: Compatibility Test of Acyl Neutral Amino Acid Esters and Hydrophilic Organic Solvents The components listed in Table 5 were weighed into a 1.5 mL Eppendorf tube, stirred with a vortex mixer (Scientific Industries Vortex Genie 2) and mixed with hands. The sample was vigorously shaken up and down using a sample to uniformly agitate, and the state after agitation was visually observed. Criteria for judgment were A for uniform transparent dissolution, and B for dispersion of oil droplets and confirmation of scattered light. The evaluation was carried out under conditions of storage at 25°C, -5°C, and 45°C for one day.

As a result, acyl neutral amino acid esters had high solubility stability in hydrophilic organic solvents within the effective concentration range (Tables 5-1 and 5-2). Therefore, it was shown that the acyl neutral amino acid ester can be stably mixed with a hydrophilic organic solvent within an effective concentration range without using an emulsifier.

Example 33: Compatibility Improvement Test of Acyl Neutral Amino Acid Ester and Hydrophilic Organic Solvent The samples used in the above compatibility test were used for evaluation (Table 6). Using a polydropper, one drop of the sample was dropped onto a slide glass, and the spread of the sample after standing for 5 minutes was measured using a ruler. It is considered that the wider the spread of the sample is, the easier it is to blend in during application, which is preferable for practical use. The degree of spreading was evaluated by the following equation, where a is the measured value of the polyhydric alcohol to which the acyl neutral amino acid ester or oil was added, and b is the measured value of the polyhydric alcohol alone. The degree of spread was evaluated on a 4-grade scale.

Degree of spread: (a/b) x 100
A: 150% or more B: 130% or more and less than 150% C: 100% or more and less than 130% D: less than 100%

As a result, acyl neutral amino acid esters were able to improve compatibility with hydrophilic organic solvents (Table 6). Therefore, it was shown that the acyl neutral amino acid ester can improve the wettability of the formulation by blending with the hydrophilic organic solvent, and improve the compatibility with the skin.

Example 34: Improvement of profilaggrin production rate by octanoylvaline (C8Val) Cultured under CO2 and saturated steam. Confluent cells were seeded at 15×10 ⁴ (cells/well) in 6-well plates. After one day of culture at 37°C, the medium was changed to HuMedia-KG2 medium supplemented with 100 µM octanoyl valine (C8Val) and 1.3 mM CaCl ₂ , and the medium was changed in the same way every 1 to 2 days, and cultured for 6 days. did. After culturing, the medium was removed, washed twice with cold PBS (-), and an extract (M-PER (Thermo Fisher Scientific)) added with a protease inhibitor and a phosphatase inhibitor (EzRIPA Lysis (ATTO)). was added to each well at 200 μL/well, and shaken at 200 rpm for 5 minutes with a shaker (manufactured by TAITEC). Scrape the cells with a scraper, add 200 μL of denaturant (EzApply (manufactured by ATTO)) to the total amount of liquid including the precipitate, and heat at 1100 rpm and 100 ° C. for 5 minutes with a shaker (manufactured by Bioer Technology). shaken. The resulting solution was centrifuged at 10000 G in a centrifuge (manufactured by Tomy Seiko Co., Ltd.) at 4°C for 10 minutes, and the supernatant was used as a sample for Western blotting.

Next, 20 μL of each western blotting sample (profilaggrin) per gel lane, or 5 μL of a 50-fold diluted solution of the western blotting sample prepared in (1) (GAPDH) was applied to e-PAGEL 7.5% (ATTO). (manufactured by ATTO) and electrophoresed at a constant current of 21 mA for 1 hour in an electrophoresis apparatus (manufactured by ATTO). After electrophoresis, the gel was transferred to a PVDF membrane, washed with TBS-T (manufactured by ATTO), and blocked using EzBlock BSA (manufactured by ATTO). Next, an antigen-antibody reaction was performed with a primary antibody solution (Anti Filaggrin (AKH1) antibody (manufactured by Santa Cruz Biotechnology) (Profilaggrin), Anti GAPDH (6C5) antibody (manufactured by Santa Cruz Biotechnology) (GAPDH)). rice field. Labeled with a secondary antibody (Anti-IgG (H + L), Mouse, Goat-poly, HRP (manufactured by KPL)), band intensity (250-400 kDa (profilaggrin ), 37 kDa (GAPDH)) were quantified (n=2). The expression level of profilaggrin was normalized by the expression level of GAPDH, and the production level of the control (octinoylvaline (C8Val)-free) was set as 100% for comparison.

The obtained results were evaluated as follows. Table 7 shows the results.
Very preferred (A): 250% or more Preferred (B): 150% or more and less than 250% Less preferred (C): 100% or more and less than 150% Not preferred at all (D): Less than 100%

From the above, it was clarified that octanoylvaline (C8Val) improves profilaggrin protein production.

Example 35: Evaluation of partition coefficient of acyl neutral amino acid ester Calculated using calculation software.

From Table 8, the LogP value of acyl neutral amino acid esters such as C6Ala-iPA, C6Val-Et, and C6Ile-Et is within the range of 1 to 4, and it has a property of superior percutaneous absorbability. can be understood (Schneider, M. et al., Dermatoendocrinol., 1, 197-206 (2009)).

The moisturizer of the present invention is useful, for example, for the development of products such as cosmetics, pharmaceuticals and quasi-drugs, and as products such as cosmetics, pharmaceuticals and quasi-drugs.

Claims (25)

Formula (1) below:

[In the formula,
A is a linear or branched hydrocarbon group having 1 to 17 carbon atoms,
B is a linear or branched hydrocarbon group having 2 to 18 carbon atoms,
X is a hydrogen atom or a linear or branched hydrocarbon group having 1 to 5 carbon atoms. ] A moisturizing agent containing a compound represented by The compound represented by the formula (1) is represented by the following formulas (2) to (6):

[In the formula,
A and B are each the same as in formula (1) above. ] The moisturizing agent according to claim 1, which is a compound represented by any one of 3. The moisturizing agent according to claim 2, wherein the compound represented by formula (1) is the compound represented by formula (2). 3. The moisturizing agent according to claim 2, wherein the compound represented by formula (1) is the compound represented by formula (3). 3. The moisturizing agent according to claim 2, wherein the compound represented by formula (1) is the compound represented by formula (4). 3. The moisturizer according to claim 2, wherein the compound represented by formula (1) is the compound represented by formula (5). 3. The moisturizing agent according to claim 2, wherein the compound represented by formula (1) is the compound represented by formula (6). The moisturizing agent according to any one of claims 1 to 7 , wherein A is a straight or branched hydrocarbon group having 1 to 11 carbon atoms. The moisturizing agent according to any one of claims 1 to 7 , wherein A is a straight or branched hydrocarbon group having 7 to 17 carbon atoms. The moisturizing agent according to claim 8 or 9, wherein A is straight or branched alkyl having 7 to 11 carbon atoms. 11. A moisturizer according to claim 10, wherein A is straight chain alkyl of 7 carbon atoms. The moisturizing agent according to any one of claims 1 to 11 , wherein B is a straight or branched hydrocarbon group having 2 to 8 carbon atoms. 13. The moisturizing agent according to claim 12 , wherein B is a straight or branched hydrocarbon group having 2 or 3 carbon atoms. A moisturizer according to any one of claims 1 to 13 , wherein the hydrocarbon groups in A, B and X are alkyl. 4. The compound according to claim 3, wherein the compound represented by formula (2) is N-octanoyl isoleucine ethyl ester, N-octanoyl isoleucine isopropyl ester, N-lauroyl isoleucine ethyl ester, or N-lauroyl isoleucine isopropyl ester. Moisturizer. 5. The compound according to claim 4, wherein the compound represented by formula (3) is N-octanoylleucine ethyl ester, N-octanoylleucine isopropyl ester, N-lauroylleucine ethyl ester, or N-decanoylleucine isopropyl ester. moisturizer. The moisturizing agent according to claim 5, wherein the compound represented by formula (4) is N-octanoylalanine ethyl ester, N-octanoylalanine isopropyl ester, or N-decanoylalanine isopropyl ester. The compound represented by the formula (5) is N-hexanoyl valine ethyl ester, N-octanoyl valine ethyl ester, N-octanoyl valine isopropyl ester, N-decanoyl valine isopropyl ester, or N-lauroyl valine ethyl 7. A moisturizer according to claim 6, which is an ester. The moisturizer according to claim 7, wherein the compound represented by formula (6) is N-octanoylglycine ethyl ester or N-octanoylglycine isopropyl ester. The moisturizing agent according to any one of claims 1 to 19 , which has an effect of improving stratum corneum function. 21. The moisturizing agent according to any one of claims 1 to 20 , which has an action of promoting the production of one or more proteins selected from the group consisting of profilaggrin, filaggrin, involucrin, transglutaminase 1, keratin 10, and loricrin. The moisturizing agent according to any one of claims 1 to 21 , which is a composition containing said compound at a concentration of 0.01 to 1000 mM. The moisturizing agent according to any one of claims 1 to 22 , which is an external skin preparation. The moisturizing agent according to any one of claims 1 to 22 , wherein the moisturizing agent is a cosmetic. 25. The moisturizing agent according to claim 24 , wherein the cosmetic is a leave-on cosmetic.

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