JP2002316916A - Hair growing agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a hair growing agent containing a phosphatidic acid as an active ingredient. SOLUTION: This hair growing agent contains the phosphatidic acid expressed by formula (I) as the active ingredient. (R<1> is an alkyl group, an alkenyl group, an alkanoyl group or an alkenoyl group, in the case where R<1> is an alkyl group or an alkenyl group then R<2> means an alkyl group, an alkenyl group, an alkanoyl group or an alkenoyl group, in the case where R<1> is an alkanoyl group or an alkenoyl group then R<2> is an alkyl group or an alkenyl group).

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a hair restorer containing phosphatidic acid as an active ingredient.

[0002]

BACKGROUND OF THE INVENTION The invention relating to hair-related cosmetics or pharmaceuticals containing phosphatidic acid includes phosphatidic acid having an odd-numbered straight-chain carbon chain fatty acid residue, namely,
Formula (II)

[0003]

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Wherein R ³ and R ⁴ represent an aliphatic hydrocarbon group, and at least one of R ³ and R ⁴ is a straight-chain aliphatic hydrocarbon group having an even carbon chain length. A hair restorer containing phosphatidic acid is known (JP-B-63-41363). JP-A-61-7205 describes a cell activator containing phosphatidic acid having two branched fatty acid residues as an active ingredient. However, a hair restorer containing a phosphatidic acid having at least one ether group is not known. Also, a fatty acid having an odd number of carbon chains and biotin or vitamin B
Cell activators containing ₁₂ as an active ingredient are known (JP-A-61-15809).

[0005] It is known that phosphatidic acid is used as a liposome preparation base to be added to minoxidil which is an active ingredient for hair growth (US 5030).
442). A mixture of several phospholipids containing phosphatidic acid has an effect of suppressing hair loss (WO 97/0
9989), and also a hair restorer comprising a mixture of phospholipids is known (DE 322216, DE 411334).
6).

[0006] Hair restorers containing proanthocyanidins are described in WO 96/00561. Also, tocopherol [Hair Medicine, p. 283 (1987), Bunkodo;
Hair Science, p. 80 (1986), Japan Hair Science Church], pantothenic acid and biotin [Fragrance Journal, pp. 95-103 (1989), Fragrance Journal], and a protein kinase C-specific inhibitor [Skin Skin Pharmacology and Applied Skin Physiology
Skin Physiology), 13 , 133-142 (2000)] are known to have a hair-growth effect.

Japanese Patent Application Laid-Open No. Hei 11-263711 discloses that
O-hexadecyl-2-O-methylglycerol is described in terms of hair growth activity. However, phosphatidic acid represented by the formula (I) described below, proanthocyanidin, tocopherol, tocopherol derivative, pantothenic acid, pantothenic acid derivative, protein kinase C
Hair restorer containing as an active ingredient at least one component selected from the group consisting of a specific inhibitor or a pharmaceutically acceptable salt thereof, and biotin; phosphatidic acid, proanthocyanidin, tocopherol, tocopherol derivative, pantothene A hair restorer comprising, as an active ingredient, at least one component selected from the group consisting of an acid, a pantothenic acid derivative and a protein kinase C-specific inhibitor or a pharmaceutically acceptable salt thereof; and phosphatidic acid and biotin. , Proanthocyanidins, tocopherols,
A hair restorer comprising as an active ingredient at least one ingredient selected from the group consisting of a tocopherol derivative, a pantothenic acid, a pantothenic acid derivative and a protein kinase C-specific inhibitor or a pharmacologically acceptable salt thereof is known. Not.

[0008]

SUMMARY OF THE INVENTION It is an object of the present invention to provide a hair restorer containing phosphatidic acid as an active ingredient and having an excellent hair growth / growth effect. The above-mentioned phosphatidic acid, proanthocyanidin and tocopherol are also provided. Containing, as an active ingredient, one or more components selected from the group consisting of tocopherol derivatives, pantothenic acid, pantothenic acid derivatives, protein kinase C-specific inhibitors or pharmacologically acceptable salts thereof, and biotin. And a hair restorer characterized by the following.

[0009]

Means for Solving the Problems The present invention relates to the following. (1) Formula (I)

[0010]

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(Wherein, R ¹ represents alkyl, alkenyl, alkanoyl or alkenoyl, and when R ¹ represents alkyl or alkenyl, R ² represents alkyl, alkenyl,
A phosphatidic acid represented by alkanoyl or alkenoyl, and when R ¹ represents alkanoyl or alkenoyl, R ² represents alkyl or alkenyl) as an active ingredient.

(2) The hair restorer according to the above (1), wherein R ¹ is alkyl or alkenyl and R ² is methyl.

(3) The hair restorer according to the above (1), wherein R ¹ is alkanoyl or alkenoyl, and R ² is methyl.

(4) The hair restorer according to the above (1), wherein R ¹ is alkyl or alkenyl and R ² is acetyl.

(5) The hair restorer according to the above (1), wherein R ¹ is octadecenoyl or hexadecyl, and R ² is methyl.

(6) The hair restorer according to the above (1), wherein R ¹ is hexadecyl and R ² is acetyl.

(7) The phosphatidic acid according to any of (1) to (6) above, proanthocyanidin, tocopherol, a tocopherol derivative, pantothenic acid,
A hair restorer comprising, as an active ingredient, one or more ingredients selected from the group consisting of a pantothenic acid derivative, a protein kinase C-specific inhibitor or a pharmaceutically acceptable salt thereof, and biotin.

(8) The hair restorer according to any one of the above (1) to (6), comprising proanthocyanidin.

(9) Proanthocyanidins are procyanidin B-1, procyanidin B-2, procyanidin B-
3. The hair restorer according to the above (8), which is one or more proanthocyanidins selected from the group consisting of procyanidin C-1 and procyanidin C-2.

(10) The above (1) to (1), which contain tocopherol or a tocopherol derivative.
(6) The hair restorer according to any one of (8) and (9).

(11) the tocopherol or tocopherol derivative is selected from the group consisting of dl-α-tocopherol, d-α-tocopherol, dl-α-tocopherol acetate, d-α-tocopherol acetate and dl-α-tocopherol nicotinate The hair restorer according to the above (10), which is one or more tocopherols or tocopherol derivatives.

(12) The above (1) to (1), which contain pantothenic acid or a pantothenic acid derivative.
(6) The hair restorer according to any one of (8) to (11). (13) One or more pantothenic acids or pantothenic acids or pantothenic acid derivatives, wherein the pantothenic acid or pantothenic acid derivative is selected from the group consisting of calcium pantothenate, sodium pantothenate, D-pantothenyl alcohol, DL-pantothenyl alcohol and pantothenyl ethyl ether The hair restorer according to the above (12), which is an acid derivative.

(14) The above (1) to (6) or (8) to (1), which comprises a protein kinase C-specific inhibitor or a pharmacologically acceptable salt thereof.
The hair restorer according to any one of 3).

(15) Protein kinase C-specific inhibitors are calhostin C, hexadecylphosphocholine, palmitoyl-DL-carnitine and polymyxin B
One or more protein kinases selected from
The hair restorer according to the above (14), which is a C inhibitor.

(16) The hair restorer according to any one of the above (1) to (6) or (8) to (15), which contains biotin.

(17) The hair restorer according to any one of the above (1) to (16), which does not substantially contain minoxidil.

(18) one selected from the group consisting of phosphatidic acid, proanthocyanidin, tocopherol, a tocopherol derivative, pantothenic acid, a pantothenic acid derivative, and a protein kinase C-specific inhibitor or a pharmaceutically acceptable salt thereof A hair restorer containing the above components as active ingredients.

(19) selected from the group consisting of phosphatidic acid and biotin, and proanthocyanidins, tocopherols, tocopherol derivatives, pantothenic acid, pantothenic acid derivatives, and protein kinase C-specific inhibitors or pharmacologically acceptable salts thereof. A hair restorer containing one or more components as an active ingredient.

[0029]

BEST MODE FOR CARRYING OUT THE INVENTION In the definition of each group of the formula (I),
As the alkyl, for example, 1 to 24 carbon atoms, preferably 7 to
19 alkyls, more specifically, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, Heneicosyl, docosyl, tricosyl, tetracosyl and the like. As alkenyl, for example, having 2 to 2 carbon atoms
4, preferably 7 to 19 alkenyl, more specifically vinyl, allyl, butenyl, 2-pentenyl, 4-pentenyl, hexenyl, heptenyl, octenyl, nonenyl,
Decenyl, undecenyl, dodecenyl, tridecenyl,
Examples include tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, nonadecenyl, eicosenyl, heneicosenyl, docosenyl, tricosenyl, tetracosenyl, and the like. The number and position of unsaturated bonds in alkenyl are not particularly limited. Examples of the alkanoyl include alkanoyl having 2 to 24 carbon atoms, preferably 7 to 20 carbon atoms, more specifically, acetyl, propionyl, butyryl, valeryl, hexanoyl, heptanoyl, octanoyl, nonanoyl, decanoyl, undecanoyl, dodecanoyl, tridecanoyl, and tetradecanoyl. Decanoyl, pentadecanoyl, hexadecanoyl, heptadecanoyl, octadecanoyl, nonadecanoyl, eicosanoyl, heneicosanoyl, docosanoyl, tricosanoyl, tetracosanoyl and the like. Alkenoyl, for example, having 3 to 24 carbon atoms,
Preferably 7 to 20 alkenoyls, more particularly propenoyl, butenoyl, 2-pentenoyl, 4-pentenoyl, hexenoyl, heptenoyl, octenoyl, nonenoyl, decenoyl, undecenoyl, dodecenoyl, tridecenoyl, tetradecenoyl, pentadecenoyl, pentadecenoyl, pentadecenoyl, pentadecenoyl, pentadecenoyl, pentadecenoyl Cis-9-
Octadecenoyl, cis-9-cis-12-octadecadienoyl, 9,12,15-octadecatrienoyl, nonadecenoyl, 5,8,11,14-eicosatetraenoyl, heneicosenoyl, docosenoyl, tricosenoyl, And tetracosenoyl. The number and position of the unsaturated bonds contained in the alkenoyl are not particularly limited.

The phosphatidic acid used in the present invention is
It can be obtained mainly by chemical synthesis. That is, it can be obtained by introducing a fatty acid or an alcohol into position 1 of glycerin, then introducing a fatty acid or an alcohol into position 2 of glycerin, and phosphorylating position 3 of glycerin. In each synthesis step, a protecting group is introduced as necessary. For example, the desired phosphatidic acid is added dropwise to a mixture of 1 to 5 equivalents of phosphorus oxychloride, 1 to 5 equivalents of triethylamine and hexane, and 1 to 5 equivalents of a glycerol derivative in an inert solvent is added to the solvent. Preferably, the mixture is stirred under ice cooling for 10 to 20 hours, further added with purified water, stirred at room temperature for 1 hour, washed with an organic layer, and concentrated under reduced pressure to obtain the organic layer. Examples of the inert solvent include chloroform and hexane. Further, by enzymatic degradation of a phospholipid derivative containing a fatty acid residue or an alcohol residue, for example, when a phosphatidylcholine derivative is used as a raw material, a phosphate bond with choline is hydrolyzed using an enzyme such as phospholipase D. Can also be used to obtain the desired phosphatidic acid [Okuyama Harumi, Lipid Biochemistry (Biochemistry Experiment Course 3), The Japanese Biochemical Society, 289
Page (1974), Tokyo Doujinshi]. However, the production method is not limited to these, and the above phosphatidic acid obtained by any method can be used in the present invention.

The proanthocyanidins used in the present invention have the formula (III)

[0032]

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Wherein R ⁵ and R ⁶ are the same or different and each represent a hydrogen atom or a hydroxyl group.
-Refers to a group of compounds polymerized using an all derivative as a structural unit. Specific examples of the flavan-7-ol derivative include catechin, epicatechin, gallocatechin, epigallocatechin, afzeletin, epiafuzeletin and the like, and all of these optical isomers are included, but in the present invention, epicatechin or catechin Is more preferably used.

The binding mode of the flavan-7-ol derivative represented by the formula (III) includes any binding mode.
For example, a dimer in which two flavan-7-ol derivatives are polymerized is represented by the formula (IV)

[0035]

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Wherein R ⁷ and R ⁸ and R ^7a and R ^8a
Have the same meanings as those of R ⁵ and R ⁶ ), and examples of the trimer or higher polymer include those obtained by combining the same or different bonding modes. . The proanthocyanidin used in the present invention may be a dimer or more of a flavan-7-ol derivative, preferably 2 to 10 mer, more preferably 2 to 5 mer, and still more preferably 2 to 3 mer. Body. As the dimer of the flavan-7-ol derivative, for example, a combination of epicatechin and catechin such as epicatechin- (4β → 8) -catechin, and epicatechin such as epicatechin- (4β → 8) -epicatechin Dimers, catechin dimers such as catechin- (4α → 8) -catechin and the like, and trimers of flavan-7-ol derivatives include, for example, epicatechin- (4β → 8) -epicatechin Epicatechin 3 such as-(4β → 8) -epicatechin
Catechins such as catechin- (4α → 8) -catechin- (4α → 8) -catechin, epicatechin- (4β → 8) -epicatechin- (4β → 8) -catechin A mixed trimer of catechin and catechin is exemplified.

Compounds obtained by adding sugars such as gallic acid, glucose and rhamnose to these proanthocyanidins are also included in the proanthocyanidins used in the present invention. Proanthocyanidins include grape, apple, barley, oyster, coconut, cacao, pine, kidney bean, peanut, etc.
It can be obtained by extracting and purifying from various plants such as barley, oyster, palm, cacao, pine, adzuki, peanut and the like, and can also be obtained by chemical synthesis.

For example, the extraction and purification of proanthocyanidins from plants can be performed by the following known methods. The raw material of a plant, such as fruit, seed, leaf, stem, root, rhizome, etc., is collected at an appropriate time, and then subjected to a drying step as it is or usually by air drying to obtain an extraction raw material. When proanthocyanidins are extracted from dried plants, a known method [Chemical & Pharmaceutical Bulletin, 3 , 3218]
(1990) and 40 , 889-898 (1992)].

That is, after the raw material is pulverized or shredded, extraction is performed using a solvent. As the extraction solvent, water,
Hydrophilic or lipophilic solvents such as alcohols such as ethanol, methanol and isopropyl alcohol, ketones such as acetone and methyl ethyl ketone, and esters such as methyl acetate and ethyl acetate can be used alone or as a mixed solvent. Extraction temperature is usually 0 ~ 100 ℃,
Preferably it is 5-50 degreeC. The extraction time is about 1 hour to 10 days, and the amount of the solvent is usually 1 to 30 times, preferably 5 to 10 times the weight of the dry raw material. The extraction operation is performed by stirring or immersion, and is repeated two or three times as necessary.

An extract obtained by removing an insoluble residue from the crude extract obtained by the above operation by filtration or centrifugation,
Alternatively, the method for purifying proanthocyanidins from plant sap or sap may be any known separation and purification method, such as two-phase solvent partitioning, column chromatography, or preparative high-performance liquid chromatography. Are preferably used alone or in combination. For example, as a two-phase solvent partitioning method, a method of extracting and removing oil-soluble components and dyes from the above-mentioned extract with n-hexane, petroleum ether or the like, and partitioning a solvent such as n-butanol, methyl ethyl ketone and water from the extract with water. To recover the proanthocyanidin into the solvent phase. As the column chromatography method, a method using normal-phase silica gel, a method using reverse-phase silica gel, Diaion HP-20 as a carrier, an adsorption column chromatography method using Sepabeads SP-207, and the like, Sephadex LH- as a carrier Two
A gel filtration method using 0 or the like can be mentioned, and these can be used alone or in combination and used repeatedly. Examples of the preparative high performance liquid chromatography method include a method using a reverse phase column using octadecyl silica or the like, a method using a normal phase column using silica gel or the like, and the like.

By the above-mentioned purification method, proanthocyanidins can be purified by removing water-soluble ionic substances such as salts, nonionic substances such as saccharides and polysaccharides, oils, pigments, and the like from the extract. In addition, grape-derived proanthocyanidins are available from Acta dermato
Dermato Venereologica), 78 , 428-432 (1998) or according to the method, procyanidin B-1 [epicatechin- (4β → 8) -catechin], procyanidin B-2.
[Epicatechin- (4β → 8) -epicatechin], procyanidin B-3 [catechin- (4α → 8) -catechin], procyanidin C-1 [epicatechin- (4β → 8) -epicatechin- (4β
→ 8) -Epicatechin] and procyanidin C-2 [catechin- (4α → 8) -catechin- (4α → 8) -catechin] are known from The Journal of Investigative Dermatology, 112. ,
Extraction and purification can be performed according to the method described in 310-316 (1999) or according to the method.

The production of proanthocyanidins by chemical synthesis is described in Journal of Chemical Parkin Transaction I (Journal of Chemi), which describes a method for producing a dimer of epicatechin or catechin.
cal Society, Perkin Transaction I), 1535-1543 (198
3) or phytochemistry, 25 ,
1209-1215 (1986) or a method analogous thereto.

When proanthocyanidins are used as the active ingredient of the present invention, they may be used alone or in combination of two or more. Specific examples thereof include proanthocyanidins derived from grape seeds, proanthocyanidins derived from apples, and barley derived from barley. Proanthocyanidins, pine-derived proanthocyanidins, purified procyanidins oligomers, procyanidins B-1, procyanidins B-2, procyanidins B-3, procyanidins C-1, one or more selected from procyanidins C-2 and the like, among which Procyanidin B-1, Procyanidin B-2, Procyanidin
One or more selected from B-3, procyanidin C-1 and procyanidin C-2 are preferably used.

As the tocopherol or tocopherol derivative used in the present invention, for example, any of commercially available natural and synthetic products can be used, and derivatives such as acetate and nicotinate can also be used. . Specific examples include dl-α-tocopherol, d-α-tocopherol, dl-α-tocopherol acetate, d-α-tocopherol acetate, dl-α-tocopherol nicotinate and the like.

As the pantothenic acid or pantothenic acid derivative used in the present invention, for example, any of commercially available natural and synthetic products can be used, and any pantothenic acid or pantothenic acid derivative can be used. Well, specifically, calcium pantothenate, sodium pantothenate, D-pantothenyl alcohol, DL-
Pantothenyl alcohol, pantothenyl ethyl ether and the like can be mentioned.

Protein kinase C used in the present invention
Any specific inhibitor can be used as long as it specifically inhibits protein kinase C. Preferably, protein kinase C (PKC) inhibitory activity and protein kinase A (PKA) inhibitory activity are as follows. PK to show
50% inhibition constant of PKA (hereinafter abbreviated as PKA-IC ₅₀ ) and PKC as measured by C inhibitory activity assay and PKA inhibitory activity assay
Of 50% inhibition constant (hereinafter abbreviated as PKC-IC ₅₀ )
PKA-IC ₅₀ / PKC-IC ₅₀ ) is 3 or more, preferably 3 to 1
0 ^9, protein kinase inhibitor is used more preferably 10 to 10 ^9. Specifically, calphosphin C,
One or more selected from hexadecylphosphocholine, palmitoyl-DL-carnitine, and polymyxin B or a pharmaceutically acceptable salt thereof can be given.

These pharmacologically acceptable salts include hydrochloride, hydrobromide, sulfate, nitrate, formate,
Acetate, benzoate, maleate, fumarate, succinate, tartrate, citrate, oxalate, methanesulfonate, toluenesulfonate, aspartate, glutamate, etc. . The method for measuring PKC inhibitory activity and the method for measuring PKA inhibitory activity are described below.

(1) Assay for PKC Inhibitory Activity PKC inhibitory activity was measured by the method of Yoshikawa et al. [Journal of Biologi
cal Chemistry), 257 , 13341 (1982)].

2.5 μmol magnesium acetate, 50 μg histone type IIIS (manufactured by Sigma), 20 μg phosphatidylserine, 0.8 μg diolein, 25 nmol calcium chloride, 5 nm
μg crude enzyme (partially purified from rat brain by the method of Yoshikawa et al.) and 5 μmol Tris-HCl buffer (pH 7.5)
The above aqueous solution containing a sample in a 250 μl aqueous solution containing (10 μl)
And incubate at 30 ° C for 3 minutes. After incubation, 1.25 nmol [γ- ³² P] ATP (5 to 10 × 10 ³ cpm / nmo
l) is added, phosphorylation is performed at 30 ° C. for 3 minutes, and the reaction is stopped by adding 25% trichloroacetic acid. The reaction solution was treated with a cellulose acetate membrane (pore size 0.45 μm) (manufactured by Toyo Roshi Kaisha, Ltd.).
After washing with 5% trichloroacetic acid four times, the radioactivity remaining on the membrane is measured to obtain a sample value. The above operation is performed without adding a sample, and the radioactivity is measured and used as a control value. The molar concentration of the sample showing 50% of the sample value with respect to the control value is defined as the PKC 50% inhibition constant (PKC-IC ₅₀ ).

(2) Method for Measuring PKA Inhibitory Activity PKA inhibitory activity can be measured according to the method of Kuo et al. [Biochemistry, 64 , 1349 (1969)].

5 μmol Tris-HCl buffer (pH 6.8), 2.5 μmol
The sample was placed in a 250 μl aqueous solution containing 100 μg magnesium acetate, 100 μg histone type IIS (manufactured by Sigma), 0.25 nmol c-AMP and 200 μg crude enzyme (partially purified from calf heart by the method of Kuo et al.). Containing the aqueous solution (10
μl) and incubate at 30 ° C. for 3 minutes. After incubation, 1.25 nmol [γ- ³² P] ATP (5 to 10 × 10 ³ cpm / n
mol), phosphorylation reaction is performed at 30 ° C. for 3 minutes, and the reaction is stopped by adding 25% trichloroacetic acid. The reaction solution is filtered through a cellulose acetate membrane (pore size 0.45 μm) (manufactured by Toyo Roshi Kaisha, Ltd.), washed four times with 5% trichloroacetic acid, and the radioactivity remaining on the membrane is measured to obtain a sample value. The above operation is performed without adding a sample, and the radioactivity is measured and used as a control value.

The molar concentration of the sample when the sample value shows 50% of the control value is defined as the 50% inhibition constant of PKA (PKA-IC ₅₀ ). As biotin used in the present invention, for example, any of commercially available natural products and synthetic products can be used, and D-biotin is preferable. The dosage form of the hair restorer of the present invention includes phosphatidic acid represented by the formula (I); said phosphatidic acid, proanthocyanidin, tocopherol, tocopherol derivative, pantothenic acid, pantothenic acid derivative, protein kinase C-specific inhibitor or A pharmacologically acceptable salt thereof, and one or more components selected from the group consisting of biotin; phosphatidic acid;
One or more components selected from the group consisting of proanthocyanidins, tocopherols, tocopherol derivatives, pantothenic acid, pantothenic acid derivatives and protein kinase C-specific inhibitors or pharmacologically acceptable salts thereof;
Or phosphatidic acid and biotin, and one or more components selected from the group consisting of proanthocyanidins, tocopherols, tocopherol derivatives, pantothenic acid, pantothenic acid derivatives and protein kinase C-specific inhibitors or pharmacologically acceptable salts thereof. Any dosage form can be used as long as the dosage form can be blended with For example, it can be used as a liquid or solid hair restorer by blending with an appropriate pharmaceutical base.

Examples of the liquid or solid hair growth preparation include liquid preparations such as hair liquid, hair tonic, and hair lotion, and solid preparations such as ointment and hair cream. A phosphatidic acid represented by I): from the phosphatidic acid and proanthocyanidin, tocopherol, a tocopherol derivative, pantothenic acid, a pantothenic acid derivative, a protein kinase C-specific inhibitor or a pharmaceutically acceptable salt thereof, and biotin. At least one component selected from the group consisting of: phosphatidic acid, proanthocyanidin, tocopherol, a tocopherol derivative, pantothenic acid, a pantothenic acid derivative, and a protein kinase C-specific inhibitor or a pharmaceutically acceptable salt thereof. One or more components selected from; or Sphatidic acid and biotin, and one or more components selected from the group consisting of proanthocyanidins, tocopherols, tocopherol derivatives, pantothenic acid, pantothenic acid derivatives and protein kinase C-specific inhibitors or pharmacologically acceptable salts thereof. And then can be produced by a conventional method.

The content of phosphatidic acid in the hair restorer of the present invention varies greatly depending on the type of phosphatidic acid and percutaneous absorption derived from physical properties, but is usually 0.01 to 5.0% by weight alone (hereinafter simply referred to as a mixture). %), Preferably 0.01 to 3.0%, more preferably 0.1 to 1.0%. The content of proanthocyanidins varies depending on the degree of purification, but is usually 0.01 to 10.0%, preferably 0.1 to 5.0%,
More preferably, it is 0.3 to 2.0%. The content of tocopherol or tocopherol derivative is usually 0.01 to 2%,
Preferably it is 0.05-2%, more preferably 0.05-1%. The content of pantothenic acid or a pantothenic acid derivative is usually 0.01 to 2%, preferably 0.05 to 1%, more preferably 0.1 to 0.5%. The content of the protein kinase C inhibitor or a pharmacologically acceptable salt thereof greatly varies depending on the strength of the inhibitory activity and transdermal absorbability derived from physical properties, but is usually 0.00001 to 1% alone or as a mixture, preferably Is 0.0001 to 1%, more preferably 0.001 to 0.1%. The content of biotin is usually 0.0001 to 0.1%, preferably 0.001 to 0.1%, more preferably 0.001 to 0.05%.

Suitable bases for the liquid dosage form include those usually used in hair restorers, for example, purified water, ethyl alcohol, polyhydric alcohols and the like, and additives may be added as necessary. . Examples of the polyhydric alcohol include glycerol, 1,3-butylene glycol, propylene glycol and the like.

As additives, surfactants, vitamins, anti-inflammatory agents, bactericides, hormonal agents, crude drug extracts, tinctures, cooling agents, humectants, antioxidants, sequestering agents,
Perfumes and the like. As surfactants, polyoxyethylene (60) hydrogenated castor oil, polyoxyethylene (8) oleyl ether, polyoxyethylene (10) oleyl ether, polyoxyethylene monooleate (10), polyoxyethylene (30) Glyceryl monostearate, sorbitan monostearate, polyoxyethylene (20) sorbitan monooleate, sucrose fatty acid ester, hexaglycerin monooleate, hexaglycerin monolaurate, polyoxyethylene reduced lanolin, polyoxyethylene (20) lanolin alcohol,
Polyoxyethylene (25) glyceryl pyroglutamic acid isostearic acid diester, N-acetylglutamine isostearyl ester and the like can be mentioned.

The vitamins include benzyl nicotinate, nicotinamide, pyridoxine hydrochloride, riboflavin and the like. Anti-inflammatory agents include dipotassium glycyrrhizinate, β-glycyrrhetinic acid, allantoin, diphenhydramine hydrochloride, guaiazulene, l-menthol and the like.

Examples of fungicides include trichlorohydroxydiphenyl ether, hinokitiol, triclosan, chlorhexidine gluconate, phenoxyethanol,
Resorcinol, isopropylmethylphenol, azulene, salicylic acid, zinc pyrithione, benzalkonium chloride, photosensitive element 301, sodium mononitroguaiacol, and the like.

Examples of the hormonal agent include ethinyl estradiol, estrone, estradiol and the like. Examples of the crude drug extracts include the assembly extract, garlic extract, carrot extract, aloe extract, kina extract, cordyceps extract, saffron extract and the like. As tinctures, pepper tincture, ginger tincture, cantaris tincture and the like can be mentioned.

[0060] Examples of the cooling agent include pepper tincture, l-menthol, camphor and the like. As a moisturizer,
L-pyrrolidone carboxylic acid, sodium hyaluronate, chondroitin sulfate and the like. Examples of the antioxidant include butylhydroxyanisole, isopropyl gallate, propyl gallate, erythorbic acid, and the like.

Examples of the sequestering agent include ethylenediaminetetraacetate or a salt thereof. As fragrance, orange oil, lemon oil, bergamot oil,
Natural flavors such as lime oil, lemongrass oil and lavender oil and synthetic flavors such as menthol, rose oxide, linalool, citral, linalyl acetate and the like can be mentioned.

When the above liquid dosage form is used as a spray, a combustible gas, a non-combustible gas or the like can be used. Examples of the combustible gas include LPG (liquefied petroleum gas) and dimethyl ether, and examples of the non-combustible gas include nitrogen gas and carbon dioxide gas. Examples of the solid dosage form base include vaseline, solid paraffin, vegetable oil, mineral oil, lanolin, waxes, macrogol, and the like.If necessary, the above additives, ethyl alcohol, lower alcohols such as isopropyl alcohol, etc. It may be added.

The dosage of the hair restorer of the present invention varies depending on age, body weight, symptoms, therapeutic effect, administration method, treatment time and the like.
0.1 to 2 phosphatidic acids at a time per adult
50 mg, preferably 1 mg to 100 mg, is transdermally administered once to several times a day. Next, the present invention will be described in detail with reference to examples.

[0064]

EXAMPLES Example 1 Preparation of Composition 1 and Composition 2 1-O-hexadecyl-2-O-methylglyceryl-3-phosphate 0.4% ethyl alcohol 70% 1,3-butylene glycol 3% N- Acetylglutamine isostearyl ester 0.25% Polyoxyethylene (25) glyceryl pyroglutamate isostearic acid ester 0.25% Purified water was added to the above mixture to make 100%, and this was homogenized while stirring to prepare composition 1. In the above composition, purified water was added in place of 1-O-hexadecyl-2-O-methylglyceryl-3-phosphate to prepare composition 2.

Example 2: Preparation of Composition 3 and Composition 4 1-O-hexadecyl-2-O-methylglyceryl-3-phosphate 0.4% Procyanidin B-2 1% Ethyl alcohol 70% 1,3-butylene Glycol 3% N-acetylglutamine isostearyl ester 0.25% Polyoxyethylene (25) glyceryl pyroglutamate isostearic acid ester 0.25% Procyanidin B-2 is a product of The Journal of Investigative Dermatology.
gative Dermatology), 112 , 310-316 (1999). Add purified water to the above mixture and add 100
%, And the mixture was made uniform with stirring to prepare Composition 3. In the above composition, 1-O-hexadecyl-2-O-
Purified water was added in place of methylglyceryl-3-phosphate to prepare composition 4.

Example 3 Preparation of Composition 5 and Composition 6 1-O-hexadecyl-2-O-methylglyceryl-3-phosphate 0.4% dl-α-tocopherol 1.0% Ethyl alcohol 70% 1,3- Butylene glycol 3% N-acetylglutamine isostearyl ester 0.25% Polyoxyethylene (25) glyceryl pyroglutamate isostearic acid ester 0.25% Purified water is added to the above mixture to make 100%, and the mixture is stirred to make it uniform, and the composition is adjusted. Preparation 5 was prepared. In the above composition, purified water was added in place of 1-O-hexadecyl-2-O-methylglyceryl-3-phosphate to prepare composition 6.

Example 4: Preparation of Composition 7 and Composition 8 1-O-hexadecyl-2-O-methylglyceryl-3-phosphoric acid 0.4% Pantothenyl ethyl ether 0.3% Ethyl alcohol 70% 1,3-butylene Glycol 3% N-acetylglutamine isostearyl ester 0.25% Polyoxyethylene (25) glyceryl pyroglutamate isostearic acid ester 0.25% Purified water is added to the above mixture to make 100%, and the mixture is stirred to make it uniform, and the composition is made up. 7 was prepared. In the above composition, purified water was added in place of 1-O-hexadecyl-2-O-methylglyceryl-3-phosphate to prepare composition 8.

Example 5 Preparation of Composition 9 and Composition 10 1-O-hexadecyl-2-O-methylglyceryl-3-phosphate 0.4% biotin 0.05% ethyl alcohol 70% 1,3-butylene glycol 3% N-acetylglutamine isostearyl ester 0.25% Polyoxyethylene (25) glyceryl pyroglutamic acid isostearate ester 0.25% Purified water is added to the above mixture to make 100%, and the mixture is homogenized while stirring to prepare composition 9. did. In the above composition, purified water was added in place of 1-O-hexadecyl-2-O-methylglyceryl-3-phosphate to prepare composition 10.

Example 6 Preparation of Composition 11 1-O-oleoyl-2-O-methylglyceryl-3-phosphate 0.4% ethyl alcohol 70% 1,3-butylene glycol 3% N-acetylglutamine isostearyl Ester 0.25% Polyoxyethylene (25) glyceryl pyroglutamate isostearic acid ester 0.25% Purified water was added to the above mixture to make 100%, and this was homogenized while stirring to prepare composition 11.

Example 7: Preparation of composition 12 1-O-hexadecyl-2-O-acetylglyceryl-3-phosphate 0.4% ethyl alcohol 70% 1,3-butylene glycol 3% N-acetylglutamine isostearyl ester 0.25% Polyoxyethylene (25) glyceryl pyroglutamate isostearic acid ester 0.25% Purified water was added to the above mixture to make 100%, and the mixture was homogenized with stirring to prepare Composition 12.

Reference Example 1: Synthesis of 1-O-hexadecyl-2-O-methylglyceryl-3-phosphate (1-O-hexadecyl-2-O-methylphosphatidic acid) Journal of Medicinal Chemistry (Journal)
of Medicinal Chemistry), 2038-2044 (1986) or a method analogous thereto. 1-O-
Hexadecyl-2-O-methylglycerol (Sigma) 165
was dissolved in 10 mL of hexane, and 700 μl of phosphorus oxychloride, 1000 μl of triethylamine and
The mixture was added dropwise to the mixed solution at room temperature with stirring. 15
After stirring for an hour, purified water (10 ml) was added, and the mixture was further stirred at room temperature for 1 hour. The organic layer was obtained and developed with a mobile phase of chloroform: methanol: acetic acid: water = 170: 25: 25: 6 using a silica gel thin-layer chromatography plate, and the band containing the target substance was scraped off. Diethyl ether was added to the silica gel powder to obtain an organic layer.
The organic layer was further concentrated under reduced pressure to give 1-O-hexadecyl-2-O
-Methylglyceryl-3-phosphate was obtained in an amount of 52 mg.

Reference Example 2: Synthesis of 1-O-oleoyl-2-O-methylglyceryl-3-phosphate (1-O-oleoyl-2-O-methylphosphatidic acid) Annual New York Academic Society (Ann. NY Acad. Sci), 279-281 (2000) or WO
The method can be carried out by the method described in 99/47101 or a method analogous thereto. Monoolein (Kishida) 1020
Using mg as a starting material, the alcohol residue at position 3 was protected with triphenylmethyl group with triphenylchloromethane, and then methyl iodide and sodium hydride were reacted to introduce a methyl group into the alcohol residue at position 2. . Further, the resulting 1-O-oleoyl-2-O-methyl-3-O-triphenylmethylglycerol was treated with acetic acid to deprotect the triphenylmethyl group, and the resulting 1-O-oleoyl- 2-O
-Methylglycerol is reacted with phosphorus oxychloride and triethylamine to give the desired 1-O-oleoyl-2-O
271 mg of -methylglyceryl-3-phosphoric acid were obtained.

Reference Example 3: Synthesis of 1-O-hexadecyl-2-O-acetylglyceryl-3-phosphate (1-O-hexadecyl-2-O-acetylphosphatidic acid) Journal of Medicinal Chemistry (Journal)
of Medicinal Chemistry), 2038-2044 (1986) or a method analogous thereto. 1-
179 mg of O-hexadecyl-2-O-acetylglycerol (Funakoshi) was dissolved in 17.9 ml of hexane, and 700 μl of phosphorus oxychloride, 1000 μl of triethylamine and hexane 5
The mixture was added dropwise to 0 ml of the mixed solution at room temperature with stirring. 15
After stirring for an hour, purified water (10 ml) was added, and the mixture was further stirred at room temperature for 1 hour. The organic layer was obtained and developed using a silica gel thin-layer chromatography plate with a mobile phase of chloroform: methanol: acetic acid: water = 170: 25: 25: 6, and the band containing the target substance was scraped off. Diethyl ether was added to the silica gel powder and mixed well to obtain an organic layer. The organic layer was further concentrated under reduced pressure to give 1-O-
Hexadecyl-2-O-acetylglyceryl-3-phosphate 42 mg
I got

Next, the action of the hair restorer of the present invention will be specifically shown by test examples. Test Example 1: Promoting effect on mouse hair follicle epithelial cell culture Hair follicle epithelial cells were separated and cultured according to the method of Tanigaki et al. [Archives of Dermatological Research (Archive).
s of Dermatological Research), 284 , 290-296 (199
2)] was modified.

More specifically, the back skin of a 4-day-old C3H mouse (Charles River Japan) was collected, and a MEM medium (Eagle's Minimum) containing 500 units / ml of dispase (Kodoshu) and 5% fetal calf serum (FCS) was collected. Essential Medium) 4
The treatment was performed at 16 ° C. for 16 hours. The epidermis was peeled off from the obtained skin slice, and the dermis layer was placed in a DMEM medium (Dulbecco's modified Eag) containing 0.25% collagenase N-2 (Nitta gelatin) and 10% FCS.
le Medium) at 37 ° C for 1 hour to obtain a dermal suspension. After filtering the dermis suspension through a 212 micron nylon mesh (Nippon Riken Co., Ltd.), the filtrate was centrifuged at 1000 rpm for 5 minutes to obtain a pellet containing hair follicle tissue. Calcium / magnesium free PBS (Dulbecco's Phosphat)
e-Buffered Saline) solution was added thereto, suspended using a pipette, and allowed to stand for 15 minutes to sediment the hair follicle tissue. Using the obtained hair follicle tissue, performed on the above pellets, addition of a calcium magnesium free PBS solution,
The same operation as suspension with a pipette and standing / sedimentation for 15 minutes was repeated three times.

A 0.1% ethylenediaminetetraacetic acid (EDTA) -0.25% trypsin solution (manufactured by Gibco) was applied to the obtained hair follicle tissue.
After treatment at 37 ° C. for 5 minutes, a DMEM medium containing 10% FCS was added to prepare a hair follicle tissue cell solution having a cell concentration of 3 × 10 ⁵ / ml. The hair follicle tissue cell solution was seeded at 1 ml / well on a 24-well collagen coated plate (manufactured by Iwaki Glass Co., Ltd.),
Culture was performed for 24 hours under 5% CO ₂ .

After the culture, bovine insulin (manufactured by Far East Pharmaceutical) 5 mg / L; mouse epidermal growth factor (EGF) (manufactured by Takara Shuzo) 5 μg / L in MCDB153 medium (manufactured by Far East Pharmaceutical Co.); 40mg / L; human transferrin (Sigma) 10mg / L; hydrocortisone (Sigma)
0.4 mg / L; progesterone (manufactured by Collaborative Research) 0.63 μg / L; O-phosphoethanolamine (manufactured by Sigma) 14 mg / L; ethanolamine (manufactured by Sigma) 6.1 mg / L
L; penicillin (manufactured by Wako) 50 U / ml; streptomycin (manufactured by Wako) 50 μg / ml; 1-O-hexadecyl-2-O-methylglyceryl-3-phosphate, 1-O-oleoyl-2-O DMSO solution containing phosphatidic acid such as 1-methylglyceryl-3-phosphate, 1-O-hexadecyl-2-O-acetylglyceryl-3-phosphate, and / or procyanidin B-2 or dl-α-tocopherol (1 The medium was replaced with a medium containing (/ 100 volumes added), and the cells were further cultured at 37 ° C in the presence of 5% CO ₂ for 5 days. The medium was replaced every other day.

In the above medium, 1/10 of DMSO alone was used instead of the DMSO solution containing phosphatidic acid and / or procyanidin B-2 or dl-α-tocopherol.
A culture cultured in a medium to which 0 volume was added was used as a control group. Measurement of cell proliferation was performed using MTT [3- (4,5-Dimethylthiazol-2-y
l) -2,5-diphenyltetrazolium bromide] [Experimental Medicine Separate Volume Bio Manual UP Series Cultured Cell Experiments for Molecular Biology Research, pp. 89-92,
Youtosha (1995)].

In each well of a 24-well microplate (2 cm ² / well), 1/10 volume of a PBS solution of MTT (5
mg / ml), shaken to homogenize, and cultured at 37 ° C in the presence of 5% CO ₂ for 4 hours. After 4 hours, the culture solution was aspirated, 1 ml of 0.04 mol / L HCl isopropyl alcohol solution was added to each plate, and mixed until the formazan formed in the well was completely dissolved.

Using 650 nm as a control, the absorbance at 570 nm was measured to determine the degree of cell proliferation. Table 1 shows the growth promoting activity of the compounds of the present invention.

[0081]

[Table 1]

As shown in Table 1, the phosphatidic acid of the present invention exhibited a remarkable mouse hair follicle epithelial cell growth promoting activity. In addition, the activity of proanthocyanidins and tocopherols to promote mouse hair follicle epithelial cell proliferation was enhanced by mixing with the above-mentioned phosphatidic acid.

Test Example 2: Effect on Hair Growth in Mice Ogawa et al. [Journal of Dermatology (The J
Ournal of Dermatology), 10 , 45-54 (1983)], a mouse hair growth effect test was conducted. 9-week-old C3H / HeSlc male mice in the telogen phase of the hair cycle (groups 4 to 5)
The back hair of each of the animals was shaved with an electric clipper and an electric shaver, and the compositions 1 to 12 prepared in Examples 1 to 7 were uniformly applied to the shaved portion once a day, 200 μl each. In addition, the composition 2 was set as the control group. The skin on the back of the mouse 18 days after the start of the test application was collected and photographed, and then the percentage of the hair growth portion relative to the total area of the back skin was determined using an image analysis processor (Avionics, Spica II). The value obtained by subtracting the value of the hair growth rate of the control group from the value of the hair growth rate of the test drug group was defined as an increased hair growth area rate (%).

The results are shown in Table 2.

[0085]

[Table 2]

As shown in Table 2, the hair restorer containing phosphatidic acid of the present invention showed a remarkable mouse hair growth promoting effect. The hair growth promoting effects of proanthocyanidins, tocopherols, pantothenic acid derivatives and biotin were enhanced by mixing with phosphatidic acid.

[0087]

According to the present invention, a hair restorer excellent in hair growth and hair growth effects of hair characterized by containing a phosphatidic acid having a specific chemical structure, and the above phosphatidic acid and proanthocyanidin , Tocopherols,
Tocopherol derivatives, pantothenic acid, pantothenic acid derivatives, protein kinase C-specific inhibitors or pharmacologically acceptable salts thereof, and containing one or more components selected from the group consisting of biotin as an active ingredient. It is possible to provide a hair restorer or the like which is characterized.

 ────────────────────────────────────────────────── ─── Continuing from the front page (72) Inventor Shinkichi Honda 1-6-1, Otemachi, Chiyoda-ku, Tokyo Kyowa Hakko Kogyo Co., Ltd. F-term (reference) 4C083 AC102 AC122 AC641 AC662 AC841 AC842 AC851 AC852 AC901 AC902 AD611 AD631 AD661 AD662 CC37 DD23 EE22

Claims (19)

[Claims] (1) Formula (I) (Wherein, R ¹ represents alkyl, alkenyl, alkanoyl or alkenoyl; R ¹ represents alkyl or alkenyl; R ² represents alkyl, alkenyl, alkanoyl or alkenoyl; R ¹ represents alkanoyl or alkenoyl; R ² represents alkyl or alkenyl) as an active ingredient. 2. R ¹ is alkyl or alkenyl,
The hair restorer according to claim 1, wherein R ² is methyl. 3. The hair restorer according to claim 1, wherein R ¹ is alkanoyl or alkenoyl, and R ² is methyl. 4. R ¹ is alkyl or alkenyl,
The hair restorer according to claim 1, wherein R ² is acetyl. 5. The hair restorer according to claim 1, wherein R ¹ is octadecenoyl or hexadecyl, and R ² is methyl. 6. The hair restorer according to claim 1, wherein R ¹ is hexadecyl and R ² is acetyl. 7. The phosphatidic acid according to any one of claims 1 to 6, and proanthocyanidin, tocopherol, a tocopherol derivative, pantothenic acid, a pantothenic acid derivative, a protein kinase C-specific inhibitor or a pharmacologically acceptable thereof. A hair restorer comprising, as an active ingredient, at least one ingredient selected from the group consisting of salt and biotin. 8. The hair restorer according to claim 1, further comprising proanthocyanidin. 9. The proanthocyanidin is procyanidin.
The hair restorer according to claim 8, which is one or more proanthocyanidins selected from the group consisting of B-1, procyanidin B-2, procyanidin B-3, procyanidin C-1 and procyanidin C-2. 10. The hair restorer according to claim 1, which contains tocopherol or a tocopherol derivative. 11. A tocopherol or a tocopherol derivative selected from the group consisting of dl-α-tocopherol, d-α-tocopherol, dl-α-tocopherol acetate, d-α-tocopherol acetate and dl-α-tocopherol nicotinate The hair restorer according to claim 10, which is a tocopherol or two or more tocopherol derivatives. 12. The method according to claim 1, further comprising pantothenic acid or a pantothenic acid derivative.
The hair restorer according to any one of 11 above. 13. One or more pantothenic acids wherein the pantothenic acid or pantothenic acid derivative is selected from the group consisting of calcium pantothenate, sodium pantothenate, D-pantothenyl alcohol, DL-pantothenyl alcohol and pantothenyl ethyl ether 13. The hair restorer according to claim 12, which is a pantothenic acid derivative. 14. The hair restorer according to any one of claims 1 to 6 or 8 to 13, comprising a protein kinase C-specific inhibitor or a pharmacologically acceptable salt thereof. 15. The protein kinase C-specific inhibitor is one or more protein kinase C inhibitors selected from calphosphin C, hexadecylphosphocholine, palmitoyl-DL-carnitine and polymyxin B. Hair restorer. 16. The hair restorer according to any one of claims 1 to 6 or 8 to 15, which comprises biotin. 17. The hair restorer according to claim 1, which is substantially free of minoxidil. 18. A phosphatidic acid and one or more selected from the group consisting of proanthocyanidins, tocopherols, tocopherol derivatives, pantothenic acid, pantothenic acid derivatives, and protein kinase C-specific inhibitors or pharmacologically acceptable salts thereof. A hair restorer containing the following components as active ingredients. 19. A phosphatidic acid and biotin,
Proanthocyanidins, tocopherols, tocopherol derivatives, pantothenic acid, containing one or more components selected from the group consisting of pantothenic acid derivatives and protein kinase C-specific inhibitors or pharmacologically acceptable salts thereof as an active ingredient Hair restorer.