JPH10279501A - Hair growing agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a hair growing agent developing in a hair papilla cell, providing a paracrine factor to be a key to control hair growth, having growth promoting action on hair follicle, by including a fibroblast growth factor FGF-10 as an active ingredient. SOLUTION: This hair growing agent comprises a fibroblast growth factor FGF-10 being essential to organisms and high in safety as an active ingredient, having hair growing effects, treating effects on depilation, preventing effects on depilation, etc., useful for a preventive and a therapeutic agent for alopecia and is useful as a hair growing/nourishing cosmetic and a depilation preventing cosmetic.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a hair restorer containing FGF-10, which belongs to the fibroblast growth factor (FGF) family, as an active ingredient.

[0002]

2. Description of the Related Art Conventional hair restorers use a "cell activating" or "activating" action as Merkmal to promote the metabolism of tissues around hair follicles and to increase the blood flow in the capillaries of tissues surrounding hair follicles. Development has centered on promoting the proliferation of epithelial cells (such as hair matrix cells). As a result, placenta extract, carrot extract, assembly extract, pantothenic acid and its derivatives, vitamin E and its derivatives, pepper tincture, nicotinic acid derivatives, and the like have been used in hair restorers as active ingredients having cell activating and blood circulation promoting effects. Was.

However, versatile cell activators and activators, which are only expected to activate the hair follicle epithelial cells, which are the constituent cells of hair, are frequently administered in large quantities due to the limitation of the amount of the compound. Inevitably results in many drawbacks (such as irritation).

By the way, in the hair follicle tissue involved in hair growth, especially in the lower part, undifferentiated hair mother cells in the epithelium are present so as to surround the mesenchymal hair papilla cells and the basement membrane, It is thought that normal hair growth is caused by the proliferation and differentiation of hair cells by these tissue interactions. That is, the hair matrix controlled by the hair papilla cells proliferates and differentiates, and the hair pulp, the cortex, the hair shaft of the hair dermal and the like, which forms the main body of the hair, and the inner root sheath are formed. It is thought that hair elongates when hair mother cells move upward due to proliferation and differentiation.

For example, when hair papilla cells and cultured dermal papilla cells are implanted between epidermis and dermis of mouse or rat plantar skin without hair follicles and transferred into the transplantation bed, the plantar epidermis which does not normally induce hair follicles becomes It has been reported that hair papilla cells are enveloped and induce hair follicles (Kobayashi, K., et al., J. Invest. Dermato
l., 92, p278-282 (1989), Reynolds, A., et al., Ann.NYAca
d.Aci., 642, p226-242 (1991)). It has also been reported that the type and size of the embedded nipple regulates the induced hair follicle (Jahoda, C., Development, 115, p1103-1109).
(1992)).

On the other hand, regarding the interaction between hair follicle tissues,
There are reports on androgen-dependent hair growth. For example, the proliferation of beard epithelial cells in the presence of androgen-dependent papillary cells is dependent on insulin-like growth factor-I (insuli
It is completely suppressed by neutralizing antibody against n-like growth factor-I (IGF-I), and IGF-I
MRNA is expressed, and since this expression is controlled by male hormones, at least IGF-
It has been shown that I is one of the growth factors that act directly on neighboring cells by secretion (paracrine) in the interaction between androgen-dependent hair follicle tissues derived from dermal papilla cells (Itami, S., et al., Biochem. Biophys. Res. Co.
mmun.212, p988-994 (1995), Itami, S., et al., Hair Rese
ach for the Next Millenium p297-302 (1996) Elsevier
Science).

At present, it is certain that hair papilla cells, which are mesenchymal cells, are the key to hair growth, but there are still many unclear points about the mechanism of interaction between hair follicle tissues. The elucidation is awaited.

[0008]

Accordingly, the present invention provides a paracrine factor which can be involved in the interaction between hair follicle tissues, in particular, a paracrine factor which is expressed in dermal papilla cells which is a key to hair growth. The task is to provide. Thus, an object of the present invention is to provide a hair restorer containing the factor as an active ingredient.

[0009]

Means for Solving the Problems Hair papilla plays an important role not only in inducing differentiation from skin tissue into hair follicle tissue, but also in maintaining the hair cycle and in differentiation and proliferation of hair matrix cells during anagen. It is a cell that is thought to be. It is thought that signals emitted from the hair papilla cells are deeply involved in the role of these hair papilla cells. Therefore, it can be said that determining this signal is very important in elucidating the mechanism of interaction between hair follicle tissues.

[0010] The present inventors have conducted a sincere study to solve the above-mentioned problems, and as a result, recently found a fibroblast growth factor-10 (FGF-10) (Yamasaki, M., et al. , J. Biol. Chem.
271, p15918-15921 (1996), Emoto et al., The 69th Annual Meeting of the Biochemical Society of Japan, 19th Annual Meeting of the Molecular Biology Society of Japan, 163 (1996)) It was revealed that FGF-10 was expressed in hair papilla cells and had a hair follicle growth effect. Thus, the present inventors have found that FGF-10 can be used as a hair restorer,
The present invention has been completed.

FGF-10 belongs to the FGF family isolated from a fetal rat cDNA library by PCR using a part of a region retaining an amino acid sequence highly homologous to the FGF family as a primer. The tenth factor (Yamasaki, M., et al., J. Biol. Chem. 271, p15918-15
921 (1996)). Regarding members belonging to the FGF family, they have been used as melanin synthesis inhibitors (Japanese
No. 7-304686), use as a liver function improving agent
No. 6-345666) and its use as a skin cosmetic (Japanese Patent Application Laid-Open No. 4-187613). Use for treatment and prevention of baldness (Japanese Patent Application Laid-Open No. H4-224522)
And use as hair nourishing cosmetics and gray hair improving cosmetics
No. 8-208440). Meanwhile, FG
F-10 has been reported in rats. In embryos, posterior pituitary, first cervical vertebrae (fir
st cervical vertebra), duodenum (duodenum), lung (l
ung), sacral and coccyg
eal segments of the spinal cord), specifically expressed in the lung in adults (Yamasaki, M., et al., J. Biol. Chem. 271, p.
15918-15921 (1996)). It has also been reported that it has an epithelial cell proliferation activity. However, the relationship between FGF-10 and human papillary cells, which are responsible for the growth of human hair tissue, particularly hair growth, has not been known at all.

The present inventors isolated hair papilla cells, epidermal keratinocytes of the epithelial system, and outer root sheath cells, and performed RT-PCR on total RNA in these cells. For the first time, it was revealed that it is expressed in hair papilla cells, which play a central role in hair growth. Furthermore, the present inventors evaluated hair follicle growth by applying FGF-10 to hair follicle tissue prepared from isolated hair follicles, and FGF-10 actually has a hair follicle growth promoting action. That was revealed for the first time.

The present invention relates to a paracrine factor "FGF-10" having a hair follicle growth promoting action, and more specifically to FGF-10.
And a hair restorer containing as an active ingredient.

[0014]

BEST MODE FOR CARRYING OUT THE INVENTION FGF-10 used as an active ingredient of the hair restorer of the present invention can be produced from a tissue or cell of a warm-blooded animal by a known protein purification method. It can also be manufactured using. Natural FGF-10 is, for example, commonly used chromatography techniques and salting out methods, solvent extraction methods, ultrafiltration concentration methods,
Purification and isolation can be achieved by appropriately combining recrystallization, centrifugation, distillation, and the like. When FGF-10 is produced using recombinant technology, the nucleotide sequence used to express FGF-10 may be the nucleotide sequence described in SEQ ID NO: 2 in addition to the nucleotide sequence described in SEQ ID NO: 1. Any nucleotide sequence can be used as long as it has a nucleotide sequence encoding a protein substantially identical to the amino acid sequence. The recombinant FGF-10 is, for example, a host cell is transformed with a recombinant expression vector containing a clone consisting of a nucleotide sequence encoding the entire amino acid sequence or partial amino acid sequence of FGF-10, and the resulting transformant is obtained. Is cultured in a suitable medium and under suitable culture conditions, and then extracted and purified from the transformant. The expression vector used is not particularly limited as long as it can be replicated, propagated, transcribed and translated in the host. As a host into which the vector is introduced, for example, Escherichia coli, yeast, insect cells, mammalian cells, and the like can be used.

In the present invention, the modified FGF-
It is also possible to use 10 polypeptides. An altered polypeptide may be naturally occurring or may be due to post-translational modification. In addition, genetic engineering techniques such as site-directed mutagenesis [Methods in Enzymol
ogy, 154, p350,367-382 (1987), 100, p468 (1983), Nuc
leic Acids Research, 12, p9441 (1984)], and chemical synthesis methods such as the phosphate triester method and the phosphate amidite method [J. Am. Chem. Soc., 89, p4801 (1967), 91]. , p3350 (1
969); Science, 150, p178 (1968), Tetrahedron Lett., 2
2, p1859 (1981), 24, p245 (1983)]
It can also be prepared by using A. Furthermore, it is also possible to prepare by appropriately combining these methods.

The FGF-10 of the present invention can be used alone as a hair restorer, but it can also be used in combination with other components. Other components include a cell activating effect, a blood circulation promoting effect, an anti-androgen, and the like. Specifically, pantothenic acid and its derivatives, placenta extract, photosensitizer, carrot extract, biotin, mononitroguaiacol, carpronium chloride, vitamin E and its derivatives, assembly extract, capsicum tincture, cepharanthin, nicotinic acid and its derivatives, estradiol , Ethinylestradiol, landic acid, minoxidil and its analogs and derivatives, 5α-reductase inhibitor, 12-tetradecanoylphorbol-13-acetate (1
Herbal medicines such as 2-Tetradecanoylphorbol-13-acetate), oak, oak, milk thistle, henna, and licorice, but are not limited thereto. These ingredients are 2
It is also possible to use combinations of more than one species. These formulations may be in any proportion that can effectively promote hair growth or prevent hair loss.

When the FGF-10 of the present invention is administered as a topical agent, it can be usually administered together with an acceptable carrier or vehicle. For example, cosmetics, pharmaceuticals, aqueous components used in quasi-drugs, powders,
Surfactants, oils, humectants, alcohol, pH adjusters, preservatives, antioxidants, thickeners, vitamins, sebum dissolving agents,
A bactericide, a keratolytic agent, a fragrance, a pigment, and the like can be added. In the case of topical administration, it can be used together with a known DDS carrier such as a liposome for more effective use.

The dosage of the hair restorer of the present invention is usually about 0.001.
~ 100μg / day / cm ² , 0.0001 ~ 0.1w / v%
And preferably from about 0.01 to about 10 μg / day / cm ² , 0.0005
~ 0.05w / v%. The daily dose is appropriately selected depending on the condition of the scalp and the like.
It can be administered in several divided doses.

The hair restorer of the present invention can be made into pharmaceutical dosage forms such as creams, lotions, ointments, gels, shampoos and aerosols, and cosmetic dosage forms.

Since the FGF-10 of the present invention is considered to have a hair-growth effect, and also to have a hair loss treatment effect and a hair loss prevention effect, the hair growth agent of the present invention can be used as a hair loss prevention agent or a hair loss treatment agent. It is also possible to use. Further, since it is derived from a living body and has high safety, it can be used as a hair growth / hair restoration cosmetic or a hair loss preventing cosmetic.

[0021]

【Example】

[Example 1] Isolation of dermal papilla cells From full-thickness skin obtained during plastic surgery, a conventional method (Me
ssenger, AG, Br. J. Dermatology., 110, p685-689 (198
Hair papilla cells were isolated according to 4)). The following operations were all performed under a stereo microscope in a clean bench. Hair follicles including peripheral tissues were selected and removed from the full-thickness skin using a 21G injection needle. From the excised hair follicle tissue, adhering peripheral tissue was surgically removed, and only the hair bulb was cut off. The hair bulb was incised, and only the dermal papilla was isolated and used for primary culture. Put 4 to 5 hair papilla obtained as described above per 35 mm culture dish, penicillin (100 U / ml), streptomycin (100 μg / ml) at 37 ° C., 5% CO ₂ , humidified conditions, Culture is performed using 199 medium (GIBCO BRL) containing glutamine (2 mmol / ml) and 20% fetal bovine serum, and the medium is replaced after the dermal papilla adheres to the bottom of the dish.
Performed every three days. In addition, epidermal keratinocytes are "Epi
dercell NHEK (F) ”(Kurabo Industries), and the cells were“ H
The culture was performed in "uMedia-KG2 medium" (Kurabo Industries, Ltd.). In addition, outer root sheath cells were isolated by a known method, and cultured in "MCDB153 medium" (Sigma). Subculture of any of the above cells was performed according to a conventional method using a PBS (−) solution containing 0.05% trypsin and 0.02% EDTA after subconfluence. In the experiment, cells cultured for 3 to 4 passages were used.

Example 2 RT-PCR (Reverse transcript)
ase-polymerase chain reaction) (1) Purification of total RNA Purification of total RNA from cultured dermal papilla cells can be performed according to a conventional method. Specifically, a guanidium thiocyanate-phenol-chloroform method (Chomczynski,
P., et.al., Anal. Biochem., 162, p156-159 (1987)).
The product was purified using ISOGEN (Nippon Gene Co., Ltd.). In addition, total RN from epidermal keratinocytes and outer root sheath cells
A was prepared similarly. (2) Synthesis of single-stranded cDNA (reverse transcriptase reaction) The synthesis of single-stranded cDNA from purified total RNA can be performed according to a conventional method. Specifically, “2.5 μM Random Hexamers, 5 mM MgCl ₂ , 50 mM KCl,
10 mM Tris-HCl (pH 8.3), 1 mM deoxyribonucleotide triphosphate (dNTP),
1.0 U / μl ribonuclease inhibitor (Ribonuclease Inhib
itor) (Takara Shuzo), 2.5U / μl M-MLV reverse transcriptase (R
everse Transcriptase) (Gibco-BRL)
Was mixed to make a total volume of 100 μl. After denaturation, the tube was set in a thermal cycler, and a synthesis reaction was carried out by the following program. That is, the number of cycles is one, "25 ° C for 15 minutes, 42
At 45 ° C., 5 minutes at 99 ° C., and 10 minutes at 4 ° C. ". (3) PCR (polymerase chain reaction) PCR can be performed according to a conventional method. Specifically, "2
mM MgCl ₂ , 50 mM KCl, 10 mM Tris-HCl (pH 8.3), 0.025 U / μl
Recombinant Taq DNA polymerase
lymerase (Takara Shuzo Co., Ltd.), and a PCR mixture obtained by mixing 5 μl of the sample subjected to the reverse transcriptase reaction into a tube containing 20 μl of a total of 0.5 μM of each primer. As a primer, `` 5'-ACCAACTCTTCTTCCTCCTC-
3 '"(SEQ ID NO: 3) and"5'-CCTCTATCCTCTCCTTCAG
C-3 ′ ”(SEQ ID NO: 4) was used. One drop of mineral oil (Perkin Elmer) was added dropwise to the mixture, and the mixture was set in a thermal cycler.
Cycle, 1 minute at 94 ° C, 1 minute at 60 ° C, 1 minute at 72 ° C
Cycle, "10 minutes at 72 ° C", 1 cycle, "10 minutes at 4 ° C"
Was subjected to one cycle of PCR reaction. After the PCR reaction, the amplification product was visualized by electrophoresis on a 2% agarose gel containing 0.5 μg / ml ethidium bromide.

As a result, expression of FGF10 was observed in hair papilla cells, which are cells of the mesenchymal system (a, b, g, h in FIG. 1A). In addition,
“M” in FIG. 1A indicates a 100 bp molecular weight marker. Also,
"A" and "b" are frontal dermal papilla cells, "c" and "d" are epidermal keratinocytes, "e" and "f" are outer root sheath cells, and "g" and "h" are whisker papilla cells Shows the results of using.

In addition, RT-PCR was similarly performed using hair papilla cells from different subjects (FIG. 1B). In FIG. 1B, “M” indicates a marker, and “a” to “f” indicate the results of experiments on hair papilla cells derived from different subjects. A band was detected at the position of 325 bp in the hair papilla cells of all subjects who performed the experiment. The nucleotide sequence of the amplified fragment is shown in SEQ ID NO: 5.

In order to confirm that the detected band was an FGF-10 band, restriction enzyme digestion analysis was performed. Specifically, a single band confirmed by electrophoresis was cut out, and the DNA was purified using `` The GENECLEAN II Kit '' (BIO 101).
Recovery was performed. The recovered PCR amplification product was subjected to restriction enzyme ScaI.
(Takara Shuzo), and the product was visualized by electrophoresis as described above. As a result, bands of 210 bp and 115 bp which could be expected from the sequence of FGF-10 were observed (FIG. 1).
C). In FIG. 1C, the left lane is a 100 bp molecular weight marker (BRL Life Technology), the middle lane is a sample without ScaI added, and the right lane is a result of electrophoresis of a ScaI digested sample.

Example 3 Preparation of Recombinant FGF10 Protein In order to prepare a recombinant FGF10 protein, first, P
FGF10 cDNA was prepared by CR. Specifically, 5 μl 10
× Pfu buffer (Stratagene), 0.4 mM dNTPmixture
(Takara Shuzo), 5U Pfu DNA polymerase (Stratagene
), 0.5 μM of each primer, 5 μl of the reaction solution after the reverse transcriptase reaction prepared in Example 2 (2), and a total volume of 50 μl of P
A CR mixture was prepared. As the primer, "5'-AAGC
TTATGTGGAAATGGATACTG-3 '"(SEQ ID NO: 6) and"5'-AAGCTTCTATGAGTGTACCACCAT-3'"(SEQ ID NO:
7) was used. The PCR reaction was performed at 94 ° C for 1 minute for 1 cycle.
35 cycles of 72 ° C for 1 minute, 56 ° C for 1 minute, 72 ° C for 2 minutes ”, 72 ° C
For one cycle for 10 minutes and one cycle for 10 minutes at 4 ° C. P
The CR product was subjected to agarose gel electrophoresis, and a 640 bp band was purified and digested with a restriction enzyme HindIII. Human FGF
The HindIII fragment containing the region coding for 10 was inserted into plasmid p.
It was inserted into the HindIII site of Rc / CMV (Invitrogen) to construct pRc / CMV-hFGF10. 24 hours before transfection, a DMEM medium (Gibco B
RL) and COS1 cells (American Type Culture collectio
n) was seeded at 2 × 10 ⁵ cells per 6 cm tissue culture dish. The calcium phosphate method (Graham et a
l, Virolozy, 52, 456, 1973) to increase pRc / CMV-hFGF10 to 5
μg and pRc / CMV without recombinant molecule as control
Was similarly transfected. The next day, the medium was changed to DMEM medium containing 0.1% fetal bovine serum, and the culture was continued for another 40 hours. The transformed COS-1 cells and the culture supernatant were collected. Cells were washed twice with PBS and then washed with 1 M NaCl, 20 mM Tris-HCl (pH =
After suspending in 7.6) and sonication, the mixture was centrifuged at 15000 g at 4 ° C. for 15 minutes to collect only the supernatant. The cell extract and the culture supernatant thus obtained are subjected to ultrafiltration (molecular weight 1).
10,000). The obtained ultrafiltrate was added to solution A (pRc /
CMV-hFGF10 transfected COS1 cells), B
Solution (from COS-1 cells transfected with pRc / CMV)
And

Example 4 Isolation of Human Hair Follicle From a normal scalp obtained at the time of plastic surgery, a conventional method (Phi
lpott, MP et.al., J. Cell.Sci., 97, p463-471 (1
990)). The following operations were all performed under a stereo microscope in a clean bench.

Specifically, the obtained normal scalp was cut into strips having a width of 5 mm or less, and the epidermis and upper dermis were
The hair bulb was separated from the remaining fat layer. Further, under a stereoscopic microscope (Leica WILD MZ8), the adipose tissue containing the hair bulb was placed in a medium, and only the hair follicle was carefully isolated.

Example 5 Method of Organ Follicle Tissue Culture and Evaluation of Hair Follicle Growth As a medium for evaluating hair follicle growth of hair follicle tissue, ultrafiltrate solution A and solution B were each used as a culture medium ( Glutamine (2m
M), insulin (10 μg / ml), transferrin (10 μg / m
l), hydrocortisone (10 ng / ml), selenous acid (10 ng / m
l), penicillin (50 μg / ml), streptomycin (50 IU /
ml) containing William's Medium E (Gibco BRL)
The one that was diluted 10,000 times and 100,000 times was used. Each of the isolated hair follicles obtained in Example 4 was placed in a 12-well culture plate (Corning Coaster Japan) and cultured at 37 ° C., 5% CO ₂ and humidified conditions for 5 days to obtain hair follicle tissue. Follicle growth evaluation was performed. Hair follicle growth was defined as elongation of the entire length of hair follicles cultured for a certain period of time. The evaluation of hair follicle growth was carried out under a microscope using an inverted microscope (Olympus IX70) and a conversion measurement was performed using a measurement scale of an eyepiece. As a result, the hair follicles cultured in the medium in which the ultrafiltrate A solution was diluted 100,000-fold showed an elongation of about 1.3 mm, which was about 15 mm larger than that in the case where the similarly diluted ultrafiltrate B solution was diluted. % Hair follicle growth was promoted. Further, the hair follicles cultured in a medium obtained by diluting the ultrafiltrate A solution 10,000 times have about 1.4
The elongation by mm was observed, and the hair follicle growth was promoted by about 26% similarly to the case where the ultrafiltrate B solution was diluted.

[0030]

Industrial Applicability According to the present invention, it has been revealed that FGF-10 is expressed in hair papilla cells and is a key paracrine factor controlling hair growth, whereby a hair restorer containing FGF-10 as an active ingredient Was provided.

[0031]

[Sequence list]

(1) Applicant's name or name: Rohto Pharmaceutical Co., Ltd. (2) Title of the invention: Hair restorer (3) Reference number: R1-901DP1 (4) Application number: (5) Filing date: (6) Base of priority The name of the application and the application number of the application became: Japanese Patent Application No. 41658, 1997 (7) Priority date: February 10, 1997 (8) Number of sequences: 7 SEQ ID NO: 1 Length of sequence : 627 Sequence type: Number of nucleic acid strands: Double strand Topology: Linear Sequence type: cDNA to mRNA sequence ATG TGG AAA TGG ATA CTG ACA CAT TGT GCC TCA GCC TTT CCC CAC CTG 48 Met Trp Lys Trp Ile Leu Thr His Cys Ala Ser Ala Phe Pro His Leu 1 5 10 15 CCC GGC TGC TGC TGC TGC TGC TTT TTG TTG CTG TTC TTG GTG TCT TCC 96 Pro Gly Cys Cys Cys Cys Cys Cys Phe Leu Leu Leu Phe Leu Val Ser Ser 20 25 30 GTC CCT GTC ACC TGC CAA GCC CTT GGT CAG GAC ATG GTG TCA CCA GAG 144 Val Pro Val Thr Cys Gln Ala Leu Gly Gln Asp Met Val Ser Pro Glu 35 40 45 GCC ACC AAC TCT TCT TCC TCC TCC TTC TCC TCT CCT TCC AGC GCG GGA 192 Ala Thr Asn Ser Ser Ser Ser Ser Phe Ser Ser Pro Ser Ser Ala Gly 50 55 60 AGG CAT GTG CGG AGC TAC AAT CAC CTT CAA GGA GAT GTC CGC TGG AGA 240 Arg His Val Arg Ser Tyr Asn His Leu Gln Gly Asp Val Arg Trp Arg 65 70 75 80 AAG CTA TTC TCT TTC ACC AAG TAC TTT CTC AAG ATT GAG AAG AAC GGG 288 Lys Leu Phe Ser Phe Thr Lys Tyr Phe Leu Lys Ile Glu Lys Asn Gly 85 90 95 AAG GTC AGC GGG ACC AAG AAG GAG AAC TGC CCG TAC AGC ATC CTG GAG 336 Lys Val Ser Gly Thr Lys Lys Glu Asn Cys Pro Tyr Ser Ile Leu Glu 100 105 110 ATA ACA TCA GTA GAA ATC GGA GTT GTT GCC GTC AAA GCC ATT AAC AGC 384 Ile Thr Ser Val Glu Ile Gly Val Val Ala Val Lys Ala Ile Asn Ser 115 120 125 AAC TAT TAC TTA GCC ATG AAC AAG AAG GGG AAA CTC TAT GGC TCA AAA 432 Asn Tyr Tyr Leu Ala Met Asn Lys Lys Gly Lys Leu Tyr Gly Ser Lys 130 135 140 GAA TTT AAC AAT GAC TGT AAG CTG AAG GAG AGG ATA GAG GAA AAT GGA 480 Glu Phe Asn Asn Asp Cys Lys Leu Lys Glu Arg Ile Glu Glu Aslu Gly 145 150 155 160 TAC A AT ACC TAT GCA TCA TTT AAC TGG CAG CAT AAT GGG AGG CAA ATG 528 Tyr Asn Thr Tyr Ala Ser Phe Asn Trp Gln His Asn Gly Arg Gln Met 165 170 175 TAT GTG GCA TTG AAT GGA AAA GGA GCT CCA AGG AGA GGA CAC AAA ACA 576 Tyr Val Ala Leu Asn Gly Lys Gly Ala Pro Arg Arg Gly His Lys Thr 180 185 190 CGA AGG AAA AAC ACC TCT GCT CAC TTT CTT CCA ATG GTG GTA CAC TCA 624 Arg Arg Lys Asn Thr Ser Ala His Phe Leu Pro Met Val Val His Ser 195 200 205 TAG 627 SEQ ID NO: 2 Sequence length: 208 Sequence type: Amino acid Topology: Linear Sequence type: Peptide sequence Met Trp Lys Trp Ile Leu Thr His Cys Ala Ser Ala Phe Pro His Leu 1 5 10 15 Pro Gly Cys Cys Cys Cys Cys Phe Leu Leu Leu Phe Leu Val Ser Ser 20 25 30 Val Pro Val Thr Cys Gln Ala Leu Gly Gln Asp Met Val Ser Pro Glu 35 40 45 Ala Thr Asn Ser Ser Ser Ser Ser Phe Ser Ser Pro Ser Ser Ala Gly 50 55 60 Arg His Val Arg Ser Tyr Asn His Leu Gln Gly Asp Val Arg Trp Arg 65 70 75 80 Lys Leu Phe Ser Phe Thr Lys Tyr Phe Leu Lys Ile Glu Lys Asn Gl y 85 90 95 Lys Val Ser Gly Thr Lys Lys Glu Asn Cys Pro Tyr Ser Ile Leu Glu 100 105 110 Ile Thr Ser Val Glu Ile Gly Val Val Ala Val Lys Ala Ile Asn Ser 115 120 125 Asn Tyr Tyr Leu Ala Met Asn Lys Lys Gly Lys Leu Tyr Gly Ser Lys 130 135 140 Glu Phe Asn Asn Asp Cys Lys Leu Lys Glu Arg Ile Glu Glu Asn Gly 145 150 155 160 Tyr Asn Thr Tyr Ala Ser Phe Asn Trp Gln His Asn Gly Arg Gln Met 165 170 175 Tyr Val Ala Leu Asn Gly Lys Gly Ala Pro Arg Arg Gly His Lys Thr 180 185 190 Arg Arg Lys Asn Thr Ser Ala His Phe Leu Pro Met Val Val His Ser 195 200 205 SEQ ID NO: 3 Sequence length: 20 Sequence length Type: Number of nucleic acid strands: Single strand Topology: Linear Sequence type: Other nucleic acid Synthetic DNA sequence ACCAACTCTT CTTCCTCCTC 20 SEQ ID NO: 4 Sequence length: 20 Sequence type: Number of nucleic acid strands: Single Chain topology: linear Sequence type: Other nucleic acid Synthetic DNA sequence CCTCTATCCT CTCCTTCAGC 20 SEQ ID NO: 5 Sequence length: 325 Sequence type: Nucleic acid Number of strands: Double-stranded Topology: linear sequence type: cDNA-to mRNA sequences ACCAACTCTT CTTCCTCCTC CTTCTCCTCT CCTTCCAGCG CGGGAAGGCA TGTGCGGAGC 60 TACAATCACC TTCAAGGAGA TGTCCGCTGG AGAAAGCTAT TCTCTTTCAC CAAGTACTTT 120 CTCAAGATTG AGAAGAACGG GAAGGTCAGC GGGACCAAGA AGGAGAACTG CCCGTACAGC 180 ATCCTGGAGA TAACATCAGT AGAAATCGGA GTTGTTGCCG TCAAAGCCAT TAACAGCAAC 240 TATTACTTAG CCATGAACAA GAAGGGGAAA CTCTATGGCT CAAAAGAATT TAACAATGAC 300 TGTAAGCTGA AGGAGAGGAT AGAGG 325 SEQ ID NO: 6 Sequence length: 24 Sequence type: Number of nucleic acid strands: Single strand Topology: Linear Sequence type: Other nucleic acid Synthetic DNA sequence AAGCTTATGT GGAAATGGAT ACTG 24 SEQ ID NO: 7 Sequence Length: 24 Sequence type: Nucleic acid number of strands: Single-stranded Topology: Linear Sequence type: Other nucleic acid Synthetic DNA sequence AAGCTTCTAT GAGTGTACCA CCAT 24

[Brief description of the drawings]

FIG. 1A shows RT-PCR of FGF10 expression in frontal hair papilla cells, epidermal keratinocytes, outer root sheath cells, and hair papilla cells.
5 is an electrophoresis image detected by the method shown in FIG. B is an electrophoresis image in which expression of FGF10 in hair papilla cells from different subjects was detected by RT-PCR. C is FG treated with restriction enzyme
It is an electrophoresis image of F-10.

Claims (1)

[Claims] 1. A hair restorer comprising FGF-10 as an active ingredient.