JP5017535B2 - Composition for inhibiting hair loss or promoting hair growth - Google Patents

Description

  The present invention relates to a composition for effectively preventing or treating hair loss.

  Hair is a part of the skin that changes with the development of the fetus along with the sebaceous glands and sweat glands. Accordingly, the hair is generated with the development of the skin, and the keratinization progresses to become one thick hair. Hair grows when hair matrix cells present in the hair papilla in the hair root part divide and proliferate while synthesizing keratin protein and push the hair shaft upward. Capillaries are stretched around the bottom of the hair papilla, and supplementation of oxygen and nutrients from the blood vessels supports the growth of hair matrix cells. Hair has a certain life span, and after it grows up, it naturally comes off and new hair grows again. This is called a hair cycle, and this hair cycle is divided into three phases: a growth phase, a regression phase, and a resting phase.

  However, if the balance of the hair cycle is lost due to various causes such as poor circulation to the hair papilla and hair follicles, activation of male hormones in the hair follicles and sebaceous glands, excessive secretion of sebum, nutritional disorders and stress, In this process, abnormal hair loss occurs in which hair that should not be removed comes out earlier than the lifetime.

  The mechanism of hair loss has not yet been fully elucidated, but hair loss due to excessive secretion of sebum is considered as follows. A square plug is formed at the entrance of the pore due to the action of the skin resident bacteria and the oxidation of sebum secreted from the pore, thereby closing the pore. The sebum in the pores is not excreted outside the skin, but becomes a fat plug in the pores, reducing the root fixing force. For this reason, not only existing hair but also new hair is easily removed even by a stimulus such as washing or brushing. Thus, although the life of new hair is originally 3 to 6 years in the case of humans, the hair growth cycle (hair cycle) is shortened, and abnormalities that come out before the hair grows sufficiently Hair loss occurs. Furthermore, excessively secreted sebum also slows down cell division of hair matrix cells in the pores and inhibits hair formation. In modern times, such an excessive secretion of sebum is likely to occur due to an irregular life and an unbalanced diet.

  Pigment epithelium-derived factor (PEDF) is a glycoprotein belonging to the superfamily of serine protease inhibitors and was first purified from a conditioned medium of human retinal pigment epithelial cells as a factor having potent human retinoblastoma neuronal differentiation activity. (Non-Patent Document 1).

  PEDF has been suggested to be a potent inhibitor of angiogenesis in both cell culture and animal models. PEDF has been reported to inhibit proliferation and migration of retinal endothelial cells and suppress ischemia-induced retinal neovascularization (Non-patent Documents 2 and 3). Furthermore, the loss of PEDF is associated with angiogenic activity in proliferative diabetic retinopathy (Non-Patent Document 4). PEDF is also known to effectively suppress retinal and choroidal neovascularization due to ischemia and age-related macular degeneration (Non-patent Documents 3 and 5). It has been found that PEDF can be used for the treatment and prevention of vascular disorders such as diabetes and atherosclerosis by suppressing NADPH oxidase activity (Patent Document 1).

Recently, it has also been reported that PEDF-deficient mice are susceptible to pancreatic cancer (Non-Patent Document 6). PEDF has been disclosed to up-regulate Fas ligand in endothelial cells, thereby making tumor blood vessels specifically apoptotic (Non-Patent Document 7). Thus, PEDF suppresses tumor angiogenesis, induces apoptosis in cancer cells, and suppresses the growth of cancer cells such as malignant melanoma, and thus is seen to be effective in the prevention or treatment of cancer. (Patent Document 2).
JP 2005-336159 A JP 2005-298473 A US Pat. No. 6,319,687 International Publication No. 03/059248 Pamphlet International Publication No. 93/24529 Pamphlet Tombran-Tink, J. et al., Exp. Eye Res., 53, 411-414 (1991) Dawson, DW et al., Science, 285, 245-248 (1999) Duh, EJ et al., Invest. Ophthalmol. Vis. Sci., 43, 821-829 (2002) Spranger, J. et al., Diabetes, 50, 2641-2645 (2001) Holekamp, NM et al., Am. J. Ophthalmol., 134, 220-227 (2002) Doll, JA et al., Nat. Med., 9, 774-780 (2003) Volpert, OV et al., Nat. Med., 8, 349-357 (2002) Steel, FR et al., Proc. Natl. Acad. Sci. USA, 90 (4), 1526-1530 (1993) Yamagishi, S. et al., Biochem. Biophys. Res. Commun., 296 (4), 877-882 (2002)

  An object of the present invention is to provide a composition for effectively preventing or treating hair loss.

The present invention provides a composition for inhibiting hair loss. This hair loss inhibiting composition is
(A) pigment epithelium-derived factor,
(B) a variant of the factor having functionally equivalent properties to the pigment epithelium-derived factor (a), and
(C) containing an active ingredient selected from the group consisting of a vector comprising a nucleic acid molecule encoding the factor (a) or the mutant (b).

The present invention also provides a composition for promoting hair growth. This hair growth promoting composition is
(A) pigment epithelium-derived factor,
(B) a variant of the factor having functionally equivalent properties to the pigment epithelium-derived factor (a), and
(C) containing an active ingredient selected from the group consisting of a vector comprising a nucleic acid molecule encoding the factor (a) or the mutant (b).

The present invention further provides a vector for suppressing hair loss. This vector
A nucleic acid molecule encoding a variant of the factor having (a) a pigment epithelium-derived factor, or (b) a functionally equivalent property of the pigment epithelium-derived factor (a).

The present invention further provides a vector for promoting hair growth. This vector
A nucleic acid molecule encoding a variant of the factor having (a) a pigment epithelium-derived factor, or (b) a functionally equivalent property of the pigment epithelium-derived factor (a).

  According to the present invention, a composition for effectively preventing or treating hair loss is provided. The composition of the present invention is useful for preventing and treating hair loss even when sebum is secreted excessively. Furthermore, it is useful for promoting hair growth.

  The present invention provides a composition for suppressing hair loss and a composition for promoting hair growth. “A composition for suppressing hair loss” refers to a composition having an action of suppressing hair loss, and this composition suppresses the occurrence of hair loss when, for example, excessive secretion of sebum occurs. The “hair growth promoting composition” refers to a composition having an effect of causing hair growth, and this composition can promote hair formation and growth.

(1) Active ingredient contained in the composition of the present invention In the composition of the present invention, the active ingredient is selected from the group consisting of:
(A) pigment epithelium-derived factor,
(B) a variant of the factor having functionally equivalent properties to the pigment epithelium-derived factor (a), and
(C) A vector comprising a nucleic acid molecule encoding the factor (a) or the mutant (b).

(1-1) Protein In the present invention, the term “pigment epithelium-derived factor” or “PEDF” simply means pigment epithelium-derived factor protein. PEDF has hair loss inhibiting activity and / or hair growth promoting activity. Furthermore, PEDF can prevent apoptosis of cells in the hair root including hair matrix cells.

  In the present invention, PEDF includes all kinds of PEDFs derived from mammals such as humans, dogs, cats, cows, and horses, and human-derived PEDF is preferable. The amino acid sequence of human PEDF and the nucleic acid sequence encoding the same are described in Non-Patent Document 8, and registered in the GenBank / EMBL database under accession no. M76979.

  PEDF encodes the protein according to procedures described in many publications and references such as Molecular Cloning: A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, (1989) It can be produced by expressing DNA. More specifically, an expression plasmid is constructed by inserting DNA encoding the protein into an appropriate expression vector (for example, pBK-CMV). Then, the expression plasmid is introduced into an appropriate host cell to obtain a transformed cell. Examples of host cells include prokaryotes such as E. coli, unicellular eukaryotes such as yeast, and multicellular eukaryotes such as insects and animals. Expression plasmids can be introduced into host cells by methods commonly used by those skilled in the art, such as the calcium phosphate method, electroporation method, lipofectin method and the like. The desired protein is produced by culturing the transformant in an appropriate medium according to a method commonly used by those skilled in the art. The protein thus obtained can be isolated and purified according to standard biochemical techniques.

  As used herein, “variants of PEDF having functionally equivalent properties to PEDF” include all types of PEDF variants having functionally equivalent properties to human PEDF. PEDF variants are described in, for example, Patent Documents 3 to 5.

  Preferred examples of PEDF variants include substitution, deletion, and / or addition of one or more amino acid residues to the amino acid sequence of human PEDF (eg, the amino acid sequence described in Non-Patent Document 8 above). And a variant of PEDF having a functionally equivalent property to human PEDF. A person skilled in the art can modify the structure of a protein by appropriately introducing substitution, deletion, and / or addition mutations using techniques usually used by those skilled in the art such as site-directed mutagenesis. Can do. In the present invention, the number of amino acid residues that can be substituted, deleted, and / or added is usually 120 or less, such as 100 or less, 80 or less, 50 or less, or 20 or less, preferably 16 or less, more preferably 5 Hereinafter, 0 to 3 amino acid residues are more preferable. In addition, since amino acid mutations may occur in nature, the present invention is not limited to proteins in which amino acids are artificially mutated, and proteins in which amino acids are mutated in nature have the same functional characteristics as human PEDF. Can be used in the composition.

  As long as the protein having an amino acid sequence homologous to the amino acid sequence of human PEDF (for example, the amino acid sequence described in Non-Patent Document 8) also has functionally equivalent characteristics to human PEDF, the composition of the present invention Can be used for The PEDF variant preferably has at least 70%, preferably at least 80%, more preferably at least 90%, even more preferably at least 95%, even more preferably at least 99% homology with the human PEDF amino acid sequence. It may be a protein having an amino acid sequence having Protein homology searches include, for example, databases related to amino acid sequences of proteins such as SWISS-PROT, PIR, and DAD, databases related to DNA sequences such as DDBJ, EMBL, and GenBank, and deduced amino acid sequences based on DNA sequences. For example, it can be performed over the Internet using a data analysis program such as BLAST or FASTA for a database or the like. Such PEDF variants can also be prepared by the recombinant techniques described above.

  Properties functionally equivalent to human PEDF have hair loss inhibiting activity or hair growth promoting activity equivalent to that of human PEDF, or can prevent apoptosis of hair root cells including hair matrix cells as well as human PEDF. Means that.

  Whether the mutant prepared as described above has functionally equivalent properties as PEDF can be proved according to the examples described later in this specification.

(1-2) Nucleic Acid Molecules and Vectors As used herein, the term “nucleic acid molecule” includes DNA and RNA that can be single-stranded or double-stranded. Nucleic acid molecules are described in standard textbooks such as typical DNA synthesis or genetic engineering methods such as Molecular Cloning: A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, (1989). It is easily prepared by the method.

  As described above, the sequence of the nucleic acid molecule encoding human PEDF is described in Non-Patent Document 8, and registered with GenBank / EMBL accession no. M76979. Preferably, a nucleic acid molecule having the nucleic acid sequence described in Non-Patent Document 8 is used in the composition of the present invention.

  Examples of the nucleic acid molecule used in the composition of the present invention also include a nucleic acid molecule that encodes a protein having functionally equivalent characteristics to the human PEDF of (1-1) above. A person skilled in the art can appropriately introduce substitution, deletion, and / or addition mutations into the nucleic acid sequence described in Non-Patent Document 8 using techniques usually used by those skilled in the art, such as site-directed mutagenesis. Polynucleotide homologues can be obtained.

  Such a nucleic acid molecule can be administered to a subject such that PEDF or a variant thereof is expressed in the subject and exhibits hair loss inhibiting activity or hair growth promoting activity. For this purpose, the nucleic acid molecule can be incorporated into a vector and administered in vivo.

  As a vector for incorporating a nucleic acid molecule, a conventionally useful vector for gene therapy can be used. Such vectors include, for example, retroviral, adenovirus, adeno-associated virus, herpes virus, vaccinia virus, poxvirus, poliovirus or Sindbis virus or a nucleic acid molecule incorporated into a cell after introduction into a cell. A virus vector. Of these, those using retroviruses, adenoviruses, adeno-associated viruses or herpes viruses are particularly preferred. Specific examples of the vector include pAd / CMV / V5-DEST Gateway vector (Invitrogen).

  The present invention also includes vectors themselves incorporating such nucleic acid molecules.

  For example, the vector of the present invention can be generated by the following procedure. PEDF cDNA can be isolated from a human placenta cDNA library (available from, for example, Clontech, Palo Alto, Calif.), For example, as described in [9]. This can be incorporated into a commercially available adenoviral vector (eg, pAd / CMV / V5-DEST Gateway vector (Invitrogen)) using methods commonly used by those skilled in the art. Commercially available vectors incorporating the PEDF gene can also be used for administration of PEDF or variants thereof.

(2) Composition of the present invention The composition of the present invention contains the active ingredient of the above (1) and exhibits an action of suppressing hair loss. The composition of the present invention also exhibits an effect of promoting hair growth. Furthermore, apoptosis or cell death of cells in the hair root including hair matrix cells can be prevented.

  In addition to preventing hair loss and promoting hair growth in human scalp, the composition of the present invention is also used for preventing hair loss and promoting hair growth in animals (for example, hair growth in fur animals such as mink, hair loss prevention in pet animals, etc.) It can be used. The composition of the present invention may exhibit a hair growth action even under conditions where the excessive secretion of sebum that has caused hair loss continues. Therefore, it is particularly useful for the prevention and treatment of hair loss that occurs with excessive secretion of sebum.

  As long as the composition of this invention can exhibit the effect, a dosage form and an administration route are not specifically limited. Usually, it is mixed with physiologically acceptable carriers, excipients and the like to be in a form suitable for oral or parenteral administration or application.

  When the active ingredient in the composition of the present invention is a protein, it may be oral or parenteral. For example, it may be administered by means such as direct application, spraying, spraying, or subcutaneous injection to a site where hair loss is suppressed or hair growth is desired.

  When the active ingredient in the composition of the present invention is a vector, it can be administered to a subject by injection (intravenous injection, intramuscular injection, subcutaneous injection, etc.). In the case of humans, it is desirable that the nucleic acid molecule incorporated in the vector is expressed as a protein that suppresses hair loss in the scalp or promotes hair growth in the hair loss part of the scalp.

  Specific examples of dosage forms for oral administration include tablets, pills, capsules, granules, syrups, emulsions and suspensions. Such dosage forms are produced by methods commonly used by those skilled in the art and may contain carriers or excipients commonly used in the pharmaceutical field.

  Examples of the dosage form for parenteral administration or application include ointments, injections, poultices, coating agents, transdermal absorption agents, topical sustained release agents, and the like. The above dosage forms are produced by methods commonly used by those skilled in the art and may contain carriers or excipients commonly used in the pharmaceutical field. The composition of the present invention can also be processed into various forms such as lotions, emulsions, gels, creams, ointments and the like.

  Solution preparations such as injections are usually used by those skilled in the art, for example, in a state where the active ingredient is dissolved or suspended in a sterile aqueous solution usually used for injections, or further emulsified and encapsulated in liposomes. Can be prepared. The solid preparation can be prepared as a lyophilized product by adding a mannitol, trehalose, sorbitol, lactose, glucose, starch or the like to the active ingredient as an excipient by a method usually used by those skilled in the art. Further, it can be used in the form of powder. Further, these powders may be mixed with polylactic acid or glycolic acid to be solidified. The gelling agent can be prepared by a method commonly used by those skilled in the art, for example, in a state where the active ingredient is dissolved in a thickener or polysaccharide such as glycerin, polyethylene glycol, methylcellulose, carboxymethylcellulose, hyaluronic acid, chondroitin sulfate, and the like.

  In any of the above preparations, alcohols, sugar alcohols, ionic surfactants, nonionic surfactants, and the like can be added as dispersants or absorption promoters within a range that does not impair the physiological activity of the active ingredient. Trace metals and organic acid salts can also be added as necessary.

  As long as the effect of the active ingredient in the composition of the present invention is not diminished, other components commonly used by those skilled in the art, for example, antioxidants, preservatives, coloring agents, stabilizers, solubilizing agents, You may add a viscosity modifier, a refreshing agent, a fragrance | flavor, etc.

The dose can be appropriately determined according to the age, weight, sex, administration target disease, symptom, administration mode, administration route, etc. of the administration subject. When the active ingredient is protein, typically 0.0001 mg to 1000 mg, preferably 0.001 mg to 100 mg, more preferably 0.01 mg to 10 mg, and when the active ingredient is a vector, typically 1 × 10 ^6. ˜1 × 10 ⁹ pfu, preferably 5 × 10 ^{6 to} 5 × 10 ⁸ pfu, more preferably 1 × 10 ⁷ to 1 × 10 ⁸ pfu of active ingredient daily (from once to several times a day) Or can be administered or applied every few days to every few months. Therefore, the active ingredient in the composition of the present invention is contained in the preparation so that it can be administered or applied in such an amount.

  The composition of the present invention can be used in combination with other known hair growth promoters, hair growth agents, hair nourishing agents, and the like.

  The present invention will be described in more detail with reference to the following examples, which are not intended to limit the present invention in any way.

In the examples, the commercially available vector AdenoVec ^™ -hPEDF (purchased from InvivoGen) was used to administer PEDF to mice. This is a vector having an adenovirus genome as a backbone and a human PEDF gene. Hereinafter, this vector is referred to as a PEDF expression adenovector. As a control, phosphate buffered saline (PBS) or adenovirus vector pAd-LacZ (purchased from Invitrogen) was administered. Administration was by intravenous injection from the tail vein or intramuscular injection into the thigh muscle.

(Example 1: Hair loss inhibitory activity of PEDF)
C57BL6 / J mice (purchased from CLEA) having a body weight of 18 to 20 g and aged 6 to 8 weeks were bred for 10 weeks under conditions free of specific pathogens. The breeding method was carried out according to a protocol approved in accordance with the guidelines of Kurume University Animal Experiment Ethics Committee or the United States Department of Agriculture (USDA) “the Guide for the Care and Use of Laboratory Animals”.

The mice were randomly divided into 12 groups of about 20-30 animals. These groups were as follows:
1. PEDF-expressing adenovector was injected intravenously and fed a choline-deficient diet;
2. PEDF-expressing adenovector was injected intramuscularly and fed a choline-deficient diet;
3. pAd-LacZ was injected intravenously and fed a choline-deficient diet;
4). injected intramuscularly with pAd-LacZ and fed a choline-deficient diet;
5. PBS was injected intravenously and fed a choline-deficient diet;
6). PBS was injected intramuscularly and fed a choline-deficient diet;
7). PEDF-expressing adenovector was injected intravenously and not fed a choline-deficient diet;
8). PEDF-expressing adenovector was injected intramuscularly and not fed a choline-deficient diet;
9. pAd-LacZ was injected intravenously and was not fed a choline-deficient diet;
10. pAd-LacZ was injected intramuscularly and not fed a choline-deficient diet;
11. 11. PBS injected intravenously and not fed choline deficient diet; PBS was injected intramuscularly and was not fed a choline-deficient diet.

In the choline-deficient diet-fed group, mice were allowed to eat a low protein, high fat choline-deficient diet from the start of the study. The composition of this choline-deficient diet (unit:% by weight) is: vitamin-free casein 8.000; lard 37.950; granulated sugar 48.375; L-cystine 0.625; vitamin Mix 1.000; mineral Mix 4.000; Oily vitamin mixture 0.050. Here, the vitamin Mix has the following composition (unit: mg / feed g): vitamin K3 0.3125, vitamin B1 0.3125, vitamin B2 0.5000, vitamin B6 0.3125, DL-pantothenic acid Ca 2500, niacin 1.2500, the remainder has granulated sugar, and the mineral Mix has the following composition (unit: mg / 100 g of feed): Ca (H ₂ PO ₄ ) H ₂ O 585.000, lactate Ca 1, 328.000, Fe citrate 118.800, MgSO ₄ · 7H ₂ O 548.000, K ₂ HPO ₄ 959.200, NaH ₂ PO ₄ · 2H ₂ O 453.600, and NaCl 174,000. The oily vitamin mixture has the following composition (unit: mg / 100 g of feed): Vitamin A (500,000 IU / g) 2.50 0, vitamin D3 (50 million in IU / g) 0.6250, having vitamin E 25.0000, and peanut oil 21.8750. In the group not fed the choline-deficient diet, a normal diet was given.

On the test start day (day 0), a PEDF expression adenovector, an adenovirus vector pAd-LacZ, or PBS was administered. Regardless of administration route, PBS is 100 μl, adenoviral vector pAd-LacZ 1 × 10 ⁸ pfu (with PBS to 100 μl), PEDF expression adenovector 1 × 10 ⁸ pfu (with PBS to 100 μl) The dose was administered. From the day after the administration, the condition of the mouse hair was observed once a day. Five weeks after the start of the test (gene expression by the vector decreases around this time), mice that continue to receive a choline-deficient diet are again treated with PEDF-expressing adenovector or PBS or adenoviral vector pAd-LacZ as a control. Administered. Observation was continued until 10 weeks after the start of the test. Hereinafter, a mouse administered with a PEDF-expressing adenovector may be referred to as a PEDF-administered mouse, and a mouse administered with PBS or pAd-LacZ may be referred to as a control mouse.

  FIG. 1 is a photograph showing body hair states of PEDF-administered mice (PEDF group; 4 mice on the left side) and control mice (Control group; 4 mice on the right side) 6 weeks after the start of feeding choline-deficient diet. is there. The mouse normally has bushy black body hair. In the group that did not receive the choline-deficient diet, all had black body hair. However, as hair loss occurs, the body color becomes lighter and the skin color becomes exposed. The PEDF group had soft black body hair, while the control group to which PBS or pAd-LacZ was administered had lighter back hair and exposed skin. In the Control group, hair loss was observed 2 weeks after the start of feeding choline-deficient diet. Thus, the suppression of hair loss was clearly observed in the PEDF group. The results were similar for both intravenous and intramuscular administration.

  Further, the skin of the hair loss part of the control mouse and the corresponding part of the PEDF-administered mouse (hair loss did not occur) was peeled off, a paraffin-embedded section thereof was prepared, and histological observation was performed. Skin section specimens were stained with hematoxylin and eosin. In this staining, the cell nucleus is stained blue-purple, and the other cytoplasm is stained red. Staining was carried out by the following procedure: (1) Washing with deparaffin and distilled water, (2) Soaking in hematoxylin solution of caratchi for 20 minutes, (3) Washing lightly, (4) 70% alcohol solution containing 1% hydrochloric acid (5) Wash with water for 10 minutes after color development, (6) Lightly pass through 80% ethanol, then soak in eosin solution for 10 minutes, (7) Lightly wash with water, (8) Dehydrate, clear, and enclose.

  FIG. 2 shows micrographs of the hair loss part of a control mouse stained with hematoxylin and eosin (HE) and the skin section of a PEDF-treated mouse. In a skin section of a PEDF-administered mouse (PEDF group), it was found that the hair roots were stained lightly black and the hair matrix cells were alive. On the other hand, in the skin section of the control mouse (Control group), only the remnants of the hair roots were seen, suggesting that the hair matrix cells were dead for some reason.

  Separately, skin section specimens were also stained with Apop-Tag and methyl green. With Apop-Tag, the nuclei of apoptotic cells are stained brown. In methyl green, DNA in the nucleus is dyed greenish blue and RNA in the nucleolus is dyed red, so that the cell nucleus is dyed brown. Apop-Tag staining was performed using an ApopTag (registered trademark) In Situ Apoptosis detection kit (available from Chemicon). In this kit, the DNA fragment ends of apoptotic cells are labeled in situ using TdT (terminal deoxynucleotidyl transferase), and apoptotic cells are detected by specific staining. Methyl green staining was performed by the following procedure: (1) Deparaffinization and washing with distilled water, (2) 10-15 minutes soaked in methyl green pyronin solution, (3) Hurry with distilled water and rinse with slide (4) Separation with n-butanol I / II / III, dehydration, (5) clearing and sealing.

  In the control mouse (Control group), it was found that there was a lot of brown over the epidermal layer, and the cells were apoptotic. From the result of nuclear staining, it was found that cells including hair matrix cells were broken. On the other hand, apoptosis was hardly observed in PEDF-administered mice (PEDF group). Nuclear staining results showed that the cells were ordered (data not shown).

(Example 2: Hair growth promoting activity of PEDF)
Since it was found that PEDF suppresses hair loss in Example 1 above, in this example, it was examined whether PEDF could promote new hair growth from a state where hair was once removed.

In the same manner as in Example 1, the mice were fed a choline-deficient diet for 14 days to cause hair loss. Four weeks after the start of feeding choline-deficient diet, PEDF-expressing adenovector 1 × 10 ⁸ pfu (with a volume of 100 μl with PBS) or 100 μl of PBS or adenoviral vector pAd-LacZ 1 × 10 subcutaneously in the hair loss site. ⁸ pfu (with 100 μl volume with PBS) was administered once.

  FIG. 3 is a photograph showing the state of body hair before administration of a PEDF-expressing adenovector (left) and 2 weeks after administration of a PEDF-expressing adenovector (right) in mice continuously taking a choline-deficient diet. In the mouse before administration of the PEDF-expressing adenovector, hair loss occurred, and thus the color of the hair became lighter and the part where the skin was exposed was seen. On the other hand, in the mice after administration of the PEDF-expressing adenovector, hair was growing, and a portion that recovered until black body hair grew was also observed. As described above, it was confirmed that, even though the choline-deficient diet was continuously ingested, by the administration of the PEDF-expressing adenovector, hair was gradually grown around the administered portion. On the other hand, no hair growth was observed in control mice (Control group: PBS or adenovirus vector Ad-LacZ administered) (data not shown).

  According to the present invention, a composition for effectively preventing or treating hair loss is provided. The composition of the present invention can be used for preventing hair loss and promoting hair growth in human scalp and preventing hair loss and promoting hair growth in animals (for example, hair growth in fur animals such as mink, hair loss prevention in pet animals, etc.) It is. It is particularly useful for the prevention and treatment of hair loss that occurs with excessive secretion of sebum.

It is a photograph which shows the state of the body hair of the PEDF administration mouse | mouth (PEDF group) and the control mouse | mouth (Control group) 6 weeks after a choline deficient diet start. It is a microscope picture of the skin section dye | stained with hematoxylin and eosin (HE) in a PEDF administration mouse | mouth (PEDF group) and a control mouse | mouth (Control group). It is a photograph which shows the state of the body hair before administration of a PEDF expression adenovector and 2 weeks after PEDF expression adenovector administration in the mouse | mouth which is continuously ingesting a choline deficient diet.

Claims (4)

A composition for inhibiting hair loss caused by excessive secretion of sebum,
(A) pigment epithelium-derived factor,
(B) an amino acid sequence having at least 90% identity with human pigment epithelium-derived factor, or substitution of 50 or fewer amino acid residues to the amino acid sequence of the human pigment epithelium-derived factor, deletions, and / or addition has an amino acid sequence having and having an equivalent hair loss inhibiting activity and dye epithelium-derived factor (a), variant of the dye epithelium-derived factor (a), and <br/> (c) the factor A composition comprising an active ingredient selected from the group consisting of (a) or a vector comprising a nucleic acid molecule encoding the variant (b).   The composition according to claim 1, wherein the suppression of hair loss is suppression of hair loss by preventing apoptosis of hair matrix cells.   The composition according to claim 1 or 2, which is for subcutaneous administration to a hair loss site.   The composition according to any one of claims 1 to 3, wherein the active ingredient is the vector of (c), and the vector is an adenovirus vector.

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