WO2008026349A1

WO2008026349A1 - Therapeutic agent for telogen effluvium

Info

Publication number: WO2008026349A1
Application number: PCT/JP2007/059096
Authority: WO
Inventors: Satoshi Itami; Yasuyuki Sumikawa
Original assignee: Juridical Foundation Osaka Industrial Promotion Organization; Osaka University
Priority date: 2006-08-31
Filing date: 2007-04-26
Publication date: 2008-03-06
Also published as: JPWO2008026349A1; JP5051471B2

Abstract

A therapeutic agent for telogen effluvium, comprising leptin or a partial peptide thereof as an active ingredient. The therapeutic agent is useful for changing the telogen phase in the hair growth cycle to the anagen phase to treat telogen effluvium (e.g., male pattern baldness).

Description

Specification

Resting phase alopecia treatment

Technical field

[0001] The present invention relates to a therapeutic agent for resting alopecia, and more specifically to a therapeutic agent for resting alopecia containing lebutin or a partial peptide thereof as an active ingredient.

Background art

[0002] Hair is distinguished from head hair, eyebrows, eyelashes, beards, eyelashes, pubic hairs, body hairs, and the like, depending on the site. However, the structure is common, and a single hair grows in the hair matrix of the root of the root by the cell division, and a new hair component is born and grows, and is supported by the inherent epithelial hair follicle that envelops it. Fixed.

[0003] Hair growth in the scalp is not caused continuously, but is caused by an active cycle in which the period of growth and rest is alternately repeated. This cycle is mainly divided into three stages: the growth period, the regression period, and the rest period, and this is called the hair cycle. Growth is the developmental stage of the cycle, characterized by deep penetration of hair follicles into the dermis, rapid proliferation and differentiation of cells to form hair. The next stage is the regression phase, which is a transitional stage characterized by the termination of cell division, during which the hair follicles regress in the dermis and the hair growth stops. The next quiescent phase is characterized as a quiescent phase, during which the regressed hair follicle has a hair root in close proximity to the dermal papillary cell population. Thereafter, cells rapidly grow in the hair root, the dermal papilla expands, the basement membrane components become assimilated, and a new growth phase begins. By the way, if sufficient growth is not maintained and the diapause is shifted to the resting phase, so-called resting alopecia occurs.

[0004] Currently, as a therapeutic agent for such resting alopecia, topical minoxidil (ROGAINE (registered trademark) from Pharmacia & Upjohn) for the treatment of male alopecia, a kind of resting alopecia And finasteride (or Merck & Co., Inc. is also commercially available as PROPECI A®). In Patent Document 1, administration of nicotinic acid ester increases the level of levtin, which also causes skin tumor degeneration, skin homoostasis and reconstruction, wound healing, and hair growth. It describes what to do. However, this document does not describe that levatin can be treated by inducing the growth phase from the resting phase of the hair cycle by administering levtin.

[0005] Patent Document 1: Special Table 2004-519488

Disclosure of the invention

Problems to be solved by the invention

[0006] An object of the present invention is to provide a drug useful for treating resting alopecia (especially male pattern alopecia) by inducing the growth phase from the resting phase of the hair cycle.

Means for solving the problem

[0007] As a result of intensive studies to solve the above problems, the present inventors have found that lebutin or a partial peptide thereof induces the growth phase from the resting phase of the hair cycle, thereby treating resting alopecia. We have found that this product has an excellent effect, and based on this knowledge, we have further researched and completed this invention.

That is, the present invention provides:

[I] A therapeutic agent for resting alopecia characterized by containing levtin or a partial peptide thereof as an active ingredient,

[2] The therapeutic agent according to the above [1], wherein the active ingredient is levutin,

[3] The therapeutic agent according to the above [1], wherein the active ingredient is a partial peptide of lebutin,

[4] The therapeutic agent according to the above [2], wherein the lebutin is human leptin,

[5] The therapeutic agent according to [4] above, wherein the human leptin is human recombinant levbutin,

[6] The therapeutic agent according to the above [3], wherein the partial peptide is represented by SEQ ID NO: 4,

[7] The therapeutic agent according to any one of [1] to [6] above, wherein the resting alopecia is androgenetic alopecia,

[8] The therapeutic agent according to any one of the above [1] to [7], wherein the formulation type is an injection for subcutaneous administration

[9] The therapeutic agent according to any one of the above [1] to [7], wherein the formulation type is a preparation for topical application, [10] The therapeutic agent according to the above [9], wherein the active ingredient is ribosomeized,

[II] Lebutin or a partial peptide thereof for producing a therapeutic agent for resting alopecia Use, and

[12] A method for treating resting alopecia characterized by administering a therapeutically effective amount of levbutin or a partial peptide thereof to a patient with resting alopecia,

About.

The invention's effect

[0009] The therapeutic agent for alopecia alopecia according to the present invention can be administered to humans to induce the hair cycle from the resting phase to the growing phase to treat resting alopecia.

Brief Description of Drawings

[0010] FIG. 1 is a photograph showing a hair cycle (A) and a hair follicle state of a 35-day-old (growth stage) of dbZdb mice or normal mice. B represents a normal mouse and C represents a dbZdb mouse.

FIG. 2 shows photographs of the 14th day after back hair removal of obZob mice or normal mice at 7 weeks of age (second resting stage) and the hair follicle state of obZob mice or normal mice. A and B are normal mice, and C and D are obZob mice.

Figure 3 is a photograph of the dorsal part of the 16th day of ob / ob mice administered with saline or leptin. A shows a physiological saline-administered mouse, and B shows a levulin-administered mouse.

Figure 4 shows photographs of the hair follicle state on day 21 after administration of normal saline or levutin to 7-week-old normal mice, and photographs of the back of day 21 of normal mice administered with levutin or physiological saline. It is. A shows a saline-administered mouse, and B shows a lebutin-treated mouse. Above C shows leptin-administered mice and below C shows saline-administered mice.

FIG. 5 is a photograph of the dorsal region on the 21st day of a normal mouse in which physiological saline or a partial peptide of lebutin (SEQ ID NO: 4) was administered to a normal mouse 7 weeks old.

BEST MODE FOR CARRYING OUT THE INVENTION

[0011] The active ingredient of the therapeutic agent for alopecia according to the present invention is lebutin or a partial peptide thereof. The leptin according to the present invention may be mouse leptin or human leptin, preferably human leptin. The amino acid sequence of human leptin including the signal peptide (21 amino acids) is shown in SEQ ID NO: 1, and that of mouse is shown in SEQ ID NO: 2.

[0012] In addition, the levtin according to the present invention may be substituted by substitution or addition of several amino acids as long as the levtin function is maintained. [0013] On the other hand, the partial peptide of levulin according to the present invention includes a partial peptide containing the levtin active site, that is, a low molecular functional peptide. Specific examples of partial peptides that are preferred in that they are low in antigenicity due to partial peptide strength generated by cleaving with an enzyme originally possessed by an animal (particularly human) that is the origin of levutin include SEQ ID NO: 4 The peptide shown by these is mentioned.

[0014] The lebutin and the partial peptide of lebutin according to the present invention can be prepared by a conventional method, and commercially available products such as human recombinant leptin (R & D Systems) or partial peptide of human leptin (BRACHEM) should be used. You can also. In the case of production, it can be produced according to conventional methods by genetic engineering techniques. For example, levtin is described as an example as follows, but partial peptides can also be produced according to this. First, the gene coding for levutin (the levulin gene) is converted into a conventional genetic engineering method (eg, Sambrook J., Frisch EF, Maniatis T., Molecular Cloning 2nd edition). Acquired in accordance with Cold Spring Harbor Laboratory Press). The leptin gene is preferably a human leptin gene, and its DNA sequence including a sequence encoding a signal peptide is as shown in SEQ ID NO: 3. Then, using the obtained levtin gene, levtin can be produced according to a conventional genetic engineering method.

[0015] For example, by preparing a plasmid that allows the lebutin gene to be expressed in a host cell, introducing the plasmid into the host cell, transforming it, and then culturing the transformed host cell (transformant). What is necessary is just to acquire levtin from the obtained culture.

[0016] The above plasmid contains, for example, genetic information that can be replicated in the host cell, can be propagated autonomously, can be easily isolated and purified from the host cell force, and functions in the host cell. Preferable examples include those in which a gene encoding lebutin is introduced into an expression vector having a detectable promoter and a detectable marker. For example, an expression vector used for expression in Escherichia coli is an expression vector containing a promoter such as lac, trp, tac and the like, and these are commercially available from Falmacia, Takara Shuzo and the like. If it is necessary to induce further high expression, the gene is encoded upstream of the gene encoding levutin. Bosome binding regions may be linked. Examples of the ribosome binding region used include those described in reports by Guarente L. et al. (Cell 20, p543) and Taniguchi et al. (Genetics of Industrial Microorganisms, p202, Kodansha).

[0017] Examples of host cells include prokaryotic or eukaryotic microbial cells, insect cells, or mammalian cells. For example, Escherichia coli and the like can be preferably mentioned from the viewpoint of facilitating large-scale preparation of lebutin.

[0018] The therapeutic agent of the present invention exhibits an excellent effect in the treatment of resting alopecia. Resting alopecia to which the therapeutic agent of the present invention can be applied includes androgenetic alopecia, postpartum (postpartum) alopecia, alopecia after taking pills, diet, starvation alopecia, pressure alopecia, hypothyroidism Alopecia due to pituitary function, alopecia due to hypoparathyroidism, drug-induced alopecia, seborrheic alopecia, traction alopecia, erosion alopecia, alopecia associated with endocrine disease, Examples include alopecia due to contact dermatitis, and male pattern alopecia is particularly preferable. When the therapeutic agent for resting alopecia of the present invention is locally administered externally, it is preferably applied locally or injected subcutaneously. The dose of levbutin or its partial peptide, which is the active ingredient, varies depending on the subject's gender, age, weight, and condition (symptoms). When applied locally, it should be selected appropriately according to the skin area to be administered. However, the area of the application site is usually about 10 cm ² , and it is usually about 10 to L000 mg, preferably about 10 to 500 mg per day. The above dose should be applied once or several times a day. In the case of subcutaneous injection, the dose of levtin or a partial peptide thereof can be appropriately selected depending on the skin area to be administered, but is usually about 1 to about 10 cm ² of the area of application site: L000 mg, preferably about 10-100 mg. The frequency of administration is not particularly limited, but it is preferably administered once a day to once every several weeks.

[0019] The therapeutic agent for alopecia of the present invention is locally administered. Examples of dosage forms include semi-solid preparations such as ointments and creams, liquid preparations such as shampoos, rinses and lotions, and tapes when applied locally, and injections when administered subcutaneously locally. Drugs are listed. In principle, the formulation method for these dosage forms follows the usual method. However, when these formulations are produced, an appropriate amount of the dosage form according to the formulation form is assumed. Body or additive can be used.

For example, in the case of an injection, in addition to a solvent (eg, purified water), a dispersant (eg, Tween 80 (manufactured by Atlas Powder Co., USA), HCO60 (manufactured by Nikko Chemicals), polyethylene glycol, etc. , Carboxymethylcellulose, sodium alginate, etc.), preservatives (methylparaben, propylparaben, benzyl alcohol, etc.), isotonic agents (salt sodium, man-tol, sorbitol, glucose, etc.) can do.

On the other hand, in the case of preparations suitable for topical application (for example, ointments, creams, lotions, etc.) It is preferable to make the partial peptide easy to reach the hair follicle tissue.

[0020] Ribosomeization means inclusion of a drug in the interior of a ribosome (that is, a closed vesicle composed of a membrane-assembled lipid and an internal aqueous phase), and levtin or a partial peptide thereof. May be contained in a lipid layer assembled in a membrane form constituting a ribosome, or may be contained in an aqueous phase inside the ribosome.

[0021] In the ribosome according to the present invention, various lipophilic or hydrophilic components can be encapsulated in the lipid membrane and the inner aqueous phase, respectively.

[0022] Ribosomeization of lebutin or a partial peptide thereof can be performed by a conventional method. For example, lebutin or its partial peptide can be dissolved in a membrane constituent lipid component, mixed with a membrane constituent lipid component in a water-soluble non-volatile organic solvent, dissolved or suspended in an aqueous solution, etc. It can be encapsulated in a lipid membrane or an inner aqueous phase. The therapeutic agent for resting alopecia is obtained by dispersing the ribosomal levtin prepared as described above or a partial peptide thereof in a therapeutic agent base.

[0023] Examples of the lipid used for ribosome formation include glyceport phospholipid, glyceport glycolipid, sphingophospholipid, sphingoglycolipid, sterols and the like. Examples of the dalycerophospholipid include phosphatidylcholine, phosphatidylglycerol, phosphatidylinositol, and phosphatidylserine. Examples of the glyceglycoglycolipid include digalatatosyl diglyceride and galactosyl diglyceride. Examples include phingomyelin, glycosphingolipids include celebrity mouthsides and gandariosides, and sterols include cholesterol and cholesterol hemisuccinate. Here, phosphatidylcholine, phosphatidylglycerol, phosphatidylinositol, phosphatidylserine, sphingomyelin, ganglioside, etc. each have two saturated or unsaturated fatty acid ester chains, but the fatty acid (alkanoyl or alkenoyl group) moiety carbon Lipids having a number between 10 and 18 can be used as liposome constituents. Examples of the alkanoyl or alkenoyl group having 10 to 18 carbon atoms include desilyl, undesilyl, lauroyl, tridesilyl, myristoyl, pentadesilyl, noremitoyl, heptadesilyl, stearoyl, and oleoyl groups. Preferably, lauroyl, Myristoyl, palmitoyl and stearoyl groups. The aqueous phase that forms the interior of the ribosome is usually an aqueous sodium chloride solution, buffer solution (phosphate buffer, acetate buffer, etc.), monosaccharide or disaccharide aqueous solution (dulcose aqueous solution, trehalose aqueous solution, etc.). Can be used.

In order to obtain a preparation suitable for topical application using the ribosomal levtin or its partial peptide obtained as described above, it may be carried out according to a method employed in this field.

Example

[0024] The ability to explain the present invention using test examples below The present invention is not limited to this.

[0025] [Test Example 1]

(1) Examination of hair cycle in db / db (leptin receptor deficient) mice

35-day-old C57BLZKsJ—dbZdb mice (leptin receptor-deficient mice) obtained from Nippon Claire Co., Ltd. and normal mice of the same strain (n = 3 each) were excised and fixed with formalin. Hematoxylin and eosin stained specimens were prepared and the state of hair follicles was observed. As a result, in all individuals, the growing hair follicle was recognized in the normal mouse (B in FIG. 1), and only the resting hair follicle was observed in the 1S dbZdb mouse (C in FIG. 1). This confirms that dbZdb mice cannot enter the second growth phase. Fig. 1A shows the normal hair cycle of mice. [0026] (2) Growth phase induction by hair removal in ob / ob (leptin deficient) mice

7 days old (second resting) C57BLZ6J—obZob mouse leptin-deficient mice) obtained from Nippon Claire Co., Ltd. and normal mice of the same strain (n = 3 each) were removed, and the dorsal skin 14 days later After fixation with formalin, a methoxylin-eosin stained specimen was prepared and the condition of the hair follicle was observed. As a result, in all individuals, normal mice showed follicular hair follicles (Fig. 2B), but obZob mice showed only resting hair follicles (D in Fig. 2). From this, it was verified that obZob mice cannot induce the growth phase by hair removal stimulation. In addition, A and C in Fig. 2 are photographs of the back of the mouse immediately before the dorsal skin resection.

[0027] (3) Growth phase induction by leptin in ob / ob (leptin deficient) mice

The above-mentioned 7-week-old (second resting) obZob mice (n = 3) were removed from their back hairs, and a saline solution (10 μg / 0.lm 1) of mouse recombinant leptin (R & D Systems) was used. After a single subcutaneous injection, on the 16th day, the dorsal skin turned black, confirming that the growth phase had been induced (B in Fig. 3). In the control group to which only physiological saline was administered, the back skin was hardly changed (A in Fig. 3). From this, it was verified that levtin induces the growth phase from the resting phase in individuals with levutin deficiency.

[0028] (4) Growth phase induction by leptin in C57BLZ6J mice (normal mice)

The above-mentioned 7-week-old (second resting) C57BLZ6J mouse (normal mouse, n = 3) is shaved on the back hair, and the mouse recombinant leptin saline solution (10 gZO.lml) or saline only When mice were injected subcutaneously with lebutin, hair growth was observed on the 21st day in line with the subcutaneous injection site (C, arrow in Fig. 4), and clearly histologically grown. Although it was confirmed to be a hair follicle (Fig. 4, B, arrow), no hair growth was observed in mice subcutaneously injected with physiological saline (C, lower portion of Fig. 4), and histologically also paused. (Figure 4A). This proved that leptin induces the growth phase from the resting phase even in normal individuals.

[0029] [Test Example 2]

Growth phase induction by partial peptides in C57BLZ6J mice (normal mice)

The above-mentioned 7-week-old (second resting) C57BLZ6J mouse (normal mouse, n = 3) was shaved from the back hair, and a partial peptide of human leptin (22-56 residues, SEQ ID NO: 4, BRACHEM A single subcutaneous injection of physiological saline solution (10 / z gZO. 1 ml) or saline alone, and in mice injected with a partial peptide, it coincided with the subcutaneous injection site on the 21st day. (B in Fig. 5), mice with subcutaneous injection of saline showed no hair growth (A in Fig. 5). From this, it was verified that even in normal individuals, partial peptides induce growth from the resting phase.

[0030] (5) Consideration

From the above experiments, it was proved that levtin or its partial peptide has a therapeutic effect on resting alopecia.

Industrial applicability

[0031] The therapeutic agent for resting alopecia according to the present invention can be used as a medicament for the treatment of resting alopecia.

Claims

The scope of the claims

[I] A therapeutic agent for resting alopecia, comprising levtin or a partial peptide thereof as an active ingredient.

[2] The therapeutic agent according to claim 1, wherein the active ingredient is levutin.

[3] The therapeutic agent according to claim 1, wherein the active ingredient is a partial peptide of lebutin.

[4] The therapeutic agent according to claim 2, wherein the lebutin is human leptin.

[5] The therapeutic agent according to claim 4, wherein the human leptin is human recombinant levbutin.

[6] The therapeutic agent according to claim 3, wherein the partial peptide is represented by SEQ ID NO: 4.

[7] The therapeutic agent according to any one of claims 1 to 6, wherein the resting alopecia is androgenetic alopecia.

[8] The therapeutic agent according to any one of claims 1 to 7, wherein the formulation type is an injection for subcutaneous administration.

[9] The therapeutic agent according to any one of claims 1 to 7, wherein the formulation type is a formulation for topical application.

[10] The therapeutic agent according to claim 9, wherein the active ingredient is ribosomeized!

[II] Use of lebutin or a partial peptide thereof for producing a therapeutic agent for resting alopecia

[12] A method for treating resting alopecia characterized by administering a therapeutically effective amount of levutin or a partial peptide thereof to a patient with resting alopecia.