KR20210076494A - Composition for preventing hair loss or promoting hair growth comprising thrombopoietin - Google Patents

Abstract

The present invention relates to a composition for preventing hair loss or promoting hair growth comprising thrombopoietin. In the present invention, it is confirmed that a hair growth effect in mice is significantly increased by thrombopoietin, and it is confirmed that the growth of mouse mustache hair follicles is promoted, and dermal papilla cells are stimulated. Thus, thrombopoietin of the present invention can be easily utilized for preventing hair loss or promoting hair growth.

Description

A composition for preventing hair loss or promoting hair growth comprising thrombopoietin {Composition for preventing hair loss or promoting hair growth comprising thrombopoietin}

The present invention relates to a composition for preventing hair loss or promoting hair growth comprising thrombopoietin.

Recently, as interest in beauty increases, interest in the treatment of alopecia is also increasing. Alopecia refers to a condition in which there is no hair where it should normally be. Although hair does not have an important physiological function directly related to life, it plays a very large role from a cosmetic point of view. In addition, it has functions such as UV protection and hair protection. If hair loss is severe, there may be problems in social life, and it can seriously affect psychologically, so it is important in terms of quality of life.

Hair loss can be clinically divided into scarring alopecia accompanied by wounds and non-scarring alopecia in which only hair falls out. In the case of scarring alopecia, hair follicles are destroyed, so hair does not grow back. Hair is made in places called hair follicles, and each hair follicle goes through a cycle of activity and rest. The time interval of this hair cycle varies depending on the body part. In the case of hair, it goes through an anagen phase of 2 to 6 years and a catagen phase of 2 to 4 weeks before entering a telogen of 3 to 4 months. do. Each hair follicle has 10 to 20 hair follicle growth cycles in its lifetime.

The most widely used surgical method for hair loss treatment is autologous hair transplantation, and drug treatment using propecia and minoxidil is widely used. In the case of drug treatment, hair growth effect appears at the time of administration, but hair loss proceeds again when treatment is stopped. In particular, in the case of minoxidil, there are side effects such as sexual dysfunction.

On the other hand, thrombopoietin (TPO or THPO) is a growth factor produced in the liver and kidney, and thrombopoietin acts through a thrombopoietin receptor to promote the maturation and differentiation of giant cells. create Thrombopoietin was mainly developed as a treatment for thrombocytopenia, and there is no known content related to hair growth.

Therefore, in the present invention, as a result of earnest efforts to develop a more effective composition for preventing and treating hair loss, it was confirmed that hair growth induction was promoted by thrombopoietin, as well as that the hair growth effect was significantly increased in mice. and completed the present invention.

An object of the present invention is to provide a composition for preventing hair loss or promoting hair growth comprising thrombopoietin as an active ingredient.

In order to achieve the above object,

The present invention provides a composition for preventing hair loss or promoting hair growth comprising thrombopoietin as an active ingredient.

According to a preferred embodiment of the present invention, the composition may be a pharmaceutical composition, a quasi-drug composition or a cosmetic composition.

In the present invention, it was confirmed that the hair growth effect in mice was significantly increased by thrombopoietin, and it was confirmed that the growth of the mouse mustache hair follicle was promoted and that the dermal papilla cells were stimulated. Thrombopoietin can be easily utilized for the purpose of preventing hair loss or promoting hair growth.

1 is data showing the results of measuring hair growth and hair weight after injection of thrombopoietin into hair-removed mice.
Figure 2 is data confirming the change in the length of the mustache due to thrombopoietin treatment in the mustache hair follicles of a mouse.
3 is data confirming the degree of proliferation of dermal papilla cells (hDPC) by thrombopoietin.
4 is data confirming the change in dermal papilla cell marker gene expression by thrombopoietin.

Hereinafter, the present invention will be described in detail.

The present invention, from one point of view, relates to a composition for preventing hair loss or promoting hair growth comprising thrombopoietin as an active ingredient.

The thrombopoietin (TPO or THPO) is a growth factor produced in the liver and kidney, and thrombopoietin acts through a thrombopoietin receptor to promote the maturation and differentiation of giant cells, create Thrombopoietin was mainly developed as a treatment for thrombocytopenia, and there is no known content related to hair growth.

In the present invention, thrombopoietin containing the amino acid represented by SEQ ID NO: 1 was preferably used, but recombinant thrombopoietin or thrombopoietin analog having an effect of preventing hair loss or promoting hair growth may be used without limitation.

"Hair loss" that can be prevented or treated by the composition of the present invention is a phenomenon in which hair comes out completely out of the scalp, and it can prevent or treat male pattern hair loss and female pattern hair loss, and prevention or treatment of male pattern hair loss caused by hair follicle damage is preferred, but is not limited thereto.

As used herein, the term “hair growth” refers to hair growth from the scalp, and specifically refers to inducing hair growth by forming hair follicles in a bald area or a hairless area (hairless area). It is used synonymously with "hair growth" or "wool" in the art to mean lengthening of hair (ie, hair growth).

In a specific embodiment of the present invention, as a result of injecting thrombopoietin into the back of a hair-removed mouse, as shown in FIG. 1 , it was confirmed that the anagen induction effect was excellent.

In addition, as a result of treating the mouse mustache hair follicle with thrombopoietin, as shown in FIG. 2 , it was confirmed that the length of the mouse mustache was significantly increased.

In another specific embodiment of the present invention, as a result of treating dermal papilla cells with thrombopoietin, as shown in FIG. 3, it was confirmed that the proliferation of dermal papilla cells significantly increased in a thrombopoietin concentration-dependent manner, FIG. As shown in Fig. 4, it was confirmed that thrombopoietin increased the expression of the mouse fodder cell marker gene.

Therefore, since it was confirmed that the thrombopoietin of the present invention effectively induces hair growth, it can be easily utilized for preventing hair loss or promoting hair growth.

The composition of the present invention may be prepared as a pharmaceutical composition, a quasi-drug composition or a cosmetic composition.

When the composition of the present invention is prepared as a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are commonly used in formulation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. doesn't happen The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like, in addition to the above components. Suitable pharmaceutically acceptable carriers and agents are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, mucosal administration, and eye drop administration.

A suitable dosage of the pharmaceutical composition of the present invention is variously prescribed depending on factors such as formulation method, administration method, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate and reaction sensitivity of the patient. can be Preferably, the dosage of the pharmaceutical composition of the present invention is 0.0001-100 mg/kg (body weight) based on an adult.

The pharmaceutical composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person of ordinary skill in the art to which the present invention pertains. or may be prepared by incorporation into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension, syrup, or emulsion in oil or aqueous medium, or may be in the form of an extract, powder, powder, granule, tablet or capsule, and may additionally include a dispersant or stabilizer.

When the composition of the present invention is prepared as a quasi-drug composition, the adipose-derived stem cells exhibiting the effect of preventing hair loss or promoting hair growth may be added as it is, or may be used together with other quasi-drugs or quasi-drug ingredients, and may be appropriately used according to a conventional method. have. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use.

The quasi-drug composition for preventing hair loss or promoting hair growth of the present invention is not particularly limited in its formulation, and it can be formulated in various forms in the form of quasi-drugs known in the art as exhibiting an effect of preventing hair loss or promoting hair growth. The formulated quasi-drugs are hair tonic, hair lotion, hair cream, hair spray, hair mousse, hair gel, hair conditioner, hair shampoo, hair conditioner, hair pack, hair treatment, eyebrow hair growth agent, eyelash hair growth agent, eyelash nutrient agent, pet. There are shampoo for pets, rinse for pets, hand sanitizer, detergent soap, soap, disinfectant, wet tissue, mask, ointment, patch or filter filler, etc., and includes all quasi-drugs in the ordinary sense.

In addition, in each formulation, the quasi-drug composition for preventing hair loss or promoting hair growth can be formulated by selecting other ingredients arbitrarily according to the formulation or purpose of use of other quasi-drugs. The mixing amount of the active ingredient may be suitably determined depending on the intended use, and may include, for example, conventional auxiliary agents such as thickeners, stabilizers, solubilizers, vitamins, pigments and fragrances, and carriers.

When the composition of the present invention is prepared as a cosmetic composition, the ingredients included in the cosmetic composition of the present invention include ingredients commonly used in cosmetic compositions in addition to the adipose-derived stem cells as an active ingredient, such as antioxidants, stabilizers, conventional adjuvants such as solubilizers, vitamins, pigments and fragrances, and carriers.

The cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing , oil, powder foundation, emulsion foundation, wax foundation, spray, etc., but is not limited thereto.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane /may contain propellants such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylglycol oil, glycerol fatty esters, fatty acid esters of polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystals Adult cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used.

When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide as carrier components Ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative or ethoxylated glycerol fatty acid ester and the like can be used.

Hereinafter, the present invention will be described in more detail through examples.

These examples are only for illustrating the present invention, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not to be construed as being limited by these examples.

thrombopoietin Confirmation of hair growth effect by treatment

Ten 6-week-old male C3H/HeN mice were shaved short using a depilatory machine, and the remaining hair was completely removed using a depilatory agent, and then stabilized for one day before use in the experiment.

The experiment was performed by dividing 4 animals in each group, and the groups are as follows.

Group 1: Control

Group 2: Thrombopoietin 100 ng group

Each group was subcutaneously injected three times on the back of the mouse on 3 days, and the degree of hair growth in the back of the mouse was tracked for 16 days after injection of thrombopoietin (peptoTech, cat No. 300-18, SEQ ID NO: 1). observed. After 16 days, after shaving the back of the mouse with hair growth using a razor, hairs were obtained and weight was measured.

As a result, as shown in FIG. 1 , it was confirmed that the thrombopoietin-administered group had better anagen induction effect than the control group.

thrombopoietin Confirmation of hair growth induction effect according to treatment

In the present invention, it was attempted to confirm the effect of thrombopoietin treatment on hair growth induction.

Specifically, the anagen vibrissal of C57/BL6 female mice was separated using a scalpel and tweezers. The mustaches of the isolated mice were added to a quantitative medium (2 mM L-glutamine, 10 μg/ml insulin, 10 ng/ml hydrocortisone, penicillin and serum-free Williams E medium) with thrombopoietin at 5 ng and 20 ng, respectively. treated and cultured. Individual hair follicles were photographed 72 hours after initiation of culture, and the change in hair length was calculated from the photos and expressed as the mean ± SE of 8-10 hair follicles.

As a result, as shown in FIG. 2 , in the group treated with thrombopoietin than in the control group, the length of the mustache of the separated mice was increased.

thrombopoietin Confirmation of dermal papilla cell proliferation effect according to treatment

Human dermal papilla cell (hDPC) in 1% antibiotic and antimycotic (Hyclone) containing dermal papilla cell growth media (Follicle Dermal Papilla cell Growth media, Promocell, Germany) Cells were prepared by inoculating the medium and culturing in a 5% carbon dioxide incubator at 37°C. Then, the hDPC cultured above in a 12-well plate was ^{inoculated into each well at a concentration of 5×10 3} cells/well, incubated overnight, and then treated with 5 ng and 20 ng of thrombopoietin for 48 hours. during further incubation.

Thereafter, 500 μl of trypsin was added to each well to separate the cells, and then the number of cells was counted to determine the degree of cell proliferation.

As a result, as shown in FIG. 3 , it was confirmed that the proliferation of dermal papilla cells was increased when thrombopoietin was treated.

thrombopoietin Confirmation of expression level of dermal papilla cell markers according to treatment

RNA was isolated from dermal papilla cells cultured by treatment with 20 ng of thrombopoietin of Example 3 to confirm the expression level of dermal papilla cell markers.

Specifically, total RNA was isolated and quantified from dermal papilla cells using Trizol, and 500ng of the quantified total RNA was used to contain 50ng/μl of oligo (dT) primer, 10mM dNTP, and 10,000U of reverse transcriptase. cDNA was synthesized with The expression levels of ALP (alkaline phosphatase), VCAN, CORIN, LEF1, NESTIN, and ACTA1, known as dermal papilla cell markers, were analyzed using the cDNA and the primers of Table 1 below using real-time QPCR (quantitative-PCR).

Primer information for performing PCR primer sequence SEQ ID NO: ALP F: 5'-GATCCTAAAAGGGCAGAAGA-3' SEQ ID NO: 2 R: 5'-GGCACATGCTTGTCTACACT-3' SEQ ID NO: 3 VCAN F: 5'-GTAACCCATGCGCTACATAAAGT-3' SEQ ID NO: 4 R: 5'-GGCAAAGTAGGCATCGTTGAAA-3' SEQ ID NO: 5 CORIN F: 5'-CCAAAGCCGGTCTTGAGAG-3' SEQ ID NO: 6 R: 5'-GAGGAGGTTAGCAGTCGCC-3' SEQ ID NO: 7 LEF1 F: 5'-GTCCACACATCCTCCAGC-3' SEQ ID NO: 8 R: 5'-GTGTCGACTGACAGTGAGG-3' SEQ ID NO: 9 NESTIN F: 5'-TCTGGAAGAGGAAGACAACC-3' SEQ ID NO: 10 R: 5'-GTCCTTCTCCACCCGTATCTT-3' SEQ ID NO: 11 ACTA1 F: 5'-CTCACCGACTACCTGATGAA-3' SEQ ID NO: 12 R: 5'-GAGCACAGCTTCTCCTTCA-3' SEQ ID NO: 13 GAPDH F: 5'-CGAGATCCCTCCAAAATCAA-3' SEQ ID NO: 14 R: 5'-TGTGGTCATGAGTCCTTCCA-3' SEQ ID NO: 15

As a result, as shown in FIG. 3 , it was confirmed that the expression of dermal papilla cell markers was increased by thrombopoietin, and in particular, it was confirmed that the expression of ALP, VCAN, NESTIN and ACTA1 genes was significantly increased. That is, it was confirmed that the dermal papilla cells were stimulated by thrombopoietin.

<110> Industry-Academic Cooperation Foundation, Yonsei University <120> Composition for preventing hair loss or promoting hair growth comprising thrombopoietin <130> 1066645 <160> 15 <170> KoPatentIn 3.0 <210> 1 <211> 174 <212> PRT <213> Artificial Sequence <220> <223> Recombinant Human TPO <400> 1 Ser Pro Ala Pro Pro Ala Cys Asp Leu Arg Val Leu Ser Lys Leu Leu 1 5 10 15 Arg Asp Ser His Val Leu His Ser Arg Leu Ser Gln Cys Pro Glu Val 20 25 30 His Pro Leu Pro Thr Pro Val Leu Leu Pro Ala Val Asp Phe Ser Leu 35 40 45 Gly Glu Trp Lys Thr Gln Met Glu Glu Thr Lys Ala Gln Asp Ile Leu 50 55 60 Gly Ala Val Thr Leu Leu Leu Glu Gly Val Met Ala Ala Arg Gly Gln 65 70 75 80 Leu Gly Pro Thr Cys Leu Ser Ser Leu Leu Gly Gln Leu Ser Gly Gln 85 90 95 Val Arg Leu Leu Leu Gly Ala Leu Gln Ser Leu Leu Gly Thr Gln Leu 100 105 110 Pro Pro Gln Gly Arg Thr Thr Ala His Lys Asp Pro Asn Ala Ile Phe 115 120 125 Leu Ser Phe Gln His Leu Leu Arg Gly Lys Val Arg Phe Leu Met Leu 130 135 140 Val Gly Gly Ser Thr Leu Cys Val Arg Arg Ala Pro Pro Thr Thr Ala 145 150 155 160 Val Pro Ser Arg Thr Ser Leu Val Leu Thr Leu Asn Glu Leu 165 170 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> ALP_F <400> 2 gatcctaaaa gggcagaaga 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> ALP_R <400> 3 ggcacatgct tgtctacact 20 <210> 4 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> VCAN_F <400> 4 gtaacccatg cgctacataa agt 23 <210> 5 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> VCAN_R <400> 5 ggcaaagtag gcatcgttga aa 22 <210> 6 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> CORIN_F <400> 6 ccaaagccgg tcttgagag 19 <210> 7 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> CORIN_R <400> 7 gaggaggtta gcagtcgcc 19 <210> 8 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> LEF1_F <400> 8 gtccacacat cctccagc 18 <210> 9 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> LEF1_R <400> 9 gtgtcgactg acagtgagg 19 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> NESTIN_F <400> 10 tctggaagag gaagacaacc 20 <210> 11 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> NESTIN_R <400> 11 gtccttctcc acccgtatct t 21 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> ACTA1_F <400> 12 ctcaccgact acctgatgaa 20 <210> 13 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> ACTA1_R <400> 13 gagcacagct tctccttca 19 <210> 14 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> GAPDH_F <400> 14 cgagatccct ccaaaatcaa 20 <210> 15 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> GAPDH_R <400> 15 tgtggtcatg agtccttcca 20

Claims (4)

A composition for preventing hair loss or promoting hair growth comprising thrombopoietin as an active ingredient.
The composition for preventing hair loss or promoting hair growth according to claim 1, wherein the composition is a pharmaceutical composition.
The composition for preventing hair loss or promoting hair growth according to claim 1, wherein the composition is a quasi-drug composition.
The composition for preventing hair loss or promoting hair growth according to claim 1, wherein the composition is a cosmetic composition.

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