Indian Patents. 222068:"A HAIR GROWTH PROMOTING AGENT COMPRISING A 3-POSITION ANALOG OF CYCLOSPORINE "

Title of Invention	"A HAIR GROWTH PROMOTING AGENT COMPRISING A 3-POSITION ANALOG OF CYCLOSPORINE "
Abstract	he present invention discloses a hair growth promoting agent comprising a cyclosporin derivatives as an active ingredient, and more particularly, a hair growth promoting agent comprising a cyclosporin. A derivative substituted in the 3-position as an active ingredient.

Full Text	USE OF 3-POSITION CYCLOSPORIN DERTVATI\"ES FOR HAIR GROWTH Technical Field The present invention relates to a hair growth promoting agent comprising a cyclosporin derivative as an active ingredient and more particularly, to a hair growth promoting agent comprising cyclosporin derivatives modified in the 3-position as an active ingredient. Background Art On average, the human scalp contains about 100,000 to 150,000 hairs. Each hair has three main stages of growth: anagen, catagen and telogen, after which the hair falls out. This hair growth cycle is repetitive and the duration of one cycle is different from other cycles, ranging approximately 3 to 6 years. Thus, the average adult normally loses about 50 to 100 hairs every day. In general, alopecia refers to a phenomenon wherein duration of the anagen growth phase is shortened and the percentage of hairs in the catagen and telogen phases increases, whereby the number of lost hairs is increased excessively and abnormally. There are many theories to explain for loss of hair, including for example, poor blood circulation, excessive functioning of male sex hormone, excessive production and secretion of sebum, deterioration of scalp by peroxides, bacteria, etc., hereditary factors, aging, stress, etc. However, explicit mechanisms have not been revealed. Recently, the population suffering from hair loss is tending to increase, since changing dietary habits and stress imposed on individuals due to modern social environments, etc. has increased. Also, the age of the individuals affected by alopecia is dropping and furthermore, the population of female alopecia sufferers is rising. One of preparations which are most commonly used for treatment and M* prevention of alopeciais one that contains minoxidil. There are two hair-regrowth agents which have received approval from the U.S. Food and Drug Administration, and minoxidil is one of those approved hair-regrowth agents. Minoxidil was originally developed as a hypertension drug for the purpose of reducing blood pressure. However, when using this drug, as a side effect, a trichogenous effect was observed and thereafter, this drug became famous as a hair-regrowth agent. Although mechanisms by which minoxidil works as a hr.r-regrowth asent is no: clearly understood, it is inferred that minoxidil increases blood flow by expansion of blood vessels, whereby roots of hairs are supplied with more nutrition and eventually, growth of hairs are promoted, Such a model of bicod flow increase has been indirectly supported by a recent report that minoxidil enhances the expression of vascular endothelial growth factor (VEGF), a growth factor associated with vasodiiata:ion in the dermal papilla which is a main cell making up the hair roots. Also, other than the vasodilative effect of the minoxidil in the hair-restoring mechanism, it has been reported that minoxidil promotes activation of dermal papilla cells in the roots of hair incubated in vitro, and growth of hair follicles in a tissue culture of follicles in vitro. These facts indicate that minoxidii may work directly on the roots of hair as a growth factor. In addition, finasteride, a main component of Propecia which has started to be sold by Merck, is used for treatment of alopecia. It inhibits conversion of the male hormone testosterone into dihydrotestosterone, which is a more potent male hormone than testosterone, On December of 1997, the 1 mg finasteride tablet was approved by the US FDA as a hair-regrowth agent for treatment of male pattern hair loss in men only, and is now commercially available. In clinical studies, it has been demonstrated to have a significant trichogenous effect. However, there has been a report that finasteride may inhibit male sexual function as a side effect. Since neither finasteride nor minoxidil show superior effect in clinical tests, and there is concern about side effects, many researches are conducted to develop a new and improved hair-regrowth agents. The cyclosporin family of drugs has immunosuppressive activity. It is also effective to inhibit growth of virus, fungus, protozoan, etc. and has various physiological effects such as nephrotoxicity, hepatotoxicity, hypertension, enlargement of periodontium, trichogenous effect, and so on, as side effects. Cyclosporin A, a representative cyclosporin, is a cyclic peptide having the following Chemical Formula, "which comprises 11 amino acids, including several N-methyl araino acids and D-alanine at No. 8 residue. [Structure Formula 1] pMeBmt-Abu-Sar-MeLeu—Val-MeLeu-Aia-DA!a-MeLeu —MeLeu— 1 23 4 5,6 78 5 10 11 where MeBmt is N-raethyl-(4R)-4-"k(E)-2-biitenyrj-4-methyl-L--Jireonhe, Abu is L-a-aminobutyric acid, Sar is sarco sine, MeLeu is N-methyl-L-leucir.e,Val is L-valine, Ala is L-alanine, DAla is D-alanine, MeVal is N-raethyl-L-valine. The amino acid form of cyclosporin A of the above Chemical Formula 1 is L-configuration, unless otherwise specified. The residue numbering of amino acids starts from MeBmt and proceeds clockwise, i.e. 1 for MeBmt and 11 for the last MeVal (N-methyl-L-valine) as shown in the Structure Formula 1. Nomenclature of '.•arious derivatives including cyclosporins A to Z, follows methods commonly used (Helv. Chim. Acta, 1987, 70:13-36). For example, if Abu in the 2-position of cyclosporin A is substituted with L-alanine, L-threonine, L-valine or L-norvaline, the derivatives thus prepared are named cyclosporin B, cyclosporin C, cyclosporin D or cyclosporin G, respectively. Further, when the amino acid residues of the cyclosporin derivatives differ from those of cydcsporin A, the derivatives are named by describing the substituent For example, if sarcosine, being the amino acid residue 3 of cyclosporin A, is substituted with N-methyl-D-Abu3 or N-methyl-D-Nva3, the derivatives thus prepared are named [N-methyl-D-Abu3] cyclosporin A or [N-methyl-D-Nva3] cyclosporin A, respectively. Meanwhile, a common method for abbreviating amino acids is employed, that is, N-methyl-L-leucine is abbreviated by MeLeu, N-methyl-L-isoleucine by MeUe, N-methyl-L-Valine by MeVal, N-methyl-L-alanine by MeAla, N-rr.ethyl-L-norvaline by MeNva, L-leucine by Leu, L-isoleucine by He, sarcosine by Sar, L-serine by Ser, L-valine, Val, L-alanine by Ala, D-alanine by DAla, L-aminobutviic acid by Abu, L-threonine by Thr, and L-norvaline by Nva. Further, as for a derivative of cyclosporin which is substituted with sulfur instead of a carbonyl oxygen at the amino acid residue 7, the name of the derivative may be cyclosporin 7-thioamide or [V CS-NH] cyclosporin, according to different references (Helv. Chint Acta. 74: 1953-1990,1991; J. Org, Chem. 58:673-677,1993; J. Org. Chem. 59:7249-7258,1994). So far, possible development of cyclosporin as a hair-regrowth agent has been studied by many research grouris. Particularly, researches involving animal hair reg^>wth tests, human alopecia artata (J. Am Acad. DerrnaioL, 1990,22^42-250), human male pattern alopecia (J. Am, Acad, DennatoL, 1990,22^51-253 and Skm PhanracoL, 1994,7:101-104), and inhibition effect of hair toss by chenwtherapy in animal models (Am. J. PathoL, 1997,150:1433-1441) have been widely conducted In comparative exrwimenrs on mouse's back, h is shown that cyclosporin has a hair regrowrh effect about 100 times superior to minoxidn Based on such findings, there have been attempts to utilize cyclosporin as a treatment for male pattern alopecia, and many applications for patents have been filed For example, Japanese Patent Publication Kokai Nos. Sho 60-243008, Sho 62-19512. and Sho 62-19513 disclose use of cyclosporin derivatives as a hair regrowth agent Also, Europe Patent Publication No. 0414632B1 teaches a cyclosporin derivative modified ir. the 8-position, and PCT Publication No. 93/1 7039 teaches isocyclosporin. Moreover, United States Patent No. 5,807,820 and British Patent No. 2,218,334A disclose cyclosporins with excellent transdermal absorption, pursuant to the use of cyclosporins as hair restorers. Disclosure of the Invention Therefore, the present invention has been made in view of the above problems associated with side effects of cyclosporin A, and it is an object of the present invention to provide a hair growth promoting agent comprising a cyclosporin derivative as an active ingredienti which exerts an excellent hair growth-promotion ability. In accordance with one aspect of the present invention, the above and other objects can be accomplished by the provision of a hair growth promoting agent comprising a 3-position analog of cyclosporin represented by the below Formula 1, as an active ingredient, which is prepared by synthesizing a variety of derivatives thereof and evaluating their hair growth promoting effects, with an aim of developing a novel agent for promoting hair growth, [Formula 1] A-B-C-D-E-F-G-H-I-J-K wherein: A represents N-me^l^RM-tCE^-butenylj-^methyl-L-threonine, (2S3R,4R,6E)-3-sulfhydryl4-mefeyl-2^methylarnino)-6-octenoic add or (2S,4R,6E)-3-oxo-4-methyl-2-(mrthylaraino>6-octenoic acid; B represents l^aminobutyric add (Abu), L-alanine (Ala), L-threonine (Thr), L-valine (Val) or L-ootvaline (Sva); C represents a D-amino add represented by the general formula 1, [General formula 1] in which, R is one selected from the group consisting of hydrogen, Q-Q straight or branched alkyl alkenyl or alkynyl moieties, subnituted or unsubstituted with one or more selectee from the group consisting of ammo, hydrox;., halo, haloaikyl, ester, alkoxy, cyano, ritro, alkylamino, and dialkylamino, and -X- R' represented by the general formula 2 below. [General formula 2] -X-R1 in which, X is oxygen or sulfur, and R1 is one selected from the group consisting of hydrogen, and C;-Cs straight or branched alkyl, alkenyl or alkynyl moieties, substituted or unsubsu'tuted with one or more selected from the group consisting of amino, hydroxy, halo, haloaikyl. ester, alkoxy, cyano, nitro, alkylamino, and dialkylamino; D represents N-methyl-L-leucine, y-hydroxy N-methyl-L-leucine or L-valine; E represents L-valine or L-norvaline; F represents N-methyl-L-leucine, y-hydroxy N-methyl-L-leucine or L-leucine; G represents L-alanine or L-alanine thioamide ([V CS-NH], NH-CHCH3-CS-); H represents a D-amino acid represented by the general formula 3, [General formula 3] -NH-CH(CH2R>COOH in which, R' is hydrogen or X-R represented by the general formula 4, [General formula 4] -X-R1 in which, X is oxygen or sulfur, and R' is one selected from the group consisting of hydrogen, and C'-Ce straight or branched alkyl, alkenyl or alkynyl moieties, substituted or unsubstituted with one or more selected from the group consisting of amino, hydroxy, halo, haloaikyl, ester, alkoxy, cyano, nitro, alkylamino, and dialkylamino; I represents N-methyi-L-leucine, Y-hydroxy-N-mcthyl-L-leucinetor L-leucine; J represents N-rnethyl-L-leucine, Y-hydroxy-N-rncthyl-L-leucine or L-leucine; and, . K represents N-methyl-L-valine or L-valine, In accordance with snotber aspect of the invention, that is provided a hair growth promoting agent comprising a 3-postion analog of cydosporin with an excellent hair growth promoting effect, represented by Formula 2 below, as an active ingredient. [Formula 2] MeBmt-A'-e'-C'-O'-E'-^'-G'-H'-l-MeVal wherein: MeBmt represents N-meihyl^4R)^[(E)-2-butenyl]4-rnethyl-L-threonine; A' represents L-arnip.obu:yric acid, L-alarJne, L-threonhe, L-valr.e or L-norvalir.e: B1 represents N-meuyl-D-arriinobur.Tic acid, X-methyi-D-norvaiie, D-2-;methylarruno)hexa-4-ynoyl, D-2-(methylamino)pent-4-ynoyl, D-2-methylthio-sarcosine, N-:nethyl-O-propenyl-D-serine or N-methyi-D-serine; C' represents N-metbyl-L-leucine, y-hydroxy-N-meth\i-L-!eucine orL-vaiine; D' represents L-valine or L-norvaline; E represents N-methyl-L4eucine, y-hydroxy N-methyl-L-leucine or L-leucine; F represents L-alanine or L-aianine thioamide ([7\(/S CS-NH], NH-CHCH:,-CS-); G' represents D-alanine or D-serine; H1 represents N-methyl-L-leucine, y-hydroxy N-methyl-L-leucine or L-leucine; I represents N-methyl-L-leucine, y-hydroxy N-methyl-L-leucine or L-leucine; and, MeVal represents N-methyl-L-valine. In accordance with another aspect of the invention, there is provided a hair growth promoting agent comprising a 3-position analog of cyclosporin with an excellent hair growth promoting effect, represented by Formula 3 below, as an active ingredient, [Formula 3] MeBmt-A"-8"-MeLeu-Val-MeLeu-Ala-DAIa-MeLeu-MeLeu-MeVa! wherein: 4 MeBmt represents N-metyK4R>4{(E>2-butenyi}4-methyl-L-threonine; A" represents Iralanine, L-threonine, L-valine or L-norvaline; B" represents N-norvaline, D-2-(medoylaraino)hcxa-4-YQoyi, I>2^methYlamino)pcnt-4-ynoyl, D-2-methylthio-sarcosiDc, N- MeLeu represents N-raethyi-L-leudnc; Val represents L-valine; Ala represents L-alanine; . DAla represents D-alanine; and, Me Val represents N-methyl-L-valine. In accordance with ye: another aspect of the present invention, there is provided a hair growth promoting agent, whose composition comprising a 3-position analog of cyclosporin may be formulated in the form of liquid formualtions, sprays, gels, pastes. emulsions, creams, conditioners or shampoos. Brief Description of the Drawings The above and other objects, features and other advantages of the present invention will be more clearly understood from the following detailed description taken in conjunction with the accompanying drawing, in which: Fig. 1 is a !H-NMR spectrum of [N-methyl-D-Abu3] cyclosporin A; Fig. 2 is a 13C-NMR spectrum of [N-methyl-D-Abu3] cyclosporin A; Fig. 3 is a 1H-NMR spectrum of [N-methyl-D-Nva3] cyclosporin A; Fig. 4 is a 13C-NMR spectrum of [N-methyl-D-Nva3] cyclosporin A; Fig. 5 is a !H-NMR spectrum of [D-2-(methylamino)hexa4-ynoyl3] cyclosporin A; Fig. 6 is a 13C-NMR spectrum of p-2-(methylamino)hexa-4-ynoylJ] cyclosporin A; Fig. 7 is a 1H-NMR spectrum of p-2-(methylamino)pent-4-ynoyl'>] cyclosporin A; Fig. 8 is a °C-NMR spectrum of p-2-(methylamino)pent-4-ynoyl3] cyclosporin A; Fig. 9 is a 'H-NMR spectrum of p^methylthioySar5] cyclosporin A; Fig. 10 is a 13C-NMR spectrum of [D-2-(methylthio)-Sar5] cyclosporin A; Fig. 11 is a 'H-NMR spectrum of [N-methyl-O-propenyl-D-Ser3] cyclosporin A; Fig. 12 is a 13C-NMR spectrum of [N-methyl-O-propenyl-D-Ser5] cyclosporin A; Fig. 13 is a 'H-NMR spectrum of [N-methyl-D-Ser3] cyclosporin A; and Fig. 14 is a I3ONMR spectrum of [N-methyl-D-Scr3] cyclosporin A. 4 Best Mode far Carrying Out the Invention Hereinafter, the present invention wffl be described in detail, in conjunction with various examples. These examples arc provided only for illustrative purposes, and the present invention ts not to be construed as being limited to those examples,. With the aim of developing a novel agent with hair growth promoting effect, the present inventors chemically synthesized a variety of 3-position analogs of cyclosrorin, and hair growth promoting effects thereof were examined. Thus, the invention provides a hair growth promoting agent comprising a cyclosporin derivative as an active ingredient, Example 1: Synthesis of 3-position analog of cvclosporin A general method for the alkylation of cyclosporin A was as follows. Tetrahydrofuran (THF) was added with diisopropyl amine ((i-PrhNH) and added with a solution of n-butyl lithium (BuLi) in hexane under nitrogen atmosphere at -7S °C, followed by stirring for 30 mia To the solution of LDA (lithium diisopropylamide) thus prepared, cyclosporin A in THF was added, stirred for 1 hr, and electrophile was added. 1-1: Synthesis of fN-rneflivl-D-AbuJ] cvclosporin A: Compound 1 According to the general method above, to a solution of 10 equivalents of LDA was added 1.0 g cyclosporin A in 50 ml THF at -78 °C. The reaction mixture was stirred for 2 hrs at -78 °C and added with 0.4 ml ethyliodide. After the temperature of the solution reached room temperature, the solution was further stirred for 24 hrs and added with 20 ml water, followed by concentration. The residue was added with ether (EtiO), washed with water and a solution of saturated sodium chloride in sequence, and dried over anhydrous MgSO.;. After concentrating, the residue was subjected to silica gel column chromatography (100 g silica gel, dichloromethane : methylalcohol = 96 : 4), followed by HPLC to give 0.1 g of the title compound Molecular weight of the compound was determined by FAB MS (ZMS AX 505H) analysis. To confirm the molecular structure, Nuclear Magnetic Resonance (NMR) measurements were performed on 600 MHz (Bruker) for 'H-NMR and on 150 MHz (Bruker) for 13C-NMR, and the spectra are shown in Figs. 1 and 2, respectively. j-2: Synthesis of fN-methvl-D-Nva3! cvclosporin A: Compound 2 4 According to the general method, to a solution of 10 equivalents of LDA was added 1.0 g cyclosporin A in-50 ml THF at -78 °C. The reaction mixture was stirred for 2 hrs at -78 °C and added with 0.41 ml propylkxlide. After the temperature of the solution reached room temperature, the solution was further stirred for 24 hrs and added with 20 ml water, followed by ccocentratioa The residue was added with ether (EtjOX washed with water and a solution of saturated sodium chloride in sequence, and dried over-anhydrous MgS04. After concentrating, the residue was subjected to silica gel column chromatography (: 00 g silica gel, dichloromethane : methylalcohol = 96 : 4), followed by HPLC to give 0.12 g of the title compound. Molecular weight of the compound was determined by FAB MS (ZMS AX 505H) analysis, To confirm the molecular structure, Nuclear Magnetic Resonance (NMR) measurements were performec on 600 MHz (Bruker) for 'H-NMR and on 150 :MHz (Bruker) for 13C-NMR, and the spectra are shown in Figs. 3 and 4, respectively. 1-3: Synthesis P-2-(methvlarnmo)hexfl-4-vnovl3'l cvclosporin A: Compound 3 According to the general method, to a solution of 10 equivalents of IDA was added 1.0 g cyclosporin A in 50 ml THF at -78 °C. The reaction mixture was stirred for 2 hrs at -78 °C and added with 0.73 ml l-bromo-2-butyne. After the temperature of the solution reached room temperature, the solution was further stirred for 24 hrs and added with 20 ml water, followed by concentration. The residue was added with ether (Et20), washed with water and a solution of saturated sodium chloride in sequence, and dried over anhydrous MgSO. After concentrating, the residue was subjected to silica gel column chromatography (100 g silica gel, dichlororaethane: methylalcohol = 96:4), followed by HPLC to give 0.13 g of the title compound Molecular weight of the compound was determined by FAB MS (ZMS AX 505H) analysis. To confirm the molecular structure, Nuclear Magnetic Resonance (NMR) measurements were performed on 600 MHz (Bruker) for 1H-NMR and on 150 MHz (Bruker) for 13C-NMR, and the spectra are shown in Figs. 5 and 6, respectively. 1-4: Synthesis of p-24methylamino)pent-4-ynovlJl cvclosporin A: Compound 4 According to the general method, alkylation was performed employing THF (200 ml), (i-Pr):NH (3.2 ml), BuLi (8 ml), cyclosporin A (3.76 g) in 50 ml THF and propargyl bromide (3.57 g). After the temperature of the solution reached room temperature, the solution was further stirred for 24 hrs and added with 40 ml water, followed by concentration. The residue was added with ether (EtaO), washed with water and a solution of saturated sodium chloride in sequence, and dried over anhydrous MgSCu. After concentrating, the residue was subjected to silica gel column chromatography (100 g silica gel, dichloromethane : raethylalcohol - 96:4), followed by HPLC to give the title compounds $ (0.18 g) and 4 (0.08 g). Molecular wefghhof the compound was determined by FAB MS (ZMS AX 505H) analysis. To confirm the molecular structure, Nuclear Magnetic Resonance (NMR) measurements were performed on 600 MHz (Bruker) for 'H-NMR and on 150 MHz (Bruker) for UC-NMR» and the spectra are shown in Figs. 7 and 8, respectively. 1-5:Synthesis.of jD-2-(meuvltfaio)-Sari1 cvclosporin A: Compound 5 According to the general method, aLkylation was performed employing THF (100 ml), (i-Pr^NH (1.6 ml), BuLi (4.0 ml), cyclosporin A (1.0 g) in 30 ml THF and methyl disulfide 0^282) (1 -5 nil). The solution was stirred for 14 hrs at 0 °C and added with 20 ml water, followed by concentration. The residue was added with ether (Et-0), washed with water and a solution of saturated sodium chloride in sequence, and dried over anhydrous MgSCU. .Alter concentrating, the residue was subjected to silica gel column chromatography (100 g silica gel, dichloromethane : methylalcohol = 50 : 1 ~ 96 : 4), followed by HPLC to give the title compounds 5 (0.36 g) and 6 (0.05 g). Molecular weight of the compound was determined by FAB MS (ZMS AX 505H) analysis. To confirm the molecular structure, Nuclear Magnetic Resonance (NMR) measurements were performed on 600 MHz (Bruker) for 'H-NMR and on 150 MHz (Bruker) for 13C-NMR, and the spectra are shown in Figs. 9 and 10, respectively. 1-6: Synthesis of fN-methvl-Q-propenvl-D-Ser3] cvclosporin A: Compound 6 According to the general method, p-methylserine3] cyclosporin A (0.62 g, 0.5 mmol), tetrabutylammonium chloride (0.11 g, 0.5 mmol), and aryl bromide (0.24 g, 2.0 mmol) were dissolved in dichloromethane (50 ml), then added with 30 % NaOH (1.5 ml), and the mixture was stirred for 2 hrs. After adding with 50 ml dichloromethane, the solution was washed with water and a solution of saturated sodium chloride in sequence, and dried over anhydrous MgSO.». The concentrated residue was subjected to silica gel column chromatography (100 g silica gel, dichloromethane : methylalcohol = 97 : 3), followed by HPLC to give 0.4 g of the title compound. Molecular weight of the compound was determined by FAB MS (ZMS AX 505H) analysis. To confirm the molecular structure, Nuclear Magnetic Resonance (NMR) measurements were performed on 600 MHz (Bruker) for 'H-NMR and on 150 MHz (Bruker) for 13C-NMR, and the spectra are shown in Figs. 11 and Irrespectively. ]-7: Synthesis of fN-methvl-D-Ser3'! cvclosporin A: Compound 7 According to the general method, to a solution of 10 equivalents of LDA was added 1.0 g cvclosporin A far50 ml THF at -78 °C. The reaction mixture was stirred for 2 hrs at -78 °C and added with 10 g parafbrmaldehyde. After the temperature of the solution reached room temperature, the solution vras further stirred tor 24 hrs and added with 20 ml water, followed by concentration. The residue was addedwith ether (EtzO), washed with water and a solution of saturate! sodium chloride in sequence, and dried over anhydrous MgS04. After concentrating, the residue was subjected to silica gel column chromatography (100 g silica gel, dichloromethane : rnethylalcohol = 96 : 4), followed by HPLC to give 0.3 g of the title compound Molecular weight of the compound was determined by FAB MS (ZMS AX 505H) analysis. To confirm the molecular structure, Nuclear Magnetic Resonance (NMR) measurements were performed on 600 MHz (Bruker) for 'H-NMR and on 150 MHz (Broker) for UC-NMR, and the spectra are shown in Figs. 13 and 14, respectively. Preparative example 1: hair tonic 1-1: Preparation of hair tonic containing rN-methvl-p-Abu'] cyclosporin A Individual ingredients were mixed and stirred, and the mixtures were completely dissolved to prepare three hair growth promoting tonics, with compositions as shown in Table 1 below. It was found that the composition 1 of Table 1 has a hair growth promoting effect at a level similar to a conventional hair tonic containing 0.1 % cyclosporin A, as evaluated in an animal (Figure Remove)experiment according to the Test Example described later. (unit weight %) Ingredients Comp. 1 Comp. 2 Comp. 3 1 –2: preparation offo’f tonic containing [N-rnethvl-D-Nv^ cyclosporin A Individual ingredients were mixed and stirred, and the mixtures were completely dissolved to prepare tote hair growth promoting tonics, with compositions as shown in Table 2bdow. It was found that the composition 1 ofTable2hasahairgrcwthpronx>tingcffcctat a level similar to a conventional hair tonic containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later. Table 2: Formulation of hair tonic (unit weight %) (Table Remove) cycjospQiinA Individual ingredients were mixed and stirred, and the mixtures were completely dissolved to prepare three hair growth promoting tonics, with compositions as shown in Table 3 below. It was found that the composition 1 of Table 3 has a hair growth promoting effect at a level similar to a conventional hair tonic containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later. Table 3: Formulation of hair tonic (Table Remove) 1-4: Preparation of hair tonic containing fD-2^mdhvianiino'toart-4-vnovt' yyclogporinA Individual ingredients were mixed and stirred, and the mixtures were completely dissolved to prepare three hair grov-th promoting tonics, with compositions as shown in Table 4 below. It was found that the composition 1 of Table 4 has a hair growth promoting effect at a level similar to a conventional hair tonic' containing C.I % cyclosporin A, as evaluated in an animal experiment according to the Tes: Example described later. Table 4: Formulation of hair tonic (Table Remove) Individual ingredients were mixed and stirred, and the mixtures were completely dissolved to prepare three hair growth promoting tonics, with compositions as shown in Table 5 below. It was found that the composition 1 of Table 5 has a hair growth promoting effect at a level similar to a conventional hair tonic containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later. Table 5: Formulation of hair tonic (unit weight %) (Table Remove) (Table Remove) 1-6: Preparation of hair tonic containing rK-methvl-O-propenvl-D-Serl cvclosoorin A Indi\idual ingredients were mixed and stirred,' and the mixtures were completely dissolved to prepare three hair growth promoting tonics, with compositions as shown in Table 6 below. It was found that the composition 1 of Table 6 has a hair growth promoting effect at a level similar to a conventional hair tonic containing 0.1 % cyclosporin A. as evaluated in an animal experiment according to the Test Example described later. Table 6: Formulation of hair tonic (Table Remove) 1-7: Preparation of hair tonic containing IN-methvl-D-Ser3! cvclosporin A Individual ingredients were mixed and stirred, and the mixtures were completely dissolved to prepare three hair growth promoting tonics, with compositions as shown in Table 7 below. It was found that the composition 1 of Table 7 has a hair growth promoting effect at a level similar to a conventional hair tonic containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later. Table 7: Formulation of hair tonic (unit: weight '/•> \(Table Remove) Preparative example 2: hair cream 2-1: Preparation of hair cream containing [N-methvl-D-Abu3! cyclosporin A Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature, Additives such as flavor and colorant were admixed to prepare three hair creams, with compositions as shown in Table 8 below, Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients. It was found that the composition 1 of Table 8 has a hair growth promoting effect at a level similar to a conventional hair cream containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later. Table 8: Formulation of hair cream (unit weight %) (Table Remove) [N-nethyl-D-Abu^tydospotir.A 0.1 !.0 8.0 glycerin 7.0 ".0 7.0 dipropyleneglycol 20.0 20.C 20.0 polysfiyl«neg!ycol 5.0 5.0 5.0 wa~r balance not including flavor ar.d colc,-3rt; flavcr rvpical typical typical coicrant typical typical typical 2-2: Preparation of hair cream containing [N-methvl-D-Nva31 cvclosporin A Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hair creams, with compositions as shown in Table 9 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients. It was found that the composition 1 of Table 9 has a hair growth promoting effect at a level similar to a conventional hair cream containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later. Table 9: Formulation of hair cream (unit weight %) (Table Remove) 2-3: .Preparation. of hair cream containing fD-2-fmethvlarnmo/aexa4'V'novl5] cyclosporin A Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating :o 80 °C. Twc phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hair creams, with compositDns as shown in Table 10 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients. It was found that the composition 1 of Table 10 has a hair growth promoting effect at a level similar to a conventional hair cream containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later Table 10: Formulation of hair cream (unit: weight %) (Table Remove) 2-4: Preparation of hair cream containing [P.2-(methvlarnino')pent-4-vnovl>1 ri> gyclosporinA Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as fiavor and colorant were admixed to prepare three hair creams, with -compositions as shown in Table 11 below. Water was added to adjus: to ICO % the total weight including the oil-phase and water-phase ingredients. !: was found that the composition 1 of Table 11 has a hair growth promoting effect at a level similar to a conventional hair cream containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later. Table 11: Formulation of hair cream (Table Remove) 2-5: Preparation of hair cream containing D-l-methvlthio-Sar3! cvclosporin A Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hair creams, with compositions as shown in Table 12 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients. vf It was found that the composition 1 of Table 12 has a hair growth promoting effect at a level similar to a conventional hair cream containing 0.1 % cyclosporin A, as evaluated in an n-iwirrv Table 12: Formulation of hair cresm (unit weigh::, •':) Ingredients Cera. 1 Comp. 2 Corrg. 3 paraffin 5.0 5.0 3.0 csastearyl alcohol 5.5 5.5 5.5 perolaam 5-5 5.5 5.5 giycerin monostearaie 3.0 3.0 3.0 polyoxyethyleneocty'ldodecylether 3.0 3.0 3.0 propylparaben 03 0.3 0.3 ^-metfayltiiio-Sar^cyclosporinA 0.1 1.0 8.0 glycerin 7.0 7.0 7.0 dipropyleneglycol 20.0 20.0 20.0 polyttfaylenegiycol 5.0 5.0 5.0 wattr balance no: including flavor and colorant flavor tyPical ^P1^ W^ colorant typical typical typical 2-6: Preparation of hair cream containing rN-methvl-O-sropenvl-D-Ser^ cyclosporin A Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such, as flavor and colorant were admixed to prepare three hair creams, with compositions as shown in Table 13 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients. It was found that the composition 1 of Table 13 has a hair growth promoting effect at a level similar to a conventional hair cream containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later. Table 13: Formulation ofhair cream (unicw=ght%) (Table Remove) giyeerir. monostearate pciyox>-;iyleneoctyldodsc>'.rr.er prtjpylparaben [N-metyI-0-propenyI-D-Ser Jj cydospcrir. A glycerir. dpropy'.eneglycol pcfyeiyleneglycol 3.0 3.0 13 J.I -.0 20.0 5.0 5.0 3.0 0.3 3.0 7.0 20.G 5.0 £avor cclorar.t typical typical typicc. tvpici balance net including flavor and colorant typical typical 2-7: Preparation of hair cream containing fN-rrigLhyl-D-Ser3] cyclosporin A Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsif ed, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hair cream, with compositions as shown in Table 14 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients. It was found that the composition 1 of Table 14 has a hair growth promoting effect at a level similar to a conventional hair cream containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later. Table 14: Formulation of hair cream (unit weight %) (Table Remove) Table 16: Formulation of sharr.poc ('-.;: we: Hht%) (Table Remove) cvclosporinA All individual ingredients, except flavor, colorant and water, were mixed and the mixture was completely dissolved by heating, while stirring. After cooling to room temperature, the mixture was mixed with flavor and colorant. Water was finally added to adjust to 100 % the total weight, to prepare three shampoos, with compositions as shown in Table 17 below. Table 17: Formulation of shampoo ^ (unit weight %) Ingredients Comp. 1 Comp. 2 Comp. 3 sodium POE lauiyisuliuric add 40.0 40.0 40.0 (30 wt% aqueous solution) * palmci&CyaciddJEtharjplamide 3.0 3.0 3.0 Propyfenegtycol * 10 10 10 02 02 02 Btbno) 10 10 10 [D-2Kme&yfarnb»)hex»4.^^ 10 3.0 10.0 •fcyticnd 03 03 QJ (Table Remove) 3-4: Preparation of shampoo containing rD-2-dneLhvlamir.o'n3ent-4-\-novl3] cyclosporinA All individual ingredients, except flavor, colorant and water, were mixed and the mixture was completely dissolved by heating, while stirring. After cooling to room temperature, the mixture was mixed with flavor and colorant Water was finally added to adjust to 100 % the total weight, to prepare three shampoos, with compositions as shown in Table 18 below. Table 18: Formulation of shampoo (unit weight %) (Table Remove) All individual ingredients, except Havoc, colorant and water, were mixed and the mixture was completely dissolved by heating, while stirring. After cooling to room temperature, the mixture was mixed with flavor and colorant \Vatcr was finally added to adjust to 100 % the total weight, to prepare three shampoos, with compositions as shown in Table 19 below. Table 19: Formulation of shampoo (unit: weight %) COCT. 1 Comp.2 Comp. 3 (Table Remove) 3-6: Preparation of shampoo containing rN-rnethvl-0-propenvl-D-Ser] cyclosporin A All individual ingredients, except flavor, colorant and water, were mixed and the mixture was completely dissolved by heating, while stirring. After cooling to room temperature, the mixture was mixed with flavor and colorant Water was finally added to adjust to 100 % the total weight, to prepare three shampoos, with compositions as shown in Table 20 below. Table 20: Formulation of shampoo (unit weight %) (Table Remove) (Table Remove) 3-7: Preparation of shampoo containing [N-methvl-D-Ser3] cvclospprin A All individual ingredients, except flavor, colorant and water, were mixed and the mixture was completely dissolved by heating, while stirring. Aiier cooling to room temperature, the mixture was mixed with flavor and colorant. Water was finally added to adjust to 100 % the total weight, to prepare three shampoos, with compositions as shown in Table 21 below. Table 21: Formulation of shampoo (unit: weight %) (Table Remove) Individual ofl^phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by beating to 80 °C Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as Savor and colorant were admixed to prepare three hair conditioners, with compositions as shown in Table 22 below. Water was added to adjust to 100% the total weight including the oil-phase and water-phase ingredient. Table 22: Formulation of hair conditioner (unit we.ght %) (Table Remove) stearyldimethyl benzylammonium chloride (25 wt% aqueous solution) methyl paraoxybenzoic add salicylic add L-menthol water flavor colorant 8.0 0.2 03 0.3 balance typical typical 8.0 02 0.3 0.3 balance typical typical 8.0 0.2 0.3 0.3 balance typical typical 4-2: Preparation of hair conditioner containing [N-methvl-D-Nva"1] cyclosporin A Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hair conditioners, with compositions as shown in Table 23 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients. Table 23: Formulation of hair conditioner (unit weight %) (Table Remove) steayldiiEetbyl benzylammorjum chicrid; (25 wt°/: aqueous soiutior.) 8.0 S.O (Table Remove) cvclosporinA Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hak conditioners, \\ith compositions as shown in Table 24 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients. Table 24: Formulation of hair conditioner (Table Remove) 4-4: Preparation of hair conditioner containing fD-2-fmethvlarriino)peat-4-vnovl3' cvclosporin Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives :such as flavor and colorant were admixed to prepare three hair conditioners, with compositions as shown in Table 25 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients. Table 25: Formulation of hair conditioner (Table Remove) Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant wop admixed to prepare three hair conditioners, with compositions as shown In Table 26 below. Water was added to adjust to 100 % the total weight including the ofl-phasc and water-phase ingredients. Table 26: Formulation of hair conditioner (Table Remove) Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hair conditioners, with compositions as shown in Table 27 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients. Table 27: Formulation of hair conditioner (Table Remove) (25 wt% aqueous solution) (Table Remove) Individual oil-phase and \vater-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hair conditioners, with compositions as shown in Table 28 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients. Table 28: Formulation of hair conditioner (unit weight %) (Table Remove) the invention Female C57BL/6 mice of ages 6 to 7 weeks were utilized. After removing hairs or. the middle of the back with an electric shaver, the mice were weighed and randomly assigned to the test groups with an even distribution of weights. The mice were given one day for adaptation. From the next day, mice were applied once a day on their backs with cyclosporir. A and the cyclosporin A derivatives (Compounds 1 to 7) prepared by HPLC in Example 1 in amounts of 100 ul (cone. 0.1% w/v) for 30 days. "Hie results were determined by visual approach, in terms of degrees of hair regrowth. With respect to respective hair-removed .areas, rates of new hair growth were examined and compared. As can be seen in Table 29, cyclosporin derivatives of the Invention have a significant hair growth promoting effect, compared to the control in which mice were applied with a vehicle only. Further, the derivatives show a similar level of hair growth promoting effect, with respect to cyclosporin A Meanwhile, over a course of 30 days, as comparing the appearance of the backs, the mice of the control and all test groups showed no specific skin irritation. Table 29: Evaluation of cyclosporin derivatives based on hair regrowth in mice Compound vehicle cyclosporin A 1234567 applied Area rate ofhair 35 91 95 91 95 96 93 94 90 regrwih(%) On the basis of the foregoing results, the cyclosporin derivatives of the invention may be formulated in any form including liquid formulations, sprays, gels, pastes, emulsions, creams, conditioners, shampoos, and the like. A variety of forms are available though, considering their high commercial demand, hair tonics, creams, conditioners, and shampoos are provided herein. As revealed in the above the Test Example, the cyclosporin derivatives exhibit an excellent hair growth promoting effect, compared to the control Industrial Applicability ** . As apparent from the above description, the present invention provides a hair growth promoting agent comprising a cyclosporin A derivative substituted in the 3-position of cyclosporin A as an active ingredient, which exhibits an excellent hair growth promoting effect Claims 1. A hair growth promoting agent comprising a 3-posnon analog of cyclosporin represented by Formula 1 , as an active ingredient [Formula 1] A-B-C-D-E-F-G-H-I-J-K in which A represents N-methyl-(4R)-4-[(E>2-butenyl]-4-methyl-L-threonine, (2S,3R,4R,6E)-3-sulihydryl^rnethyl-2-(rnethylarnino)-6-octenoic acid or (2S,4R,6E)-3-oxo4-methyl-2-(methylamino)-6octenoic acid; B represents L-aminobutyric acid (Abu), L-alanine (Ala), L-threonine (Thr), L-valine (Val) or L-norvaline (Nva); C represents a D-amino acid represented by the general formula 1 , [General formula 1] CH3NH-CH(R>COOH in which, R is one selected from the group consisting of hydrogen, Q-Ce straight or branched alkyl, alkenyl or alkynyl moieties, substituted or unsubstituted with one or more selected from the group consisting of amino, hydroxy, halo, haloalkyl, ester, alkoxy, cyano, nitro, alkylamino, and dialkyiamino, and -X- R1 represented by the general formula 2 below, [General formula 2] -X-R1 in which, X is oxygen or sulfur, and R1 is one selected from the group consisting of hydrogen, and Ct-Q straight or branched alkyl, alkenyl or alkynyl moieties, substituted or unsubstituted with one or more selected from the group consisting of amino, hydroxy, halo, haloalkyl, ester, alkoxy, cyano, * nitro, alkylamino, and dialkylamino; D represent N-metfayl-L-leucine, y-hydroxy N-mcthyl-L-leucine or L-valine; E represents L-valine orL-oorvaline; F represents N-methyi-L-leucine, -y-irydroxy N-methyl-L-leucine or L-leucine; G represents L-alanine or L-alanine thkamide d V CS-NH], NH-CHCHj-CS-); H represents a D-aniino acid represented by the general formula 3. [General formula 3] -NH-CH(CH2R)-COOH in which, R is hydrogen or -X- R'represented by the general formula 4, [General formula 4] -X-R' in which, X is oxygen or sulfur, and R' is one selected from the group consisting of hydrogen, and Cj-Q straight or branched alkyl, alkenyl or alkynyl moieties, substituted or unsubstituted with one or more selected from the group consisting of amino, hydroxy, halo, haloalkyl, ester, alkoxy, cyano, nitro, alkylamino, and dialkylamino; I represents N-methyl-L-leucine, y-hydroxy-N-methyl-L-leucine or L-leucine; J represents N-methyl-L-leucine, -/-hydroxy-N-methyl-L-leucine or L-leucine; and, K represents N-methyl-L-valine or L-valine. 2. The hair growth promoting agent as set forth in claim 1, wherein the 3-position analog of cyclosporin is represented by Formula 2: [Formula 2] MeBmt-A'-e'-C'-O'-E'-F'-G'-H'H'-MeVal in which MeBmt represents N-methyl-(4R)-4-[(E)-2-butenyl]-4-rnethyl-L-tnreonijie; A1 represents L-aminobutyric acid, L-alanine, L-threonine, L-valine or L-norvaline; B' represents N-methyl-I>arrur»butyric add, N-methyl-D-norvaline, I>-2-(mdhylamino)hexa4-ynoyl, D-2-2-methylthio-sarcosine, N-methyl-OpropenyH>«rine or N-mefliyl-D-serine; Crqpresents N-methyl-L-lcucine, y^ydroxy-N-methyl-L-leiicine or L-valine; DrcprcsoilsUvalinecrlxxxvaline; E lepieseuts NnnShyl-L-leucine, fliydroxy N-mdhyl-L-kaicine or L-lcucine; P represerts L-alanine or L-alanine lirioamide dV CS-NH], s D-aianine cr D-serine; H1 represents N-methyl-L-leucine, y-hydroxy N-rnethyl-L-leucine cr L-!eucine; f represents N-methyl-L-leucine, y-hydroxy N-rnethyl-L-leucine or L-leucine; and. MeVal represents N-methyl-L-vaiine. 3. The hair growth promoting agent as set forth in claim 1, wherein the 3-position analog of cyclosporin is represented by Formula 3: [Formula 3] MeBmt-A'-B'-MeLetj-Val-MeLeu-Ala-DAIa-MeLeu-MeLeu-MeVe: I \| in which MeBmt represents N-methyl-(4R>4-[(E)-2-butenyl]-4-methyl-L-threonine; A" represents L-alanine, L-threonine, L-valine or L-norvaline; B" represents N-methyl-D-aniinobutyric acid, N-methyl-D-norvaline, D-2-(methylamino)hexa-4-ynoyl, D-2^methylamino)pent-4-ynoyl, D-2-methylthio-sarcosine, N- methyl-O-propenyl-D-serine or N-methyl-D-serine; MeLeu represents N-methyl-L-leucine; Val represents L-valine; Ala represents L-alanine; DAla represents D-alanine; and, MeVal represents N-methyl-L-valine. 4. The hair growth promoting agent as set forth in claim 1, comprising [N-methyl-D- Abu3] cyclosporin A as an active ingredient 5. The hair growth promoting agent as set forth in claim 1, comprising [N-methyl-D- Nva3] cyclosporin A as an active ingredient * w «. 6. The hair growth promoting agent as set forth in claim 1, comprising p-2- (methylamino)bex»4-ynoylj] cyclosporin A as an active ingredient 7. The hair growth promoting agent as set forth in claim 1, comprising [D-2- (methylarnino)pent-4-ynoyl3j cyclosporin A as an active ingredient. 8. The hair growth promQting ager. as set forth in claim 1, comprising [D-2-methylthio-Sar3] cyclosporin A as an active ingredient. 9. The hair gro\vth promoting agent as set fonh in claim 1, comprising [N-rr.e±yl-0- propenyl-D-Ser3] cyclosporin A as an active ingredient. 10. The hair growth promoting agent as set forth in claim 1. comprising [N-methyl- D-Ser3] cyclosporin A as an active ingredient 11. The hair growth promoting agent as set forth in any one of claims 1 to 10, which is formulated in a form selected from the group consisting of liquid formulation, spray, gel, paste, emulsion, cream, conditioner and shampoo. A hair growth promoting agent comprising a 3-position analog of cyclosporin represented by formula 1, substantially as herein described with reference to the foregoing description and drawings.

Full Text

USE OF 3-POSITION CYCLOSPORIN DERTVATI\"ES FOR HAIR GROWTH
Technical Field
The present invention relates to a hair growth promoting agent comprising a cyclosporin derivative as an active ingredient and more particularly, to a hair growth promoting agent comprising cyclosporin derivatives modified in the 3-position as an active ingredient.
Background Art
On average, the human scalp contains about 100,000 to 150,000 hairs. Each hair has three main stages of growth: anagen, catagen and telogen, after which the hair falls out. This hair growth cycle is repetitive and the duration of one cycle is different from other cycles, ranging approximately 3 to 6 years. Thus, the average adult normally loses about 50 to 100 hairs every day. In general, alopecia refers to a phenomenon wherein duration of the anagen growth phase is shortened and the percentage of hairs in the catagen and telogen phases increases, whereby the number of lost hairs is increased excessively and abnormally.
There are many theories to explain for loss of hair, including for example, poor blood circulation, excessive functioning of male sex hormone, excessive production and secretion of sebum, deterioration of scalp by peroxides, bacteria, etc., hereditary factors, aging, stress, etc. However, explicit mechanisms have not been revealed. Recently, the population suffering from hair loss is tending to increase, since changing dietary habits and stress imposed on individuals due to modern social environments, etc. has increased. Also, the age of the individuals affected by alopecia is dropping and furthermore, the population of female alopecia sufferers is rising.
One of preparations which are most commonly used for treatment and
M*
prevention of alopecia*is one that contains minoxidil. There are two hair-regrowth agents which have received approval from the U.S. Food and Drug Administration, and minoxidil is one of those approved hair-regrowth agents. Minoxidil was originally developed as a hypertension drug for the purpose of reducing blood pressure. However, when using this drug, as a side effect, a trichogenous effect

was observed and thereafter, this drug became famous as a hair-regrowth agent. Although mechanisms by which minoxidil works as a hr.r-regrowth asent is no: clearly understood, it is inferred that minoxidil increases blood flow by expansion of blood vessels, whereby roots of hairs are supplied with more nutrition and eventually, growth of hairs are promoted,
Such a model of bicod flow increase has been indirectly supported by a recent report that minoxidil enhances the expression of vascular endothelial growth factor (VEGF), a growth factor associated with vasodiiata:ion in the dermal papilla which is a main cell making up the hair roots. Also, other than the vasodilative effect of the minoxidil in the hair-restoring mechanism, it has been reported that minoxidil promotes activation of dermal papilla cells in the roots of hair incubated in vitro, and growth of hair follicles in a tissue culture of follicles in vitro. These facts indicate that minoxidii may work directly on the roots of hair as a growth factor.
In addition, finasteride, a main component of Propecia which has started to be sold by Merck, is used for treatment of alopecia. It inhibits conversion of the male hormone testosterone into dihydrotestosterone, which is a more potent male hormone than testosterone, On December of 1997, the 1 mg finasteride tablet was approved by the US FDA as a hair-regrowth agent for treatment of male pattern hair loss in men only, and is now commercially available. In clinical studies, it has been demonstrated to have a significant trichogenous effect. However, there has been a report that finasteride may inhibit male sexual function as a side effect. Since neither finasteride nor minoxidil show superior effect in clinical tests, and there is concern about side effects, many researches are conducted to develop a new and improved hair-regrowth agents.
The cyclosporin family of drugs has immunosuppressive activity. It is also effective to inhibit growth of virus, fungus, protozoan, etc. and has various physiological effects such as nephrotoxicity, hepatotoxicity, hypertension, enlargement of periodontium, trichogenous effect, and so on, as side effects. Cyclosporin A, a representative cyclosporin, is a cyclic peptide having the following Chemical Formula, "which comprises 11 amino acids, including several N-methyl araino acids and D-alanine at No. 8 residue.

[Structure Formula 1]
pMeBmt-Abu-Sar-MeLeu—Val-MeLeu-Aia-DA!a-MeLeu —MeLeu—
1 23 4 5,6 78 5 10 11
where MeBmt is N-raethyl-(4R)-4-"k(E)-2-biitenyrj-4-methyl-L--Jireonhe, Abu is L-a-aminobutyric acid, Sar is sarco sine, MeLeu is N-methyl-L-leucir.e,Val is L-valine, Ala is L-alanine, DAla is D-alanine, MeVal is N-raethyl-L-valine.
The amino acid form of cyclosporin A of the above Chemical Formula 1 is L-configuration, unless otherwise specified. The residue numbering of amino acids starts from MeBmt and proceeds clockwise, i.e. 1 for MeBmt and 11 for the last MeVal (N-methyl-L-valine) as shown in the Structure Formula 1. Nomenclature of '.•arious derivatives including cyclosporins A to Z, follows methods commonly used (Helv. Chim. Acta, 1987, 70:13-36). For example, if Abu in the 2-position of cyclosporin A is substituted with L-alanine, L-threonine, L-valine or L-norvaline, the derivatives thus prepared are named cyclosporin B, cyclosporin C, cyclosporin D or cyclosporin G, respectively. Further, when the amino acid residues of the cyclosporin derivatives differ from those of cydcsporin A, the derivatives are named by describing the substituent For example, if sarcosine, being the amino acid residue 3 of cyclosporin A, is substituted with N-methyl-D-Abu3 or N-methyl-D-Nva3, the derivatives thus prepared are named [N-methyl-D-Abu3] cyclosporin A or [N-methyl-D-Nva3] cyclosporin A, respectively. Meanwhile, a common method for abbreviating amino acids is employed, that is, N-methyl-L-leucine is abbreviated by MeLeu, N-methyl-L-isoleucine by MeUe, N-methyl-L-Valine by MeVal, N-methyl-L-alanine by MeAla, N-rr.ethyl-L-norvaline by MeNva, L-leucine by Leu, L-isoleucine by He, sarcosine by Sar, L-serine by Ser, L-valine, Val, L-alanine by Ala, D-alanine by DAla, L-aminobutviic acid by Abu, L-threonine by Thr, and L-norvaline by Nva. Further, as for a derivative of cyclosporin which is substituted with sulfur instead of a carbonyl oxygen at the amino acid residue 7, the name of the derivative may be cyclosporin 7-thioamide or [V CS-NH] cyclosporin, according to different references (Helv. Chint Acta. 74: 1953-1990,1991; J. Org, Chem. 58:673-677,1993; J. Org. Chem. 59:7249-7258,1994).
So far, possible development of cyclosporin as a hair-regrowth agent has been studied by many research grouris. Particularly, researches involving animal hair reg^>wth tests, human alopecia artata (J. Am Acad. DerrnaioL, 1990,22^42-250), human male pattern alopecia (J. Am, Acad, DennatoL, 1990,22^51-253 and Skm PhanracoL, 1994,7:101-104), and inhibition effect of hair toss by chenwtherapy in animal models (Am. J. PathoL, 1997,150:1433-1441) have been widely conducted In comparative exrwimenrs on mouse's back, h is shown that cyclosporin has a hair regrowrh effect about 100 times superior to minoxidn Based on such

findings, there have been attempts to utilize cyclosporin as a treatment for male pattern alopecia, and many applications for patents have been filed
For example, Japanese Patent Publication Kokai Nos. Sho 60-243008, Sho 62-19512. and Sho 62-19513 disclose use of cyclosporin derivatives as a hair regrowth agent Also, Europe Patent Publication No. 0414632B1 teaches a cyclosporin derivative modified ir. the 8-position, and PCT Publication No. 93/1 7039 teaches isocyclosporin. Moreover, United States Patent No. 5,807,820 and British Patent No. 2,218,334A disclose cyclosporins with excellent transdermal absorption, pursuant to the use of cyclosporins as hair restorers.
Disclosure of the Invention
Therefore, the present invention has been made in view of the above problems associated with side effects of cyclosporin A, and it is an object of the present invention to provide a hair growth promoting agent comprising a cyclosporin derivative as an active ingredienti which exerts an excellent hair growth-promotion ability.
In accordance with one aspect of the present invention, the above and other objects can be accomplished by the provision of a hair growth promoting agent comprising a 3-position analog of cyclosporin represented by the below Formula 1, as an active ingredient, which is prepared by synthesizing a variety of derivatives thereof and evaluating their hair growth promoting effects, with an aim of developing a novel agent for promoting hair growth,
[Formula 1]
A-B-C-D-E-F-G-H-I-J-K
wherein:
A represents N-me^l^RM-tCE^-butenylj-^methyl-L-threonine, (2S3R,4R,6E)-3-sulfhydryl4-mefeyl-2^methylarnino)-6-octenoic add or (2S,4R,6E)-3-oxo-4-methyl-2-(mrthylaraino>6-octenoic acid;
B represents l^aminobutyric add (Abu), L-alanine (Ala), L-threonine (Thr), L-valine (Val) or L-ootvaline (Sva);
C represents a D-amino add represented by the general formula 1,
[General formula 1]
in which,
R is one selected from the group consisting of hydrogen, Q-Q straight or branched

alkyl alkenyl or alkynyl moieties, subnituted or unsubstituted with one or more selectee from the group consisting of ammo, hydrox;., halo, haloaikyl, ester, alkoxy, cyano, ritro, alkylamino, and dialkylamino, and -X- R' represented by the general formula 2 below. [General formula 2]
-X-R1 in which, X is oxygen or sulfur, and
R1 is one selected from the group consisting of hydrogen, and C;-Cs straight or branched alkyl, alkenyl or alkynyl moieties, substituted or unsubsu'tuted with one or more selected from the group consisting of amino, hydroxy, halo, haloaikyl. ester, alkoxy, cyano,
nitro, alkylamino, and dialkylamino;
D represents N-methyl-L-leucine, y-hydroxy N-methyl-L-leucine or L-valine;
E represents L-valine or L-norvaline;
F represents N-methyl-L-leucine, y-hydroxy N-methyl-L-leucine or L-leucine;
G represents L-alanine or L-alanine thioamide ([V CS-NH], NH-CHCH3-CS-);
H represents a D-amino acid represented by the general formula 3,
[General formula 3]
-NH-CH(CH2R>COOH
in which,
R' is hydrogen or X-R represented by the general formula 4,
[General formula 4]
-X-R1
in which,
X is oxygen or sulfur, and
R' is one selected from the group consisting of hydrogen, and C'-Ce straight or branched alkyl, alkenyl or alkynyl moieties, substituted or unsubstituted with one or more selected from the group consisting of amino, hydroxy, halo, haloaikyl, ester, alkoxy, cyano, nitro, alkylamino, and dialkylamino;
I represents N-methyi-L-leucine, Y-hydroxy-N-mcthyl-L-leucinetor L-leucine;
J represents N-rnethyl-L-leucine, Y-hydroxy-N-rncthyl-L-leucine or L-leucine; and,
*.
K represents N-methyl-L-valine or L-valine,
In accordance with snotber aspect of the invention, that is provided a hair growth promoting agent comprising a 3-postion analog of cydosporin with an excellent hair growth promoting effect, represented by Formula 2 below, as an active ingredient.

[Formula 2] MeBmt-A'-e'-C'-O'-E'-^'-G'-H'-l-MeVal
wherein:
MeBmt represents N-meihyl^4R)^[(E)-2-butenyl]4-rnethyl-L-threonine;
A' represents L-arnip.obu:yric acid, L-alarJne, L-threonhe, L-valr.e or L-norvalir.e:
B1 represents N-meuyl-D-arriinobur.Tic acid, X-methyi-D-norvaiie, D-2-;methylarruno)hexa-4-ynoyl, D-2-(methylamino)pent-4-ynoyl, D-2-methylthio-sarcosine, N-:nethyl-O-propenyl-D-serine or N-methyi-D-serine;
C' represents N-metbyl-L-leucine, y-hydroxy-N-meth\i-L-!eucine orL-vaiine;
D' represents L-valine or L-norvaline;
E represents N-methyl-L4eucine, y-hydroxy N-methyl-L-leucine or L-leucine;
F represents L-alanine or L-aianine thioamide ([7\(/S CS-NH], NH-CHCH:,-CS-);
G' represents D-alanine or D-serine;
H1 represents N-methyl-L-leucine, y-hydroxy N-methyl-L-leucine or L-leucine;
I represents N-methyl-L-leucine, y-hydroxy N-methyl-L-leucine or L-leucine; and,
MeVal represents N-methyl-L-valine.
In accordance with another aspect of the invention, there is provided a hair growth promoting agent comprising a 3-position analog of cyclosporin with an excellent hair growth promoting effect, represented by Formula 3 below, as an active ingredient,
[Formula 3]
MeBmt-A"-8"-MeLeu-Val-MeLeu-Ala-DAIa-MeLeu-MeLeu-MeVa!
wherein:
4
MeBmt represents N-metyK4R>4{(E>2-butenyi}4-methyl-L-threonine; A" represents Iralanine, L-threonine, L-valine or L-norvaline; B" represents N-norvaline, D-2-(medoylaraino)hcxa-4-YQoyi, I>2^methYlamino)pcnt-4-ynoyl, D-2-methylthio-sarcosiDc, N-
MeLeu represents N-raethyi-L-leudnc;

Val represents L-valine; Ala represents L-alanine; . DAla represents D-alanine; and, Me Val represents N-methyl-L-valine.
In accordance with ye: another aspect of the present invention, there is provided a hair growth promoting agent, whose composition comprising a 3-position analog of cyclosporin may be formulated in the form of liquid formualtions, sprays, gels, pastes. emulsions, creams, conditioners or shampoos.
Brief Description of the Drawings
The above and other objects, features and other advantages of the present invention will be more clearly understood from the following detailed description taken in conjunction with the accompanying drawing, in which:
Fig. 1 is a !H-NMR spectrum of [N-methyl-D-Abu3] cyclosporin A;
Fig. 2 is a 13C-NMR spectrum of [N-methyl-D-Abu3] cyclosporin A;
Fig. 3 is a 1H-NMR spectrum of [N-methyl-D-Nva3] cyclosporin A;
Fig. 4 is a 13C-NMR spectrum of [N-methyl-D-Nva3] cyclosporin A;
Fig. 5 is a !H-NMR spectrum of [D-2-(methylamino)hexa4-ynoyl3] cyclosporin A;
Fig. 6 is a 13C-NMR spectrum of p-2-(methylamino)hexa-4-ynoylJ] cyclosporin A;
Fig. 7 is a 1H-NMR spectrum of p-2-(methylamino)pent-4-ynoyl'>] cyclosporin A; Fig. 8 is a °C-NMR spectrum of p-2-(methylamino)pent-4-ynoyl3] cyclosporin A;
Fig. 9 is a 'H-NMR spectrum of p^methylthioySar5] cyclosporin A;
Fig. 10 is a 13C-NMR spectrum of [D-2-(methylthio)-Sar5] cyclosporin A;
Fig. 11 is a 'H-NMR spectrum of [N-methyl-O-propenyl-D-Ser3] cyclosporin A;
Fig. 12 is a 13C-NMR spectrum of [N-methyl-O-propenyl-D-Ser5] cyclosporin A;
Fig. 13 is a 'H-NMR spectrum of [N-methyl-D-Ser3] cyclosporin A; and
Fig. 14 is a I3ONMR spectrum of [N-methyl-D-Scr3] cyclosporin A.
4
Best Mode far Carrying Out the Invention
Hereinafter, the present invention wffl be described in detail, in conjunction with various examples. These examples arc provided only for illustrative purposes, and the present invention ts not to be construed as being limited to those examples,.

With the aim of developing a novel agent with hair growth promoting effect, the present inventors chemically synthesized a variety of 3-position analogs of cyclosrorin, and hair growth promoting effects thereof were examined. Thus, the invention provides a hair growth promoting agent comprising a cyclosporin derivative as an active ingredient,
Example 1: Synthesis of 3-position analog of cvclosporin
A general method for the alkylation of cyclosporin A was as follows. Tetrahydrofuran (THF) was added with diisopropyl amine ((i-PrhNH) and added with a solution of n-butyl lithium (BuLi) in hexane under nitrogen atmosphere at -7S °C, followed by stirring for 30 mia To the solution of LDA (lithium diisopropylamide) thus prepared, cyclosporin A in THF was added, stirred for 1 hr, and electrophile was added.
1-1: Synthesis of fN-rneflivl-D-AbuJ] cvclosporin A: Compound 1 According to the general method above, to a solution of 10 equivalents of LDA was added 1.0 g cyclosporin A in 50 ml THF at -78 °C. The reaction mixture was stirred for 2 hrs at -78 °C and added with 0.4 ml ethyliodide. After the temperature of the solution reached room temperature, the solution was further stirred for 24 hrs and added with 20 ml water, followed by concentration. The residue was added with ether (EtiO), washed with water and a solution of saturated sodium chloride in sequence, and dried over anhydrous MgSO.;. After concentrating, the residue was subjected to silica gel column chromatography (100 g silica gel, dichloromethane : methylalcohol = 96 : 4), followed by HPLC to give 0.1 g of the title compound
Molecular weight of the compound was determined by FAB MS (ZMS AX 505H) analysis. To confirm the molecular structure, Nuclear Magnetic Resonance (NMR) measurements were performed on 600 MHz (Bruker) for 'H-NMR and on 150 MHz (Bruker) for 13C-NMR, and the spectra are shown in Figs. 1 and 2, respectively.
j-2: Synthesis of fN-methvl-D-Nva3! cvclosporin A: Compound 2
4
According to the general method, to a solution of 10 equivalents of LDA was added 1.0 g cyclosporin A in-50 ml THF at -78 °C. The reaction mixture was stirred for 2 hrs at -78 °C and added with 0.41 ml propylkxlide. After the temperature of the solution reached room temperature, the solution was further stirred for 24 hrs and added with 20 ml water, followed by ccocentratioa The residue was added with ether (EtjOX washed with water and a solution of saturated sodium chloride in sequence, and dried over-anhydrous MgS04. After

concentrating, the residue was subjected to silica gel column chromatography (: 00 g silica gel, dichloromethane : methylalcohol = 96 : 4), followed by HPLC to give 0.12 g of the title compound. Molecular weight of the compound was determined by FAB MS (ZMS AX 505H) analysis, To confirm the molecular structure, Nuclear Magnetic Resonance (NMR) measurements were performec on 600 MHz (Bruker) for 'H-NMR and on 150 :MHz (Bruker) for 13C-NMR, and the spectra are shown in Figs. 3 and 4, respectively.
1-3: Synthesis P-2-(methvlarnmo)hexfl-4-vnovl3'l cvclosporin A: Compound 3 According to the general method, to a solution of 10 equivalents of IDA was added 1.0 g cyclosporin A in 50 ml THF at -78 °C. The reaction mixture was stirred for 2 hrs at -78 °C and added with 0.73 ml l-bromo-2-butyne. After the temperature of the solution reached room temperature, the solution was further stirred for 24 hrs and added with 20 ml water, followed by concentration. The residue was added with ether (Et20), washed with water and a solution of saturated sodium chloride in sequence, and dried over anhydrous MgSO*. After concentrating, the residue was subjected to silica gel column chromatography (100 g silica gel, dichlororaethane: methylalcohol = 96:4), followed by HPLC to give 0.13 g of the title compound Molecular weight of the compound was determined by FAB MS (ZMS AX 505H) analysis. To confirm the molecular structure, Nuclear Magnetic Resonance (NMR) measurements were performed on 600 MHz (Bruker) for 1H-NMR and on 150 MHz (Bruker) for 13C-NMR, and the spectra are shown in Figs. 5 and 6, respectively.
1-4: Synthesis of p-24methylamino)pent-4-ynovlJl cvclosporin A: Compound 4 According to the general method, alkylation was performed employing THF (200 ml), (i-Pr):NH (3.2 ml), BuLi (8 ml), cyclosporin A (3.76 g) in 50 ml THF and propargyl bromide (3.57 g). After the temperature of the solution reached room temperature, the solution was further stirred for 24 hrs and added with 40 ml water, followed by concentration. The residue was added with ether (EtaO), washed with water and a solution of saturated sodium chloride in sequence, and dried over anhydrous MgSCu. After concentrating, the residue was subjected to silica gel column chromatography (100 g silica gel, dichloromethane : raethylalcohol - 96:4), followed by HPLC to give the title compounds $ (0.18 g) and 4 (0.08 g). Molecular wefghhof the compound was determined by FAB MS (ZMS AX 505H) analysis. To confirm the molecular structure, Nuclear Magnetic Resonance (NMR) measurements were performed on 600 MHz (Bruker) for 'H-NMR and on 150 MHz (Bruker) for UC-NMR» and the spectra are shown in Figs. 7 and 8, respectively.

1-5:Synthesis.of jD-2-(meuvltfaio)-Sari1 cvclosporin A: Compound 5 According to the general method, aLkylation was performed employing THF (100 ml), (i-Pr^NH (1.6 ml), BuLi (4.0 ml), cyclosporin A (1.0 g) in 30 ml THF and methyl disulfide 0^282) (1 -5 nil). The solution was stirred for 14 hrs at 0 °C and added with 20 ml water, followed by concentration. The residue was added with ether (Et-0), washed with water and a solution of saturated sodium chloride in sequence, and dried over anhydrous MgSCU. .Alter concentrating, the residue was subjected to silica gel column chromatography (100 g silica gel, dichloromethane : methylalcohol = 50 : 1 ~ 96 : 4), followed by HPLC to give the title compounds 5 (0.36 g) and 6 (0.05 g). Molecular weight of the compound was determined by FAB MS (ZMS AX 505H) analysis. To confirm the molecular structure, Nuclear Magnetic Resonance (NMR) measurements were performed on 600 MHz (Bruker) for 'H-NMR and on 150 MHz (Bruker) for 13C-NMR, and the spectra are shown in Figs. 9 and 10, respectively.
1-6: Synthesis of fN-methvl-Q-propenvl-D-Ser3] cvclosporin A: Compound 6 According to the general method, p-methylserine3] cyclosporin A (0.62 g, 0.5 mmol), tetrabutylammonium chloride (0.11 g, 0.5 mmol), and aryl bromide (0.24 g, 2.0 mmol) were dissolved in dichloromethane (50 ml), then added with 30 % NaOH (1.5 ml), and the mixture was stirred for 2 hrs. After adding with 50 ml dichloromethane, the solution was washed with water and a solution of saturated sodium chloride in sequence, and dried over anhydrous MgSO.». The concentrated residue was subjected to silica gel column chromatography (100 g silica gel, dichloromethane : methylalcohol = 97 : 3), followed by HPLC to give 0.4 g of the title compound. Molecular weight of the compound was determined by FAB MS (ZMS AX 505H) analysis. To confirm the molecular structure, Nuclear Magnetic Resonance (NMR) measurements were performed on 600 MHz (Bruker) for 'H-NMR and on 150 MHz (Bruker) for 13C-NMR, and the spectra are shown in Figs. 11 and Irrespectively.
]-7: Synthesis of fN-methvl-D-Ser3'! cvclosporin A: Compound 7 According to the general method, to a solution of 10 equivalents of LDA was added 1.0 g cvclosporin A far50 ml THF at -78 °C. The reaction mixture was stirred for 2 hrs at -78 °C and added with 10 g parafbrmaldehyde. After the temperature of the solution reached room temperature, the solution vras further stirred tor 24 hrs and added with 20 ml water, followed by concentration. The residue was addedwith ether (EtzO), washed with water and a solution of saturate! sodium chloride in sequence, and dried over anhydrous MgS04. After

concentrating, the residue was subjected to silica gel column chromatography (100 g silica gel, dichloromethane : rnethylalcohol = 96 : 4), followed by HPLC to give 0.3 g of the title compound Molecular weight of the compound was determined by FAB MS (ZMS AX 505H) analysis. To confirm the molecular structure, Nuclear Magnetic Resonance (NMR) measurements were performed on 600 MHz (Bruker) for 'H-NMR and on 150 MHz (Broker) for UC-NMR, and the spectra are shown in Figs. 13 and 14, respectively.
Preparative example 1: hair tonic
1-1: Preparation of hair tonic containing rN-methvl-p-Abu'] cyclosporin A Individual ingredients were mixed and stirred, and the mixtures were completely dissolved to prepare three hair growth promoting tonics, with compositions as shown in Table 1 below. It was found that the composition 1 of Table 1 has a hair growth promoting effect at a level similar to a conventional hair tonic containing 0.1 % cyclosporin A, as evaluated in an animal (Figure Remove)experiment according to the Test Example described later.
(unit weight %)
Ingredients Comp. 1 Comp. 2 Comp. 3
1 –2: preparation offo’f tonic containing [N-rnethvl-D-Nv^ cyclosporin A

Individual ingredients were mixed and stirred, and the mixtures were completely dissolved to prepare tote hair growth promoting tonics, with compositions as shown in Table 2bdow. It was found that the composition 1 ofTable2hasahairgrcwthpronx>tingcffcctat a level similar to a conventional hair tonic containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later.

Table 2: Formulation of hair tonic
(unit weight %)
(Table Remove) cycjospQiinA
Individual ingredients were mixed and stirred, and the mixtures were completely dissolved to prepare three hair growth promoting tonics, with compositions as shown in Table 3 below. It was found that the composition 1 of Table 3 has a hair growth promoting effect at a level similar to a conventional hair tonic containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later.
Table 3: Formulation of hair tonic

(Table Remove) 1-4: Preparation of hair tonic containing fD-2^mdhvianiino'toart-4-vnovt'
yyclogporinA
Individual ingredients were mixed and stirred, and the mixtures were completely dissolved to prepare three hair grov-th promoting tonics, with compositions as shown in Table 4 below. It was found that the composition 1 of Table 4 has a hair growth promoting effect at a level similar to a conventional hair tonic' containing C.I % cyclosporin A, as evaluated in an animal experiment according to the Tes: Example described later.
Table 4: Formulation of hair tonic

(Table Remove) Individual ingredients were mixed and stirred, and the mixtures were completely dissolved to prepare three hair growth promoting tonics, with compositions as shown in Table 5 below. It was found that the composition 1 of Table 5 has a hair growth promoting effect at a level similar to a conventional hair tonic containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later.
Table 5: Formulation of hair tonic
(unit weight %)

(Table Remove)
(Table Remove) 1-6: Preparation of hair tonic containing rK-methvl-O-propenvl-D-Serl cvclosoorin
A
Indi\idual ingredients were mixed and stirred,' and the mixtures were completely dissolved to prepare three hair growth promoting tonics, with compositions as shown in Table 6 below. It was found that the composition 1 of Table 6 has a hair growth promoting effect at a level similar to a conventional hair tonic containing 0.1 % cyclosporin A. as evaluated in an animal experiment according to the Test Example described later.
Table 6: Formulation of hair tonic

(Table Remove) 1-7: Preparation of hair tonic containing IN-methvl-D-Ser3! cvclosporin A
Individual ingredients were mixed and stirred, and the mixtures were completely dissolved to prepare three hair growth promoting tonics, with compositions as shown in Table 7 below. It was found that the composition 1 of Table 7 has a hair growth promoting effect at a level similar to a conventional hair tonic containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later.
Table 7: Formulation of hair tonic
(unit: weight '/•>
\(Table Remove) Preparative example 2: hair cream
2-1: Preparation of hair cream containing [N-methvl-D-Abu3! cyclosporin A Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature, Additives such as flavor and colorant were admixed to prepare three hair creams, with compositions as shown in Table 8 below, Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients.
It was found that the composition 1 of Table 8 has a hair growth promoting effect at a level similar to a conventional hair cream containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later.
Table 8: Formulation of hair cream
(unit weight %)

(Table Remove)
[N-nethyl-D-Abu^tydospotir.A 0.1 !.0 8.0
glycerin 7.0 ".0 7.0
dipropyleneglycol 20.0 20.C 20.0
polysfiyl«neg!ycol 5.0 5.0 5.0
wa~r balance not including flavor ar.d colc,-3rt;
flavcr rvpical typical typical
coicrant typical typical typical
2-2: Preparation of hair cream containing [N-methvl-D-Nva31 cvclosporin A Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hair creams, with compositions as shown in Table 9 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients.
It was found that the composition 1 of Table 9 has a hair growth promoting effect at a level similar to a conventional hair cream containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later.
Table 9: Formulation of hair cream
(unit weight %)

(Table Remove)
2-3: .Preparation. of hair cream containing fD-2-fmethvlarnmo/aexa4'V'novl5] cyclosporin A
Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating :o 80 °C. Twc phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hair creams, with compositDns as shown in Table 10 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients.
It was found that the composition 1 of Table 10 has a hair growth promoting effect at a level similar to a conventional hair cream containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later
Table 10: Formulation of hair cream
(unit: weight %)

(Table Remove) 2-4: Preparation of hair cream containing [P.2-(methvlarnino')pent-4-vnovl>1
*ri>
gyclosporinA
Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as fiavor and colorant were admixed to prepare three hair creams, with -compositions as shown in

Table 11 below. Water was added to adjus: to ICO % the total weight including the oil-phase and water-phase ingredients.
!: was found that the composition 1 of Table 11 has a hair growth promoting effect at a level similar to a conventional hair cream containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later.
Table 11: Formulation of hair cream

(Table Remove) 2-5: Preparation of hair cream containing D-l-methvlthio-Sar3! cvclosporin A Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hair creams, with compositions as shown in Table 12 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients.
vf
It was found that the composition 1 of Table 12 has a hair growth promoting effect at a level similar to a conventional hair cream containing 0.1 % cyclosporin A, as evaluated in an
n-iwirrv Table 12: Formulation of hair cresm
(unit weigh::, •':)
Ingredients Cera. 1 Comp. 2 Corrg. 3
paraffin 5.0 5.0 3.0
csastearyl alcohol 5.5 5.5 5.5
perolaam 5-5 5.5 5.5
giycerin monostearaie 3.0 3.0 3.0
polyoxyethyleneocty'ldodecylether 3.0 3.0 3.0
propylparaben 03 0.3 0.3
^-metfayltiiio-Sar^cyclosporinA 0.1 1.0 8.0
glycerin 7.0 7.0 7.0
dipropyleneglycol 20.0 20.0 20.0
polyttfaylenegiycol 5.0 5.0 5.0
wattr balance no: including flavor and colorant
flavor tyPical ^P1^ W^
colorant typical typical typical
2-6: Preparation of hair cream containing rN-methvl-O-sropenvl-D-Ser^
cyclosporin A
Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such, as flavor and colorant were admixed to prepare three hair creams, with compositions as shown in Table 13 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients.
It was found that the composition 1 of Table 13 has a hair growth promoting effect at a level similar to a conventional hair cream containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later.
Table 13: Formulation ofhair cream
(unicw=ght%)
(Table Remove)
giyeerir. monostearate
pciyox>-;iyleneoctyldodsc>'.rr.er
prtjpylparaben
[N-metyI-0-propenyI-D-Ser Jj cydospcrir. A
glycerir.
dpropy'.eneglycol
pcfyeiyleneglycol

3.0
3.0
13
J.I
-.0
20.0
5.0

5.0 3.0 0.3 3.0 7.0 20.G 5.0

£avor cclorar.t

typical typical
typicc. tvpici
balance net including flavor and colorant
typical typical

2-7: Preparation of hair cream containing fN-rrigLhyl-D-Ser3] cyclosporin A Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsif ed, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hair cream, with compositions as shown in Table 14 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients.
It was found that the composition 1 of Table 14 has a hair growth promoting effect at a level similar to a conventional hair cream containing 0.1 % cyclosporin A, as evaluated in an animal experiment according to the Test Example described later.
Table 14: Formulation of hair cream
(unit weight %)
(Table Remove)
Table 16: Formulation of sharr.poc
('-.;: we: Hht%)
(Table Remove) cvclosporinA
All individual ingredients, except flavor, colorant and water, were mixed and the mixture was completely dissolved by heating, while stirring. After cooling to room temperature, the mixture was mixed with flavor and colorant. Water was finally added to adjust to 100 % the total weight, to prepare three shampoos, with compositions as shown in Table 17 below.
Table 17: Formulation of shampoo
^ (unit weight %)
Ingredients Comp. 1 Comp. 2 Comp. 3
sodium POE lauiyisuliuric add
40.0 40.0 40.0
(30 wt% aqueous solution) *
palmci&CyaciddJEtharjplamide 3.0 3.0 3.0
Propyfenegtycol * 10 10 10
02 02 02
Btbno) 10 10 10
[D-2Kme&yfarnb»)hex»4.^^ 10 3.0 10.0
•fcyticnd 03 03 QJ

(Table Remove) 3-4: Preparation of shampoo containing rD-2-dneLhvlamir.o'n3ent-4-\-novl3] cyclosporinA
All individual ingredients, except flavor, colorant and water, were mixed and the mixture was completely dissolved by heating, while stirring. After cooling to room temperature, the mixture was mixed with flavor and colorant Water was finally added to adjust to 100 % the total weight, to prepare three shampoos, with compositions as shown in Table 18 below.
Table 18: Formulation of shampoo
(unit weight %)

(Table Remove) All individual ingredients, except Havoc, colorant and water, were mixed and the mixture was completely dissolved by heating, while stirring. After cooling to room temperature, the mixture was mixed with flavor and colorant \Vatcr was finally added to adjust to 100 % the total weight, to prepare three shampoos, with compositions as shown in Table 19 below.
Table 19: Formulation of shampoo
(unit: weight %)
COCT. 1 Comp.2 Comp. 3
(Table Remove) 3-6: Preparation of shampoo containing rN-rnethvl-0-propenvl-D-Ser] cyclosporin A All individual ingredients, except flavor, colorant and water, were mixed and the mixture was completely dissolved by heating, while stirring. After cooling to room temperature, the mixture was mixed with flavor and colorant Water was finally added to adjust to 100 % the total weight, to prepare three shampoos, with compositions as shown in Table 20 below.
Table 20: Formulation of shampoo
(unit weight %)
(Table Remove)
(Table Remove) 3-7: Preparation of shampoo containing [N-methvl-D-Ser3] cvclospprin A All individual ingredients, except flavor, colorant and water, were mixed and the mixture was completely dissolved by heating, while stirring. Aiier cooling to room temperature, the mixture was mixed with flavor and colorant. Water was finally added to adjust to 100 % the total weight, to prepare three shampoos, with compositions as shown in Table 21 below.
Table 21: Formulation of shampoo
(unit: weight %)
(Table Remove) Individual ofl^phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by beating to 80 °C Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as Savor and colorant were admixed to prepare three hair conditioners, with compositions as shown in Table 22 below. Water was added to adjust to 100% the total weight including the
oil-phase and water-phase ingredient.
Table 22: Formulation of hair conditioner
(unit we.ght %)
(Table Remove)
stearyldimethyl benzylammonium chloride
(25 wt% aqueous solution) methyl paraoxybenzoic add salicylic add L-menthol water flavor colorant

8.0
0.2
03
0.3
balance typical typical

8.0
02
0.3
0.3
balance typical typical

8.0
0.2
0.3
0.3
balance typical typical

4-2: Preparation of hair conditioner containing [N-methvl-D-Nva"1] cyclosporin A Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hair conditioners, with compositions as shown in Table 23 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients.
Table 23: Formulation of hair conditioner
(unit weight %)
(Table Remove)
steayldiiEetbyl benzylammorjum chicrid; (25 wt°/: aqueous soiutior.)

8.0

S.O

(Table Remove) cvclosporinA
Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hak conditioners, \\ith compositions as shown in Table 24 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients.
Table 24: Formulation of hair conditioner

(Table Remove)
4-4: Preparation of hair conditioner containing fD-2-fmethvlarriino)peat-4-vnovl3' cvclosporin
Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives :such as flavor and colorant were admixed to prepare three hair conditioners, with compositions as shown in Table 25 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients.
Table 25: Formulation of hair conditioner

(Table Remove) Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant wop admixed to prepare three hair conditioners, with compositions as shown In Table 26 below. Water was added to adjust to 100 % the total weight including the ofl-phasc and water-phase ingredients.
Table 26: Formulation of hair conditioner

(Table Remove) Individual oil-phase and water-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hair conditioners, with compositions as shown in Table 27 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients.
Table 27: Formulation of hair conditioner

(Table Remove) (25 wt% aqueous solution)

(Table Remove) Individual oil-phase and \vater-phase ingredients were mixed in a separate container, and each mixture was completely dissolved by heating to 80 °C. Two phases of the ingredients were mixed, emulsified, and cooled to room temperature. Additives such as flavor and colorant were admixed to prepare three hair conditioners, with compositions as shown in Table 28 below. Water was added to adjust to 100 % the total weight including the oil-phase and water-phase ingredients.
Table 28: Formulation of hair conditioner
(unit weight %)
(Table Remove) the invention

Female C57BL/6 mice of ages 6 to 7 weeks were utilized. After removing hairs or. the middle of the back with an electric shaver, the mice were weighed and randomly assigned to the test groups with an even distribution of weights. The mice were given one day for adaptation. From the next day, mice were applied once a day on their backs with cyclosporir. A and the cyclosporin A derivatives (Compounds 1 to 7) prepared by HPLC in Example 1 in amounts of 100 ul (cone. 0.1% w/v) for 30 days. "Hie results were determined by visual approach, in terms of degrees of hair regrowth. With respect to respective hair-removed .areas, rates of new hair growth were examined and compared.
As can be seen in Table 29, cyclosporin derivatives of the Invention have a significant hair growth promoting effect, compared to the control in which mice were applied with a vehicle only. Further, the derivatives show a similar level of hair growth promoting effect, with respect to cyclosporin A Meanwhile, over a course of 30 days, as comparing the appearance of the backs, the mice of the control and all test groups showed no specific skin irritation.
Table 29: Evaluation of cyclosporin derivatives based on hair regrowth in mice
Compound
vehicle cyclosporin A 1234567
applied
Area rate ofhair
35 91 95 91 95 96 93 94 90
regrwih(%)
On the basis of the foregoing results, the cyclosporin derivatives of the invention may be formulated in any form including liquid formulations, sprays, gels, pastes, emulsions, creams, conditioners, shampoos, and the like. A variety of forms are available though, considering their high commercial demand, hair tonics, creams, conditioners, and shampoos are provided herein. As revealed in the above the Test Example, the cyclosporin derivatives exhibit an excellent hair growth promoting effect, compared to the control
Industrial Applicability
**
*.
As apparent from the above description, the present invention provides a hair growth promoting agent comprising a cyclosporin A derivative substituted in the 3-position of cyclosporin A as an active ingredient, which exhibits an excellent hair growth promoting effect

Claims
1. A hair growth promoting agent comprising a 3-posnon analog of cyclosporin represented by Formula 1 , as an active ingredient [Formula 1]
A-B-C-D-E-F-G-H-I-J-K
in which
A represents N-methyl-(4R)-4-[(E>2-butenyl]-4-methyl-L-threonine, (2S,3R,4R,6E)-3-sulihydryl^rnethyl-2-(rnethylarnino)-6-octenoic acid or (2S,4R,6E)-3-oxo4-methyl-2-(methylamino)-6octenoic acid;
B represents L-aminobutyric acid (Abu), L-alanine (Ala), L-threonine (Thr), L-valine (Val) or L-norvaline (Nva);
C represents a D-amino acid represented by the general formula 1 ,
[General formula 1]
CH3NH-CH(R>COOH
in which,
R is one selected from the group consisting of hydrogen, Q-Ce straight or branched alkyl, alkenyl or alkynyl moieties, substituted or unsubstituted with one or more selected from the group consisting of amino, hydroxy, halo, haloalkyl, ester, alkoxy, cyano, nitro, alkylamino,
and dialkyiamino, and -X- R1 represented by the general formula 2 below, [General formula 2] -X-R1 in which, X is oxygen or sulfur, and
R1 is one selected from the group consisting of hydrogen, and Ct-Q straight or branched alkyl, alkenyl or alkynyl moieties, substituted or unsubstituted with one or more selected from the group consisting of amino, hydroxy, halo, haloalkyl, ester, alkoxy, cyano,
**
nitro, alkylamino, and dialkylamino;
D represent N-metfayl-L-leucine, y-hydroxy N-mcthyl-L-leucine or L-valine; E represents L-valine orL-oorvaline;
F represents N-methyi-L-leucine, -y-irydroxy N-methyl-L-leucine or L-leucine; G represents L-alanine or L-alanine thkamide d V CS-NH], NH-CHCHj-CS-);

H represents a D-aniino acid represented by the general formula 3. [General formula 3] -NH-CH(CH2R)-COOH in which,
R is hydrogen or -X- R'represented by the general formula 4, [General formula 4]
-X-R'
in which,
X is oxygen or sulfur, and
R' is one selected from the group consisting of hydrogen, and Cj-Q straight or branched alkyl, alkenyl or alkynyl moieties, substituted or unsubstituted with one or more selected from the group consisting of amino, hydroxy, halo, haloalkyl, ester, alkoxy, cyano,
nitro, alkylamino, and dialkylamino;
I represents N-methyl-L-leucine, y-hydroxy-N-methyl-L-leucine or L-leucine; J represents N-methyl-L-leucine, -/-hydroxy-N-methyl-L-leucine or L-leucine; and, K represents N-methyl-L-valine or L-valine.
2. The hair growth promoting agent as set forth in claim 1, wherein the 3-position analog of cyclosporin is represented by Formula 2: [Formula 2]
MeBmt-A'-e'-C'-O'-E'-F'-G'-H'H'-MeVal
in which
MeBmt represents N-methyl-(4R)-4-[(E)-2-butenyl]-4-rnethyl-L-tnreonijie;
A1 represents L-aminobutyric acid, L-alanine, L-threonine, L-valine or L-norvaline;
B' represents N-methyl-I>arrur»butyric add, N-methyl-D-norvaline, I>-2-(mdhylamino)hexa4-ynoyl, D-2-2-methylthio-sarcosine, N-methyl-OpropenyH>«rine or N-mefliyl-D-serine;
Crqpresents N-methyl-L-lcucine, y^ydroxy-N-methyl-L-leiicine or L-valine;
DrcprcsoilsUvalinecrlxxxvaline;
E lepieseuts NnnShyl-L-leucine, fliydroxy N-mdhyl-L-kaicine or L-lcucine;
P represerts L-alanine or L-alanine lirioamide dV CS-NH], s D-aianine cr D-serine;
H1 represents N-methyl-L-leucine, y-hydroxy N-rnethyl-L-leucine cr L-!eucine; f represents N-methyl-L-leucine, y-hydroxy N-rnethyl-L-leucine or L-leucine; and. MeVal represents N-methyl-L-vaiine.
3. The hair growth promoting agent as set forth in claim 1, wherein the 3-position
analog of cyclosporin is represented by Formula 3:
[Formula 3]
MeBmt-A'-B'-MeLetj-Val-MeLeu-Ala-DAIa-MeLeu-MeLeu-MeVe:
I |
in which
MeBmt represents N-methyl-(4R>4-[(E)-2-butenyl]-4-methyl-L-threonine;
A" represents L-alanine, L-threonine, L-valine or L-norvaline; B" represents N-methyl-D-aniinobutyric acid, N-methyl-D-norvaline, D-2-(methylamino)hexa-4-ynoyl, D-2^methylamino)pent-4-ynoyl, D-2-methylthio-sarcosine, N-
methyl-O-propenyl-D-serine or N-methyl-D-serine; MeLeu represents N-methyl-L-leucine; Val represents L-valine; Ala represents L-alanine; DAla represents D-alanine; and, MeVal represents N-methyl-L-valine.
4. The hair growth promoting agent as set forth in claim 1, comprising [N-methyl-D-
Abu3] cyclosporin A as an active ingredient
5. The hair growth promoting agent as set forth in claim 1, comprising [N-methyl-D-
Nva3] cyclosporin A as an active ingredient *
w
«.
6. The hair growth promoting agent as set forth in claim 1, comprising p-2-
(methylamino)bex»4-ynoylj] cyclosporin A as an active ingredient
7. The hair growth promoting agent as set forth in claim 1, comprising [D-2-
(methylarnino)pent-4-ynoyl3j cyclosporin A as an active ingredient.
8. The hair growth promQting ager. as set forth in claim 1, comprising [D-2-methylthio-Sar3] cyclosporin A as an active ingredient.
9. The hair gro\vth promoting agent as set fonh in claim 1, comprising [N-rr.e±yl-0-
propenyl-D-Ser3] cyclosporin A as an active ingredient.
10. The hair growth promoting agent as set forth in claim 1. comprising [N-methyl-
D-Ser3] cyclosporin A as an active ingredient
11. The hair growth promoting agent as set forth in any one of claims 1 to 10, which
is formulated in a form selected from the group consisting of liquid formulation, spray, gel,
paste, emulsion, cream, conditioner and shampoo.
A hair growth promoting agent comprising a 3-position analog of cyclosporin represented by formula 1, substantially as herein described with reference to the foregoing description and drawings.

Documents:

01843-delnp-2003-abstract.pdf

01843-delnp-2003-assignments.pdf

01843-delnp-2003-claims.pdf

01843-delnp-2003-correspondence-others.pdf

01843-delnp-2003-drawings.pdf

01843-delnp-2003-form-1.pdf

01843-delnp-2003-form-18.pdf

01843-delnp-2003-form-2.pdf

01843-delnp-2003-form-3.pdf

01843-delnp-2003-form-5.pdf

01843-delnp-2003-pct-409.pdf

01843-delnp-2003-pct-request form.pdf

01843-delnp-2003-pct-search report.pdf

1843-DELNP-2003-Correspondence-Others-(05-03-2008).pdf

1843-DELNP-2003-Drawings-(05-03-2008).pdf

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Patent Number

222068

Indian Patent Application Number

01843/DELNP/2003

PG Journal Number

32/2008

Publication Date

08-Aug-2008

Grant Date

16-Jul-2008

Date of Filing

07-Nov-2003

Name of Patentee

LG HOUSEHOLD & HEALTH CARE LTD.

Applicant Address

20, YOUIDO-DONG, YOUNGDEUNGPO-GU, SEOUL 150-010, REPUBLIC OF KOREA.

Inventors:

#	Inventor's Name	Inventor's Address
1	KIM, SANG-NYUN	109-1005 SEJONG APT, 462-5 JEONMIN-DONG, YUSONG-GU, DAEJEON 305-728, REPUBLIC OF KOREA
2	AHN, HO-JEONG	107-1106 SEJONG APT, 462-5 JEONMIN-DONG, YUSONG-GU, DAEJEON 305-728, REPUBLIC OF KOREA
3	LEE, CHANG-WOO	106-404 SANGLOKSOO APT, MANNYON-DONG, SEO-GU, DAEJEON 302-150 REPUBLIC OF KOREA
4	LEE, MIN-HO	1-205 LG SATAEK, 386-4 DORYONG-DONG, YUSONG-GU, DAEJEON 305-340 REPUBLIC OF KOREA
5	KIM, JUNG-HUN	106-1006 HANVIT APT, EOEUN-DONG, YUSONG-GU, DAEJEON 305-755 REPUBLIC OF KOREA
6	KIM, JONG-IL	105-1302 SEJONG APT, 462-5 JEONMIN-DONG, YUSONG-GU, DAEJEON 305-728 REPUBLIC OF KOREA
7	KIM, SEUNG-JIN	270-276 GAEBONG 3-DONG, GURO-GU, SEOUL 152-093 REPUBLIC OF KOREA
8	CHO, HO-SONG	1-407 LG SATAEK, DORYONG-DONG, YUSONG-GU DAEJEON 305-340 REPUBLIC OF KOREA
9	LEE, HEON-SIK	1-405 LG SATAEK, DORYONG-DONG, YUSONG-GU, DAEJEON 305-340 REPUBLIC OF KOREA
10	KIM, HYUNG-JIN	1-108 LG SATAEK, DORYONG-DONG, YUSONG-GU, DAEJEON 305-340 REPUBLIC OF KOREA

PCT International Classification Number

A61K 7/06

PCT International Application Number

PCT/KR2002/00879

PCT International Filing date

2002-05-11

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1	2001/25682	2001-05-11	Republic of Korea