The present invention provides oligopeptides having morphogenesis promoting activity and in particular, hair promoting activity. The oligopeptides may be in monomer form, monomer having a reactive substance bound form or as a polymer, such as a dimer including a homodimer; heterodimer; homotrimer; or heterodimer. The present invention also provides monoclonal antibodies that specifically recognize a 220 kDa antigen of epithelial new hair follicles; hybridomas producing such antibody; and methods and kits for assaying hair growth in mammalian subjects.

TECHNICAL FIELD

The present invention relates to oligopeptides having morphogenesis activity. In particular, the present invention provides compositions comprising oligopeptides having hair growth promoting activity and methods for promoting hair growth in humans. The present invention also relates to monoclonal antibodies specific for an antigen of epithelial new hair follicle and methods for the evaluation of hair growth promoting activity using such a monoclonal antibody.

BACKGROUND

The normal morphogenesis of epithelial tissue has been suggested to be controlled by factors derived from mesenchymal cells present around the epithelial tissue. Diseases resulting from the abnormal morphogenesis of epithelial tissue are largely caused by abnormalities of mesenchymal cells. Therefore, an interest has arisen in understanding the mechanism by which mesenchymal cells control the morphogenesis of epithelial tissue.

Epimorphin, disclosed in Japanese Patent Laid-Open Publication No. 25295/94, has 277 to 289 amino acids as a core protein, and has the action of promoting the morphogenesis of epithelial tissue through its action on epithelial cells. It was found that normal tissue formation did not progress when epimorphin failed to function.

Epimorphin has been described in Hirai et al. (1992, Cell, 69:471 481); Hirai (1994, Eur. J. Biochem, vol. 225, 1133 1139); Hirai, et al. (1998, J. Cell. Biol., 140:159 169); and Hirai, et al. (2001, J. Cell. Biol., 153:785 794).

EP 0698666 A2 describes the structure of full length epimorphin as roughly divided into four fragments, beginning from the N-terminus, a coiled coil domain (1), a functional domain (2), a coiled coil domain (3), and hydrophobic domain at the C-terminal. EP0698666A discloses that the functional domain (the domain specified by 104th to 187th amino acids in human epimorphin) participates in cell adhesion and is associated with expression of physiological activity of epimorphin.

U.S. Pat. No. 5,726,298, issued Mar. 10, 1998, discloses human and murine epimorphin nucleotide and amino acid sequences. WO98/22505 and EP 1008603A1 describe polypeptides specified by the N-terminal sequence of the 1st to 103rd amino acids of human epimorphin and by the N-terminal sequence of the 1st to 104th amino acids of murine epimorphin.

Native mammalian epimorphin is almost insoluble in an aqueous media such as saline, which causes difficulty in using epimorphin in compositions for human treatment. Japanese Patent Laid-Open Publication No. 25295/1994 discloses a modified form of epimorphin obtained by removing a hydrophobic region at the C-terminus.

In spite of developments in the understanding of epimorphin and morphogenesis of epithelial tissue, there remains a need for means to modify the morphogenesis of epithelial tissue, in particular as it relates to diseases or disorders associated with abnormal morphogenesis.

DISCLOSURE OF THE INVENTION

The present invention relates to oligopeptides useful for the treatment or amelioration of symptoms of diseases or disorders associated with abnormal morphogenesis. The oligopeptides of the present invention can be used to induce morphogenesis, induce revascularization effect, induce regeneration effect, induce cardiovascular regeneration, and induce endothelial cell growth. The oligopeptides of the present invention can be used, for example, for the treatment of and/or amelioration of symptoms of burns or wounds or to promote hair growth or prevent hair loss. In particular, the present invention relates to oligopeptides having hair growth promoting activity and to methods of promoting hair growth. The present invention also relates to methods of assaying for hair growth using a monoclonal antibody specific for a 220 kDa antigen of epithelial new follicles and kits comprising a monoclonal antibody of the present invention.

The present invention provides isolated oligopeptides of between about 5 and about 104 amino acid residues in length having hair growth promoting activity, comprising the following amino acid sequence:

X1-X2-X3-X4-X5-X6-X7;

X1-X2-X3-X4-X6-X5-X7;

X1-X2-X3-X6-X4-X5-X7; or

X1-X2-X6-X3-X4-X5-X7;

wherein X1 is an amino acid residue of Ser, Ala, Tyr, Thr, Pro, Phe, Val, Gly, Leu, Ile or Met, or is deleted from said oligopeptide;

X2 is an amino acid residue of Ile, Gly, Asn, Thr, Val, Ser, Phe, Leu, Ala, Pro, Cys, or Met, or is deleted from said oligopeptide;

X3 is an amino acid residue of Glu, Lys, Gln, Arg, Ala, Val, Trp, Cys, or Asp;

X4 is an amino acid residue of Gln, Pro, Glu, Thr, Arg, Ser, His, Cys, or Lys;

X5 is an amino acid residue of Ser, Trp, Phe, Thr, Cys, Tyr, Pro, Ala, Gly, Val, Leu, Ile, or Met;

X6 is an amino acid residue Cys; a reactive substance-bound Cys or a reactive substance-bound Lys; and

X7 is an amino acid residue of Asp, Glu, His, Ser, Ala, Gly, Asn, Tyr, Arg, or Leu, or is deleted from said oligopeptide, with the proviso that the oligopeptide is not identical to SEQ ID NO:1 or SEQ ID NO:2.

The present invention also provides isolated oligopeptides of between about 5 and about 104 amino acid residues in length having hair growth promoting activity, comprising the following amino acid sequence,

X1-X2-X3-X4-X5-X6-X7;

X1-X2-X3-X4-X6-X5-X7;

X1-X2-X3-X6-X4-X5-X7; or

X1-X2-X6-X3-X4-X5-X7;

wherein X1 is an amino acid residue of Ser, Tyr, Thr, or Pro, or is deleted from said oligopeptide;

X2 is an amino acid residue of Ile, Asn, Thr, or Ser, or is deleted from said oligopeptide;

X3 is an amino acid residue of Glu, Ala, Trp, or Asp;

X4 is an amino acid residue of Gln;

X5 is an amino acid residue of Ser, Cys, or Tyr;

X6 is an amino acid residue of Cys; a reactive substance-bound Cys or a reactive substance-bound Lys; and

X7 is an amino acid residue of Asp, Ala, Gly, or Leu, or is deleted from said oligopeptide, with the proviso that the oligopeptide is not SEQ ID NO:2.

The present invention also provides isolated oligopeptides of between about 7 and about 100 amino acid residues in length having hair growth promoting activity, comprising the following amino acid sequence:

X1-X2-X3-X4-X5-X6-X7;

X1-X2-X3-X4-X6-X5-X7;

X1-X2-X3-X6-X4-X5-X7; or

X1-X2-X6-X3-X4-X5-X7;

wherein X1 is an amino acid residue of Ser, Ala, Tyr, Thr, Pro, Phe, Val, Gly, Leu, Ile or Met, or is deleted from said oligopeptide;

X2 is an amino acid residue of Ile, Gly, Asn, Thr, Val, Ser, Phe, Leu, Ala, Pro, Cys, or Met, or is deleted from said oligopeptide;

X3 is an amino acid residue of Glu, Lys, Gln, Arg, Ala, Val, Trp, Cys, or Asp;

X4 is an amino acid residue of Gln, Pro, Glu, Thr, Arg, Ser, His, Cys, or Lys;

X5 is an amino acid residue of Ser, Trp, Phe, Thr, Cys, Tyr, Pro, Ala, Gly, Val, Leu, Ile, or Met;

X6 is an amino acid residue Cys; a reactive substance-bound Cys or a reactive substance-bound Lys; and

X7 is an amino acid residue of Asp, Glu, His, Ser, Ala, Gly, Asn, Tyr, Arg, or Leu, or is deleted from said oligopeptide.

The present invention also provides isolated oligopeptides of between about 7 and about 100 amino acid residues in length having hair growth promoting activity comprising the following amino acid sequence:

X1-X2-X3-X4-X5-X6-X7;

X1-X2-X3-X4-X6-X5-X7;

X1-X2-X3-X6-X4-X5-X7; or

X1-X2-X6-X3-X4-X5-X7;

wherein X1 is an amino acid residue of Ser, Tyr, Thr, or Pro, or is deleted from said oligopeptide;

X2 is an amino acid residue of Ile, Asn, Thr, or Ser, or is deleted from said oligopeptide;

X3 is an amino acid residue of Glu, Ala, Trp, or Asp;

X4 is an amino acid residue of Gln;

X5 is an amino acid residue of Ser, Cys, or Tyr;

X6 is an amino acid residue of Cys; and

X7 is an amino acid residue of Asp, Ala, Gly, or Leu, or is deleted from said oligopeptide.

In some examples, an oligopeptide having hair growth promoting activity comprises T/Y-S/N-E-Q-S-C-A. (SEQ ID NO:3).

In some examples, the present invention provides oligopeptides having hair growth promoting activity which comprise at least an amino acid sequence wherein 1 to 3 amino acid residues are substituted in the amino acid sequence for murine pep7 region, Ser-Ile-Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 67), wherein the amino acid residue to be substituted is other than Cys or the amino acid residue to be substituted is other than the third to sixth amino acid residues Glu-Gln-Ser-Cys (SEQ ID NO: 120). In other examples, the present invention provides oligopeptides wherein 0 to 2 amino acid residues are substituted in the amino acid sequence represented by Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 119); wherein the amino acid residue to be substituted is other than Cys or the amino acid residue to be substituted is other than the third to sixth amino acid residues Glu-Gln-Ser-Cys (SEQ ID NO: 120). In some examples, the present invention provides oligopeptides wherein the first Ser is substituted with a hydrophobic amino acid residue or a neutral amino acid residue in the amino acid sequence of Ser-Ile-Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 67).

In other examples, the present invention provides oligopeptides wherein the first Ser is substituted with Ala, Tyr, Thr, Pro, Phe, Val or Gly in the amino acid sequence of Ser-Ile-Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 67). In some examples, the present invention provides oligopeptides wherein the second Ile is substituted with a neutral amino acid residue or a hydrophobic amino acid residue in the amino acid sequence of Ser-Ile-Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 119). In further examples, the present invention provides oligopeptides wherein the second Ile is substituted with Gly, Asn, Thr, Val, Ser, Phe or Leu in the amino acid sequence of Ser-Ile-Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 67). In some examples, the present invention provides oligopeptides wherein the fifth Ser is substituted with a neutral amino acid residue or a hydrophobic amino acid residue in the amino acid sequence of Ser-Ile-Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 67). In other examples, the present invention provides oligopeptides wherein the fifth Ser is substituted with Trp, Phe, Thr, Cys, Tyr, Pro or Ala in the amino acid sequence of Ser-Ile-Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 67). In further examples, the present invention provides oligopeptides wherein the seventh Asp is substituted with a hydrophilic amino acid residue, Gly, Ala or Leu in the amino acid sequence of Ser-Ile-Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 67). The present invention also provides oligopeptides wherein the seventh Asp is substituted with Glu, His, Ser, Ala, Gly, Asn, Tyr or Leu in the amino acid sequence of Ser-Ile-Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 67). In some examples, the present invention provides oligopeptides wherein the third Glu is substituted with Lys, Gly, Gln, Arg, Ala, Val Asp or Trp in the amino acid sequence of Ser-Ile-Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 67). In other examples, the present invention provides oligopeptides wherein the fourth Gln is substituted with Pro, Glu, Thr, Arg, Ser, His or Lys in the amino acid sequence of Ser-Ile-Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 67). In some examples, the present invention provides oligopeptides wherein the first Ser is substituted with Thr or Tyr, the second Ile is substituted with Ser, Asn or Thr, the third Glu is substituted with Ala, Asp or Trp, the fifth Ser is substituted with Cys or Tyr, and/or the seventh Asp is substituted with Gly, Ala or Leu in the amino acid sequence of Ser-Ile-Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 67). In further examples, the present invention provides oligopeptides wherein 1 to 3 amino acid residues other than the third to sixth amino acid residues Glu-Gln-Ser-Cys (SEQ ID NO: 120) are substituted in the amino acid sequence of Ser-Ile-Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 67), the first Ser is substituted with Thr or Tyr, the second Ile is substituted with Ser, Asn or Thr, and/or the seventh Asp is substituted with Gly, Ala or Leu.

In some examples, the present invention provides oligopeptides having hair growth promoting activity which comprise at least an amino acid sequence wherein 1 to 3 amino acid residues are substituted in the amino acid sequence Ser-Ile-Glu-Gln-Cys-Ser-Asp (SEQ ID NO: 67) (a mutation of the murine pep7 region wherein Cys is in the 5th amino acid position of the region) wherein the amino acid residue to be substituted is other than Cys or other than the third to sixth amino acid residues Glu-Gln-Cys-Ser (SEQ ID NO: 120). In further examples, the present invention provides oligopeptides wherein 0 to 2 amino acid residues are substituted in the amino acid sequence represented by Glu-Gln-Cys-Ser-Asp (SEQ ID NO: 112); wherein the amino acid residue to be substituted is other than Cys or other than the third to sixth amino acid residues Glu-Gln-Cys-Ser (SEQ ID NO: 152). In some examples, the present invention provides oligopeptides wherein the first Ser is substituted with a hydrophobic amino acid residue or a neutral amino acid residue in the amino acid sequence of Ser-Ile-Glu-Gln-Cys-Ser-Asp (SEQ ID NO: 4). In other examples, the present invention provides oligopeptides wherein the first Ser is substituted with Ala, Tyr, Thr, Pro, Phe, Val or Gly in the amino acid sequence of Ser-Ile-Glu-Gln-Cys-Ser-Asp (SEQ ID NO: 4). In some examples, the present invention provides oligopeptides wherein the second Ile is substituted with a neutral amino acid residue or a hydrophobic amino acid residue in the amino acid sequence of Ser-Ile-Glu-Gln-Cys-Ser-Asp (SEQ ID NO: 4). In further examples, the present invention provides oligopeptides wherein the second Ile is substituted with Gly, Asn, Thr, Val, Ser, Phe or Leu in the amino acid sequence of Ser-Ile-Glu-Gln-Cys-Ser-Asp (SEQ ID NO: 4). In additional examples, the present invention provides oligopeptides wherein the sixth Ser is substituted with a neutral amino acid residue or a hydrophobic amino acid residue in the amino acid sequence of Ser-Ile-Glu-Gln-Cys-Ser-Asp (SEQ ID NO: 4). In some examples, the present invention provides oligopeptides wherein the sixth Ser is substituted with Trp, Phe, Thr, Cys, Tyr, Pro or Ala in the amino acid sequence of Ser-Ile-Glu-Gln-Cys-Ser-Asp (SEQ ID NO: 4). The present invention also provides oligopeptides wherein the seventh Asp is substituted with a hydrophilic amino acid residue, Gly, Ala or Leu in the amino acid sequence of Ser-Ile-Glu-Gln-Cys-Ser-Asp (SEQ ID NO: 4). In some examples, the present invention provides oligopeptides wherein the seventh Asp is substituted with Glu, His, Ser, Ala, Gly, Asn, Tyr or Leu in the amino acid sequence of Ser-Ile-Glu-Gln-Cys-Ser-Asp (SEQ ID NO: 4). In other examples, the present invention provides oligopeptides wherein the third Glu is substituted with Lys, Gly, Gln, Arg, Ala, Val, Asp or Trp in the amino acid sequence of Ser-Ile-Glu-Gln-Cys-Ser-Asp (SEQ ID NO: 4). In some examples, the present invention provides oligopeptides wherein the fourth Gln is substituted with Pro, Glu, Thr, Arg, Ser, His or Lys in the amino acid sequence of Ser-Ile-Glu-Gln-Cys-Ser-Asp (SEQ ID NO: 4). In other examples, the present invention provides oligopeptides wherein the first Ser is substituted with Thr or Tyr, the second Ile is substituted with Ser, Asn or Thr, the third Glu is substituted with Ala, Asp or Trp, the sixth Ser is substituted with Cys or Tyr, and/or the seventh Asp is substituted with Gly, Ala or Leu in the amino acid sequence of Ser-Ile-Glu-Gln-Cys-Ser-Asp (SEQ ID NO: 4). In some examples, the present invention provides oligopeptides wherein 1 to 3 amino acid residues other than the third to sixth amino acid residues Glu-Gln-Cys-Ser (SEQ ID NO: 152) are substituted in the amino acid sequence of Ser-Ile-Glu-Gln-Cys-Ser-Asp (SEQ ID NO: 4), the first Ser is substituted with Thr or Tyr, the second Ile is substituted with Ser, Asn or Thr, and/or the seventh Asp is substituted with Gly, Ala or Leu.

In other examples, the present invention provides oligopeptides which comprises at least an amino acid sequence wherein 1 to 3 amino acid residues are substituted in the amino acid sequence Ser-Ile-Glu-Cys-Gln-Ser-Asp (SEQ ID NO: 5) (a mutation of the murine pep7 region wherein Cys is in the 4th amino acid position of the region) wherein the amino acid residue to be substituted is other than Cys or wherein the amino acid residue to be substituted is other than the third to sixth amino acid residues Glu-Cys-Gln-Ser (SEQ ID NO: 153). In further examples, the present invention provides an oligopeptide wherein 0 to 2 amino acid residues are substituted in the amino acid sequence represented by Glu-Cys-Gln-Ser-Asp (SEQ ID NO: 113); wherein the amino acid residue to be substituted is other than Cys or wherein the amino acid residue to be substituted is other than the third to sixth amino acid residues Glu-Cys-Gln-Ser (SEQ ID NO: 153). In further examples, the present invention provides oligopeptides wherein the first Ser is substituted with a hydrophobic amino acid residue or a neutral amino acid residue in the amino acid sequence of Ser-Ile-Glu-Cys-Gln-Ser-Asp (SEQ ID NO: 5). In some examples, the present invention provides oligopeptides wherein the first Ser is substituted with Ala, Tyr, Thr, Pro, Phe, Val or Gly in the amino acid sequence of Ser-Ile-Glu-Cys-Gln-Ser-Asp (SEQ ID NO: 5). The present invention further provides oligopeptides wherein the second Ile is substituted with a neutral amino acid residue or a hydrophobic amino acid residue in the amino acid sequence of Ser-Ile-Glu-Cys-Gln-Ser-Asp (SEQ ID NO: 5). In some examples, the present invention provides oligopeptides wherein the second Ile is substituted with Gly, Asn, Thr, Val, Ser, Phe or Leu in the amino acid sequence of Ser-Ile-Glu-Cys-Gln-Ser-Asp (SEQ ID NO: 5). In other examples, the present invention provides oligopeptides wherein the sixth Ser is substituted with a neutral amino acid residue or a hydrophobic amino acid residue in the amino acid sequence of Ser-Ile-Glu-Cys-Gln-Ser-Asp (SEQ ID NO: 5). In some examples, the present invention provides oligopeptides wherein the sixth Ser is substituted with Trp, Phe, Thr, Cys, Tyr, Pro or Ala in the amino acid sequence of Ser-Ile-Glu-Cys-Gln-Ser-Asp (SEQ ID NO: 5). In yet further examples, the present invention provides oligopeptides wherein the seventh Asp is substituted with a hydrophilic amino acid residue, Gly, Ala or Leu in the amino acid sequence of Ser-Ile-Glu-Cys-Gln-Ser-Asp (SEQ ID NO: 5).

In additional examples, the present invention provides oligopeptides wherein the seventh Asp is substituted with Glu, His, Ser, Ala, Gly, Asn, Tyr or Leu in the amino acid sequence of Ser-Ile-Glu-Cys-Gln-Ser-Asp (SEQ ID NO: 5). In some examples, the present invention provides oligopeptides wherein the third Glu is substituted with Lys, Gly, Gln, Arg, Ala, Val, Asp or Trp in the amino acid sequence of Ser-Ile-Glu-Cys-Gln-Ser-Asp (SEQ ID NO: 5). In other examples, the present invention provides oligopeptides wherein the fifth Gln is substituted with Pro, Glu, Thr, Arg, Ser, His or Lys in the amino acid sequence of Ser-Ile-Glu-Cys-Gln-Ser-Asp (SEQ ID NO: 5). In yet other examples, the present invention provides oligopeptides wherein the first Ser is substituted with Thr or Tyr, the second Ile is substituted with Ser, Asn or Thr, the third Glu is substituted with Ala, Asp or Trp, the sixth Ser is substituted with Cys or Tyr, and/or the seventh Asp is substituted with Gly, Ala or Leu in the amino acid sequence of Ser-Ile-Glu-Cys-Gln-Ser-Asp (SEQ ID NO: 5). In other examples, the present invention provides oligopeptides wherein 1 to 3 amino acid residues other than the third to sixth amino acid residues Glu-Cys-Gln-Ser (SEQ ID NO: 153) are substituted in the amino acid sequence of Ser-Ile-Glu-Cys-Gln-Ser-Asp (SEQ ID NO: 5), the first Ser is substituted with Thr or Tyr, the second Ile is substituted with Ser, Asn or Thr, and/or the seventh Asp is substituted with Gly, Ala or Leu.

In some examples, the present invention provides oligopeptides wherein 1 to 3 amino acid residues are substituted in the amino acid sequence represented by Ser-Ile-Cys-Glu-Gln-Ser-Asp (SEQ ID NO: 6) (a mutation of the murine pep7 region wherein Cys is in the 3rd amino acid position of the region) wherein the amino acid residue to be substituted is other than Cys or other than the third to sixth amino acid residues Cys-Glu-Gln-Ser (SEQ ID NO: 154). In further examples, the present invention provides oligopeptides wherein 0 to 2 amino acid residues are substituted in the amino acid sequence represented by Cys-Glu-Gln-Ser-Asp (SEQ ID NO: 114), wherein the amino acid residue to be substituted is other than Cys or other than the third to sixth amino acid residues Cys-Glu-Gln-Ser (SEQ ID NO: 154). In some examples, the present invention provides oligopeptides wherein the first Ser is substituted with a hydrophobic amino acid residue or a neutral amino acid residue in the amino acid sequence of Ser-Ile-Cys-Glu-Gln-Ser-Asp (SEQ ID NO: 6). In some examples, the present invention provides oligopeptides wherein the first Ser is substituted with Ala, Tyr, Thr, Pro, Phe, Val or Gly in the amino acid sequence of Ser-Ile-Cys-Glu-Gln-Ser-Asp (SEQ ID NO: 6). In other examples, the present invention provides oligopeptides wherein the second Ile is substituted with a neutral amino acid residue or a hydrophobic amino acid residue in the amino acid sequence of Ser-Ile-Cys-Glu-Gln-Ser-Asp (SEQ ID NO: 6). In additional examples, the present invention provides oligopeptides wherein the second Ile is substituted with Gly, Asn, Thr, Val, Ser, Phe or Leu in the amino acid sequence of Ser-Ile-Cys-Glu-Gln-Ser-Asp (SEQ ID NO: 6). In other examples, the present invention provides oligopeptides wherein the sixth Ser is substituted with a neutral amino acid residue or a hydrophobic amino acid residue in the amino acid sequence of Ser-Ile-Cys-Glu-Gln-Ser-Asp (SEQ ID NO: 6). In some examples, the present invention provides oligopeptides wherein the sixth Ser is substituted with Trp, Phe, Thr, Cys, Tyr, Pro or Ala in the amino acid sequence of Ser-Ile-Cys-Glu-Gln-Ser-Asp (SEQ ID NO: 6). In further examples, the present invention provides oligopeptides wherein the seventh Asp is substituted with a hydrophilic amino acid residue, Gly, Ala or Leu in the amino acid sequence of Ser-Ile-Cys-Glu-Gln-Ser-Asp (SEQ ID NO: 6). In other examples, the present invention provides oligopeptides wherein the seventh Asp is substituted with Glu, His, Ser, Ala, Gly, Asn, Tyr or Leu in the amino acid sequence of Ser-Ile-Cys-Glu-Gln-Ser-Asp (SEQ ID NO: 6). In additional examples, the present invention provides oligopeptides wherein the fourth Glu is substituted with Lys, Gly, Gln, Arg, Ala, Val, Asp or Trp in the amino acid sequence of Ser-Ile-Cys-Glu-Gln-Ser-Asp (SEQ ID NO: 6). In some examples, the present invention provides oligopeptides wherein the fifth Gln is substituted with Pro, Glu, Thr, Arg, Ser, His or Lys in the amino acid sequence of Ser-Ile-Cys-Glu-Gln-Ser-Asp (SEQ ID NO: 6). In further examples, the present invention provides oligopeptides wherein the first Ser is substituted with Thr or Tyr, the second Ile is substituted with Ser, Asn or Thr, the fourth Glu is substituted with Ala, Asp or Trp, the sixth Ser is substituted with Cys or Tyr, and/or the seventh Asp is substituted with Gly, Ala or Leu in the amino acid sequence of Ser-Ile-Cys-Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 155). The present invention also provides oligopeptides wherein 1 to 3 amino acid residues other than the third to sixth amino acid residues Cys-Glu-Gln-Ser (SEQ ID NO: 154) are substituted in the amino acid sequence of Ser-Ile-Cys-Glu-Gln-Ser-Asp (SEQ ID NO: 6), the first Ser is substituted with Thr or Tyr, the second Ile is substituted with Ser, Asn or Thr, and/or the seventh Asp is substituted with Gly, Ala or Leu.

In some examples, the present invention provides oligopeptides wherein 1 to 3 amino acid residues are substituted in the amino acid sequence represented by Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 111). In other examples, the present invention provides oligopeptides wherein 1 to 3 amino acid residues are substituted in the amino acid sequence represented by Glu-Gln-Cys-Ser-Asp (SEQ ID NO: 112). In some examples, the present invention provides oligopeptides wherein 1 to 3 amino acid residues are substituted in the amino acid sequence represented by Glu-Cys-Gln-Ser-Asp (SEQ ID NO: 113). In other examples, the present invention provides oligopeptides wherein 1 to 3 amino acid residues are substituted in the amino acid sequence represented by Cys-Glu-Gln-Ser-Asp (SEQ ID NO: 114).

In additional examples, the present invention provides oligopeptides comprising an amino acid sequence selected from the group consisting of the following:

Lys-Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu (SEQ ID NO: 83);

Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu (SEQ ID NO: 84);

Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp (SEQ ID NO: 85);

Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln (SEQ ID NO: 86);

Ser-Ile-Glu-Gln-Ser-Cys-Asp (SEQ ID NO: 87);

Ser-Ile-Glu-Gln-Ser-Cys (SEQ ID NO: 88);

Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu (SEQ ID NO: 89);

Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu (SEQ ID NO: 90); and

Gln-Ser-Cys-Asp-Gln-Asp-Glu (SEQ ID NO: 91).

The present invention also provides oligopeptides comprising between about 8 and about 20 amino acid residues and comprising an amino acid sequence selected from the group consisting of:

Ser-Ile-Glu-Gln-Ser-Xaa-Asp-Gln (SEQ ID NO: 115);

Ser-Ile-Glu-Gln-Xaa-Ser-Asp-Gln (SEQ ID NO: 116);

Ser-Ile-Glu-Xaa-Gln-Ser-Asp-Gln (SEQ ID NO: 117); and

Ser-Ile-Xaa-Glu-Gln-Ser-Asp-Gln (SEQ ID NO: 118),

wherein Xaa is Cys, or a reactive substance-bound Cys.

The present invention also provides oligopeptides comprising an amino acid sequence selected from the group consisting of:

Ser-Ile-Glu-Gln-Cys-Ser-Asp-Gln (SEQ ID NO: 92);

Ser-Ile-Glu-Cys-Gln-Ser-Asp-Gln (SEQ ID NO: 93); and

Ser-Ile-Cys-Glu-Gln-Ser-Asp-Gln (SEQ ID NO: 94).

The present invention also provides oligopeptides comprising an amino acid sequence of any one of SEQ ID NO:3 through SEQ ID NO:135. The present invention also provides oligopeptides comprising the following amino acid sequences: Tyr Asn Glu Gln Ser Cys Asp Arg Glu Glu (SEQ ID NO: 17); Thr Ser Asp Gln Cys Cys Asp Pro Asp Lys (SEQ ID NO: 76); Pro Ser Glu Gln Ser Cys Ala Glu Glu Glu (SEQ ID NO: 61); Ser Asn Glu Gln Ser Cys Ala Val Ala Glu (SEQ ID NO: 29); Thr Thr Glu Gln Ser Cys Ala Val Asp Glu (SEQ ID NO: 63); Ser Ile Glu Gin Ser Cys Gly Gln His Glu (SEQ ID NO: 81); Ser Ser Ala Gln Ser Cys Leu Gln Asp Thr (SEQ ID NO: 48); Tyr Ile Glu Gln Tyr Cys Asp Gln Asp Glu (SEQ ID NO: 64); or Thr Ile Trp Gln Ser Cys Asp Gln Glu Glu (SEQ ID NO: 32).

An oligopeptide of the present invention encompasses oligopeptides comprising natural amino acid residues, non-natural amino acid residues, or a mixture of both. An oligopeptide of the present invention encompasses oligopeptides that are modified with a cross-linking agent. The present invention also encompasses oligopeptide polymers comprising at least two oligopeptides of the present invention that are cross-linked, with the proviso that the polymer is not a homopolymer of SEQ ID NO:1 or SEQ ID NO:2. In some examples, the polymer is a dimer, including a homodimer as well as a heterodimer. In other examples, the polymer is a trimer. The present invention encompasses oligopeptide polymers prepared by cross-linking at least two oligopeptides of the present invention, with the proviso that the polymer is not a homopolymer of SEQ ID NO:1 or SEQ ID NO:2.

The present invention encompasses compositions comprising an oligopeptide of the present invention. In some examples, the composition further comprises a pharmaceutically acceptable excipient. In other examples, the composition comprises an agent that enhances transdermal penetration or delivery.

The present invention also provides methods for promoting hair growth in a mammal comprising administering a composition comprising an oligopeptide of the present invention to a mammal in need of hair growth in an amount effective to promote hair growth in said mammal.

The present invention also provides monoclonal antibodies, or fragments thereof, which specifically recognize an antigen of about 220 kDa present in epithelial new follicles. In some examples, the antigen of about 220 kDa present in epithelial new follicles is an antigen which is specifically expressed during the growth period of an imago or the developing period of a fetus. In other examples, the present invention provides a hybridoma deposited with the Patent and Bio-Resource Center of National Institute of Advanced Industrial Science and Technology and having an accession number of FERM P-18578 and a monoclonal antibody made by said hybridoma. In other examples, the present invention provides an antigen, or fragment thereof, recognized by the monoclonal antibody made by the hybridoma deposited with the Patent and Bio-Resource Center of National Institute of Advanced Industrial Science and Technology and having an accession number of FERM P-18578

The present invention also provides a hybridoma produced by the method comprising fusing immunocytes of a mammal immunized with an immunogen containing protein extracted from hair collected from the skin of a mammal in the growth period and/or follicles of whiskers of a mammal in a growth period, and myeloma cells of a mammal. The present invention also provides a process for the production of a monoclonal antibody specific for an antigen of about 220 kDa present in epithelial new follicles, which comprises the steps of incubating the hybridoma deposited with the Patent and Bio-Resource Center of National Institute of Advanced Industrial Science and Technology and having an accession number of FERM P-18578 and collecting the monoclonal antibody produced by said hybridoma.

The present invention also provides methods for the evaluation of hair growth promoting activity comprising the steps of; (1) incubating skin tissue derived from a mammal in the presence of a substance to be tested under suitable conditions and for a time effective to promote hair growth; (2) recovering said skin tissue from step (1); and (3) reacting said skin tissue with the monoclonal antibody made by a hybridoma deposited with the Patent and Bio-Resource Center of National Institute of Advanced Industrial Science and Technology and having an accession number of FERM P-18578 or a fragment thereof; and; (4) detecting said monoclonal antibody or a fragment thereof that reacted with the skin tissue.

The present invention also provides kits comprising a monoclonal antibody made by the hybridoma deposited with the Patent and Bio-Resource Center of National Institute of Advanced Industrial Science and Technology and having an accession number of FERM P-18578

Claim 1 of 42 Claims

1. An isolated oligopeptide of between 6 and 40 amino acid residues in length having hair growth promoting activity, comprising an amino acid sequence selected from the group consisting of the following: TABLE-US-00014 (SEQ ID NO:83, see Original Patent) Lys-Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu; (SEQ ID NO:84) Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu; (SEQ ID NO:85) Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp; (SEQ ID NO:86) Ser-Ile-Glu-Gln-Ser-Cys-Asp-Gln; (SEQ ID NO:87) Ser-Ile-Glu-Gln-Ser-Cys-Asp; (SEQ ID NO:88) Ser-Ile-Glu-Gln-Ser-Cys; (SEQ ID NO:89) Ile-Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu; (SEQ ID NO:90) Glu-Gln-Ser-Cys-Asp-Gln-Asp-Glu; and (SEQ ID NO:91) Gln-Ser-Cys-Asp-Gln-Asp-Glu.

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