Tgf-beta3 mutants

Abstract

The invention provides TGF-β3s, or fragments or derivatives thereof, wherein the alpha-helix-forming domain between amino acid residues (58) and (67) of full-length wild type TGF-β3 comprises at least one alpha-helix-stabilising substitution. The invention also provides TGF-β3s, or fragments or derivatives thereof, wherein the Glycine residue at position (63) of full-length wild type TGF-β3 is replaced with Proline. Further still, the invention provides TGF-β3s, or fragments or derivatives thereof, comprising a substitution of the Glutamic acid residue at position (12) of full-length wild type TGF-β3 and/or the Arginine residue at position (52) of full-length wild type TGF-β3. The invention also provides medicaments and methods of treatment using such TGF-β3s.

1 . An isolated TGF-β3 protein, or a fragment or derivative thereof, wherein an alpha-helix-forming domain between amino acid residues 58 and 67 of full-length wild type TGF-β3 comprises at least one alpha-helix-stabilising substitution. 
     
     
         2 . An isolated TGF-β3 protein, or fragment or derivative thereof, according to  claim 1 , wherein a glycine residue at position 63 of full-length wild type TGF-β3 is replaced with an alpha-helix-stabilising amino acid residue. 
     
     
         3 . An isolated TGF-β3 protein, or fragment or derivative thereof, according to  claim 1 , wherein the alpha-helix-stabilising substitution comprises substitution of a residue selected from the group consisting of an Alanine, a Serine, a Threonine, a Valine, a Leucine, an Isoleucine, a Methionine and a Phenylalanine residue at a position corresponding to the position of full-length wild type TGF-β3 as set forth in SEQ ID No: 1. 
     
     
         4 . An isolated TGF-β3 protein, or a fragment or derivative thereof, according to  claim 2 , wherein the Glycine residue at position 63 of full-length wild type TGF-β3 is substituted with an alanine residue. 
     
     
         5 . An isolated TGF-β3 protein, according to  claim 4  comprising Sequence ID No. 3, or a fragment or derivative thereof. 
     
     
         6 . An isolated TGF-β3 protein, or a fragment or derivative thereof, according to  claim 1 , which does not comprise a substitution of a Valine residue at position 61 of full-length wild type TGF-β3. 
     
     
         7 . An isolated TGF-β3 protein, or a fragment or derivative thereof, wherein a Glycine residue at position 63 of full-length wild type TGF-β3 is substituted with a Proline residue. 
     
     
         8 . An isolated TGF-β3 protein according to  claim 7  comprising Sequence ID No. 5, or a fragment or derivative thereof. 
     
     
         9 . An isolated nucleic acid encoding a TGF-β3, or fragment or derivative thereof, according to  claim 1 . 
     
     
         10 . A method of accelerating wound healing, or preventing reducing or inhibiting scarring, the method comprising providing administering a therapeutically effective amount of a TGF-β3, or a fragment or derivative thereof, according to  claim 1 , to a patient in need thereof. 
     
     
         11 . A method of preventing or treating a fibrotic disorder, the method comprising providing administering a therapeutically effective amount of a TGF-β3, or a fragment or derivative thereof, according to  claim 1 , to a patient in need. 
     
     
         12 . A method according to  claim 10 , wherein the TGF-β3, or fragment or derivative thereof, is administered to the skin of the patient in need thereof. 
     
     
         13 . A method according to  claim 10 , wherein the TGF-β3, or fragment or derivative thereof, is administered to the eye of the patient in need thereof. 
     
     
         14 . A method according to  claim 11 , wherein the fibrotic disorder is selected from the group consisting of lung fibrosis, liver fibrosis, scleroderma, skin fibrosis, muscle fibrosis, radiation fibrosis, kidney fibrosis, proliferative vitreoretinopathy, and uterine fibrosis. 
     
     
         15 . A method of promoting epithelial regeneration, the method comprising administering a therapeutically effective amount of a TGF-β3, or a fragment or derivative thereof, according to  claim 1 , to a patient in need thereof. 
     
     
         16 . A method of treating a disorder or condition, the method comprising:
 administering a therapeutically effective amount of a TGF-β3, or a fragment or derivative thereof, according to  claim 1 , to a patient in need thereof, and   wherein the disorder or condition is selected from the group consisting of angiogenic disorders, restenosis, adhesions, endometriosis, ischemic disease, oral mucositis, renal disease, bone and cartilage induction, and in vitro fertilisation.   
     
     
         17 . A composition comprising a TGF-β3, or a fragment or derivative thereof, according to  claim 1 , and a pharmaceutically acceptable vehicle. 
     
     
         18 . A composition comprising a TGF-β3, or a fragment or derivative thereof, according to  claim 4 , and a pharmaceutically acceptable vehicle. 
     
     
         19 . A composition comprising a TGF-β3, or a fragment or derivative thereof, according to  claim 7 , and a pharmaceutically acceptable vehicle. 
     
     
         20 . The isolated TGF-β3 protein of  claim 4 , wherein the substituted position 63 corresponds to the position of full-length wild type TGF-β3 as set forth in SEQ ID No: 1. 
     
     
         21 . The isolated TGF-β3 protein of  claim 7 , wherein the substituted position 63 corresponds to the position of full-length wild type TGF-β3 as set forth in SEQ ID No: 1. 
     
     
         22 . An isolated nucleic acid encoding a TGF-β3, or fragment or derivative thereof, wherein the TGF-β3 protein comprises a substitution or substitutions at the amino acid residues selected from the group consisting of (a) the glutamic acid residue at position 12, (b) the arginine residue at position 52, and (c) both the glutamic acid residue at position 12 and the arginine residue at position 52, wherein each of the substituted positions corresponds to the position in full-length wild type TGF-β3 as set forth in SEQ ID NO: 1. 
     
     
         23 . A method of accelerating wound healing, or preventing reducing or inhibiting scarring, the method comprising providing a therapeutically effective amount of a TGF-β3, or a fragment or derivative thereof, to a patient in need thereof, wherein the TGF-β3 protein comprises a substitution or substitutions at the amino acid residues selected from the group consisting of (a) the glutamic acid residue at position 12, (b) the arginine residue at position 52, and (c) both the glutamic acid residue at position 12 and the arginine residue at position 52, wherein each of the substituted positions corresponds to the position in full-length wild type TGF-β3 as set forth in SEQ ID NO: 1. 
     
     
         24 . A method of preventing or treating a fibrotic disorder, the method comprising providing a therapeutically effective amount of a TGF-β3, or a fragment or derivative thereof, to a patient in need thereof, wherein the TGF-β3 protein comprises a substitution or substitutions at the amino acid residues selected from the group consisting of (a) the glutamic acid residue at position 12, (b) the arginine residue at position 52, and (c) both the glutamic acid residue at position 12 and the arginine residue at position 52, wherein each of the substituted positions corresponds to the position in full-length wild type TGF-β3 as set forth in SEQ ID NO: 1. 
     
     
         25 . A method of promoting epithelial regeneration, the method comprising administering a therapeutically effective amount of a TGF-β3, or a fragment or derivative thereof, to a patient in need thereof, wherein the TGF-β3 protein comprises a substitution or substitutions at the amino acid residues selected from the group consisting of (a) the glutamic acid residue at position 12, (b) the arginine residue at position 52, and (c) both the glutamic acid residue at position 12 and the arginine residue at position 52, wherein each of the substituted positions corresponds to the position in full-length wild type TGF-β3 as set forth in SEQ ID NO: 1. 
     
     
         26 . A method of treating a disorder or condition, the method comprising:
 administering a therapeutically effective amount of a TGF-β3, or a fragment or derivative thereof, to a patient in need thereof, wherein the TGF-β3 protein comprises a substitution or substitutions at the amino acid residues selected from the group consisting of (a) the glutamic acid residue at position 12, (b) the arginine residue at position 52, and (c) both the glutamic acid residue at position 12 and the arginine residue at position 52, wherein each of the substituted positions corresponds to the position in full-length wild type TGF-β3 as set forth in SEQ ID NO: 1, and   wherein the disorder or condition is selected from the group consisting of angiogenic disorders, restenosis, adhesions, endometriosis, ischemic disease, oral mucositis, renal disease, bone and cartilage induction, and in vitro fertilisation.