High activity growth factor mutants

Abstract

The invention relates to novel biosynthetic growth factor mutants which exhibit improved biological activity.

1 . A method for preventing or treating diseases which are associated with tissue degeneration and/or tissue destruction, in a patient in need thereof, comprising administering to the patient an effective amount of a recombinant protein
 derived from a GDF-5-related protein, or a variant of the GDF-5-related protein, wherein
 a) the amino acid at the position corresponding to methionine 453 (M453) of human wild-type GDF-5 (SEQ ID NO 1) is alanine, valine or isoleucine, and/or 
 b) the amino acid at the position corresponding to methionine 456 (M456) of human wild-type GDF-5 (SEQ ID NO 1) is alanine, valine or isoleucine, 
   wherein the variant is a biologically active fragment, and wherein the GDF-5-related protein is any naturally occurring or artificially created protein which comprises a cysteine-knot domain with an amino acid identity of at least 75% to the 102 amino acid cysteine-knot domain of GDF-5, and further wherein the recombinant protein has an improved biological activity of at least 120% when compared to the mature GDF-5 protein.   
     
     
         2 . A method for the diagnosis, prevention and/or therapy of injuries or diseases associated with damaged bone, cartilage, connective tissue, connective tissue attachment, tendon, ligament, spinal/intervertebral disk, meniscus, dental tissue, dentin, periodontal ligament, blood vessels, skin, hair or neural tissue, in a patient in need thereof, comprising administering to the patient an effective amount of a recombinant protein derived from a GDF-5-related protein, or a variant of the GDF-5-related protein, wherein
 a) the amino acid at the position corresponding to methionine 453 (M453) of human wild-type GDF-5 (SEQ ID NO 1) is alanine, valine or isoleucine, and/or   b) the amino acid at the position corresponding to methionine 456 (M456) of human wild-type GDF-5 (SEQ ID NO 1) is alanine, valine or isoleucine,   
       wherein the variant is a biologically active fragment, and wherein the GDF-5-related protein is any naturally occurring or artificially created protein which comprises a cysteine-knot domain with an amino acid identity of at least 75% to the 102 amino acid cysteine-knot domain of GDF-5, and further wherein the recombinant protein has an improved biological activity of at least 120% when compared to the mature GDF-5 protein. 
     
     
         3 . A method for promoting cartilage and/or bone growth and/or spinal fusion, for the diagnosis, prevention and/or therapy of damaged or diseased tissues selected from the group consisting of tissues of the sensory system, liver, pancreas, cardiac, blood vessel, renal, uterine and thyroid tissue, skin, mucous membranes, endothelium, epithelium; for the induction of nerve growth, tissue regeneration, angiogenesis, wound healing including ulcers, burns, injuries and/or skin grafts; for the induction of proliferation of progenitor cells and/or bone marrow cells; for maintenance of a state of proliferation or differentiation for treatment or preservation of tissue or cells for organ or tissue transplantation; for the treatment of degenerative disorders concerning the joints to skeletal elements and/or for meniscus and/or spinal/intervertebral disk repair; for the manufacture of a therapeutic and/or diagnostic composition for the prevention and/or treatment of neurodegenerative disorders, in a patient in need thereof, comprising administering to the patient an effective amount of a recombinant protein derived from a GDF-5-related protein, or a variant of the GDF-5-related protein, wherein
 a) the amino acid at the position corresponding to methionine 453 (M453) of human wild-type GDF-5 (SEQ ID NO 1) is alanine, valine or isoleucine, and/or   b) the amino acid at the position corresponding to methionine 456 (M456) of human wild-type GDF-5 (SEQ ID NO 1) is alanine, valine or isoleucine,   
       wherein the variant is a biologically active fragment, and wherein the GDF-5-related protein is any naturally occurring or artificially created protein which comprises a cysteine-knot domain with an amino acid identity of at least 75% to the 102 amino acid cysteine-knot domain of GDF-5, and further wherein the recombinant protein has an improved biological activity of at least 120% when compared to the mature GDF-5 protein. 
     
     
         4 . The method according to  claim 3 , wherein said neurodegenerative disorder is selected from the group consisting of Parkinson's disease, Alzheimer's disease, Amyotrophic lateral sclerosis (ALS), Multiple sclerosis and Huntington's disease. 
     
     
         5 . Method according to  claim 4 , wherein said disease associated with bone and cartilage damage is osteoporosis. 
     
     
         6 . The method according to  claim 3 , wherein the damaged or diseased tissue is selected from the group consisting of connective tissue including tendon and/or ligament, periodontal or dental tissue including dental implants, neural tissue including CNS tissue and neuropathological situations, tissue of the sensory system, liver, pancreas, cardiac, blood vessel, renal, uterine and thyroid tissue, skin, mucous membranes, endothelium, epithelium. 
     
     
         7 . The method according to  claim 3 , wherein the tissue is the gastrointestinal lining. 
     
     
         8 . The method according to  claim 3 , wherein the disease is associated with disturbances in fertility, contraception and/or pregnancy. 
     
     
         9 . The method according to  claim 3 , wherein the therapy is associated with promoting hair growth.