Peptides That Stimulate Cell Survival and Axon Regeneration

Abstract

Peptides which consist of or comprise the tetrameric peptide structural unit: Xaa-Xaa-Xaa-Xaa (SEQ.ID.NO.: 1) in which Xaa at position 1 represents Glu or Asp, Xaa at position 2 represents any amino acid, Xaa at position 3 represents any amino acid and Xaa at position 4 represents Glu or Asp, each of the meanings of Xaa being independent, and peptides which consist of or comprise the sequence PYSSTA, particularly when in multimeric form, mimic the beneficial trophic and neuritogenic effects of FGF but lack the undesirable mitogenic effects. They are useful for the treatment of conditions for which FGF has been proposed, including treatment of neurodegenerative diseases, ischaemia, wound healing and stimulation of angiogenesis in cardiac muscle.

1 . A purified peptide of from 4 to 30 amino acids, that comprises or consists of the sequence of DRVE, EGME, EMGE, DRSE, DAVE, EVRD, EGGE, DOVE, DXVE, DRXE, DXVD, DRXD, EXVE, ERXE, EXVD, or ERXD, wherein X is any amino acid.  
     
     
         2 .- 65 . (canceled)  
     
     
         66 . The peptide of  claim 1 , further comprising the sequence PYSSTA or four or five contiguous amino acids of the sequence PYSSTA.  
     
     
         67 . The peptide of  claim 1 , comprising or consisting of the sequence of DXXEPYSSTA, DXXDPYSSTA, EXXDPYSSTA, or EXXEPYSSTA, wherein X is any amino acid.  
     
     
         68 . The peptide of  claim 1 , comprising or consisting of the sequence DRVEPYSSTA, DRSEPYSSTA, DAVEPYSSTA, or the corresponding sequences having a cysteine residue at each terminus.  
     
     
         69 . The peptide of  claim 1 , wherein said peptide comprises one or more modifications selected from the group consisting of an acyl group at the N-terminus, an amide group at the carboxy terminus, and side chain modifications.  
     
     
         70 . The peptide of  claim 69 , wherein said peptide has the structure acetyl-DRVEPYSSTA-amide (Peptide A), acetyl-DRVE-amide (Peptide B), acetyl-EGME-amide (Peptide C), acetyl-EGMEGM-amide (Peptide D), acetyl-DRSEPYSSTA-amide (Peptide E) or acetyl-DAVEPYSSTA-amide (Peptide F).  
     
     
         71 . The peptide of  claim 69 , wherein said peptide is a modified peptide attached to a linker group.  
     
     
         72 . The peptide of  claim 71 , wherein said peptide has the structure acetyl-DRVEPYSSTA-X, acetyl-DRVE-X, acetyl-EGME-X, acetyl-EMGE-X, acetyl-DRSE-X, acetyl-DAVE-X, acetyl-EVRD-X, acetyl-EGGE-X, acetyl-EGMEGM-X, acetyl-DRSEPYSSTA-X or acetyl-DAVEPYSSTA-X, wherein X is a linker group.  
     
     
         73 . The peptide of  claim 1 , wherein said peptide is in multimeric form, linked to one or more peptides in accordance with  claim 1  via a backbone structure.  
     
     
         74 . The peptide of  claim 73 , wherein said peptide in multimeric form has the structure [{Ac-DRVEPYSSTA} 2 -K] 2 -K—OH (Peptide A(d)), wherein Ac is an acyl group.  
     
     
         75 . The peptide of  claim 74 , wherein said acyl group is a lower acyl group of from 1 to 4 carbon atoms.  
     
     
         76 . The peptide of claim  730 , wherein said peptide in multimeric form comprises the structure acetyl-DRVEPYSSTA-(X) n , acetyl-DRVE-(X) n , acetyl-EGME-(X) n , acetyl-EMGE-(X) n , acetyl-DRSE-(X) n , acetyl-DAVE-(X) n , acetyl-EVRD-(X) n , acetyl-EGGE-(X) n , acetyl-EGMEGM-(X) n , acetyl-DRSEPYSSTA-(X) n  or acetyl-DAVEPYSSTA-(X) n , linked to a backbone structure, wherein X is a linker group and n is an integer of two or more.  
     
     
         77 . The peptide of  claim 1 , comprising two cysteine residues, wherein said peptide may be cyclized by formation of a disulfide bond between said two cysteine residues.  
     
     
         78 . The peptide of  claim 1 , comprised in a pharmaceutical composition in admixture with a pharmaceutically-suitable carrier.  
     
     
         79 . The peptide of  claim 78 , comprised in a pharmaceutical composition that is formulated to cross the blood-brain barrier.  
     
     
         80 . The peptide of  claim 78 , comprised in a pharmaceutical composition that is formulated for oral, intravenous or topical administration or for application to a wound.  
     
     
         81 . A purified peptide that consists essentially of the first fibronectin type III repeat of NCAM.  
     
     
         82 . An isolated nucleic acid that encodes the peptide of  claim 1 , or that encodes a peptide having an amino acid sequence that is the reverse of the amino acid sequence defined in  claim 1 .  
     
     
         83 . A method for producing a purified peptide, comprising: preparing a peptide in accordance with  claim 1 , by culturing a host cell that comprises an isolated nucleic acid that encodes the peptide of  claim 1  under conditions effective to express said peptide, or by chemically synthesizing a peptide in accordance with  claim 1;  purifying said peptide; and optionally modifying said peptide with one or more modifications selected from the group consisting of an acyl group at the N-terminus, an amide group at the carboxy terminus, and side chain modifications, or linking two or more of said peptides to a linker group or a backbone structure to form a peptide multimer.  
     
     
         84 . A method for stimulating cell survival in a mammal, comprising administering to a mammal in need thereof a biologically-effective amount of the purified peptide of  claim 1 .  
     
     
         85 . The method of  claim 84 , wherein said peptide stimulates survival of neurons, oligodendrocytes or fibroblasts in said mammal.  
     
     
         86 . The method of  claim 84 , wherein said mammal has a neurodegenerative disease, multiple sclerosis, a peripheral neuropathy or is in need of stimulating or restoring nerve function after trauma or surgery.  
     
     
         87 . The method of  claim 86 , wherein said mammal has motor neuron disease, multiple sclerosis, Alzheimer's disease, Parkinson's disease, progressive supranuclear palsy (PSP), a prion disease, diabetic neuropathy, chemotherapy-induced neuropathy, nerve injury, paralysis or spinal cord injury.  
     
     
         88 . The method of  claim 84 , wherein said peptide promotes wound healing in said mammal.  
     
     
         89 . The method of  claim 87 , wherein said mammal is in need of healing wounds caused by trauma, surgery, thermal burns, chemical burns, radiation burns, radiation damage or radiotherapy.  
     
     
         90 . The method of  claim 84 , wherein said peptide stimulates angiogenesis in cardiac muscle in said mammalian.  
     
     
         91 . A method for treating a pathological condition for which fibroblast growth factor (FGF) has been used or proposed for use, comprising administering to a mammal in need thereof a biologically effective amount of the purified peptide of  claim 1 .  
     
     
         92 . A method for inhibiting an undesirable effect of FGF in a mammal, comprising administering to a mammal in need thereof an amount of the purified peptide of  claim 1  effective to inhibit said undesirable effect.  
     
     
         93 . A method for stimulating or restoring nerve function after trauma or surgery in a mammalian subject, comprising administering to a subject in need of such treatment an amount of the purified peptide of  claim 1  effective to stimulate or restore said nerve function.  
     
     
         94 . A method of stimulating angiogenesis in cardiac muscle in a mammalian subject, comprising administering to a subject in need of such treatment an amount of the peptide of  claim 1  effective to stimulate said angiogenesis.  
     
     
         95 . A method of treatment of ischemia in a mammalian subject, comprising administering to a subject in need of such treatment an amount of the peptide of  claim 1  effective for said treatment.  
     
     
         96 . A method of inhibiting or reducing angiogenesis in a tumor in a mammalian subject, comprising administering to a subject in need of such treatment an amount of the peptide of  claim 1  effective for inhibiting or reducing said angiogenesis.