Polypeptide

Abstract

The invention relates to a polypeptide, or part thereof, which inhibits the apoptotic activity of the tumour suppressor protein p53 and including screening methods to identify agents which interfere with the activity of said polypeptide.

1 . An isolated nucleic acid molecule which encodes a polypeptide, or sequence variant thereof, wherein said polypeptide is a fragment of the polypeptide sequence represented in SEQ ID NO: 8 or 9, wherein the fragment is: 
 a polypeptide fragment consisting of amino acid residues from about residue 128-224 of the amino acid sequence presented in SEQ ID NO: 8 or 9;    or a polypeptide fragment consisting of amino acid residues from about residue 128-224 of the amino acid sequence presented in SEQ ID NO: 8 or 9 wherein said sequence has been modified by addition, deletion or substitution of at least one amino acid residue, wherein the polypeptide inhibits the apoptoic activity of p53.    
     
     
         2 . The nucleic acid molecule according to  claim 1 , wherein said molecule encodes a fragment consisting of amino acid residues from about residue 128-224 of the sequence represented in SEQ ID NO: 8.  
     
     
         3 . The nucleic acid molecule according to  claim 2 , wherein said molecule is isolated from a human.  
     
     
         4 . The nucleic acid molecule according to  claim 1 , wherein said molecule encodes a fragment consisting of amino acid residues from about residue 128-224 of the sequence represented in SEQ ID NO: 9.  
     
     
         5 . The nucleic acid molecule according to  claim 4 , wherein said molecule is isolated from a nematode.  
     
     
         6 . The nucleic acid molecule according to  claim 5 , wherein said nematode is of the genus  Caenorhabditis  spp.  
     
     
         7 . (canceled)  
     
     
         8 . The nucleic acid molecule according to  claim 1 , wherein said nucleic acid molecule is a cDNA or genomic DNA.  
     
     
         9 . (canceled)  
     
     
         10 . A polypeptide fragment or sequence variant thereof, encoded by the nucleic acid molecule according to  claim 1 .  
     
     
         11 . A vector comprising the nucleic acid according to  claim 1 .  
     
     
         12 . The vector according to  claim 11 , wherein said vector is an expression vector.  
     
     
         13 . A cell tranformed or transfected with the nucleic acid molecule according to  claim 1 .  
     
     
         14 . A pharmaceutical composition comprising the nucleic acid according to  claim 1 .  
     
     
         15 . A pharmaceutical composition comprising the polypeptide according to  claim 10 .  
     
     
         16 . (canceled)  
     
     
         17 . A transgenic non-human animal comprising the nucleic acid molecule according to  claim 1 .  
     
     
         18 . (canceled)  
     
     
         19 . A screening method to identify agents which inhibit the binding of a polypeptide, or fragment thereof, to p53 comprising: 
 forming a preparation comprising 
 the polypeptide of  claim 10;  and  
 a p53 polypeptide, or a fragment thereof consisting of the binding site(s) for the polypeptide of  claim 10;   
   providing at least one agent to be tested; and    determining the activity of the agent with respect to the binding of the polypeptide of  claim 10  to the p53 polypeptide, or a fragment thereof consisting of the binding site(s) for the polypeptide of  claim 10 .    
     
     
         20 . The method according to  claim 19 , wherein said agent is a polypeptide or a peptide.  
     
     
         21 . (canceled)  
     
     
         22 . The method according to  claim 20 , wherein said polypeptide is an antibody or binding part thereof.  
     
     
         23 . The method according to  claim 22 , wherein said antibody is a monoclonal antibody.  
     
     
         24 . The method according to  claim 22 , wherein said fragment is an Fab fragment.  
     
     
         25 . The method according to  claim 24 , wherein said Fab fragment is an F(ab′) 2  fragment, an Fab fragment, an Fv fragment, or CDR3 regions.  
     
     
         26 . The method according to  claim 23 , wherein said antibody is humanised.  
     
     
         27 . The method according to  claim 23 , wherein said antibody is a chimeric antibody.  
     
     
         28 . An isolated nucleic acid molecule, wherein said molecule is isolated from a nematode worm and hybridises to the nucleic acid sequence shown in SEQ ID NO: 9, wherein said nucleic acid molecule encodes an inhibitor of p53 and inhibits the apoptotic activity of p53.  
     
     
         29 . The nucleic acid molecule according to  claim 28 , wherein said molecule hybridises under stringent hybridisation conditions.  
     
     
         30 . (canceled)  
     
     
         31 . An isolated polypeptide comprising the amino acid sequence as represented in SEQ ID NO: 9 or a variant polypeptide which polypeptide is modified by addition, deletion or substitution of at least one amino acid residue and is an inhibitor of p53.  
     
     
         32 . A method of treatment of an animal, comprising administering an effective amount of the polypeptide according to  claim 10 , wherein said effective amount induces the apoptotic activity of p53.  
     
     
         33 . A method of treatment of an animal comprising administering an effective amount of a nucleic acid molecule according to  claim 1 , wherein said effective amount induces the apoptotic activity of p53.  
     
     
         34 . The method according to  claim 32 , wherein said treatment is of cancer.  
     
     
         35 . The polypeptide of  claim 10 , wherein the polypeptide is a peptide comprising the amino acid sequence DGPEETD (SEQ ID NO: 2); GPEETD (SEQ ID NO: 2); TTLSDG (SEQ ID NO: 3); AEFGDE (amino acids 294-299 of SEQ ID NO: 8); or PRNYFG (SEQ ID NO: 4).  
     
     
         36 . The peptide according to  claim 35 , wherein said peptide is at least 6 amino acid residues.  
     
     
         37 . The peptide according to  claim 35 , wherein said peptide is at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acid residues.  
     
     
         38 . The peptide according to  claim 35 , wherein said peptide is at least 20; 30; 40; 50; 60; 70; 80; 90; or 100 amino acid residues.  
     
     
         39 . The peptide according to  claim 35 , wherein the peptide consists of the amino acid sequence DGPEETD (SEQ ID NO: 2); GPEETD (SEQ ID NO: 1); TTLSDG (SEQ ID NO: 3); AEFGDE (amino acids 294-299 of SEQ ID NO: 8); or PRNYFG (SEQ ID NO: 4).  
     
     
         40 . (canceled)  
     
     
         41 . The peptide according to  claim 35 , wherein the peptide further comprises at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 arginine residues.  
     
     
         42 . (canceled)  
     
     
         43 . A pharmaceutical composition comprising the peptide of  claim 35 .  
     
     
         44 . The pharmaceutical composition according to  claim 43 , wherein said composition further includes a carrier, diluent or excipient.  
     
     
         45 . The pharmaceutical composition of  claim 35  further comprising at least one anti-cancer agent.  
     
     
         46 . The pharmaceutical composition according to  claim 45  wherein said anticancer agent is cisplatin; carboplatin; cyclosphosphamide; melphalan; carmusline; methotrexate; 5-fluorouracil; cytarabine; mercaptopurine; daunorubicin; doxorubicin; epirubicin; vinblastine; vincristine; dactinomycin; mitomycin C; taxol; L-asparaginase; G-CSF; etoposide; colchicine; derferoxamine mesylate; or camptothecin.  
     
     
         47 . (canceled)  
     
     
         48 . (canceled)  
     
     
         49 . A complex comprising the peptide according to  claim 35  and an antibody, or binding part thereof.  
     
     
         50 . The complex according to  claim 49 , wherein said antibody or binding part is a cell specific antibody.  
     
     
         51 . The complex according to  claim 49  wherein said antibody is a cancer cell specific antibody.  
     
     
         52 . A method of treatment of an animal that would benefit from the induction of apoptosis, comprising administering an effective amount of the peptide of  claim 35  to the animal.  
     
     
         53 . (canceled)  
     
     
         54 . The method according to  claim 52 , wherein said treatment is cancer treatment.