Aggrecanase molecules

Abstract

Novel aggrecanase proteins and the nucleotide sequences encoding them as well as processes for producing them are disclosed. Methods of identifying and developing inhibitors of the aggrecanase enzymes and antibodies to the enzymes for treatment of conditions characterized by the degradation of aggrecan are also disclosed.

1 . An isolated DNA molecule comprising a DNA sequence chosen from: 
 a) the sequence of SEQ ID NO: 1 from nucleotide #1-#3663;    b) fragments of SEQ ID NO: 1;    c) variants of SEQ ID NO: 1;    d) sequences which hybridize under stringent conditions with SEQ ID NO: 1; and    e) naturally occurring human allelic sequences and equivalent degenerative codon sequences of (a) to (d).    
     
     
         2 . A vector comprising a DNA molecule of  claim 1  in operative association with an expression control sequence therefor.  
     
     
         3 . A host cell transformed with the DNA sequence of  claim 1 .  
     
     
         4 . A host cell transformed with a DNA sequence of  claim 2 .  
     
     
         5 . A method for producing an isolated human aggrecanase protein, said method comprising: 
 a) culturing a host cell transformed with a DNA molecule according to  claim 1;  and    b) recovering and purifying said aggrecanase protein encoded by the DNA molecule from the culture medium.    
     
     
         6 . The method of  claim 5 , wherein said host cell is an insect cell.  
     
     
         7 - 8 . (canceled)  
     
     
         9 . An antibody that binds to an isolated aggrecanase protein comprising an amino acid sequence chosen from: 
 a) the amino acid sequence of SEQ ID NO: 2 from amino acid #1-#1221;    b) fragments of SEQ ID NO: 2; and    c) variants of aggrecanase proteins consisting of addition, substitution, and deletion mutants of the sequences of (a) to (b).    
     
     
         10 . The antibody of  claim 9 , wherein the antibody inhibits aggrecanase activity.  
     
     
         11 . A method for identifying inhibitors of aggrecanase comprising 
 a) providing an aggrecanase protein chosen from SEQ ID NO: 2 or a fragment thereof;    b) combining the aggrecanase protein with a potential inhibitor; and    c) evaluating whether the potential inhibitor inhibits aggrecanase activity.    
     
     
         12 . The method of  claim 11  wherein the method further comprises evaluating the aggrecanase protein in a three dimensional structural analysis prior to combining with the potential inhibitor.  
     
     
         13 . The method of  claim 11  wherein the method further comprises evaluating the aggrecanase protein in a computer aided drug design program prior to combining with the potential inhibitor.  
     
     
         14 . A pharmaceutical composition for inhibiting the proteolytic activity of aggrecanase, wherein the composition comprises an antibody according to  claim 9  and a pharmaceutical carrier.  
     
     
         15 . A method for inhibiting aggrecanase in a mammal comprising administering to said mammal an amount of the composition of  claim 14  effective to inhibit aggrecanase activity.  
     
     
         16 . The method of  claim 15 , wherein the composition is administered intravenously, subcutaneously, or intramuscularly.  
     
     
         17 . The method of  claim 15 , wherein the composition is administered at a dosage of about 500 μg/kg to about 1 mg/kg.  
     
     
         18 . The DNA molecule of  claim 1 , wherein the DNA molecule comprises nucleotides 1-3663 of SEQ ID NO:1.