Human DNase II

Abstract

This invention relates to a novel human deoxyribonuclease, referred to as human DNase II. The invention provides nucleic acid sequences encoding human DNase II, thereby enabling the production of human DNase II by recombinant DNA methods in quantities sufficient for clinical use. The invention also relates to pharmaceutical compositions and diagnostic and therapeutic uses of human DNase II.

1 . An isolated nucleic acid molecule comprising a nucleotide sequence encoding the amino acid sequence shown in  FIG. 1  for mature human DNase II.  
     
     
         2 . An expression vector comprising a nucleotide sequence of  claim 1  encoding the amino acid sequence shown in  FIG. 1  for mature human DNase II operably linked to a promoter recognized by a host cell transformed with the vector.  
     
     
         3 . The isolated nucleic acid molecule of  claim 1  comprising a nucleotide sequence that encodes an amino acid sequence having at least 95% identity with the amino acid sequence shown in  FIG. 1  for mature human DNase II.  
     
     
         4 . The isolated nucleic acid molecule of  claim 3  comprising a nucleotide sequence that encodes an amino acid sequence that differs from the amino acid sequence shown in  FIG. 1  for mature human DNase II by the substitution of one amino acid for another at only a single position within the  FIG. 1  sequence.  
     
     
         5 . The isolated nucleic acid molecule of  claim 3  comprising a nucleotide sequence that encodes an amino acid sequence that differs from the amino acid sequence shown in  FIG. 1  for mature human DNase II by the substitution of one amino acid for another at only two positions within the  FIG. 1  sequence.  
     
     
         6 . A host cell transformed with an expression vector of  claim 2  comprising a nucleotide sequence encoding the amino acid sequence shown in  FIG. 1  for mature human DNase II.  
     
     
         7 . A method for producing human DNase II comprising: 
 (a) transforming a cell containing an endogenous human DNase II gene with a homologous DNA comprising an amplifiable gene and a flanking sequence of at least about 150 base pairs that is homologous with a DNA sequence within or in proximity to the DNase II gene, whereby the homologous DNA integrates into the cell genome by recombination;    (b) culturing the cells under conditions that select for amplification of the amplifiable gene, whereby the human DNase II gene is also amplified; and thereafter    (c) recovering human DNase II from the cells.    
     
     
         8 . An isolated polypeptide comprising the amino acid sequence shown in  FIG. 1  for mature human DNase II.  
     
     
         9 . An isolated polypeptide comprising an amino acid sequence having at least 95% identity with the amino acid sequence shown in  FIG. 1  for mature human DNase II, which polypeptide has DNA-hydrolytic activity.  
     
     
         10 . The isolated polypeptide of  claim 9  comprising an amino acid sequence that differs from the amino acid sequence shown in  FIG. 1  for mature human DNase II by the substitution of one amino acid for another at only a single position within the  FIG. 1  sequence.  
     
     
         11 . A polypeptide of  claim 12  wherein the amino acid substitution creates a glycosylation site within the polypeptide that is not present in naturally-occurring human DNase II.  
     
     
         12 . An antibody that is capable of binding to the amino acid sequence shown in  FIG. 1  for mature human DNase II.  
     
     
         13 . The antibody of  claim 16  that is a monoclonal antibody.  
     
     
         14 . A method for the treatment of a patient having a pulmonary disease or disorder comprising administering to the patient a therapeutically effective amount of human DNase II according to  claim 9 .  
     
     
         15 . The method of claim  18  wherein the disease or disorder is cystic fibrosis.  
     
     
         16 . A method for the treatment of a patient having systemic lupus erythematosus comprising administering to the patient a therapeutically effective amount of human DNase II according to  claim 9.