Human DNase

Abstract

This invention relates to a novel human deoxyribonuclease, referred to as LS-DNase, that is relatively resistant to inhibition by actin, as compared to human DNase I. The invention provides nucleic acid sequences encoding LS-DNase, thereby enabling the production of LS-DNase by recombinant DNA methods in quantities sufficient for clinical use. The invention also relates to pharmaceutical compositions and therapeutic uses of LS-DNase.

1 . An isolated nucleic acid molecule comprising a nucleotide sequence encoding the amino acid sequence shown in  FIG. 1  for mature LS-DNase.  
     
     
         2 . An expression vector comprising a nucleotide sequence encoding the amino acid sequence shown in  FIG. 1  for mature LS-DNase operably linked to a promoter recognized by a host cell transformed with the vector.  
     
     
         3 . An isolated nucleic acid molecule 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 LS-DNase.  
     
     
         4 . An isolated nucleic acid molecule comprising a nucleotide sequence that encodes an amino acid sequence that differs from the amino acid sequence shown in  FIG. 1  for mature LS-DNase by the substitution of one amino acid for another at only a single position within the  FIG. 1  sequence.  
     
     
         5 . An isolated nucleic acid molecule comprising a nucleotide sequence that encodes an amino acid sequence that differs from the amino acid sequence shown in  FIG. 1  for mature LS-DNase 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 comprising a nucleotide sequence encoding the amino acid sequence shown in  FIG. 1  for mature LS-DNase.  
     
     
         7 . A method of using a host cell transformed with an expression vector comprising a nucleotide sequence encoding the amino acid sequence shown in  FIG. 1  for mature LS-DNase, which comprises culturing the host cell under conditions such that expression vector is replicated.  
     
     
         8 . A process which comprises transforming a host cell with a nucleic acid molecule that encodes a polypeptide comprising the amino acid sequence shown in  FIG. 1  for mature LS-DNase and culturing the host cell under conditions such that the polypeptide is produced in the host cell.  
     
     
         9 . A method for producing LS-DNase comprising: 
 (a) transforming a cell containing an endogenous LS-DNase 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 endogenous AL-1 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 LS-DNase gene is also amplified; and thereafter    (c) recovering LS-DNase from the cells.    
     
     
         10 . An isolated polypeptide comprising the amino acid sequence shown in  FIG. 1  for mature LS-DNase.  
     
     
         11 . An isolated polypeptide comprising an amino acid sequence having at least 95% identity with the amino acid sequence shown in  FIG. 1  for mature LS-DNase, which polypeptide has DNA-hydrolytic activity.  
     
     
         12 . An isolated polypeptide comprising an amino acid sequence that differs from the amino acid sequence shown in  FIG. 1  for mature LS-DNase by the substitution of one amino acid for another at only a single position within the  FIG. 1  sequence.  
     
     
         13 . A polypeptide of  claim 12  wherein the amino acid substitution creates a glycosylation site within the polypeptide that is not present in human LS-DNase.  
     
     
         14 . A pharmaceutical composition comprising a polypeptide comprising the amino acid sequence shown in  FIG. 1  for mature LS-DNase and a physiologically acceptable excipient.  
     
     
         15 . A composition of  claim 14  that is sterile.  
     
     
         16 . An antibody that is capable of binding to the amino acid sequence shown in  FIG. 1  for mature LS-DNase.  
     
     
         17 . An antibody of  claim 16  that is a monoclonal antibody.  
     
     
         18 . A method for the treatment of a patient having a pulmonary disease or disorder comprising administering to the patient a therapeutically effective amount of LS-DNase.  
     
     
         19 . The method of  claim 18  wherein the disease or disorder is cystic fibrosis.  
     
     
         20 . A method for the treatment of a patient having systemic lupus erythematosus comprising administering to the patient a therapeutically effective amount of LS-DNase.