Anti-infective therapy

Abstract

DNA isolates coding for human DNase and methods of obtaining such DNA are provided, together with expression systems for recombinant production of human DNase useful in therapeutic or diagnostic compositions.

1 . Human DNase unaccompanied by associated native glycosylation.  
     
     
         2 . A DNA isolate encoding human DNase.  
     
     
         3 . The isolate of  claim 2  wherein the isolate is free of DNase introns.  
     
     
         4 . The isolate of  claim 2  wherein the isolate is free of genomic DNA which encodes another polypeptide from the source of the DNA.  
     
     
         5 . The isolate of  claim 2 , wherein the DNA encodes a polypeptide having the amino sequence shown in FIG. 1.  
     
     
         6 . A recombinant expression vector comprising DNA encoding human DNase.  
     
     
         7 . A composition comprising a cell transformed with the recombinant expression vector of  claim 6 .  
     
     
         8 . The composition of  claim 7  wherein the cell is a mammalian cell.  
     
     
         9 . A process for producing DNase which comprises transforming a host cell with nucleic acid encoding DNase, culturing the transformed cell and recovering DNase from the culture.  
     
     
         10 . The process according to  claim 9  wherein the DNase is recovered from the culture medium of the host cell.  
     
     
         11 . The process according to  claim 10  wherein the host cell is a eukaryotic cell.  
     
     
         12 . The process of  claim 9  wherein the DNase is human DNase.  
     
     
         13 . The process of  claim 12  wherein the eukaryotic cell is a human embryonic kidney cell line.  
     
     
         14 . The process of  claim 12  wherein the nucleic acid encodes a human DNase preprotein.  
     
     
         15 . The process of  claim 14  wherein the preprotein is human preDNase.  
     
     
         16 . The process of  claim 12  wherein the human DNase is secreted into the culture medium.  
     
     
         17 . The process of  claim 12  wherein the human DNase is unglycosylated.  
     
     
         18 . A pharmaceutical preparation useful for enzymatic treatment comprising a therapeutically effective amount of human DNase and a pharmaceutically acceptable carrier.  
     
     
         19 . The preparation of  claim 18  which is sterile.  
     
     
         20 . The preparation of  claim 18  wherein the DNase is entirely free of proteases.  
     
     
         21 . The preparation of  claim 20  in the form of an aerosol.  
     
     
         22 . The preparation of  claim 18  wherein the DNase has the sequence of the mature DNase depicted in FIG. 1.  
     
     
         23 . A polynucleotide probe containing at least about 10 bases which is capable of hybridizing under stringent conditions to the human DNase gene.  
     
     
         24 . A conjugate of human DNase and a nonproteinaceous polymer.  
     
     
         25 . The conjugate of  claim 24  wherein the polymer is surgical tubing selected from the group of catheters and drainage conduits.  
     
     
         26 . The conjugate of  claim 24  which is water soluble.  
     
     
         27 . The conjugate of  claim 24  wherein the polymer is a polyoxyalkylene or polyalkylene glycol.  
     
     
         28 . A method for the treatment of a patient having an accumulation of purulent material comprising administering pure DNase free of proteases to the patient in an amount therapeutically effective to reduce the visco-elasticity of the material.  
     
     
         29 . A method for enhancing the activity of antibiotics comprising administering to a patient a therapeutically effective amount of an antibiotic and a therapeutically effective amount of DNase.  
     
     
         30 . A method for maintaining the flow of fluid in conduits communicating with a patient's body cavity comprising contacting the interior of the conduit with DNase.  
     
     
         31 . The method of  claim 30  wherein the DNase is covalently bound to the conduit.  
     
     
         32 . The method of  claim 30  wherein a solution of the DNase is passed through the conduit into the body cavity.  
     
     
         33 . A method for purifying pharmaceutical preparations so as to be free of contaminant DNA comprising contacting the preparation with DNase under conditions for degrading the contaminant DNA to oligonucleotides and removing the oligonucleotides and DNase from the preparation.  
     
     
         34 . The method of  claim 33  wherein the DNase is immobilized on a water insoluble support.  
     
     
         35 . A method for the treatment of a patient having cystic fibrosis comprising administering to such patient a therapeutically effective dose of pure DNase free of proteases.