Methods for sterilizing preparations of digestive enzymes

Abstract

Methods are disclosed for sterilizing preparations of digestive enzymes to reduce the level of one or more active biological contaminants or pathogens therein, such as viruses, bacteria (including inter- and intracellular bacteria, such as mycoplasmas, ureaplasmas, nanobacteria, chlamydia, rickettsias), yeasts, molds, fungi, prions or similar agents responsible, alone or in combination, for TSEs and/or single or multicellular parasites. These methods involve sterilizing preparations of digestive enzymes, such as trypsin, α-galactosidase and iduronate-2-sulfatase, with irradiation.

1 . (canceled)  
     
     
         2 . A method for sterilizing a preparation of one or more digestive enzymes that is sensitive to radiation, said method comprising: 
 (i) adding to said preparation at least one stabilizer in an amount effective to protect said preparation of one or more digestive enzymes from said radiation; and    (ii) irradiating said preparation of one or more digestive enzymes with gamma radiation at an effective rate for a time effective to sterilize said preparation of one or more digestive enzymes.    
     
     
         3 - 85 . (canceled)  
     
     
         86 . The method according to  claim 2 , wherein the at least one stabilizer is selected from the group consisting of ascorbic acid or a salt or ester thereof, uric acid or a salt or ester thereof, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid, glutathione, methionine, histidine, N-acetyl cysteine, mannitol, trehalose and dimethyl sulfoxide (DMSO).  
     
     
         87 . The method according to  claim 86 , wherein the at least one stabilizer is at least two stabilizers selected from the group consisting of mannitol, trehalose and DMSO.  
     
     
         88 . The method according to  claim 87 , wherein the at least two stabilizers are mannitol and trehalose.  
     
     
         89 . The method according to  claim 2 , wherein the effective rate is at least 3.0 kGy/hour.  
     
     
         90 . The method according to  claim 2 , wherein the total dose of radiation is at least about  25  kGy.  
     
     
         91 . The method according to  claim 2 , wherein the total dose of radiation is at least about 45 kGy.  
     
     
         92 . The method according to  claim 2 , wherein the one or more digestive enzymes is selected from the group consisting of pancreatic enzymes, salivary enzymes, intestinal enzymes, glycosidases and sulfatases.  
     
     
         93 . The method according to  claim 92 , whrerein the pancreatic enzymes are selected from the group consisting of trypsin, chymotrypsin, pancreatic lipase and pancreatic amylase.  
     
     
         94 . The method according to  claim 92 , wherein the salivary enyme is ptyalin.  
     
     
         95 . The method according to  claim 92 , wherein the intestinal enzymes are selected from the group consisting of intestinal polypeptidases, intestinal amylases and intestinal lipases.  
     
     
         96 . The method according to  claim 92 , wherein the glycosidase is α-galactosidase.  
     
     
         97 . The method according to  claim 92 , wherein the sulfatase is iduronodate-2-sulfatase.  
     
     
         98 . The method according to  claim 2 , wherein the preparation of one or more digestive enzymes is at a temperature below its freezing point during irradiation.  
     
     
         99 . The method according to  claim 2 , wherein the preparation of one or more digestive enzymes is at a temperature below about −20° C.  
     
     
         100 . The method according to  claim 2 , wherein the preparation of one or more digestive enzymes is at a temperature below about −60° C.  
     
     
         101 . The method according to  claim 2 , wherein the preparation of one or more digestive enzymes is at a temperature below about −78° C.  
     
     
         102 . The method according to  claim 2 , wherein the preparation of one or more digestive enzymes has a residual solvent content.  
     
     
         103 . The method according to  claim 102 , further comprising reducing said residual solvent content of the preparation of one or more digestive enzymes prior to said irradiation step.  
     
     
         104 . The method according to  claim 103 , wherein said reduced residual solvent content is less than about 10%.  
     
     
         105 . The method according to  claim 103 , wherein said reduced residual solvent content is less than about 1%.  
     
     
         106 . The method according to  claim 103 , wherein said reduced residual solvent content is less than about 0.2%.