Oral delivery of enzymes by nanocapsules for targeted metabolism of alcohol or toxic metabolites

Abstract

The invention disclosed herein includes nanocomplexes that are designed include enzymes that have complementary functional attributes and methods for using these nanocomplexes. Illustrative examples include nanocomplexes that comprise both an alcohol oxidase enzyme as well as a catalase enzyme. These nanocomplexes can be used in methods designed to lower blood alcohol levels in vivo, and/or to break down the toxic byproducts of alcohol metabolism. Consequently these nanocomplexes can be used to treat a variety of conditions resulting from the consumption of alcohol, including for example, acute alcohol intoxication.

1 . A method of decreasing a concentration of hydrogen peroxide in an aqueous environment comprising the steps of:
 (a) introducing a multiple-enzyme nanocomplex system into the aqueous environment, wherein the multiple-enzyme nanocomplex system comprises:
 a first enzyme that generates hydrogen peroxide in a first enzymatic reaction with a first substrate present in the aqueous environment; 
 a second enzyme that converts the hydrogen peroxide into water in a second enzymatic reaction; and 
 a polymeric network configured to form a shell that encapsulates the first enzyme and the second enzyme, wherein: 
 the polymeric network encapsulates the first enzyme and the second enzyme in a manner that inhibits degradation of the first enzyme and the second enzyme when the multiple-enzyme nanocomplex is disposed in an in vivo environment; 
 the polymeric network exhibits a permeability sufficient to allow the substrate to diffuse from an external environment outside of the shell to the first enzyme so that the hydrogen peroxide is generated; 
 the polymeric network exhibits a permeability sufficient to allow the hydrogen peroxide to diffuse away from the first enzyme and to the second enzyme so that the water is generated; and 
 the first enzyme is coupled to the second enzyme; or 
 the polymeric network is coupled to the first enzyme or the second enzyme; and 
   (b) allowing the first enzyme and the second enzyme in the multiple-enzyme nanocomplex system to react with the substrate and generate hydrogen peroxide and water;   so that the concentration of hydrogen peroxide in the aqueous environment is decreased.   
     
     
         2 . The method of  claim 1 , wherein at least one enzyme encapsulated within the polymeric network is selected from the group consisting of an alcohol oxidase, a catalase, a peroxidase, an alcohol dehydrogenase, an aldehyde dehydrogenase or an acetaldehyde oxidase. 
     
     
         3 . The method of  claim 2 , wherein the first enzyme is an alcohol oxidase. 
     
     
         4 . The method of  claim 3 , wherein the second enzyme is a catalase. 
     
     
         5 . The method of  claim 4 , wherein the aqueous environment comprises an in vivo environment. 
     
     
         6 . The method of  claim 1 , wherein:
 the first enzyme is a glucose oxidase; and   the second enzyme is a horseradish peroxidase.   
     
     
         7 . The method of  claim 6 , wherein the aqueous environment comprises an in vitro environment. 
     
     
         8 . The method of  claim 2 , wherein the multiple-enzyme nanocomplex system reduces blood ethanol concentrations in the individual by at least 25, 50, 75 or 100 mg/dL within 90 minutes following administration to the individual. 
     
     
         9 . The method of  claim 2 , further comprising a third enzyme encapsulated within the polymeric network, wherein the third enzyme is an alcohol dehydrogenase, an aldehyde dehydrogenase or an acetaldehyde oxidase. 
     
     
         10 . The method of  claim 1 , wherein the first enzyme and the second enzyme are coupled together by strands of complimentary polynucleotides. 
     
     
         11 . A method of decreasing a concentration of hydrogen peroxide in an aqueous environment comprising the steps of:
 (a) introducing a multiple-enzyme nanocomplex system into the aqueous environment, wherein the multiple-enzyme nanocomplex system comprises:
 a first enzyme that generates hydrogen peroxide in a first enzymatic reaction with a first substrate present in the aqueous environment; 
 a second enzyme that converts the hydrogen peroxide into water in a second enzymatic reaction; and 
 a polymeric network configured to form a shell that encapsulates the first enzyme and the second enzyme, wherein: 
 the polymeric network encapsulates the first enzyme and the second enzyme in a manner that inhibits degradation of the first enzyme and the second enzyme when the multiple-enzyme nanocomplex is disposed in an in vivo environment; 
 the polymeric network exhibits a permeability sufficient to allow the substrate to diffuse from an external environment outside of the shell to the first enzyme so that the hydrogen peroxide is generated; 
 the first enzyme is coupled to the second enzyme; or 
 the polymeric network is coupled to the first enzyme or the second enzyme; and 
 the polymeric shell is formed over the coupled first enzyme or the coupled second enzyme so as to dispose the first enzyme and the second enzyme in a proximal orientation selected to facilitate hydrogen peroxide produced in the first enzymatic reaction diffusing to the second enzyme; and 
   (b) allowing the first enzyme and the second enzyme in the multiple-enzyme nanocomplex system to react with the substrate and generate hydrogen peroxide and water;   so that the concentration of hydrogen peroxide in the aqueous environment is decreased.   
     
     
         12 . The method of  claim 11 , wherein at least one enzyme encapsulated within the polymeric network is selected from the group consisting of an alcohol oxidase, a catalase, a peroxidase, an alcohol dehydrogenase, an aldehyde dehydrogenase or an acetaldehyde oxidase. 
     
     
         13 . The method of  claim 12 , wherein:
 the first enzyme is an alcohol oxidase; and   the second enzyme is a catalase.   
     
     
         14 . The method of  claim 13 , wherein the aqueous environment comprises an in vivo environment. 
     
     
         15 . The method of  claim 13 , further comprising a third enzyme encapsulated within the polymeric network, wherein the third enzyme is an alcohol dehydrogenase, an aldehyde dehydrogenase or an acetaldehyde oxidase.