Multiple-enzyme nanocomplexes

Patent metadata

Field	Value
Publication number	US-10016490-B2
Application number	US-201214130873-A
Country	US
Kind code	B2
Filing date	Jul 6, 2012
Priority date	Jul 6, 2011
Publication date	Jul 10, 2018
Grant date	Jul 10, 2018

Abstract

Provided are nanocomplexes having at least two different enzymes and a polymeric network anchored to at least one of the enzymes. In some embodiments, the activities of the enzymes catalyze a cascade reaction.

First claim

The invention claimed is:

1. A multiple-enzyme nanocomplex comprising at least two different enzymes and a polymeric network which is anchored to at least one of said at least two different enzymes, wherein the multiple-enzyme nanocomplex has material properties that result from being prepared by a method comprising:
(a) selecting a first enzyme and a second enzyme for encapsulation;
(b) linking the first enzyme and the second enzyme to one another, wherein the linking comprises conjugating said enzymes to single nucleic acid strands that are hybridizable to each other and allowing the strands to hybridize;
(c) acrylating the linked enzymes produced by (b);
(d) combining the acrylated multiple-enzyme complex produced by (c) with polymerizable monomers so that the monomers are linked to the acrylated multiple-enzyme complex; and
(e) polymerizing the polymerizable monomers and the acrylated multiple-enzyme complex in situ on the surface of the multiple-enzyme complex so as to produce a polymeric network that forms a permeable shell that encapsulates the multiple-enzyme complex.

2. The multiple-enzyme nanocomplex according to claim 1 , wherein said polymeric network is anchored to all of said at least two different enzymes.

3. The multiple-enzyme nanocomplex according to claim 1 , wherein the activities of said two different enzymes catalyze a cascade reaction.

4. The multiple-enzyme nanocomplex according to claim 1 , wherein said at least two different enzymes are covalently or non-covalently linked to one another.

5. The multiple-enzyme nanocomplex according to claim 4 , wherein the covalent linkage is degradable in vivo.

6. The multiple-enzyme nanocomplex according to claim 1 , wherein said polymeric network comprises a polymer composed of at least one monomeric unit.

7. The multiple-enzyme nanocomplex according to claim 6 , wherein said polymeric network comprises a polymer composed of at least two different monomeric units.

8. The multiple-enzyme nanocomplex according to claim 1 , wherein the polymeric network encapsulates the multiple-enzyme nanocomplex in a manner sufficient to inhibit degradation of the multiple-enzyme nanocomplex when the multiple-enzyme nanocomplex is disposed in an in vivo environment.

9. The multiple-enzyme nanocomplex according to claim 8 , wherein the multiple-enzyme nanocomplex consists essentially of two different enzymes.

10. The multiple-enzyme nanocomplex according to claim 9 , wherein:
the first enzyme generates a first product in a first enzymatic reaction with a first substrate;
the second enzyme reacts with the first product in a second enzymatic reaction; and
the polymeric network exhibits a permeability sufficient to allow the first substrate to diffuse from an external environment outside of the shell to the first enzyme so that the first product is generated; and
the polymeric network exhibits a permeability sufficient to allow the first product to diffuse away from the first enzyme and to the second enzyme so that the second enzymatic reaction occurs.

11. The multiple-enzyme nanocomplex according to claim 10 , wherein:
the first enzyme is an alcohol oxidase; and
the second enzyme is a catalase.

12. The multiple-enzyme nanocomplex according to claim 11 , wherein the multiple-enzyme nanocomplex is combined with one or more filling agents, binding agents or buffering agents adapted for use in an orally administered formulation.

Assignees

• Lu Yunfeng
• Yan Ming
• Liu Yang
• Univ California

Inventors

• Lu Yunfeng
• Yan Ming
• Liu Yang

Classifications

• A61P39/02

  Antidotes · CPC title

• A61K47/64

  Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent (peptidic linkers A61K47/65) · CPC title

• A61K38/443 Primary

  acting on CH-OH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1) · CPC title

• B82Y5/00

  Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery · CPC title

• A61K38/44

  Oxidoreductases (1) · CPC title