Polymer engineered regenerating bioscavengers

Patent metadata

Field	Value
Publication number	US-2016244741-A1
Application number	US-201415026093-A
Country	US
Kind code	A1
Filing date	Oct 3, 2014
Priority date	Oct 3, 2013
Publication date	Aug 25, 2016
Grant date	—

Abstract

Embodiments of the invention provide at least one polymer covalently conjugated to an esterase. The at least one polymer includes a plurality of oxime functional groups.

First claim

1 .- 69 . (canceled)

70 . A composition comprising:
at least one polymer covalently conjugated to an enzyme, the at least one polymer comprising a reactive functional group, wherein the reactive functional group reacts with an inhibitor attached to an active site of the enzyme.

71 . The composition of claim 70 , wherein the enzyme comprises an esterase, and the reactive functional group comprises a plurality of oxime functional groups.

72 . The composition of claim 71 wherein the esterase comprises a member selected from the group consisting of acetylcholinesterase and butyrylcholinesterase.

73 . The composition of claim 71 wherein the esterase comprises chymotrypsin.

74 . The composition of claim 71 wherein the plurality of oxime functional groups comprises alkyne derivatives of 2-pyridine aldoxime.

75 . The composition of claim 71 wherein the plurality of oxime functional groups comprises an aldoxime.

76 . The composition of claim 71 wherein the plurality of oxime functional groups comprises a ketoxime.

77 . The composition of claim 70 wherein the reactive functional group reacts with the inhibitor attached to the active site of the enzyme releasing the inhibitor from the active site and restoring activity to the active site of the enzyme.

78 . The composition of claim 70 wherein the at least one polymer comprises at least one environmentally responsive monomer.

79 . The composition of claim 70 wherein the at least one polymer is a co-polymer comprising at least two different monomers, wherein at least one monomer comprises a member selected from the group consisting of aldoximes, ketoximes, muco-adhesion monomers, polyethylene glycol, bis-pyridinium oximes, N,N-dimethylacrylamide, N-isopropylacrylamide, (meth)acrylate, N,N-dimethylaminoethyl methacrylate, carboxyl acrylamide, 2-hydroxylethylmethacrylate, N-(2-hydroxypropyl)methacrylamide, quaternary ammonium monomers, sulfobetain methacrylate, oligo(ethylene glycol)methyl ether methacrylate, 2-PAM monomers, 4-PAM monomers, Clickable azide monomers, and combinations thereof.

80 . A composition comprising:
a bioconjugate comprising: an enzyme; and at least one polymer covalently conjugated to the enzyme, wherein: the at least one polymer comprises a plurality of reactive functional groups; and at least one reactive functional group of the plurality of reactive functional groups is positioned to react, in use, with a functional group on an inhibitor attached to an active site of the enzyme.

81 . The composition of claim 80 , wherein the enzyme comprises an esterase, and the at least one reactive functional group comprises a plurality of oxime functional groups.

82 . The composition of claim 81 wherein the at least one reactive functional group comprises an oxime functional group positioned to exert a nucleophilic attack on a phosphoryl functional group when, in use, the inhibitor includes the phosphoryl functional group attached to the active site of the esterase, said nucleophilic attack resulting in removal of the phosphoryl functional group from the active site restoring activity to the esterase.

83 . The composition of claim 81 wherein the at least one polymer of the bioconjugate comprises a flexibility sufficient to react, in use, with a phosphoryl functional group when an inhibitor having the phosphoryl functional group attaches to the active site of the esterase.

84 . The composition of claim 81 wherein the at least one polymer covalently conjugated to the esterase forms a long lived covalent conjugate that is maintained in the body for a time period ranging from more than about one day to more than about one week.

85 . The composition of claim 81 wherein the at least one polymer is a co-polymer comprising at least two different monomers, wherein at least one monomer comprises a member selected from the group consisting of aldoximes, ketoximes, muco-adhesion monomers, polyethylene glycol, bis-pyridinium oximes, N,N-dimethylacrylamide, N-isopropylacrylamide, (meth)acrylate, N,N-dimethylaminoethyl methacrylate, carboxyl acrylamide, 2-hydroxylethylmethacrylate, N-(2-hydroxypropyl)methacrylamide, quaternary ammonium monomers, sulfobetain methacrylate, oligo(ethylene glycol)methyl ether methacrylate, 2-PAM monomers, 4-PAM monomers, Clickable azide monomers, and combinations thereof.

86 . A method comprising:
administering a bioscavenger to an individual suffering from organophosphate toxin exposure, the bioscavenger comprising at least one polymer covalently conjugated to an esterase, the at least one polymer comprising a plurality of oxime functional groups.

87 . The method of claim 86 wherein, upon exposure of the bioscavenger to an organophosphate toxin, the method further comprises reacting at least one of the plurality of oxime functional groups with at least one covalently inhibited residue of the esterase to detoxify the organophosphate toxin and regenerate the bioscavenger.

88 . The method of claim 86 wherein the plurality of oxime functional groups comprises an oxime functional group positioned to exert a nucleophilic attack on a phosphoryl functional group when, in use, an inhibitor having the phosphoryl functional group attaches to an active site of the esterase, said nucleophilic attack resulting in removal of the phosphoryl functional group from the active site.

89 . The method of claim 86 wherein the esterase comprises chymotrypsin and the plurality of oxime functional groups comprises an aldoxime.

Assignees

• Univ Carnegie Mellon

Inventors

• Russell Alan J
• Murata Hironobu
• Cummings Chad
• Koepsel Richard R

Classifications

• C12N9/96 Primary

  Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates · CPC title

• C12N9/6427

  Chymotrypsins (3.4.21.1; 3.4.21.2); Trypsin (3.4.21.4) · CPC title

• C12Y304/21001

  Chymotrypsin (3.4.21.1) · CPC title

• A61K38/465

  acting on ester bonds (3.1), e.g. lipases, ribonucleases · CPC title

• C12N9/16

  acting on ester bonds (3.1) · CPC title