Mutant organophosphorus acid anhydrolases and uses thereof
Inventors
Guelta, Mark A • Dixon, Melissa M. • Harvey, Steven P
Assignees
United States Department of the Army • Government of the United States of America
Publication Number
US-10238904-B1
Publication Date
2019-03-26
Expiration Date
2037-09-13
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Abstract
Disclosed herein are non-wild-type organophosphorus acid anhydrolases having two site mutations, methods of production, and methods of use to effectively degrade toxic chemicals such as ((RS)-Propan-2-yl methylphosphonofluoridate) (Sarin) and other organophosphorus compounds.
Core Innovation
The invention provides non-wild-type organophosphorus acid anhydrolases (OPAAs) comprising two site mutations at sequence positions 212 and 342 of SEQ ID NO: 1. Specifically, the wild-type Tyrosine (Y) at position 212 is replaced with Phenylalanine (F), and the wild-type Valine (V) at position 342 is replaced with Isoleucine (I). These mutated OPAAs exhibit at least eight times greater catalytic efficiency for degrading the nerve agent Sarin ((RS)-Propan-2-yl methylphosphonofluoridate) compared to the wild-type enzyme. The invention also includes methods of producing these mutant enzymes, their use to degrade Sarin and other organophosphorus compounds, and kits containing the mutant OPAA enzyme with pharmaceutically acceptable carriers.
The problem addressed is that certain organophosphorus compounds, such as pesticides and chemical warfare nerve agents like Sarin, are highly toxic and hazardous to human health and the environment. While native OPAA enzymes can hydrolyze various organophosphorus compounds, their activity against Sarin is marginal and insufficient for practical detoxification or medical countermeasures. Existing mutant OPAAs with a single mutation at position 212 exhibit enhanced activity for some V-agents but not improved degradation of Sarin or other G-type nerve agents. Therefore, new compounds and methods that effectively detoxify Sarin and G-type nerve agents are needed.
Claims Coverage
The patent presents three main inventive features based on the independent claims relating to a mutant OPAA enzyme, a method for degrading Sarin, and a kit for degradation involving the mutant enzyme.
Enzyme with dual mutations at positions 212 and 342
An isolated OPAA enzyme with non-wild-type amino acids at sequence positions 212 and 342 of SEQ ID NO: 1, wherein Tyrosine (Y) at position 212 is replaced by Phenylalanine (F) and Valine (V) at position 342 is replaced by Isoleucine (I).
Method of degrading Sarin using the mutant enzyme
A method for degrading ((RS)-Propan-2-yl methylphosphonofluoridate) (Sarin) by contacting it with the isolated OPAA enzyme bearing the dual mutations at positions 212 and 342 as described.
Kit comprising the mutant enzyme and pharmaceutical carrier
A kit for degrading Sarin comprising the isolated OPAA enzyme having the mutations at positions 212 and 342, along with a pharmaceutically acceptable carrier and optionally other adjuvants or excipients.
These inventive features cover the mutated OPAA enzyme with dual amino acid substitutions, its direct application in Sarin degradation, pharmaceutical formulations for treatment, and kits including the enzyme for detoxification purposes.
Stated Advantages
The engineered non-wild-type OPAA has at least eight times greater catalytic efficiency for degrading Sarin compared to wild-type enzyme.
The mutant enzyme enables effective catalytic degradation of highly toxic chemical warfare agents like Sarin, improving potential use as decontaminants or medical countermeasures.
Documented Applications
In vivo treatment of Sarin poisoning by administering pharmaceutical compositions containing the mutant OPAA.
Catalytic decontamination of Sarin-contaminated surfaces or environments via application of mutant OPAA enzyme.
Uses in pharmaceutical formulations administered by various routes including intravenous, subcutaneous, intraperitoneal, oral, mucosal, and inhalational delivery.
Kits for degrading Sarin containing the mutant enzyme and pharmaceutical carriers.
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