# US 7,070,973 B2 · Butyrylcholinesterase variants and methods of use

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 7,070,973 B2
- Document ID: US7070973B2
- Status: Granted
- Kind: B2
- Application number: 09/748,739
- Filed: 2000-12-26
- Published: 2002-08-29
- Granted: 2006-07-04
- Earliest claimed: 2000-12-26
- Assignees: Board of Regents of the University of Nebraska; Applied Molecular Evolution, Inc.
- Inventors: Oksana Lockridge; Jeffry D. Watkins
- Web page: https://patentgrep.com/patent/US7070973B2

## CPC

- C12Q 1/46: involving cholinesterase
- C12N 9/18: Carboxylic ester hydrolases {(3.1.1)}
- C12Y 301/01008: Cholinesterase (3.1.1.8), i.e. butyrylcholine-esterase
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## Abstract

The invention provides four butyrylcholinesterase variants having increased cocaine hydrolysis activity as well as the corresponding encoding nucleic acids. The invention also provides libraries comprising butyrylcholinesterase variants having at least one amino acid alteration in one or more regions of butyrylcholinesterase and further having at least one butyrylcholinesterase variant exhibiting enhanced cocaine hydrolysis activity compared to butyrylcholinesterase. The invention further provides methods of hydrolyzing a cocaine-based butyrylcholinesterase substrate as well as methods of treating a cocaine-induced condition.

## Claims

### Claim 1

A butyrylcholinesterase variant comprising an amino acid sequence comprising a sequence of amino acid residues 1 or 29 through 602, inclusive, of the amino acid sequence shown as SEQ ID NO:2, wherein said variant has tryptophan at amino acid position 356.

### Claim 2 (depends on 1)

The butyrylcholinesterase variant of claim 1, having a 15-fold increase in cocaine hydrolysis activity compared to human butyrylcholinesterase.