Cocaine abuse is a major medical and public health problem. There is still no FDA-approved anti-cocaine medication. Disastrous medical and social consequences of cocaine abuse have made the development of an anti-cocaine medication a high priority. Enhancing cocaine metabolism by administration of human butyrylcholinesterase (BChE) is recognized as an efficient treatment strategy for cocaine overdose and addiction. However, the catalytic efficiency (kcat/KM) of wild-type BChE against the naturally occurring (-)- cocaine is low (kcat = 4.1 min-1 and KM = 4.5 ?M). Nevertheless, we have recently designed and discovered a set of BChE mutants, known as cocaine hydrolases (CocHs), with >1,000-fold improved catalytic efficiency against (-)-cocaine compared to wild-type BChE. In vivo evidences and clinical data for the first one of our discovered and patented CocHs have demonstrated that our discovered CocHs are promising candidates for development of an anti-cocaine medication. Our recently designed, discovered and patented CocHs are significantly more potent. Built on our success in rational design and discovery of the CocHs, the currently proposed investigation is focused on rational design, preparation, and preclinical testing of a novel type of long-lasting CocH entities, denoted as Fc-CocH, obtained from fusion of CocH with Fc portion of human antibody IgG1.
The specific aims are: (1) to design new molecular entities of Fc-CocH that potentially have not only a high catalytic efficiency against (-)-cocaine, but also a long biologicl half-life;(2) to prepare and test the designed Fc-CocHs for their in vitro activities;(3) to characterize the in vivo potency, pharmacokinetics, and immunogenicity of Fc-CocHs in rats and rhesus monkeys;(4) to evaluate the actual effects of the promising Fc-CocHs (identified in Aim 3) on the physiological and behavior responses of animals to cocaine by performing cardiovascular assays and self-administration assays in rhesus monkeys. Accomplishment of this proposed investigation will result in the identification and development of the best possible Fc-CocH entity that has not only a high in vivo potency in blocking physiological effects of cocaine, but also a long biological half- life without immunogenicity. The long-lasting Fc-CocH entity optimized in this investigation is expected to be highly effective and safe as a novel exogenous enzyme suitable for cocaine addiction treatment in humans.

Public Health Relevance

Accelerating cocaine metabolism has been recognized as a promising treatment strategy for cocaine overdose and abuse. The long-lasting cocaine-metabolizing enzymes to be tested in this project are promising candidates for an efficient anti-cocaine medication.

Agency
National Institute of Health (NIH)
Institute
National Institute on Drug Abuse (NIDA)
Type
Research Project (R01)
Project #
5R01DA035552-02
Application #
8636423
Study Section
Special Emphasis Panel (ZDA1-GXM-A (11))
Program Officer
Shih, Ming L
Project Start
2013-04-01
Project End
2016-02-29
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
2
Fiscal Year
2014
Total Cost
$1,097,704
Indirect Cost
$272,090
Name
University of Kentucky
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
939017877
City
Lexington
State
KY
Country
United States
Zip Code
40506
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Fang, Lei; Chow, K Martin; Hou, Shurong et al. (2014) Rational design, preparation, and characterization of a therapeutic enzyme mutant with improved stability and function for cocaine detoxification. ACS Chem Biol 9:1764-72
Hou, Shurong; Zhan, Max; Zheng, Xirong et al. (2014) Kinetic characterization of human butyrylcholinesterase mutants for the hydrolysis of cocaethylene. Biochem J 460:447-57
Zheng, Fang; Xue, Liu; Hou, Shurong et al. (2014) A highly efficient cocaine-detoxifying enzyme obtained by computational design. Nat Commun 5:3457
Qiao, Yan; Han, Keli; Zhan, Chang-Guo (2014) Reaction pathways and free energy profiles for cholinesterase-catalyzed hydrolysis of 6-monoacetylmorphine. Org Biomol Chem 12:2214-27
Fang, Lei; Hou, Shurong; Xue, Liu et al. (2014) Amino-acid mutations to extend the biological half-life of a therapeutically valuable mutant of human butyrylcholinesterase. Chem Biol Interact 214:18-25
Zheng, Fang; Zhan, Max; Huang, Xiaoqin et al. (2014) Modeling in vitro inhibition of butyrylcholinesterase using molecular docking, multi-linear regression and artificial neural network approaches. Bioorg Med Chem 22:538-49
Fang, Lei; Zheng, Fang; Zhan, Chang-Guo (2014) A model of glycosylated human butyrylcholinesterase. Mol Biosyst 10:348-54
Lu, Haiting; Huang, Xiaoqin; AbdulHameed, Mohamed Diwan M et al. (2014) Binding free energies for nicotine analogs inhibiting cytochrome P450 2A6 by a combined use of molecular dynamics simulations and QM/MM-PBSA calculations. Bioorg Med Chem 22:2149-56
Wei, Donghui; Fang, Lei; Tang, Mingsheng et al. (2013) Fundamental reaction pathway for peptide metabolism by proteasome: insights from first-principles quantum mechanical/molecular mechanical free energy calculations. J Phys Chem B 117:13418-34

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