Development of Fc-Fused Cocaine Hydrolase for Cocaine Addiction Treatment: Catalytic and Pharmacokinetic Properties
- 271 Downloads
Cocaine abuse is a worldwide public health and social problem without a US Food and Drug Administration (FDA)-approved medication. Accelerating cocaine metabolism that produces biologically inactive metabolites by administration of an efficient cocaine hydrolase (CocH) has been recognized as a promising strategy for cocaine abuse treatment. However, the therapeutic effects of CocH are limited by its short biological half-life (e.g., 8 h or shorter in rats). In this study, we designed and prepared a set of Fc-fusion proteins constructed by fusing Fc(M3) with CocH3 at the N-terminus of CocH3. A linker between the two protein domains was optimized to improve both the biological half-life and catalytic activity against cocaine. It has been concluded that Fc(M3)-G6S-CocH3 not only has fully retained the catalytic efficiency of CocH3 against cocaine but also has the longest biological half-life (e.g., ∼ 136 h in rats) among all of the long-acting CocHs identified so far. A single dose (0.2 mg/kg, IV) of Fc(M3)-G6S-CocH3 was able to significantly attenuate 15 mg/kg cocaine-induced hyperactivity for at least 11 days (268 h) after the Fc(M3)-G6S-CocH3 administration.
KEY WORDScocaine drug abuse metabolic enzyme protein engineering
This work was supported in part by the National Institutes of Health (NIH grants UH2/UH3 DA041115, R01 DA035552, R01 DA032910, R01 DA013930, and R01 DA025100) and the National Science Foundation (NSF grant CHE-1111761).
Compliance with Ethical Standards
Experiments were performed in a same colony room in accordance with the Guide for the Care and Use of Laboratory Animals as adopted and promulgated by the National Institutes of Health. The animal protocol was approved by the IACUC (Institutional Animal Care and Use Committee) at the University of Kentucky.
The authors declare that they have no conflict of interest.
- 1.UNODC. World drug report 2014. New York: United Nations Publications; 2014.Google Scholar
- 2.Schrank KS. Cocaine-related emergency department presentations. NIDA Res Monogr. 1993;123:110–28.Google Scholar
- 3.Brim RL, Noon KR, Collins GT, Nichols J, Narasimhan D, Sunahara RK, et al. The ability of bacterial cocaine esterase to hydrolyze cocaine metabolites and their simultaneous quantification using high-performance liquid chromatography-tandem mass spectrometry. Mol Pharmacol. 2011;80(6):1119–27. https://doi.org/10.1124/mol.111.074534.CrossRefPubMedPubMedCentralGoogle Scholar
- 4.Abuse S, Administration MHS. The DAWN report: highlights of the 2011 drug abuse warning network (DAWN) findings on drug-related emergency department visits. Rockville: SAMHSA; 2012.Google Scholar
- 15.Collins GT, Narasimhan D, Cunningham AR, Zaks ME, Nichols J, Ko M-C, et al. Long-lasting effects of a PEGylated mutant cocaine esterase (CocE) on the reinforcing and discriminative stimulus effects of cocaine in rats. Neuropsychopharmacology. 2012;37(5):1092–103. https://doi.org/10.1038/npp.2011.226.CrossRefPubMedGoogle Scholar
- 16.Brimijoin S, Gao Y, Anker JJ, Gliddon LA, LaFleur D, Shah R, et al. A cocaine hydrolase engineered from human butyrylcholinesterase selectively blocks cocaine toxicity and reinstatement of drug seeking in rats. Neuropsychopharmacology. 2008;33(11):2715–25. https://doi.org/10.1038/sj.npp.1301666.CrossRefPubMedPubMedCentralGoogle Scholar
- 18.Cohen-Barak O, Wildeman J, van de Wetering J, Hettinga J, Schuilenga-Hut P, Gross A, et al. Safety, pharmacokinetics, and pharmacodynamics of TV-1380, a novel mutated Butyrylcholinesterase treatment for cocaine addiction, after single and multiple intramuscular injections in healthy subjects. J Clin Pharmacol. 2015;55:573–83. https://doi.org/10.1002/jcph.450.CrossRefPubMedPubMedCentralGoogle Scholar
- 19.Shram MJ, Cohen-Barak O, Chakraborty B, Bassan M, Schoedel KA, Hallak H, et al. Assessment of pharmacokinetic and pharmacodynamic interactions between albumin-fused mutated butyrylcholinesterase and intravenously administered cocaine in recreational cocaine users. J Clin Psychopharmacol. 2015;35:396–405. https://doi.org/10.1097/JCP.0000000000000347.CrossRefPubMedGoogle Scholar
- 23.Xue L, Ko M-C, Tong M, Yang W, Hou S, Fang L, et al. Design, preparation, and characterization of high-activity mutants of human butyrylcholinesterase specific for detoxification of cocaine. Mol Pharmacol. 2011;79:290–7. https://doi.org/10.1124/mol.110.068494.CrossRefPubMedPubMedCentralGoogle Scholar
- 34.Schneider JD, Marillonnet S, Castilho A, Gruber C, Werner S, Mach L, et al. Oligomerization status influences subcellular deposition and glycosylation of recombinant butyrylcholinesterase in Nicotiana benthamiana. Plant Biotechnol J. 2014;12(7):832–9. https://doi.org/10.1111/pbi.12184.CrossRefPubMedPubMedCentralGoogle Scholar