Improved pharmacokinetics and immunogenicity profile of organophosphorus hydrolase by chemical modification with polyethylene glycol
详细信息 查看全文
- 作者：Boris N. Novikov ; Janet K. Grimsley ; Rory J. Kern ; James R. Wild ; Melinda E. Wales
- 关键词：Organophosphorus hydrolase ; PEG ; PEGylation ; Pharmacokinetics ; Immunogenicity
- 刊名：Journal of Controlled Release
- 出版年：2010
- 出版时间：15 September 2010
- 年：2010
- 卷：146
- 期：3
- 页码：318-325
- 全文大小：481 K
文摘
A catalytic bioscavenger with broad substrate specificity for the therapeutic and prophylactic defense against recognized chemical threat agents has been a long standing objective of civilian and military research. A catalytic bioscavenger utilizing the bacterial enzyme organophosphorus hydrolase (OPH) is characterized in these studies, and has potential application for both military and civilian personnel in threat scenarios involving either nerve agents or OP pesticides. The present study examines the effects of PEGylation on the biochemical and pharmacological characteristics of OPH. The enzyme was conjugated with linear and branched methyl-PEOn-NHS esters of relatively small molecular mass from 333 to 2420 Da. PEGylated OPH displayed a decreased maximal catalytic rate, though substantial activity was maintained against two tested substrates: up to 30 % with paraoxon and up to 50–60 % with demeton-S. The thermostability of the PEGylated enzymes ranged between 60 and 64 °C, compared to the unmodified OPH, which is approximately 67 °C. The enzyme conjugates revealed a significant improvement of pharmacokinetic properties in animal studies. The clearance from a guinea pig's blood stream significantly decreased relative to unmodified OPH, resulting in an increase of residence time and systemic availability. Evaluation of the humoral immune response indicated that the branched PEG–OPH conjugate significantly reduced production of anti-OPH antibodies, compared to the unmodified enzyme. The OPH–PEG conjugates with improved pharmacokinetic and immunogenicity properties, considerable catalytic activity and thermal stability provide a new opportunity for the in vivo detoxification of the neurotoxic OP compounds.