Meijler Michael M, Kaufmann Gunnar F, Qi Longwu, Mee Jenny M, Coyle Avery R, Moss Jason A, Wirsching Peter, Matsushita Masayuki, Janda Kim D
Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, BCC-582, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
J Am Chem Soc. 2005 Mar 2;127(8):2477-84. doi: 10.1021/ja043935e.
Cocaine is a highly addictive drug, and despite intensive efforts, effective therapies for cocaine craving and addiction remain elusive. In recent years, we and others have reported advances in anti-cocaine immunopharmacotherapy based on specific antibodies capable of sequestering the drug before it reaches the brain. In an effort to obtain high affinity therapeutic anti-cocaine antibodies, either whole IgGs or other antibody constructs, fluorescence spectroscopic techniques could provide a means of assisting selection and engineering strategies. We report the synthesis of a series of cocaine-fluorophore conjugates (GNC-F1, GNC-F2, GNC-I) and the functional evaluation of these compounds against single-chain Fv antibodies obtained via crystallographic analysis/engineering and against commercially available anti-cocaine monoclonal antibodies with a wide range of cocaine-binding affinities. From these studies, we determined that the GNC-F2 fluorophore reproduced affinity constants obtained using [(3)H]-labeled cocaine. We anticipate that the readily synthesized and nonradioactive GNC-F2 will find use both as a tool for bioimaging and in the high-throughput selection and engineering of potential therapeutic antibodies against cocaine.
可卡因是一种极易上瘾的毒品，尽管人们付出了巨大努力，但针对可卡因成瘾和渴望的有效治疗方法仍然难以找到。近年来，我们和其他研究人员报告了基于特定抗体的抗可卡因免疫药物疗法取得的进展，这些抗体能够在药物进入大脑之前将其隔离。为了获得高亲和力的治疗性抗可卡因抗体（无论是完整的免疫球蛋白还是其他抗体构建体），荧光光谱技术可以提供一种辅助选择和工程策略的方法。我们报告了一系列可卡因-荧光团缀合物（GNC-F1、GNC-F2、GNC-I）的合成，以及这些化合物针对通过晶体学分析/工程获得的单链Fv抗体和针对具有广泛可卡因结合亲和力的市售抗可卡因单克隆抗体的功能评估。通过这些研究，我们确定GNC-F2荧光团重现了使用[³H]标记可卡因获得的亲和常数。我们预计，易于合成且无放射性的GNC-F2将既用作生物成像工具，又用于针对可卡因的潜在治疗性抗体的高通量筛选和工程设计。
