Site-Specific Tandem Knoevenagel Condensation-Michael Addition To Generate Antibody-Drug Conjugates
Site-Specific Tandem Knoevenagel Condensation-Michael Addition To Generate Antibody-Drug Conjugates
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DOI：
10.1021/acsmedchemlett.6b00253
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发表时间：
2016-11-01
影响因子：
4.2
通讯作者：
Rabuka, David
中科院分区：
文献类型：
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作者：
Kudirka, Romas A.;Barfield, Robyn M.;Rabuka, David
扩展连接技术是迫切需要的，以产生独特的连接功能增强蛋白质的不断增长的领域。为了解决这一需求，我们提出了一种独特的化学连接，涉及双添加的吡唑啉酮部分与标记的蛋白质。这种连接通过串联Knoevenagel缩合Michael加成发生。吡唑啉酮与醛反应生成烯酮，该烯酮随后受到第二吡唑啉酮的攻击，生成双吡唑啉酮物质。这种快速简便的连接技术是在不存在催化剂的温和条件下进行的，以产生以前通过常规连接反应无法获得的新结构。使用这种独特的连接，我们产生了三种位点特异性标记的抗体药物缀合物（ADC），平均四种药物对应一种抗体。报告了位点特异性ADC的体外和体内功效沿着以及药代动力学数据。
Expanded ligation techniques are sorely needed to generate unique linkages for the growing field of functionally enhanced proteins. To address this need, we present a unique chemical ligation that involves the double addition of a pyrazolone moiety with an aldehyde-labeled protein. This ligation occurs via a tandem Knoevenagel condensation Michael addition. A pyrazolone reacts with an aldehyde to generate an enone, which undergoes subsequent attack by a second pyrazolone to generate a bis-pyrazolone species. This rapid and facile ligation technique is performed under mild conditions in the absence of catalyst to generate new architectures that were previously inaccessible via conventional ligation reactions. Using this unique ligation, we generated three site-specifically labeled antibody drug conjugates (ADCs) with an average of four drugs to one antibody. The in vitro and in vivo efficacies along with pharmacokinetic data of the site-specific ADCs are reported.