Bridging Disulfides for Stable and Defined Antibody Drug Conjugates

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• 作者：George Badescu ; Penny Bryant ; Matthew Bird ; Korinna Henseleit ; Julia Swierkosz ; Vimal Parekh ; Rita Tommasi ; Estera Pawlisz ; Kosma Jurlewicz ; Monika Farys ; Nicolas Camper ; XiaoBo Sheng ; Martin Fisher ; Ruslan Grygorash ; Andrew Kyle ; Amrita Abhilash ; Mark Frigerio ; Jeff Edwards ; Antony Godwin
• 刊名：Bioconjugate Chemistry
• 出版年：2014
• 出版时间：June 18, 2014
• 年：2014
• 卷：25
• 期：6
• 页码：1124-1136
• 全文大小：539K
• 年卷期：v.25,no.6(June 18, 2014)
• ISSN：1520-4812

文摘

To improve both the homogeneity and the stability of ADCs, we have developed site-specific drug-conjugating reagents that covalently rebridge reduced disulfide bonds. The new reagents comprise a drug, a linker, and a bis-reactive conjugating moiety that is capable of undergoing reaction with both sulfur atoms derived from a reduced disulfide bond in antibodies and antibody fragments. A disulfide rebridging reagent comprising monomethyl auristatin E (MMAE) was prepared and conjugated to trastuzumab (TRA). A 78% conversion of antibody to ADC with a drug to antibody ratio (DAR) of 4 was achieved with no unconjugated antibody remaining. The MMAE rebridging reagent was also conjugated to the interchain disulfide of a Fab derived from proteolytic digestion of TRA, to give a homogeneous single drug conjugated product. The resulting conjugates retained antigen-binding, were stable in serum, and demonstrated potent and antigen-selective cell killing in in vitro and in vivo cancer models. Disulfide rebridging conjugation is a general approach to prepare stable ADCs, which does not require the antibody to be recombinantly re-engineered for site-specific conjugation.