Enabling the controlled assembly of antibody conjugates with a loading of two modules without antibody engineering

Abstract

A novel reagent/strategy enables the controlled assembly of antibody conjugates with a loading of two modules without antibody engineering.

Bioconjugation with Maleimides: A Useful Tool for Chemical Biology

“…The DBPD platform has been considerably explored in the generation of ADCs, antibody conjugates, antibody directed photosensitizers, protein‐protein conjugates and targeted nanotherapeutics . A DBPD reagent incorporating a terminal alkyne and cyclooctyne probe was used to accomplish complete rebridging of Trastuzumab (4:1 ratio of PD‐to‐antibody).…”

“…These advances paved the way to new applications in imaging, theranostics and new dual warhead antibiotics (Scheme A) . More recently DBPD technology has been applied in the production of more complex assemblies of antibody conjugates via controlled loading of two modules (via click handles) on a native antibody scaffold . This strategy provided a chemical solution to a challenge that had been only tackled using antibody engineering.…”

“…Brentuximab vedotin and trastuzumab emtansine, which target CD30 and HER2 + ,r espectively, [1] are two pioneering examples of second generation ADCs whose construction rely on maleimide chemistry.O nt heset herapeutics the cysteinem odification occurs rapidlya nd almostq uantitatively under physiological conditions, allowing the incorporation of higherl evels of cytotoxicd rug, and to some extentt ackling the heterogeneity problems associated with less efficient bioconjugation strategies. [19][20][21][22][23][24][25][26][27] However,t hiosuccinimide adducts are now known to undergo retro-Michael additions and/oro ther thiol exchange reactions under physiological conditions. Pharmacologically,t he consequences of premature thiosuccinimide cleavage fromt he ADCs may encompass decreasedt argeted cytotoxic activity as well as off-target toxicity.T hese concerns have inspired the development of novel maleimide reagents attempting to overcome these shortcomings,e ither by attenuating thiol exchange reactions (e.g.,via thiosuccinimide hydrolysis and tunable maleimide constructs) or by the development alternative chemotypestog enerate physiologically stable bioconjugates.…”

Characterization of Disulfide Bond Rebridged Fab–Drug Conjugates Prepared Using a Dual Maleimide Pyrrolobenzodiazepine Cytotoxic Payload

“…Rebridging of the native interchain disulfide bond has been used as a strategy to prepare Fab–drug conjugates (FDCs) and ADCs . For example, Godwin and colleagues developed a rebridging conjugation approach using a bis‐sulfone linker–drug, in which interchain disulfide bonds in either full‐length antibodies or Fab fragments were first partially reduced, followed by bis‐alkylation to conjugate both thiols of the two native cysteines.…”

“…The resulting ADCs were highly stable and had potent cytotoxicity against tumor cells. Caddick and colleagues contributed significantly over the past few years to the development of several rebridging approaches to prepare well‐defined FDCs and ADCs . Finally, rebridging of disulfide bonds through a thiol–yne reaction with a cyclooctyne has also been successfully applied to an antibody Fab …”