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Significance A number of antibody–drug conjugates (ADCs) with varying linkers carrying an uncialamycin analogue as payload were synthesized and tested for their cytotoxicity in vitro and in PDX mouse models. Importantly, a number of these enediyne-containing ADCs were found to exhibit potent and selective in vivo and in vitro cytotoxicities and also displayed a significant bystander killing effect. The latter finding is of particular importance since the currently approved enediyne ADCs (featuring an N-acetylated calicheamicinγ 1 I derivative as payload) are known not to induce bystander killing, thus lacking a possibly beneficial characteristic that could potentially improve the efficacy of oncological therapies.

Uncialamycin-based antibody–drug conjugates: Unique enediyne ADCs exhibiting bystander killing effect