Cytotoxic
pyrrolobenzodiazepine (PBD)-dimer molecules are frequently
utilized as payloads for antibody–drug conjugates (ADCs), and
many examples are currently in clinical development. In order to further
explore this ADC payload class, the physicochemical properties of
various PBD-dimer molecules were modified by the systematic introduction
of acidic and basic moieties into their chemical structures. The impact
of these changes on DNA binding, cell membrane permeability, and in vitro antiproliferation potency was, respectively, determined
using a DNA alkylation assay, PAMPA assessments, and cell-based cytotoxicity
measurements conducted with a variety of cancer lines. The modified
PBD-dimer compounds were subsequently incorporated into CD22-targeting
ADCs, and these entities were profiled in a variety of in
vitro and in vivo experiments. The introduction
of a strongly basic moiety into the PBD-dimer scaffold afforded a
conjugate with dramatically worsened mouse tolerability properties
relative to ADCs derived from related payloads, which lacked the basic
group.