Anti-HER2, high-DAR Antibody Fragment Drug Conjugates (FDC) with a Glucuronide-based MMAE linker-payload demonstrates superior efficacy over IgG-based ADCs

Abstract Antibody-Drug Conjugates (ADCs) are poised to embed themselves as pillars of cancer therapy after decades of development and fine-tuning. The vast majority of those in preclinical and clinical development are based on full-length immunoglobulins employing a variety of linker-payloads and conjugation strategies. Other, smaller formats are being considered to overcome some of the current limitations of ADCs, notably poor solid tumour penetration and prolonged systemic toxin exposure. By combining stable, high-lysine containing single-chain Fv antibody fragments, compact hydrophilic linkers and a validated MMAE (monomethyl auristatin-E) payload, high-DAR (Drug:Antibody Ratio) FDCs (Antibody-Fragment Drug Conjugates) were made which retained significant binding and developability properties. Against the established target HER2, an average DAR 6 FDC was reproducibly obtained (equivalent to a DAR 30 ADC by mass) with picomolar binding affinity, low aggregation and translatable pharmacokinetics. Despite the faster elimination kinetics, rapid and intense tumour payload delivery was seen leading to tumour cure efficacy in multiple HER2 tumour xenografts at doses as low as 0.6mg/kg given 4-times, weekly. Internalization and tumour uptake quantification data illustrate the benefits of the higher-penetrating format. Experience with over a dozen linker-payload structures has provided an insight into the critical design features that could make FDCs a viable alternative to ADCs in the most challenging solid tumour indications.