Self-Immolative Carbamate Linkers for CD19-Budesonide Antibody-Drug Conjugates

Antibody-drug conjugates consist of potent small molecule payloads linked to a targeting antibody. Payloads must possess a viable functional group by which a linker for conjugation can be attached. Linker-attachment options remain limited for connection to payloads via hydroxyl groups. A releasing group based on 2-aminopyridine was developed to enable stable attachment of para-aminobenzyl carbamate (PABC) linkers to the C21-hydroxyl group of budesonide, a glucocorticoid receptor agonist. Payload release involves a cascade of two self-immolative events that are initiated by protease-mediated cleavage of the dipeptide-PABC bond. Budesonide release rates were determined for a series of payload-linker intermediates in buffered solution at pH 7.4 and pH 5.4, leading to the identification of 2-aminopyridine as the preferred releasing group. Addition of a polyethylene glycol group improved linker hydrophilicity, thereby providing CD19-budesonide ADCs with suitable properties. ADC23 demonstrated targeted budesonide delivery to CD19 expressing cells and inhibited B-cell activation in mice.