title: Orthogonal Cysteine Protection Enables Homogeneous Multi-Drug Antibody-Drug Conjugates.
abstract: A strategy for the preparation of homogeneous antibody-drug conjugates (ADCs) containing multiple payloads has been developed. This approach utilizes sequential unmasking of cysteine residues with orthogonal protection to enable site-specific conjugation of each drug. In addition, because the approach utilizes conjugation to native antibody cysteine residues, it is widely applicable and enables high drug loading for improved ADC potency. To highlight the benefits of ADC dual drug delivery, this strategy was applied to the preparation of ADCs containing two classes of auristatin drug-linkers that have differing physiochemical properties and exert complementary anti-cancer activities. Dual-auristatin ADCs imparted activity in cell line and xenograft models that are refractory to ADCs comprised of the individual auristatin components. This work presents a facile method for construction of potent dual-drug ADCs and demonstrates how delivery of multiple cytotoxic warheads can lead to improved ADC activities. Lastly, we anticipate that the conditions utilized herein for orthogonal cysteine unmasking are not restricted to ADCs and can be broadly utilized for site-specific protein modification.
authors.affiliation: Seattle Genetics, Inc., 21823 30thDrive SE, Bothell, WA, USA.
authors.forename: Matthew R
authors.identifier: 0000-0002-3762-9983
authors.initials: MR
authors.lastname: Levengood
authors.affiliation: Seattle Genetics, Inc., 21823 30thDrive SE, Bothell, WA, USA.
authors.forename: Xinqun
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Seattle Genetics, Inc., 21823 30thDrive SE, Bothell, WA, USA.
authors.forename: Joshua H
authors.initials: JH
authors.lastname: Hunter
authors.affiliation: Seattle Genetics, Inc., 21823 30thDrive SE, Bothell, WA, USA.
authors.forename: Kim K
authors.initials: KK
authors.lastname: Emmerton
authors.affiliation: Seattle Genetics, Inc., 21823 30thDrive SE, Bothell, WA, USA.
authors.forename: Jamie B
authors.initials: JB
authors.lastname: Miyamoto
authors.affiliation: Seattle Genetics, Inc., 21823 30thDrive SE, Bothell, WA, USA.
authors.forename: Timothy S
authors.initials: TS
authors.lastname: Lewis
authors.affiliation: Seattle Genetics, Inc., 21823 30thDrive SE, Bothell, WA, USA.
authors.forename: Peter D
authors.initials: PD
authors.lastname: Senter
mesh_terms: D062365:Aminobenzoates / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000970:Antineoplastic Agents / Q000737:chemistry*; D003545:Cysteine / Q000737:chemistry*; D016503:Drug Delivery Systems; D018796:Immunoconjugates / Q000737:chemistry*; D008968:Molecular Conformation; D009842:Oligopeptides / Q000737:chemistry*
chemical_list: D062365:Aminobenzoates; D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D018796:Immunoconjugates; D009842:Oligopeptides; C543533:auristatin; D003545:Cysteine
keywords: antibodies; bioconjugate; cancer; cysteine; drug delivery