American Association for Cancer Research
15357163mct160825-sup-175098_4_supp_4009287_xp4px8.docx (242.61 kB)

Supplementary Data from Preclinical Evaluation of MEDI0641, a Pyrrolobenzodiazepine-Conjugated Antibody–Drug Conjugate Targeting 5T4

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journal contribution
posted on 2023-04-03, 15:48 authored by Jay Harper, Christopher Lloyd, Nazzareno Dimasi, Dorin Toader, Rose Marwood, Leeanne Lewis, David Bannister, Jelena Jovanovic, Ryan Fleming, Francois D'Hooge, Shenlan Mao, Allison M. Marrero, Martin Korade, Patrick Strout, Linda Xu, Cui Chen, Leslie Wetzel, Shannon Breen, Lilian van Vlerken-Ysla, Sanjoo Jalla, Marlon Rebelatto, Haihong Zhong, Elaine M. Hurt, Mary Jane Hinrichs, Keven Huang, Philip W. Howard, David A. Tice, Robert E. Hollingsworth, Ronald Herbst, Adeela Kamal

Supplementary Figure S1. Comparable internalization rates for 5T4_0108 and the 5T4-Tub ADC;Supplementary Figure S2. Affinity optimization improves in vitro cytotoxicity of a 5T4-ADC;Supplementary Figure S3. Induction of apoptosis by 5T4-PBD and 5T4-Tub;Supplementary Figure S4. 5T4 is expressed on CSCs of multiple cancer cell lines;Supplementary Figure S5. Only PBD is capable of inducing cytotoxicity of CSC populations in vitro;Supplementary Table S1. Improved cytotoxic potency following affinity optimization of 5T4 antibody; Supplementary Table S2. Quantification of cell surface-associated 5T4 in various cancer cell lines;Supplementary Table S3. Tumorigenicity of CSC populations derived from NCI- N87 and MDA-MB-361 xenografts; Supplementary Materials & Methods



Antibody–drug conjugates (ADC) are used to selectively deliver cytotoxic agents to tumors and have the potential for increased clinical benefit to cancer patients. 5T4 is an oncofetal antigen overexpressed on the cell surface in many carcinomas on both bulk tumor cells as well as cancer stem cells (CSC), has very limited normal tissue expression, and can internalize when bound by an antibody. An anti-5T4 antibody was identified and optimized for efficient binding and internalization in a target-specific manner, and engineered cysteines were incorporated into the molecule for site-specific conjugation. ADCs targeting 5T4 were constructed by site-specifically conjugating the antibody with payloads that possess different mechanisms of action, either a DNA cross-linking pyrrolobenzodiazepine (PBD) dimer or a microtubule-destabilizing tubulysin, so that each ADC had a drug:antibody ratio of 2. The resulting ADCs demonstrated significant target-dependent activity in vitro and in vivo; however, the ADC conjugated with a PBD payload (5T4-PBD) elicited more durable antitumor responses in vivo than the tubulysin conjugate in xenograft models. Likewise, the 5T4-PBD more potently inhibited the growth of 5T4-positive CSCs in vivo, which likely contributed to its superior antitumor activity. Given that the 5T4-PBD possessed both potent antitumor activity as well as anti-CSC activity, and thus could potentially target bulk tumor cells and CSCs in target-positive indications, it was further evaluated in non-GLP rat toxicology studies that demonstrated excellent in vivo stability with an acceptable safety profile. Taken together, these preclinical data support further development of 5T4-PBD, also known as MEDI0641, against 5T4+ cancer indications. Mol Cancer Ther; 16(8); 1576–87. ©2017 AACR.