Discovery and optimization of a novel anti-GUCY2c x CD3 bispecific antibody for the treatment of solid tumors. Issue 1 (1st January 2021)

• Record Type: Journal Article
• Authors: Root, Adam R.; Guntas, Gurkan; Katragadda, Madan; Apgar, James R.; Narula, Jatin; Chang, Chew Shun; Hanscom, Sara; McKenna, Matthew; Wade, Jason; Meade, Caryl; Ma, Weijun; Guo, Yongjing; Liu, Yan; Duan, Weili; Hendershot, Claire; King, Amy C.; Zhang, Yan; Sousa, Eric; Tam, Amy; Benard, Susan; Yang, Han; Kelleher, Kerry; Jin, Fang; Piche-Nicholas, Nicole; Keating, Sinead E.; Narciandi, Fernando; Lawrence-Henderson, Rosemary; Arai, Maya; Stochaj, Wayne R.; Svenson, Kristine; Mosyak, Lidia; Lam, Khetemcnee; Francis, Christopher; Marquette, Kimberly; Wroblewska, Liliana; Zhu, H. Lily; Sheehan, Alfredo Darmanin; LaVallie, Edward R.; D'Antona, Aaron M.; Betts, Alison; King, Lindsay; Rosfjord, Edward; Cunningham, Orla; Lin, Laura; Sapra, Puja; Tchistiakova, Lioudmila; Mathur, Divya; Bloom, Laird;
• Is Part Of: MAbs. Volume 13:Issue 1(2021)
• Publication Date: 2021-01-01
• Subjects: T cell bispecific -- T cell retargeting -- T-BsAb -- immuno-oncology -- GUCY2C -- Guanlyate cyclase 2c (GCC) -- PF-07062119 -- antibody engineering -- antibody optimization -- developability -- high-throughput protein production; Monoclonal antibodies -- Therapeutic use -- Periodicals; Monoclonal antibodies -- Periodicals; Antibodies, Monoclonal -- Periodicals; 616.0798
• DOI: 10.1080/19420862.2020.1850395
• Languages: English
• ISSNs: 1942-0862

Abstract

ABSTRACT: We report here the discovery and optimization of a novel T cell retargeting anti-GUCY2C x anti-CD3ε bispecific antibody for the treatment of solid tumors. Using a combination of hybridoma, phage display and rational design protein engineering, we have developed a fully humanized and manufacturable CD3 bispecific antibody that demonstrates favorable pharmacokinetic properties and potent in vivo efficacy. Anti-GUCY2C and anti-CD3ε antibodies derived from mouse hybridomas were first humanized into well-behaved human variable region frameworks with full retention of binding and T-cell mediated cytotoxic activity. To address potential manufacturability concerns, multiple approaches were taken in parallel to optimize and de-risk the two antibody variable regions. These approaches included structure-guided rational mutagenesis and phage display-based optimization, focusing on improving stability, reducing polyreactivity and self-association potential, removing chemical liabilities and proteolytic cleavage sites, and de-risking immunogenicity. Employing rapid library construction methods as well as automated phage display and high-throughput protein production workflows enabled efficient generation of an optimized bispecific antibody with desirable manufacturability properties, high stability, and low nonspecific binding. Proteolytic cleavage and deamidation in complementarity-determining regions were also successfully addressed. Collectively, these improvements translated to a molecule with potent single-agent in vivo efficacy in a tumor cell line adoptive transfer model and a cynomolgus monkey pharmacokinetic profile (half-life>4.5 days) suitable for clinical development. Clinical evaluation of PF-07062119 is ongoing.