mAbs (Jan 2021)

Discovery and optimization of a novel anti-GUCY2c x CD3 bispecific antibody for the treatment of solid tumors

  • Adam R. Root,
  • Gurkan Guntas,
  • Madan Katragadda,
  • James R. Apgar,
  • Jatin Narula,
  • Chew Shun Chang,
  • Sara Hanscom,
  • Matthew McKenna,
  • Jason Wade,
  • Caryl Meade,
  • Weijun Ma,
  • Yongjing Guo,
  • Yan Liu,
  • Weili Duan,
  • Claire Hendershot,
  • Amy C. King,
  • Yan Zhang,
  • Eric Sousa,
  • Amy Tam,
  • Susan Benard,
  • Han Yang,
  • Kerry Kelleher,
  • Fang Jin,
  • Nicole Piche-Nicholas,
  • Sinead E. Keating,
  • Fernando Narciandi,
  • Rosemary Lawrence-Henderson,
  • Maya Arai,
  • Wayne R. Stochaj,
  • Kristine Svenson,
  • Lidia Mosyak,
  • Khetemcnee Lam,
  • Christopher Francis,
  • Kimberly Marquette,
  • Liliana Wroblewska,
  • H. Lily Zhu,
  • Alfredo Darmanin Sheehan,
  • Edward R. LaVallie,
  • Aaron M. D’Antona,
  • Alison Betts,
  • Lindsay King,
  • Edward Rosfjord,
  • Orla Cunningham,
  • Laura Lin,
  • Puja Sapra,
  • Lioudmila Tchistiakova,
  • Divya Mathur,
  • Laird Bloom

DOI
https://doi.org/10.1080/19420862.2020.1850395
Journal volume & issue
Vol. 13, no. 1

Abstract

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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.

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