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
Affiliations
- Adam R. Root
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Gurkan Guntas
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Madan Katragadda
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- James R. Apgar
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Jatin Narula
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Chew Shun Chang
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Sara Hanscom
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Matthew McKenna
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Jason Wade
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Caryl Meade
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Weijun Ma
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Yongjing Guo
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Yan Liu
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Weili Duan
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Claire Hendershot
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Amy C. King
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Yan Zhang
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Eric Sousa
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Amy Tam
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Susan Benard
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Han Yang
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Kerry Kelleher
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Fang Jin
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Nicole Piche-Nicholas
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Sinead E. Keating
- BioMedicine Design, Pfizer Inc., Dublin, IE, USA
- Fernando Narciandi
- BioMedicine Design, Pfizer Inc., Dublin, IE, USA
- Rosemary Lawrence-Henderson
- BioMedicine Design, Pfizer Inc., Andover, MA, USA
- Maya Arai
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Wayne R. Stochaj
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Kristine Svenson
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Lidia Mosyak
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Khetemcnee Lam
- BioMedicine Design, Pfizer Inc., Andover, MA, USA
- Christopher Francis
- BioMedicine Design, Pfizer Inc., Andover, MA, USA
- Kimberly Marquette
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Liliana Wroblewska
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- H. Lily Zhu
- BioMedicine Design, Pfizer Inc., Andover, MA, USA
- Alfredo Darmanin Sheehan
- BioMedicine Design, Pfizer Inc., Dublin, IE, USA
- Edward R. LaVallie
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Aaron M. D’Antona
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Alison Betts
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Lindsay King
- BioMedicine Design, Pfizer Inc., Andover, MA, USA
- Edward Rosfjord
- Oncology Research & Development, Pfizer Inc., Pearl River, NY, USA
- Orla Cunningham
- BioMedicine Design, Pfizer Inc., Dublin, IE, USA
- Laura Lin
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Puja Sapra
- Oncology Research & Development, Pfizer Inc., Pearl River, NY, USA
- Lioudmila Tchistiakova
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- Divya Mathur
- Oncology Research & Development, Pfizer Inc., Pearl River, NY, USA
- Laird Bloom
- BioMedicine Design, Pfizer Inc., Cambridge, MA, USA
- DOI
- https://doi.org/10.1080/19420862.2020.1850395
- Journal volume & issue
-
Vol. 13,
no. 1
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.
Keywords