Engineering a protease-stable, oral single-domain antibody to inhibit IL-23 signaling

N Naruhisa Ota (Department of Immunology Discovery, Genentech) C Christopher W. Davies (Antibody Engineering, Genentech) J Jing Kang (Translational Immunology, Genentech) D Donghong Yan (Translational Immunology, Genentech) A Alexis Scherl (Pathology, Genentech) A Anne Wong (Biochemical and Cellular Pharmacology, Genentech) R Ryan Cook (Biochemical and Cellular Pharmacology, Genentech) X Xun Tao (Preclinical and Translational Pharmacokinetics, Genentech) D Debra Dunlap (Pathology, Genentech) S Sha Klabunde (Translational Medicine, Genentech) P Priscilla Mantik (Small Molecule Pharmaceutics, Genentech) V Vishnu Mohanan (Translational Medicine, Genentech) W WeiYu Lin J Jacqueline McBride (Translational Medicine, Genentech) S Shraddha Sadekar (Preclinical and Translational Pharmacokinetics, Genentech) K Kelly M. Storek (Infectious Diseases, Genentech) P Patrick Lupardus (Structural Biology, Genentech) Z Zhengmao Ye (Biochemical and Cellular Pharmacology, Genentech) H Heidi Ackerly Wallweber (Structural Biology, Genentech) J James R. Kiefer (Structural Biology, Genentech) M Min Xu P Pamela Chan (Biochemical and Cellular Pharmacology, Genentech) K Karthik Nagapudi (Small Molecule Pharmaceutics, Genentech) T Tangsheng Yi J James T. Koerber (Antibody Engineering, Genentech)

Abstract

Interleukin (IL)-23 is a validated therapeutic target in inflammatory bowel disease. While antibodies targeting IL23 demonstrate clinical efficacy, they face challenges such as high costs, safety risks, and the necessity of parenteral administration. Here, we present a workflow to simultaneously enhance the affinity and protease stability of an inhibitory anti-IL23R VHH for oral use. Cocrystal structure analysis reveals that the anti-IL23R VHH employs both CDR and framework residues to achieve picomolar affinity for IL23R. The engineered VHH remains stable for over 8 h in intestinal fluid and 24 h in fecal samples. Oral administration of this VHH achieves deep pathway inhibition in a murine colitis model. Furthermore, a single pill provides sustained IL23R inhibition in nonhuman primate blood for over 24 h. With high potency, gut stability, high production yield, and favorable drug-like properties, oral VHHs offer a promising approach for inflammatory bowel diseases.

Article Details

Volume / Issue Vol. 122, Issue 22
Published June 03, 2025
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (25)

N

Naruhisa Ota

Department of Immunology Discovery, Genentech

C

Christopher W. Davies

Antibody Engineering, Genentech

J

Jing Kang

Translational Immunology, Genentech

D

Donghong Yan

Translational Immunology, Genentech

A

Alexis Scherl

Pathology, Genentech

A

Anne Wong

Biochemical and Cellular Pharmacology, Genentech

R

Ryan Cook

Biochemical and Cellular Pharmacology, Genentech

X

Xun Tao

Preclinical and Translational Pharmacokinetics, Genentech

D

Debra Dunlap

Pathology, Genentech

S

Sha Klabunde

Translational Medicine, Genentech

P

Priscilla Mantik

Small Molecule Pharmaceutics, Genentech

V

Vishnu Mohanan

Translational Medicine, Genentech

W

WeiYu Lin

J

Jacqueline McBride

Translational Medicine, Genentech

S

Shraddha Sadekar

Preclinical and Translational Pharmacokinetics, Genentech

K

Kelly M. Storek

Infectious Diseases, Genentech

P

Patrick Lupardus

Structural Biology, Genentech

Z

Zhengmao Ye

Biochemical and Cellular Pharmacology, Genentech

H

Heidi Ackerly Wallweber

Structural Biology, Genentech

J

James R. Kiefer

Structural Biology, Genentech

M

Min Xu

P

Pamela Chan

Biochemical and Cellular Pharmacology, Genentech

K

Karthik Nagapudi

Small Molecule Pharmaceutics, Genentech

T

Tangsheng Yi

J

James T. Koerber

Antibody Engineering, Genentech