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Tumor-selective blockade of CD47 signaling with a CD47/PD-L1 bispecific antibody for enhanced anti-tumor activity and limited toxicity

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Abstract

CD47, an immune checkpoint receptor frequently unregulated in various blood and solid tumors, interacts with ligand SIPRα on innate immune cells, and conveys a “do not eat me” signal to inhibit macrophage-mediated tumor phagocytosis. This makes CD47 a valuable target for cancer immunotherapy. However, the therapeutic utility of CD47-SIRPα blockade monoclonal antibodies is largely compromised due to significant red blood cell (RBCs) toxicities and fast target-mediated clearance as a result of extensive expression of CD47 on normal cells. To overcome these limitations and further improve therapeutic efficacy, we designed IBI322, a CD47/PD-L1 bispecific antibody which attenuated CD47 activity in monovalent binding and blocked PD-L1 activity in bivalent binding. IBI322 selectively bound to CD47+PD-L1+ tumor cells, effectively inhibited CD47-SIRPα signal and triggered strong tumor cell phagocytosis in vitro, but only with minimal impact on CD47 single positive cells such as human RBCs. In addition, as a dual blocker of innate and adaptive immune checkpoints, IBI322 effectively accumulated in PD-L1-positive tumors and demonstrated synergistic activity in inducing complete tumor regression in vivo. Furthermore, IBI322 showed only marginal RBCs depletion and was well tolerated in non-human primates (NHP) after repeated weekly injections, suggesting a sufficient therapeutic window in future clinical development of IBI322 for cancer treatment.

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Abbreviations

ATCC:

American Type Culture Collection

DCs:

Dendritic cells

DFO:

P-SCN-Deferoxamine

IBI322:

An anti-CD47/PD-L1 bispecific antibody

NHPs:

Non-human primates

PD-L1:

Programmed death-ligand 1

PD-1:

Programmed death-1

RBCs:

Red blood cells

SIRPα:

Signal-regulatory protein-α

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Funding

This work was financially supported by the National Significant New Drugs Creation Program (2017ZX09304021), Jiangsu Provincial Medical Innovation Team (CXTDA2017024), and Leading technology foundation research project of Jiangsu province (BK20192005).

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Authors and Affiliations

  1. Department of Clinical Pharmacology, First Affiliated Hospital of Soochow University, 899 Pinghai Road, Gusu District, Suzhou, 215006, Jiangsu Province, China

    Yan Wang, Chenrong Huang, Yicong Bian & Liyan Miao

  2. NHC Key Laboratory of Nuclear Medicine, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, 20 Qianrong Road, Binhu District, Wuxi, 214063, Jiangsu Province, China

    Yan Wang, Donghui Pan, Mingzhu Li & Min Yang

  3. Institute for Interdisciplinary Drug Research and Translational Sciences, College of Pharmaceutical Sciences, Soochow University, Suzhou, China

    Yan Wang, Chenrong Huang, Yicong Bian & Liyan Miao

  4. Innovent Biologics (Suzhou) Co., Ltd., 168 Dongping Street, Suzhou Industrial Park, Suzhou, 215123, Jiangsu Province, China

    Haiqing Ni, Shuaixiang Zhou, Kaijie He, Yarong Gao, Weiwei Wu, Min Wu, Zhihai Wu, Xuan Qiu, Ying Zhou, Bingliang Chen & Junjian Liu

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  1. Yan Wang
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Contributions

YW, HN, SZ, BC, MY, LM and JL designed the study; HN, YG, SZ, WW and ZW performed the in vitro experiment; WW, MW, YZ, BC, YW, DP, CH, ML, and YB performed the in vivo experiment and biodistribution; XQ performed the experiment in primate. HN, SZ, KH and JL wrote the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Min Yang, Liyan Miao or Junjian Liu.

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Conflict of interest

Haiqing Ni, Shuaixiang Zhou, Kaijie He, Yarong Gao, Weiwei Wu, Min Wu, Zhihai Wu, Xuan Qiu, ying Zhou, Bingliang Chen, and Junjian Liu are employees of Innovent Biologics (Suzhou). All the remaining authors have declared no conflicts of interest.

Ethical approval

All mice experiments were performed in accordance with the regulations for care and use of laboratory animals at Innovent Biologics and were approved by Innovent’s Institutional Animal Care and Use Committee (IACUC-01). All monkey experiments were approved by IACUC and performed according to the regulation of AAALAC.

Animal source

NOS-SCID mice and NOG mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (Beijing, China). Human CD47 knock-in mice were purchased from Biocytogen, Inc. (Beijing, China). Cynomolgus monkeys were purchased from Beijing Prima Biotech Inc.

Cell line authentication

RAJI, A375, CCRF-CEM, and H292 cell lines were obtained from ATCC (Manassas, VA). CHO-S cell line was obtained from Themo Fisher Scientific, Carlsbad, CA, USA). MC38 cell line was obtained from Shanghai Model Organisms Center, Inc. (Shanghai, China). PBMC cells were purchased from Allcells (Alameda, CA, USA).

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Yan Wang, Haiqing Ni and Shuaixiang Zhou contributed equally to this work.

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Wang, Y., Ni, H., Zhou, S. et al. Tumor-selective blockade of CD47 signaling with a CD47/PD-L1 bispecific antibody for enhanced anti-tumor activity and limited toxicity. Cancer Immunol Immunother 70, 365–376 (2021). https://doi.org/10.1007/s00262-020-02679-5

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