Preclinical characterization and comparison between CD3/CD19 bispecific and novel CD3/CD19/CD20 trispecific antibodies against B-cell acute lymphoblastic leukemia: targeted immunotherapy for acute lymphoblastic leukemia

Abstract

The CD19-targeting bispecific T-cell engager blinatumomab has shown remarkable efficacy in patients with relapsed/refractory B-cell precursor acute lymphoblastic leukemia. However, several studies showed that blinatumomab has a short plasma half-life due to its low molecular weight, and thus its clinical use is limited. Furthermore, multiple trials have shown that approximately 30% of blinatumomab-relapsed cases are characterized by CD19 negative leukemic cells. Here, we design and characterize two novel antibodies, A-319 and A-2019. Blinatumomab and A-319 are CD3/CD19 bispecific antibodies with different molecular sizes and structures, and A-2019 is a novel CD3/CD19/CD20 trispecific antibody with an additional anti-CD20 function. Our in vitro, ex vivo, and in vivo experiments demonstrated that A-319 and A-2019 are potent antitumor agents and capable of recruiting CD3 positive T cells, enhancing T-cell function, mediating B-cell depletion, and eventually inhibiting tumor growth in Raji xenograft models. The two molecules are complementary in terms of efficacy and specificity profile. The activity of A-319 demonstrated superior to that of A-2019, whereas A-2019 has an additional capability to target CD20 in cells missing CD19, suggesting its potential function against CD19 weak or negative CD20 positive leukemic cells.

Keywords

B-cell acute lymphoblastic leukemia / bispecific antibody / trispecific antibody / CD19 / CD20

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Acknowledgements

This study was funded by the National Natural Science Foundation of China (Nos. 81670147, 81570178, and Antrag M-0377), Shanghai Municipal Education Commission-Major Project for Scientific Research and Innovation Plan of Natural Science (No. 2021-01-07-00-02-E00091), and Gaofeng Clinical Medicine Grant Support of Shanghai Municipal Education (No. 20172002).

Compliance with ethics guidelines

Xiaoqiang Yan, Stewart Leung, and Yushan Kong are employees of Generon Biomed Shanghai dedicated to the development of bispecific antibodies for immunotherapy. Sisi Wang, Lijun Peng, Wenqian Xu, Yuebo Zhou, Ziyan Zhu, Jin Wang, and Jian-Qing Mi declare no competing financial interests. All procedures performed in studies involving human blood samples were approved by the ethics committee at Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine and Shanghai Blood Center and performed according to the principles of the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards. All institutional and national guidelines for the care and use of laboratory animals were followed. All patients and healthy blood donors gave written informed consent prior to blood sample collection for the use of biomaterials and clinical data for scientific purposes.

Electronic Supplementary Material

Supplementary material is available in the online version of this article at https://doi.org/10.1007/s11684-021-0835-8 and is accessible for authorized users.

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