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A novel human single-domain antibody-drug conjugate targeting CEACAM5 exhibits potent in vitro and in vivo antitumor activity

Acta Pharmacologica Sinica volume 45, pages 609–618 (2024)Cite this article

Abstract

Leveraging the specificity of antibody to deliver cytotoxic agent into tumor, antibody-drug conjugates (ADCs) have become one of the hotspots in the development of anticancer therapies. Although significant progress has been achieved, there remain challenges to overcome, including limited penetration into solid tumors and potential immunogenicity. Fully human single-domain antibodies (UdAbs), with their small size and human nature, represent a promising approach for addressing these challenges. Carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5) is a glycosylated cell surface protein that rarely expressed in normal adult tissues but overexpressed in diverse cancers, taking part in tumorigenesis, progression, and metastasis. In this study, we investigated the therapeutic potential of UdADC targeting CEACAM5. We performed biopanning in our library and obtained an antibody candidate B9, which bound potently and specifically to CEACAM5 protein (KD = 4.84 nM) and possessed excellent biophysical properties (low aggregation tendency, high homogeneity, and thermal stability). The conjugation of B9 with a potent cytotoxic agent, monomethyl auristatin E (MMAE), exhibited superior antitumor efficacy against CEACAM5-expressing human gastric cancer cell line MKN-45, human pancreatic carcinoma cell line BxPC-3 and human colorectal cancer cell line LS174T with IC50 values of 38.14, 25.60, and 101.4 nM, respectively. In BxPC-3 and MKN-45 xenograft mice, administration of UdADC B9-MMAE (5 mg/kg, i.v.) every 2 days for 4 times markedly inhibited the tumor growth without significant change in body weight. This study may have significant implications for the design of next-generation ADCs.

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Fig. 1: Isolation of a human single-domain antibody targeting CEACAM5.
Fig. 2: Biophysical characteristics of UdAb B9.
Fig. 3: Molecular and cellular binding profile of UdAb B9 to the CEACAM5 protein.
Fig. 4: In vitro cell cytotoxicity of the UdADC B9-MMAE.
Fig. 5: In vivo anti-tumor efficacy of UdADC B9-MMAE.
Fig. 6: Safety profile of the UdADC B9-MMAE in BALB/c mice.

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Acknowledgements

This project was supported by National Key R&D Program of China (2019YFA0904400), and National Natural Science Foundation of China (32270984).

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Author notes

  1. These authors contributed equally: Xiao-yi Zhu, Quan-xiao Li

Authors and Affiliations

  1. MOE/NHC/CAMS Key Laboratory of Medical Molecular Virology, Shanghai Frontiers Science Center of Pathogenic Microorganisms and Infection, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai, 200032, China

    Xiao-yi Zhu, Quan-xiao Li, Yu Kong, Ke-ke Huang, Gang Wang, Yun-ji Wang, Yan-ling Wu & Tian-lei Ying

  2. Shanghai Engineering Research Center for Synthetic Immunology, Shanghai, 200032, China

    Xiao-yi Zhu, Quan-xiao Li, Yu Kong, Ke-ke Huang, Yan-ling Wu & Tian-lei Ying

  3. Auckland Bioengineering Institute, University of Auckland, Auckland, 1010, New Zealand

    Jun Lu

  4. Cancer Institute, Fudan University Shanghai Cancer Center, Shanghai, 200032, China

    Guo-qiang Hua

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  1. Xiao-yi Zhu
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Contributions

XYZ and QXL performed the experiments, analyzed the data and conceived the manuscript with the help of YK, KKH, GW, and YJW. TLY and YLW initiated, planned, and supervised the project. JL and GQH revised and edited the final version of the manuscript. All authors reviewed and approved the submission version of the manuscript.

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Correspondence to Yan-ling Wu or Tian-lei Ying.

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Zhu, Xy., Li, Qx., Kong, Y. et al. A novel human single-domain antibody-drug conjugate targeting CEACAM5 exhibits potent in vitro and in vivo antitumor activity. Acta Pharmacol Sin 45, 609–618 (2024). https://doi.org/10.1038/s41401-023-01200-9

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