Antibody-mimetic drug conjugate with efficient internalization activity using anti-HER2 VHH and duocarmycin

doi:10.1016/j.pep.2023.106375

. 2024 Feb:214:106375.

doi: 10.1016/j.pep.2023.106375. Epub 2023 Oct 4.

Antibody-mimetic drug conjugate with efficient internalization activity using anti-HER2 VHH and duocarmycin

Juri Sakata¹, Toshifumi Tatsumi², Akira Sugiyama³, Akihiro Shimizu¹, Yuya Inagaki¹, Hiroto Katoh⁴, Takefumi Yamashita⁵, Kazuki Takahashi², Sho Aki⁶, Yudai Kaneko⁷, Takeshi Kawamura⁸, Mai Miura⁶, Masazumi Ishii⁶, Tsuyoshi Osawa⁶, Toshiya Tanaka⁶, Shumpei Ishikawa⁴, Masanobu Tsukagoshi⁹, Michael Chansler⁹, Tatsuhiko Kodama⁶, Motomu Kanai², Hidetoshi Tokuyama¹⁰, Kenzo Yamatsugu¹¹

Affiliations

¹ Graduate School of Pharmaceutical Sciences, Tohoku University, 6-3 Aoba, Aramaki, Aoba-ku, Sendai, 980-8578, Japan.
² Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
³ Isotope Science Center, The University of Tokyo, 2-11-16 Yayoi, Bunyo-ku, Tokyo, 113-0032, Japan; Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan. Electronic address: sugiyama@lsbm.org.
⁴ Department of Preventive Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
⁵ Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan; Department of Physical Chemistry, School of Pharmacy and Pharmaceutical Sciences, Hoshi University, 2-4-41 Ebara, Shinagawa-ku, Tokyo, 142-8501, Japan.
⁶ Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan.
⁷ Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan; Medical & Biological Laboratories Co., Ltd, 2-11-8 Shibadaimon, Minato-ku, Tokyo, 105-0012, Japan.
⁸ Isotope Science Center, The University of Tokyo, 2-11-16 Yayoi, Bunyo-ku, Tokyo, 113-0032, Japan; Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan.
⁹ Savid Therapeutics Inc., Eifuku 3-9-10, Suginami-ku, Tokyo, 168-0064, Japan.
¹⁰ Graduate School of Pharmaceutical Sciences, Tohoku University, 6-3 Aoba, Aramaki, Aoba-ku, Sendai, 980-8578, Japan. Electronic address: hidetoshi.tokuyama.d4@tohoku.ac.jp.
¹¹ Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba-shi, Chiba, 260-8675, Japan. Electronic address: yamatsugu@chiba-u.jp.

PMID: 37797818
DOI: 10.1016/j.pep.2023.106375

Free article

Antibody-mimetic drug conjugate with efficient internalization activity using anti-HER2 VHH and duocarmycin

Juri Sakata et al. Protein Expr Purif. 2024 Feb.

Free article

. 2024 Feb:214:106375.

doi: 10.1016/j.pep.2023.106375. Epub 2023 Oct 4.

Authors

Affiliations

¹ Graduate School of Pharmaceutical Sciences, Tohoku University, 6-3 Aoba, Aramaki, Aoba-ku, Sendai, 980-8578, Japan.
² Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
³ Isotope Science Center, The University of Tokyo, 2-11-16 Yayoi, Bunyo-ku, Tokyo, 113-0032, Japan; Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan. Electronic address: sugiyama@lsbm.org.
⁴ Department of Preventive Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
⁵ Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan; Department of Physical Chemistry, School of Pharmacy and Pharmaceutical Sciences, Hoshi University, 2-4-41 Ebara, Shinagawa-ku, Tokyo, 142-8501, Japan.
⁶ Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan.
⁷ Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan; Medical & Biological Laboratories Co., Ltd, 2-11-8 Shibadaimon, Minato-ku, Tokyo, 105-0012, Japan.
⁸ Isotope Science Center, The University of Tokyo, 2-11-16 Yayoi, Bunyo-ku, Tokyo, 113-0032, Japan; Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan.
⁹ Savid Therapeutics Inc., Eifuku 3-9-10, Suginami-ku, Tokyo, 168-0064, Japan.
¹⁰ Graduate School of Pharmaceutical Sciences, Tohoku University, 6-3 Aoba, Aramaki, Aoba-ku, Sendai, 980-8578, Japan. Electronic address: hidetoshi.tokuyama.d4@tohoku.ac.jp.
¹¹ Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba-shi, Chiba, 260-8675, Japan. Electronic address: yamatsugu@chiba-u.jp.

PMID: 37797818
DOI: 10.1016/j.pep.2023.106375

Abstract

Antibody-mimetic drug conjugate (AMDC) is a cancer cell-targeted drug delivery system based on the non-covalent binding of mutated streptavidin and modified biotin, namely Cupid and Psyche. However, the development of AMDCs is hampered by difficulties in post-translational modification or poor internalization activity. Here, we report an expression, refolding, and purification method for AMDC using a variable heavy chain of heavy chain-only antibodies (VHHs). Monomeric anti-HER2 VHH fused to Cupid was expressed in Escherichia coli inclusion bodies. Solubilization and refolding at optimized reducing conditions and pH levels were selected to form a functional, tetrameric protein (anti-HER2 VHH-Cupid) that can be easily purified based on molecular weight. Anti-HER2 VHH-Cupid non-covalently creates a tight complex with Psyche linked to a potent DNA-alkylating agent, duocarmycin. This complex can be absorbed by the HER2-expressing human breast cancer cell line, KPL-4, and kills KPL-4 cells in vitro and in vivo. The production of a targeting protein with internalizing activity, combined with the non-covalent conjugation of a highly potent payload, renders AMDC a promising platform for developing cancer-targeted therapy.

Keywords: Anti-tumor effects; Antibody-mimetic drug conjugate; Cupid-Psyche platform; Duocarmycin; KPL-4 cells; Non-covalent conjugation.

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Antibody-mimetic drug conjugate with efficient internalization activity using anti-HER2 VHH and duocarmycin

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Antibody-mimetic drug conjugate with efficient internalization activity using anti-HER2 VHH and duocarmycin

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