De novo design of a two-step approach targeting Claudin-6 for enhanced drug delivery to solid tumors
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12967_2025_Article_7316.pdf
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Author(s)
Yan, Jiayao
Zhong, Liqing
Chen, Xiaotong
Li, Lin
Liu, Fangcen
Lei, Lei
An, Mengchao
Wei, Xiao
Wang, Ying
Chen, Tianran
Date Issued
November 20, 2025
Publisher
BioMed Central
Citation
Yan, J., Zhong, L., Chen, X. et al. De novo design of a two-step approach targeting Claudin-6 for enhanced drug delivery to solid tumors. J Transl Med 23, 1323 (2025).
Version
Final published version
Abstract
Background Although antibody-conjugated drugs have achieved success in clinical practice for cancer treatment, challenges remain in developing a highly efficient drug delivery system with specific accumulation in tumors and reduction in side effects. With improved pharmacokinetics, strong covalent bonding and quick binding reactions, a pre-targeting approach via molecular pairs represents an attractive platform for two-step delivery system construction. Methods Bioinformatics and immunohistochemistry assays were performed to assess Claudin-6 (CLDN6) as a highly specific tumor target in solid tumors. A phage-displayed library was used to screen and optimize anti-CLDN6 designed ankyrin repeat proteins (DARPins), which were incorporated into a two-step delivery system based on SpyTag/SpyCatcher. Fluorescent staining, flow cytometry and near-infrared imaging were performed to assess the tumor-targeting ability and biodistribution of this delivery system. The cytotoxic drug, Monomethyl auristatin E (MMAE), was conjugated with the delivery system to evaluate its anti-tumor efficacy and safety profile. Results Anti-CLDN6 DARPins exhibited specific binding to CLDN6+ cancer cells with high affinity instead of negative cells in vitro, ex vivo and in vivo. The DARPins-based two-step delivery system improved background clearance with a high signal-to-noise ratio, enhancing the specific accumulation of payloads in tumors. The cytotoxic drug delivered via the two-step system appeared superior to the one-step approach in IC50, biodistribution, and tumor growth inhibition. Conclusions Our study presented the de novo design of a two-step drug delivery system targeting Claudin-6 with enhanced anti-tumor efficacy and improved biosafety. These findings highlighted the potential of this approach to enhance the efficacy of tumor-targeting therapies and reduce adverse effects, paving the way for more effective cancer treatments.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Koch Institute for Integrative Cancer Research at MIT
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Creative Commons Attribution-NonCommercial-NoDerivatives
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DOI of Published Version
https://doi.org/10.1186/s12967-025-07316-2
