154P - Computer-aided drug design based on CLDN4 ligand and its biological evaluation in ovarian cancer

Background

Numerous studies have consistently highlighted CLDN4 as a viable candidate for targeted therapy across various ovarian tumor types. Specifically, the C-terminal region of Clostridium perfringens enterotoxin (CPE) has demonstrated the ability to establish a stable complex with CLDN4. In our investigation, we harnessed computer-aided drug design (CADD) technology to craft peptides characterized by a specific and robust affinity for CLDN4. These peptides hold the potential to facilitate the development of self-assembling anti-tumor Peptide-Drug Conjugates (PDCs).

Methods

We employed virtual screening techniques, including alanine mutation, saturation mutation, and multi-point mutation, in conjunction with DS software to screen peptide sequences. To validate the binding capacity of the selected peptides with the CLDN4 protein, we conducted Surface Plasmon Resonance (SPR) and immunofluorescence colocalization experiments. Subsequently, we prepared Peptide-Drug Conjugates (PDCs), taking advantage of their inherent hydrophilic and hydrophobic properties, which promote their spontaneous assembly into nanofibrous structures. The anti-tumor efficacy of these formulations was rigorously assessed through both in vivo and in vitro experiments.

Results

SPR analysis revealed a noteworthy binding affinity between the targeted peptide and the CLDN4 protein, as evidenced by a Kd value of 5.343nM. Immunofluorescence co-localization experiments unequivocally demonstrated the co-localization of the targeted peptide with CLDN4. Furthermore, The PDC self-assembly group exhibited superior cytotoxicity compared to both the PDC non-self-assembly group and the camptothecin group.

Conclusions

In this study, we harnessed Computer-Aided Drug Design (CADD) technology to effectively engineer a targeting peptide characterized by a strong affinity for the CLDN4 protein. This peptide was subsequently employed in the self-assembly of Protein-Drug Conjugates (PDCs). The therapeutic potential of these PDCs for ovarian cancer was robustly substantiated through a comprehensive array of in vitro and in vivo experiments, unequivocally affirming their promise as a viable treatment option.

Legal entity responsible for the study

The authors.

Funding

National Natural Science Foundation of China.

Disclosure

All authors have declared no conflicts of interest.