Encapsulation of copper phenanthroline within horse spleen apoferritin: characterisation, cytotoxic activity and ability to retain temozolomide

Cassioli, Maria Letizia; Fay, Michael; Turyanska, Lyudmila; Bradshaw, Tracey D.; Thomas, Neil R.; Pordea, Anca

Abstract

Protein capsules are promising drug delivery vehicles for cancer research therapies. Apoferritin (AFt) is a self-assembling 12 nm diameter hollow nanocage with many desirable features for drug delivery, however, control of drug retention inside the protein cage remains challenging. Here we report the encapsulation of copper(II)-1,10-phenanthroline (Cu(phen)) within the horse spleen AFt (HSAFt) nanocage, by diffusion of the metal through the pores between the protein subunits. Transmission electron microscopy revealed the formation of organised copper adducts inside HSAFt, without affecting protein integrity. These structures proved stable during storage (>4 months at −20 °C). Exposure to physiologically relevant conditions (37 °C) showed some selectivity in cargo release after 24 h at pH 5.5, relevant to the internalisation of AFt within the endosome (60% release), compared to pH 7.4, relevant to the bloodstream (40% release). Co-encapsulation of temozolomide, a prodrug used to treat glioblastoma multiforme, and Cu(phen) enabled entrapment of an average of 339 TMZ molecules per cage. In vitro results from MTT and clonogenic assays identified cytotoxic activity of the Cu(phen), HSAFt–Cu(phen) and HSAFt–Cu(phen)–TMZ adducts against colorectal cancer cells (HCT-116) and glioblastoma cells (U373V, U373M). However, the presence of the metal also contributed to more potent activity toward healthy MRC5 fibroblasts, a result that requires further investigation to assess the clinical viability of this system.

• Journal Article Type: Article
• Acceptance Date: Mar 13, 2024
• Online Publication Date: Apr 29, 2024
• Publication Date: 2024
• Deposit Date: Jun 6, 2024
• Publicly Available Date: Jun 7, 2024
• Journal: RSC Advances
• Electronic ISSN: 2046-2069
• Publisher: Royal Society of Chemistry
• Peer Reviewed: Peer Reviewed
• Volume: 14
• Issue: 20
• Pages: 14008-14016
• DOI: https://doi.org/10.1039/d3ra07430g
• Public URL: https://nottingham-repository.worktribe.com/output/34350521
• Publisher URL: https://pubs.rsc.org/en/content/articlelanding/2024/ra/d3ra07430g