Mechanism Study of Thermally Induced Anti-Tumor Drug Loading to Engineered Human Heavy-Chain Ferritin Nanocages Aided by Computational Analysis

• Published in: Biosensors (Basel) Vol. 11; no. 11; p. 444
• Main Authors: Yin, Shuang, Liu, Yongdong, Dai, Sheng, Zhang, Bingyang, Qu, Yiran, Zhang, Yao, Choe, Woo-Seok, Bi, Jingxiu
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 11-11-2021; MDPI
• Subjects: Antineoplastic Agents - administration & dosage; Chains; Chromatography; computational analysis; Computer applications; Doxorubicin; Doxorubicin - administration & dosage; drug delivery; Drugs; E coli; Ferritin; Ferritins - chemistry; Humans; In vitro methods and tests; Investigations; Ligands; Molecular docking; Molecular Docking Simulation; Molecular dynamics; Nanoparticles; Neoplasms - drug therapy; Optimization; Peptides; Proteins; Recovery; Simulation; Size exclusion chromatography; thermally induced drug loading; Transmission electron microscopy; Tumors; Australia; United States > US; ferritin; computational analysis; drug delivery; thermally induced drug loading
• ISSN: 2079-6374; 2079-6374
• DOI: 10.3390/bios11110444

Diverse drug loading approaches for human heavy-chain ferritin (HFn), a promising drug nanocarrier, have been established. However, anti-tumor drug loading ratio and protein carrier recovery yield are bottlenecks for future clinical application. Mechanisms behind drug loading have not been elaborated. In this work, a thermally induced drug loading approach was introduced to load anti-tumor drug doxorubicin hydrochloride (DOX) into HFn, and 2 functionalized HFns, HFn-PAS-RGDK, and HFn-PAS. Optimal conditions were obtained through orthogonal tests. All 3 HFn-based proteins achieved high protein recovery yield and drug loading ratio. Size exclusion chromatography (SEC) and transmission electron microscopy (TEM) results showed the majority of DOX loaded protein (protein/DOX) remained its nanocage conformation. Computational analysis, molecular docking followed by molecular dynamic (MD) simulation, revealed mechanisms of DOX loading and formation of by-product by investigating non-covalent interactions between DOX with HFn subunit and possible binding modes of DOX and HFn after drug loading. In in vitro tests, DOX in protein/DOX entered tumor cell nucleus and inhibited tumor cell growth.