Design of Protease-Responsive Antifungal Liposomal Formulation Decorated with a Lipid-Modified Chitin-Binding Domain

• Published in: International journal of molecular sciences Vol. 25; no. 7; p. 3567
• Main Authors: Saputra, Hendra, Safaat, Muhammad, Santoso, Pugoh, Wakabayashi, Rie, Goto, Masahiro, Taira, Toki, Kamiya, Noriho
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.04.2024
• Subjects: AmBisome; Amphotericin B; Antifungal agents; Antifungal Agents - pharmacology; antifungal formulation; Antiparasitic agents; Aqueous solutions; C. albicans; Candida albicans; Cell growth; Chitin; Chitinase; Drug delivery systems; Drug resistance in microorganisms; Fungal infections; Fungi; Health aspects; lipid conjugation; Lipids; Liposomes; Peptide Hydrolases; Peptides; Peptides - pharmacology; Proteins; Thrombin; Toxicity; Japan; chitinase; antifungal formulation; lipid conjugation; C. albicans; AmBisome
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms25073567

is a prevalent fungal pathogen that displays antibiotic resistance. The polyene antifungal amphotericin B (AmB) has been the gold standard because of its broad antifungal spectra, and its liposomal formulation, AmBisome, has been used widely and clinically in treating fungal infections. Herein, we explored enhancing the antifungal activity of AmBisome by integrating a small chitin-binding domain (LysM) of chitinase A derived from . LysM conjugated with a lipid (LysM-lipid) was initially prepared through microbial transglutaminase (MTG)-mediated peptide tag-specific conjugation of LysM with a lipid-peptide substrate. The AmBisome formulation modified with LysM-lipid conjugates had a size distribution that was comparable to the native liposomes but an increased zeta potential, indicating that LysM-lipid conjugates were anchored to AmBisome. LysM-lipid-modified AmBisome exhibited long-term stability at 4 °C while retaining the capacity to bind chitin. Nevertheless, the antifungal efficacy of LysM-lipid-modified AmBisome against was modest. We then redesigned a new LysM-lipid conjugate by introducing a peptide linker containing a thrombin digestion (TD) site at the C-terminus of LysM (LysM-TD linker-lipid), thereby facilitating the liberation of the LysM domain from AmBisome upon the addition of thrombin. This new AmBisome formulation anchored with LysM-TD linker-lipid exhibited superior performance in suppressing growth in the presence of thrombin compared with the LysM-lipid formulation. These results provide a platform to design stimuli-responsive AmBisome formulations that respond to external environments and thus advance the treatment of pathogenic fungi infections.