Mo-CBP4, a purified chitin-binding protein from Moringa oleifera seeds, is a potent antidermatophytic protein: In vitro mechanisms of action, in vivo effect against infection, and clinical application as a hydrogel for skin infection

Abstract

Dermatophytes belonging to Trichophyton ssp. are important anthropophilic and zoophilic pathogens, which developed resistance to griseofulvin, the common antifungal drug used to treat dermatophytosis. In this context, Moringa oleifera seed proteins have been described as antifungal agents with potential applications. Thus, this work aimed to evaluate the antidermatophytic in vitro, focusing on mechanisms, and in vivo potential of Mo-CBP₄, purified from M. oleifera seeds. Mo-CBP₄was purified after protein extraction with 50 mM Tris-HCl buffer, pH 8.0, and chromatography on chitin and CM Sepharose™ columns and antidermatophytic potential of Mo-CBP₄ evaluated in vitro and in vivo. In vitro, Mo-CBP₄ reduced in 50% the germination of microconidia of Trichophyton mentagrophytes at 45 μM; but did not show inhibition of mycelial growth. Mo-CBP₄ (45 μM) presents the inhibitory activity even when incubated with N-acetyl-d-glucosamine (NAG). Analysis of the mechanisms of Mo-CBP₄ revealed an increase in membrane permeability, ROS overproduction and damage to cell wall leading to microconidia death. Furthermore, using in vivo models, Mo-CBP₄ (5, 10 and 20 mg g^-1) reduced the severity and time of dermatophytosis. Altogether, these findings indicate that Mo-CBP₄ has great potential for the development of novel antifungal drugs for the clinical treatment of dermatophytosis.

Keywords: Anti-dermatophytic potential; Chitin-binding protein; Moringa oleifera.