Inhibition of Phospholipase by Orlistat as an Alternate Therapy to Combat Opportunistic Mycosis Caused by C. albicans.

Abstract

Candida albicans is one of the most important etiological agents causing an opportunistic mycosis, candidiasis. In the past, it was perceived to be associated with immunocompromised patients only. However, it has now been reported with several clinical complications with varying severity. Additionally, increasing incidences of multiple drug resistance associated with the infections have complicated its treatment as well. Therefore, an investigation of alternate therapy, for instance, inhibition of the virulence factors is desperately needed. In the present study, a multidrug-resistant Candida albicans SDL-4 was screened for secretion of the virulence factors: aspartyl proteases and phospholipases. The pathogen secreted phospholipases potentially compared to aspartyl proteases. Therefore, C. albicans SDL-4 phospholipase was purified to homogeneity, characterized, and its inhibition was studied subsequently. It catalysed the substrate, p-nitrophenyl palmitate, optimally in 0.1M acetate buffer, pH 5, at 37°C. In the present study, we also aimed to re-purpose orlistat, which is a commercially available anti-obesity drug. Orlistat, at the concentration of 360μg/ml, could diminish the activity and stability of the candidal virulence factor. Its half-life was reduced in the presence of orlistat at 37°C. As well, increase in Km and unaltered Vmax indicated that orlistat inhibited phospholipase competitively. The inhibition kinetics was supported by measuring alterations in the secondary structure of the candidal phospholipase upon treatment with orlistat by the circular dichroism spectroscopy and K2D3. Moreover, validation of the study at clinical level may establish orlistat as a supportive treatment to reduce invasiveness and related medical intricacies during candidiasis.