RESUMO
Antimicrobial peptides (AMP) represent an alternative in the treatment of fungal infections associated with countless deaths. Here, we report a new AMP, named KWI-19, which was designed based on a peptide encrypted in the sequence of an Inga laurina Kunitz-type inhibitor (ILTI). KWI-19 inhibited the growth of Candida species and acted as a fungicidal agent from 2.5 to 20 µmol L-1, also showing synergistic activity with amphotericin B. Kinetic assays showed that KWI-19 killed Candida tropicalis cells within 60 min. We also report the membrane-associated mechanisms of action of KWI-19 and its interaction with ergosterol. KWI-19 was also characterized as a potent antibiofilm peptide, with activity against C. tropicalis. Finally, non-toxicity was reported against Galleria mellonella larvae, thus strengthening the interest in all the bioactivities mentioned above. This study extends our knowledge on how AMPs can be engineered from peptides encrypted in larger proteins and their potential as candicidal agents.
Assuntos
Antifúngicos , Candida , Animais , Antifúngicos/farmacologia , Anfotericina B/farmacologia , Peptídeos/farmacologia , Candida tropicalis , Inibidores de Proteases , Peptídeo HidrolasesRESUMO
Antimicrobial peptides (AMP) are present in all organisms and can present several activities and potential applications in human and animal health. Screening these molecules scaffolds represents a key point for discovering and developing novel biotechnological products, including antimicrobial, antiviral and anticancer drugs candidates and insecticidal molecules with potential applications in agriculture. Therefore, considering the amount of biological data currently deposited on public databases, computational approaches have been commonly used to predicted and identify novel cysteine-rich peptides scaffolds with known or unknown biological properties. Here, we describe a step-by-step in silico screening for cysteine-rich peptides employing molecular modeling (with a core focus on comparative modeling) and atomistic molecular dynamics simulations. Moreover, we also present the concept of additional tools aiming at the computer-aided screening of new Cs-AMPs based drug candidates. After the computational screening and peptide chemical synthesis, we also provide the reader with a step-by-step in vitro activity evaluation of these candidates, including antibacterial, antifungal, and antiviral assays.