Chemoproteomic identification of molecular targets of antifungal prototypes, thiosemicarbazide and a camphene derivative of thiosemicarbazide, in Paracoccidioides brasiliensis

• Published in: PloS one 2018-08, Vol.13 (8), p.e0201948
• Main Authors: Borba, Joyce Villa Verde Bastos, Tauhata, Sinji Borges Ferreira, Oliveira, Cecília Maria Alves de, Ferreira Marques, Monique, Bailão, Alexandre Melo, Soares, Célia Maria de Almeida, Pereira, Maristela
• Format: Article
• Language: English
• Subjects: Amidohydrolases - metabolism; Amphotericin B; Animals; Antifungal agents; Antifungal Agents - chemistry; Antifungal Agents - isolation & purification; Antifungal Agents - pharmacology; Apoptosis; BALB 3T3 Cells; Biology and Life Sciences; Camphene; Carbohydrates; Cell cycle; Cell division; Cell Survival - drug effects; Chemical properties; Dimorphism; Drug Discovery; Drug therapy; Enzyme Activation - drug effects; Fluconazole; Formamidase; Fungal infections; Fungal Proteins - antagonists & inhibitors; Fungal Proteins - metabolism; Fungi; Fungicides; Glycolysis; Humans; Identification and classification; Itraconazole; Ketoconazole; Lungs; Mice; Microbial Sensitivity Tests; Mode of action; Morphogenesis; Organs; Paracoccidioides - drug effects; Paracoccidioides - metabolism; Paracoccidioidomycosis; Peptidases; Phylogenetics; Proteasomes; Protein Binding; Proteins; Proteomics; Proteomics - methods; Prototypes; Semicarbazides - chemistry; Semicarbazides - isolation & purification; Semicarbazides - pharmacology; South American blastomycosis; Sulfadiazine; Sulfamethoxazole; Target recognition; Thiosemicarbazides; Trimethoprim; Trimethoprim-sulfamethoxazole; Yeast; Yeasts
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0201948

Paracoccidioidomycosis (PCM) is a neglected human systemic disease caused by species of the genus Paracoccidioides. The disease attacks the host's lungs and may disseminate to many other organs. Treatment involves amphotericin B, sulfadiazine, trimethoprim-sulfamethoxazole, itraconazole, ketoconazole, or fluconazole. The treatment duration is usually long, from 6 months to 2 years, and many adverse effects may occur in relation to the treatment; co-morbidities and poor treatment adherence have been noted. Therefore, the discovery of more effective and less toxic drugs is needed. Thiosemicarbazide (TSC) and a camphene derivative of thiosemicarbazide (TSC-C) were able to inhibit P. brasiliensis growth at a low dosage and were not toxic to fibroblast cells. In order to investigate the mode of action of those compounds, we used a chemoproteomic approach to determine which fungal proteins were bound to each of these compounds. The compounds were able to inhibit the activities of the enzyme formamidase and interfered in P. brasiliensis dimorphism. In comparison with the transcriptomic and proteomic data previously obtained by our group, we determined that TSC and TSC-C were multitarget compounds that exerted effects on the electron-transport chain and cell cycle regulation, increased ROS formation, inhibited proteasomes and peptidases, modulated glycolysis, lipid, protein and carbohydrate metabolisms, and caused suppressed the mycelium to yeast transition.