A Target Engagement Assay to Assess Uptake, Potency, and Retention of Antibiotics in Living Bacteria

Autor(es)

Fanti, Rebeka C. ; Vasconcelos, Stanley N. S. ; Catta-Preta, Carolina M. C. ; Sullivan, Jaryd R. . ; Riboldi, Gustavo P. ; Reis, Caio V. dos ; Ramos, Priscila Z. ; Edwards, Aled M. ; Behr, Marcel A. . ; Counago, Rafael M.

• Tipo de documento: Artigo Científico
• Fonte: ACS INFECTIOUS DISEASES; v. 8, n. 8, p. 19-pg., 2022-07-11.

Resumo

New antibiotics are urgently needed to counter the emergence of antimicrobial-resistant pathogenic bacteria. A major challenge in antibiotic drug discovery is to turn potent biochemical inhibitors of essential bacterial components into effective antimicrobials. This difficulty is underpinned by a lack of methods to investigate the physicochemical properties needed for candidate antibiotics to permeate the bacterial cell envelope and avoid clearance by the action of bacterial efflux pumps. To address these issues, here we used a target engagement assay to measure the equilibrium and kinetic binding parameters of antibiotics targeting dihydrofolate reductase (DHFR) in live bacteria. We also used this assay to identify novel DHFR ligands having antimicrobial activity . We validated this approach using the Gram-negative bacteria Escherichia coli and the emerging human pathogen Mycobacterium abscessus. We expect the use of target engagement assays in bacteria to expedite the discovery and progression of novel, cell-permeable antibiotics with on-target activity. (AU)

• Processo FAPESP: 13/50724-5 - Centro de Biologia Química de Proteínas Quinases: alavancando desenvolvimento de fármacos através de pesquisa de acesso aberto
• Beneficiário: Paulo Arruda
• Modalidade de apoio: Auxílio à Pesquisa - Parceria para Inovação Tecnológica - PITE
• Processo FAPESP: 14/50897-0 - INCT 2014: Centro de Química Medicinal de Acesso Aberto
• Beneficiário: Katlin Brauer Massirer
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 18/09475-5 - Novas abordagens para inibidores de proteínas quinases MRCKa, MRCKb E MRCKg
• Beneficiário: Stanley Nunes Siqueira Vasconcelos
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado