Details

Autor(en) / Beteiligte

• de Oliveira, Pamella Christina Ortega
• Ceroullo, Millena Santana
• dos Santos, Mayane Barbosa
• Medeiros, Pedro Rodrigues Coelho
• Marques, Bruno Clemente Brandão
• Tinoco, Luzineide Wanderley
• de Souza, Maria Cecília Bastos Vieira
• da Costa Santos Boechat, Fernanda
• de Moraes, Marcela Cristina

Titel

Nucleoside Hydrolase immobilized on magnetic particles as a tool for onflow screening and characterization of inhibitors

Ist Teil von

• Journal of pharmaceutical and biomedical analysis, 2023-10, Vol.235, p.115589-115589, Article 115589

Ort / Verlag

England: Elsevier B.V

Erscheinungsjahr

2023

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• Nucleoside Hydrolases (NH) are considered a target for the development of new antiprotozoal agents. The development of new and automated screening assays for the identification of NH inhibitors can accelerate the first stages of the drug discovery process. In this work, NH from Leishmania donovani (LdNH) was covalently immobilized onto magnetic particles (LdNH-MPs) and trapped by magnets into a TFE tube to yield an immobilized enzyme reactor (IMER). For an automated assay, the LdNH-MP-IMER was connected in-line to an analytical column in an HPLC-DAD system to monitor the enzyme activity through quantification of the product hypoxanthine. Kinetic studies provided a KM value of 2,079 ± 87 µmol.L-1 for the inosine substrate. Validation of the LdNH-MP-IMER for onflow screening purposes was performed with a library containing 12 quinolone ribonucleosides. Among them, three were identified as new competitive LdNH inhibitors, with Ki values between 83.5 and 169.4 µmol.L-1. This novel in-line screening assay has proven to be reliable, fast, low cost, and applicable to large libraries of compounds. •The immobilization of LdNH on magnetic particles allowed the assembling of an IMER•Onflow monitoring of LdNH-IMER activity as an innovative approach•Development of an automated system to screen new inhibitors•Two quinolone ribonucleosides identified as new competitive inhibitors