title: Immobilized cholinesterases capillary reactors on-flow screening of selective inhibitors.
abstract: The discovery of selective inhibitors for acetylcholinesterase (AChE) or butyrylcholinesterase (BChE) is extremely important for the development of drugs that can be used in the treatment of patients diagnosed with the Alzheimer's disease (AD). For this reason, there is a growing interest in developing rapid and effective assays techniques for cholinesterases (ChE) enzymes ligand screening. Herein is presented the results of selective screening assays of a coumarin derivatives library using BChE and AChE covalently immobilized onto silica fused capillaries (ICERs, 15 cm × 0.1 mm ID). The statistical comparison of the ICERs screening assay with that of the free enzymes is reported and highlights the advantages of the on-flow ICERs assay. Two out of 20 coumarin derivatives could be highlighted: compound 17 is more active toward BChE (IC₅₀=109 ± 21 μM) and 19 showed activity against both enzymes (BChE IC₅₀=128 ± 28 μM and hu-AChE IC₅₀=144 ± 40 μM). The statistical evaluation of the results of the ICERs and free enzyme assays showed no difference between them, further validating the ICERs assay model. The ICERs ability to recognize selective ligands and its use for characterization of the inhibition mechanisms of the hits consolidates the approach here reported.
authors: Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, 14040-901, Ribeirão Preto, SP, Brazil.
authors: Adriana Ferreira Lopes
authors: AF
authors: Vilela
authors: Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, 14040-901, Ribeirão Preto, SP, Brazil.
authors: Joyce Izidoro
authors: JI
authors: da Silva
authors: Departamento de Química, Universidade Federal de São Carlos, Cx. Postal 676, São Carlos, 13565-905, SP, Brazil.
authors: Lucas Campos Curcino
authors: LC
authors: Vieira
authors: Centro Avançado de Estudos e Pesquisas - CAEP, 13087-567, Campinas, SP, Brazil.
authors: Gilberto C R
authors: GC
authors: Bernasconi
authors: Departamento de Química, Universidade Federal de São Carlos, Cx. Postal 676, São Carlos, 13565-905, SP, Brazil.
authors: Arlene Gonçalves
authors: AG
authors: Corrêa
authors: Departamento de Química, Universidade Federal de São Carlos, Cx. Postal 676, São Carlos, 13565-905, SP, Brazil.
authors: Quezia Bezerra
authors: QB
authors: Cass
authors: Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, 14040-901, Ribeirão Preto, SP, Brazil. Electronic address: ccardoso@ffclrp.usp.br.
authors: Carmen Lúcia
authors: CL
authors: Cardoso
mesh_terms: D000110:Acetylcholinesterase / Q000737:chemistry*; D002091:Butyrylcholinesterase / Q000737:chemistry*; D002196:Capillaries / Q000737:chemistry*; D002800:Cholinesterase Inhibitors / Q000737:chemistry*; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry*; D008024:Ligands; D012822:Silicon Dioxide / Q000737:chemistry
chemical_list: D002800:Cholinesterase Inhibitors; D055767:Immobilized Proteins; D008024:Ligands; D012822:Silicon Dioxide; D000110:Acetylcholinesterase; D002091:Butyrylcholinesterase
keywords: Acetylcholinesterase; Butyrylcholinesterase; Enzymatic inhibition assays; Screening; Selective inhibitor; Statistical comparation