Application of capillary bioreactor of acetylcholinesterase in screening of combining collections and natural crude extracts

• Grant number: 12/01800-8
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: April 01, 2012
• End date: March 31, 2013
• Field of knowledge: Physical Sciences and Mathematics - Chemistry - Organic Chemistry
• Principal Investigator: Carmen Lúcia Cardoso
• Grantee: Bárbara Mammana Frugeri
• Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 08/04371-5 - Development and applications of enzymatic columns in selective inhibitors screening and in the natural products studies, AP.JP

Abstract

There is great interest in the development of fast and effective techniques for the screening of substances in drug discovery research. The improvement of techniques, along with the development of high-efficiency tests, has allowed for the utilization of enzymes, receptors, as well as other targets in the evaluation of samples and in the identification of possible drug candidates that bind to biological targets. One interesting method consists of the immobilization of a biomolecule (for example, enzymes) to chromatographic supports, which results in a bioreactor. In this context, enzymatic bioreactors using acetylcholinesterase (AChE) from human erythrocytes and an identification method of selective inhibitors have been developed and validated. The resulting method has been employed in the screening of collections of synthetic compounds planned as AChE inhibitors and of natural extracts in order to identify the substances with better activities. Studies on IC50 and action mechanisms will be performed on the compounds with better action, and their selectivity will be determined by comparison of the inhibitory activity displayed in the bioreactors containing different cholinesterase enzymes, namelly AChE from Electrophorus electricus and human erythrocytes and BuChE (butyrylcholinesterase from human erythrocytes.(AU)