Made available in DSpace on 2022-04-29T08:35:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) Falcipain-2 (FP-2) is a Plasmodium falciparum hemoglobinase widely targeted in the search for antimalarials. FP-2 can be allosterically modulated by various noncompetitive inhibitors that have been serendipitously identified. Moreover, the crystal structures of two inhibitors bound to an allosteric site, termed site 6, of the homolog enzyme human cathepsin K (hCatK) suggest that the equivalent region in FP-2 might play a similar role. Here, we conduct the rational identification of FP-2 inhibitors through virtual screenings (VS) of compounds into several pocket-like conformations of site 6, sampled during molecular dynamics (MD) simulations of the free enzyme. Two noncompetitive inhibitors, ZINC03225317 and ZINC72290660, were confirmed using in vitro enzymatic assays and their poses into site 6 led to calculated binding free energies matching the experimental ones. Our results provide strong evidence about the allosteric inhibition of FP-2 through binding of small molecules to site 6, thus opening the way toward the discovery of new inhibitors against this enzyme. Departamento de Física Instituto de Biociências Letras e Ciências Exatas – Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Rua Cristóvão Colombo 2265, Jardim Nazareth Laboratório de Modelagem e Dinâmica Molecular Instituto de Biofı́sica Carlos Chagas Filho Universidade Federal do Rio de Janeiro, Ave. Carlos Chagas Filho – Universidade Federal do Rio de Janeiro (UFRJ), Ave. Carlos Chagas Filho, 373, CCS-Bloco D sala 30, Cidade Universitária Ilha de Fundão Institute for Molecular Modeling and Simulation Department for Material Sciences and Process Engineering – University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18 Instituto de Investigaciones Biotecnológicas Dr. Rodolfo Ugalde Universidad Nacional de San Martín CONICET, San Martín Instituto de Ciências Exatas Universidade Federal de Juiz de Fora (UFJF), Rua José Lourenço Kelmer, s/n – Campus Universitário, Bairro São Pedro Departamento de Física Instituto de Biociências Letras e Ciências Exatas – Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Rua Cristóvão Colombo 2265, Jardim Nazareth