Pyrrole-indolinone SU11652 targets the nucleoside diphosphate kinase from Leishmania parasites

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Vieira, Plinio Salmazo ; Campos Brasil Souza, Tatiana de Arruda ; Honorato, Rodrigo Vargas ; Zanphorlin, Leticia Maria ; Severiano, Kelven Ulisses ; Rocco, Silvana Aparecida ; Cavalcante de Oliveira, Arthur Henrique ; Cordeiro, Artur Torres ; Lopes Oliveira, Paulo Sergio ; de Giuseppe, Priscila Oliveira ; Murakami, Mario Tyago

• Document type: Journal article
• Source: Biochemical and Biophysical Research Communications; v. 488, n. 3, p. 461-465, JUL 1 2017.
• Web of Science Citations: 3

Abstract

Nucleoside diphosphate kinases (NDKs) are key enzymes in the purine-salvage pathway of trypanosomatids and have been associated with the maintenance of host-cell integrity for the benefit of the parasite, being potential targets for rational drug discovery and design. The NDK from Leishmania major (LmNDK) and mutants were expressed and purified to homogeneity. Thermal shift assays were employed to identify potential inhibitors for LmNDK. Calorimetric experiments, site-directed mutagenesis and molecular docking analysis were performed to validate the interaction and to evaluate the structural basis of ligand recognition. Furthermore, the anti-leishmanial activity of the newly identified and validated compound was tested in vitro against different Leishmania species. The molecule SU11652, a Sunitinib analog, was identified as a potential inhibitor for LmNDK and structural studies indicated that this molecule binds to the active site of LmNDK in a similar conformation to nucleotides, mimicking natural substrates. Isothermal titration calorimetry experiments combined with site-directed mutagenesis revealed that the residues H50 and H117, considered essential for catalysis, play an important role in ligand binding. In vitro cell studies showed that SU11652 had similar efficacy to Amphotericin b against some Leishmania species. Together, our results indicate the pyrrole-indolinone SU11652 as a promising scaffold for the rational design of new drugs targeting the enzyme NDK from Leishmania parasites. (C) 2017 Elsevier Inc. All rights reserved. (AU)

• FAPESP's process: 11/20569-2 - Role of amino acids of the active site and interface in oligomeric in the structure and function of the nucleoside diphosphate kinase of Leishmania major
• Grantee: Arthur Henrique Cavalcante de Oliveira
• Support Opportunities: Regular Research Grants
• FAPESP's process: 10/03761-4 - Kinetic characterization and effect of mutations in the oligomeric interface of the Nucleoside Diphosphate Kinase from Leishmania major.
• Grantee: Plínio Salmazo Vieira
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 07/06755-2 - Structural and functional characterization of proteins envolved in the virulence of the protozoan parasite Leishmania major
• Grantee: Arthur Henrique Cavalcante de Oliveira
• Support Opportunities: Regular Research Grants
• FAPESP's process: 11/24178-8 - Structural studies of NEK kinases from Leishmania braziliensis.
• Grantee: Plínio Salmazo Vieira
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 10/51730-0 - Functional and structural studies of protein kinases involved in cancer and neglected diseases: towards the development of new inhibitors
• Grantee: Jörg Kobarg
• Support Opportunities: Regular Research Grants