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

• Published in: Biochemical and biophysical research communications Vol. 488; no. 3; pp. 461 - 465
• Main Authors: Vieira, Plínio Salmazo, Souza, Tatiana de Arruda Campos Brasil, Honorato, Rodrigo Vargas, Zanphorlin, Letícia Maria, Severiano, Kelven Ulisses, Rocco, Silvana Aparecida, de Oliveira, Arthur Henrique Cavalcante, Cordeiro, Artur Torres, Oliveira, Paulo Sérgio Lopes, de Giuseppe, Priscila Oliveira, Murakami, Mário Tyago
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.2017
• Subjects: Antiprotozoal Agents - pharmacology; Biophysical characterization; Calorimetry; Cell Survival - drug effects; Competitive inhibitor; Dose-Response Relationship, Drug; Indoles - pharmacology; Leishmania; Leishmania major - drug effects; Leishmania major - enzymology; Molecular Docking Simulation; Mutagenesis, Site-Directed; Nucleoside diphosphate kinase; Nucleoside-Diphosphate Kinase - antagonists & inhibitors; Nucleoside-Diphosphate Kinase - genetics; Nucleoside-Diphosphate Kinase - metabolism; Parasitic Sensitivity Tests; Protein Kinase Inhibitors - chemistry; Protein Kinase Inhibitors - pharmacology; Purine-salvage pathway; Pyrrole-indolinone compounds; Pyrroles - pharmacology; Structure-Activity Relationship; Pyrrole-indolinone compounds; NDK; LmNDK; Leishmania; DSF; Purine-salvage pathway; ITC; Tm; Biophysical characterization; Nucleoside diphosphate kinase; Competitive inhibitor; SU11652 (PubChem CID: 5329103)
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2017.05.048

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. [Display omitted] •A Sunitinib variant, SU11652, was identified as a potential inhibitor of L. major NDK.•The ligand mimics natural substrates in the enzyme active site.•The enzyme residues H50 and H117 are important to ligand binding and catalytic activity.•SU11652 has similar efficacy to Amphotericin b against some Leishmania species.