Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/244039
Type: Artigo de periódico
Title: Is Rk-682 A Promiscuous Enzyme Inhibitor? Synthesis And In Vitro Evaluation Of Protein Tyrosine Phosphatase Inhibition Of Racemic Rk-682 And Analogues
Author: Carneiro
Vania M. T.; Trivella
Daniela B. B.; Scorsato
Valeria; Beraldo
Viviane L.; Dias
Mariana P.; Sobreira
Tiago J. P.; Aparicio
Ricardo; Pilli
Ronaldo A.
Abstract: RK-682 (1) is a natural product known to selectively inhibit protein tyrosine phosphatases (PTPases) and is used commercially as a positive control for phosphatase inhibition in in vitro assays. Protein phosphatases are involved in several human diseases including diabetes, cancer and inflammation, and are considered important targets for pharmaceutical development. Here we report the synthesis of racemic RK-682 (rac-1) and a focused set of compounds, including racemic analogues of I, dihydropyranones and C-acylated Meldrum's acid derivatives, the later obtained in one synthetic step from commercially available starting material. We further characterized the behavior of some representative compounds in aqueous solution and evaluated their in vitro PTPase binding and inhibition. Our data reveal that rac-1 and some derivatives are able to form large aggregates in solution, in which the aggregation capacity is dependent on the acyl side chain size. However, compound aggregation per se is not able to promote PTPase inhibition. Our data disclose a novel family of PTPase inhibitors (C-acylated Meldrum's acid derivatives) and that rac-1 and derivatives with an exposed latent negatively charged substructure (e.g.: the tetronic acid core of 1) can bind to the PTPase binding site, as well promiscuously to protein surfaces. The combined capacity of compounds to bind to proteins together with their intrinsic capacity to aggregate in solution seems essential to promote enzyme aggregation and thus, its inhibition. We also observed that divalent cations, such as magnesium frequently used in enzyme buffer solutions, can deplete the inhibitory activity of rac-1, thus influencing the enzyme inhibition experiment. Overall, these data help to characterize the mechanism of PTPase inhibition by rac-1 and derivatives, revealing that enzyme inhibition is not solely dependent on compound binding to the PTPase catalytic site as generally accepted in the literature. In addition, our results point to promiscuous mechanisms that influence significantly the in vitro evaluation of enzyme inhibition by rac-1. Therefore, we recommend caution when using natural or synthetic RK-682 (1) as an internal control for evaluating PTPase inhibition and selectivity, since many events can modulate the apparent enzyme inhibition. (C) 2015 Elsevier Masson SAS. All rights reserved.
Subject: Dual-specificity Phosphatase
Tetronic Acid Homologs
Long-term Potentiation
Extracellular Phosphorylation
Asymmetric-synthesis
Hiv-1 Protease
Ca1 Neurons
Cell-cycle
Mechanism
Cdc25b
Country: PARIS
Editor: ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER
Citation: Is Rk-682 A Promiscuous Enzyme Inhibitor? Synthesis And In Vitro Evaluation Of Protein Tyrosine Phosphatase Inhibition Of Racemic Rk-682 And Analogues. Elsevier France-editions Scientifiques Medicales Elsevier, v. 97, p. 42-54 Jun-2015.
Rights: embargo
Identifier DOI: 10.1016/j.ejmech.2015.04.036
Address: http://www.sciencedirect.com/science/article/pii/S0223523415002950
Date Issue: 2015
Appears in Collections:Unicamp - Artigos e Outros Documentos

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