Potential Antileukemia Effect and Structural Analyses of SRPK Inhibition by N-(2-(Piperidin-1-yl)-5-(Trifluoromethyl)Phenyl)Isonicotinamide (SRPIN340)

Raoni Pais Siqueira; Éverton de Almeida Alves Barbosa; Marcelo Depólo Polêto; Germanna Lima Righetto; Thiago Vargas Seraphim; Rafael Locatelli Salgado; Joana Gasperazzo Ferreira; Marcus Vinícius de Andrade Barros; Leandro Licursi de Oliveira; Angelo Brunelli Albertoni Laranjeira; Márcia Rogéria Almeida; Abelardo Silva Júnior; Juliana Lopes Rangel Fietto; Jörg Kobarg; Eduardo Basílio de Oliveira; Robson Ricardo Teixeira; Júlio César Borges; Jose Andrés Yunes; Gustavo Costa Bressan

doi:10.1371/journal.pone.0134882

Potential Antileukemia Effect and Structural Analyses of SRPK Inhibition by N-(2-(Piperidin-1-yl)-5-(Trifluoromethyl)Phenyl)Isonicotinamide (SRPIN340)

PLoS One. 2015 Aug 5;10(8):e0134882. doi: 10.1371/journal.pone.0134882. eCollection 2015.

Authors

Raoni Pais Siqueira¹, Éverton de Almeida Alves Barbosa¹, Marcelo Depólo Polêto¹, Germanna Lima Righetto², Thiago Vargas Seraphim³, Rafael Locatelli Salgado¹, Joana Gasperazzo Ferreira¹, Marcus Vinícius de Andrade Barros⁴, Leandro Licursi de Oliveira⁵, Angelo Brunelli Albertoni Laranjeira⁶, Márcia Rogéria Almeida¹, Abelardo Silva Júnior⁷, Juliana Lopes Rangel Fietto¹, Jörg Kobarg⁸, Eduardo Basílio de Oliveira⁹, Robson Ricardo Teixeira⁴, Júlio César Borges³, Jose Andrés Yunes⁶, Gustavo Costa Bressan¹

Affiliations

¹ Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brasil.
² Instituto de Biologia, Universidade Estadual de Campinas, Campinas, São Paulo, Brasil.
³ Instituto de Química, Universidade de São Paulo, São Carlos, São Paulo, Brasil.
⁴ Departamento de Química, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brasil.
⁵ Departamento de Biologia Geral, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brasil.
⁶ Centro Infantil Boldrini, Campinas, São Paulo, Brasil.
⁷ Departamento de Veterinária, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brasil.
⁸ Laboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e Materiais, Campinas, São Paulo, Brasil.
⁹ Departamento de Engenharia de Alimentos, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brasil.

Abstract

Dysregulation of pre-mRNA splicing machinery activity has been related to the biogenesis of several diseases. The serine/arginine-rich protein kinase family (SRPKs) plays a critical role in regulating pre-mRNA splicing events through the extensive phosphorylation of splicing factors from the family of serine/arginine-rich proteins (SR proteins). Previous investigations have described the overexpression of SRPK1 and SRPK2 in leukemia and other cancer types, suggesting that they would be useful targets for developing novel antitumor strategies. Herein, we evaluated the effect of selective pharmacological SRPK inhibition by N-(2-(piperidin-1-yl)-5-(trifluoromethyl)phenyl)isonicotinamide (SRPIN340) on the viability of lymphoid and myeloid leukemia cell lines. Along with significant cytotoxic activity, the effect of treatments in regulating the phosphorylation of the SR protein family and in altering the expression of MAP2K1, MAP2K2, VEGF and FAS genes were also assessed. Furthermore, we found that pharmacological inhibition of SRPKs can trigger early and late events of apoptosis. Finally, intrinsic tryptophan fluorescence emission, molecular docking and molecular dynamics were analyzed to gain structural information on the SRPK/SRPIN340 complex. These data suggest that SRPK pharmacological inhibition should be considered as an alternative therapeutic strategy for fighting leukemias. Moreover, the obtained SRPK-ligand interaction data provide useful structural information to guide further medicinal chemistry efforts towards the development of novel drug candidates.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antineoplastic Agents / chemistry
Antineoplastic Agents / metabolism
Antineoplastic Agents / pharmacology*
Apoptosis / drug effects*
Binding Sites
Blotting, Western
Cell Survival / drug effects
Cells, Cultured
Gene Expression Regulation, Leukemic
HL-60 Cells
HeLa Cells
Humans
Jurkat Cells
K562 Cells
Leukemia / genetics
Leukemia / metabolism
Leukemia / pathology
Molecular Docking Simulation
Molecular Dynamics Simulation
Molecular Structure
Niacinamide / analogs & derivatives*
Niacinamide / chemistry
Niacinamide / metabolism
Niacinamide / pharmacology
Piperidines / chemistry
Piperidines / metabolism
Piperidines / pharmacology*
Protein Binding
Protein Serine-Threonine Kinases / antagonists & inhibitors*
Protein Serine-Threonine Kinases / chemistry
Protein Serine-Threonine Kinases / metabolism
Protein Structure, Tertiary
Reverse Transcriptase Polymerase Chain Reaction
Spectrometry, Fluorescence

Substances

Antineoplastic Agents
Piperidines
SRPIN340
Niacinamide
SRPK1 protein, human
Protein Serine-Threonine Kinases
SRPK2 protein, human

Grants and funding

This study was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico, http://www.cnpq.br/ (CNPq; grant 485011/2012-3 to GCB); Fundação de Amparo à Pesquisa do Estado de Minas Gerais, http://www.fapemig.br/ (FAPEMIG; grant CBB-APQ-01637-13 to GCB); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, http://www.capes.gov.br/ (CAPES; grant CAPES/CNPq 552459/2011-9 to JLRF); Fundação Arthur Bernardes, www.funarbe.org.br/ (FUNARBE; grant FUNARPEX/2014 to GCB). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.