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

Siqueira RP, Barbosa Éde A, Polêto MD, Righetto GL, Seraphim TV, Salgado RL, Ferreira JG, Barros MV, de Oliveira LL, Laranjeira AB, Almeida MR, Júnior AS, Fietto JL, Kobarg J, de Oliveira EB, Teixeira RR, Borges JC, Yunes JA, Bressan GC - PLoS ONE (2015)

Bottom Line: 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.Moreover, the obtained SRPK-ligand interaction data provide useful structural information to guide further medicinal chemistry efforts towards the development of novel drug candidates.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Bioquímica e Biologia Molecular, 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.

No MeSH data available.


Related in: MedlinePlus

The effect of SRPIN340 treatment on leukemia cell viability and death.Different leukemia cell lines (A) were treated with increasing concentrations (0–100 μM) of SRPIN340 for 48 h. Cell viability was determined using the MTT assay. We considered the viability of 100% of the cells in the control treatment (vehicle). The percentage of inhibition was calculated relative to cells treated with the vehicle. The values are expressed as the means ± standard deviation of three independent experiments. To assess cell death (B), Jurkat cells were treated with 25 μM of SRPIN340 for 6 or 12 h. Cells treated with the vehicle were used as controls. Subsequently, the cell death was evaluated using annexin V-FITC (V) and PI (P) labels. One representative experiment of three is shown.
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pone.0134882.g002: The effect of SRPIN340 treatment on leukemia cell viability and death.Different leukemia cell lines (A) were treated with increasing concentrations (0–100 μM) of SRPIN340 for 48 h. Cell viability was determined using the MTT assay. We considered the viability of 100% of the cells in the control treatment (vehicle). The percentage of inhibition was calculated relative to cells treated with the vehicle. The values are expressed as the means ± standard deviation of three independent experiments. To assess cell death (B), Jurkat cells were treated with 25 μM of SRPIN340 for 6 or 12 h. Cells treated with the vehicle were used as controls. Subsequently, the cell death was evaluated using annexin V-FITC (V) and PI (P) labels. One representative experiment of three is shown.

Mentions: The cytotoxic potential of SRPIN340 was evaluated against CML, AML, ALL-B and ALL-T cell lineages. All evaluated cell lines were sensitive to the treatments, indicating that SRPK inhibition indeed yields an overall antileukemia effect in vitro (Fig 2A). Half-maximal inhibitory concentration values (IC50) (Table 1) were determined and revealed that, in general, myeloid leukemias were more sensitive than lymphoid ones. In this case, the AML HL60 was the most sensitive (IC50 of 44.7 μM) compared with ALL-T Molt4 and Jurkat (IC50 values of 92.2 μM and 82.3 μM, respectively). Moreover, although an accurate IC50 value could not be determined at higher compound concentrations, PBMC seemed to be less sensitive to SRPIN340 than the leukemia lineages evaluated. This suggests that SRPK pharmacological inhibition can selectively affect tumor cell survival.


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

Siqueira RP, Barbosa Éde A, Polêto MD, Righetto GL, Seraphim TV, Salgado RL, Ferreira JG, Barros MV, de Oliveira LL, Laranjeira AB, Almeida MR, Júnior AS, Fietto JL, Kobarg J, de Oliveira EB, Teixeira RR, Borges JC, Yunes JA, Bressan GC - PLoS ONE (2015)

The effect of SRPIN340 treatment on leukemia cell viability and death.Different leukemia cell lines (A) were treated with increasing concentrations (0–100 μM) of SRPIN340 for 48 h. Cell viability was determined using the MTT assay. We considered the viability of 100% of the cells in the control treatment (vehicle). The percentage of inhibition was calculated relative to cells treated with the vehicle. The values are expressed as the means ± standard deviation of three independent experiments. To assess cell death (B), Jurkat cells were treated with 25 μM of SRPIN340 for 6 or 12 h. Cells treated with the vehicle were used as controls. Subsequently, the cell death was evaluated using annexin V-FITC (V) and PI (P) labels. One representative experiment of three is shown.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4526641&req=5

pone.0134882.g002: The effect of SRPIN340 treatment on leukemia cell viability and death.Different leukemia cell lines (A) were treated with increasing concentrations (0–100 μM) of SRPIN340 for 48 h. Cell viability was determined using the MTT assay. We considered the viability of 100% of the cells in the control treatment (vehicle). The percentage of inhibition was calculated relative to cells treated with the vehicle. The values are expressed as the means ± standard deviation of three independent experiments. To assess cell death (B), Jurkat cells were treated with 25 μM of SRPIN340 for 6 or 12 h. Cells treated with the vehicle were used as controls. Subsequently, the cell death was evaluated using annexin V-FITC (V) and PI (P) labels. One representative experiment of three is shown.
Mentions: The cytotoxic potential of SRPIN340 was evaluated against CML, AML, ALL-B and ALL-T cell lineages. All evaluated cell lines were sensitive to the treatments, indicating that SRPK inhibition indeed yields an overall antileukemia effect in vitro (Fig 2A). Half-maximal inhibitory concentration values (IC50) (Table 1) were determined and revealed that, in general, myeloid leukemias were more sensitive than lymphoid ones. In this case, the AML HL60 was the most sensitive (IC50 of 44.7 μM) compared with ALL-T Molt4 and Jurkat (IC50 values of 92.2 μM and 82.3 μM, respectively). Moreover, although an accurate IC50 value could not be determined at higher compound concentrations, PBMC seemed to be less sensitive to SRPIN340 than the leukemia lineages evaluated. This suggests that SRPK pharmacological inhibition can selectively affect tumor cell survival.

Bottom Line: 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.Moreover, the obtained SRPK-ligand interaction data provide useful structural information to guide further medicinal chemistry efforts towards the development of novel drug candidates.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Bioquímica e Biologia Molecular, 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.

No MeSH data available.


Related in: MedlinePlus