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In silico design and biological evaluation of a dual specificity kinase inhibitor targeting cell cycle progression and angiogenesis.

Latham AM, Kankanala J, Fearnley GW, Gage MC, Kearney MT, Homer-Vanniasinkam S, Wheatcroft SB, Fishwick CW, Ponnambalam S - PLoS ONE (2014)

Bottom Line: However, balancing drug specificity and efficacy is problematic with off-target effects and toxicity issues.We deduce that JK-31 reduces the growth of both human endothelial cells and human breast cancer cells in vitro.In silico design is an attractive and innovative method to aid such drug discovery.

View Article: PubMed Central - PubMed

Affiliation: Endothelial Cell Biology Unit, School of Molecular & Cellular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom.

ABSTRACT

Background: Protein kinases play a central role in tumor progression, regulating fundamental processes such as angiogenesis, proliferation and metastasis. Such enzymes are an increasingly important class of drug target with small molecule kinase inhibitors being a major focus in drug development. However, balancing drug specificity and efficacy is problematic with off-target effects and toxicity issues.

Methodology: We have utilized a rational in silico-based approach to demonstrate the design and study of a novel compound that acts as a dual inhibitor of vascular endothelial growth factor receptor 2 (VEGFR2) and cyclin-dependent kinase 1 (CDK1). This compound acts by simultaneously inhibiting pro-angiogenic signal transduction and cell cycle progression in primary endothelial cells. JK-31 displays potent in vitro activity against recombinant VEGFR2 and CDK1/cyclin B proteins comparable to previously characterized inhibitors. Dual inhibition of the vascular endothelial growth factor A (VEGF-A)-mediated signaling response and CDK1-mediated mitotic entry elicits anti-angiogenic activity both in an endothelial-fibroblast co-culture model and a murine ex vivo model of angiogenesis.

Conclusions: We deduce that JK-31 reduces the growth of both human endothelial cells and human breast cancer cells in vitro. This novel synthetic molecule has broad implications for development of similar multi-kinase inhibitors with anti-angiogenic and anti-cancer properties. In silico design is an attractive and innovative method to aid such drug discovery.

No MeSH data available.


Related in: MedlinePlus

JK-31 inhibits endothelial wound closure in vitro.(A) Scratch-wounded endothelial cell monolayers were incubated in either full growth medium (grey bars) or stimulated with 25 ng/ml VEGF-A (black bars) for 16 h in the presence of DMSO or JK-31 (1, 10 or 50 µM). Wounded cell monolayers were photographed before and after treatment and wound widths at each time point were measured using Image J software. Percentage wound closure was calculated by [(width before – width after)/width before] ×100. Error bars represent ± SEM (n = 3; *p<0.05; **p<0.01) for each condition. (B) Representative phase-contrast micrographs depicting wound closure over 16 h in the presence VEGF-A and either DMSO or JK-31 (10 µM).
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pone-0110997-g004: JK-31 inhibits endothelial wound closure in vitro.(A) Scratch-wounded endothelial cell monolayers were incubated in either full growth medium (grey bars) or stimulated with 25 ng/ml VEGF-A (black bars) for 16 h in the presence of DMSO or JK-31 (1, 10 or 50 µM). Wounded cell monolayers were photographed before and after treatment and wound widths at each time point were measured using Image J software. Percentage wound closure was calculated by [(width before – width after)/width before] ×100. Error bars represent ± SEM (n = 3; *p<0.05; **p<0.01) for each condition. (B) Representative phase-contrast micrographs depicting wound closure over 16 h in the presence VEGF-A and either DMSO or JK-31 (10 µM).

Mentions: Endothelial cells in culture form a confluent cell monolayer [16]. A simple in vitro assay can be used to disrupt this cell monolayer, for example by a mechanical scratch wound, and the migration and proliferation of endothelial cells into the denuded region can be monitored over time. This model thus recapitulates early events during angiogenesis [6], [35]. We assessed the effects of JK-31 on endothelial cell monolayer wound closure during growth either in full growth medium or minimal medium supplemented with VEGF-A alone (Figure 4). At 10 µM concentration and above, JK-31 completely inhibited VEGF-A-stimulated wound closure (Figure 4A, 4B) and only partially inhibited wound closure in full growth medium (Figure 4A).


In silico design and biological evaluation of a dual specificity kinase inhibitor targeting cell cycle progression and angiogenesis.

Latham AM, Kankanala J, Fearnley GW, Gage MC, Kearney MT, Homer-Vanniasinkam S, Wheatcroft SB, Fishwick CW, Ponnambalam S - PLoS ONE (2014)

JK-31 inhibits endothelial wound closure in vitro.(A) Scratch-wounded endothelial cell monolayers were incubated in either full growth medium (grey bars) or stimulated with 25 ng/ml VEGF-A (black bars) for 16 h in the presence of DMSO or JK-31 (1, 10 or 50 µM). Wounded cell monolayers were photographed before and after treatment and wound widths at each time point were measured using Image J software. Percentage wound closure was calculated by [(width before – width after)/width before] ×100. Error bars represent ± SEM (n = 3; *p<0.05; **p<0.01) for each condition. (B) Representative phase-contrast micrographs depicting wound closure over 16 h in the presence VEGF-A and either DMSO or JK-31 (10 µM).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4230991&req=5

pone-0110997-g004: JK-31 inhibits endothelial wound closure in vitro.(A) Scratch-wounded endothelial cell monolayers were incubated in either full growth medium (grey bars) or stimulated with 25 ng/ml VEGF-A (black bars) for 16 h in the presence of DMSO or JK-31 (1, 10 or 50 µM). Wounded cell monolayers were photographed before and after treatment and wound widths at each time point were measured using Image J software. Percentage wound closure was calculated by [(width before – width after)/width before] ×100. Error bars represent ± SEM (n = 3; *p<0.05; **p<0.01) for each condition. (B) Representative phase-contrast micrographs depicting wound closure over 16 h in the presence VEGF-A and either DMSO or JK-31 (10 µM).
Mentions: Endothelial cells in culture form a confluent cell monolayer [16]. A simple in vitro assay can be used to disrupt this cell monolayer, for example by a mechanical scratch wound, and the migration and proliferation of endothelial cells into the denuded region can be monitored over time. This model thus recapitulates early events during angiogenesis [6], [35]. We assessed the effects of JK-31 on endothelial cell monolayer wound closure during growth either in full growth medium or minimal medium supplemented with VEGF-A alone (Figure 4). At 10 µM concentration and above, JK-31 completely inhibited VEGF-A-stimulated wound closure (Figure 4A, 4B) and only partially inhibited wound closure in full growth medium (Figure 4A).

Bottom Line: However, balancing drug specificity and efficacy is problematic with off-target effects and toxicity issues.We deduce that JK-31 reduces the growth of both human endothelial cells and human breast cancer cells in vitro.In silico design is an attractive and innovative method to aid such drug discovery.

View Article: PubMed Central - PubMed

Affiliation: Endothelial Cell Biology Unit, School of Molecular & Cellular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom.

ABSTRACT

Background: Protein kinases play a central role in tumor progression, regulating fundamental processes such as angiogenesis, proliferation and metastasis. Such enzymes are an increasingly important class of drug target with small molecule kinase inhibitors being a major focus in drug development. However, balancing drug specificity and efficacy is problematic with off-target effects and toxicity issues.

Methodology: We have utilized a rational in silico-based approach to demonstrate the design and study of a novel compound that acts as a dual inhibitor of vascular endothelial growth factor receptor 2 (VEGFR2) and cyclin-dependent kinase 1 (CDK1). This compound acts by simultaneously inhibiting pro-angiogenic signal transduction and cell cycle progression in primary endothelial cells. JK-31 displays potent in vitro activity against recombinant VEGFR2 and CDK1/cyclin B proteins comparable to previously characterized inhibitors. Dual inhibition of the vascular endothelial growth factor A (VEGF-A)-mediated signaling response and CDK1-mediated mitotic entry elicits anti-angiogenic activity both in an endothelial-fibroblast co-culture model and a murine ex vivo model of angiogenesis.

Conclusions: We deduce that JK-31 reduces the growth of both human endothelial cells and human breast cancer cells in vitro. This novel synthetic molecule has broad implications for development of similar multi-kinase inhibitors with anti-angiogenic and anti-cancer properties. In silico design is an attractive and innovative method to aid such drug discovery.

No MeSH data available.


Related in: MedlinePlus