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Structure-Activity Relationship of Indole-Tethered Pyrimidine Derivatives that Concurrently Inhibit Epidermal Growth Factor Receptor and Other Angiokinases.

Song J, Yoo J, Kwon A, Kim D, Nguyen HK, Lee BY, Suh W, Min KH - PLoS ONE (2015)

Bottom Line: MKP101 inhibited not only the epidermal growth factor receptor with an IC50 value of 43 nM but also inhibited angiokinases as potently as pazopanib.A docking model of MKP101 and the kinase domain of the epidermal growth factor receptor was generated to predict its binding mode, and validated by synthesizing and evaluating MKP101 derivatives.We believe that this study could provide a basis for developing angiokinase inhibitors having high affinity for the epidermal growth factor receptor, from the pyrimidine scaffold.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Chung-Ang University, Seoul, Republic of Korea.

ABSTRACT
Antiangiogenic agents have been widely investigated in combination with standard chemotherapy or targeted cancer agents for better management of advanced cancers. Therapeutic agents that concurrently inhibit epidermal growth factor receptor and other angiokinases could be useful alternatives to combination therapies for epidermal growth factor receptor-dependent cancers. Here, we report the synthesis of an indole derivative of pazopanib using a bioisosteric replacement strategy, which was designated MKP101. MKP101 inhibited not only the epidermal growth factor receptor with an IC50 value of 43 nM but also inhibited angiokinases as potently as pazopanib. In addition, MKP101 effectively inhibited vascular endothelial growth factor-induced endothelial proliferation, tube formation, migration of human umbilical vein endothelial cells and proliferation of HCC827, an epidermal growth factor receptor-addicted cancer cell line. A docking model of MKP101 and the kinase domain of the epidermal growth factor receptor was generated to predict its binding mode, and validated by synthesizing and evaluating MKP101 derivatives. Additionally, a study of structure-activity relationships of indolylamino or indolyloxy pyrimidine analogues derived from MKP101 demonstrated that selectivity for epidermal growth factor receptor and other angiokinases, especially vascular endothelial growth factor receptor 2 depends on the position of substituents on pyrimidine and the type of link between pyrimidine and the indole moiety. We believe that this study could provide a basis for developing angiokinase inhibitors having high affinity for the epidermal growth factor receptor, from the pyrimidine scaffold.

No MeSH data available.


Related in: MedlinePlus

MKP101 inhibits proliferation of epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor (TKI)-sensitive non-small cell lung cancer (NSCLC) cells.(A) Inhibition of HCC827 cell proliferation. Cells were incubated for 72 h with a range of concentrations of MKP101, and cell viability was measured using the WST-1 assay. The IC50 value was 160 nM. (B) Inhibition of EGFR phosphorylation in HCC827 cells. Western blot analysis using antibodies to phosphorylated and total EGFR from a representative experiment is shown. Cells were incubated for 3 h at the indicated concentration. (C) Graph showing the relative intensities of the total and phosphorylated EGFR as determined by band densitometry. IC50, half-maximal inhibitory concentration.
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pone.0138823.g003: MKP101 inhibits proliferation of epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor (TKI)-sensitive non-small cell lung cancer (NSCLC) cells.(A) Inhibition of HCC827 cell proliferation. Cells were incubated for 72 h with a range of concentrations of MKP101, and cell viability was measured using the WST-1 assay. The IC50 value was 160 nM. (B) Inhibition of EGFR phosphorylation in HCC827 cells. Western blot analysis using antibodies to phosphorylated and total EGFR from a representative experiment is shown. Cells were incubated for 3 h at the indicated concentration. (C) Graph showing the relative intensities of the total and phosphorylated EGFR as determined by band densitometry. IC50, half-maximal inhibitory concentration.

Mentions: Derivatives of pazopanib, primarily those with benzenesulfonamido moieties have been reported previously [27,28,29]. Some benzenesulfonamido pazopanib derivatives inhibited VEGF more potently than pazopanib, but did not exhibit considerable activity for EGFR [28]. Thus our derivatization strategy focused on the indazole moiety of pazopanib. The indole moiety is a viable bioisostere of indazole and is, therefore, suitable for the replacement strategy, [30]. Therefore, an indole moiety was introduced in the pazopanib molecule, and the resulting compound MKP101 was evaluated for effects on EGFR and angiokinases (Fig 2). MKP101 strongly inhibited wild type EGFR and the mutant EGFR L858R with IC50 values of 43 and 17 nM, respectively, while pazopanib did not inhibit EGFR. MKP101 also significantly inhibited angiokinases including VEGFRs, FGFR3, PDGFRs, and cKit (Table 1). Kumar et al. provided IC50 values of pazopanib against 61 kinases [31], which were compared to the kinase profile data of MKP101. The potency of inhibition of the angiokinases by MKP101 was similar to that induced by pazopanib in the in vitro kinase assay. In addition, 40 kinases were profiled to determine the selectivity of MKP101. Of the 40 kinases tested, only 3 kinases were inhibited by more than 90% by 1 μM MKP101 (Table 2). Furthermore, MKP101 was evaluated for anti-proliferative activity against HCC827 cells, an EGFR TKI-sensitive NSCLC cell line. In HCC827 cells, MKP101 significantly inhibited proliferation with an IC50 value of 160 nM (Fig 3). Gefitinib was tested as a positive control, and had an IC50 value of 10 nM. The inhibition of EGFR phosphorylation by MKP101 was demonstrated by western blot analysis, in which MKP101 reduced the levels of phosphorylated EGFR.


Structure-Activity Relationship of Indole-Tethered Pyrimidine Derivatives that Concurrently Inhibit Epidermal Growth Factor Receptor and Other Angiokinases.

Song J, Yoo J, Kwon A, Kim D, Nguyen HK, Lee BY, Suh W, Min KH - PLoS ONE (2015)

MKP101 inhibits proliferation of epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor (TKI)-sensitive non-small cell lung cancer (NSCLC) cells.(A) Inhibition of HCC827 cell proliferation. Cells were incubated for 72 h with a range of concentrations of MKP101, and cell viability was measured using the WST-1 assay. The IC50 value was 160 nM. (B) Inhibition of EGFR phosphorylation in HCC827 cells. Western blot analysis using antibodies to phosphorylated and total EGFR from a representative experiment is shown. Cells were incubated for 3 h at the indicated concentration. (C) Graph showing the relative intensities of the total and phosphorylated EGFR as determined by band densitometry. IC50, half-maximal inhibitory concentration.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4581874&req=5

pone.0138823.g003: MKP101 inhibits proliferation of epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor (TKI)-sensitive non-small cell lung cancer (NSCLC) cells.(A) Inhibition of HCC827 cell proliferation. Cells were incubated for 72 h with a range of concentrations of MKP101, and cell viability was measured using the WST-1 assay. The IC50 value was 160 nM. (B) Inhibition of EGFR phosphorylation in HCC827 cells. Western blot analysis using antibodies to phosphorylated and total EGFR from a representative experiment is shown. Cells were incubated for 3 h at the indicated concentration. (C) Graph showing the relative intensities of the total and phosphorylated EGFR as determined by band densitometry. IC50, half-maximal inhibitory concentration.
Mentions: Derivatives of pazopanib, primarily those with benzenesulfonamido moieties have been reported previously [27,28,29]. Some benzenesulfonamido pazopanib derivatives inhibited VEGF more potently than pazopanib, but did not exhibit considerable activity for EGFR [28]. Thus our derivatization strategy focused on the indazole moiety of pazopanib. The indole moiety is a viable bioisostere of indazole and is, therefore, suitable for the replacement strategy, [30]. Therefore, an indole moiety was introduced in the pazopanib molecule, and the resulting compound MKP101 was evaluated for effects on EGFR and angiokinases (Fig 2). MKP101 strongly inhibited wild type EGFR and the mutant EGFR L858R with IC50 values of 43 and 17 nM, respectively, while pazopanib did not inhibit EGFR. MKP101 also significantly inhibited angiokinases including VEGFRs, FGFR3, PDGFRs, and cKit (Table 1). Kumar et al. provided IC50 values of pazopanib against 61 kinases [31], which were compared to the kinase profile data of MKP101. The potency of inhibition of the angiokinases by MKP101 was similar to that induced by pazopanib in the in vitro kinase assay. In addition, 40 kinases were profiled to determine the selectivity of MKP101. Of the 40 kinases tested, only 3 kinases were inhibited by more than 90% by 1 μM MKP101 (Table 2). Furthermore, MKP101 was evaluated for anti-proliferative activity against HCC827 cells, an EGFR TKI-sensitive NSCLC cell line. In HCC827 cells, MKP101 significantly inhibited proliferation with an IC50 value of 160 nM (Fig 3). Gefitinib was tested as a positive control, and had an IC50 value of 10 nM. The inhibition of EGFR phosphorylation by MKP101 was demonstrated by western blot analysis, in which MKP101 reduced the levels of phosphorylated EGFR.

Bottom Line: MKP101 inhibited not only the epidermal growth factor receptor with an IC50 value of 43 nM but also inhibited angiokinases as potently as pazopanib.A docking model of MKP101 and the kinase domain of the epidermal growth factor receptor was generated to predict its binding mode, and validated by synthesizing and evaluating MKP101 derivatives.We believe that this study could provide a basis for developing angiokinase inhibitors having high affinity for the epidermal growth factor receptor, from the pyrimidine scaffold.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Chung-Ang University, Seoul, Republic of Korea.

ABSTRACT
Antiangiogenic agents have been widely investigated in combination with standard chemotherapy or targeted cancer agents for better management of advanced cancers. Therapeutic agents that concurrently inhibit epidermal growth factor receptor and other angiokinases could be useful alternatives to combination therapies for epidermal growth factor receptor-dependent cancers. Here, we report the synthesis of an indole derivative of pazopanib using a bioisosteric replacement strategy, which was designated MKP101. MKP101 inhibited not only the epidermal growth factor receptor with an IC50 value of 43 nM but also inhibited angiokinases as potently as pazopanib. In addition, MKP101 effectively inhibited vascular endothelial growth factor-induced endothelial proliferation, tube formation, migration of human umbilical vein endothelial cells and proliferation of HCC827, an epidermal growth factor receptor-addicted cancer cell line. A docking model of MKP101 and the kinase domain of the epidermal growth factor receptor was generated to predict its binding mode, and validated by synthesizing and evaluating MKP101 derivatives. Additionally, a study of structure-activity relationships of indolylamino or indolyloxy pyrimidine analogues derived from MKP101 demonstrated that selectivity for epidermal growth factor receptor and other angiokinases, especially vascular endothelial growth factor receptor 2 depends on the position of substituents on pyrimidine and the type of link between pyrimidine and the indole moiety. We believe that this study could provide a basis for developing angiokinase inhibitors having high affinity for the epidermal growth factor receptor, from the pyrimidine scaffold.

No MeSH data available.


Related in: MedlinePlus