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Journal of Molecular and Clinical Medicine  2018, Vol. 1 Issue (2): 77-84    DOI: 10.31083/j.jmcm.2018.02.004
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Differential mechanism of action of the CK1$\varepsilon $ inhibitor GSD0054
Irene Lagunes1, Elva Martín-Batista1, Gastón Silveira-Dorta1, Miguel X. Fernandes1, 2, José M. Padrón1, *()
1 BioLab, Instituto Universitario de Bio-Orgánica "Antonio González" (IUBO-AG), Centro de Investigaciones Biomédicas de Canarias (CIBICAN), Universidad de La Laguna, C/ Astrofísico Francisco Sánchez 2, 38206 La Laguna, Spain
2 Centro de Química da Madeira, Universidade da Madeira, Campus da Penteada, 9000-390 Funchal, Portugal
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In the current study, we explored for the first time, the mechanism of action of the new Casein kinase 1 $\varepsilon$ (CK1$\varepsilon$) selective inhibitor GSD0054. Although GSD0054 behaved as a selective CK1$\varepsilon $ inhibitor in enzymatic assays, we studied whether this inhibitory activity also occurred inside the cells. The effects of GSD0054 on $\beta $-catenin expression and disruption of cell cycle progression were studied in the human breast cancer cell lines MDA-MB-453 ($\beta $-catenin negative) and T-47D ($\beta $-catenin positive). We also performed molecular modeling studies using computational docking against CK1$\varepsilon $ to explain and predict the mechanism of action of this compound. Moreover, the commercially available CK1$\varepsilon $ inhibitor PF-4800567 and the CK1$\delta $/$\varepsilon $ inhibitors PF-670462 and IC261 were also studied for comparison purposes.GSD0054 showed anti-proliferative activity against MDA-MB-453 and T-47D cells despite the fact that MDA-MB-453 cells do not possess active $\beta$-catenin. However, selective cell killing occurred in the more resistant, $\beta $-catenin active, T-47D cells. CK1$\varepsilon $ was confirmed as a cellular target, although other targets or alternative mechanisms of action could possibly explain the anti-proliferative activity in MDA-MB-453 cells.

Key words:  Synthetic lethality      Casein kinase inhibitors      Wnt/beta-catenin      Anti-beta-amino alcohols      Breast cancer     
Submitted:  24 December 2018      Revised:  03 January 2018      Accepted:  05 January 2018      Published:  20 April 2018     
*Corresponding Author(s):  José M. Padrón     E-mail:

Cite this article: 

Irene Lagunes, Elva Martín-Batista, Gastón Silveira-Dorta, Miguel X. Fernandes, José M. Padrón. Differential mechanism of action of the CK1$\varepsilon $ inhibitor GSD0054. Journal of Molecular and Clinical Medicine, 2018, 1(2): 77-84.

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Fig.1.  Chemical structures of ICK1$\varepsilon $ GSD0054 (1) and PF-4800567 (2); and ICK1$\delta $/$\varepsilon $ PF-670462 (3) and IC 261 (4).

Table 1  Anti-proliferative activity (GI$_{50})$ of ICK1$\varepsilon $ drugs in MDA-MB-453 ($\beta $-catenin negative) and T-47D ($\beta $-catenin positive) cell lines$^{\rm a}$.
MDA-MB-453 T-47D Rb
GSD0054 (1) 2.4 士 0.5 17 士 1.7 7.1
PF-4800567 (2) 58 士 11 79 士 6.4 1.4
PF-670462 (3) 42 士 6.5 11 士 2.0 3.8
IC 261 (4) 0.087 士 0.040 0.032 士 0.007 2.7
Table 2  Anti-proliferative activity (GI$_{50})$ of ICK1$\varepsilon $ in SW1573 and SW1573/P-gp cell lines$^{\rm a}$.
- Verapamil 十 Verapamil
SW1573 SW1573/P-gp Rfb SW1573 SW1573/P-gp Rf
GSD0054(1) 16 士 2 16 士 3 1 20 士 1 17 士 4 0.9
PF-4800567(2) 76 士 26 48 士 24 0.6 85 士 14 51 士 25 0.6
PF-670462 (3) 19 士 3 19 士 8 1 21 士 7 19 士 6 0.9
IC 261 (4) 0.25 士 0.03 0.79 士 0.10 3.2 0.35 士 0.03 0.86 士 0.42 2.5
Paclitaxel 0.0029 士 0.0009 4 士 0.5 1379 0.0030 士 0.0015 0.0036 士 0.0015 1.2
Fig.2.  Dose-response curves of SW1573 and SW1573/P-gp cells treated with ICK1$\varepsilon $ 1-4 for 48 h.

Fig.3.  Expression of $\beta $-catenin in drug-treated T-47D cells. Cells were exposed to compounds 1-4 for 24 h at the indicated doses. C (-): MDA-MB-453, negative control; C ($+$): T-47D, positive control. $\beta $-Actin signals allowed us to normalize the experiment.

Fig.4.  Cell cycle histograms of MDA-MB-453 and T-47D untreated cells (control) and cells treated (drug concentration in $\mu $M) for 24 h with compounds 1-4 at the indicated doses.

Fig.5.  ROS production increase in MDA-MB-453 and T-47D cells treated with drug (dose in $\mu $M) for 48 h with compounds 1-4 at the indicated concentrations. LD (low dose) of 1, 10, 10, 0.1 and HD (high dose) of 10, 100, 100, 1 $\mu $M for 1-4, respectively.

Table 3  Docking scores of drugs 1-4 against CK1$\varepsilon $.
Compound Score (kcal/mol)
GSD0054(1) -5.8
PF-4800567 (2) -10.8
PF-670462 (3) -6.5
IC 261 (4) -5.9
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