In vitro evaluation of dihydropyridine-3-carbonitriles as potential cytotoxic agents through PIM-1 protein kinase inhibition

Khalil Abnous, Hesam Manavi, Soghra Mehri, Mona Alibolandi, Hossein Kamali, Morteza Ghandadi, Farzin Hadizadeh


PIM-1 protein kinase inhibitor belongs to a novel class of serine/threonine kinases. As PIM-1 is overexpressed in cancer cells and possesses oncogenic functions, its inhibition provides a new option in cancer therapy. In this study, in vitro inhibitory effects of seven analogues of 1, 2-dihydropyridine-3-carbonitrile derivatives Ia-c, IIa-d on the activity of recombinant PIM-1 were evaluated using dimethylthiazol diphenyltetrazolium bromide (MTT) assay. The PIM-1 protein kinase inhibitory potencies and the cytotoxicity effects of tested compounds were respectively as follows: Ic > IIa > Ia > IIb > Ib > IId > IIc and IIb > IIa > Ia > IIc > Ic > Ib > IId, respectively. The compound Ic with methylthio imidazole substituent at C-3 position and benzodioxole substituent at C-6 position of 2-imino-1, 2-dihydropyridine-3- carbonitrile structure showed the strongest PIM-1 inhibitory effect (IC50 = 111.01 nM), while the compound IIc with methythio imidazole substituent at C-3 position and benzodioxole substituent at C-6 position of             2-oxo-1, 2-dihydropyridine-3- carbonitrile structure exhibited the least inhibition activity (IC50 = 433.71 nM). The docking results showed that all tested compounds localized appropriately in the middle of binding cavity after docking procedure, demonstrating suitable interactions between ligands and protein. This study demonstrated that the PIM-1 inhibitory potencies of newly synthesized compounds were in submicromolar concentrations (IC50 < 150 nM) while they exhibited low cytotoxicity on HT-29 cell line (IC50> 130 µM). Altogether, our data indicated that compounds Ic, IIa, Ia could be considered as new potent non-toxic PIM-1 inhibitors which could be used in combination with routine anti-proliferative drugs.


PIM-1; Protein kinase inhibition; Dihydropyridine-3- carbonitrile; Anticancer; Cytotoxicity

Full Text:



Qian KC, Wang L, Hickey ER, Studts J, Barringer K, Peng C, et al. Structural basis of constitutive activity and a unique nucleotide binding mode of human Pim-1 kinase. J Biol Chem. 2005;280(7):6130-6137.

Larid PW, Van der Lugt NM, Clarke A, Domen J, Linders K, McWhir J, et al. In vivo analysis of Pim-1 deficiency. Nucleic Acids Res. 1993;21(20):4750-4755.

Amaravadi R, Thompson CB. The survival kinases Akt and Pim as potential pharmacological targets. J clin Invest. 2005;115(10):2618-2624.

Möröy T, Grzeschiczek A, Petzold S, Hartmann K-U. Expression of a Pim-1 transgene accelerates lymphoproliferation and inhibits apoptosis in lpr/lpr mice. Proc Natl Acad Sci U S A. 1993;90(22):10734-10738.

Jung YJ, Chae HC, Seoh JY, Ryu KH, Park HK, Kim YJ, et al. Pim‐1 induced polyploidy but did not affect megakaryocytic differentiation of K562 cells and CD34+ cells from cord blood. Eur J Haematol. 2007;78(2):131-138.

Merkel AL, Meggers E, Ocker M. PIM1 kinase as a target for cancer therapy. Expert Opin Investig Drugs. 2012;21(4):425-436.

Shah N, Pang B, Yeoh K-G, Thorn S, Chen CS, Lilly MB, et al. Potential roles for the PIM1 kinase in human cancer–a molecular and therapeutic appraisal. Eur J Cancer. 2008;44(15):2144-2151.

Cibull T, Jones T, Li L, Eble J, Baldridge LA, Malott S, et al. Overexpression of Pim-1 during progression of prostatic adenocarcinoma. J Clin Pathol. 2006;59(3):285-288.

Narlik‐Grassow M, Blanco‐Aparicio C, Carnero A. The PIM family of serine/threonine kinases in cancer. Med Res Rev. 2014;34(1):136-159.

Nawijn MC, Alendar A, Berns A. For better or for worse: the role of Pim oncogenes in tumorigenesis. Nat Rev Cancer. 2011;11(1):23-34.

Mochizuki T, Kitanaka C, Noguchi K, Muramatsu T, Asai A, Kuchino Y. Physical and functional interactions between pim-1 kinase and cdc25a phosphatase implications for the pim-1-mediated activation of the c-myc signaling pathway. J Biol Chem. 1999;274(26):18659-18666.

Leverson JD, Koskinen PJ, Orrico FC, Rainio E-M, Jalkanen KJ, Dash AB, et al. Pim-1 kinase and p100 cooperate to enhance c-Myb activity. Mol Cell. 1998;2(4):417-425.

Rainio E-M, Sandholm J, Koskinen PJ. Cutting edge: Transcriptional activity of NFATc1 is enhanced by the Pim-1 kinase. J Immunol. 2002;168(4):1524-1527.

Aho TL, Sandholm J, Peltola KJ, Ito Y, Koskinen PJ. Pim-1 kinase phosphorylates RUNX family transcription factors and enhances their activity. BMC Cell Biol. 2006;7(1):21.

Aho TL, Sandholm J, Peltola KJ, Mankonen HP, Lilly M, Koskinen PJ. Pim‐1 kinase promotes inactivation of the pro‐apoptotic Bad protein by phosphorylating it on the Ser112 gatekeeper site. FEBS Lett. 2004;571(1-3):43-49.

Isaac M, Siu A, Jongstra J. The oncogenic PIM kinase family regulates drug resistance through multiple mechanisms. Drug Resist Updat. 2011;14(4):203-211.

Hu XF, Li J, Vandervalk S, Wang Z, Magnuson NS, Xing PX. PIM-1–specific mAb suppresses human and mouse tumor growth by decreasing PIM-1 levels, reducing Akt phosphorylation, and activating apoptosis. J Clin Invest. 2009;119(2):362-375.

Hsu J-L, Leong P-K, Ho Y-F, Hsu L-C, Lu P-H, Chen C-S, et al. Pim-1 knockdown potentiates paclitaxel-induced apoptosis in human hormone-refractory prostate cancers through inhibition of NHEJ DNA repair. Cancer Lett. 2012;319(2):214-222.

Jacobs MD, Black J, Futer O, Swenson L, Hare B, Fleming M, et al. Pim-1 ligand-bound structures reveal the mechanism of serine/threonine kinase inhibition by LY294002. J Biol Chem. 2005;280(14):13728-13734.

Debreczeni JÉ, Bullock AN, Atilla GE, Williams DS, Bregman H, Knapp S, et al. Ruthenium Half‐Sandwich Complexes Bound to Protein Kinase Pim‐1. Angew Chem Int Ed Engl. 2006;45(10):1580-1585.

Abadi AH, Ibrahim TM, Abouzid KM, Lehmann J, Tinsley HN, Gary BD, et al. Design, synthesis and biological evaluation of novel pyridine derivatives as anticancer agents and phosphodiesterase 3 inhibitors. Bioorg Med Chem. 2009;17(16):5974-5982.

Davari AS, Abnous K, Mehri S, Ghandadi M, Hadizadeh F. Synthesis and biological evaluation of novel pyridine derivatives as potential anticancer agents and phosphodiesterase-3 inhibitors. Bioorg Chem. 2014;57:83-89.

Badieyan ZS, Moallem SA, Mehri S, Shahsavand S, Hadizadeh F. Virtual screening for finding novel COX-2 inhibitors as antitumor agents. Open Med Chem J. 2012;6:15-19.

Guo S, Mao X, Chen J, Huang B, Jin C, Xu Z, et al. Overexpression of Pim-1 in bladder cancer. J Exp Clin Cancer Res. 2010;29:161.

Moreau P, G Dezhenkova L, Anizon F, Nauton L, Thery V, Liang S, et al. New potent and selective inhibitor of Pim-1/3 protein kinases sensitizes human colon carcinoma cells to doxorubicin. Anti-Cancer Agent Med Chem. 2014;14(9):1228-1236.


  • There are currently no refbacks.