XMD-17-51 Inhibits DCLK1 Kinase and Prevents Lung Cancer Progression.

Wei-Qiang Yang,Xue-Lin Zhang,Guang Chen,Alexander Kiselyov,Shuai-Jun Xu,Liu-Lian Zhu,Wei-Jun Zhao,Xiao-Fei Ding,Yong Liang

doi:10.3389/fphar.2021.603453

Wei-Qiang Yang, Xue-Lin Zhang + Show 7 more

Open Access

https://doi.org/10.3389/fphar.2021.603453

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Abstract

Doublecortin-like kinase 1 (DCLK1) is a cancer stem cell marker that is highly expressed in various types of human cancer, and a protein kinase target for cancer therapy that is attracting increasing interest. However, no drug candidates targeting DCLK1 kinase have been developed in clinical trials to date. XMD-17-51 was found herein to possess DCLK1 kinase inhibitory activities by cell-free enzymatic assay. In non-small cell lung carcinoma (NSCLC) cells, XMD-17-51 inhibited DCLK1 and cell proliferation, while DCLK1 overexpression impaired the anti-proliferative activity of XMD-17-51 in A549 cell lines. Consequently, XMD-17-51 decreased Snail-1 and zinc-finger-enhancer binding protein 1 protein levels, but increased those of E-cadherin, indicating that XMD-17-51 reduces epithelial-mesenchymal transition (EMT). Furthermore, sphere formation efficiency was significantly decreased upon XMD-17-51 treatment, and XMD-17-51 reduced the expression of stemness markers such as β-catenin, and pluripotency factors such as SOX2, NANOG and OCT4. However, the percentage of ALDH+ cells was increased significantly following treatment with XMD-17-51 in A549 cells, possibly due to EMT inhibition. In combination, the present data indicated that XMD-17-51 inhibited DCLK1 kinase activity in a cell-free assay with an IC50 of 14.64 nM, and decreased DCLK1 protein levels, cell proliferation, EMT and stemness in NSCLC cell lines. XMD-17-51 has the potential to be a candidate drug for lung cancer therapy.

Highlights

XMD-17-51 trifluoroacetate, a pyrimido-diazepinone compound, was originally discovered as a derivative of HTH-01-015, which is a selective inhibitor of NUAK1 (Banerjee et al, 2014; Gray et al, 2016)
To confirm the inhibition of Doublecortin-like kinase 1 (DCLK1) kinase in vitro, A549 cells were treated with XMD-17-51 at different concentrations for 24 h
DCLK1 is highly expressed in several malignancies and is a marker of tumor stem cells that plays a critical role in the self-renewal capacity of cells derived from these tumors (Panneerselvam et al, 2020)

Summary

Introduction

XMD-17-51 trifluoroacetate, a pyrimido-diazepinone compound, was originally discovered as a derivative of HTH-01-015, which is a selective inhibitor of NUAK1 (Banerjee et al, 2014; Gray et al, 2016). XMD-17-51 has been shown to inhibit several members of the AMPK family (MARK1, MARK3, BRSK1, and AMPK) and kinases associated with growth and proliferation (Banerjee et al, 2014). DCLK1, a member of the protein kinase superfamily and the dipcortin family was initially found to be XMD-17-51 Inhibits DCLK1 Kinase associated with microtubules involved in neurogenesis and neuronal migration (Lin et al, 2000). DCLK1 was found to be highly expressed in several malignancies and to be involved in the regulation of tumorigenesis, tumor stemness and epithelialmesenchymal transition (EMT) in cancer, including pancreatic, colorectal and liver cancer (Chandrakesan et al, 2014; Weygant et al, 2015; Nguyen et al, 2016; Westphalen et al, 2016; Weygant et al, 2016; Chandrakesan et al, 2017; Liu et al, 2018). The expression and biological function of DCLK1 in non-small cell lung carcinoma (NSCLC) remain unclear. DCLK1 kinase inhibitors have neither been evaluated in clinical trials nor been approved for clinical therapy

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