Abstract

Introduction Pancreatic cancer continues to be among the most lethal malignancies. The distinct tumor microenvironment (TME) not only facilitates tumor cell proliferation, invasion, and metastasis but also establishes a formidable barrier to the diffusion of chemotherapeutic agents and immunotherapies. Consequently, it is imperative to identify novel compounds that target the tumor microenvironment (TME). Objectives This study aims to identify a small molecule as a therapeutic agent for pancreatic cancer through dual mechanisms, specifically DDR1 inhibition and methuosis induction. Methods We characterized DDR1 as a primary target of N-(4-Methyl-3-(4-(5-(4-methylpiperazin-1-yl)pyridin-3-yl)-1H-pyrazol-1-yl)phenyl)-6-(trifluoromethyl)picolinamide (IHMT-140) using ADP-Glo assays, KINOMEscan, Western blotting, and molecular docking. Colony formation, invasion, wound-healing migration, and immunohistochemical staining assays demonstrated that IHMT-140 suppresses epithelial-mesenchymal transition (EMT) by regulating key EMT markers. Cell viability assays, including Cell Titer-Glo and live-cell imaging, transmission electron microscopy (TEM), and immunofluorescence staining, revealed that IHMT-140 triggers methuosis in pancreatic cancer cells. RNA-seq was also employed to confirm the underlying mechanisms. Finally, Cell Titer-Glo, Western blot, and flow cytometry were applied to demonstrate the combined antitumor effects of IHMT-140 and gemcitabine. Gemcitabine uptake and tumor accumulation were quantified by high-resolution mass spectrometry. The in vivo efficacy was tested in a xenograft model, with tumor sections analyzed by H&E and pan-collagen staining. Results We discovered N-(4-Methyl-3-(4-(5-(4-methylpiperazin-1-yl)pyridin-3-yl)-1H-pyrazol-1-yl)phenyl)-6-(trifluoromethyl)picolinamide (IHMT-140) as a novel dual-function agent that acts as a DDR1 inhibitor and a methuosis inducer. It simultaneously remodels the tumor stromal ECM and activates excessive macropinocyt