Monomerization of Homodimeric Trefoil Factor 3 (TFF3) by an Aminonitrile Compound Inhibits TFF3-Dependent Cancer Cell Survival

Abstract

Trefoil factor 3 (TFF3) is a secreted protein with an established oncogenic function and a highly significant association with clinical progression of various human malignancies. Herein, a novel small molecule that specifically targets TFF3 homodimeric functions was identified. Utilizing the concept of reversible covalent interaction, 2-amino-4-(4-(6-fluoro-5-methylpyridin-3-yl)­phenyl)-5-oxo-4H,5H-pyrano­[3,2-c]­chromene-3-carbonitrile (AMPC) was identified as a molecule that interacted with TFF3. AMPC monomerized the cellular and secreted TFF3 homodimer at the cysteine (Cys)­57-Cys57 residue with subsequent more rapid degradation of the generated TFF3 monomers. Hence, AMPC treatment also resulted in cellular depletion of TFF3 with consequent decreased cell viability in various human carcinoma-derived TFF3 expressing cell lines, including estrogen receptor positive (ER+) mammary carcinoma (MC). AMPC treatment of TFF3 expressing ER+ MC cells significantly suppressed total cell number in a dose-dependent manner. Consistently, exposure of TFF3 expressing ER+ MC cells to AMPC decreased soft agar colony formation, foci formation, and growth in suspension culture and inhibited growth of preformed colonies in 3D Matrigel. AMPC increased apoptosis in TFF3 expressing ER+ MC cells associated with decreased activity of EGFR, p38, STAT3, AKT, and ERK, decreased protein levels of CCND1, CCNE1, BCL2, and BCL-XL, and increased protein levels of TP53, CDKN1A, CASP7, and CASP9. siRNA-mediated depletion of TFF3 expression in ER+ MC cells efficiently abrogated AMPC-stimulated loss of cell viability and CASPASE 3/7 activities. Furthermore, in mice bearing ER+ MC cell-generated xenografts, AMPC treatment significantly impeded xenograft growth. Hence, AMPC exemplifies a novel mechanism by which small molecule drugs may inhibit a dimeric oncogenic protein and provides a strategy to impede TFF3-dependent cancer progression.

Vertical pathway inhibition of receptor tyrosine kinases and BAD with synergistic efficacy in triple negative breast cancer

“…Molecular interactions were analyzed by SPR using a BIAcore-2000 system (BIAcore AB, Uppsala, Sweden). Recombinant human BAD (Novoprotein, Suzhou, China) was immobilized on a sensor chip and analyzed as per the manufacturer’s protocol and as previously described 73 . Combination index (CI) analysis was performed using the Chou-Talalay 74 and SynergyFinder 75 CI analysis method.…”

Discovery of Pyrimidine- and Coumarin-Linked Hybrid Molecules as Inducers of JNK Phosphorylation through ROS Generation in Breast Cancer Cells

“…The novel PCL compounds were initially subjected to cell viability studies against human breast cancer cells (MCF-7) using an MTT assay [18,19] for 24 h [20,21]. PC-12 exhibited the most potent loss of cell viability against BT-474 cells with an IC 50 of 32 µM.…”

Electrochemical Synthesis of Versatile Pyrimidine and Oxadiazoles Tethered Triazoles as Inhibitors of VEGFR-2 in Human Breast Cancer Cells

“…Heterocycles have been widely studied and integrated into medicinal chemistry due to their diverse biological activities, including targeting specific cellular pathways, DNA binding, alkylation, and apoptosis induction [4,5]. Presently, extensive research is focusing on various natural and synthetic heterocyclic compounds, such as pyrimidines [6][7][8], coumarins [9,10], pyrazoles [11][12][13], triazoles [14][15][16], oxadiazoles [17][18][19], and piperazines [20,21], among others, which have displayed promising biological properties.…”

“…Triazoles, known for their structural diversity and kinase interactions, are extensively studied for medicinal chemistry. Benzimidazole-substituted triazoles, e.g., compound (10), are potent inhibitors of VEGFR-2. Zebrafish studies demonstrate the more substantial anti-angiogenic potential of indole-2-one substituted 1,2,3-triazole (11) than that of sunitinib.…”