Ligand-based design identifies a potent NUPR1 inhibitor exerting anticancer activity via necroptosis
Patricia Santofimia-Castaño, … , José L. Neira, Juan Iovanna
Patricia Santofimia-Castaño, … , José L. Neira, Juan Iovanna
Published March 28, 2019
Citation Information: J Clin Invest. 2019;129(6):2500-2513. https://doi.org/10.1172/JCI127223.
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Research Article Gastroenterology Oncology

Ligand-based design identifies a potent NUPR1 inhibitor exerting anticancer activity via necroptosis

Abstract

Intrinsically disordered proteins (IDPs) are emerging as attractive drug targets by virtue of their prevalence in various diseases including cancer. Drug development targeting IDPs is challenging because IDPs have dynamic structure features and conventional drug design is not applicable. NUPR1 is an IDP that plays an important role in pancreatic cancer. We previously reported that trifluoperazine (TFP), an antipsychotic agent, was capable of binding to NUPR1 and inhibiting tumor growth. Unfortunately, TFP showed strong central nervous system side effects. In the present work, we undertook a multidisciplinary approach to optimize TFP based on the synergy of computer modeling, chemical synthesis, and a variety of biophysical, biochemical, and biological evaluations. A family of TFP-derived compounds was produced and the most active one, ZZW-115, showed a dose-dependent tumor regression with no neurological effects and an ability to induce cell death mainly by necroptosis. This study opens a new perspective for drug development against IDPs, demonstrating the possibility of successful ligand-based drug design for such challenging targets.

Authors

Patricia Santofimia-Castaño, Yi Xia, Wenjun Lan, Zhengwei Zhou, Can Huang, Ling Peng, Philippe Soubeyran, Adrián Velázquez-Campoy, Olga Abián, Bruno Rizzuti, José L. Neira, Juan Iovanna

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Figure 4

ZZW-115 induces cell death by necrosis and apoptosis in vitro.

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ZZW-115 induces cell death by necrosis and apoptosis in vitro. A panel o...
A panel of pancreatic cancer cell lines was incubated using 3 or 5 μM ZZW-115 for 24 hours and (A) LDH release and (B) caspase 3/7 activity were measured. Statistical significance is *P < 0.05 and ***P < 0.001 compared with untreated group (2-way ANOVA, Bonferroni’s post hoc test). (C) Flow cytometry analysis of annexin V/PI staining following 24 hours of treatment with 5 μM ZZW-115. A representative experiment of the dot plot profile of cells is shown (n = 3). MiaPaCa-2 cells were incubated from 0.5–5 μM ZZW-115 in the presence or absence of Z-VAD-FMK (20 μM) or/and Nec-1 (40 μM) for 24 hours, and (D) LDH release and (E) caspase 3/7 activity were measured. Statistical significance is *P < 0.05, **P < 0.01, and ***P < 0.001 compared with control group (2-way ANOVA, Bonferroni’s post hoc test). (F) Chemogram assays were done on pancreatic cancer cell lines with increasing concentrations of ZZW-115 in the presence or absence of Z-VAD-FMK (20 μM) or/and Nec-1 (40 μM) for 24 hours. AUC was calculated by integration. Statistical significance is **P < 0.01 and ***P < 0.001 compared with ZZW-115–treated cells (2-way ANOVA, Bonferroni’s post hoc test). MiaPaCa-2 cells were incubated in 5 μM ZZW-115 compound or 8 nM paclitaxel in the presence or absence of Z-VAD-FMK (20 μM), and (G) caspase 3/7 activity and (H) LDH release were measured. MiaPaCa-2 cells were incubated in 5 μM ZZW-115 or 8 nM paclitaxel in the presence or absence of Nec-1 (40 μM), and (I) caspase 3/7 activity and (J) LDH release were measured. Statistical significance is *P < 0.05, **P < 0.01, and ***P < 0.001 compared with ZZW-115–treated cells; #P < 0.05 and ##P < 0.01 compared with paclitaxel-treated cells (2-way ANOVA, Bonferroni’s post hoc test). Data represent mean ± SEM, n = 3.