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 June 3, 2019; First 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 5

ZZW-115 treatment induces a decrease in ATP production and induces a ROS overproduction.

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ZZW-115 treatment induces a decrease in ATP production and induces a ROS...
(A) ATP content measured in a panel of pancreatic cancer cell lines incubated at 3 or 5 μM ZZW-115 for 24 hours. (B) OXPHOS metabolism, reflected by oxygen consumption rate (OCR) levels for basal respiration (B. resp.), maximal respiration (M. resp.), spare capacity (Spare cap.), and ATP production (ATP prod.) and (C) anaerobic glycolytic metabolism reflected by extracellular acidification rate (ECAR) levels for glycolysis (Glyco.), glycolytic capacity (G. cap.), and glycolysis reserve (G. res.) were measured in MiaPaCa-2 cells treated with 3 μM ZZW-115 compound for 4, 8, or 24 hours. (D) ATP production by OXPHOS and anaerobic glycolysis were determined in MiaPaCa-2 cells treated with 3 μM ZZW-115 for 4, 8, or 24 hours. Statistical significance is *P < 0.05, **P < 0.01 and ***P < 0.001 for OXPHOS or glycolysis compared with control cells; #P < 0.05 and ###P < 0.001 for total ATP compared with control cells (2-way ANOVA, Bonferroni’s post hoc test). (E) ROS production was detected using MitoSOX Red by flow cytometry analysis on a panel of pancreatic cancer cell lines incubated with ZZW-115 at 3 μM. Statistical significance: *P < 0.05, **P < 0.01, and ***P < 0.001 compared with control cells (Student’s 2-tailed unpaired t test). (F) MiaPaCa-2 cells were incubated at a ZZW-115 concentration range of 0.5–5 μM in the presence or absence of Z-VAD-FMK (20 μM) or/and Nec-1 (40 μM) for 24 hours, and ATP content was measured. (G and H) MiaPaCa-2 cells were incubated with 5 μM ZZW-115 or 8 nM paclitaxel in the presence or absence of Z-VAD-FMK (20 μM) or Nec-1 (40 μM), and ATP content was measured. Statistical significance: *P < 0.05, **P < 0.01, and ***P < 0.001 compared with ZZW-115–treated cells; #P < 0.05 and ###P < 0.001 compared with paclitaxel-treated cells (2-way ANOVA, Bonferroni’s post hoc test). Data represent mean ± SEM, n = 3.