• Home
  • Search Results
  • Benzodiazepinone derivatives protect against endoplasmic reticulum stress-mediated cell death in human neuronal cell lines.

Benzodiazepinone derivatives protect against endoplasmic reticulum stress-mediated cell death in human neuronal cell lines.

ACS chemical neuroscience (2014-12-30)
Haixia Zou, Allison S Limpert, Jiwen Zou, Anna Dembo, Pooi-San Lee, Daniel Grant, Robert Ardecky, Anthony B Pinkerton, Gavin K Magnuson, Mark E Goldman, Juan Rong, Peter Teriete, Douglas J Sheffler, John C Reed, Nicholas D P Cosford
ABSTRACT

Endoplasmic reticulum (ER) stress causes neuronal dysfunction followed by cell death and is recognized as a feature of many neurodegenerative diseases. Using a phenotypic screen, we recently identified benzodiazepinone derivatives that reduce ER stress-mediated apoptosis in a rat neuronal progenitor cell line (CSM14.1). Herein we describe how structure-activity relationship (SAR) studies around these screening hits led to compounds that display robust cytoprotective activity against thapsigargin-induced ER stress in SH-SY5Y and H4 human neuronal cell lines. We demonstrate that the most potent of these derivatives, compound 4hh, inhibits the activation of p38 MAP kinase (p38) and c-Jun N-terminal kinase (JNK), protein kinases that are downstream signal effectors of the unfolded protein response (UPR). Compound 4hh specifically protects against thapsigargin-induced cell death and displays no protection against other insults known to induce cellular stress or activate p38. However, compound 4hh provides moderate inhibition of p38 activity stimulated by compounds that disrupt calcium homeostasis. Our data indicate that probe compound 4hh is a valuable small molecule tool that can be used to investigate the effects of ER stress on human neurons. This approach may provide the basis for the future development of therapeutics for the treatment of neurodegenerative diseases.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
RIPA Buffer
Sigma-Aldrich
DL-Dithiothreitol, ≥98% (HPLC), ≥99.0% (titration)
SAFC
BIS-TRIS
Sigma-Aldrich
DL-Dithiothreitol solution, BioUltra, for molecular biology, ~1 M in H2O
Supelco
DL-Dithiothreitol solution, 1 M in H2O
Sigma-Aldrich
Tunicamycin from Streptomyces sp.
Sigma-Aldrich
BIS-TRIS, ≥98.0% (titration)
Sigma-Aldrich
Thapsigargin, ≥98% (HPLC), solid film
Supelco
Paraquat dichloride hydrate, PESTANAL®, analytical standard
Sigma-Aldrich
6-Hydroxydopamine hydrochloride, ≥97% (titration), powder
Sigma-Aldrich
Z-Leu-Leu-Leu-al, ≥90% (HPLC)
Sigma-Aldrich
BIS-TRIS, BioXtra, ≥98.0% (titration)
Sigma-Aldrich
Bicinchoninic acid disodium salt hydrate, ≥98% (HPLC)
Sigma-Aldrich
BIS-TRIS, BioUltra, ≥99.0% (NT)
Sigma-Aldrich
BIS-TRIS, BioPerformance Certified, suitable for cell culture, suitable for insect cell culture, ≥98.0%
Supelco
Staurosporine, VETRANAL®, analytical standard
SAFC
BIS-TRIS