An Innovative High-Throughput Screening Approach for Discovery of Small Molecules That Inhibit TNF Receptors

SLAS Discovery
Chih Hung LoJonathan N Sachs

Abstract

Tumor necrosis factor receptor 1 (TNFR1) is a transmembrane receptor that binds tumor necrosis factor or lymphotoxin-alpha and plays a critical role in regulating the inflammatory response. Upregulation of these ligands is associated with inflammatory and autoimmune diseases. Current treatments reduce symptoms by sequestering free ligands, but this can cause adverse side effects by unintentionally inhibiting ligand binding to off-target receptors. Hence, there is a need for new small molecules that specifically target the receptors, rather than the ligands. Here, we developed a TNFR1 FRET biosensor expressed in living cells to screen compounds from the NIH Clinical Collection. We used an innovative high-throughput fluorescence lifetime screening platform that has exquisite spatial and temporal resolution to identify two small-molecule compounds, zafirlukast and triclabendazole, that inhibit the TNFR1-induced IκBα degradation and NF-κB activation. Biochemical and computational docking methods were used to show that zafirlukast disrupts the interactions between TNFR1 pre-ligand assembly domain (PLAD), whereas triclabendazole acts allosterically. Importantly, neither compound inhibits ligand binding, proving for the first time tha...Continue Reading

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Citations

Oct 28, 2019·Alzheimer's & Dementia : the Journal of the Alzheimer's Association·Chih Hung LoJonathan N Sachs
Jan 22, 2020·Protein Science : a Publication of the Protein Society·Chih Hung LoJonathan N Sachs
Aug 1, 2019·Science Signaling·Penny E MortonMaddy Parsons
Aug 1, 2019·Science Signaling·Chih Hung LoJonathan N Sachs
Jun 13, 2020·Frontiers in Cell and Developmental Biology·Roman FischerKlaus Pfizenmaier
May 13, 2018·International Journal of Molecular Sciences·Sophie SteelandRoosmarijn E Vandenbroucke
Oct 26, 2018·Biosensors·Tory M SchaafGregory D Gillispie
Jun 3, 2021·International Journal of Molecular Sciences·Andy RuizLeslie Chavez-Galan
Sep 18, 2020·Biochemistry·Nagamani VunnamBenjamin J Hackel

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Methods Mentioned

BETA
fluorescence
biosensor
FRET
flow
fluorescence microscopy
flow cytometry
Assay
electrophoresis
immunoprecipitation
size-exclusion

Software Mentioned

MATLAB
WSA
Dock
MOE Dock
FlowJo
MOE
Fluorescence Innovations
GBVI
Image Studio
Evotec

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