Supplementary Materials

This PDF file includes:

  • Fig. S1. Chemical structures of the novel small-molecule inhibitors of TNFR1 and a negative control compound.
  • Fig. S2. The seven hit compounds and zafirlukast bind the TNFR1 ECD as characterized by SPR measurements.
  • Fig. S3. Some hit compounds nonspecifically inhibit TNFR1-stimulated NF-κB activation.
  • Fig. S4. DS41, but not DS42 and zafirlukast, inhibits TRADD-induced NF-κB activation in HEK293 cells.
  • Fig. S5. Small-molecule inhibitors do not disrupt ligand-receptor interactions as characterized by SPR measurements.
  • Fig. S6. Hit compounds do not disrupt either the TNFR1 PLAD dimer or the LTα trimer.
  • Fig. S7. Hit compounds reduce FRET mediated by the TNFR1 mutant biosensors.
  • Fig. S8. Hit compounds compete with zafirlukast in binding to the TNFR1 PLAD and in inhibiting NF-κB activation.
  • Fig. S9. Hit compounds do not compete with the H398 antibody in modulating TNFR1 signaling.
  • Fig. S10. DSA114, an analog of DS41, shows improved potency and specificity by acting through the same noncompetitive inhibition mechanism.
  • Table S1. Functional characterization of DS41 and its analogs.

[Download PDF]