Designing selectives for one of six ion channel isoforms

Achieving in vivo PoC with the first ‘shot’

40%

hits active
in vitro

8 fold

isoform/conformation
selectivity

3

compounds active
in vivo

2

completely new classes
of compounds

*Project methodology

01/ Background

  • The target is in the family of highly similar ion channel isoforms.
  • The goal is to develop selective inhibitors specifically to the open channel conformation.
  • The exact location of the most favorable binding pocket for achieving selectivity is unknown.
  • In vivo PoC was essential to confirm therapeutic efficacy.

02/ Methodology

  • 3 tentative binding sites were identified for each isoform by
proprietary Receptor.AI pocket detection workflow:
    in the outer channel pore, inside the channel cavity,
    and between the functionally important transmembrane helices.
  • Selectivity assessment was performed based on differential
    pocket pharmacophore and generative AI binding pose prediction.
  • Stock chemical space of 8M compounds was used as well as a custom focused diversity database of 50K compounds.
  • 291 hit candidates were selected for experimental validation.

03/ Metrics

  • Fold increase of effect on target isoform compared to off-targets.
  • UFD effect: the preference of the compound to block the active channel state relative to the resting state.
  • Peak blocking of the target channel at 120 μM of the compound relative to the vehicle.

04/ Results

  • 40% of hit candidates validated active in vitro.
  • 5 highly selective hits found, with the top hit compound exhibiting
8.6x selectivity.
  • 3 compounds demonstrated substantial in vivo activity
    (murine model) and were proven to be non-toxic.
*Similarity matrix of full-length isoforms
*Results of in vitro experimental assessments for top 5 compounds