Addressing the Intracellular Vestibule of the Plasmodial Lactate Transporter PfFNT by p-Substituted Inhibitors Amplifies In Vitro Activity

J Med Chem. 2024 Oct 24;67(20):18368-18383. doi: 10.1021/acs.jmedchem.4c01674. Epub 2024 Oct 3.

Abstract

Inhibition of the lactate transporter PfFNT is a valid novel mode of action against malaria parasites. Current pyridine-substituted pentafluoro-3-hydroxy-pent-2-en-1-ones act as substrate analogs with submicromolar EC50 in vitro, and >99.7% activity in mice. The recently solved structure of a PfFNT-inhibitor complex visualized the binding mode. Here, we extended the inhibitor layout by series of amine- and anilide-linked pyridine p-substituents to generate additional interactions in the cytoplasmic vestibule. Virtual docking indicated hydrogen bonding to Tyr31 and Ser102. Fluorescence cross-correlation spectroscopy yielded respectively enhanced target affinity. Strikingly, the in vitro activity increased by 1 order of magnitude to 14.8 nM at negligible cytotoxicity. While p-amine substitutions were rapidly metabolized, the more stable p-acetanilide cleared 99.7% of parasites at 4 × 50 mg kg-1 in a mouse malaria model. Future stabilization of the p-substitution against metabolism may translate the gain in in vitro potency to the in vivo situation.

MeSH terms

  • Alkenes / chemistry
  • Alkenes / pharmacology
  • Animals
  • Antimalarials* / chemical synthesis
  • Antimalarials* / chemistry
  • Antimalarials* / pharmacology
  • Humans
  • Malaria / drug therapy
  • Mice
  • Molecular Docking Simulation
  • Monocarboxylic Acid Transporters / antagonists & inhibitors
  • Monocarboxylic Acid Transporters / metabolism
  • Plasmodium falciparum* / drug effects
  • Plasmodium falciparum* / metabolism
  • Protozoan Proteins / antagonists & inhibitors
  • Protozoan Proteins / chemistry
  • Protozoan Proteins / metabolism
  • Pyridines / chemical synthesis
  • Pyridines / chemistry
  • Pyridines / pharmacology
  • Structure-Activity Relationship

Substances

  • Antimalarials
  • Monocarboxylic Acid Transporters
  • Protozoan Proteins
  • Pyridines
  • Alkenes