Covalent Macrocyclic Proteasome Inhibitors Mitigate Resistance in Plasmodium falciparum

ACS Infect Dis. 2023 Oct 13;9(10):2036-2047. doi: 10.1021/acsinfecdis.3c00310. Epub 2023 Sep 15.

Abstract

The Plasmodium proteasome is a promising antimalarial drug target due to its essential role in all parasite lifecycle stages. Furthermore, proteasome inhibitors have synergistic effects when combined with current first-line artemisinin and related analogues. Linear peptides that covalently inhibit the proteasome are effective at killing parasites and have a low propensity for inducing resistance. However, these scaffolds generally suffer from poor pharmacokinetics and bioavailability. Here we describe the development of covalent, irreversible, macrocyclic inhibitors of the Plasmodium falciparum proteasome. We identified compounds with excellent potency and low cytotoxicity; however, the first generation suffered from poor microsomal stability. Further optimization of an existing macrocyclic scaffold resulted in an irreversible covalent inhibitor carrying a vinyl sulfone electrophile that retained high potency and low cytotoxicity and had acceptable metabolic stability. Importantly, unlike the parent reversible inhibitor that selected for multiple mutations in the proteasome, with one resulting in a 5,000-fold loss of potency, the irreversible analogue only showed a 5-fold loss in potency for any single point mutation. Furthermore, an epoxyketone analogue of the same scaffold retained potency against a panel of known proteasome mutants. These results confirm that macrocycles are optimal scaffolds to target the malarial proteasome and that the use of a covalent electrophile can greatly reduce the ability of the parasite to generate drug resistance mutations.

Keywords: Plasmodium falciparum;; antimicrobial resistance; cyclic peptides.

MeSH terms

  • Antimalarials* / chemical synthesis
  • Antimalarials* / chemistry
  • Antimalarials* / pharmacology
  • Drug Resistance*
  • Humans
  • Macrocyclic Compounds / chemistry
  • Macrocyclic Compounds / pharmacology
  • Malaria, Falciparum / drug therapy
  • Malaria, Falciparum / parasitology
  • Plasmodium falciparum* / drug effects
  • Plasmodium falciparum* / enzymology
  • Proteasome Endopeptidase Complex* / metabolism
  • Proteasome Inhibitors* / chemistry
  • Proteasome Inhibitors* / pharmacology
  • Sulfones / chemistry
  • Sulfones / pharmacology

Substances

  • Proteasome Inhibitors
  • Antimalarials
  • Proteasome Endopeptidase Complex
  • Macrocyclic Compounds
  • Sulfones