Lipid Trolling to Optimize A3 Adenosine Receptor-Positive Allosteric Modulators (PAMs)

J Med Chem. 2024 Jul 25;67(14):12221-12247. doi: 10.1021/acs.jmedchem.4c00944. Epub 2024 Jul 3.

Abstract

A3 adenosine receptor (A3AR) positive allosteric modulators (PAMs) (2,4-disubstituted-1H-imidazo[4,5-c]quinolin-4-amines) allosterically increase the Emax of A3AR agonists, but not potency, due to concurrent orthosteric antagonism. Following mutagenesis/homology modeling of the proposed lipid-exposed allosteric binding site on the cytosolic side, we functionalized the scaffold, including heteroatom substitutions and exocyclic phenylamine extensions, to increase allosteric binding. Strategically appended linear alkyl-alkynyl chains with terminal amino/guanidino groups improved allosteric effects at both human and mouse A3ARs. The chain length, functionality, and attachment position were varied to modulate A3AR PAM activity. For example, 26 (MRS8247, p-alkyne-linked 8 methylenes) and homologues increased agonist Cl-IB-MECA's Emax and potency ([35S]GTPγS binding). The putative mechanism involves a flexible, terminally cationic chain penetrating the lipid environment for stable electrostatic anchoring to cytosolic phospholipid head groups, suggesting "lipid trolling", supported by molecular dynamic simulation of the active-state model. Thus, we have improved A3AR PAM activity through rational design based on an extrahelical, lipidic binding site.

MeSH terms

  • Adenosine A3 Receptor Agonists* / chemistry
  • Adenosine A3 Receptor Agonists* / pharmacology
  • Allosteric Regulation / drug effects
  • Allosteric Site
  • Animals
  • CHO Cells
  • Cricetulus
  • Humans
  • Lipids / chemistry
  • Mice
  • Quinolines / chemical synthesis
  • Quinolines / chemistry
  • Quinolines / pharmacology
  • Receptor, Adenosine A3* / chemistry
  • Receptor, Adenosine A3* / metabolism
  • Structure-Activity Relationship

Substances

  • Receptor, Adenosine A3
  • Adenosine A3 Receptor Agonists
  • Lipids
  • Quinolines