Macrocyclic Modalities Combining Peptide Epitopes and Natural Product Fragments

J Am Chem Soc. 2020 Mar 11;142(10):4904-4915. doi: 10.1021/jacs.0c00269. Epub 2020 Mar 2.


"Hot loop" protein segments have variable structure and conformation and contribute crucially to protein-protein interactions. We describe a new hot loop mimicking modality, termed PepNats, in which natural product (NP)-inspired structures are incorporated as conformation-determining and -restricting structural elements into macrocyclic hot loop-derived peptides. Macrocyclic PepNats representing hot loops of inducible nitric oxide synthase (iNOS) and human agouti-related protein (AGRP) were synthesized on solid support employing macrocyclization by imine formation and subsequent stereoselective 1,3-dipolar cycloaddition as key steps. PepNats derived from the iNOS DINNN hot loop and the AGRP RFF hot spot sequence yielded novel and potent ligands of the SPRY domain-containing SOCS box protein 2 (SPSB2) that binds to iNOS, and selective ligands for AGRP-binding melanocortin (MC) receptors. NP-inspired fragment absolute configuration determines the conformation of the peptide part responsible for binding. These results demonstrate that combination of NP-inspired scaffolds with peptidic epitopes enables identification of novel hot loop mimics with conformationally constrained and biologically relevant structure.

MeSH terms

  • Agouti-Related Protein / chemistry
  • Agouti-Related Protein / metabolism
  • Epitopes
  • Humans
  • Nitric Oxide Synthase Type II / chemistry
  • Nitric Oxide Synthase Type II / metabolism
  • Peptides, Cyclic / chemical synthesis
  • Peptides, Cyclic / chemistry
  • Peptides, Cyclic / metabolism*
  • Protein Binding
  • Protein Conformation
  • Receptors, Melanocortin / metabolism*
  • Stereoisomerism
  • Suppressor of Cytokine Signaling Proteins / metabolism*


  • AGRP protein, human
  • Agouti-Related Protein
  • Epitopes
  • Peptides, Cyclic
  • Receptors, Melanocortin
  • SPSB2 protein, human
  • Suppressor of Cytokine Signaling Proteins
  • NOS2 protein, human
  • Nitric Oxide Synthase Type II