Expanding the Structural Diversity at the Phenylene Core of Ligands for the von Hippel-Lindau (VHL) E3 Ubiquitin Ligase: Development of Highly Potent Hypoxia-Inducible Factor-1alpha (HIF-1alpha) Stabilizers

01 August 2023, Version 2
This content is a preprint and has not undergone peer review at the time of posting.


Hypoxia-inducible factor-1α (HIF-1α) constitutes the principal mediator of cellular adaptation to hypoxia in humans. HIF-1α protein level and activity is tightly regulated by the ubiquitin E3 ligase von Hippel-Lindau (VHL). Here, we performed a structure-guided and bioactivity-driven design of new VHL inhibitors. Our iterative and combinatorial strategy focused on chemical variability at the phenylene unit and encompassed further points of diversity. The exploitation of tailored phenylene fragments and the stereoselective installation of the benzylic methyl group provided potent VHL ligands. Three high-resolution structures of VHL-ligand complexes were determined and bioactive conformations of these ligands were explored. The most potent inhibitor (30) exhibited dissociation constants lower than 40 nM, independently determined by fluorescence polarization and surface plasmon resonance and an enhanced cellular potency, as evidenced by its superior ability to induce HIF-1α transcriptional activity. Our work is anticipated to inspire future efforts towards HIF-1α stabilizers and new ligands for proteolysis-targeting chimeras.


von Hippel-Lindau
E3 ligase
Hypoxia-Inducible Factor
protein-protein interaction
drug design
X-ray crystallography
structure-based drug design

Supplementary materials

Supporting Information
Supporting Figures S1-S9. Supporting Tables S1-S2: mean IC50 values obtained from the FP assay; values kon and koff for selected VHL ligands determined by SPR. Supporting Schemes S1-S2: Synthesis of building blocks 41α-ε. 1H and 13C NMR Assignments. 1H and 13C NMR Spectra. LC-MS Traces of Final Compounds. Crystallographic Data for Co-crystal Structures of VCB in Complex with 30, 33, and 37.


Comments are not moderated before they are posted, but they can be removed by the site moderators if they are found to be in contravention of our Commenting Policy [opens in a new tab] - please read this policy before you post. Comments should be used for scholarly discussion of the content in question. You can find more information about how to use the commenting feature here [opens in a new tab] .
This site is protected by reCAPTCHA and the Google Privacy Policy [opens in a new tab] and Terms of Service [opens in a new tab] apply.