Potent and selective chemical probe of hypoxic signaling downstream of HIF-α hydroxylation via VHL inhibition

TY - JOUR

T1 - Potent and selective chemical probe of hypoxic signaling downstream of HIF-α hydroxylation via VHL inhibition

AU - Frost, Julianty

AU - Galdeano, Carles

AU - Soares, Pedro

AU - Gadd, Morgan S.

AU - Grzes, Katarzyna M.

AU - Ellis, Lucy

AU - Epemolu, Ola

AU - Shimamura, Satoko

AU - Bantscheff, Marcus

AU - Grandi, Paola

AU - Read, Kevin D.

AU - Cantrell, Doreen A.

AU - Rocha, Sonia

AU - Ciulli, Alessio

N1 - This work was supported by the European Research Council ERC-2012-StG-311460 DrugE3CRLs (Starting Grant to A.C.), the UK Biotechnology and Biological Sciences Research Council BBSRC BB/G023123/2 (David Phillips Fellowship to A.C.), the European Commission PIEF-GA-2012-328030 (Marie-Curie Intra-European Fellowship to C.G.), the Wellcome Trust (PhD Studentship 102398/Z/13/Z to J.F., and strategic awards 100476/Z/12/Z for biophysics and drug discovery and 094090/Z/10/Z for structural biology and X-ray crystallography to BCDD) and the Fundac¸a˜o para a Cieˆncia e Tecnologia FCT SFRH/BD/101598/2014 (PhD Studentship to P.S.). S.R. is funded by a Cancer Research UK Senior fellowship C99667/A12918 with support of a Wellcome Trust strategic award 097945/B/11/Z.

PY - 2016/11/4

Y1 - 2016/11/4

N2 - Chemical strategies to using small molecules to stimulate hypoxia inducible factors (HIFs) activity and trigger a hypoxic response under normoxic conditions, such as iron chelators and inhibitors of prolyl hydroxylase domain (PHD) enzymes, have broad-spectrum activities and off-target effects. Here we disclose VH298, a potent VHL inhibitor that stabilizes HIF-α and elicits a hypoxic response via a different mechanism, that is the blockade of the VHL:HIF-α protein-protein interaction downstream of HIF-α hydroxylation by PHD enzymes. We show that VH298 engages with high affinity and specificity with VHL as its only major cellular target, leading to selective on-target accumulation of hydroxylated HIF-α in a concentration- and time-dependent fashion in different cell lines, with subsequent upregulation of HIF-target genes at both mRNA and protein levels. VH298 represents a high-quality chemical probe of the HIF signaling cascade and an attractive starting point to the development of potential new therapeutics targeting hypoxia signaling.

AB - Chemical strategies to using small molecules to stimulate hypoxia inducible factors (HIFs) activity and trigger a hypoxic response under normoxic conditions, such as iron chelators and inhibitors of prolyl hydroxylase domain (PHD) enzymes, have broad-spectrum activities and off-target effects. Here we disclose VH298, a potent VHL inhibitor that stabilizes HIF-α and elicits a hypoxic response via a different mechanism, that is the blockade of the VHL:HIF-α protein-protein interaction downstream of HIF-α hydroxylation by PHD enzymes. We show that VH298 engages with high affinity and specificity with VHL as its only major cellular target, leading to selective on-target accumulation of hydroxylated HIF-α in a concentration- and time-dependent fashion in different cell lines, with subsequent upregulation of HIF-target genes at both mRNA and protein levels. VH298 represents a high-quality chemical probe of the HIF signaling cascade and an attractive starting point to the development of potential new therapeutics targeting hypoxia signaling.

U2 - 10.1038/ncomms13312

DO - 10.1038/ncomms13312

M3 - Article

C2 - 27811928

SN - 2041-1723

VL - 7

SP - 1

EP - 12

JO - Nature Communications

JF - Nature Communications

M1 - 13312

ER -