Evidence for the Capability of Roxadustat (FG-4592), an Oral HIF Prolyl-Hydroxylase Inhibitor, to Perturb Membrane Ionic Currents: An Unidentified yet Important Action.

Wei-Ting Chang,Sheng-Nan Wu,Zi-Han Gao,Yi-Ching Lo

doi:10.3390/ijms20236027

Abstract

Roxadustat (FG-4592), an analog of 2-oxoglutarate, is an orally-administered, heterocyclic small molecule known to be an inhibitor of hypoxia inducible factor (HIF) prolyl hydroxylase. However, none of the studies have thus far thoroughly investigated its possible perturbations on membrane ion currents in endocrine or heart cells. In our studies, the whole-cell current recordings of the patch-clamp technique showed that the presence of roxadustat effectively and differentially suppressed the peak and late components of IK(DR) amplitude in response to membrane depolarization in pituitary tumor (GH3) cells with an IC50 value of 5.71 and 1.32 μM, respectively. The current inactivation of IK(DR) elicited by 10-sec membrane depolarization became raised in the presence of roxadustatt. When cells were exposed to either CoCl2 or deferoxamine (DFO), the IK(DR) elicited by membrane depolarization was not modified; however, nonactin, a K+-selective ionophore, in continued presence of roxadustat, attenuated roxadustat-mediated inhibition of the amplitude. The steady-state inactivation of IK(DR) could be constructed in the presence of roxadustat. Recovery of IK(DR) block by roxadustat (3 and 10 μM) could be fitted by a single exponential with 382 and 523 msec, respectively. The roxadustat addition slightly suppressed erg-mediated K+ or hyperpolarization-activated cation currents. This drug also decreased the peak amplitude of voltage-gated Na+ current with a slowing in inactivation rate of the current. Likewise, in H9c2 heart-derived cells, the addition of roxadustat suppressed IK(DR) amplitude in combination with the shortening in inactivation time course of the current. In high glucose-treated H9c2 cells, roxadustat-mediated inhibition of IK(DR) remained unchanged. Collectively, despite its suppression of HIF prolyl hydroxylase, inhibitory actions of roxadustat on different types of ionic currents possibly in a non-genomic fashion might provide another yet unidentified mechanism through which cellular functions are seriously perturbed, if similar findings occur in vivo.

Highlights

The hypoxia-inducible factor (HIF) pathway alters gene expression in response to low oxygen tension
The hypoxia inducible factor-1α (HIF-1α) expression in pituitary gland including pituitary tumor (GH3) cells has shown that it is closely linked to the occurrence or progression of pituitary adenoma [16,18,19,20,21,22,23,24]
We provide the evidence to unravel that roxadustat is able to modify membrane ionic currents in pituitary tumor (GH3) cells and in cardiac H9c2 cells, those actions of which are conceivably of clinical or pharmacological relevance and tend to be upstream of its inhibitory action on HIF prolyl hydroxylases

Summary

Introduction

The hypoxia-inducible factor (HIF) pathway alters gene expression in response to low oxygen tension. Roxadustat has been demonstrated to have therapeutic potential either for Parkinson’s diseases occurring in vitro or in vivo, for experimental retinal detachment, or for spinal cord injury, which are possibly through its regulation of redox reaction and mitochondrial function [5,12,13,14,15]. This compound could effectively exert neuroprotective effects against photoreceptor or neuronal cell death and prevent liver ischemia-reperfusion injury to donation after cardiac death [13,15,16]. The HIF-1α expression in pituitary gland including pituitary tumor (GH3) cells has shown that it is closely linked to the occurrence or progression of pituitary adenoma [16,18,19,20,21,22,23,24]

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Discussion

Conclusion