Hydroxypropyl beta-cyclodextrins: a misleading vehicle for the in vitro hERG current assay.
Delayed cardiac repolarization and fatal proarrhythmia have been linked to block of the repolarizing current, Ikr or hERG (human ether-a-go-go related gene) current. Thus, determining the potency of hERG block is critical in evaluating cardiac safety during preclinical development. Hydroxypropyl beta-cyclodextrins (HbetaC) are cyclic oligosaccharides used to enhance drug solubility. To evaluate the utility of HbetaC to enhance drug solubility in hERG screening assays, we studied the effect of HbetaC on hERG current and the sensitivity of the hERG assay to 3 structurally different hERG blocking drugs using whole-cell voltage clamp technique and HEK-293 cells expressing the hERG channel. HbetaC inhibited hERG activation and tail current and accelerated current deactivation in a concentration-dependent manner. HbetaC (6%) reduced the apparent potency of block by terfenadine (IC50 12000 nM vs 45 nM), cisapride (IC50 281 nM vs 28 nM), and E-4031 (163 nM vs 26 nM). Reduced potency of block was consistent with loss of activity as a result of complexation with HbetaC by terfenadine and cisapride (demonstrated in solubility studies) and interactions with HbetaC by E-4031 (demonstrated in absorbance studies). These results demonstrate that HbetaC is an unsuitable agent for enhancing compound solubility in the in vitro hERG current assay and may mask drug effects, allowing potentially dangerous drugs to advance into clinical development.
Mikhail
Ann
A
Department of Integrative Pharmacology, Abbott, Abbott Park, Illinois 60064, USA.
Fischer
Cristina
C
Patel
Ameesha
A
Long
Michelle A
MA
Limberis
James T
JT
Martin
Ruth L
RL
Cox
Bryan F
BF
Gintant
Gary A
GA
Su
Zhi
Z
D000073738:2-Hydroxypropyl-beta-cyclodextrin; D000889:Anti-Arrhythmia Agents / Q000494:pharmacology; D001145:Arrhythmias, Cardiac / Q000139:chemically induced / Q000503:physiopathology; D001681:Biological Assay; D020117:Cisapride / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D051638:Ether-A-Go-Go Potassium Channels / Q000187:drug effects*; D005079:Excipients / Q000494:pharmacology*; D005765:Gastrointestinal Agents / Q000494:pharmacology; D006634:Histamine H1 Antagonists / Q000009:adverse effects / Q000494:pharmacology; D006801:Humans; D020128:Inhibitory Concentration 50; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000502:physiology; D010880:Piperidines / Q000494:pharmacology; D026902:Potassium Channel Blockers / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D012995:Solubility / Q000187:drug effects; D016593:Terfenadine / Q000009:adverse effects / Q000494:pharmacology; D013997:Time Factors; D047392:beta-Cyclodextrins / Q000494:pharmacology*
D000889:Anti-Arrhythmia Agents; D051638:Ether-A-Go-Go Potassium Channels; D005079:Excipients; D005765:Gastrointestinal Agents; D006634:Histamine H1 Antagonists; D010880:Piperidines; D026902:Potassium Channel Blockers; D011725:Pyridines; D047392:beta-Cyclodextrins; C063968:E 4031; D000073738:2-Hydroxypropyl-beta-cyclodextrin; D016593:Terfenadine; D020117:Cisapride