Role of hERG potassium channel assays in drug development

Priest, Birgit T.; Bell, Ian M.; García, María L.

doi:10.4161/chan.2.2.6004

Cited by 110 publications

(81 citation statements)

References 66 publications

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“…Therefore, our data suggest that LDT3 and LDT5 are unlikely to cause the adverse effects associated with inhibition of important off-target receptors. We were also able to discard any interference of LDT3 and LDT5 (1 mM) with hERG K 1 channel function (data not shown), whose blockade can elicit potentially fatal cardiac arrhythmias (Priest et al, 2008), which is the reason why this test is absolutely required for new drug approval by regulatory authorities (Bowes et al, 2012;Peters 2013).…”

Section: Discussionmentioning

confidence: 99%

New Multi-target Antagonists of 1A-, 1D-Adrenoceptors and 5-HT1A Receptors Reduce Human Hyperplastic Prostate Cell Growth and the Increase of Intraurethral Pressure

Nascimento-Viana

Carvalho

Nasciutti

et al. 2015

Journal of Pharmacology and Experimental Therapeutics

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Benign prostatic hyperplasia (BPH) is characterized by stromal cell proliferation and contraction of the periurethral smooth muscle, causing lower urinary tract symptoms. Current BPH treatment, based on monotherapy with a 1A -adrenoceptor antagonists, is helpful for many patients, but insufficient for others, and recent reports suggest that stimulation of a 1D -adrenoceptors and 5-hydroxytryptamine (serotonin) (5-HT) 1A receptors contributes to cell proliferation. In this study, we investigated the potential of three N-phenylpiperazine derivatives (LDT3, LDT5, and LDT8) as multi-target antagonists of BPH-associated receptors. The affinity and efficacy of LDTs were estimated in isometric contraction and competition-binding assays using tissues (prostate and aorta) and brain membrane samples enriched in specific on-or off-target receptors. LDTs' potency was estimated in intracellular Ca 21 elevation assays using cells overexpressing human a 1 -adrenoceptor subtypes. The antiproliferative effect of LDTs on prostate cells from BPH patients was evaluated by viable cell counting and 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide assays. We also determined LDTs' effects on rat intraurethral and arterial pressure. LDT3 and LDT5 are potent antagonists of a 1A -, a 1D -adrenoceptors, and 5-HT 1A receptors (K i values in the nanomolar range), and fully inhibited phenylephrine-and 5-HT-induced proliferation of BPH cells. In vivo, LDT3 and LDT5 fully blocked the increase of intraurethral pressure (IUP) induced by phenylephrine at doses (ED 50 of 0.15 and 0.09 mg.kg 21 , respectively) without effect on basal mean blood pressure. LDT3 and LDT5 are multi-target antagonists of key receptors in BPH, and are capable of triggering both prostate muscle relaxation and human hyperplastic prostate cell growth inhibition in vitro. Thus, LDT3 and LDT5 represent potential new lead compounds for BPH treatment.

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Section: Discussionmentioning

confidence: 99%

New Multi-target Antagonists of 1A-, 1D-Adrenoceptors and 5-HT1A Receptors Reduce Human Hyperplastic Prostate Cell Growth and the Increase of Intraurethral Pressure

Nascimento-Viana

Carvalho

Nasciutti

et al. 2015

Journal of Pharmacology and Experimental Therapeutics

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“…Our ability to predict which small molecules will block hERG is poor and therefore all compounds under consideration for drug development are tested in a hERG activity assay. Several assays exist: cell-based functional assays, such as thallium and/or rubidium flux, nonfunctional assays, such as ligand binding and/or fluorescence polarization, and electrophysiology (22,23,31,32). The gold standard is electrophysiology, but it can be slow and expensive when the need exists for evaluating many compounds.…”

Section: Significancementioning

confidence: 99%

“…None of the methods currently used-electrophysiology (17,18), cell-based assays such as Tl + flux assays/membrane potential dye assays/yeast growth assays (19)(20)(21), and binding/inhibitor competition assays (22,23)fulfill all three of the above criteria. We developed and present here an assay that does fulfill these criteria.…”

mentioning

confidence: 99%

Novel cell-free high-throughput screening method for pharmacological tools targeting K ⁺ channels

Brown

Wang

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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K+ channels, a superfamily of ∼80 members, control cell excitability, ion homeostasis, and many forms of cell signaling. Their malfunctions cause numerous diseases including neuronal disorders, cardiac arrhythmia, diabetes, and asthma. Here we present a novel liposome flux assay (LFA) that is applicable to most K+ channels. It is robust, low cost, and high throughput. Using LFA, we performed small molecule screens on three different K+ channels and identified new activators and inhibitors for biological research on channel function and for medicinal development. We further engineered a hERG (human ether-à-go-go-related gene) channel, which, when used in LFA, provides a highly sensitive (zero false negatives on 50 hERG-sensitive drugs) and highly specific (zero false positives on 50 hERG-insensitive drugs), low-cost hERG safety assay.

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“…8 In addition, discrepancies between automated and manual electrophysiology in the potencies of compounds have been observed that are thought to be the result of hydrophobic compounds absorbing to the surface of automated patch clamp plates. 8,9 Attempts to mitigate these problems increase the cost and/or the time involved in the evaluation of compounds.…”

Section: Introductionmentioning

confidence: 98%

A Pharmacologically Validated, High-Capacity, Functional Thallium Flux Assay for the Human Ether-à-go-go Related Gene Potassium Channel

Schmalhofer

Swensen

Thomas

et al. 2010

ASSAY and Drug Development Technologies

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The voltage-gated potassium channel, human Ether-à-go-go related gene (hERG), represents the molecular component of IKr, one of the potassium currents involved in cardiac action potential repolarization. Inhibition of IKr increases the duration of the ventricular action potential, reflected as a prolongation of the QT interval in the electrocardiogram, and increases the risk for potentially fatal ventricular arrhythmias. Because hERG is an appropriate surrogate for IKr, hERG assays that can identify potential safety liabilities of compounds during lead identification and optimization have been implemented. Although the gold standard for hERG evaluation is electrophysiology, this technique, even with the medium capacity, automated instruments that are currently available, does not meet the throughput demands for supporting typical medicinal chemistry efforts in the pharmaceutical environment. Assays that could provide reliable molecular pharmacology data, while operating in high capacity mode, are therefore desirable. In the present study, we describe a high-capacity, 384- and 1,536-well plate, functional thallium flux assay for the hERG channel that fulfills these criteria. This assay was optimized and validated using different structural classes of hERG inhibitors. An excellent correlation was found between the potency of these agents in the thallium flux assay and in electrophysiological recordings of channel activity using the QPatch automated patch platform. Extension of this study to include 991 medicinal chemistry compounds from different internal drug development programs indicated that the thallium flux assay was a good predictor of in vitro hERG activity. These data suggest that the hERG thallium flux assay can play an important role in supporting drug development efforts.

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Role of hERG potassium channel assays in drug development

Cited by 110 publications

References 66 publications

New Multi-target Antagonists of 1A-, 1D-Adrenoceptors and 5-HT1A Receptors Reduce Human Hyperplastic Prostate Cell Growth and the Increase of Intraurethral Pressure

New Multi-target Antagonists of 1A-, 1D-Adrenoceptors and 5-HT1A Receptors Reduce Human Hyperplastic Prostate Cell Growth and the Increase of Intraurethral Pressure

Novel cell-free high-throughput screening method for pharmacological tools targeting K ⁺ channels

A Pharmacologically Validated, High-Capacity, Functional Thallium Flux Assay for the Human Ether-à-go-go Related Gene Potassium Channel

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Role of hERG potassium channel assays in drug development

Cited by 110 publications

References 66 publications

New Multi-target Antagonists of 1A-, 1D-Adrenoceptors and 5-HT1A Receptors Reduce Human Hyperplastic Prostate Cell Growth and the Increase of Intraurethral Pressure

New Multi-target Antagonists of 1A-, 1D-Adrenoceptors and 5-HT1A Receptors Reduce Human Hyperplastic Prostate Cell Growth and the Increase of Intraurethral Pressure

Novel cell-free high-throughput screening method for pharmacological tools targeting K + channels

A Pharmacologically Validated, High-Capacity, Functional Thallium Flux Assay for the Human Ether-à-go-go Related Gene Potassium Channel

Contact Info

Product

Resources

About

Novel cell-free high-throughput screening method for pharmacological tools targeting K ⁺ channels