The Prostacyclin Analogue, Treprostinil, Used in the Treatment of Pulmonary Arterial Hypertension, is a Potent Antagonist of TREK-1 and TREK-2 Potassium Channels

Pulmonary arterial hypertension (PAH) is an aggressive vascular remodeling disease that carries a high morbidity and mortality rate. Treprostinil (Remodulin) is a stable prostacyclin analogue with potent vasodilatory and anti-proliferative activity, approved by the FDA and WHO as a treatment for PAH. A limitation of this therapy is the severe subcutaneous site pain and other forms of pain experienced by some patients, which can lead to significant non-compliance. TWIK-related potassium channels (TREK-1 and TREK-2) are highly expressed in sensory neurons, where they play a role in regulating sensory neuron excitability. Downregulation, inhibition or mutation of these channels leads to enhanced pain sensitivity. Using whole-cell patch-clamp electrophysiological recordings, we show, for the first time, that treprostinil is a potent antagonist of human TREK-1 and TREK-2 channels but not of TASK-1 channels. An increase in TASK-1 channel current was observed with prolonged incubation, consistent with its therapeutic role in PAH. To investigate treprostinil-induced inhibition of TREK, site-directed mutagenesis of a number of amino acids, identified as important for the action of other regulatory compounds, was carried out. We found that a gain of function mutation of TREK-1 (Y284A) attenuated treprostinil inhibition, while a selective activator of TREK channels, BL-1249, overcame the inhibitory effect of treprostinil. Our data suggests that subcutaneous site pain experienced during treprostinil therapy may result from inhibition of TREK channels near the injection site and that pre-activation of these channels prior to treatment has the potential to alleviate this nociceptive activity

Characterization and regulation of wild‐type and mutant TASK‐1 two pore domain potassium channels indicated in pulmonary arterial hypertension

Key points The TASK-1 channel gene (KCNK3) has been identified as a possible disease-causing gene in heritable pulmonary arterial hypertension (PAH). In the present study, we show that novel mutated TASK-1 channels, seen in PAH patients, have a substantially reduced current compared to wild-type TASK-1 channels. These mutated TASK-1 channels are located at the plasma membrane to the same degree as wild-type TASK-1 channels. ONO-RS-082 and alkaline pH 8.4 both activate TASK-1 channels but do not recover current through mutant TASK-1 channels. We show that the guanylate cyclase activator, riociguat, a novel treatment for PAH, enhances current through TASK-1 channels but does not recover current through mutant TASK-1 channels. Pulmonary arterial hypertension (PAH) affects ∼15–50 people per million. KCNK3, the gene that encodes the two pore domain potassium channel TASK-1 (K2P3.1), has been identified as a possible disease-causing gene in heritable PAH. Recently, two new mutations have been identified in KCNK3 in PAH patients: G106R and L214R. The present study aimed to characterize the functional properties and regulation of wild-type (WT) and mutated TASK-1 channels and determine how these might contribute to PAH and its treatment. Currents through WT and mutated human TASK-1 channels transiently expressed in tsA201 cells were measured using whole-cell patch clamp electrophysiology. Localization of fluorescence-tagged channels was visualized using confocal microscopy and quantified with in-cell and on-cell westerns. G106R or L214R mutated channels were located at the plasma membrane to the same degree as WT channels; however, their current was markedly reduced compared to WT TASK-1 channels. Functional current through these mutated channels could not be restored using activators of WT TASK-1 channels (pH 8.4, ONO-RS-082). The guanylate cyclase activator, riociguat, enhanced current through WT TASK-1 channels; however, similar to the other activators investigated, riociguat did not have any effect on current through mutated TASK-1 channels. Thus, novel mutations in TASK-1 seen in PAH substantially alter the functional properties of these channels. Current through these channels could not be restored by activators of TASK-1 channels. Riociguat enhancement of current through TASK-1 channels could contribute to its therapeutic benefit in the treatment of PAH

The pi -> pi pi process in nuclei and the restoration of chiral symmetry

The results of an extensive campaign of measurements of the pi -> pi pi process in the nucleon and nuclei at intermediate energies are presented. The measurements were motivated by the study of strong pi pi correlations in nuclei. The analysis relies on the composite ratio C_{pi pi}^A, which accounts for the clear effect of the nuclear medium on the (pi pi) system. The comparison of the C_{pi pi}^A distributions for the (pi pi)_{I=J=0} and (pi pi)_{I=0,J=2} systems to the model predictions indicates that the C_{pi pi}^A behavior in proximity of the 2m_pi threshold is explainable through the partial restoration of chiral symmetry in nuclei.Comment: accepted for publication in Nucl. Phys.

General properties of the pion production reaction in nuclear matter

The pion production reaction $\pi^+ \to \pi^+\pi^{\pm}$ on $^{45}Sc$ was studied at incident pion energies of $T_{\pi^{+}}$ = 240, 260, 280, 300, and 320 MeV. The experiment was performed using the $M11$ pion-channel at TRIUMF, and multiparticle events, ($\pi^+,\pi^+\pi^{\pm}$) and ($\pi^+,\pi^+\pi^{\pm}p$), were detected with the CHAOS spectrometer. Results are reported in the form of both differential and total cross sections, and are compared to theoretical predictions and the reaction phase space. The present investigation of the T-dependence of the $\pi^+ A \to \pi^+\pi^{\pm} A'$ reaction complements earlier examinations of the A-dependence of the reaction, which was measured using $^{2}H$, $^{4}He$, $^{12}C$, $^{16}O$, $^{40}Ca$, and $^{208}Pb$ targets at $\sim$280 MeV. Some general properties of the pion-induced pion production reaction in nuclear matter will be presented, based on the combined results of the two studies.Comment: 23 pages, Latex, accepted for publication in Nucl. Phys.

The ππ\pi\pi interaction in nuclear matter from a study of the π+A→π+π±A′\pi^+ A \to \pi^+ \pi^{\pm} A' reactions

The pion-production reactions $\pi^+ A \to \pi^+\pi^{\pm} A'$ were studied on $^{2}H$, $^{12}C$, $^{40}Ca$, and $^{208}Pb$ nuclei at an incident pion energy of $T_{\pi^{+}}$=283 MeV. Pions were detected in coincidence using the CHAOS spectrometer. The experimental results are reduced to differential cross sections and compared to both theoretical predictions and the reaction phase space. The composite ratio $\cal C$$_{\pi\pi}^A$ between the $\pi^{+}\pi^{\pm}$ invariant masses on nuclei and on the nucleon is also presented. Near the $2m_{\pi}$ threshold pion pairs couple to $(\pi\pi)_{I=J=0}$ when produced in the $\pi^+\to \pi^+\pi^-$ reaction channel. There is a marked near-threshold enhancement of $\cal C$$_{\pi^+\pi^-}^A$ which is consistent with theoretical predictions addressing the partial restoration of chiral symmetry in nuclear matter. Furthermore, the behaviour of $\cal C$$_{\pi^+\pi^-}^A$ is well described when the restoration of chiral symmetry is combined with standard P-wave renormalization of pions in nuclear matter. On the other hand, nuclear matter only weakly influences $\cal C$$_{\pi^+\pi^+}^A$, which displays a flat behaviour throughout the energy range regardless of $A$.Comment: 30 pages, 16 figures, PS format, accepted for publication in Nucl. Phys

The Concise Guide to PHARMACOLOGY 2023/24: Ion channels.

The Concise Guide to PHARMACOLOGY 2023/24 is the sixth in this series of biennial publications. The Concise Guide provides concise overviews, mostly in tabular format, of the key properties of approximately 1800 drug targets, and over 6000 interactions with about 3900 ligands. There is an emphasis on selective pharmacology (where available), plus links to the open access knowledgebase source of drug targets and their ligands (https://www.guidetopharmacology.org/), which provides more detailed views of target and ligand properties. Although the Concise Guide constitutes almost 500 pages, the material presented is substantially reduced compared to information and links presented on the website. It provides a permanent, citable, point-in-time record that will survive database updates. The full contents of this section can be found at http://onlinelibrary.wiley.com/doi/10.1111/bph.16178. Ion channels are one of the six major pharmacological targets into which the Guide is divided, with the others being: G protein-coupled receptors, nuclear hormone receptors, catalytic receptors, enzymes and transporters. These are presented with nomenclature guidance and summary information on the best available pharmacological tools, alongside key references and suggestions for further reading. The landscape format of the Concise Guide is designed to facilitate comparison of related targets from material contemporary to mid-2023, and supersedes data presented in the 2021/22, 2019/20, 2017/18, 2015/16 and 2013/14 Concise Guides and previous Guides to Receptors and Channels. It is produced in close conjunction with the Nomenclature and Standards Committee of the International Union of Basic and Clinical Pharmacology (NC-IUPHAR), therefore, providing official IUPHAR classification and nomenclature for human drug targets, where appropriate

Two-Pore Domain Potassium Channels as Drug Targets: Anesthesia and Beyond.

Two-pore domain potassium (K2P) channels stabilize the resting membrane potential of both excitable and nonexcitable cells and, as such, are important regulators of cell activity. There are many conditions where pharmacological regulation of K2P channel activity would be of therapeutic benefit, including, but not limited to, atrial fibrillation, respiratory depression, pulmonary hypertension, neuropathic pain, migraine, depression, and some forms of cancer. Up until now, few if any selective pharmacological regulators of K2P channels have been available. However, recent publications of solved structures with small-molecule activators and inhibitors bound to TREK-1, TREK-2, and TASK-1 K2P channels have given insight into the pharmacophore requirements for compound binding to these sites. Together with the increasing availability of a number of novel, active, small-molecule compounds from K2P channel screening programs, these advances have opened up the possibility of rational activator and inhibitor design to selectively target K2P channels