Biomedical subjects

K Meisheri

Publications and source records attributed to K Meisheri.

8 recordsLinked to original sources

PubMed2002

Preclinical studies characterizing the anti-migraine and cardiovascular effects of the selective 5-HT1D receptor agonist PNU-142633.

The present study describes the preclinical pharmacology of a highly selective 5-HT1D receptor agonist PNU-142633. PNU-142633 binds with a Ki of 6 nm at the human 5-HT1D receptor and a Ki of> 18 000 nm at the human 5-HT1B receptor. The intrinsic activity of PNU-142633 at the human 5-HT1D receptor was determined to be 70% that of 5-HT in a cytosensor cell-based assay compared with 84% for that of sumatriptan. PNU-142633 was equally effective as sumatriptan and a half-log more potent than sumatriptan in preventing plasma protein extravasation induced by electrical stimulation of the trigeminal ganglion. Like sumatriptan, PNU-142633 reduced the increase in cat nucleus trigeminal caudalis blood flow elicited by electrical stimulation of the trigeminal ganglion compared with the vehicle control. The direct vasoconstrictor potential of PNU-142633 was evaluated in vascular beds. Sumatriptan increased vascular resistance in carotid, meningeal and coronary arteries while PNU-142633 failed to alter resistance in these vascular beds. These data are discussed in relation to the clinical findings of PNU-142633 in a phase II acute migraine study.

Analgesics↗

PubMed1995

Receptor binding characterization in kidney membrane of [3H]U-37883, a novel ATP-sensitive K+ channel blocker with diuretic/natriuretic properties.

U-37883 (4-morpholinecarboximidine-N-1-adamantyl-N-cyclohexyl), a known blocker of ATP-sensitive K+ (KATP) channels, produces natriuresis/diuresis in vivo by a direct effect on the kidney. In the present study, the binding characteristics of the U-37883 receptor were investigated using pig kidney cortex microsomes. [3H]U-37883 (0.5-5 nM, 50 Ci/mmol) exhibited specific binding, which was reversible, increased linearly with protein concentration (50-500 micrograms/ml), and was destroyed after treatment with proteases. Scatchard plots derived from the competition experiments suggested the presence of a single class of low affinity binding sites, with a Kd of 225 nM and a Bmax of 7.8 pmol/mg of protein. A similar Kd value was derived from complementary studies dealing with association and dissociation kinetics. The binding of [3H]U-37883 was tissue specific, because very little specific binding could be detected in microsomes from rat insulinoma cells (RINm5F) and brain. In contrast, these membranes displayed high affinity specific binding of [3H]glyburide, another KATP channel blocker. Finally, analogs of U-37883 that were found to be active KATP channel blockers in isolated rabbit mesenteric artery and active in vivo as diuretics/natriuretics were also found to be active in displacing specific binding of [3H]U-37883, whereas the inactive analogs (no vascular KATP channel-blocking activity and no in vivo diuresis/natriuresis) were inactive in this binding assay. We suggest that the U-37883 binding site represents a functional receptor that mediates the KATP channel antagonism and natriuresis observed with this class of compounds.

Adamantane↗

PubMed1994

Functional receptors in Xenopus oocytes for U-37883A, a novel ATP-sensitive K+ channel blocker: comparison with rat insulinoma cells.

Follicle-enclosed Xenopus oocytes were used to describe the ATP-sensitive K+ (KATP) channel-blocking properties of U-37883A (4-morpholinecarboximidine-N-1-adamantyl-N'-cyclohexyl), in comparison with glibenclamide. In follicular oocytes, the KATP channel opener P1060 (30 microM), a pinacidil analog, activated a large outward K+ current that was blocked by glibenclamide (IC50 = 0.33 microM) and U-37883A (IC50 = 0.26 microM). P1060 activation was inhibited by both U-37883A and glibenclamide in a noncompetitive manner. U-37883A also blocked the KATP channel activation by cAMP (300 microM) and adenosine (10 microM). Single-channel studies on isolated follicular cells showed that U-37883A (10 microM) reduced the open probability of the KATP channel by 76%, without significantly modifying the single-channel current amplitude. Receptor binding studies with [3H]U-37883 in membranes from follicle-enclosed oocytes demonstrated a single class of low affinity binding sites, with a Kd of 450 nM and a Bmax of 17 pmol/mg of protein. Studies with analogs of U-37883A showed that U-52090A inhibited KATP current and displaced [3H]U-37883 from its binding site with similar potencies. In contrast, U-42069D neither inhibited KATP current nor competed with [3H]U-37883 binding. In RINm5F cells (an insulinoma cell line), U-37883A, unlike glibenclamide, failed to inhibit KATP current. Furthermore, there was no significant specific binding of [3H]U-37883 in RINm5F cell membranes, which displayed high levels of specific binding of [3H]glibenclamide. These data demonstrate the presence of a receptor for U-37883A-type guanidines that controls the activity of the endogenous KATP channels in follicle-enclosed oocytes. The available data collectively suggest that U-37883A is a more selective blocker of the follicular KATP channel, which is very similar to that in smooth muscle, than of the pancreatic beta cell KATP channel.

Adamantane↗

PubMed1984

Theoretical bases for vascular selectivity of Ca2+ antagonists.

The purpose of these studies was to characterize mechanisms of activation of vascular tissues in order to clarify possible theoretical bases for Ca2+-antagonist (CAt) selectivity. Activation of rabbit aorta, mesenteric artery, and mesenteric resistance vessels by agonists and depolarizing potassium (K+) solution, and inhibition by CAts were studied by measurement of contractions, 45Ca fluxes, and membrane potentials (via intracellular electrodes). We found that the CAts studied (D-600, diltiazem, and nisoldipine) inhibited K+-induced Ca2+ influx and contractions in a closely correlated manner, suggesting inhibition of Ca2+ entry as their primary, if not sole, mechanism of action. Diltiazem and nisoldipine had no effect on intracellular Ca2+ release or on the contractile protein system. The following evidence was obtained to support the tenet that norepinephrine (NE) and 80 mM K+ open two distinct Ca2+ channels, one receptor-operated (ROC) and the other potential-operated (POC): (a) NE activated the rabbit aorta without eliciting a change in membrane potential; (b) NE activation of the rabbit mesenteric resistance vessels was accompanied by membrane depolarization, but this depolarization was blocked by 10(-5) M diltiazem, whereas that induced by 80 mM K+ was not; (c) 45Ca influx stimulated by 80 mM K+ and that stimulated by a maximal [NE] were additive when the two agents were administered together in the aorta and the resistance vessels; (d) the Ca2+-channel agonist Bay K8644 opens the POC but not the ROC in rabbit aorta, as it is capable of stimulating Ca2+ influx in addition to 10(-5) M NE, but not to 80 mM K+; (e) the CAts show selective inhibition of the POC over the ROC in the aorta, whereas diltiazem preferentially inhibits the ROC in the resistance vessels; and (f) alpha adrenoreceptor occupation, phosphodiesterase inhibition, or dibutyryl cyclic AMP selectively inhibits the POC over the ROC in the aorta.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1982

Selectivity of calcium antagonistic action in vascular smooth muscle.

Various mechanisms can account for selective inhibition by calcium influx blocking drugs of smooth muscle contractile activity. Diltiazem inhibits calcium influx activated by membrane depolarization as well as alpha adrenergic receptor stimulation. Diltiazem does not inhibit intracellular calcium release nor cause net calcium extrusion up to concentrations of 10(-5) M. As with other calcium influx blocking agents, diltiazem is a potent inhibitor for modes of activation that depend entirely on the presence of extracellular calcium. The calcium influx was found to be mediated by at least three different pathways: one activated by depolarization and highly sensitive to D-600, a second activated by norepinephrine and less sensitive to D-600, and a third "passive leak," that is insensitive to D-600 and accounts for calcium permeability in the resting state. Molecular differences among these various calcium channels may provide the basis for specificity of action of the calcium influx blocking agents.

Animals↗

PubMed1982

Calcium fluxes in isolated rabbit aorta and guinea pig tenia coli.

Studies utilizing 45Ca have been helpful in analyzing the Ca control system in smooth muscle. Activation of rabbit aortic alpha receptors stimulates Ca influx and a release of Ca from superficial binding sites. Membrane depolarization by high K causes an influx, but no release. However, the influx pathways activated by alpha agonists and membrane depolarization are different and independent, because the maximal Ca influxes induced by each type of stimulation are additive. The cellular Ca pools that release Ca during pharmacological activation are shared by several agonists; release of cellular Ca by one agent after abolition of Ca influx inhibits Ca release by a second, different agonist. U44069, a stable prostaglandin H2 analog, induces both Ca influx and release. The extracellular Ca source for initial influx exchanges more slowly than free interstitial Ca, and may be located within the glycocalyx or on the outer membrane surface. An Na-Ca exchange process has been suggested as the important determinant of the transmembrane electrochemical Ca gradient. However, manipulation of the Na gradient by Na pump inhibition and Na substitution has provided data showing that Na-Ca exchange is a nonspecific process not directly involved in regulating [Ca]i. It is more likely that Ca extrusion is dependent on an ATPase in smooth muscle.

Animals↗

PubMed1980

Ca2+ movements in smooth muscle.

We describe the Ca2+ movements in smooth muscle cells at rest and during activation and relaxation as deduced from transplasmalemmal Ca2+ fluxes and contractile respnses. The general picture which emerges is: the resting cell has a [Ca2+]cyt below 10(-7) M and large gradients are poised across both the cell membrane and intracellular membranes. Excitation opens up Ca2+ channels which are linked to receptors and, if depolarization occurs, to other channels not linked to specific receptors but capable of sensing the membrane potential. Receptor activation also leads to release of Ca2+ from a limited intracellular Ca2+ pool which is superficially located because it has to be refilled from the outside. Relaxation is effected by Ca2+ accumulation by another intracellular Ca2+ pool, very likely sarcoplasmic reticulum, which does not release Ca2+ during activation. The sarcoplasmic reticulum Ca2+ pump can also decrease initial activation of the myofilaments. Elevation of cAMP levels may inhibit contraction by stimulating the sarcoplasmic reticulum Ca-ATPase. An enormous amount of research is still required to prove the above scheme and to localize and quantitate the various intracellular Ca2+ pools.

Animals↗

PubMedDate not supplied

Skinned coronary smooth muscle: calmodulin, calcium antagonists, and cAMP influence contractility.

The effects of Ca2+, calmodulin, cAMP, the catalytic subunit of cAMP-dependent protein kinase (CSU) and some Ca2+ antagonists were studied in chemically (Triton X-100) skinned coronary smooth muscle. Calmodulin increased the Ca2+ responsiveness of the muscle fiber as indicated by the reduction in the threshold as well as the half-maximal activating Ca2+ concentration. Trifluoroperazine, a calmodulin antagonist, inhibited Ca2+-calmodulin-induced contraction. Both cAMP and CSU were effective inhibitors of contraction induced at an intermediate Ca2+ concentration. Fendiline, a Ca2+-antagonist, at 2 x 10(-4) M produced a significant inhibitory effect, which was reduced by increasing the Ca2+ concentration. From other Ca2+ antagonists tested, W-7, but not D600 and verapamil, produced some inhibitory effect. The data indicate that the response of skinned coronary smooth muscle to Ca2+, calmodulin and cAMP are similar to those obtained with other skinned smooth muscles. Furthermore, skinned fiber preparation can serve as a useful tool to investigate possible direct effects of drugs on the activating and regulatory systems in smooth muscle.

Animals↗