Inhibition of the bioenergetic functions of isolated rat liver mitochondria by polyamines

The abilities of the naturally occuring polyamines, putrescine, spermidine and spermine, to affect variables related to the bioenergetic functions of isolated rat liver mitochondria were studied. At concentrations comparable to those present intracellularly, the polyamines inhibited state 4 respiration, but they had much less effect on state 3 or uncoupled respiration. The concentrations required to produce 25% inhibition (I₂₅) of state 4 respiration varied according to the polyamine, with putrescine being least effective (I₂₅, 20mM) and spermidine and spermine being more effective and comparable (I₂₅, 7.5 and 7.0mM respectively). This inhibition was antagonized by 15 mM potassium and enhanced by valinomycin and 4 mM magnesium. Inhibition of monoamine oxidase, an enzyme of outer mitochondrial membrane, was also observed to occur. Addition of polyamines to mitochondrial suspensions caused an increase in the optical density and protected against the swelling effects of sublytic concentrations of Triton X-100. By electron microscopy, polyamines were found to cause the outer mitochondrial compartment to collapse bringing the inner and outer membranes into apparent contact with one another. The electrophoretic mobility of mitochondria toward the anode was markedly slowed by polyamines (i.e. 50% by 1.25 mM spermine), indicating surface binding and neutralization of the negative surface charge. In almost all of the above mitochondrial effects, spermine and spermidine were similar in effectiveness and putrescine was less effective. It is suggested that polyamines may be capable of modulating respiration of isolated mitochondria by binding to non-specific anionic sites at the surface of the inner mitochondrial membrane. Neutralization of the net negative surface potential may interfere with cation fluxes across the membrane, particularly those of potassium.     

Uptake Mechanism of Trientine by Rat Intestinal Brush-border Membrane Vesicles

The uptake characteristics of trientine by rat intestinal brush-border membrane vesicles were studied. The uptake characteristics of trientine were similar to those of the physiological polyamines with respect to the excessive accumulation in vesicles, the pH dependency, the temperature dependency and the ineffectiveness of K⁺ diffusion potential (inside negative). The initial uptake of trientine was saturable with a K_m value of 1.13 mM, which was larger than that of spermine and spermidine. Furthermore, the uptake rate of trientine was dose-dependently inhibited by spermine and spermidine. Spermine competitively inhibited the uptake of trientine with a K_i value of 18.6 μM, and it was close to the K_m value for spermine (30.4 μM). These data suggested that the uptake of trientine was similar to that of spermine and spermidine in rat small intestinal brush-border membrane vesicles, and these polyamines seem to inhibit the absorption of trientine from the gastrointestinal tract.     

Characterisation of [3H]gabapentin binding to a novel site in rat brain: homogenate binding studies

The binding characteristics of [³H]gabapentin, the radiolabelled analogue of the novel anticonvulsant gabapentin (1-(aminomethyl)cyclohexaneacetic acid) were studied using purified synaptic plasma membranes prepared from rat cerebral cortex. In 10 mM HEPES buffer [³H]gabapentin bound to a single population of sites with high affinity (K_D = 38 ± 2.8 mM) with a maximum binding capacity of 4.6 ± 0.4 pmol/mg protein, reaching equilibrium after 30 min at 20°C. This novel site was unique to the central nervous system with little or no specific [³H]gabapentin binding being measurable in a range of peripheral tissues. Binding was potently inhibited by a range of gabapentin analogues and 3-alkyl substituted γ-aminobutyric acid (GABA) derivatives although GABA itself and the selective GABA_B receptor ligand baclofen, were only weakly active. Gabapentin itself (IC₅₀ = 80 nM) and 3-isobutyl GABA (IC₅₀ = 80 nM) which also has anticonvulsant properties, showed the highest affinity for the binding site. Of a wide range of other pharmacologically active compounds only the polyamines spermine and spermidine influenced [³H]gabapentin binding, with both compounds producing a maximum of 50% inhibition of specific binding. Magnesium ions produced a similar pattern of inhibition but the effect of the polyamines and magnesium ions were not additive. The data provide evidence for the existence in brain of a novel binding site that may mediate the anticonvulsant effects of gabapentin and other potential anticonvulsant compounds.     

The accumulation of diamines and polyamines into rat lung slices

The diamine cadaverine, and the polyamines spermidine and spermine have been shown to accumulate into rat lung slices by an uptake process which obeyed saturation kinetics. The apparent K_m values for the accumulation process of cadaverine, spermidine and spermine were 19, 11 and 15 μM respectively with V_max values of 937, 768 and 617 nmoles/g wet weight/hr respectively. The accumulation was KCN sensitive, indicative of an energy dependent process, although spermine did show some non-specific binding to lung tissue. Cadaverine, spermidine and spermine were not accumulated by slices of liver, kidney, heart and spleen to concentrations much greater than that in the medium. They were accumulated, however, by a KCN sensitive process into brain slices although the accumulation was much less than that which occurred in lung slices. The diamine, putrescine, exhibited a concentration-dependent inhibition of the ability of lung slices to accumulate cadaverine and the polyamines. These data have led us to conclude that the transport process in the lung, which has recently been shown to accumulate the diamine putrescine, is also capable of accumulating cadaverine, spermidine and spermine. Thus, by analogy with putrescine, there exists in specific lung cells a membrane receptor(s) which is selective in its acceptance and transport of diamines and polyamines.     

Polyamines and Their Metabolites as Diagnostic Markers of Human Diseases

Polyamines, putrescine, spermidine and spermine, are ubiquitous in living cells and are essential for eukaryotic cell growth. These polycations interact with negatively charged molecules such as DNA, RNA, acidic proteins and phospholipids and modulate various cellular functions including macromolecular synthesis. Dysregulation of the polyamine pathway leads to pathological conditions including cancer, inflammation, stroke, renal failure and diabetes. Increase in polyamines and polyamine synthesis enzymes is often associated with tumor growth, and urinary and plasma contents of polyamines and their metabolites have been investigated as diagnostic markers for cancers. Of these, diacetylated derivatives of spermidine and spermine are elevated in the urine of cancer patients and present potential markers for early detection. Enhanced catabolism of cellular polyamines by polyamine oxidases (PAO), spermine oxidase (SMO) or acetylpolyamine oxidase (AcPAO), increases cellular oxidative stress and generates hydrogen peroxide and a reactive toxic metabolite, acrolein, which covalently incorporates into lysine residues of cellular proteins. Levels of protein-conjuagated acrolein (PC-Acro) and polyamine oxidizing enzymes were increased in the locus of brain infarction and in plasma in a mouse model of stroke and also in the plasma of stroke patients. When the combined measurements of PC-Acro, interleukin 6 (IL-6), and C-reactive protein (CRP) were evaluated, even silent brain infarction (SBI) was detected with high sensitivity and specificity. Considering that there are no reliable biochemical markers for early stage of stroke, PC-Acro and PAOs present promising markers. Thus the polyamine metabolites in plasma or urine provide useful tools in early diagnosis of cancer and stroke.     

Polyamines allosterically modulate [3H]nitrendipine binding to the voltage-sensitive calcium channel in rat brain

The effects of polyamines on radioligand binding to the slow voltage-dependent Ca²⁺ channel were studied using membranes from the rat cerebral cortex. [³H]Diltiazem binding was inhibited by arcaine (IC₅₀ = 55 μM) and, in decreasing order of potency, by agmatine, spermidine, spermine and putrescine. Under control conditions, only spermidine and spermine allosterically inhibited [³H]nitrendipine binding while arcaine, agmatine and putrescine were inactive. Nevertheless, putrescine antagonized the effect of spermine as well as the allosteric effects of diltiazem and verapamil on the binding of [³H]nitrendipine, in a manner analogous to that shown previously for Ca²⁺. Thus, polyamines may function as endogenous modulators of the voltage-dependent Ca²⁺ channel.     

Uptake and disposition of putrescine, spermidine, and spermine by rabbit lungs

The pulmonary uptake and accumulation of the three polyamines in intact, ventilated, and perfused rabbit lungs was investigated. Lungs were perfused using Krebs-Ringer bicarbonate buffer with albumin in which putrescine, spermidine, or spermine were included at an initial concentration of 10(-3), 10(-2), 10(-1), 5, 10, or 20 mM. At a 5 mM concentration of spermidine and spermine, the uptake by isolated lungs reached a steady state equilibrium in 20-30 min of perfusion. This did not occur for putrescine, which showed linear uptake throughout the entire period of the 60-min perfusion. The lung uptake of putrescine for all perfusate concentrations was greater than that of spermidine or spermine, but all three showed concentration-dependent linear uptake. In the presence of harmaline (1 mM) and ouabain (1 mM), isolated perfused rabbit lungs showed a decrease in uptake of putrescine although no effect was seen for spermidine and spermine. Perfusate containing decreased sodium showed no effect on putrescine uptake by isolated rabbit lungs, but there was a significant increase in the uptake of spermidine and spermine. Significant uptake of all three polyamines was also observed when incubated separately with rabbit lung slices for 60 min. HPLC analysis of lung, the perfusate samples, lung slices, and the incubation medium after a 60-min incubation did not indicate the presence of metabolites of these polyamines. Likewise, the analysis of the lung homogenate incubated with polyamines did not show any metabolites confirming the absence of detectable pulmonary metabolism.(ABSTRACT TRUNCATED AT 250 WORDS)     

Stimulation and inhibition of 1,3-bis(2-chloroethyl)-1-nitrosourea-induced strand breaks and interstrand cross-linking in Col E1 plasmid deoxyribonucleic acid by polyamines and inorganic cations

The influence of various polyamines and metallic cations on 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU)-induced DNA single-strand breaks and DNA interstrand cross-linking was in Col E1 plasmid using electrophoretic techniques. Spermidine and spermine (0.4 to 1.5 mM concentration range) markedly stimulated BCNU-induced DNA nicking, whereas putrescine had no effect on the nicking process. In contrast to the polyamines, BCNU-induced DNA nicking was decreased by the three inorganic cations, Na+ (100 and 200 mM), Mg2+ (0.5 and 1.5 mM), and Co3+ (NH3)6 (0.2 and 0.4 mM), with the trivalent hexamminecobalt ions being most inhibitory. When the monofunctional N-methyl-N-nitrosourea (MNU) was used (instead of the bifunctionally active BCNU) to alkylate Col E1 DNA, nicking of the DNA was inhibited by spermidine. Furthermore, the ability of chloroethylated Col E1 DNA to form interstrand cross-links after treatment with BCU was inhibited by 0.5 mM spermidine and 0.5 mM spermine, both concentrations within the intracellular range. Putrescine at 3-6 mM only marginally stimulated DNA cross-linking. In comparison, the inorganic cations all enhanced Col E1 DNA cross-linking by BCNU, with the rank order of cross-link stimulation being Mg2+, Na+, and Co3+ (NH3)6. These results provide evidence that polyamines can interact with DNA to modulate chloroethylnitrosourea-induced DNA damage and that the interaction is not only a function of the charge on the polyamine molecule but also of the chemical structure of the polyamine.     

Relaxant Effect of Spermidine on Acethylcholine and High K+-induced Gastric Contractions of Guinea-Pig

In our previous study, we found that spermine and putrescine inhibited spontaneous and acetylcholine (ACh)-induced contractions of guinea-pig stomach via inhibition of L-type voltage-dependent calcium current (VDCC_L). In this study, we also studied the effect of spermidine on mechanical contractions and calcium channel current (I_Ba), and then compared its effects to those by spermine and putrescine. Spermidine inhibited spontaneous contraction of the gastric smooth muscle in a concentration-dependent manner (IC₅₀=1.1±0.11 mM). Relationship between inhibition of contraction and calcium current by spermidine was studied using 50 mM high K⁺-induced contraction: Spermidine (5 mM) significantly reduced high K⁺ (50 mM)-induced contraction to 37±4.7% of the control (p<0.05), and inhibitory effect of spermidine on I_Ba was also observed at a wide range of test potential in current/voltage (I/V) relationship. Pre- and post-application of spermidine (5 mM) also significantly inhibited carbachol (CCh) and ACh-induced initial and phasic contractions. Finally, caffeine (10 mM)-induced contraction which is activated by Ca²⁺-induced Ca²⁺ release (CICR),'' was also inhibited by pretreatment of spermidine (5 mM). These findings suggest that spermidine inhibits spontaneous and CCh-induced contraction via inhibition of VDCC_L and Ca²⁺ releasing mechanism in guinea-pig stomach.     

The natural polyamines spermidine and spermine prevent bone loss through preferential disruption of osteoclastic activation in ovariectomized mice

BACKGROUND AND PURPOSE

Although naturally occurring polyamines are indispensable for a variety of cellular events in eukaryotic cells, little attention has been paid to their physiological and pathological significance in bone remodelling to date. In this study, we evaluated the pharmacological properties of several natural polyamines on the functionality and integrity of bone in both in vitro and in vivo experiments.

EXPERIMENTAL APPROACH

Mice were subjected to ovariectomy (OVX) and subsequent oral supplementation with either spermidine or spermine for determination of the bone volume together with different parameters regarding bone formation and resorption by histomorphometric analyses in vivo. Pre-osteoclasts were cultured with receptor activator of NF-κB ligand (RANKL), with or without spermidine and spermine to determine cellular maturation by tartrate-resistant acid phosphatase (TRAP) staining and cellular viability by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide reduction in vitro.

KEY RESULTS

Spermidine or spermine, given in drinking water for 28 days, significantly prevented the increased osteoclast surface/bone surface ratio and the reduced bone volume following OVX in mice. Either spermidine or spermine significantly inhibited the increased number of multinucleated TRAP-positive cells in osteoclasts cultured with RANKL in a concentration-dependent manner without affecting cell survival.

CONCLUSIONS AND IMPLICATIONS

The natural polyamines spermidine and spermine prevented OVX-induced bone loss through the disruption of differentiation and maturation of osteoclasts, rather than affecting osteoblasts. The supplementation with these natural polyamines could be beneficial for the prophylaxis as well as therapy of metabolic bone diseases such as post-menopausal osteoporosis.     

Spermine-induced negative inotropic effect in isolated rat heart,is mediated through the release of ATP

Putrescine, spermidine and spermine are natural compounds found in up to millimolar concentrations in eukaryotic and prokaryotic cells. At physiologic pH, the polyamines are protonated (+2, +3 and +4 charges), their polycationic properties lead to the assumption that they could affect physiological systems by binding to anionic sites of the cellular membrane and/or by modulating ion channels. At the cardiovascular level, their effects are not completely understood. However, these compounds may be able to exert the induction of synthesis and release of cellular mediators. In an attempt to explore this possibility, we used the isolated and perfused rat heart, Langendorff, model in order to evaluate the inotropic effects of these polyamines, putrescine, spermidine and spermine. Dose-response curves (0.1-0.6 mM) for putrescine, spermidine and spermine were constructed; with the finding that spermine had the largest negative effect. The obtained effects were not blocked by nitric oxide synthesis inhibitors (L-NAME), H(1) and H(2) receptor antagonists (Brompheniramine and Cimetidine) or by Glibenclamide, an antagonist of ATP-sensitive K(+) channels. We found that spermine-induced and increased ATP concentration in cardiac effluents. Reactive Blue, a P(2y) purinoreceptor antagonist and Aminophylline, an unspecific adenosine receptor antagonist, blocked the spermine-induced effects. These results showed that ATP, at least in part, is responsible of the spermine cardiovascular effects. Adenosine was shown to also play an important role on those effects.     

Effects of benzodiazepines and valproic acid on brain aldehyde reductase and a proposed mechanism of anticonvulsant action

Benzodiazepines (clonazepam, diazepam, flurazepam, fosazepam, lorazepam, nitrazepam, oxazepam and R07-5205) were shown to inhibit the activity of brain aldehyde reductase obtained from DBA/2J mice with the ic₅₀ values (concentration of inhibitor at 50 per cent of control activity) ranging from 0.24 to 7.0 mM. ed₅₀ Values of these benzodiazepines for protection against maximal electroshock-induced convulsions were determined for DBA/2J mice which were pretreated with either saline or β-diethylaminoethyl diphenylpropylacetate (SKF-525A), an inhibitor of microsomal drug-metabolizing systems. Spearman rank order and Pearson correlation coefficients between the ic₅₀ values for inhibition of aldehyde reductase activity and the ed₅₀ values for protection against maximal electroshock-induced convulsions were calculated to be 0.62 and 0.82, respectively, for a group of eight benzodiazepines. When the animals were pretreated with SKF-525A, the correlation coefficients were 0.83 and 0.71, respectively. R_m values, indicators of relative lipid solubility, were measured for these benzodiazepines. Correlations between R_m values and ic₅₀ values or ed₅₀ values were not significant at the 95 per cent confidence level.Valproic acid inhibited DBA/2J mouse brain aldehyde reductase activity with an ic₅₀ value of 7 × 10^?5 M. Data presented in this study are consistent with the hypothesis that highly reactive aldehyde intermediates of biogenic amine metabolism may be implicated in anticonvulsant drug action.     

Effects of spermine on the relaxation response of rat detrusor smooth muscles

Endogenous polyamines are known to influence excitation-contraction coupling in smooth muscle. This study was designed to determine the effects of the polyamines spermine, spermidine, and putrescine on the contractile responses of rat detrusor smooth muscles. Under physiological conditions, isometric tension recordings were made of isolated bladder strips from excised rat bladder. The effects of spermine, spermidine, and putrescine (1 mM each) on the bladder contractions induced by various agents, i.e., acetylcholine, bethanechol, high-K, and tetraethylammonium (TEA) were measured. A conventional patch clamp technique was used in whole cell mode with single smooth muscle cells of rat bladder. Calcium channel currents were recorded to determine the effects of spermine on channel activities. Polyamines elicited a concentration-dependent relaxations on the contractile agents induced contractures. Spermine showed the most potent relaxation effect of the polyamines examined, and relaxed the contractions induced by the agents. Calcium channel activities were significantly reduced by adding 1 mM spermine to the bath. We concluded that spermine exerts a potent relaxant effect on rat bladder smooth muscle, and this effect appears to be mediated by calcium channel antagonism.     

Physiological functions of polyamines and regulation of polyamine content in cells

Polyamines (putrescine, spermidine, and spermine) are essential for normal cell growth. The polyamine level in cells is regulated by biosynthesis, degradation, and transport. The role of antizyme on polyamine biosynthesis and transport in mammalian cells and characteristics of polyamine transport in Escherichia coli and yeast are described briefly in this review. In addition, the effects of polyamines on protein synthesis and the NMDA receptor are outlined. Finally, the correlation between acrolein produced from polyamines by polyamine oxidase and chronic renal failure and brain stroke is summarized. Increased levels of polyamine oxidase and acrolein are good markers of chronic renal failure and brain stroke.     

Polyamines allosterically modulate [3H]nitrendipine binding to the voltage-sensitive calcium channel in rat brain.

The effects of polyamines on radioligand binding to the slow voltage-dependent Ca2+ channel were studied using membranes from the rat cerebral cortex. [3H]Diltiazem binding was inhibited by arcaine (IC50 = 55 microM) and, in decreasing order of potency, by agmatine, spermidine, spermine and putrescine. Under control conditions, only spermidine and spermine allosterically inhibited [3H]nitrendipine binding while arcaine, agmatine and putrescine were inactive. Nevertheless, putrescine antagonized the effect of spermine as well as the allosteric effects of diltiazem and verapamil on the binding of [3H]nitrendipine, in a manner analogous to that shown previously for Ca2+. Thus, polyamines may function as endogenous modulators of the voltage-dependent Ca2+ channel.     

Effects of 2,4-dichlorophenoxyacetic acid on polyamine synthesis in Chinese hamster ovary cells

It was found that 1 mM 2,4-dichlorophenoxyacetate (2,4-D) inhibited DNA and protein synthesis in Chinese hamster ovary (CHO) cells. When the possible relationship of this phenomenon to the presence of polyamines in the culture medium was investigated, it was found that: (a) the pesticide inhibited ornithine decarboxylase activity; (b) when the concentration of polyamines present in cells treated with the pesticide was determined, the putrescine concentration did not change, and the spermine and spermidine concentration decreased; (c) the addition of spermidine and spermine to CHO cells grown in the presence of 2,4-D normalized DNA and protein synthesis. Putrescine did not have any effect.     

Hyperglycaemia produced by the polyamines spermine and spermidine

1 Intravenous injection of rabbits with the polyamines spermine (10-30 mg/kg) and spermidine (50-90 mg/kg) produced hyperglycaemia.2 Given by intraventricular injection, spermine and spermidine gave rise to hyperglycaemia in doses 250-500 times smaller than those effective intravenously.3 Intraventricular injections of the polyamines also produced hyperglycaemia in rats. Adrenal demedullation abolished the response.4 In the rabbit, the hyperglycaemia resulting from intraventricular injection of polyamines was abolished in reserpine-treated animals. Anaesthesia had no effect on the response unless accompanied by anoxia, when the response was potentiated.5 Intracisternal injections of the polyamines in rabbits were less effective than intraventricular injections in that doses about four times larger were required to elicit an equivalent response.     

Contribution of peripheral vanilloid receptor to the nociception induced by injection of spermine in mice

Polyamines (putrescine, spermidine and spermine) are important endogenous regulators of ion channels, such as vanilloid (TRPV1), glutamatergic (NMDA or AMPA/kainate) and acid-sensitive (ASIC) receptors. In the present study, we have investigated the possible nociceptive effect induced by polyamines and the mechanisms involved in this nociception in vivo. The subcutaneous (s.c.) injection of capsaicin (as positive control), spermine, spermidine or putrescine produced nociception with ED₅₀ of 0.16 (0.07-0.39) nmol/paw, 0.4 (0.2-0.7) μmol/paw, 0.3 (0.1-0.9) μmol/paw and 3.2 (0.9-11.5) μmol/paw, respectively. The antagonists of NMDA (MK801, 1 nmol/paw), AMPA/kainate (DNQX, 1 nmol/paw) or ASIC receptors (amiloride, 100 nmol/paw) failed to reduce the spermine-trigged nociception. However, the TRPV1 antagonists capsazepine or SB366791 (1 nmol/paw) reduced spermine-induced nociception, with inhibition of 81 ± 10 and 68 ± 9%, respectively. The previous desensitization with resiniferatoxin (RTX) largely reduced the spermine-induced nociception and TRPV1 expression in the sciatic nerve, with reductions of 82 ± 9% and 67 ± 11%, respectively. Furthermore, the combination of spermine (100 nmol/paw) and RTX (0.005 fmol/paw), in doses which alone were not capable of inducing nociception, produced nociceptive behaviors. Moreover, different concentrations of spermine (3-300 μM) enhanced the specific binding of [³H]-RTX to TRPV1 receptor. Altogether, polyamines produce spontaneous nociceptive effect through the stimulation of TRPV1, but not of ionotropic glutamate or ASIC receptors.     

alpha-Methyl polyamines: metabolically stable spermidine and spermine mimics capable of supporting growth in cells depleted of polyamines.

In order to assess the tolerance of the target enzyme spermine synthase for alpha-substituents on the aminopropyl moiety of the substrate spermidine, 1-methylspermidine (MeSpd, 2) was synthesized. It was determined that MeSpd is a poor substrate for spermine synthase and is not a substrate for spermidine N1-acetyltransferase, suggesting that alpha-methylated polyamines might be metabolically stable and therefore useful tools for studying polyamine effects in intact cells. On the basis of initial cellular results with 2, 1-methylspermine (MeSpm, 3) and 1,12-dimethylspermine (Me2Spm, 4) were also synthesized. When added to cells (L1210, SV-3T3, or HT29) depleted of both putrescine and spermidine by prior treatment with alpha-(difluoromethyl)ornithine (DFMO), these alpha-methylated polyamines were able to restore cell growth to that observed in the absence of DFMO. In accord with the enzyme data noted above, metabolic studies indicated a slow conversion of 2 to 3, but no metabolism of 4 in these cells. It was concluded from these results that the alpha-methylated polyamines are able to substitute for the natural polyamines spermidine and spermine in critical biochemical processes which involve polyamines for continued cell growth. In accord with the hypothesis, preliminary data indicate that MeSpd and Me2Spm are as effective as spermidine and spermine, respectively, in promoting the conversion of B-DNA to Z-DNA.     

Effect of a new potent H2-receptor antagonist 3[[[2-[(diaminomethylene)amino]-4-thiazolyl]methyl]thio]-N2-sulfamoylpropionamidine (YM-11170) on gastric mucosal histamine-sensitive adenylate cyclase from guinea pig

The effect of 3[[[2-[(diaminomethylene)amino]-4-thiazolyl]methyl]thio]-N²-sulfamoylpro-pionamidine (YM-11170), a new thiazole H₂-receptor antagonist bearing propionamidine at the terminus of a side chain, on histamine-sensitive adenylate cyclase [ATP pyrophosphate-lyase (cyclizing); EC 4.6.1.1] of gastric mucosa from the guinea pig was studied and compared with that of cimetidine. YM-11170 displaced the concentration-stimulation curve of histamine-sensitive adenylate cyclase to the right with a pA₂ of 7.65 (K_i, = 2.25 × 10^?8M). Stimulation of gastric adenylate cyclase by 0.1 mM histamine was competitively inhibited by YM-11170 and cimetidine in a dose-dependent manner, with ic₅₀ values of 5.9 × 10^?7M and 1.4 × 10^?5M respectively. Hippocampal histamine-sensitive adenylate cyclase in the presence of 0.1 mM histamine was also competitively inhibited by YM-11170 with an ic₅₀ of 1.1 × 10^?7 M. YM-11170 did not affect Gpp(NH)p-, NaF-, PGE₂-stimulated or basal activity of the gastric adenylate cyclase. These data, together with other results, indicate that YM-11170 is a highly selective and potent H₂-receptor antagonist which competes with histamine at the receptor site on the histamine-sensitive adenylate cyclase.