Biochemical and pharmacological properties of a new proton pump inhibitor, 2-amino-4,5-dihydropyrido[1,2-a]thiazolo [5,4-g] benzimidazole (YJA20379-5)

This study was designed to determine biochemical and pharmacological properties of a newly synthesized benzimidazole derivative, 2-amino-4,5-dihydropyrido [1,2-a] thiazolo [5,4-g] benzimidazole (YJA20379-5)in vitro andin vivo. In the leaky membrane vesicles of pig gastric mucosa, YJA20379-5 inhibited the K⁺-stimulated H⁺,K⁺-ATPase activity in a concentration- and time-dependent manner, with IC₅₀ values being 43 μM and 31 μM at pH 6.4 and 7.4, respectively. YJA20379-5, given intraduodenally, had a potent inhibitory effect on the gastric acid secretion in pylorus-ligated rats. The ED₅₀ value for acid secretion was 15.4 mg/kg. YJA20379-5, administered orally, also suppressed gastric damages induced by water-immersion stress, indomethacin and ethanol, and duodenal damage induced by mepirizole in rats; the ED₅₀ values were 17.6, 4.7, 3.0 and 18.7 mg/kg, respectively. Furthermore, repeated oral administration of YJA20379-5 accelerated the spontaneous healing of acetic acid-induced gastric ulcers in rats. It is concluded that the antisecretory activity of YJA20379-5 appears to be associated with inhibition of H⁺,K⁺-ATPase, while its antigastric and antiduodenal lesion activities are primarily related to the antisecretory effect.     

Biochemical and pharmacological characteristics of a newly synthesized H+/K+-ATPase inhibitor, YJA20379-8, 3-butyryl-4-[R-1-methylbenzylamino]-8-ethoxy-1,7-naphthyridine, in pigs and rats

We have investigated the effect of the newly synthesized proton-pump inhibitor YJA20379-8, 3-butyryl-4-[R-1-methylbenzylamino]-8-ethoxy-1,7-naphthyridine, on gastric mucosal proton pump (H+/K+-ATPase) activity, gastric acid secretion and gastric lesions in experimental animals. In lyophilized pig gastric microsomes, YJA20379-8 was shown to inhibit H+/K+-ATPase activity; the inhibitory effect was not affected by pH, the IC50 (dose resulting in 50% inhibition) being 28.0 microM and 30.0 microM at pH 6.4 and pH 7.4, respectively. The effect was fully reversed by dilution and subsequent washing of the incubation mixtures of H+/K+-ATPase and YJA20379-8, suggesting the reversible nature of the enzyme inhibition. In pylorus-ligated rats, YJA20379-8 administered by different routes (intraduodenal, subcutaneous, intravenous or oral) resulted in dose-dependent suppression of basal gastric acid secretion. The duration of antisecretory action of 30 mg kg(-1) YJA20379-8 given intraduodenally was very brief (less than 7 h). Pretreatment with YJA20379-8 also dose-dependently prevented gastric lesions induced by absolute ethanol and water-immersion stress in rats. These results suggest that YJA20379-8 might exert its antiulcer activity partly by reversible suppression of acid secretion and partly by protecting the gastric mucosa against ulcerative stimuli.     

Mechanism of antiulcer effect of Neem (Azadirachta indica) leaf extract: effect on H<Superscript>+</Superscript>-K<Superscript>+</Superscript>-ATPase,oxidative damage and apoptosis

The mechanism of the antiulcer effect of Neem leaf aqueous extract to block gastric lesions in rat has been studied with emphasis on acid secretion, oxidative damage and apoptosis. The extract dose-dependently inhibits gastric lesions induced by restraint–cold stress, indomethacin and ethanol. In stress ulcer model, it is more effective than ranitidine but less effective than omeprazole. It also dose-dependently blocks pylorus ligation and mercaptomethylimidazole-induced acid secretion. In the pylorus-ligation model, it is less effective than omeprazole but as effective as ranitidine. It inhibits H⁺-K⁺-ATPase activity in vitro in concentration-dependent manner to inhibit acid secretion. Oxidative membrane damage by hydroxyl radical (^•OH) as measured by lipid peroxidation in stress ulcer is significantly blocked by leaf extract. Stress-induced apoptotic DNA fragmentation is also protected. The extract also prevents ^•OH-mediated mucosal DNA damage in vitro by scavenging the ^•OH. Neem leaf extract, thus, offers antiulcer activity by blocking acid secretion through inhibition of H⁺-K⁺-ATPase and by preventing oxidative damage and apoptosis.     

Inhibitory action of YJA20379, a new proton pump inhibitor onHelicobacter pylori growth and urease

The activities of two types of antiulcer agents against 9 strains ofHelicobacter pylori (H. pylori) were determined by the agar dilution method. The antiulcer agents were YJA20379, a newly synthesized proton pump inhibitor developed by Yung-Jin Pharmaceutical company, and omeprazole. Both compounds were found to have significant activities against this organism. The MIC values of YJA20379 and omeprazole were 11.7 and 31.25 µg/ml, respectively. In addition, the inhibitory potency of both compounds was investigated onH. pylori urease which is believed to be an important colonization and virulence factor in the pathogenesis of gastritis and peptic ulcers. These compounds dose-dependently inhibited urease extracted with distilled water and their IC₅₀ values were 16.4×10^?5 M and 14.3×10^?5 M, respectively. In addition, a pH-dependent study to determine whether inhibitory potency would be activated by acid condition was performed. It was found that unlike omeprazole, YJA20379 was not affected by acid condition. To determine the inhibition pattern and optimal concentration of substrate, kinetics were evaluated at various pH levels (pH 5.0, 7.0, and 8.5). The data show that YJA20379 noncompetitively inhibitedH. pylori urease and K_M/K_i values were 0.96 mM/60 µM (pH 5.0), 0.56 mM/141.5 µM (pH 7.0), and 1.94 mM/34 µM (pH 8.5), respectively. Based on data obtained, it is concluded that YJA20379 is a significant inhibitor ofH. pylori growth and urease and therefore, taking these results into consideration, YJA20379 might be a beneficial therapy for gastritis and peptic ulcers induced byH. pylori.     

Gastric acid secretion in the dog: a mechanism-based pharmacodynamic model for histamine stimulation and irreversible inhibition by omeprazole

A mechanism-based pharmacodynamic model was used to describe the inhibitory effect by omeprazole on gastric acid secretion measured after histamine stimulation in the dog. The model identifies parameters that are related to the physiological system, the histamine stimulation, and the irreversible effect of omeprazole on the H⁺, K⁺-ATPase enzyme. Four different experiments with omeprazole (Exps. 1–4) and two placebo experiments were performed in each of the four Heidenhain pouch dogs used. For placebo and experiments 1–3, saline or omeprazole 0.81 mol/kg was infused during 3 hr with measurements of histamine-stimulated gastric acid secretion in two periods of 3.5–6.5 hr, one period starting just before the omeprazole infusion and a second later period up to 29 hr post infusion. In experiment 4, 0.18 mol/kg of omeprazole was infused for 22.5 min and gastric juice was collected for 5 hr post infusion. The response data was well described by the model. Similar parameter estimates were obtained by three different analysis methods; naïve pooling, two-stage method and nonlinear mixed effects modeling. The elimination rate constant for the H⁺, K⁺-ATPase enzyme, k _out, was estimated to be 0.040 hr^-1, corresponding to a half-life of about 17 hr. This rate constant determines the duration of omeprazole inhibition after long-term exposure. For short-term omeprazole exposure the duration is determined by the rate constant for transfer of enzymes from active to resting state, estimated to be 1.88 hr^-1. The second-order rate constant for histamine stimulation was estimated to be 0.064 hr^-1 per histamine concentration unit and the maximum acid secretion was estimated to be 5.0 mmol H⁺/30 min. The second-order rate constant for the irreversible binding of omeprazole to H⁺, K⁺-ATPase, k _ome, was estimated to be 2.39 L/mol hr. By modeling the histamine-induced baseline response simultaneously with active treatment, predictions of the response are possible not only following different dosing regimens of omeprazole, but also following different degrees of histamine stimulation.     

General pharmacological properties of YJA20379-8, a new H+/K(+)-ATPase inhibitor with anti-ulcer activities

The general pharmacological properties of YJA20379-8 (3-butyryl-4-[(R)-1-methylbenzylamino]-8-ethoxy-1,7-naphthyridine, CAS 187654-40-6), a new H+/K(+)-ATPase inhibitor with anti-ulcer activities, were investigated in mice, rats and guinea pigs. YJA20379-8 at oral doses of 25, 50 and 100 mg/kg did not affect the locomotor activity, hexobarbital hypnosis and motor coordination in mice. The drug did not have analgesic action and anticonvulsant action at the doses of 100 mg/kg p.o. The respiration and blood pressure were not affected at 10 mg/kg i.v. in rats. YJA20379-8 at 2 x 10(-4) g/ml did neither produce any contraction nor relaxation of isolated organs, such as guinea pig ileum, rat fundus, rat uterus and guinea pig vas deferens, and the drug antagonized the contractile responses to several spasmogens, such as acetylcholine, histamine, serotonin, L-phenylephrine, oxytocin and BaCl2. The drug up to 100 mg/kg p.o. did not affect pupil size and the intestinal propulsion of mice. And it did not show an anticarrageenan action at 100 mg/kg. In this general pharmacology study, hypothermic effect in mice, retardation in gastric emptying in rats, decreases in urine excretion in rats at oral doses of 50 and 100 mg/kg of YJA20379-8 and the spasmolytic activity could be found. However, no other effects were exhibited at a high oral dose of 100 mg/kg in animals in this study.     

Cadmium inhibits acid secretion in stimulated frog gastric mucosa

Cadmium, a toxic environmental pollutant, affects the function of different organs such as lungs, liver and kidney. Less is known about its toxic effects on the gastric mucosa. The aim of this study was to investigate the mechanisms by which cadmium impacts on the physiology of gastric mucosa. To this end, intact amphibian mucosae were mounted in Ussing chambers and the rate of acid secretion, short circuit current (I_sc), transepithelial potential (V_t) and resistance (R_t) were recorded in the continuous presence of cadmium. Addition of cadmium (20 µM to 1 mM) on the serosal but not luminal side of the mucosae resulted in inhibition of acid secretion and increase in NPPB-sensitive, chloride-dependent short circuit current. Remarkably, cadmium exerted its effects only on histamine-stimulated tissues. Experiments with TPEN, a cell-permeant chelator for heavy metals, showed that cadmium acts from the intracellular side of the acid secreting cells. Furthermore, cadmium-induced inhibition of acid secretion and increase in I_sc cannot be explained by an action on: 1) H₂ histamine receptor, 2) Ca²⁺ signalling 3) adenylyl cyclase or 4) carbonic anhydrase. Conversely, cadmium was ineffective in the presence of the H⁺/K⁺-ATPase blocker omeprazole suggesting that the two compounds likely act on the same target. Our findings suggest that cadmium affects the functionality of histamine-stimulated gastric mucosa by inhibiting the H⁺/K⁺-ATPase from the intracellular side. These data shed new light on the toxic effect of this dangerous environmental pollutant and may result in new avenues for therapeutic intervention in acute and chronic intoxication.     

Mechanism of action of AZD0865, a K+-competitive inhibitor of gastric H+,K+-ATPase

AZD0865 is a member of a drug class that inhibits gastric H⁺,K⁺-ATPase by K⁺-competitive binding. The objective of these experiments was to characterize the mechanism of action, selectivity and inhibitory potency of AZD0865 in vitro. In porcine ion-leaky vesicles at pH 7.4, AZD0865 concentration-dependently inhibited K⁺-stimulated H⁺,K⁺-ATPase activity (IC₅₀ 1.0 ± 0.2 μM) but was more potent at pH 6.4 (IC₅₀ 0.13 ± 0.01 μM). The IC₅₀ values for a permanent cation analogue, AR-H070091, were 11 ± 1.2 μM at pH 7.4 and 16 ± 1.8 μM at pH 6.4. These results suggest that the protonated form of AZD0865 inhibits H⁺,K⁺-ATPase. In ion-tight vesicles, AZD0865 inhibited H⁺,K⁺-ATPase more potently (IC₅₀ 6.9 ± 0.4 nM) than in ion-leaky vesicles, suggesting a luminal site of action. AZD0865 inhibited acid formation in histamine- or dibutyryl-cAMP-stimulated rabbit gastric glands (IC₅₀ 0.28 ± 0.01 and 0.26 ± 0.003 μM, respectively). In ion-leaky vesicles at pH 7.4, AZD0865 (3 μM) immediately inhibited H⁺,K⁺-ATPase activity by 88 ± 1%. Immediately after a 10-fold dilution H⁺,K⁺-ATPase inhibition was 41%, indicating reversible binding of AZD0865 to gastric H⁺,K⁺-ATPase. In contrast to omeprazole, AZD0865 inhibited H⁺,K⁺-ATPase activity in a K⁺-competitive manner (K_i 46 ± 3 nM). AZD0865 inhibited the process of cation occlusion concentration-dependently (IC₅₀ 1.7 ± 0.06 μM). At 100 μM, AZD0865 reduced porcine renal Na⁺,K⁺-ATPase activity by 9 ± 2%, demonstrating a high selectivity for H⁺,K⁺-ATPase. Thus, AZD0865 potently, K⁺-competitively, and selectively inhibits gastric H⁺,K⁺-ATPase activity and acid formation in vitro, with a fast onset of effect.     

Gastric mucosal blood flow in ethanol-induced mucosal damage in the rat

Gastric mucosal blood flow in ethanol-induced mucosal damage was studied in urethane-anaesthetised rats by reference to ¹⁴C-aminopyrine clearance in the gastric mucosa. Irrigation of the stomach with 30% ethanol in acid saline (100 mM HCl plus 50 mM NaCl) for 40 min broke the gastric mucosal barrier, as indicated by an increased outflow of Na⁺ and K⁺ ions and back-diffusion of H⁺ ions. Gastric mucosal blood flow also increased about 2-fold, decreasing after cessation of ethanol irrigation along with the net ion fluxes. The increase in gastric mucosal blood flow occasioned by 10, 20 and 30% ethanol in acid saline was directly proportional to the net fluxes of H⁺, Na⁺ and K⁺ ions. When the stomach was irrigated with 30% ethanol in a less acid medium (10 mM HCl, 90 mM choline chloride plus 50 mM NaCl) there was still a significant increase in the outflow of Na⁺ and K⁺ ions, but only a slight back-diffusion of H⁺ ions. During this low rate of acid back-diffusion 30% ethanol reduced gastric mucosal blood flow by about 50%. The results suggest that ethanol-induced mucosal damage in the rat is associated with an increase in gastric mucosal blood flow only if combined with back-diffusion of H⁺ ions.     

Bisabolangelone, a gastric H+/K+-ATPase inhibitor: homology modeling and docking study

The homology model of human gastric H⁺/K⁺-ATPase has been produced based on the template provided by pig gastric H⁺/K⁺-ATPase (PDB code: 3IXZ). After molecular mechanics optimization, induced-fit docking simulation between gastric H⁺/K⁺-ATPase and bisabolangelone that has significantly inhibition activity of H⁺/K⁺-ATPase was performed. The results of ligand docking showed that the binding pocket involves the amino acid residues Asp101, Asp102, Tyr105, Leu106, Val296, Phe297, Met299, Ala300, Tyr764, Tyr767, Leu774, Gly777, Cys778, Ile779, Gln889, Tyr893, and Ile952. The hydrogen bonds are formed between bisabolangelone and the amino acid residues Cys778, Gln889, and Tyr893.     

Gastroprotective and antidepressant effects of a new zinc(II)-curcumin complex in rodent models of gastric ulcer and depression induced by stresses

Curcumin, a yellow pigment found in the rhizome of Curcuma loga, has been used to treat a variety of digestive and neuropsychiatric disorders since ancient times in China. Curcumin can chelate various metal ions to form metallocomplexes of curcumin which show greater effects than curcumin alone. This study investigated the antiulcerogenic and antidepressant effects of a Zn(II)-curcumin complex on cold-restraint stress (CRS)-induced gastric ulcers in rats, and on the forced swimming test (FST), tail suspension test (TST) and 5-hydroxy-l-tryptophan (5-HTP)-induced head twitch test in mice. CRS disrupted the rat mucosal barrier and induced gastric ulcers by decreasing the activities of the antioxidant enzymes, and increasing H⁺-K⁺-ATPase activity and malondialdehyde (MDA) level. Pretreatment with Zn(II)-curcumin (12, 24, and 48 mg/kg) dose-dependently reversed these trends, reduced gastric lesions and H⁺-K⁺-ATPase activity, and increased antioxidant activities compared with control groups. Zn(II)-curcumin significantly increased HSP70 mRNA, and attenuated increased iNOS mRNA in the mucosa. Zn(II)-curcumin (17, 34, and 68 mg/kg) also significantly decreased immobility time in the FST and TST, and enhanced 5-HTP-induced head twitches in mice. These results demonstrate that the Zn(II)-curcumin complex showed significant gastroprotective and antidepressant effects compared with curcumin alone via a synergistic effect between curcumin and zinc.     

General pharmacological properties of YJA 20379-1, a novel proton pump inhibitor with antiulcer activities

The general pharmacological properties of YJA 20379-1 (2-amino-4,5-dihydro-8-phenylimidazo[2,1-b]thiazolo[4,5-g]benzo thi azole), a novel proton pump inhibitor with antiulcer activities, were investigated in mice, rats, guinea pig and rabbits. YJA 20379-1 at oral doses of 50, 100 and 200 mg/kg did not affect the general behaviour, hexobarbital hypnosis, motor coordination and body temperature in mice. The drug does not have analgesic and anticonvulsant action at 200 mg/kg p.o. The locomotor activity was not affected at 100 mg/kg p.o., but at 200 mg/kg, the activity was suppressed. YJA 20379-1 (at 2 x 10(-4) g/ml) did neither produce any contraction nor relaxation of isolated organs such as rat fundus, rat uterus, guinea pig ileum and guinea pig vas deferens, and the drug did not antagonize the contractile response to several spasmogens, such as histamine, acetylcholine, serotonin and oxytocin, and the drug up to 200 mg/kg p.o. did not affect pupil size of mice. The intestinal propulsion in mice was not affected up to 200 mg/kg p.o. The gastric emptying in rats was not affected at 100 mg/kg p.o., even if retardation in gastric emptying occurred at 200 mg/kg. YJA 20379-1 did not show anti-inflammatory action nor did it affect urinary excretion up to 200 mg/kg p.o. From these results, it is suggested that YJA 20379-1 at the high dose of 100 mg/kg p.o. may not exert any adverse effects.     

Structure—activity relationships of omeprazole analogues and their mechanism of action

Omeprazole represents a new class of gastric acid secretion inhibitors, the sulphinylbemimidazoles. Per Lindberg and colleagues describe this first example of a clinically useful anti-ulcer agent in this group whose mechanism involves highly specific inhibition of H⁺/K⁺-ATPase, the gastric proton pump. Omeprazole is activated only at low pH, as occurs in the parietal cell and is thus highly selective. The active intermediate formed, a sulphenamide, reacts with a mercapto group of H⁺/K⁺-ATPase to form a disulphide inhibitor complex. The quantity of this sulphenamide on the luminal side of the parietal cell is related to the H⁺/K⁺-ATPase inhibitory effect.     

The long-lasting effect of TU-199, a novel H+, K(+)-ATPase inhibitor, on gastric acid secretion in dogs.

We have used Heidenhain-pouch dogs to investigate the effects of (+/-)-5-methoxy-2-{[(4-methoxy-3,5-dimethylpyrid-2-yl)methyl]sulph inyl}-1H-imidazo[4,5-b]pyridine (TU-199), an imidazopyridine derivative, on gastric acid secretion stimulated by histamine, carbachol and tetragastrin. We have also investigated the duration of the antisecretory effect of TU-199 using a measurement of intragastric pH for 24 h in gastric fistula dogs whose gastric acid secretion was stimulated by histamine. Single oral administration of TU-199 (0.1, 0.2 and 0.4mgkg(-1)) dose-dependently suppressed gastric acid secretion stimulated by histamine infusion. Oral treatment with TU-199 (0.2, 0.4 and 0.8 mg kg(-1)) also dose-dependently inhibited acid secretion induced by carbachol and tetragastrin. The inhibitory effect of TU-199 on stimulated gastric acid secretion was more potent than that of omeprazole, a well-known H+,K(+)-ATPase inhibitor in dogs. Repeated oral treatment with TU-199 at a dose of 0.2 mg kg(-1) once a day for seven days markedly suppressed histamine-stimulated gastric acid secretion in dogs. This inhibitory effect of TU-199 reached a maximum level after three or four doses and was more pronounced than that of omeprazole or lansoprazole. In gastric fistula dogs, the duration of intragastric pH-elevation by administration of TU-199 (0.3 mg kg(-1)) was much longer than that of omeprazole (0.6mgkg(-1)) or lansoprazole (0.9mgkg(-1)). The IC50 values (doses resulting in 50% inhibition) of TU-199, omeprazole and lansoprazole with regard to H+,K(+)-ATPase activity in dog gastric mucosal microsomes were 8.6, 8.8 and 9.9 microM, respectively. These results indicate that TU-199 inhibits gastric acid secretion via suppression of a H+,K(+)-ATPase activity. Our findings also suggest that TU-199 might have potent and long-lasting effects on gastric acid secretion.     

Gastroprotective effect of anti-cancer compound rohitukine: possible role of gastrin antagonism and H<Superscript>+</Superscript> K<Superscript>+</Superscript>-ATPase inhibition

The present study was designed to evaluate the anti-ulcerogenic properties of an alkaloid chromane, rohitukine from Dysoxylum binectariferum. Anti-ulcer potential of rohitukine was assessed in cold restrained, pyloric ligated and ethanol induced ulcers in rats. In addition, rohitukine was tested in vitro for H⁺ K⁺-ATPase inhibitory activity in gastric microsomes. Moreover, we studied the role of rohitukine on the cytosolic concentration of Ca²⁺ in parietal cell-enriched cell suspension in order to ascertain its mechanism of action. Cytoprotective activity was evaluated through PGE₂ level. Rohitukine significantly attenuated the ulcers in cold restraint ulcer (CRU) model in a dose-related manner. Moreover, it significantly lowered the free acidity and pepsin activity in pyloric ligated rats while improved the depleted level of mucin. Furthermore, rohitukine significantly reversed the cold restrained-induced increase in gastrin level. Our in vitro study revealed that rohitukine moderately inhibited the microsomal H⁺ K⁺-ATPase activity with respect to positive control omeprazole. Furthermore, rohitukine potently antagonized the gastrin-elicited increase in cytosolic Ca²⁺ level in parietal cell-enriched suspension. In ethanol-induced gastric lesions in rats, rohitukine significantly inhibited the formation of erosions and increased PGE₂ content showing more potency than reference drug sucralfate. Our results thus suggest that rohitukine possess significant anti-ulcer and anti-gastrinic activity in rats. It is likely that gastro-protective influences of rohitukine are dependent partly on its acid-lowering potential and partly on cytoprotective property. The acid-reducing effect of rohitukine might be attributed to its lowering effect on gastrin production and/or antagonism of gastrin-evoked functional responses of parietal cells. Thus, rohitukine represent a useful agent in the treatment of peptic ulcer disease.     

Proton Pump Inhibitors and Acid-Related Diseases

Proton pump inhibitors (PPIs) are targeted to the gastric acid pump, H⁺,K⁺-adenosine triphosphatase (ATPase). The drugs accumulate in the acid space of the parietal cell and convert to active sulfenamide by an acid-catalyzed reaction. Consequent covalent inhibition of H⁺,K⁺-ATPase blocks the final step of acid secretion, hence the PPIs omeprazole, lansoprazole, and pantoprazole are more effective than histamine₂-receptor antagonists (H₂RAs) in controlling acid secretion. Preclinical short- and long-term clinical surveillance data show these drugs to be well tolerated and safe. The PPIs heal the lesions of gastroesophageal reflux disease and lessen symptoms more effectively and more quickly than the H₂RAs, and are effective' and faster acting for peptic ulcer disease. Helicobacter pylori is causally implicated in the majority of peptic ulcers and in atrophic gastritis. Since PPIs, but not H₂RAs, are synergistic with antibiotics in eradicating H. pylori, their use is appropriate in all acid-related diseases since all patients who are H. pylori positive require eradication as well as healing.     

Sennoside-induced Secretion is Not Caused by Changes in Mucosal Permeability or Na+,K+-ATPase Activity

Abstract— The effect of sennosides (50 mg kg^?1) on the rat colon in-situ was studied 6 h after oral treatment when the laxative effect was maximal. In a second experiment, rhein (4 × 10^?3 m ), an active sennoside metabolite, was administered into the lumen of the colon for 1 h. Both sennosides and rhein reduced net H₂O and Na⁺ absorption or reversed it to net secretion. Paracellular permeability, as measured using erythritol as a small marker molecule, was increased 2- to 3-fold; permeability to a large molecule, PEG 1000, was unchanged. The activity of Na⁺, K⁺-ATPase in the colon mucosa was not affected. There was no damage of the epithelial cells as determined by lactic acid dehydrogenase release. These results indicate that neither inhibition of Na⁺, K⁺-ATPase nor damage of the colon epithelium are involved in the secretory effect of sennosides or rhein. The increased paracellular permeability of small molecules fits into the concept of stimulation of active chloride secretion by sennosides, which is electrochemically and osmotically balanced by an increase in Na⁺ and H₂O flow via the paracellular pathway.     

Nitric oxide,prostaglandin, and sensory neurons in gastric mucosal blood flow response during acid secretion in rats

• 1.1. The mechanism underlying the increase of gastric mucosal blood flow (GMBF) during acid secretion induced by pentagastrin was investigated in anesthetized rats, in relation to nitric oxide (NO), prostaglandin (PG), and sensory neurons.
• 2.2. An intravenous infusion of pentagastrin at 60 μg/kg/h (submaximal dose) produced an increase of acid secretion and GMBF as determined by laser Doppler flowmetry, and the GMBF response was totally attenuated when the acid secretion was inhibited by omeprazole or when the luminal H⁺ was removed by mucosal perfusion with glycine (200 mM).
• 3.3. Prior administration of N^G-nitro-L-arginine methyl ester (L-NAME, 5 mg/kg, IV), a NO synthase inhibitor, significantly mitigated the GMBF response to pentagastrin, without any influence on acid secretion, and this effect was antagonized by coadministration of L-arginine (500 mg/kg IP).
• 4.4. The increase of GMBF during pentagastrin infusion also was significantly mitigated by indomethacin (5 mg/kg, SC) or sensory deafferentation following capsaicin pretreatment, had no effect on the acid secretion, and was totally inhibited by the combined treatments with indomethacin plus L-NAME in addition to sensory deafferentation.
• 5.5. Pentagastrin infusion for 8 hr did not by itself cause any macroscopic damage in the stomach, but additional treatments with L-NAME, and indomethacin plus sensory deafferentation provoked severe lesions in the gastric mucosa.
• 6.6. These results suggest that the increase of GMBF induced by submaximal dose of pentagastrin totally depends on luminal H⁺. This process seems to be mediated by endogenous NO and PGs, as well as capsaicin-sensitive sensory neurons, and to play a pivotal role in maintaining mucosal integrity during acid secretion.

     

Inhibition of gastric K+ ATPase by phenylbutazone and indomethacin.

Inhibition of gastric H⁺ secretion by phenylbutazone and indomethacin was investigated by examining the effects of these agents on a putative H⁺ transport enzyme, a K⁺ stimulated ATPase. unique to gastric mucosa. Phenylbutazone and indomethacin were found to inhibit both the K⁺ ATPase and the K⁺ pNPPase. K_is were 430 μM and 710 μM for the K⁺ ATPase and 330 μM and 670 μM for the K⁺ pNPPase for phenylbutazone and indomethacin respectively. Inhibition was not reversed by Mg²⁺, ATP, pNPP, or KCl and obeyed non-competitive kinetics. Inhibition of the pNPPase suggested that the mechanism of inhibition involved the K⁺ sensitive dephosphorylation of the phosphoenzyme. In the presence of 500 μM phenylbutazone dephosphorylation was significantly less at 3, 5, 7.5, 10 and 15 sec following KCl addition. These studies provide an alternate mechanism for inhibition of gastric H⁺ secretion by phenylbutazone and indomethacin.     

General pharmacological properties of YJA20379-2, a new antiulcer agent

The general pharmacological properties of YJA20379-2 2-dimethylamino-4,5-dihydrothiazolo[4,5:3,4]pyridol[1,2-a]++ +benzoimidazole, a novel proton pump inhibitor with antiulcer activities were investigated in mice, rats, guinea pigs and rabbits. YJA20379-2 at oral doses of 50, 100 and 200 mg/kg did not affect the general behaviour, hexobarbital hypnosis and motor coordination in mice. The drug did not have analgesic or anticonvulsant action at 200 mg/kg. Locomotor activity and body temperature were not influenced at 100 mg/kg. At a concentration up to 2 x 10(-4) g/ml, YJA20379-2 did not produce any contraction or relaxation of isolated preparations, such as the rat fundus, the guinea pig ileum and the rat uterus, and did not antagonize the contractile response to several spasmogens, such as histamine, acetylcholine, serotonin and oxytocin. At dosages up to 200mg/kg p.o. YJA20379-2 did not affect the pupil size of mice. Intestinal propulsion of mice was not affected up to 200 mg/kg p.o. and the drug did not affect urinary excretion at 100 mg/kg p.o. These results indicate that at dosages up to 100 mg/kg p.o. YJA20379 was found not to affect this pharmacological profile. However, at 200 mg/kg the drug lowered body temperature and showed decreases in locomotor activity and urine volume.