In vitro activity of sertaconazole, fluconazole, ketoconazole, fenticonazole, clotrimazole and itraconazole against pathogenic vaginal yeast isolates.

The in vitro activity of sertaconazole was compared with those of the most commonly used vaginal antimycotic agents--fluconazole, ketoconazole, fenticonazole, clotrimazole and itraconazole--against 94 strains of clinical isolates of Candida spp. using a macrodilution method in Casitone agar medium. The sertaconazole concentration (microgram/ml), at which 90% of the strains were inhibited, was 0.06 for C. albicans, 0.25 for C. glabrata and C. parapsilosis, 1 for C. krusei and 2 for C. tropicalis. These values show that sertaconazole is one of the most active products against yeasts causing vulvovaginal candidiasis, its activity against C. glabrata being particularly relevant.

Cytidine (5')diphosphocholine modulates dopamine K(+)-evoked release in striatum measured by microdialysis.

Experiments were performed to determine whether exogenous cytidine (5')diphosphocholine (CDP-choline) could modify the release of dopamine (DA) in the striatum. Rats were divided into two groups, receiving either a standard diet (UAR 004) or the same diet supplemented with CDP-choline (250 mg/kg day) for 28 days. On the last day the dialysis probes were inserted in the striatum, and DA, homovanillic acid (HVA), and 3,4-dihydroxyphenylacetic acid (DOPAC) efflux were measured in the dialysis stream basally and during K+ depolarization (80 mM K+). Basal DA, HVA, and DOPAC did not show any difference between treated and untreated groups. Depolarization with K+ increased DA levels by up to 3,000% in the control group and by up to 4,770% in the CDP-choline-treated group (p < 0.05), and reduced extracellular HVA and DOPAC concentration by up to 45 and 35%, respectively, both in the untreated and CDP-choline-treated groups. These results show that long-term treatment with CDP-choline increases the K+ induced release DN and suggest, in accordance with previous research, that by providing exogenous choline and cytidine, CDP-choline modulates dopaminergic transmission.

7-[3-(1-piperidinyl)propoxy]chromenones as potential atypical antipsychotics. 2. Pharmacological profile of 7-[3-[4-(6-fluoro-1, 2-benzisoxazol-3-yl)-piperidin-1-yl]propoxy]-3-(hydroxymeth yl)chromen -4-one (abaperidone, FI-8602).

A series of novel 7-[3-(1-piperidinyl)propoxy]chromenones was synthesized and tested as potential antipsychotics in several in vitro and in vivo assays. The compounds possessed good affinity for D2 receptors, together with a greater affinity for 5-HT2 receptors, a profile which has been proposed as a model for atypical antipsychotics. Several agents also displayed a high potency in the climbing mice assay on oral administration, suggesting a potent antipsychotic effect as compared to reference standards. Compound 23 was selected for further pharmacological evaluation. Induction of catalepsy and inhibition of stereotypies weaker than standards, along with a lower increase in serum prolactin levels, were indicative of a potential atypical profile for this compound. From these results, 7-[3-[4-(6-fluoro-1, 2-benzisoxazol-3-yl)piperidin-1-yl]propoxy]-3-(hydroxymethyl )chromen- 4-one (23, abaperidone) has been proposed for clinical evaluation in humans as a potential atypical antipsychotic.

Histamine H2-receptor antagonist action of ebrotidine. Effects on gastric acid secretion, gastrin levels and NSAID-induced gastrotoxicity in the rat.

The antagonism of histamine H2-receptors by ebrotidine (N-[(E)-[[2-[[[2-[(diaminomethylene)amino]-4-thiazolyl]methyl]thio]ethyl ] amino]methylene]-4-bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542) was assessed on isolated guinea-pig right atrium. The dose-response curves obtained by histamine on the positive chronotropic effect in guinea-pig atrium were displaced to the right in parallel depending on the concentration of ebrotidine and ranitidine without change in the maximum response with pA2 values of 7.12 and 7.26, respectively. The slope of the regression line of log (DR-1) against log ebrotidine concentration was not significantly different from unity: 0.96 (95% confidence limits: 0.89-1.03). These results indicate that ebrotidine is a competitive H2-receptor antagonist. Following intravenous administration to rats, ebrotidine inhibited histamine- and pentagastrin-stimulated acid secretion in a dose-dependent manner, ED50 being 0.21 and 0.44 mg/kg, respectively. After oral administration to fasting rats 3 h before their sacrifice, ebrotidine decreased the total acid contents of the stomach in a dose-dependent manner, ED50 being 7.5 mg/kg. After a single dose of 100 mg/kg in fasting rats, ebrotidine increased significantly serum gastrin levels within 2 and 5 h after administration, but 8 h after administration serum gastrin levels returned to normal values. In contrast, ranitidine at a single oral dose of 100 mg/kg increased serum gastrin levels more markedly within 2 and 5 h after administration, while after 8 h, this increase still persisted although without significant differences with respect to control, and after 24 h levels returned to normal values. Both ebrotidine and ranitidine were administered orally at a dose of 100 mg/kg for 26 days showing significant increments in plasma gastrin levels 5 h after administration. Such increments were not so marked after ebrotidine and normal values were attained at 24 h after administration. The results obtained after repeated oral administration for 15 days of ebrotidine and ranitidine at the doses of 15 and 50 mg/kg demonstrated that ebrotidine did not increase significantly serum gastrin levels with respect to control 2 h after administration, and no dose-related effect was observed. In contrast, ranitidine increased serum gastrin levels significantly and in a dose-dependent manner with respect to control group. ED50 values of ebrotidine obtained in the experiments on the prevention of NSAID-induced gastrotoxicity in the rat were 12.2, 12.5, 11.5 and 9.8 mg/kg against diclofenac, ketoprofen, indometacin and naproxen, respectively. ED50 values of ranitidine were of the same order: 20.6, 13.9, > 50 and 15.1 mg/kg.

Action of ebrotidine, ranitidine and cimetidine on the specific binding to histamine H1- and H2-receptors.

Ebrotidine (N-[(E)-[[2-[[[2-[(diaminomethylene)-4-thiazolyl]methyl]thio] ethyl]amino]methylene]-4-bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542), a selective H2-receptor antagonist, has proved to competitively inhibit the positive chronotropism induced by histamine in isolated guinea pig atrium. The affinity of ebrotidine to histamine H1- and H2-receptors through the displacement of 3H-pyrilamine and 3H-thiotidine binding to guinea pig cerebellum and brain cortex membranes was investigated. Ebrotidine displaced 3H-thiotidine specific binding to histamine H2-receptors (Ki: 127.5 nmol/l), showing a higher affinity (p < 0.05) than ranitidine (Ki: 190.0 nmol/l) and cimetidine (Ki: 246.1 nmol/l). None of the three substances displaced 3H-pyrilamine binding to H1-receptors (Ki: > 5000 nmol/l). The results showed that ebrotidine is a drug with a high affinity for H2 receptors, higher than cimetidine and ranitidine.

Antiinflammatory activity of topically applied sertaconazole nitrate.

The antiinflammatory activity of topically applied sertaconazole nitrate was investigated. The dermal administration of 2% sertaconazole to rats, in which edema was induced by application of croton oil in the external ear, induced a 39.8% reduction of edema. The results show that sertaconazole applied by topical route at the concentration at which it is used in clinic, has antiinflammatory action in this experimental model.

Evidence that 5'-cytidinediphosphocholine can affect brain phospholipid composition by increasing choline and cytidine plasma levels.

We examined the effects of orally administered 5'-cytidinediphosphocholine (CDP-choline) on arterial plasma choline and cytidine levels and on brain phospholipid composition in rats. Animals receiving a single oral dose of 100, 250, or 500 mg/kg showed peak plasma choline levels 6-8 h after drug administration (from 12 +/- 1 to 17 +/- 2, 19 +/- 2, and 24 +/- 2 microM, respectively). The area under the plasma choline curve at > 14 microM, i.e., at a concentration that induces a net influx of choline into the brain, was significantly correlated with CDP-choline dose. In rats receiving 500 mg/kg this area was 2.3 times that of animals consuming 250 mg/kg, which in turn was 1.8 times that of rats receiving 100 mg/kg. Plasma cytidine concentrations increased 5.4, 6.5, and 15.1 times baseline levels, respectively, 8 h after each of the three doses. When the oral CDP-choline treatment was prolonged for 42 and 90 days, brain phosphatidylcholine concentrations increased significantly (by 22-25%; p < 0.05) in rats consuming 500 mg/kg/day. Brain phosphatidylethanolamine and phosphatidylserine concentrations also increased significantly under some experimental conditions; levels of other phospholipids were unchanged.

Oral cytidine 5'-diphosphate choline administration to rats increases brain phospholipid levels.

Exogenous cytidine 5'diphosphocholine (CDP-choline) is completely metabolized to circulating cytidine and choline. Both compounds enter the brain and can be used in phosphatidylcholine (PC) synthesis via the Kennedy (CDP-choline) cycle. We administered oral CDP-choline to 12 month-old rats (500 mg/kg/day) for 21, 42, or 90 days to determine whether this treatment would alter brain levels of PC and the other structural phospholipids, phosphatidylserine (PS) and phosphatidylethanolamine (PE). After 42 days, brain PC levels increased significantly (p < 0.01) by 23.3%; after 90 days PC increased by 30% (p < 0.01), PS by 37.2% (p < 0.01), and PE by 13% (not significant). The ratios of each of the phospholipids to total membrane phospholipids were unchanged. These data demonstrate that repeated oral CDP-choline administration can increase the amounts of phospholipids in brain membranes, thus providing a rationale for using this compound in brain diseases that damage neurons.

Effects of orally administered cytidine 5'-diphosphate choline on brain phospholipid content.

Cytidine, as cytidine 5'-diphosphate choline (CDP-choline), is important for the synthesis of phosphatidylcholine in cell membranes. To investigate whether exogenous CDP-choline could affect brain phospholipid composition, we supplemented the diet of mice with this drug (500 mg/kg/day) for 27 months in 3-month-old mice and for 90, 42, and 3 days in 12-month-old mice, and measured their levels of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), and the content of phosphatidylinositol plus phosphatidic acid in the cerebral cortex. After 27 months of treatment, PC and PE increased significantly by 19% (P < 0.05) and by 20% (P < 0.01), respectively. PS levels increased by 18% (not statistically significant). Similar elevations in PC and PE levels were obtained when older mice were treated for only 3 months (P < 0.05). No changes were observed with shorter treatment periods. These results suggest that chronic administration of CDP-choline can have effects on brain phospholipid composition that may underlie its reported utility in various neurologic disorders.

Inhibition of ergosterol synthesis by sertaconazole in Candida albicans.

7-Chloro-3-[1-(2,4-dichlorophenyl)-2-(1H-imidazol-1-yl) ethoxy-methyl] benzo [b] thiophene (sertaconazole, FI-7045, CAS 99592-32-2), a new topical antifungal agent, has shown activity against a broad range of clinically relevant yeasts and fungi. In the present study, the molecular basis of the pharmacological action of sertaconazole was examined by investigating the inhibition of ergosterol synthesis in cultures of Candida albicans, ATCC E-10.231, at 6 x 10(5) cells/ml grown aerobically at 37 degrees C, using various concentrations of sertaconazole. Sertaconazole inhibited the ergosterol synthesis with IC50 = 115 nmol/l. The results show that one of the mechanisms of action of sertaconazole consists in the inhibition of ergosterol synthesis.

Direct membrane-damaging effect of sertaconazole on Candida albicans as a mechanism of its fungicidal activity.

The direct action of 7-chloro-3-[1-(2,4-dichlorophenyl)-2-(1H-imidazol-1-yl)ethoxy- methyl]benzo[b]thiophene (sertaconazole, FI-7045, CAS 99592-32-2) on the membrane integrity of C. albicans is studied by quantifying the leakage of intracellular adenosine triphosphate (ATP) into the medium as an index of the changes in membrane permeability and integrity and cell viability of the culture used. Sertaconazole caused a dose-dependent decrease in intracellular ATP after only 10-min exposure and a concomitant significant increase in extracellular ATP. This behaviour is characteristic of antifungals which are fungicidal as a result of a direct membrane damage. Thus sertaconazole, in addition to the mechanism of action responsible for its fungistatic activity (inhibition of ergosterol synthesis), has a second mechanism of action providing a significant fungicidal activity due to direct cell membrane damage.

Pharmacokinetic evaluation of labelled sertaconazole after dermal application.

14C-labelled 7-chloro-3-[1-(2,4-dichlorophenyl)-2-(1H-imidazol-1-yl)ethoxy-methyl] benzo[b]thiophene (sertaconazole, FI-7045, CAS 99592-32-2) was administered systemically (i.v. route) and topically (dermal route) in different formulations at doses of 10 mg/kg to hairless Sprague-Dawley rats. Determination of plasma and liver levels of radioactivity demonstrated that the dermal application of sertaconazole induced very low systemic absorption with all the formulations used: 1.47% for sertaconazole cream, 1.97% for sertaconazole solution, 0.665% for sertaconazole powder and 0.885% for sertaconazole gel. Calculations were made from the areas under plasma levels curves of each topical administration in comparison with intravenous administration. Significant radioactivity levels were found in the liver, which was consistent with a high hepatic metabolization and faecal excretion of the substance. However, the liver levels after topical application were virtually insignificant in comparison with the levels in the same organ obtained after intravenous administration.

Age-related changes in memory and their pharmacologic modulation.

A passive avoidance procedure was used to measure the ability to learn and remember to avoid a noxious stimulus in 13-month-old mice and to test the effects that treatment with cytidine(5')-diphosphocholine (CDP-choline) had on these processes. Half of the mice received 500 mg/kg per day CDP-choline orally for 4 months, and the two groups were compared with a third group of younger animals (4-month-old mice). The older mice showed marked impairment in the execution of these tests; however, those treated with CDP-choline had significant improvement in their performance 24 hours after learning the task.

Metabolism of cytidine (5?)-diphosphocholine (cdp-choline) following oral and intravenous administration to the human and the rat.

We examined the effects of cytidine (5?)-diphosphocholine (CDP-choline) on plasma levels of cytidine, choline, and unchanged CDP-choline among normal volunteers receiving the substance orally or intravenously, and rats receiving it intravenously. Two hours after a single oral dose (2g), plasma choline levels were increased by 48% and plasma cytidine by 136%. Among subjects receiving three doses (2g each) at two-hour intervals, plasma choline peaked (30% over baseline) 4 h after the initial CDP-choline dose, while plasma cytidine levels continued to increase for at lest 6 h, at which time they were five times basal levels (P < 0.01). Intravenously-administered CDP-choline was rapidly hydrolysed, in both the human and the rat. In humans given the CDP-choline by infusion over 30 min, plasma CDP-choline fell to undetectable levels almost immediately after the end of the infusion period; plasma choline and cytidine peaked at that time, but their concentrations remained significantly elevated for at least 6 h. In rats given a bolus injection of CDP-choline, five minutes earlier, the unchanged compound was also undetectable in plasma, while plasma cytidine levels increased markedly and remained elevated for at least 60 min. These observations show that CDP-choline is converted to at least two major circulating metabolites, choline and cytidine. Since both of these compounds are used in the biosynthesis of phosphatidylcholine, both may be involved in the long-term effects of the CDP-choline.

Effect of cytidine(5')diphosphocholine (CDP-choline) on the total urinary excretion of 3-methoxy-4-hydroxyphenylglycol (MHPG) by rats and humans.

We examined the effects of orally administered cytidine(5')-diphosphocholine (CDP-choline) on the total levels of 3-methoxy-4-hydroxyphenylglycol (MHPG) in human and rat urine. Four subjects who had been on a low-choline diet (less than 1 gm/day) for 24 hours received three doses of CDP-choline (2 gm each) at 8 a.m., 10 a.m., and noon; urines were collected at two-hour intervals after each dose. Rats received water for three days; then CDP-choline (100 mg/kg) or equimolar doses of choline for five days; then water again for three more days. Twenty-four hour urine samples were collected on each day of the study. The levels of MHPG in human urine increased by 45-68% when subjects were receiving CDP-choline (p less than 0.01). CDP-choline, but not choline, also elevated urinary MHPG significantly in rats (p less than 0.01). These data suggest that CDP-choline enhances norepinephrine release, and that this action may be mediated by more than just its choline content.

Cytidine(5')diphosphocholine enhances the ability of haloperidol to increase dopamine metabolites in the striatum of the rat and to diminish stereotyped behavior induced by apomorphine.

Experiments were performed to determine whether exogenous cytidine(5')diphosphocholine (CDP-choline) could modify release of dopamine in the striatum and behavior dependent on dopamine, perhaps by providing supplemental choline for synthesis of acetylcholine. Rats received water (control) or CDP-choline orally (100 mg/kg per day, for 5 days), either alone or before injection with haloperidol (1 mg/kg, i.p.), apomorphine (0.15 mg/kg, s.c.), or both. Stereotyped behavior was measured during the hour after administration of apomorphine; levels of the dopamine metabolites, homovanillic acid (HVA) and 3,4-dihydroxyphenylacetic acid (DOPAC) in the striatum were assessed at the end of this period (2 hr after administration of haloperidol). In rats receiving the CDP-choline, the stereotyped behavior observed after injection of apomorphine alone (P less than 0.01), or after haloperidol plus apomorphine (P less than 0.01), was attenuated. The pretreatment with CDP-choline also significantly increased levels of HVA (by 24%) and DOPAC (by 23%) in the striatum over appropriate controls in animals receiving haloperidol, or by 29 and 59% (averaging data for all time points), respectively, in animals receiving haloperidol plus apomorphine. One mechanism by which CDP-choline may affect behavior involves contributing choline to enhance synthesis of acetylcholine.

Cerebral subcellular distribution of CDP-choline and/or its metabolites after oral administration of methyl-14C CDP-choline.

Distribution of radioactivity in subcellular components of a cerebral extract after oral administration of methyl-14C labelled cytidine diphosphate choline (CDP-choline, citicoline, Somazina) was analyzed. Animals were decapitated 24 h after the administration and the brain immediately removed and gently homogenized in a buffer solution. The homogenate was fractionated by differential centrifugation and the protein concentration and the radioactivity were measured in each precipitate as well as in the final supernatant. Radioactivity was found to be associated mainly with membranous microsomal precipitate and with cytoplasmic membrane precipitate. Alternatively, materials sedimenting between 500 X g and 20,000 X g were fractionated in a density gradient centrifugation. Fractions were screened for enzymatic activities as indicators of cytoplasmic membrane, mitochondria, lysosomes, microsomes and synaptosomes. Radioactivity was found to be associated with cytoplasmic and microsomal membrane fractions. Identification of some fractions as enriched in synaptosomes also accumulating radioactivity, as well as the possible incorporation of the radioactivity to choline and phosphorylcholine cerebral pools are discussed.

Dissimilar effects in acute toxicity studies of CDP-choline and choline.

The LD50 and LD0 of choline HCl and cytidine diphosphate choline (CDP-choline, citicoline, Somazina) by oral and intravenous route at equivalent doses of choline for both compounds were determined. The qualitative and quantitative analysis of the results obtained lead to the conclusion that choline shows a remarkable cholinergic action, while CDP-choline both orally and intravenously does not cause any cholinergic intoxication in the treated groups, like those recorded for choline administration. It should be emphasized that when the quantity of choline contained in CDP-choline administered was identical to that of choline with which the animal group was treated showing signs of cholinergic crisis, the animals corresponding to CDP-choline did not show any of these symptoms. Objective evaluation showed that CDP-choline by oral and intravenous route does not behave like a choline groups donator, at least not with an intensity which would make us believe that its only action was due to the increased levels of choline; this proves that CDP-choline administration yields clearly differentiated metabolic consequences from those yielded by choline administration.

Effect of oral CDP-choline on acrylamide-induced lesion.

Administration of low doses of cytidine diphosphate choline (CDP-choline, citicoline, Somazina) (50 mg/kg) to mice and rats has proved to be effective in preventing acrylamide-induced neurological syndrome. Likewise, the simultaneous dosing of both drugs, which causes an evident loss of weight in the animals, has demonstrated that an activation on the dopaminergic system is produced.

Bioavailability of methyl-14C CDP-choline by oral route.

In this work, the bioavailability of the 14C-methyl-labelled cytidine diphosphate choline (CDP-Choline, citicoline, Somazina), has been studied by oral route, leading to the result that it is slowly and completely absorbed, with a very low urinary and fecal elimination, thus producing maintained blood levels. The bioavailability related to oral route compared with intravenous route is practically the same.