Belladonna alkaloids-induced behavioral changes and amnesia on open-field and step-through in 18-,28-, and38-day-old mice

目的：比较阿托品（Ａｔｒ），东莨菪碱（Ｓｃｏ），樟柳碱（ＡＴ３）和山莨菪碱（Ａｎｉ）对小鼠行为及记忆损伤作用．方法：行为和记忆实验用开阔和回避反应法．脑Ｍ受体用［３Ｈ］ＱＮＢ测定．结果：Ａｔｒ，Ｓｃｏ和ＡＴ３增加小鼠走动行为２６％－４２％，降低站立，修饰，排便行为５０％－１００％，并抑制开阔记忆．４个药物均能妨碍回避反应．小鼠１８日龄额叶皮层和海马［３Ｈ］ＱＮＢ结合位点数少于３８日龄７％－２３％．结论：１）莨菪类生物碱对小鼠行为和记忆障碍的作用随其日龄增加而减弱．２）Ｓｃｏ对幼年小鼠的行为及记忆障碍作用的最小有效量分别是Ａｔｒ，ＡＴ３和Ａｎｉ的１／１０，１／１００和１／１０００．     

Kinetics and metabolism of (+)-, (−)- and (±)-bufuralol

Summary Single oral doses of (+)-, (–)- and (±)-bufuralol were administered to a healthy volunteer to compare the disposition and metabolism of the individual isomers and the racemate. Plasma levels and area under plasma curve (AUC) of the active isomer, (–)-bufuralol, were higher than those of the (+)-isomer; plasma clearance was correspondingly lower. Intermediate values were found for the racemate. The elimination half-life of (–)-bufuralol was shorter than that of (+)-bufuralol, but similar to the racemate. Both isomers were cleared almost entirely by metabolism. The main metabolic pathway for (–)-bufuralol was aromatic hydroxylation, whereas the principal route for (+)-bufuralol was conjugation. Phenol metabolites in the systemic circulation were present mainly as conjugates. Both isomers also underwent aliphatic hydroxylation. This pathway was more favoured by the (+)-isomer, although plasma levels and AUC of the principal product, 2-hydroxy-bufuralol, were almost identical for the two forms. Major differences in metabolic fate thus had relatively little effect on the disposition of pharmacologically active metabolites.     

rac- and meso-Cyclohexanoids: Their α-, β-glycosidases,antibacterial, antifungal activities,and molecular docking studies

An efficient and versatile synthesis method has been postulated for hydroxymethylated rac - and meso-cyclohexanoid derivatives. The synthesis of these stereoisomers was achieved easily with traditional methods using hexahydroisobenzofuran 6 , prepared from commercially available cis-hydrophthalic anhydride. The study, involving diastereoselective epoxidation and cis-hydroxylation, was conducted to obtain epoxy-, cis-, and trans-diol-furans 7, 8 , and 9 . After sulfamic acid-catalyzed ring-opening reaction of the epoxide and furan rings, rac- and meso-tetraacetates 14, 15 , and 16 were afforded. Hydrolysis of acetate groups with ammonia in absolute methanol yielded the desired tetrols rac -17 , meso -18 , and meso -19 . All structures, after purification by chromatographic methods and elucidation by spectral techniques, were screened against α- and β-glucosidases. Compounds 7, 8, 10, 17, 18 , and 19 were also evaluated for their antibacterial and antifungal activity against some selected synthesized compounds with varying degrees of inhibitory effects on the growth of different pathogenic microorganisms by the well-diffusion method. In addition, Saccharomyces cerevisiae α-glucosidase molecular modeling studies were performed for all rac- and meso-compounds 7, 8, 10, 17, 18 , and 19 .     

Study of the inclusion complexes formed between cetirizine and α-, β-, and γ-cyclodextrin and evaluation on their taste-masking properties

Complexation properties of cetirizine dihydrochloride (cetirizine) with α-, β-, and γ-cyclodextrin (CD) were investigated by ultra violet (UV) and nuclear magnetic resonance (NMR) spectroscopies and isothermal titration calorimetry (ITC). The use of the continuous variation method, applied on UV and NMR data, demonstrated 1:1 complex stoichiometry for cetirizine-α-CD, cetirizine-β-CD, and cetirizine-γ-CD, respectively. NMR two-dimensional Rotational nuclear Overhauser Effect SpectroscopY experiments revealed that for α- and β-CD, the complexation takes place by including either the phenyl or chlorophenyl ring of the cetirizine into the CD cavity, whereas in the case of γ-CD, both rings can be included simultaneously. Association constants (K(a)) determined by UV spectroscopy demonstrated that cetirizine forms more stable complex with β-CD (K(a) = 5641 ± 358 M(-1)) than α-CD (K(a) = 1434 ± 60 M(-1)). No information could be extracted from the UV spectroscopic analysis of cetirizine-γ-CD solutions. ITC results for association constant determination were in compliance with UV results and confirmed that the highest association constant was found for the cetirizine-β-CD complex (2540 ± 122 M(-1)). The association constants from ITC measurements for cetirizine-γ-CD and cetirizine-α-CD complexes were found to be 1200 ± 50 and 800 ± 22 M(-1) , respectively. Taste-masking studies revealed that β-CD is the only native CD recommendable for oral pharmaceutical formulations.     

Synthesis and antifungal activity of sulfides,sulfoxides, and sulfones based on (1S)-(-)-β-pinene

Attachment of 2-mercaptoethanol and thioglycolic acid methyl ester to the double bond of (1S)-(-)-β-pinene yielded pinane sulfides with the cis configuration. Oxidation of sulfides with m-chloroperbenzoic acid yielded the corresponding sulfoxides and sulfones. The resulting compounds were screened for antimycotic activity and the dynamics of changes in antifungal properties in sulfides-sulfoxide-sulfone series were studied.     

Synthesis, anti-inflammatory, and cytotoxicity evaluation of 9,10-dihydroanthracene-9,10-α,β-succinimide and bis-succinimide derivatives

9,10-Dihydroanthracene-9,10-α,β-succinimide derivatives 4a–e and bis-succinimide derivatives 6a–e have been synthesized by grinding 9,10-dihydroanthracene-9,10-α,β-succinic anhydride 2 with various mono 3a–e and diamines 5a–e in quantitative yields. All the target compounds were fully characterized by spectrometric and spectroscopic means. Compounds 4a–e, 6a–e and recently reported compounds 4f–p were screened for anti-inflammatory and for cytotoxicity against five human cancer cell lines: T47D, NCI H-522, HCT-15, PA1, and HepG-2. Compounds 4e, 4i, 4j, and 4p exhibited good anti-inflammatory activity and compounds with interesting cytotoxic profile were 4c, 6e (T47D); 4e, 4o (NCI H-522); 4n (HCT-15); 4e, 4h, 4o (PA1); and 4a, 4e, 4f, 4i, 4o (HepG-2).     

Differences in presynaptic α-blockade,noradrenaline uptake inhibition,and potential antidepressant activity between (+)- and (-)mianserin

The effect of racemic mianserin on K⁺-evoked tritium release from rat brain cortex slices previously incubated with ³H-l-noradrenaline was studied. Racemic mianserin (10^-9-10^-5 M) increased stimulation-induced release dose-dependently. As methysergide, metiamide, and cyproheptadine failed to do so, it was concluded that this effect was probably not caused by the antihistamine or antiserotonin activity of racemic mianserin, but due to its -adrenolytic effect. Evaluation of the effects of the enantiomers (+)(S)mianserin and (-)(R)mianserin showed that the -adrenolytic effect resided in the (+)isomer, whereas the (-)isomer was inactive at a concentration of 10^-6 M. Inhibition of noradrenaline into rat hypothalamic synaptosomes also showed stereospecificity in that (+)mianserin was about 300-times more active than (-)mianserin. Inhibition of rat muricidal behavior, a test for potential antidepressant activity, showed a similar dissociation in the effects of the two enantiomers, in that (+)mianserin was active, whereas (-)mianserin was not.     

Effects of Kai - Xin - San, a traditional Chinese medicine, and its four herbal components on learning and memory

Senile dementia disease characterized by memory dysfunction and Alzheimer disease , are becoming more frequent in the aged populations . Modem medicine does not offer medical treatments to the amnesia and dementia. However , in the traditional Chinese medicines, several crude drugs created already thousand years ago , were thought to benefit the brain functions and to improve memory abilities . Kai - Xin -San, has been used since the Tang dynasty , and it has been applied in numerous compositions targeting senile dementia. Even in Japan, Kai - Xin - San has been used since the Heian dynasty . Kai- Xin- San contains ginseng （ panaxginseng C. A. Meyer） , polygala （ Polygala tenaifolia Wildenon） , acorus （ Acorus graminus Soland） , and hoelen （ Poria cocos Wolf） , at a ratio of 1： 1： 25：50 （ dry weight） . The traditional medicine suggested that Kai - Xin - San and four herbal components on memory dysfunctions using several behavioral animal models, as well as electrophysiological models of memory formation （ short - and long term potentiation） and hippocampal neuronal cell culture. Kai - Xin - San has protective effects against ischemia, ameliorated impairment of memory acquisitions induced by alcohol , enhanced recovery of memory functions of amygdale - lesioned mice , improved aging process in senile dementic animal model , and facilitated the hippocampul LTP - induction. Taken together , the resuits suggest that Kai - Xin - San directly -affected the hippocampal synaptic transmission,     

Disposition and metabolism of amosulalol hydrochloride,a new combined α- and β-adrenoceptor blocking agent,in rats,dogs and monkeys

1. The disposition and metabolism of amosulalol hydrochloride, a combined α- and β-adrenoceptor blocking agent, were studied in rats, dogs and monkeys.

2. After oral administration of [¹⁴C]amosulalol hydrochloride, the plasma concentration of radioactivity reached a maximum at 05 to 1 h in all species and declined with half-lives of about 2 h in both rats and monkeys, and of about 4 h in dogs. The ratios of unchanged drug to total radioactivity in the rat and dog plasma were 8 and 43% at 05 h after administration, respectively. The radioactivity in the rat tissues was high in the liver, kidney, blood and pancreas after oral administration.

3. Following oral dosage, the urinary excretion of radioactivity was 26-34% of the dose in rats, 45% in dogs and 46% in monkeys in 48 h. The biliary excretion after oral dosage amounted to 66% and 41% in rats and dogs, respectively.

4. Six metabolites were isolated and identified from the urine of rats and dogs. They were derived from one or two of the following pathways: I, hydroxylation of the 2-methyl group of the methylbenzenesulphonamide ring; II, demethylation of the o-methoxy group of the methoxyphenoxy ring; III, hydroxylation at the 4 or 5 position of the methoxy-phenoxy ring; IV, oxidative cleavage of the C—N bond yielding o-methoxyphenoxy acetic acid. Moreover, some metabolites were metabolized to glucuronide or sulphate.     

Inclusion complexation of glisentide with α-, β- and γ-cyclodextrins

Complexation of glisentide with α-, β- and γ-cyclodextrin (CD) has been investigated in aqueous solution and in the solid state. Complex formation in solution has been analysed using solubility diagrams and NMR spectroscopy and the interaction in solid state has been studied by X-ray diffractometry, DSC and IR spectroscopy. The thermodynamic parameters, ΔH°, ΔS° and ΔG°, of complexation with β- and γ-CD have been calculated from the temperature dependence of the stability constant. The process has been found to be exothermic and ΔS° is slightly unfavourable. In addition, it has been found that the ionization state of glisentide plays an important role in complexation and the fact that the extent of complexation is greater with β- than with γ-CD has revealed the importance of the cavity size to get an adequate fitting between host and guest molecules. The inclusion of the ortho-substituted aromatic ring of glisentide has been evidenced by NMR spectroscopy. Finally, complexes have been prepared by coprecipitation and kneading methods and it has been found that the former is more suitable to achieve solid-state complexation.     

Behavioural effects of selective μ-, κ-, and δ-opioid agonists in neonatal rats

The behavioural effects of selective -, - and -opioid agonists in 5-, 10- and 20-day-old rats were investigated by observational analysis. The predominant response to -agonists was behavioural depression. High doses (10 mg/kg IP) of morphine and DAGO (d-Ala², NMe-Phe⁴, Glyol⁵-enkephalin) produced overt sedation in all the age groups and also induced catalepsy which was particularly apparent in the 5- and 10-day-old animals. These compounds did not produce any signs of behavioural activation in the neonatal rats. In contrast, rat pups treated with the -agonists U50,488H and PD 117,302 (1,10 mg/kg IP) exhibited marked hyperactivity with increases in wall-climbing and locomotion. Sedative effects of the highest dose of the -agonists began to emerge, however, as the animals grew older, resulting in significant decreases in behaviours such as gnawing and grooming at 20 days of age. The -agonist (+)-tifluadom (0.1–10 mg/kg), but not its corresponding (-)-isomer, produced an increase in activity in 5-day-old rats, thus extending the observations made with U50,488H and PD 117,302 and establishing the stereoselective nature of the response. The involvement of -receptors in opioid-induced hyperactivity was further substantiated by using a variety of opioid antagonists. In this context, the increase in activity induced by U50,488H (10 mg/kg) in 5-day-old neonates was attenuated by naltrexone (1 mg/kg IP) but not by larger doses (10 mg/kg) of either M8008 (which has low affinity for -receptors) or the selective -receptor antagonist ICI 174,864. Finally, DPDPE (d-Pen², d-Pen⁵-enkephalin) which acts selectively at -opioid receptors, did not exert any behavioural effects in either the 5-, 10- or 20-day-old rat pups at doses of up to 10 mg/kg. These results demonstrate behavioural effects of - and -but not -agonists in neonatal rats. There is a clear differentiation between - and -receptor effects and both - and -mediated behaviours show dissimilarities from the adult profile.     

Interactions of three inotropic agents,ASL-7022, dobutamine and dopamine,with α- and β-adrenoceptors in vitro

Summary Three inotropic agents, ASL-7022, dobutamine and dopamine, were evaluated for their -and -adrenoceptor mediated effects in vitro in a variety of isolated organs and in radioligand binding studies. All compounds were ₁-adrenoreceptor agonists in rat and guinea pig aortae, but the rank orders of potency were exactly opposite in these two tissues. Only the rank potency order of dobutamine>ASL-7022>dopamine obtained in rat aorta was consistent with the results obtained in radioligand binding studies to ₁-adrenoreceptors in rat cerebral cortex and to previous results obtained in vivo in the pithed rat. The results obtained in guinea pig aorta did not parallel the radioligand binding studies in rat brain or our previous results in pithed rat, and suggests that species differences exist between postsynaptic vascular ₁-adrenoreceptors in rat and guinea pig aorta, consistent with previous conclusions. ASL-7022 was found to be a potent ₂-adrenoreceptor agonist in field-stimulated guinea pig ileum, and was approximately 10-fold more potent than dobutamine in this respects, which was also confirmed by radioligand binding studies to ₂-adrenoreceptors in rat cerebral cortex. The ₁-adrenoreceptor mediated effects of these compounds were evaluated in guinea pig atria, where the rank order of potency was dobutamine>ASL-7022>dopamine. An identical rank order of affinity was established for these compounds by displacement of ³H-dihydroalprenolol from ₁-adrenoreceptors in rat cerebral cortex. The ₁-adrenoreceptor mediated effects of dobutamine and ASL-7022 in guinea pig atria were completely direct in nature and not secondary to the release of endogenous catecholamines. In contrast, a major component of the ₁-adrenoreceptor mediated tachycardia produced by dopamine in guinea pig atria was indirect in nature as evidenced by the marked attenuation in potency that occurred following catecholamine depletion with reserpine. All three compounds elicited ₂-adrenoreceptor mediated inhibition of tone in rat uterus, with the rank order of potency being ASL-7022>dobutamine>dopamine. Again, this rank order of ₂-adrenoreceptor potency was also reflected in ₂-adrenoreceptor affinity as assessed by displacement of ³H-dihydroalprenolol from ₂-adrenoreceptors in rat cerebellum. Based on these results, it may be concluded that for -adrenoceptors, dobutamine is a selective ₂-adrenoreceptor agonist, ASL-7022 is a selective ₂-adrenoreceptor agonist, and dopamine is a nonselective -adrenoceptor agonist. For -adrenoceptor mediated effect, ASL-7022 is a selective ₂ agonist, while dobutamine and dopamine are nonselective -adrenoceptor agonists. It is likely that the complex inotropic and hemodynamic activities of ASL-7022, dobutamine and dopamine result from the sum of their individual effects at the -and -adrenoceptor subtypes.     

Metabolic fate of sipoglitazar, a novel oral PPAR agonist with activities for PPAR-γ, -α and -δ, in rats and monkeys and comparison with humans in vitro

Sipoglitazar is a novel anti-diabetic agent with triple agonistic activities on the human peroxisome proliferator-activated receptors, hPPAR-γ, -α, and -δ. The bioavailability for sipoglitazar was 95.0% and 72.6% in rats and monkeys respectively and sipoglitazar is hardly subject to first pass metabolism in either species. Following oral administration of [1?C]sipoglitazar to rats, sipoglitazar and its metabolites were distributed to the rat tissues with relatively high concentrations in the liver and also to the target tissue, the adipose tissue. The major component was sipoglitazar in the plasma of rats and monkeys. In rats, sipoglitazar was mainly excreted into the feces via biliary excretion as sipoglitazar-G, while the major component was M-I-G in the urine and M-I in the feces of monkeys. In hepatocytes, the metabolism was not extensively advanced in rats and the main metabolites were M-I and sipoglitazar-G in humans, similar to the metabolic profile in monkeys. There was no metabolite specific for humans in vitro. In conclusion, the formation of M-I, M-I-G and sipoglitazar-G is considered to be crucial and sipoglitazar is presumed to be cleared primarily by oxidation and glucuronidation in humans, when examined in vivo and in vitro.     

Association of genes coding for the α-4, α-5, β-2 and β-3 subunits of nicotinic receptors with cigarette smoking and nicotine dependence

We assessed whether smoking behavior was associated with nine polymorphisms in genes coding for the nicotinic receptor subunits α-4 (rs1044394, rs1044396, rs2236196 and rs2273504), α-5 (rs16969968), β-2 (rs2072661 and rs4845378) and β-3 (rs4953 and rs6474413). We conducted an Internet survey and collected saliva by mail for DNA and cotinine analyses, in Switzerland in 2003. We conducted DNA analyses for 277 participants and cotinine analyses for 141 current daily smokers. Cotinine levels were higher in carriers of the CC genotype of CHRNA4 rs1044396 (371 ng/ml) than in those with the CT or TT genotypes (275 ng/ml, p = 0.049), a difference of 0.53 standard deviation units. However, this difference was not robust to correction for multiple testing using Bonferroni adjustment. These 9 polymorphisms were not otherwise associated with smoking behavior and nicotine dependence. There were possible associations between the temperament trait novelty seeking and CHRNA4 rs1044396, CHRNA5 rs16969968 and CHRNB2 rs4845378, but these associations were not robust to correction for multiple testing. We conclude that the analysis of polymorphisms in genes coding for four nicotinic acetylcholine receptor subunits (CHRNA4, CHRNA5, CHRNB2 and CHRNB3) and several smoking-related phenotypes revealed no statistically significant association.     

Alzheimer's disease, β-amyloid, glutamate, NMDA receptors and memantine - searching for the connections

β-amyloid (Aβ) is widely accepted to be one of the major pathomechanisms underlying Alzheimer's disease (AD), although there is presently lively debate regarding the relative roles of particular species/forms of this peptide. Most recent evidence indicates that soluble oligomers rather than plaques are the major cause of synaptic dysfunction and ultimately neurodegeneration. Soluble oligomeric Aβ has been shown to interact with several proteins, for example glutamatergic receptors of the NMDA type and proteins responsible for maintaining glutamate homeostasis such as uptake and release. As NMDA receptors are critically involved in neuronal plasticity including learning and memory, we felt that it would be valuable to provide an up to date review of the evidence connecting Aβ to these receptors and related neuronal plasticity. Strong support for the clinical relevance of such interactions is provided by the NMDA receptor antagonist memantine. This substance is the only NMDA receptor antagonist used clinically in the treatment of AD and therefore offers an excellent tool to facilitate translational extrapolations from in vitro studies through in vivo animal experiments to its ultimate clinical utility.     

Study of the genotoxic and cytotoxic effects of the α-, β-, and γ- Hexachlorocyclohexane isomers in human lymphocyte cells using the cytokinesis-block micronucleus assay

The genotoxic potential of hexachlorocyclohexane (HCH) isomers (α-, β-, and γ-) which are organochlorine pesticides was tested in peripheral blood lymphocyte cultures from two donors by using the cytokinesis-block micronucleus assay. Micronucleus (MN) frequency, binucleated cells with micronucleus (BNMN), and cytokinesis-blocked proliferation index (CBPI) were determined as genotoxic and cytotoxic endpoints. At the concentration ranges tested (12.5–100?μg.L?^?1), all HCH isomers induced dose-dependent cytotoxic effects, γ-HCH being the most toxic. This isomer was also able to induce significant increase in MN frequency and BNMN cells indicating a genotoxic potential at 50 and 100?μg.L?^?1. The genotoxic test of β-HCH showed a positive induction of MN and BNMN cells at the highest concentration of 100?μg.L?^?1 and a significant cytotoxicity at 50?μg.L?^?1. Under the experimental condition used, α-HCH was unable to induce any significant increase in MN frequency confirming that α-HCH is a non-genotoxic agent.     

Effects of μ-, δ- and κ-opioid antagonists in atrial preparations from nonfailing and failing human hearts

• 1.1. We examined the effects of naloxone (preferentially μ-antagonist), naltrindole (selective δ-antagonist) or nor-binaltorphimine (nor-BNI, selective κ-antagonist) on auricular myocardium tissue from nonfailing and failing human hearts.
• 2.2. The opioid antagonists used in this study induced inhibitory effects in auricular strips from failing and nonfailing human hearts. In addition, the maximal effect, the IC₅₀, and the slope of concentration-response curves obtained with μ-, δ-, and κ-opioid antagonists were similar in failing and nonfailing human heart tissues.
• 3.3. The K-antagonist was more effective than naltrindole or naloxone. Moreover, the IC₅₀ for nor-BNI (0.25±0.01 × 10⁻⁵M) was lower than the IC₅₀ for naloxone (26.5±5.0 × 10⁻⁵M) and naltrindole (13.8±2.0 × 10⁻⁵M). Similar results were obtained in auricular strips from failing human hearts.
• 4.4. Our results demonstrate that the failing heart does not modify the inhibitory cardiac effects obtained with selective opioid antagonists.

     

Preparation,release and physicochemical characterisation of ethyl butyrate and hexanal inclusion complexes with β- and γ-cyclodextrin

Abstract

Complexes of ethyl butyrate and hexanal encapsulated by β-cyclodextrin (β-CD) and γ-cyclodextrin (γ-CD) were prepared by coprecipitation, and gas chromatography was used to quantity the flavour compounds in the complexes. The ethyl butyrate–γ-CD complex had the highest inclusion ratio (12.20%) followed by the ethyl butyrate-β-CD, hexanal-β-CD and hexanal-γ-CD complexes (11.29, 4.41 and 3.33%, respectively). Release experiments were performed under different relative humidities (RH 93, 75 and 52%) and temperatures (4 and 25?°C). The flavour release behaviours of the complexes were described by the Avrami equation. The rate of flavour release was enhanced with both increasing temperature and RH, although the effect of RH was stronger. Physicochemical characterisation using FT-IR, XRD, DSC and SEM analyses demonstrated that crystalline complexes were formed. Both β-CD and γ-CD were able to encapsulate ethyl butyrate and hexanal, and lower RH and temperature were more suitable for the storage of these complexes.