Uptake and localisation of O-(β-hydroxyethyl)-rutosides in the venous wall,measured by laser scanning microscopy

Summary The uptake and localisation of O-(-hy-droxyethyl)-rutosides (HR) in the venous wall was studied in 8 patients undergoing crossectomy for a varicose long saphenous vein. The fluorescence of cross-sections of the vein wall was measured by laser scanning microscopy, based on the autofluorescence of HR. Four patients (treated group) received 2 × 1.5 g HR IV before surgery, and four (untreated patients) served as controls.Uptake of HR into the veins from treated patients was seen, with a mean fluorescence intensity of 80.9 units compared to 49.4 units in the untreated veins. The increase in fluorescence was clearly demarcated on the endothelial side of the vein wall.It is concluded that HR passes into the vascular wall, where it is localised in the endothelial and sub-endothelial areas.     

Time course of the anti-oedematous effect of O-(β-hydroxyethyl)-rutosides in healthy volunteers

Summary O-(-hydroxyethyl)-rutosides (HR) is used for the treatment of disorders of the venous and microcirculatory systems. In order to evaluate the time course of its activity, the effect of HR on a provocation model of orthostatic oedema in healthy volunteers was used. After a 2 week placebo run-in period, 16 healthy volunteers were randomized to HR (2 tablets of 500 mg/day) of placebo for a further 3 weeks, in a double-blind parallel design. Oedema was provoked by standing motionless for 1 h, with measurement of leg volume before and afterwards. The procedure was undertaken at entry to the study and then weekly during the entire 5 week period.There were no significant differences in the extent of oedema produced by the orthostatic challenge during the 2 week run-in period or in the subjects who continued on placebo (90 arbitrary units i.e. 48 ml). During the 3 week treatment with HR, however, there was a progressive reduction (–1.1, –5.9, and –7.6 arbitrary units after 1, 2, and 3 weeks, respectively) in the volume of induced oedema, which was significant after 2 and 3 weeks of treatment compared to the placebo group.     

Polymorphs and permeability of 3',4'-dimethoxy flavonol-3-O-β-D-glucopyranoside monohydrate

3',4'-Dimethoxy-flavonol-3-O-β-D-glucopyranoside monohydrate (GDH), which can significantly reduce blood lipids, atherosclerotic aortic lesions, and liver injury, has poor oral bioavailability. In the present study, we aimed to prepare and characterize five new polymorphs of GDH (II, III, IV, V, and VI) and the amorphous form of GDH (GDH-AM). The GDH polymorphs and GDH-AM were characterized by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray powder diffraction (XRPD), and Fourier transform infrared spectroscopy (FTIR). Dissolution tests, physical stability, polymorphic transformation, and permeability studies were subsequently investigated. Dissolution of GDH-II and GDH-IV reached higher concentrations compared with GDH-I. GDH-AM exhibited a significantly high dissolution rate and prolonged supersaturation during dissolution. No phase transition was found for GDH-I and GDH-AM after 3 months of storage, while GDH-II, GDH-III, GDH-IV, GDH-V, and GDH-VI were readily converted to GDH-I. In situ single-pass intestinal perfusion experiments showed that the high concentration of GDH exhibited low permeability. Sodium dodecyl sulfate and bovine bile salts were used as absorption enhancers to improve the permeability of GDH. The results showed that sodium dodecyl sulfate and taurocholate were good absorption enhancers for further formulation development of GDH.     

The Disposition of 10–10′ Oxybisphenoxarsine (OBPA) in Rats,Rabbits and Guinea Pigs

ABSTRACT

The disposition of 10–10’ oxybisphenoxarsine (OBPA), a potent, wide spectrum anti-microbial agent widely used in plastics, was investigated in rats, guinea pigs and rabbits. All animals were given a single dose of [U- ¹⁴C]OBPA by gavage. Urine and feces were collected daily for 7 days, at which time the animals were sacrificed and tissue samples removed for analysis. In all three species, approximately 90% of the administered dose was excreted within 7 days. The rabbits and guinea pigs excreted about 58% via the feces and 32% via the urine. The rats, however, excreted 82% via the feces and 10% via the urine. There were significant differences in the tissue distribution of radioactivity between the three species. The most prominent difference was that the erythrocytes of the rat retained 3.2% of the administered radiolabel while only 0.08% and 0.03% of the dose remained in the erythrocytes of the guinea pig and rabbit, respectively.     

Studies on the Metabolism of Sympathomimetic Amines. The Metabolism of (±)-[14C]Noradrenaline in the Horse

Abstract

1. The metabolism of (±)-[¹⁴C]noradrenaline in horses has been studied. The plasma half-life of radioactivity following intravenous injection was 95 min.

2. Two horses each excreted about 80–85% of the radioactivity in the urine in 15 h after rapid intravenous injection and about 75% of the excreted radioactivity has been identified.

3. The unchanged drug in the urine accounted for less than 1% of the dose and 3-methoxynoradrenaline for about 7%. The main metabolites were 4-hydroxy-3-methoxymandelic acid (22%), 4-hydroxy-3-methoxybenzoic acid (13%) and 4-hydroxy-3-methoxyphenylglycol (11%). 3,4-Dihydroxyphenylglycol was a minor metabolite (5%).     

Studies on the Metabolism of Sympathomimetic Amines. The Metabolism of (±)-[14C] Adrenaline in the Horse

Abstract

1. The metabolism of (±)-[¹⁴C]adrenaline has been studied in two horses after intravenous administration.

2. The plasma half-life of ¹⁴C was 228 min (Horse A) and 100 min (Horse B).

3. Excretion of ¹⁴C in the urine in 20 h was 90% (A) and 70% (B). Over 94% of the excreted radioactivity has been identified.

4. The two horses differed in their metabolism of adrenaline. Unchanged drug accounted for 11% (A) and 20% (B), and for A and B respectively, metabolites excreted were 3-methoxyadrenaline (10%, 21%), 4-hydroxy-3-methoxymandelic acid (16%, 13%), 3,4-dihydroxyphenylglycol (6%, 0%), 4-hydroxy-3-methoxyphenylglycol (10%, 3%), 3,4-dihydroxybenzoic acid (5%, 0%), 3,4-dihydroxymandelic acid (7%, 3%) and 4-hydroxy-3-methoxybenzoic acid (18%, 4%).     

The syntheses and characterization 3β-(4-fluorobenzoyloxy)tropane (fluorotropacocaine) and its 3α isomer

3β‐(4‐Fluorobenzoyloxy)tropane (3β‐FBT, fluorotropacocaine) was first reported by Finnish authorities to the European Monitoring Centre for Drugs and Drug Addiction (EMCDDA) via the Early Warning System (EWS) in 2008 and our own laboratory tentatively identified it in 2010 in several products purchased from head shops. Very little is known about this cocaine‐like drug and, as no reference standards were available, we have synthesized and characterized both 3β‐FBT and its 3α isomer for use as reference standards. The two compounds are separable by gas chromatography (GC) but their electron‐impact (EI) mass spectra were found to be almost identical. ¹⁹F NMR spectroscopy was also found to be a useful technique for distinguishing the two isomers. Copyright © 2011 John Wiley & Sons, Ltd.     

Metabolism in vivo of 3,4,3′,4′-tetrachlorobiphenyl and toxicological assessment of the metabolites in rats

1. Metabolism in vivo of 3,4,3′,4′-tetrachlorobiphenyl (TCB) and toxicological assessment of the metabolites were investigated in the rat.

2. Four metabolites were isolated from faeces of rats dosed with 3,4,3′,4′-TCB. Two were identified as 5-hydroxy-3,4,3′,4′-TCB and a chlorine-shift metabolite, 4-hydroxy-3,5,3′,4′-TCB, by comparison of melting points, chromatographic mobilities and spectral features with those of the synthetic samples. A dihydroxy-TCB and monohydroxy-trichlorobiphenyl were also indicated by mass spectrometry to be excreted in faeces as minor metabolites.

3. Faecal excretion of unchanged 3,4,3′,4′-TCB, 5-hydroxy-3,4,3′,4′-TCB and 4-hydroxy-3,5,3′,4′-TCB was 0.8%, 19.6% and 11.6% of dose, respectively, in 5 days after i.p. injection of 3,4,3′,4′-TCB at a dose of 50mg/kg.

4. From the inability to cause the liver hypertrophy and thymus atrophy, both monohydroxy-metabolites of 3,4,3′,4′-TCB are much less toxic than the parent 3,4,3′,4,-TCB. In addition, these phenolic metabolites did not induce the activities of benzo[a]pyrene hydroxylase and DT-diaphorase, whereas 3,4,3′,4′-TCB greatly induced these activities. These results indicated that unlike PCB congeners with phenobarbital-type inducing ability, 3,4,3′,4′-TCB, a prototype of 3-methylcholanthrene-type inducers. is detoxified by metabolic hydroxylation.     

The utility of endogenous glycochenodeoxycholate-3-sulfate and 4β-hydroxycholesterol to evaluate the hepatic disposition of atorvastatin in rats

The liver is an important organ for drugs disposition, and thus how to accurately evaluate hepatic clearance is essential for proper drug dosing. However, there are many limitations in drug dosage adjustment based on liver function and pharmacogenomic testing. In this study, we evaluated the ability of endogenous glycochenodeoxycholate-3-sulfate (GCDCA-S) and 4β-hydroxycholesterol (4β-HC) plasma levels to evaluate organic anion-transporting polypeptide (Oatps)-mediated hepatic uptake and Cyp3a-meidated metabolism of atorvastatin (ATV) in rats. The concentration of ATV and its metabolites, 2-OH ATV and 4-OH ATV, was markedly increased after a single injection of rifampicin (RIF), an inhibitor of Oatps. Concurrently, plasma GCDCA-S levels were also elevated. After a single injection of the Cyp3a inhibitor ketoconazole (KTZ), plasma ATV concentrations were significantly increased and 2-OH ATV concentrations were decreased, consistent with the metabolism of ATV by Cyp3a. However, plasma 4β-HC was not affected by KTZ treatment despite it being a Cyp3a metabolite of cholesterol. After repeated oral administration of RIF, plasma concentrations of ATV, 2-OH ATV and 4-OH ATV were markedly increased and the hepatic uptake ratio of ATV and GCDCA-S was decreased. KTZ did not affect plasma concentrations of ATV, 2-OH ATV and 4-OH ATV, but significantly decreased the metabolic ratio of total and 4-OH ATV. However, the plasma level and hepatic metabolism of 4β-HC were not changed by KTZ. The inhibition of hepatic uptake of GCDCA-S by RIF was fully reversed after a 7-d washout of RIF. Plasma concentration and hepatic uptake ratio of GCDCA-S were correlated with the plasma level and hepatic uptake of ATV in rats with ANIT-induced liver injury, respectively. These results demonstrate that plasma GCDCA-S is a sensitive probe for the assessment of Oatps-mediated hepatic uptake of ATV. However, Cyp3a-mediated metabolism of ATV was not predicted by plasma 4β-HC levels in rats.     

The Efflux of Flavonoids Morin, Isorhamnetin-3-O-Rutinoside and Diosmetin-7-O-β-d-Xylopyranosyl-(1-6)-β-d-Glucopyranoside in the Human Intestinal Cell Line Caco-2

Purpose In this study, we chose three of the flavonoids isorhamnetin-3-O-rutinoside(IRR) diosmetin-7-O-β-d-xylopyranosyl-(1-6)-β-d-glucopyranoside(DXG) and morin, which showed obvious efflux, to test the hypothesis that a specific efflux transporter is responsible for their transportation.Methods The intestinal epithelial membrane transport of the flavonoids were examined using the monolayer of the human Caco-2 cell line grown in Transwells, a common model of intestinal absorption. The flavonoids were measured by high performance liquid chromatography with UV detector.Result The efflux of morin, IRR and DXG, across Caco-2 cell monolayers was examined over the concentration range from 2 to 200 μM and showed a saturable process. The depletion of the cellular ATP stores with 5 mM iodoacetamide led to a significant inhibition of the efflux. Fifty micromolar verapamil, a chemical inhibitor of P-glycoprotein, had no effect on the transport of the three flavonoids, while the presence of 50 μM MK-571 and 1 mM probenecid, MRP inhibitors, resulted in an obvious reduction in the efflux. Moreover, inhibition of morin transport by MK-571 demonstrated concentration dependence. The transportation of the three flavonoids was compared with apigenin.Conclusion These data support a role for MRPs in the intestinal transcellular efflux of morin, IRR, DXG and possibly other hydrophilic flavonoid aglycons and glycosides.     

Hepatic Metabolism and Disposition of Baicalein via the Coupling of Conjugation Enzymes and Transporters—In Vitro and In Vivo Evidences

Baicalein (Ba) was found to be subject to serious first-pass metabolism after oral administration. We previously revealed the important role of intestine in the low oral bioavailability of Ba. The present study aims to evaluate the hepatic metabolism and disposition of Ba. Ba was given to Sprague-Dawley rats through bolus or infusion via intravenous or intra-portal route of administrations. Both plasma and bile samples at different time intervals were obtained. Concentrations of Ba and potential metabolites in the collected samples were analyzed with HPLC/UV and identified by LC/MS/MS, respectively. Plasma concentration versus time profiles of Ba obtained from intravenous and intra-portal administrations were compared to estimate the extent of hepatic metabolism. In addition, transport studies of baicalein-7-glucuronide (BG), one of the major metabolites of Ba, were carried out using transfected cell systems overexpressing various human organic anion-transporting polypeptide (OATP) isoforms to estimate the specific transporters involved in the hepatic disposition of Ba metabolites. The results showed that liver, in addition to intestine, also conferred extensive metabolism to Ba. Several mono- and di-conjugates of Ba, which were mainly glucuronides, sulfates, and methylates, were found in bile. The transport study demonstrated that besides MRPs and BCRP, human OATP2B1 and OATP1B3 in liver might also mediate the secretion of BG to bile. In summary, liver plays an important role in the metabolism of Ba and transport of its conjugated metabolites.     

The Metabolism of the Neuroleptic Agent 1-(4′ -Fluorophenyl)-4-(cyclohexyl-1′-piperazinyl-4′-carboxylate)-butan-1-one Hydrochloride in Rats and Man

1. An oral dose of the neuroleptic agent 1-(4′-fluor ophenyl)-4-(cyclohexyl-1′-[¹⁴ C]piperazinyl-4′-carboxylate)butan-1-one was mainly eliminated in the urine within 12?h by rats and man. During 5 days, 63.6% and 83.3% was eliminated in the urine of rats and man respectively.

2. Plasma concentrations in man reached a maximum during 30 min to 1?h, representing 1.43μg equiv./ml. The proportion of unchanged drug in plasma decreased from 48% at 15 min to less than 10% after 1 h.

3. Seven major radioactive components were detected in the chloroform extract of basified rat urine and five major components in similar extracts of human urine. The major rat metabolites were isolated and identified by mass spectrometry as components resulting from mono- and dihydroxylation in the cyclohexane ring, reduction of the keto group to a secondary alcohol and hydrolysis and decarboxylation of the cyclohexylcarbamoyl group. The major metabolite in the rat urine extract was the dihydroxylated secondary alcohol derivative while the major human metabolite was the monohydroxylated secondary alcohol derivative. The metabolites were also partly eliminated as conjugates.     

Quantitative Evaluation of Ethanol Effects on Diffusion and Metabolism of β-Estradiol in Hairless Mouse Skin

The influence of low levels of ethanol on the simultaneous diffusion and metabolism of -estradiol (E₂) in hairless mouse skin was quantitatively evaluated. A wide range of diffusion/metabolism experiments was conducted with full-thickness skin, stripped skin, and dermis at the various ethanol levels. The experiments were carried out in a two-chamber diffusion-cell system where ethanol was present in both the donor and the receiver chambers at equal concentrations. Analysis of the experimental data with several enzyme distribution models further showed that the best model was that for which the enzyme activity resided totally in the epidermis and near the basal layer of the epidermis. The ethanol effects were separated and quantified in terms of the diffusion and metabolism parameters. Aqueous ethanol, even at low concentrations (25%), was found to have two important effects on E₂ transport: ethanol functions as an inhibitor of the enzymatic conversion of E₂ to estrone (E₁) in the viable epidermis, and ethanol is able to enhance the transport of permeants across the lipoidal pathway of the stratum corneum.     

The expression and clinical significance of P-GSK3βandβ-catenin in endometrial carcinoma

目的 检测子宫内膜癌组织中P-GSK3β蛋白和β-catenin蛋白表达并探讨其意义.方法 采用微波EliVisionTM免疫组织化学法检测60例子宫内膜癌,32例正常子宫内膜组织中P-GSK3β蛋白和β-catenin蛋白的表达.结果 正常子宫内膜组织中β-catenin蛋白表达正常,子宫内膜癌组织中β-catenin发生异位表达.子宫内膜癌组织中P-GSK3β和异位表达的β-catenin分别定位于细胞浆和细胞核,两者表达明显高于癌旁正常组织(P<0.05).两者的表达与肌层浸润程度、淋巴结转移、临床分期及病理分期有关 (P<0.05);两者之间表达呈正相关(P<0.05).结论 P-GSK3β表达和β-catenin异位表达在子宫内膜癌呈高表达,且与子宫内膜癌的发生、发展和转移密切相关.     

Further analyses of mechanisms underlying the antinociceptive effect of the triterpene 3β, 6β, 16β-trihydroxylup-20(29)-ene in mice

As a critical component of post-resuscitation care, prompt optimization of hemodynamic status by means of targeted interventions is vital in order to maximize the likelihood of good outcome. Vasoactive agents play an essential role in the supportive care of post cardiac arrest patients. The administration of these agents is associated with serious side-effects and therefore they should be used in the minimal dose necessary to achieve low-normal mean arterial pressure and adequate systematic perfusion. Careful and frequent serial evaluation of the patient is important primarily to assess volume status and adequacy of circulatory support. Continuous monitoring of blood pressure and laboratory parameters is essential both to accurately titrate therapy and because inotropes and vasopressors have the potential to induce life-threatening side-effects. The clinical efficacy of inotropes and vasopressors has been largely investigated through examination of their impact on hemodynamic end points, and clinical practice has been driven in part by expert opinion, extrapolation from animal studies, and physician preference. Clearly these agents should all be considered as supportive measures to stabilize the patient prior to some form of definitive therapy.     

Metabolism of Linoleamides: II. Absorption,Distribution, Excretion and Metabolism of N-[α-Phenyl-β-(p-tolyl)ethyl] linoleamide

1. Absorption, distribution, excretion and metabolism of (-)N-[α-phenyl-β-(p-tolyl)ethyl][¹⁴C]linoleamide (¹⁴C-PTLA) were studied in rats and dogs. Faecal excretion of PTLA was studied in dogs and men by g.l.c.

2. ¹⁴C-PTLA (10 mg/kg) given orally to rats resulted in urinary and faecal excretion of radioactivity of 2 and 93 %, respectively, by male rats and 8 and 87% by female rats in 48 h. Faecal excretion of PTLA in men was similar to that in rats.

3. Distribution of radioactivity in rats and dogs after oral administration of ¹⁴C-PTLA showed that a major part of the dose was not absorbed.

4. N-[α-Phenyl-β-(p-tolyl)ethyl]succinic acid monoamide and N-[α-phenyl-β-(p-tolyl)ethyl]glutaric acid monoamide were detected in the urine of rats dosed orally with ¹⁴C-PTLA.     

Effects of the specific α4β2 nAChR antagonist, 2-fluoro-3-(4-nitrophenyl) deschloroepibatidine, on nicotine reward-related behaviors in rats and mice

Rationale

Alleviating addiction to tobacco products could prevent millions of deaths. Investigating novel compounds selectively targeting α4β2 nAChRs hypothesized to have a key role in the rewarding effects of nicotine may be a useful approach for future treatment.

Objectives

The present study was designed to evaluate 2-fluoro-3-(4-nitrophenyl) deschloroepibatidine (4-nitro-PFEB), a potent competitive antagonist of neuronal α4β2 nAChRs, in several animal models related to nicotine reward: drug discrimination, intracranial self-stimulation (ICSS), conditioned place preference, and limited access to self-administration.

Methods

Long Evans rats were trained in a two-lever discrimination procedure to discriminate 0.4 mg/kg nicotine (s.c.) from saline. Male Sprague–Dawley rats were stereotaxically implanted with electrodes and trained to respond for direct electrical stimulation of the medial forebrain bundle. ICR mice were evaluated using an unbiased place preference paradigm, and finally, male Wistar rats were implanted with intrajugular catheters and tested for nicotine self-administration under limited access (1 h/day).

Results

4-Nitro-PFEB attenuated the discriminative stimulus effects of nicotine, but alone did not produce nicotine-like discriminative stimulus effects. Nicotine-induced facilitation of ICSS reward thresholds was reversed by 4-nitro-PFEB, which alone had no effect on thresholds. 4-Nitro-PFEB also blocked the conditioned place preference produced by nicotine, but alone had no effect on conditioned place preference. Finally, 4-nitro-PFEB dose-dependently decreased nicotine self-administration.

Conclusions

These results support the hypothesis that neuronal α4β2 nAChRs play a key role in mediating the rewarding effects of nicotine and further suggest that targeting α4β2 nAChRs may yield a potential candidate for the treatment of nicotine dependence.     

Synthesis and cytotoxicity of (3β)-3-acetyloxy-5(6)-androsten-7-one oxime and 3,5(6)-androstadien-7-one oxime

(3β)-3-Acetyloxy-5(6)-androsten-7-one oxime and 3,5(6)-androstadien-7-one oxime were synthesized and their in vitro cytotoxic activities against human epidermoid carcinoma cell line KB, human cervical carcinoma cell line HeLa, human gastric carcinoma cell line MKN-28, and human breast carcinoma cell line MCF-7 were evaluated. The compound (3β)-3-acetyloxy-5(6)-androsten-7-one oxime exhibited significant growth inhibitory properties against cell lines tested and afforded IC₅₀ value 26.0 ± 1.6, 12.8 ± 1.0, 18.1 ± 0.2, 10.2 ± 1.2 μM for KB, HeLa, MKN-28, and MCF-7, respectively.     

Section 3 The tolerance and acceptability of ibuprofen (‘Brufen’) in the elderly patient

Summary

Data on the treatment of patients over the age of 65 years were extracted from the computer records of a long-term multi-centre study of ibuprofen. Patients were suffering from rheumatoid disease, osteoarthrosis or an allied condition. Records from 231 patients were available for analysis. Duration of therapy with ibuprofen ranged from 3 months to over 5 years. Assessments of clinical progress, as judged by both patients and physicians, changes in biochemical and haematological parameters, and side-effects showed that ibuprofen is an effective form of therapy and is generally well tolerated in the elderly.