Comparison of analgesic effects and patient tolerability of nabilone and dihydrocodeine for chronic neuropathic pain: randomised,crossover, double blind study

Objective To compare the analgesic efficacy and side effects of the synthetic cannabinoid nabilone with those of the weak opioid dihydrocodeine for chronic neuropathic pain. Design Randomised, double blind, crossover trial of 14 weeks’ duration comparing dihydrocodeine and nabilone.Setting Outpatient units of three hospitals in the United Kingdom.Participants 96 patients with chronic neuropathic pain, aged 23-84 years. Main outcome measures The primary outcome was difference between nabilone and dihydrocodeine in pain, as measured by the mean visual analogue score computed over the last 2 weeks of each treatment period. Secondary outcomes were changes in mood, quality of life, sleep, and psychometric function. Side effects were measured by a questionnaire.Intervention Patients received a maximum daily dose of 240 mg dihydrocodeine or 2 mg nabilone at the end of each escalating treatment period of 6 weeks. Treatment periods were separated by a 2 week washout period.Results Mean baseline visual analogue score was 69.6 mm (range 29.4-95.2) on a 0-100 mm scale. 73 patients were included in the available case analysis and 64 patients in the per protocol analysis. The mean score was 6.0 mm longer for nabilone than for dihydrocodeine (95% confidence interval 1.4 to 10.5) in the available case analysis and 5.6 mm (10.3 to 0.8) in the per protocol analysis. Side effects were more frequent with nabilone.Conclusion Dihydrocodeine provided better pain relief than the synthetic cannabinoid nabilone and had slightly fewer side effects, although no major adverse events occurred for either drug. Trial registration Current Controlled Trials ISRCTN15330757.     

Microbiological transformations of nabilone, a synthetic cannabinoid.

A screening program was conducted to find microorganisms that modify the synthetic cannabinoid nabilone. After purification, the products from three cultures were analyzed by spectral methods to determine their chemical structures. An optically active 9S-hydroxy-6aR,10aR-trans cannabinoid was isolated from a culture of an unidentified soil bacterium designated A24007. From Bacillus cereus cultures were isolated a 9S,6'-dihydroxy-6aR,10aR-trans cannabinoid, a 9S-hydroxy-6'-keto-6aR,10aR-trans cannabinoid, a 9-keto-6'-hydroxy-6aS,10aS-trans cannabinoid, and a 6',9-diketo-6aS,10aS-trans cannabinoid. All of these products were optically active, as was a 9S-hydroxy-6aS,10AS-trans cannabinoid also isolated from B. cereus cultures. A series of acidic products were isolated from cultures of Nocardia salmonicolor. All of these products contained a carboxylic acid group at the terminal end of three-position alkyl side chains having varying numbers of carbon atoms. Two of the acidic products contained a 9-keto group, whereas all other carboxylic acid products were 9-hydroxy cannabinoids. The array of products obtained from incubation of nabilone indicates the usefulness of microbial transformations in the preparation of new cannabinoids.     

Wounding of Arabidopsis leaves induces indole‐3‐carbinol‐dependent autophagy in roots of Arabidopsis thaliana

In cruciferous plants insect attack or physical damage induce the synthesis of the glucosinolate breakdown product indole‐3‐carbinol, which plays a key role in the defense against attackers. Indole‐3‐carbinol also affects plant growth and development, acting as an auxin antagonist by binding to the TIR1 auxin receptor. Other potential functions of indole‐3‐carbinol and the underlying mechanisms in plant biology are unknown. Here we show that an indole‐3‐carbinol‐dependent signal induces specific autophagy in root cells. Leaf treatment with exogenous indole‐3‐carbinol or leaf‐wounding induced autophagy and inhibited auxin response in the root. This induction is lost in glucosinolate‐defective mutants, indicating that the effect of indole‐3‐carbinol is transported in the plants. Thus, indole‐3‐carbinol is not only a defensive metabolite that repels insects, but is also involved in long‐distance communication regulating growth and development in plants.     

The glucosinolate breakdown product indole‐3‐carbinol acts as an auxin antagonist in roots of Arabidopsis thaliana

The glucosinolate breakdown product indole‐3‐carbinol functions in cruciferous vegetables as a protective agent against foraging insects. While the toxic and deterrent effects of glucosinolate breakdown on herbivores and pathogens have been studied extensively, the secondary responses that are induced in the plant by indole‐3‐carbinol remain relatively uninvestigated. Here we examined the hypothesis that indole‐3‐carbinol plays a role in influencing plant growth and development by manipulating auxin signaling. We show that indole‐3‐carbinol rapidly and reversibly inhibits root elongation in a dose‐dependent manner, and that this inhibition is accompanied by a loss of auxin activity in the root meristem. A direct interaction between indole‐3‐carbinol and the auxin perception machinery was suggested, as application of indole‐3‐carbinol rescues auxin‐induced root phenotypes. In vitro and yeast‐based protein interaction studies showed that indole‐3‐carbinol perturbs the auxin‐dependent interaction of Transport Inhibitor Response (TIR1) with auxin/3‐indoleacetic acid (Aux/IAAs) proteins, further supporting the possibility that indole‐3‐carbinol acts as an auxin antagonist. The results indicate that chemicals whose production is induced by herbivory, such as indole‐3‐carbinol, function not only to repel herbivores, but also as signaling molecules that directly compete with auxin to fine tune plant growth and development.     

Formation of some monopositive and dipositive carbonium ions in chlorosulphuric acid. Part XV

Tetraphenyl-p-xylene-glycol, tetraphenyl-phthalein and Dipheno(3-10′)thiazinyl are shown to form dipositive carbonium ions, but triphenyl acetic acid, 2-3-5-6 tetramethyl benzoic acid, 2-3-4-5-6 pentamethyl benzoic acid, tert-butyl alcohol, triphenyl carbinol, tri-p-tolyl carbinol, tri-o-tolyl carbinol, tri-p-chlorophenyl carbinol and tri-p-nitrophenyl carbinol form monopositive carbonium ions in chlorosulphuric acid, as revealed by conductometric and u.v. spectral studies. Oxalyl chloride decomposes while ethylene glycol is sulphonated in chlorosulphuric acid. Dichloroethane behaves as a non-electrolyte but dibromomethane disproportionates in this medium.     

New labdane derivatives and further constituents of Brickellia species

The investigation of several Brickellia species and two further related species afford besides known compounds four new labdane derivatives, a new acetylenic carbinol and a new dihydrobenzofuran derivative. The structures have been elucidated by spectroscopic methods. The chemotaxonomic aspects are discussed briefly.     

甲醇对酵母过氧化氢酶活性的影响机理研究

将酵母过氧化氢酶加入一定比例的甲醇,测定其活性变化。结果表明:在含2%甲醇时酶活比对照提高4026%。将粗酶液用70%饱和度的硫酸铵盐析后离心所得的上清液再加入硫酸铵至80%饱和度,离心的沉淀溶解在缓冲液中,上SephadexG75柱,分离出的有酶活性的蛋白峰经电泳得一条蛋白带,说明过氧化氢酶已经被提纯到电泳纯。光谱分析发现,甲醇处理后过氧化氢酶纯酶的吸收光谱和荧光发射光谱与未经处理的比较基本不变,而差示光谱出现明显的正峰和负峰。由动力学分析可知,在甲醇中,过氧化氢酶的Vmax和Km值均有不同程度提高 。     

The mechanism of the oxidation of d-amphetamine by rabbit liver oxygenase. Oxygen-18 studies

The oxidation of d-amphetamine by rabbit liver microsomes has been studied using oxygen-18 as the source of oxygen. Incorporation of heavy oxygen into the two major metabolites phenylacetone oxime and phenylacetone, was 93–95% and 25–31% respectively. These data are consistant with a mechanism in which the initial step is the hydroxylation of the substrate at the carbon atom α to the amino group. The carbinol amine which is formed by this reaction then serves as the key intermediate from which ketone and oxime are formed. Thus, oxime can form from carbinol amine in two step, (1) dehydration of carbinol amine and (2) oxygenation of the resulting imine. Phenylacetone can form by two pathways (1) loss of a molecule of ammonia from carbinol amine (incorporation of oxygen from molecular oxygen) and (2) hydrolysis of oxime (incorporation of oxygen from water). In the case of d-amphetamine the hydrolytic route appears to be the more important as suggested by Hucker, et al. (4, 5).     

A bis-[N-3-(1-hydroxy-1-methyl-ethyl)-benzyl)-cyclic urea as a HIV protease inhibitor

Synthetic efforts to overcome the metabolic oxidative degradation of the HIV protease inhibitory cyclic urea DMP323, a benzyl alcohol, have led to the discovery of a tertiary carbinol with superior affinity for the viral protease and more potent inhibitory activity against viral replication. Synthetic approaches to this new carbinol and comparative analysis of its pharmacology and pharmacokinetics are presented.     

蔓茎堇菜总黄酮提取工艺初探

分别采用热水浸提法、甲醇浸提法和乙醇浸提法来提取蔓茎堇菜样品的总黄酮,并以芦丁为标准品,用紫外分光光度法在265nm处测定各提取液的总黄酮含量。结果表明,甲醇浸提法和乙醇浸提法显著优于热水浸提法,而用70％乙醇为提取溶剂时效果最佳。研究结果可为利用蔓茎堇菜工业化生产黄酮类药用成分提供科学依据。     

Biotransformation of aromatic aldehydes bySaccharomyces cerevisiae: Investigation of reaction rates

Summary The rate of production ofl-phenylacetyl carbinol bySaccharomyces cerevisiae in reaction mixtures containing benzaldehyde with sucrose or pyruvate as cosubstrate was investigated in short 1 h incubations. The effect of yeast dose rate, sucrose and benzaldehyde concentration and pH on the rate of reaction was determined. Maximum biotransformation rates were obtained with concentrations of benzaldehyde, sucrose and yeast of 6 g, 40 g and 60 g/l, respectively. Negligible biotransformation rates were observed at a concentration of 8 g/l benzaldehyde. The reaction had a pH optimum of 4.0–4.5. Rates of bioconversion of benzaldehyde and selected substituted aromatic aldehydes using both sucrose and sodium pyruvate as cosubstrate were compared. The rate of aromatic alcohol production was much higher when sucrose was used rather than pyruvate.o-Tolualdehyde and 1-chlorobenzaldehyde were poor substrates for aromatic carbinol formation although the latter produced significant aromatic alcohol in sucrose-containing media. Yields of 2.74 and 3.80 g/l phenylacetyl carbinol were produced from sucrose and pyruvate, respectively, in a 1 h reaction period.     

Mosher ester analysis for the determination of absolute configuration of stereogenic (chiral) carbinol carbons

This protocol details the most commonly used nuclear magnetic resonance (NMR)-based method for deducing the configuration of otherwise unknown stereogenic, secondary carbinol (alcohol) centers (R1R2CHOH (or the analogous amines where OH is replaced by NH2)). This 'Mosher ester analysis' relies on the fact that the protons in diastereomeric alpha-methoxy-alpha-trifluoromethylphenylacetic acid (MTPA) esters (i.e., those derived from conjugation of the carbinol under interrogation with MTPA) display different arrays of chemical shifts (deltas) in their 1H NMR spectra. The protocol consists of the following: (i) preparation of each of the diastereomeric S- and R-MTPA esters and (ii) comparative (Delta delta(SR)) analysis of the 1H NMR spectral data of these two esters. By analyzing the sign of the difference in chemical shifts for a number of analogous pairs of protons (the set of Delta delta(SR) values) in the diastereomeric esters (or amides), the absolute configuration of the original carbinol (or amino) stereocenter can be reliably deduced. A typical Mosher ester analysis requires approximately 4-6 h of active effort over a 1- to 2-d period.     

黑龙江省西部乌拉尔甘草总黄酮含量的动态变化研究

采用超声提取和分光光度法对黑龙江省西部地区乌拉尔甘草不同部位的总黄酮含量进行测定,并对其季节变化进行了分析,研究结果表明:采用50%甲醇溶液提取,超声45 min对甘草总黄酮提取效果较好,适于进行大批量样品的提取测定.无论是野生甘草还是栽培甘草,在一个生长季中,叶的总黄酮含量最高,而地下部分含量则相对较低.在5～10月期间,叶的总黄酮含量逐渐下降,而地下部分总黄酮含量如根和根茎具有上升的趋势.甘草各部位总黄酮含量在不同生长季存在波动现象,尤其在具有运输功能的部位如复叶柄、地上茎表现更为明显.野生甘草不同部位的总黄酮含量的波动可能与有性繁殖有关,而栽培甘草的总黄酮含量的波动可能与无性生殖有关.     

Anilide tertiary carbinols: A new structural class of potent potassium channel openers

A new structural class of anilide tertiary carbinol potassium channel openers (PCOs) is described.     

Biotransformation of benzaldehyde by Saccharomyces cerevisiae: characterization of the fermentation and toxicity effects of substrates and products

Although higher initial rates of phenylacetyl carbinol formation were observed in fermentations containing a high starting benzaldehyde level, a massive reduction in yeast viability was observed resulting in early cessation of production formation. Pulse feeding to maintain lower benzaldehyde concentrations resulted in a lower initial reaction rate, but prolonged yeast viability and the biotransformation. This resulted in higher overall product tilers. As benzaldehyde concentration was increased, yeast growth rate was reduced (0.5 g/L), inhibited (1-2 g/L), or cell viability reduced (3 g/L). Benzaldehyde appeared to alter the cell permeability barrier to substrates and products. Reductions in yeast biomass levels and especially protein and lipid content were observed during the biotransformation. The effects of benzaldehyde and reaction products on yeast pyruvate decarboxylase and alcohol dehydrogenase stability were determined. Homogenized yeast cells produced similar phenylacetyl carbinol levels to whole yeast only if supplemented with thiamine pyrophosphate and magnesium.     

A new efficient synthesis of efaroxan

The key step of the synthesis of efaroxan was the dihydrobenzofuran ring formation involving an intramolecular cyclization of the tertiary alcohol intermediate with the fluoroaromatic moiety in basic medium. This carbinol was prepared according to two routes, either from reaction of a benzyl Grignard reagent with an alpha-ketoester, or from a Darzens condensation.     

Metabolism of monoterpenes: oxidation of isopiperitenol to isopiperitenone, and subsequent isomerization to piperitenone by soluble enzyme preparations from peppermint (Mentha piperita) leaves

Soluble enzyme extracts from peppermint leaves, when treated with polystyrene resin to remove endogenous monoterpenes and assayed with unlabeled substrates coupled with capillary gas-liquid chromatographic/mass spectrometric detection methods, were shown to oxidize isopiperitenol to isopiperitenone, and to isomerize isopiperitenone to piperitenone. The enzymes responsible for the monoterpenol dehydrogenation and the subsequent allylic isomerization were separated and partially purified by chromatography on Sephadex G-150, and were shown to have molecular weights of approximately 66,000 and 54,000, respectively. The general properties of the NAD-dependent dehydrogenase were examined, and specificity studies indicated that a double bond adjacent to the carbinol carbon was a required structural feature of the monoterpenol substrate. General properties of the isomerase were also determined, and it was demonstrated that the double bond migration catalyzed by this enzyme involved an intramolecular 1,3-hydrogen transfer. These enzymatic transformations represent two key steps in the metabolic pathway for the conversion of the initially formed cyclic olefin, (+/-)-limonene, to (-)-menthol and related monoterpenes characteristic of peppermint. Some stereochemical features of these reactions, and of the overall biogenetic scheme, are described.     

Characterization of the two major CYP450 metabolites of ozonide (1,2,4-trioxolane) OZ277

The antimalarial synthetic ozonide OZ277 (RBx11160) was hydroxylated by human liver microsomes at the distal bridgehead carbon atoms of the spiroadamantane substructure to form two carbinol metabolites devoid of antimalarial activity.     

Stereoselectivity in the oxidation of bufuralol,a chiral substrate,by human cytochrome P450s

Bufuralol (BF), a nonselective beta-adrenoceptor blocking agent, has a chiral center in its molecule, yielding the enantiomers 1'R-BF and 1'S-BF. beta-Adrenoceptor blocking potency is much higher in 1'S-BF than in 1'R-BF. One of the metabolic pathways of BF is 1"-hydroxylation of an ethyl group attached at the aromatic 7-position forming a carbinol metabolite (1"-hydroxybufuralol, 1"-OH-BF), and further oxidation (or dehydrogenation) produces a ketone metabolite (1-oxobufuralol, 1"-Oxo-BF). Both 1"-OH-BF and 1"-Oxo-BF are known to have beta-adrenoceptor blocking activities comparable to or higher than those of the parent drug. The 1"-hydroxylation introduces another chiral center into the BF molecule and four 1"-OH-BF diastereomers are formed from BF racemate in mammals, including humans, making elucidation of the metabolic profiles complicated. HPLC methods employing derivatization, reversed phase, or chiral columns have been developed to efficiently separate the four 1"-OH-BF diastereomers formed from BF enantiomers or racemate. Accumulated in vitro experimental results revealed that 1'R-BF is a much more preferential substrate than 1'S-BR for BF 1"-hydroxylation in human liver microsomes. Kinetic studies using recombinant human cytochrome P450 (CYP) enzymes indicate that CYP2D6 serves as a major BF 1"-hydroxylase and that CYP1A2 and CYP2C19 also contribute to BF 1"-hydroxylation in human livers. This mini-review summarizes the knowledge reported so far on the pharmacology of BF and its metabolites and the profiles of BF metabolism, especially focusing on the stereoselectivity in the oxidation of BF mainly in human livers and recombinant CYP enzymes.     

HPLC法测定黑龙江省不同地区刺五加紫丁香苷的含量

刺五加(Acanthopanax senticosus (Rupr. Et Maxim.)Harms)是五加科植物,是全球畅销的植物药之一,含有大量生物活性物质,其中所含有的刺五加苷类具有抗疲劳、抗衰老、抑制肿瘤生长等作用,紫丁香苷则是其中的主要活性成分之一。用HPLC法测定黑龙江省不同地区刺五加中紫丁香苷的含量,确定了刺五加紫丁香苷用超声波提取 1.5 h、60%甲醇作提取溶剂、提取温度为55℃的最佳提取条件组合。HPLC条件则是以检测波长为266 nm、流动相为甲醇/水（V/V）=28/72、流速为1 mL·min^-1、柱温为25℃。对黑龙江省不同地区类似生境条件下,同龄野生刺五加中紫丁香苷的含量进行比较,在所调查的7个地区中,以五营丰林国家自然保护区的生长在红松原始林下的刺五加紫丁香苷含量最高,五大连池雪洞风景区和牡丹江镜泊湖地区的刺五加紫丁香苷的含量最低。该研究结果对刺五加的采集和种植生境的选择具有积极的指导意义。