猴菇菌培养物中二个新吡喃酮化合物的分离与鉴定

从猴菇菌(Hericium erinaceus Bull pers)培养物中分得4个晶体和1个液态化合物。经鉴定,其中二个是新化合物,命名为猴菇菌素Ⅲ(Ⅲ)和Ⅳ(Ⅳ),其化学结构分别定为6-甲基-2,5-二羟甲基-γ-吡喃酮(Ⅲ)和2-羟甲基-5-(1′-羟基)-乙基-γ-吡喃酮(Ⅳ)。其余三个晶体的化学结构分别为4-氯-3,5-二甲氧基苯甲酸-O-阿拉伯糖醇酯(Ⅰ),4-氯-3,5-二甲氧基苯甲酸甲酯(Ⅲ),4-氯-3,5-二甲氧基苯甲酸(Ⅴ)。     

Metabolites of microorganisms. 247. Phenazines from Streptomyces antibioticus, strain Tü 2706

From a strain of Streptomyces antibioticus seven yellow phenazines were isolated. The antibacterially most active antibiotic was identified as (-)-saphenamycin, a second one with compound DC-86-Y (saphenic acid). Three compounds were new: Saphenic acid methyl ether, 6-acetylphenazine-1-carboxylic acid and an inseparable mixture of fatty acid esters of saphenic acid. Two simple phenazines were phenazine-1-carboxylic acid (tubermycin B) and unsubstituted phenazine, which was isolated for the first time from a microorganism.     

Multichannel Liquid Chromatography–Tandem Mass Spectrometry Cocktail Method for Comprehensive Substrate Characterization of Multidrug Resistance-Associated Protein 4 Transporter

Purpose To develop a comprehensive substrate-screening method for the ATP-binding cassette (ABC) transporter, and identify new substrates for multidrug resistance-associated protein 4 (MRP4/ABCC4). Methods Human MRP4-expressing membrane vesicles were incubated with a mixture of 50 compounds, including methotrexate, a known MRP4 substrate. The amounts transported were simultaneously determined by liquid chromatography–tandem mass spectrometry. Results From 49 compounds, 12 were identified as substrate candidates for MRP4 in the first screening. The second screening was performed involving the uptake of mixture using single quadrupole multichannel mode, and the third screening was performed involving the uptake of individual compounds using multiple reaction monitoring multichannel mode. As a result, eight substrate candidates were additionally identified. Subsequently, in the fourth step, osmotic pressure-dependent transport was demonstrated for 18 compounds (cefmetazole, piperacillin, rebamipide, tetracycline, ampicillin, benzylpenicillin, bumetanide, cephalosporin C, enalapril, pipemidic acid, furosemide, ceftazidime, pravastatin, hydrochlorothiazide, sulbactam, baclofen, bezafibrate and alacepril) among the 20 substrate candidates, thereby confirming them as MRP4 substrates. By contrast, the uptakes of meloxicam and nateglinide did not depend on osmolarity, indicating that these compounds were not substrates, but bound to MRP4. Conclusions The new comprehensive substrate-screening method for ABC transporters allowed the identification of 18 new substrates for MRP4.     

猫爪草化学成分的研究

猫爪草化学成分的研究郭学敏，周卓轮，洪永福（上海第二军医大学药学院中西药研究室２００４３３）猫爪草为毛莨科植物小毛莨（ＲａｎｎｕｃｕｌｕｓｔｅｒｎａｔｕｓＴｈｕｎｂ．）的块根，收载于１９９０年版中国药典一部．猫爪草在中医临床上用于治疗咽喉炎、肺结核、...     

Design and synthesis of some new thiazolo [3,2-b]-1,2,4-triazole-5(6H)-ones substituted with flurbiprofen as anti-inflammatory and analgesic agents

In the course of our ongoing studies, a series of thiazolo[3,2-b]-1,2,4-triazole-5(6H)-ones substituted with flurbiprofen (CAS 5104-49-4) has been prepared. The compounds were synthesized by the cyclization of the 3-[(2-fluoro-4-biphenyl)ethyl]-5-mercapto-1,2,4-triazole (3) with chloroacetic acid and relevant benzaldehydes in the presence of acetic acid, acetic anhydride and anhydrous sodium acetate in one step. The product of this one-pot synthesis that precipitated on cooling of the reaction mixture was identified undoubtedly by X-ray crystallographic analysis as thiazolo[3,2-b]-1,2,4-triazole. In-vivo anti-inflammatory and analgesic activities of the compounds were assessed by carrageenan-induced hind paw edema and p-benzoquinone-induced abdominal constriction tests in mice, respectively. In addition, the ulcerogenic risks were evaluated. It is worthy of saying that the compounds which maintained analgesic/antiinflammatory activity of the starting compound were found to be safer with regard to gastric lesion risks at 100 mg/kg oral dose when compared with flurbiprofen. Among the synthesized compounds 3d showed the highest analgesic and antiinflammatory activity without inducing any gastric lesion and deserves further attention in order to develop new lead drug candidates.     

Triterpenes and sterols in the flowers and leaves of Prunus spinosa L. (Rosaceae)

From the flowers of Prunus spinosa L. a mixture of alpha- and beta-amyrine, a mixture of ursolic and oleanolic acids, ursolic acid, a mixture of beta-sitosterol, gamma-sitosterol and stigmasterol and beta-sitosterol 3-O-beta-D-glucopyranoside were isolated. Their structure was determined by spectroscopic methods (GC/MS, IR, 1H and 13C NMR). Chromatographic methods (GC, HPLC, TLC) were employed to determine the presence of the listed compounds in the leaves. Also, the content of beta-sitosterol, ursolic and oleanolic acids in the leaves and flowers of blackthorn was determined by GC/MSD method.     

Isolation from Cussonia barteri of 1'-O-chlorogenoylchlorogenic acid and 1'-O-chlorogenoylneochlorogenic acid, a new type of quinic acid esters.

1'-O-Chlorogenoylchlorogenic acid and 1'-O-chlorogenoylneochlorogenic acid, a new type of quinic acid esters, have been isolated, in addition to six known quinic acid esters, rutin, and a mixture of saponins, from the methanol extract of Cussonia barteri Seemann (Araliaceae) leaves collected in Cameroon. Structure determination was achieved by NMR, mass, IR, and UV spectroscopy. All compounds were tested for inhibitory activity on 5-lipoxygenase and cyclooxygenase-1, for antimicrobial activity against Bacillus subtilis, Pseudomonas fluorescens, and Cladosporium cucumerinum, and for haemolytic activity.     

人参皂苷元25-OH-PPD的修饰及结构研究

目的设计合成3β,12β,20,25-四羟基-达玛烷(25-OH-PPD)的衍生物,并确定其结构。方法以25-OH-PPD为原料,以DCC、DMAP为催化剂,在室温下与饱和脂肪酸进行酯化反应得到一系列不同位置取代的25-OH-PPD酯类衍生物,经IR、13C-NMR、HR-MS确证其结构。结果与结论设计并合成了8个未见文献报道的人参皂苷元25-OH-PPD衍生物。该工艺条件温和,易于操作,为原人参二醇型衍生物的构效关系研究奠定基础。     

The novel formulation design of O/W microemulsion for improving the gastrointestinal absorption of poorly water soluble compounds

The design of the novel O/W microemulsion formulation, which enhances the oral bioavailability by raising the solubility of poorly water soluble compounds was examined. Using medium chain fatty acid triglyceride (MCT), diglyceryl monooleate (DGMO-C), polyoxyethylene hydrogenated castor oil 40 (HCO-40), ethanol and PBS (pH 6.8) as an oil phase, a lipophilic surfactant, a hydrophilic surfactant, a solubilizer and an aqueous phase, at the mixture ratio of 5%/1%/9%/5%/80% (w/w), respectively, the O/W microemulsion with an average particle diameter of 20 nm or less was prepared. Moreover, for nine kinds of poorly water soluble compounds, such as Ibuprofen, Ketoprofen, Tamoxifen, Testosterone, Tolbutamide and other new compounds, the solubility to water was increased from 60 to 20,000 times by this O/W microemulsion formulation. The AUCs in plasma concentration of Ibuprofen and a new compound, ER-1039, following single oral administration of these compounds as the O/W microemulsion to fasted rats were equivalent to that of solution administration or increased by nine and two times that of suspension administration, respectively. Accordingly, this novel O/W microemulsion is a useful formulation, which enhances the oral bioavailability by raising the solubility of poorly water soluble compounds.     

N-deacetyl-N-aminoacylthiocolchicine derivatives: synthesis and biological evaluation on MDR-positive and MDR-negative human cancer cell lines

A new series of N-deacetyl-N-(N-trifluoroacetylaminoacyl)thiocolchicine derivatives 9-15 have been synthesized starting from the corresponding N-deacetylthiocolchicine (3) and the N-trifluoroacetylamino acids 5-8 which were used as a racemic mixture. The trifluoroacetyl protecting group has been removed easily, giving the corresponding N-deacetyl-N-aminoacylthiocolchicines 16-22. Optical pure compounds 9-22 were isolated from the diastereoisomeric mixture or were prepared starting from compound 3 and an optical pure amino acid derivative; the configuration of each compound was assigned unequivocally. The diastereoisomeric couples of amino acids synthesized were tested, and their antiproliferative activity on MDR-positive and MDR-negative human cancer cell lines was evaluated.     

Comparative study of antithrombotic and antiaggregatory activity of acetylsalicylic acid, ticlopidine and a new noncarboxylic acid antiinflammatory pyrazine derivative HF90

The action of acetylsalicylic acid, ticlopidine and a new pyrazine derivative HF90 selected in preliminary screenings (11, 18, 19) was studied by using the mouse antithrombotic assay according to DiMinno and Silver (22) and in vitro blood platelet aggregation method according to Born (23). Acute pulmonary thromboembolism was induced by injection of a mixture of collagen and epinephrine into the mouse tail vein. The effect of HF90, an acidic pyrazine derivative possessing active methylene moiety, administered at doses of 50 and 100 mg/kg, was compared to the action of the well established antithrombotic agents: ticlopidine (100 mg/kg) and acetylsalicylic acid (20 mg/kg). The compounds were administered i.p. in single doses 1 h and 24 h before the thrombotic challenge or once a day per three consecutive days before the thrombotic challenge. Ticlopidine appeared to provide the better protection against microembolism than acetylsalicylic acid although its effect has not manifested itself immediately after administration. The pyrazine derivative examined has a lower but significant antithrombotic activity. The chemical class of pyrazine derivatives with active methylene moiety (the so called pyrazine CH/NH-acids) (16) provides a new original antiinflammatory pharmacophore and HF90 may serve as the "lead compound" in the search for new agents of pharmacological interest.     

A new simplified assay for the quantitation of theophylline and its major metabolites in urine by high-performance liquid chromatography

A high-performance liquid chromatographic technique is presented for the simultaneous determination of theophylline, 3-methylxanthine, 1-methyluric acid, 1,3-dimethyluric acid, and caffeine in urine using beta-hydroxyethyl theophylline (BHET) as an internal standard. Following centrifugation of the sample, 2 mL of urine was placed onto a Sep-Pak C18 cartridge (Waters Associates, Milford, MA, USA) that was prepared with 5 mL of methanol followed by 10 mL of distilled water. The compounds were eluted with 2.5 mL of a methanol:water (50:50) mixture. An aliquot of the filtered urine sample (100 microL) was added to 40 microL of an internal standard solution (BHET; 100 micrograms/mL). Distilled water (60 microL) was added to yield a volume of 200 microL. The mixture was vortexed and centrifuged, and 100 microL was injected onto a C8 column maintained at 55 degrees C. Inter- and intraday variability of the assay for all compounds was less than 5.7%. Sensitivity ranged from 0.10 microgram/mL for caffeine to 0.68 microgram/mL for 1-methyluric acid. Drugs commonly coadministered with theophylline did not interfere with the assay.     

Diterpenes isolated from Croton zambesicus inhibit KCl-induced contraction

A mixture of two new diterpenes was isolated from a dichloromethane extract of Croton zambesicus: ent-18-hydroxytrachyloban-3beta-ol (1) and ent-18-hydroxyisopimara-7,15-diene-3beta-ol (2). The two compounds crystallised together and were separated after derivatisation of the pimarane derivative with osmium tetroxide. The structure of 1 was elucidated by 1D- and 2D-NMR analysis and by X-ray diffraction of a crystal containing both compounds while 2 was only identified by crystallographic data. As this plant is widely used in African folk medicine against hypertension, we have analysed the vasorelaxant activity of the isolated molecules. The mixture of the two compounds inhibited the KCl-induced contraction of male Wistar rat aorta (IC (50) = 1 microg/mL), while the purified trachylobane (compound 1) and the hydroxylated pimarane showed a lower activity than the mixture.     

The novel formulation design of self-emulsifying drug delivery systems (SEDDS) type O/W microemulsion I: enhancing effects on oral bioavailability of poorly water soluble compounds in rats and beagle dogs

We examined the design of the versatile novel self-emulsifying drug delivery systems (SEDDS) type O/W microemulsion formulation which enhances the oral bioavailability by raising the solubility of poorly water soluble compounds. Namely, seven kinds of poorly water soluble compounds such as disopyramide, ibuprofen, ketoprofen, tolbutamide, and other new compounds, as the model compounds were used to compare the plasma concentration profile of the compound following single oral administration of each compound to rats and beagle dogs as a solution, an oily solution, a suspension (or a powder), an O/W microemulsion, and a SEDDS type O/W microemulsion. And the enhancing effect of the SEDDS type O/W microemulsion on the gastrointestinal absorption of these compounds was evaluated. In the components of the SEDDS type O/W microemulsion, medium chain fatty acid triglyceride (MCT), diglyceryl monooleate (DGMO-C), polyoxyethylene hydrogenated castor oil 40 (HCO-40), and ethanol were used as an oil, a lipophilic surfactant, a hydrophilic surfactant, and a solubilizer, at the mixture ratio of 25/5/45/25 (w/w%), respectively. Thereby, to six kinds of the model compounds except disopyramide, the solubility was from 340 to 98,000 times that in water, and the AUCs in plasma concentration of the compound were equivalent to that of solution or O/W microemulsion administration, or was increased by 1.5 to 78 times that of suspension administration. Accordingly, this novel SEDDS type O/W microemulsion is the versatile, useful formulation which enhances the oral bioavailability by raising the solubility of poorly water soluble compounds.     

Comparison of the effect of Aminopielik D pesticide and its active components on human erythrocytes

In the present work, the effect of Aminopielik D [417.5g/l of dimethylamino salts of 2,4-dichlorophenoxyacetic acid (2,4-D) and 32.5g/l of 3,6-dichloro-2-metoxybenzoic acid (Dicamba)] and its active components (used separately and in mixture) on human erythrocytes was examined. The parameters studied were: lipid peroxidation, metHb formation and catalase activity. Aminopielik D used at doses of 100-1000ppm was found to increase lipid peroxidation, decrease of catalase activity and oxidation of haemoglobin. 2,4-D and Dicamba are present in Aminopielik D in the dimethylamino form; their sodium salts in solution (separately and as a mixture) did not cause such strong effects. A synergistic action of 2,4-D and Dicamba was excluded as the individual compounds caused the same effects as their mixture. Aminopielik D provoked slightly higher changes in the lipid peroxidation and catalase activity than its active components alone and in mixture, which was probably a result of the properties of the additives and interaction of tested systems with the dimethylamino group.     

Determination of antileukemic diimidazolinyl compounds in plasma of experimental animals by high-performance liquid chromatography

A method is reported for the determination of the fluorescent, antileukemic diimidazolinyl compounds 261/96(1). 253/152(2) and 272/131(3) in plasma. HPLC is performed on a RP-2 10 microns column with a mobile phase of methanol/water (1:1, v/v), to which 0.005 mol/l octanesulfonic acid sodium and 0.003 mol/l dimethyloctylamine are added. Samples are prepared by precipitation of plasma proteins with a mixture of methanol-70% perchloric acid. The assay is linear up to 750 ng/ml for all compounds with limits of determination of 4 ng/ml, 2 ng/ml and 0.5 ng/ml for compounds 1, 2 and 3, respectively. Coefficients of variation are below 10 percent at all concentrations studied.     

Antisickling activity of hydroxybenzoic acids in Cajanus cajan.

The amounts of phenylalanine and hydroxybenzoic acid in a Cajanus cajan methanolic extract were estimated. Results showed that the amount of phenylalanine and hydroxybenzoic acid per gram weight of bean was 4.92 mg +/- 0.13 mg and 21.0 mg +/- 3.0 micrograms, respectively. Sickling inhibition was observed to be efficient with the extract which contains a mixture of phenylalanine (0.69 mg/ml) and p-hydroxybenzoic acid (10.5 micrograms/ml), equivalent to those found in bean extract. The additive antisickling effect of both compounds can be therapeutically exploited for the treatment of sickle cell anemia.     

The single substance and mixture toxicity of quinolones to the bioluminescent bacterium Vibrio fischeri

Quinolones are one of the most important group of synthetic antibiotics used in aquaculture. We studied the single substance and mixture toxicity of ten quinolones using a long term bioluminescence inhibition assay with the marine bacterium Vibrio fischeri as the test organism. All tested quinolones are highly toxic to the test organism with EC50 values ranging from 14 μg/l for ofloxacin to 1020 μg/l for pipemidic acid. Adapting the approach outlined in EEC directive 93/21/EEC to these results, all but one of the ten quinolones belong to the group classified as being 'very toxic to aquatic organisms' (EC50 below 1 mg/l). On the basis of the concentration-response relationships of the single compounds, the mixture toxicity of the ten compounds was estimated by the concepts of concentration addition and independent action. Complete concentration-response relationships were experimentally recorded for the quinolone mixture in three different mixture ratios, based on the relative toxicity of the components (EC50, EC1 and NOEC). The results show that the mixture toxicity of the quinolones is best predictable by concentration addition, whereas independent action underestimates the toxicity of the mixture. As the quinolones have an identical specific mechanism of action (the inhibition of bacterial gyrases), these results are in agreement with the pharmacological assumptions that form the basis of the concept of concentration addition. It is therefore concluded, that concentration addition can be useful for hazard assessment procedures of mixtures of similarly acting compounds. One important implication of this concept is that even mixture components that are present only at their individual no observed effect concentrations (NOECs) contribute to the overall toxicity of the mixture. Under these conditions more than 99% effect of the quinolone mixture are observed. This result emphasises the unsuitability of NOECs as an approximation of a 'safe' concentration.     

Antitussive,expectorant, and anti-inflammatory activities of four caffeoylquinic acids isolated from Tussilago farfara

Context: The flower bud of Tussilago farfara L. (Compositae) (FTF) is one of the traditional Chinese medicinal herbs used to treat cough, phlegm, bronchitic, and asthmatic conditions.

Objective: The objective of this study is to isolate four caffeoylquinic acids from the ethyl acetate extract (EtE) of FTF and to evaluate their antitussive, expectorant, and anti-inflammatory activities.

Materials and methods: The structures of compounds 1–4 isolated from EtE were determined by spectral analysis. Mice were orally treated with these compounds and their mixture (in a ratio of 5:28:41:26 as in EtE) at doses of 10 and 20?mg/kg once daily for 3 d. The antitussive and expectorant activities were evaluated separately with the ammonia liquor-induced model and the phenol red secretion model. The anti-inflammation activity was evaluated using leukocyte count in the bronchoalveolar lavage fluid after ammonia liquor-induced acute airway inflammation.

Results: The four compounds were identified as chlorogenic acid (1), 3,5-dicaffeoylquinic acid (2), 3,4-dicaffeoylquinic acid (3), and 4,5-dicaffeoylquinic acid (4). All compounds, especially compound 4 (58.0% inhibition in cough frequency), showed a significant antitussive effect. However, the mixture was the most effective to inhibit the cough frequency by 61.7%. All compounds also showed a significant expectorant effect, while compound 2 was the most potent to enhance the phenol red secretion by 35.7%. All compounds significantly alleviated inflammation, but compound 4 showed the strongest effect to inhibit the leukocytosis by 49.7%.

Discussion and conclusion: The caffeoylquinic acids and their mixture, exhibiting significant antitussive, expectorant, and anti-inflammatory effects, could be considered as the main effective ingredients of FTF, and they may act in a collective and synergistic way.     

Microdontin A and B: Two New Aloin Derivatives from Aloe microdonta

The leaf juice of ALOE MICRODONTA Chiov. is used in Somali traditional medicine as a remedy for jaundice and for topical treatment of skin diseases. Mucilage was precipitated from the fresh juice by addition of ethanol and the supernatant chromatographed on Amberlite XAD-2 to yield a fraction containing anthraquinone derivatives. Flash chromatography on silica gel followed by preparative TLC yielded aloin A and a new compound. Spectral data (mass, (1)H- and (13)C-NMR) identified this compound as a mixture of aloin A and B where the glucose of both compounds is esterified with 4-hydroxycinnamic acid at the 6'-position. The two compounds were denoted as microdontin A and B, respectively.