Optimization and validation of a chromatographic method for the simultaneous quantification of six bioactive compounds in Rhizoma et Radix Polygoni Cuspidati

A reverse-phase HPLC method was developed for simultaneous quantification of six bioactive compounds in Rhizoma et Radix Polygoni Cuspidati. These compounds--polydatin (1), resveratrol (2), rhein (3), emodin (4), chrysophanol (5) and physcion (6)--were analysed from 24 authentic samples of the herb using UV HPLC. Based on the UV absorption characteristics of the six compounds, absorption wavelengths of 306 nm were chosen to quantify compounds 1 and 2, and 290 nm for compounds 3-6. A reliable and reproducible quantitative HPLC method for analysing authentic samples of Rhizoma et Radix Polygoni Cuspidati from different cultivation regions was developed. The results showed that the concentration of compound 1 in samples from Sichuan was almost 2 fold higher than that of samples acquired in Guangxi. Furthermore, compounds 3 and 5 were not found in all the samples tested. Thus, instead of using polydatin (1) and emodin (4) as markers for quality assessment, as in conventional practice, these findings show that compounds 2 and 6 are more suited to act as marker compounds for a more specific assessment of the quality of this herb.     

Metabolism in Rats of Several Carboxylic Acid Derivatives Containing the 3,4-Methylenedioxyphenyl Group

Abstract: The metabolism of the 3,4-methylenedioxy derivatives of mandelic acid (1), phenylacetic acid (2), benzoic acid (3), 3-phenylpropionic acid (4) and cinnamic acid (5) was studied in rats. Following intragastric dosage (1 mmol/kg) the compounds and their metabolites were excreted in the urine within 24 hrs. Recoveries of roughly 85% were obtained. Except for compound (1) which was excreted to a large extent unchanged, glycine conjugates were the major urinary metabolites. Compound (2) formed 3,4-methylenedioxyphenylacetylglycine whereas compounds (3), (4) and (5) were converted to 3,4-methylenedioxybenzoylglycine. No evidence was found with any of the compounds for demethylenation and subsequent excretion of catecholic metabolites.     

Synthesis and antimicrobial activity of 3-alkoxyjatrorrhizine derivatives

The compounds 3-ethoxy- ( 2), 3-butoxy- ( 3), 3-hexyloxy- ( 4), 3-octyloxy- ( 5), 3-decyloxy- ( 6) and 3-dodecyloxyjatrorrhizine chlorides ( 7) were synthesized and tested for their antimicrobial activity in vitro to evaluate structure-activity relationships. Substitution of the H with alkyl groups at C-3-OH led to significant changes in the antimicrobial activity. The antimicrobial activity of the substituted derivatives was 32 - 1000 times higher than that of jatorrhizine ( 1), which increased as the aliphatic chain was elongated and then decreased slightly when the alkyl chain exceeded eight carbon atoms. 3-Octyloxyjatrorrhizine ( 5) displayed the highest antimicrobial activity of all compounds. The LD (50) values of compounds 1 - 7 were more than 6000 mg/kg body weight, showing a low toxicity. The toxicities of compounds 2 - 7 were slightly lower than that of ( 1).     

Chemical constituents from the Moutan Cortex charcoal and their potential coagulation activities

The chemical constituents of Cortex Moutan charcoal were studied in depth, and their coagulation activity was screened. The chemical constituents were isolated by polyamide, silica gel and Sephadex G-Sepharose chromatography purification,and their structures were identified by NMR spectroscopy and other analyses. A total of 10 compounds were obtained and identified as follows: 3-(2-furan)-1-(2-hydroxy-4-methoxy-phenyl)-2-propylene ketone (1), 2-(2,5-hydroxy-4-methyl phenyl) propionate ethyl ester (2), methyl tetradec-5-enoate (3), 1,4-diethylcyclohexane (4), catechol (5), methyl-4-hydroxybenzoate (6), 3-hydroxy-2-methyl-4-pyrone (7), 3,8-dihydroxy-2-metyl-chromone (8), 3β-hydroxy-olean-12-en (9), 1-monolinolein (10). Among them, compounds 1 and 2 were new natural products, and 3–10 were isolated from the Cortex Moutan charcoal for the first time. The potential coagulation activities of compounds were evaluated, and compounds 2, 9 and 10had strong hemostatic effects. While compounds 1 and 8 could significantly activate blood circulation.     

Alkyl derivatives of isoquinoline. III. Synthesis and pharmacologic activity of dialkylaminoalkyl amides of 1-chloro- and 1-methoxy-3-carboxy-4-methylisoquinoline and of 3-carboxy-2,4-dimethyl-1,2-dihydro-1-oxoisoquinoline

The synthesis of a series of dialkylaminoalkylic amides of 1-chloro-3-carboxy-4-methylisoquinoline (VIII), of 1-methoxy-3-carboxy-4-methylisoquinoline (VI) and of 3-carboxy-2,4-dimethyl-1-oxoisoquinoline (XIII) is described. In addition a series of 1-amino-substituted 3-carboxymethyl-4-methylisoquinolines (II) was synthesized. The pharmacological activity of some of these compounds was studied. The compounds (XIIIa), (VIa) and (VIIIa) showed a clear local anaesthetic activity, a little lower than that of lidocaine; the same compounds also showed fairly good antispasmodic properties.     

海洋真菌Penicillium sclerotiorum GZU-XW03-2来源的杂萜类成分研究

摘要：目的 为获得结构多样、活性显著的次级代谢产物,对一株海洋来源真菌Penicillium sclerotiorum GZU-XW03-2的化学成分进行研究。方法 菌种经过活化后,经大米固体培养基大规模发酵后的菌丝体用甲醇浸提,旋干后,用水混悬,接着用乙酸乙酯萃取得粗提物。乙酸乙酯萃取物经硅胶色谱、凝胶色谱、反相色谱及高效液相色谱等柱色谱分离,对获得的单体成分采用核磁共振（NMR）、单晶衍射、电子圆二色谱（ECD）、高分辨质谱（HRESIMS）等手段进行鉴定,结合相关文献数据从而确定化合物结构。结果 从乙酸乙酯部位共分离获得六个杂萜化合物,依次为preaustinoid A6 (1)、1-oxymethyl-hydropreaustinoid A1 (2)、preaustinoid A1 (3)、miniolutelide A (4)、miniolutelide B (5)、chrysogenolide C (6)。结论 以上六个化合物均属于由DMOA途径衍生合成的杂萜类化合物,并且化合物1-6均从Penicillium sclerotiorum真菌中首次分离获得,其中A环开环的化合物1,2在天然产物中并不常见,此前共报道此类型化合物仅4个,化合物4的单晶数据为首次报道。     

Phenolic compounds from Baseonema acuminatum leaves: isolation and antimicrobial activity

Three new phenolic compounds, 1-galloyl-beta-D-glucopyranosyl-(1-->4)-beta-D-galactopyranoside (1), 2-methoxy-5-(1 '2 3'-trihydroxypropyl)-phenyl- 1-0-(6"-galloyl)-beta-D-glucopyranoside (2),and 2-methoxy-5-hydroxymethyl-phenyl-1-O-(6"-galloyl)-beta-D-glucopyranoside (3), together with the known compounds benzyl 6'-O-galloyl-beta-D-glucopyranoside (4), 1,6-di-O-galloyl-beta-D-glucopyranose (5), myrciaphenone B (6), kaempferol 3-0-(6"-galloyl)-beta-D-glucopyranoside (7), quercetin 3-0-(6"-galloyl)-beta-D-glucopyranoside (8), vomifoliol 9-O-beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranoside, 2,3-dihydrobenzofuran-2-(4'-hydroxy-3'-methoxyphenyl)-3-alpha-L-rhamnopyranosyloxymethyl-7-methoxy-5-propanol, and benzyl-O-alpha-L-rhamnopyranosyl-(1-->6)-Beta-D-glucopyranoside were isolated from the leaves of Baseonema acuminatum P. Choux (Asclepiadaceae). Their structures were determined by 1D- and 2D-NMR spectroscopy and by ESI-MS analysis. The antimicrobial activity of all compounds was evaluated in vitro against bacteria (Staphylococcus aureus two strains, Bacillus cereus, Bacillus subtilis, Escherichia coli, Salmonella thyphimurium) and three strains of Candida albicans. The new compounds 2 and 3, together with the known compound 4, showed antifungal activity against two clinically isolated Candida albicans strains and against C. albicans ATCC 2091; MIC values were in the range of 25-100 microg/mL. Compound 5 was active against the two clinically isolated strains of C. albicans with MICs of 12.5 microg/mL and 25 microg/mL. Compounds 1, 6, 7, and 8 inhibited only one strain of C albicans at the maximum concentration used. None of the phenolic compounds tested was active against the bacteria studied.     

Pharmacological Studies on Linderae umbellatae Ramus, IV*. Effects of Condensed Tannin Related Compounds on Peptic Activity and Stress-Induced Gastric Lesions in Mice

Nine condensed tannin related compounds isolated from LINDERAE UMBELLATAE Ramus, (+)-catechin ( 1), (-)-epicatechin ( 2), proanthocyanidin B-1 ( 3), B-2 ( 4), B-4 ( 5), trimer A ( 6), trimer B ( 7), cinnamtannin B (1) ( 8) and cinnamtannin D (1) ( 9), were tested for their effects on peptic activity and stress-induced gastric lesions in mice. Peptic activity of rat gastric juice was suppressed by 6 and 7 IN VITRO, and slightly suppressed by 8 and 9. In pylorus-ligated mice, these compounds, except for 4, administered orally exhibited anti-peptic activity. Pretreatments of 4,6 and 7 orally protected mice against stress-induced gastric lesions. Effects of these compounds on digestive systems were dìscussed.     

Cellular effects of some metabolic oxidation products pertinent to 4-ethoxyaniline.

The toxicity of some metabolic products pertinent to 4-ethoxyaniline in isolated hepatocytes were investigated. The compounds investigated were 4-ethoxynitrosobenzene (1), 4-ethoxy-4'-nitrosodiphenylamine (2), 3,6-bis(4-ethoxy-phenylimino)-4-ethoxy-1,4-cyclohexadienylamine (3), 4-(4-ethoxyphenylimino)-2,3-dimethyl-2,5-cyclohexadiene-1-one (4) and 4-(4-ethoxyphenylimino)-2,6-dimethyl-2,5-cyclohexadiene-1-one (5). Of these, 1, 2 and 3 are oxidation products of 4-ethoxyaniline. Compounds 4 and 5 are dimethyl analogues of previously investigated oxidation product 4-(4-ethoxyphenylimino(-2,5-cyclohexadiene-1-one (NEPBQI). Among the investigated compounds, 1 and 2 were the most toxic towards isolated hepatocytes. In hepatocytes treated with compounds 1, 2 and 4, loss of cell viability was also accompanied by surface bleb formation. All compounds except 3 reacted with GSH resulting in depletion of cellular GSH. No formation of GSSG was observed, however. Thus, the GSH depletion was apparently due to conjugate formation rather than oxidation. No superoxide dismutase inhibitable reduction of acetylated cytochrome c was observed, thus none of the compounds undergoes measurable redox cycling.     

溪桫化学成分研究

目的:研究楝科溪桫属植物溪桫[Chisocheton cumingianus subsp.balansae(C.Candolle)Mabber-ley]的小枝及茎皮化学成分。方法:利用正相、反相硅胶柱色谱、Sephadex LH-20凝胶柱色谱、制备HPLC等方法分离化合物,运用MS和NMR等波谱方法解析鉴定化合物结构。结果:从其乙醇提取物之三氯甲烷萃取部位中分离鉴定出10个化合物,分别为4个三萜类化合物:odoratone(1),grandifoliolenone(2),24-表-匹西狄醇A(24-epi-piscidinol A,3),chisiamol F(4);3个倍半萜类化合物:1β,6α-二羟基-4(15)-桉叶烯(1β,6α-dihydroxy-4(15)-eudesmene,5),1β,8α-二羟基-4(15)-桉叶烯(1β,8α-dihydroxy-4(15)-eudesmene,6),环氧泽泻烯(alismoxide,7);3个蒽醌类化合物:大黄酚(8),大黄素(9),大黄素甲醚(10)。结论:10个化合物均首次从该植物中分离得到。     

Anti-herpes simplex virus type-1 flavonoids and a new flavanone from the root of Limonium sinense

From the root of Limonium sinense (Girard) Ktze a new (2R,3S)-3,5,7,4'-tetrahydroxy-3',5'-dimethoxyflavanone was isolated and named isodihydrosyringetin (3), together with nine other known compounds, (-)-epigallocatechin 3-O-gallate (1), samarangenin B (2), myricetin (4), myricetin 3-O-alpha-rhamnopyranoside (5), quercetin 3-O-alpha-rhamnopyranoside (6), (-)-epigallocatechin (7), gallic acid (8), N-trans-caffeoyltyramine (9), and N-trans-feruloyltyramine (10). All of them were examined for their inhibitory effects on herpes simplex virus type-1 (HSV-1) replication in Vero cells. Both compounds 1 and 2 exhibited potent inhibitory activities in HSV-1 replication. Comparison of the IC50 values indicated that compounds 1 and 2 had higher inhibitory activities than the positive control acyclovir (38.6 +/- 2.6 vs. 55.4 +/- 5.3 microM, P < 0.001; 11.4 +/- 0.9 vs. 55.4 +/- 5.3 microM, P < 0.0005). Cytotoxicity was unlikely involved because no cell deaths were observable in the Vero cells following 5 day treatments with compound 1 or 2.     

南极海绵共附生真菌Talaromyces sp. HDN1820200次级代谢产物研究

目的对南极海绵共附生来源真菌Talaromyces sp.HDN1820200的次级代谢产物进行研究。方法对菌株发酵液粗提物利用柱层析(VLC,ODS)、凝胶分子筛(LH-20)以及半制备高效液相(MPLC)等方法进行次级代谢产物的分离纯化;利用现代波谱学方法(核磁共振NMR、圆二色谱ECD、红外光谱IR等)对化合物进行结构鉴定。结果分离鉴定了8个单体化合物:(2E,4E,6E)-4-methyldodeca-2,4,6-trienoic acid(1)、(2E,4Z,6E)-4-methyldodeca-2,4,6-trienoic acid(2)、Herbaridine B(3)、Kasanosin C(4)、Citrinolactone A(5)、Citreorosein(6)、1,5,8-trihydroxy-3-(hydroxymethyl)anthracene-9,10-dione(7)和Rugulosin B(8)。结论从南极海绵来源真菌Talaromyces sp.HDN1820200中分离得到了化合物1~8,其中化合物1和2为新化合物,属于脂肪酸类化合物,无明显细胞毒活性。     

New 8-substituted xanthiene derivatives as potent bronchodilators

The synthesis and structure determination of 8-aryl /alkyl aryl 1, 3-dimethyl-3, 7-dihydropurin-2, 6-dione derivatives (1-13), was carried out in this study. Bronchodilator activity is investigated using isolated guinea-pig tracheal strips, pre-contracted by acetylcholine and histamine. Spasmolytic activity of the compounds was compared to theophylline. Synthesized compounds (1-13) did not inhibit the acetylcholine-induced pre-contractions except compound (8) at 10(-5) M concentration. In contrast, some of the compounds, especially (7), (11), (12) at 10(-5) M and (3), (4), (9) and (11) in 10(-4) M displayed inhibitory activity on the tracheal strips pre-contracted by histamine. The potency of the compounds at human adenosine receptors was evaluated using radioligand binding assay and a cyclic AMP functional assay in CHO cells expressing these receptors. Compound (11) displayed the greatest activity against radioligand binding of specific agonists to A2A and A2B receptors. The compounds were relatively selective for both A2A and A2B compared with A1 and A3 receptors. All compounds were also tested for the inhibition of NECA-induced cAMP accumulation mediated by the A2B adenosine receptor and compound (11) was found to be the most effective. Our results showed that these compounds are acting as selective adenosine antagonists, especially for adenosine A2B receptors, and are promising as potent anti-inflammatory agents rather than bronchodilator for the treatment of asthma.     

Effect of Baccharis dracunculifolia D.C. (Asteraceae) extracts and its isolated compounds on macrophage activation

Baccharis dracunculifolia D.C. (Asteraceae), a shrub which grows wild in Brazil, is the main botanical source of Brazilian green propolis. Since Brazilian propolis shows an immunomodulatory activity, the goal of this work was to evaluate the action of B. dracunculifolia extracts and some of its isolated compounds on reactive oxygen intermediate (H(2)O(2)) production by macrophages obtained from male BALB/c mice. The results showed that the leaf (Bd-L) (25, 50, and 100 microg mL(-1)), leaf rinse (Bd-LR) (25 microg mL(-1)), and the root (Bd-R) (25 microg mL(-1)) extracts enhanced H2O2 release by macrophages. A phytochemical study of the root and leaves of B. dracunculifolia was carried out. The chromatographic fractionation of Bd-R, using several techniques, afforded the isolation of baccharis oxide (1), friedelanol (2), viscidone (11), 11-hydroxy-10,11-dihydro-euparin (12), and 6hydroxy-tremetona (13), while Bd-LR gave the following isolated compounds: baccharis oxide (1), friedelanol (2), isosakuranetin (3), aromadendrin-4'-methyl ether (4), dihydrocumaric acid (5), baccharin (6), hautriwaic acid lactone (7), hautriwaic acid acetate (8), drupanin (9), and cumaric acid (10). Among the isolated compounds, baccharis oxide (1) and friedelanol (2) increased H2O2 production at a concentration of 100 microM. This is the first time that the presence of compounds 7, 8, 12, and 13 in B. dracunculifolia has been reported. Based on these results it is suggested that the crude extracts and some isolated compounds from B. dracunculifolia display an immunomodulatory action.     

Effect of structural modification of the hydantoin ring on anticonvulsant activity

Selectively substituted hydantoins 1 (15 examples), 4-hydroxy-2-imidazolidinones 2 (13 examples), 2-imidazolones 3 (10 examples), 2-imidazolidinones 4 (four examples), vicinal diamines 5 (two examples), and simple amino acid derivatives 6 (four examples) have been prepared and evaluated in the maximal electroshock seizure (MES), subcutaneous pentylenetetrazole seizure threshold (sc Met), and rotorod (Tox) tests. The medium effective doses (ED50) and the medium toxic dose (TD50) for the most active compounds are reported. In general, the most pronounced activity was observed for hydantoins 1 and protected amino acids 6. Within each series of compounds, enhanced anticonvulsant activity was often noted for compounds containing an aromatic group one carbon removed from a nitrogen atom. Among the most active compounds observed were the amino acid derivative N-acetyl-D,L-alanine benzylamide (6d) and the two 2-imidazolones 4-methyl-1-(phenylmethyl)-1,3-dihydro-2H-imidazol-2-one (3e) and 1-phenyl-1,3-dihydro-2H-imidazol-2-one (3g). Compound 6d proved to be slightly more potent in the MES test than phenacemide.     

真菌Penicillium sp. DT-F27次级代谢产物及其活性研究

摘要：目的 一株海洋来源真菌Penicillium sp. DT-F27次级代谢产物及其抗肿瘤活性的研究。方法 采用硅胶柱色谱、反相高效液相制备色谱等方法,对该真菌发酵产物的乙酸乙酯提取物进行分离纯化;采用波普解析方法对化合物的结构进行鉴定;采用MTT法测定化合物对人前列腺癌PC-3细胞的抗肿瘤作用。结果 从真菌Penicillium sp. DT-F27的发酵物中分离得到15个化合物,分别为麦角甾醇（1）,dehydrocyclopeptine（2）,3-O-methylviridicatin（3）,verrucofortine（4）,cyclopenin（5）,cyclo(dehydroala-L-Leu)（6）,cyclopenol（7）,4-hydroxy-2-methoxy acetanilide（8）,trans-(3RS,4RS)-3,4-dihydro-3,4,8-trihydroxynaphthalen-1(2H)-one（9）,cyclo(D-Pro-D-Val)（10）,brevianamide F（11）,cyclo(L-Pro-L-Val)（12）,anacine（13）,cyclo(L-Orn-L-Tyr-L-Orn-L-Ala)（14）,cyclo(L-Pro-L-Tyr)（15）。结论 从真菌Penicillium sp. DT-F27的次级代谢产物中分离15化合物,其中麦角甾醇（1）和3-O-methylviridicatin（3）具有较强的抗肿瘤活性,抑制率分别为45.6%和34.8%。     

Intestinal permeability of the constituents from the roots of Saposhnikovia divaricata in the human Caco-2 cell monolayer model

The bidirectional intestinal permeability of the active constituents from the roots of Saposhnikovia divaricata, including four coumarins, anomalin (1), 5-methoxy-7-(3,3-dimethylallyloxy)coumarin (2), decursin (3), and decursinol angelate (4), as well as four chromones, cimifugin (5), prim-O-glucosylcimifugin (6), 3'- O-angeloylhamaudol (7), and sec-O-glucosylhamaudol (8), was studied by using the Caco-2 cell monolayer. These compounds were assayed by HPLC, and their transport parameters, including apparent permeability coefficients (P(app)), were then calculated. The bidirectional P(app) values of the compounds were compared with those of the markers, propranolol and atenolol. Compounds 1-5 and 7 were assigned to well-absorbed compounds, while 6 and 8 were assigned to moderately absorbed compounds. The transport of 1-7 increased linearly as a function of time up to 180?min and concentration within the test range of 10-200?μM, thus their passive diffusion mechanism was proposed. The results provided some useful information for predicting the intestinal absorption in vivo of these compounds.     

A New Minor Saponin from the Leaves of Panax ginseng

Seventeen compounds were isolated from the leaves of PANAX GINSENG C. A. Meyer. Among them, a new minor saponin was established as 3beta,6alpha,12beta-trihydroxy-dammar-20(22), 24-diene-6- O-alpha- L-rhamno-pyranosyl-(1-->2)-beta- D-glucopyranoside ( 2). Fourteen compounds were identified as 20( R)-protopanaxadiol ( 1), 20( R)-protopanaxatriol, ginsenoside-Rh (3), 20( R)-ginsenoside-Rh (2), 20( S)-ginsenoside-Rh (2), ginsenoside-Rh (1), -Rg (3), -Rg (2), -Rg (1), -Re, -Rd, -Rc, -Rb (2), -Rb (1); the others are still under investigation.     

Cellular Effects of Some Metabolic Oxidation Products Pertinent to 4-Ethoxyaniline

Abstract: The toxicity of some metabolic products pertinent to 4-ethoxyaniline in isolated hepatocytes were investigated. The compounds investigated were 4-ethoxynitrosobenzene (1), 4-ethoxy-4′-nitrosodiphenylamine (2), 3,6-bis(4-ethoxy-phenylimino)-4-ethoxy-1,4-cyclohexadienylamine (3), 4-(4-ethoxyphenylimino)-2,3-dimethyl-2,5-cyclohexadiene-1-one (4) and 4-(4-ethoxyphenylimino)-2,6-dimethyl-2,5-cyclohexadiene-1-one (5). Of these, 1, 2 and 3 are oxidation products of 4-ethoxyaniline. Compounds 4 and 5 are dimethyl analogues of previously investigated oxidation product 4-(4-ethoxyphenyl-imino(-2,5-cyclohexadiene-1-one (NEPBQI). Among the investigated compounds, 1 and 2 were the most toxic towards isolated hepatocytes. In hepatocytes treated with compounds 1, 2 and 4, loss of cell viability was also accompanied by surface bleb formation. All compounds except 3 reacted with GSH resulting in depletion of cellular GSH. No formation of GSSG was observed, however. Thus, the GSH depletion was apparently due to conjugate formation rather than oxidation. No superoxide dismutase inhibitable reduction of acetylated cytochrome c was observed, thus none of the compounds undergoes measurable redox cycling.     

Pyrenes and pyrendiones from Uvaria lucida

A chemical investigation of the chloroform extract of the roots of Uvaria ludida Benth. (Annonaceae), an important African traditional medicine, led to the isolation of six new compounds; three pyrenes, 2-hydroxy-1,8-dimethoxypyrene (1), 8-methoxy-1,2-methylenedioxypyrene (2), and 7-hydroxy-8-methoxy-1,2-methylenedioxypyrene (3), two pyrenediones, 2-hydroxy-1,8-pyrenedione (4) and 2-methoxy-1,8-pyrenedione (5), and a sesquiterpene, (-)-10-oxo-isodauc-3-en-15-oic acid (6), together with eight known compounds (7-14). The structural elucidation by spectroscopic studies of the compounds isolated is described. While pyrenes did not exhibit strong cytotoxicity against human promyelocytic leukemia HL-60 cells, pyrenediones showed strong cytotoxicity. The IC(50) of 4 was 70 ng mL(-1), which was close to that of etoposide (IC(50) = 60 ng mL(-1)).