双环醇代谢产物的合成

双环醇是我国具有自主知识产权的新一代抗肝炎药，药理作用机制新颖、多环节综合治疗，对慢性乙型肝炎(HBV)临床疗效显著、副作用少，已实现产业化。经药理实验分离出双环醇的代谢产物M2(4-羟基-4′-甲氧基-2-羟甲基-2′-甲氧羰基-5,6,5′,6′-双亚甲二氧基联苯)和M3(4′-羟基-4-甲氧基-2-羟甲基-2′-甲氧羰基-5,6,5′,6′-双亚甲二氧基联苯)，为进一步研究双环醇的作用机制和保证药物的安全性及有效性，本研究全合成其代谢产物。以苄基分别保护芳香溴代物的羧基和酚羟基，通过分子间不对称Ullmann反应，经催化氢化、硼烷还原制得双环醇的两个代谢产物，经红外、高分辨质谱及氢核磁共振谱确定结构。药理试验结果表明，两个代谢产物对CCl₄肝损伤小鼠ALT升高有降低作用，但作用均弱于双环醇。     

联苯双酯代谢产物的分离和鉴定

给大鼠灌胃联苯双酯后,用TLC法分离尿中代谢产物。发现尿中的主要代谢物为葡萄糖醛酸结合物,将此结合物水解后,用TLC法分离到一种极性较葡萄糖醛酸结合物为弱的代谢物,经光谱分析测定其结构为4-羟基-4′-甲氧基-5,6,5′,6′-二次甲二氧基-2,2′-二甲氧羰基联苯(简称4-去甲联苯双酯)。此外还证实,大鼠灌胃后在肝、肾组织中均以葡萄糖醛酸结合物为主,仪有少量非结合形代谢物和原形药物。     

相转移催化合成3-甲氧基-4,5-次甲二氧基苯甲酸甲酯

3-甲氧基-4,5-次甲二氧基苯甲酸甲酯(Ⅰ)为我国独创的治疗肝炎新药联苯双酯(Ⅱ,4,4′-二甲氧基-5,6,5′,6′-二次甲二氧基-2,2′-二甲氧羰基联苯)的主要中间体。可由如下反应制成:     

联苯双酯治疗病毒性肝炎382例临床疗效观察

用五味子制剂治疗迁慢性肝炎已有很多报告,该药共含有七种有效的降低血清谷丙酶成分。联苯双酯(亦称合三,4,4′-二甲氧基-5,6,5′6′-二次甲二氧基-2,2′-二甲氧羰基联苯)为合成五味子丙素时所得的一种中间产物,结构式为:     

硝克柳胺在大鼠和人肝脏的体外代谢研究

研究硝克柳胺在大鼠肝微粒体和胞浆中的代谢动力学，鉴定硝克柳胺在大鼠和人肝微粒体中的主要代谢产物及参与代谢的药物代谢酶。采用高效液相色谱-紫外检测(HPLC-UV)方法测定大鼠肝微粒体和胞浆中硝克柳胺浓度，应用选择性抑制剂鉴定参与硝克柳胺代谢的药物代谢酶类型，采用液相色谱-串联质谱联用(LC-MS/MS)法分离鉴定硝克柳胺在大鼠和人肝微粒体中的主要代谢产物。硝克柳胺在大鼠和人肝微粒体的主要代谢产物(M₁)为硝基还原产物［3-(3′-羧基-4′-羟基苯胺羰基)-6-氨基-7-羟基-8-甲基香豆素］，大鼠体内(血浆、尿液、胆汁及肝组织)主要代谢产物与M₁一致。硝克柳胺的体外代谢是依赖多个药物代谢酶参与的酶促反应，包括微粒体CYP450还原酶、细胞色素b₅还原酶和CYP2C6以及胞浆NAD(P)H脱氢酶和黄嘌呤氧化酶。     

五味子丙素类似物合成的研究—— Ⅱ.4,4′-二甲氧基-5,6,5′,6′-二次甲二氧基-2,2′-二甲氧羰基联苯及其异构体的合成

在合成五味子丙素及共类似物的过程中,合成一些联苯中间体。其中4,4′-二甲氧基-5,6,5′,6′-二次甲二氧基-,2′-二甲氧羰基联苯经药理筛选具有降高血清谷丙转氨酶效果。本文报道上述联苯及其异构体的合成分离和有关光谱数据。     

百赛诺

[通用名称]bicyclol,双环醇 [化学名称]4,4'-二甲氧基-5,6,5',6'-双(亚甲二氧基)-2-羟甲基-2'-甲氧羰基联苯 [理化性质]本品为白色片,不溶于水. [药理作用]本品对刀豆蛋白A(Con A)引起的小鼠肝细胞核DNA损伤有保护作用[1];对四氯化碳、D-氨基半乳糖胺、对乙酰氨基酚引起的小鼠急性肝损伤的转氨酶升高、小鼠免疫性肝炎的氨酶升高有降低作用[2],肝脏组织病理形态学损害有不同程度的减轻作用;体外试验结果显示,本品对肝癌细胞转染人乙肝病毒的2.2.15细胞株具有抑制HBeAg,HBV-DNA,HBsAg分泌的作用[3～5 ]. [适应证]本品可用于治疗慢性肝炎所致的转氨酶升高.     

联苯双酯代谢产物——4-羟基联苯双酯合成的研究

4-羟基联苯双酯(Ⅱ)为联苯双酯(Ⅰ)的代谢产物。本文报道了2-溴-3,4-亚甲二氧基-5-苄氧基苯甲酸甲酯(6)的合成探讨,并将6与2-溴-3,4-亚甲二氧基-5-甲氧基苯甲酸甲酯(8)进行不对称的Ullmann缩合反应,再氢解制得化合物Ⅱ。     

五味子醇甲的代谢转化

采用动物肝微粒体体外代谢法对五味子醇甲的代谢转化进行了研究。从体外代谢产物中鉴定其主要的三个代谢物为:7,8-顺二羟基五味子酸甲;7,7-顺二羟基-2-去甲基五味子醇甲及7,8-顺二羟基-3-去甲基五味子醇甲。在此基础上,建立了生物体液中五味子醇甲及其代谢物的反相HPLC分析方法,并用此法检测了服药后大鼠的胆汁及尿样,比较了体外代谢与体内代谢的异同。     

用~2H-标记物和GC-MS分离鉴定联苯双酯在大鼠尿中的一个代谢产物

联苯双酯(BDD)是人工合成的五味子丙素类似物,其化学结构为4,4′-二甲氧基-5,6,5′,6′-二次甲二氧基-2.2′-二甲氧羰基联苯,已用于迁、慢性肝炎的治疗。临床证明,该药降血清谷丙转氨酶的作用很强。为阐明其在体内的转化过程,并进一步研究其构效关系,我所曾用放射性同位素结合薄层层析对联苯双酯的代谢途径进行过研究,并分     

Identification of the metabolites of episesamin in rat bile and human liver microsomes

Episesamin is an isomer of sesamin, resulting from the refining process of non-roasted sesame seed oil. Episesamin has two methylendioxyphenyl groups on exo and endo faces of the bicyclic skeleton. The side methylendioxyphenyl group was metabolized by cytochrome-P450. Seven metabolites of episesamin were found in rat bile after treatment with glucuronidase/arylsulfatase and were identified using NMR and MS. The seven metabolites were (7α,7'β,8α,8'α)-3,4-dihydroxy-3',4'-methylenedioxy-7,9':7',9-diepoxylignane (EC-1-1), (7α,7'β,8α,8'α)-3,4-methylenedioxy-3',4'-dihydroxy-7,9':7',9-diepoxylignane (EC-1-2) and (7α,7'β,8α,8'α)-3,4:3',4'-bis(dihydroxy)-7,9':7',9-diepoxylignane (EC-2), (7α,7'β,8α,8'α)-3-methoxy-4-hydroxy-3',4'-methylenedioxy-7,9':7',9-diepoxylignane (EC-1m-1), (7α,7'β,8α,8'α)-3,4-methylenedioxy-3'-methoxy-4'-hydroxy-7,9':7',9-diepoxylignane (EC-1m-2), (7α,7'β,8α,8'α)-3-methoxy-4-hydroxy-3',4'-dihydroxy-7,9':7',9-diepoxylignane (EC-2m-1) and (7α,7'β,8α,8'α)-3,4-dihydroxy-3'-methoxy-4'-hydroxy-7,9':7',9-diepoxylignane (EC-2m-2). EC-1-1, EC-1-2 and EC-2 were also identified as metabolites of episesamin in human liver microsomes. These results suggested that similar metabolic pathways of episesamin could be proposed in rats and humans.     

In vitro metabolism of genistein and tangeretin by human and murine cytochrome P450s

Recombinant cytochrome P450 (CYP) 1A2, 3A4, 2C9 or 2D6 enzymes obtained from Escherichia coli and human liver microsomes samples were used to investigate the ability of human CYP enzymes to metabolize the two dietary flavonoids, genistein and tangeretin. Analysis of the metabolic profile from incubations with genistein and human liver microsomes revealed the production of five different metabolites, of which three were obtained in sufficient amounts to allow a more detailed elucidation of the structure. One of these metabolites was identified as orobol, the 3'-hydroxylated metabolite of genistein. The remaining two metabolites were also hydroxylated metabolites as evidenced by LC/MS. Orobol was the only metabolite formed after incubation with CYP1A2. The two major product peaks after incubation of tangeretin with human microsomes were identical with 4'-hydroxy-5,6,7,8-tetramethoxyflavone and 5,6-dihydroxy-4',7,8-trimethoxyflavone, previously identified in rat urine in our laboratory. By comparison with UV spectra and LC/MS fragmentation patterns of previously obtained standards, the remaining metabolites eluting after 14, 17 and 20 min. were found to be demethylated at the 4',7-, 4',6-positions or hydroxylated at the 3'- and demethylated at the 4'-positions, respectively. Metabolism of tangeretin by recombinant CYP1A2, 3A4, 2D6 and 2C9 resulted in metabolic profiles that qualitatively were identical to those observed in the human microsomes. Inclusion of the CYP1A2 inhibitor fluvoxamine in the incubation mixture with human liver microsomes resulted in potent inhibition of tangeretin and genistein metabolism. Other isozymes-selective CYP inhibitors had only minor effects on tangeretin or genistein metabolism. Overall the presented observations suggest major involvement of CYP1A2 in the hepatic metabolism of these two flavonoids.     

New prenylated metabolites of Deguelia longeracemosa and evaluation of their antimicrobial potential

The analysis of root extracts from Deguelia longeracemosa (Benth.) A.M.G. Azevedo yielded fifteen prenylated metabolites. Nine of them are novel, and their molecular structures were determined through spectral analyses (UV, IR, MS and NMR) as being five derivatives of 4-hydroxy-3-phenylcoumarin: 4-hydroxy-3-(4'-hydroxyphenyl)-5-methoxy-6-(8',9'-epoxy-9'-methylbutyl)-2',2'-dimethylpyrano-(5',6':8,7)-coumarin; 4-hydroxy-3-(3',4'-methylenedioxyphenyl)-5-methoxy-6-(3,3-dimethylallyl)-2',2'-dimethypyrano-(5',6':8,7)-coumarin; 4-hydroxy-3-(3'-hydroxy-4'-methoxyphenyl)-5-methoxy-6-(3,3-dimethylallyl)-2',2'-dimethylpyrano-(5',6':8,7)-coumarin; 4-hydroxy-3-(3'-hydroxy-4'-methoxyphenyl)-5-methoxy-2',2'-dimethylpyrano-(5',6':6,7)-coumarin and 4-hydroxy-3-[4'-O-(3,3-dimethylallyl)phenyl]-5-methoxy-2',2'-dimethylpyrano-(5',6':6,7)-coumarin, three derivatives of 1,2-diphenyl-1,2-ethanodione (alpha-oxodeoxybenzoin derivatives): 1-[6-hydroxy-2-methoxy-3-(3,3-dimethylallyl)-2',2'-dimethylpyrano-(5',6':5,4)- ]-2-(4'-hydroxyphenyl)-1,2-ethanedione; 1-[6-hydroxy-2-methoxy-2',2'-dimethylpyrano-(5',6':3,4)]-2-(4'-methoxyphenyl)-1,2-ethanedione; 1-[6-hydroxy-2-methoxy-2',2'-dimethylpyrano-(5',6':3,4)]-2-(3',4'-methylenedioxyphenyl)-1,2-ethanedione and one derivative of deoxybenzoin: 2,4'-dimethoxy-6-hydroxy-2',2'-dimethylpyrano-(5',6':3,4)-deoxybenzoin. The antimicrobial activity of roots extracts and some isolated compounds was screened through bioautography against bacteria and fungi.     

Dihydrochalcones from Piper longicaudatum

Bioactivity-guided fractionation of an ethanolic extract of the leaves and twigs of Piper longicaudatum Trelease & Yunker (Piperaceae) resulted in the isolation of one new (1) and three known (2-4) dihydrochalcones. The known compounds are: 2',6'-dihydroxy-4'-methoxydihydrochalcone (2), 2',6',4-trihydroxy-4'-methoxydihydrochalcone (asebogenin) (3), and 2'-hydroxy-4'-methoxy-2'-[1-hydroxy-1-methylethyl]-2",3"-dihy- drofurano[4",5":5',6"]-3"-[2-hydroxy-5-methoxycarbonylphe- nyl]dihydrochalcone (piperaduncin B) (4). The new compound is 2'-hydroxy-4'-methoxy-2"-[2-hydroxy-5-methoxycarbonyl- phenyl]-furano[4",5":5',6']-dihydrochalcone (longicaudatin) (1). Compounds 1-4 were tested for antibacterial activity against S. aureus and methicillin-resistant S. aureus (MRSA); only compound 3 showed inhibitory activity (IC50 of 10 and 4.5 micrograms/ml, respectively).     

Metabolism of pigment yellow 74 by rat and human microsomal proteins.

Pigment Yellow 74 (PY74) is a monoazo pigment that is used in yellow tattoo inks. The metabolism of PY74 was investigated using rat liver and human liver microsomes and expressed human cytochromes P450 (P450s). Two phase I metabolites were isolated and characterized by mass spectrometry and NMR techniques. One metabolite (PY74-M1) was a ring hydroxylation product of PY74, 2-((2-methoxy-4-nitrophenyl)azo)-N-(2-methoxy-4-hydroxyphenyl)-3-oxobutanamide. The second metabolite (PY74-M2) was identified as 2-((2-hydroxy-4-nitrophenyl)azo)-N-(2-methoxy-4-hydroxyphenyl)-3-oxobutanamide, which is the O-demethylation product of PY74-M1. These metabolites were formed by in vitro incubations of PY74 with 3-methylcholanthrene-induced rat liver microsomes and to a much lesser extent by liver microsomes from untreated or phenobarbital-induced rats. The role for CYP1A in the metabolism of PY74 was confirmed using expressed human P450s. The catalytic ability of the P450s for metabolism of PY74 was CYP 1A2 > CYP 1A1 > CYP 3A4 approximately CYP 1B1 (no activity with CYP 2B6, 2C9, 2D6 or 2E1). The metabolism of PY74-M1 to PY74-M2 was catalyzed only by CYP 1A2 and CYP 1A1 (no activity from CYP 1B1, 2B6, 2C9, 2D6, 2E1, or 3A4). These results demonstrate that the tattoo pigment PY74 is metabolized in vitro by P450 to metabolites that should be available for phase II metabolism and excretion.     

In vitro metabolism of 2,2',3,4',5,5',6-heptachlorobiphenyl (CB187) by liver microsomes from rats, hamsters and guinea pigs

The metabolism of 2,2',3,4',5,5',6-heptachlorobiphenyl (heptaCB) (CB187) was studied using liver microsomes of rats, hamsters and guinea pigs, and the effect of cytochrome P450 (CYP) inducers, phenobarbital (PB) and 3-methylcholanthrene (MC), was also investigated. In untreated animals, guinea pig liver microsomes formed three metabolites which were deduced to be 4'-hydroxy-2,2',3,5,5',6-hexachlorobiphenyl (M-1), 4'-hydroxy-2,2',3,3',5,5',6-heptaCB (M-2) and 4-OH-CB187 (M-3) from the comparison of GC/MS data with some synthetic authentic samples. The formation rate of M-1, M-2 and M-3 was 18.1, 36.6, 14.7 pmol h-1 mg protein-1, respectively. Liver microsomes of untreated rats and hamsters did not form CB187 metabolites. In guinea pigs, PB-treatment increased M-1 and M-2 significantly to 1.9- and 3.4-fold of untreated animals but did not affect the formation of M-3. In rats, PB-treatment resulted in the appearance of M-2 and M-3 with formation rates of 87.1 and 13.7 pmol h-1 mg protein-1, respectively, but M-1 was not observed. In hamsters, PB-treatment formed only M-2 at a rate of 29.4 pmol h-1 mg protein-1. On the other hand, MC-treatment of guinea pigs decreased the formation of M-1 and M-2 to less than 50% of untreated animals. MC-microsomes of rats and hamsters produced no metabolites. Preincubation of antiserum (300 microl) against guinea pig CYP2B18 with liver microsomes of PB-treated guinea pigs produced 80% inhibition of M-1 and the complete inhibition of M-2 and M-3. These results suggest that PB-inducible CYP forms, especially guinea pig CYP2B18, rat CYP2B1 and hamster CYP2B, are important in CB187 metabolism and that CB187 metabolism in guinea pigs may proceed via the formation of 3,4- or 3',4'-oxide and subsequent NIH-shift or dechlorination.     

Two dihydropyrrolizine alkaloid metabolites isolated from mouse hepatic microsomes in vitro

The in vitro mouse hepatic microsomal metabolism of the macrocyclic pyrrolizidine alkaloid senecionine was studied for additional metabolites. Using previously developed HPLC systems plus a preparative system, two additional dihydropyrrolizine metabolites have been identified from the microsomal enzyme system of mice. The metabolites 1-hydroxymethyl-7-methoxy-6,7-dihydro-5H-pyrrolizine (methoxydehydroretronecine) and 1-formyl-7-hydroxy-6,7-dihydro-5H-pyrrolizine (hydroxydanaidal) have not been heretofore isolated from mouse microsomal enzyme systems. The metabolite dehydroretronecine which had previously been isolated from rat hepatic microsomes, was not detected while senecic acid, 19-hydroxysenecionine, and senecionine N-oxide were again present.     

In vitro biotransformation of 3,4-dihydro-6-hydroxy-2,2-dimethyl-7-methoxy-1(2H)-benzopyran (CR-6), a potent lipid peroxidation inhibitor and nitric oxide scavenger, in rat liver microsomes

The in vitro metabolism of 3,4-dihydro-6-hydroxy-2,2-dimethyl-7-methoxy-1(2H)-benzopyran (CR-6), a potent lipid peroxidation inhibitor and scavenger of nitric oxide and peroxynitrite species that is currently in phase II trials for antitumoral therapy, has been investigated in rat liver microsomes in the presence of NADP(H). Five major metabolites were identified by comparison with authentic standards, namely, the quinone 2-(3'-hydroxy-3'-methylbutyl-5-methoxy-1,4-benzoquinone (2a) and its ring-closed spiro form oxaspiro[4.5]-2,2-dimethyl-8-methoxy-dec-8-ene-7,10-dione (2b), the hydroquinone 2-(3'-hydroxy-3'-methylbutyl)-5-methoxyhydroquinone (3), the hydroxylated metabolite 3,4-dihydro-4,6-dihydroxy-2,2-dimethyl-7-methoxy-1(2H)-benzopyran (4), and the catechol 3,4-dihydro-6,7-dihydroxy-2,2-dimethyl-1(2H)-benzopyran (5). When the incubations were carried out in the presence of GSH, the HPLC peaks corresponding to the quinone metabolites 2a/b were absent and two novel products were formed showing MS fragmentation patterns consistent with the structure of GSH conjugates of quinone 2a. The time dependence on the formation of metabolites 2a,b and 3 was measured in incubations induced with phenobarbital (PB), dexamethasone, and beta-naphthoflavone (betaNF). For the dexamethasone-induced microsomes, the amount of hydroquinone 3 decreased from minute 10 to minute 30 while that of 2a,b increased in a complementary manner. Similar effects were observed for the incubations carried out using PB- and betaNF-induced microsomes. On the other hand, CR-6 inhibited 7-ethoxyresorufin O-dealkylation activity (IC(50) = 25 microM) in incubations with betaNF-induced microsomes. Likewise, addition of pentoxyresorufin to the incubations of CR-6 with PB-induced microsomes showed a time-dependent inhibition (IC(50)= 75 microM) of the dealkylation activity. These results are in agreement with the putative generation of reactive metabolites from CR-6 that could deactivate P450 1A and P450 2B, respectively. When these incubations were carried out in the presence of 10 mM GSH, the inhibition of P450 2B could be partially prevented. Finally, preincubation of CR-6 with liver microsomes from PB-induced rats resulted in a strong increase in microsomal glutathione S-transferase (mGST) activity (up to a maximum of approximately 5-fold). When the preincubation was carried out in the presence of 10 mM GSH, the activation of mGST was blocked. Overall, these results suggest that CR-6 undergoes in vitro biotransformation indicative of the involvement of thiol-reactive metabolites.     

In vitro metabolism of the antianxiety drug buspirone as a predictor of its metabolism in vivo

1. Metabolism of the antianxiety drug buspirone was studied by in vitro incubations with rat liver microsomes and hepatocytes. Metabolites were isolated and purified by h.p.l.c. The purified metabolites were identified by co-elution on h.p.l.c. with authentic standards and by g.l.c.-electron impact mass spectrometry of their trimethylsilyl (TMS) derivatives. 2. Five metabolites of buspirone were identified in the microsomal incubates and seven in the hepatocyte incubates. The major metabolites arose from aromatic hydroxylation at C-5, N-dealkylation of the butyl chain, and hydroxylation at C-6' and C-3' on the azaspirodecanedione moiety. 3. Metabolism of buspirone by rat liver microsomes was NADPH-dependent and was completely inhibited by cytochrome P-450 inhibitors SKF-525A and metyrapone. 4. Metabolites of buspirone formed in vitro were good predictors of the primary metabolites formed in vivo. 5. Hepatocytes and phenobarbital-induced rat liver microsomes were better predictors of in vivo metabolism of buspirone than non-induced rat liver microsomes. These in vitro systems should provide excellent models for studying the metabolism of other azaspirodecanedione-containing drugs.