Isolation of dolastatin 10 from the marine cyanobacterium Symploca species VP642 and total stereochemistry and biological evaluation of its analogue symplostatin 1

The potent antitumor agent dolastatin 10 (1) was originally isolated from the sea hare Dolabella auricularia, and we now report its isolation from the marine cyanobacterium Symploca sp. VP642 from Palau. The chemically related analogue symplostatin 1 (2) has been reisolated from Guamanian and Hawaiian varieties of S. hydnoides and its total stereochemistry completed by determining the N,N-dimethylisoleucine unit to be L. Symplostatin 1 (2), like dolastatin 10 (1), is a potent microtubule inhibitor. The antitumor activity of 2 was assessed in vivo against several murine tumors. Symplostatin 1 (2) was effective against a drug-insensitive mammary tumor and a drug-insensitive colon tumor; however, it was only slightly effective against two MDR tumors.     

Antitumor agents. 190. Absolute stereochemistry of the cytotoxic germacranolides, tomenphantins A and B, from Elephantopus tomentosus

The structures and absolute stereochemistries of tomenphantins A (1) and B (2), cytotoxic germacranolides isolated from Elephantopus tomentosus, are reported herein. 1H and 13C NMR spectroscopic data, chemical transformation, and single-crystal X-ray analysis were used in these determinations.     

丹参中的一个新化合物及其抗HIV活性

目的研究甘西鼠尾草Salvia przewalskii根的化学成分，检测抗HIV活性。方法用溶媒法、色谱法提取分离单体，光谱法分析鉴定结构。用艾滋病毒1型整合酶(HIV-1 IN)和艾滋病毒1型逆转录酶(HIV-1 RT)测定抗HIV活性。结果分离得到4个化合物，分别鉴别为8-{[7‘-(3’，‘4-二羟基-苯基)-9‘-氧代-7’-丙烯基]氧)-3-(1“-O-β-D-葡萄糖基)-4-羟基-[R-(E)]-苯丙酸(I)，迷迭香酸(Ⅱ)，紫草酸(Ⅲ)和紫草酸B(IV)。结论I为新化合物，Ⅱ～IV为已知化合物。均具有抗HIV活性。     

Isolation and identification of a cannabinoid-like compound from Amorpha species

The petroleum ether extracts from several plants have been screened on the plates with the chromogenic reagent Fast Blue B. salt. Extracts from different plant parts of Amorpha fruticosa, A. nanna, and A. canescens gave several spots with color and Rf values similar to those of Cannabis sativa extract. The major compound, amorphastilbol (1), was isolated by column chromatography, purified, and characterized by mp, elemental analysis, uv, ir, mass, prm, and cmr spectroscopy. Further information was obtained by analyzing the oxidation products of the compound. The evidence indicates that amorphastilbol is a phenolic stilbene terpenoid with a molecular formula of C24 H28 O2 and structural formula: (E,E)-2-(3',7'-dimethyl-2'-6'-octadienyl)-5-(2-phenylethenyl)-1,3-benzenediol. This is the first report of the isolation of a naturally-occurring stilbene derivative possessing a terpenoid moiety.     

Antiulcer and gastric antisecretory effects of dichloromethane fraction and piplartine obtained from fruits of Piper tuberculatum Jacq. in rats

Ethnopharmacological relevance

Piper tuberculatum Jacq. (Piperaceae) is medicinally used as an analgesic and as a treatment for gastric complaints. Thus, the current study aimed to investigate the gastroprotective and antisecretory properties of the dichloromethane fraction of the fruit of Piper tuberculatum (DFPT) and piplartine, a compound isolated from the DFPT, in rats.

Materials and methods

Gastric ulcers were induced in fasted rats by oral administration of absolute ethanol and then mucus content and glutathione (GSH) levels were measured. Mechanisms underlying the antisecretory action were studied through gastric H⁺,K⁺-ATPase activity of highly purified rabbit gastric microsomes and pylorus ligature method in rats.

Results

In the acute toxicity test the values of estimated LD₅₀ for oral and intraperitoneal administration of DFPT were 1.6266 and 0.2684 g/kg, respectively. The DFPT (ED₅₀=29 mg/kg, p.o.) and piplartine (4.5 mg/kg, p.o.) promoted gastroprotection against acute lesions induced by ethanol, effect that could be related with the maintenance of GSH levels in the gastric mucosa. However, only DFPT stimulated gastric mucus secretion. In vitro, the DFPT and piplartine inhibited the H⁺,K⁺-ATPase activity and, in vivo DFPT and piplartine also reduced basal gastric acid secretion, as well as that stimulated by pentagastrin.

Conclusions

These results demonstrate that DFPT and piplatine cause marked gastroprotective effects accompanied by the increase and maintenance of gastric mucus and GSH levels, as well as a reduction in gastric acid secretion through the gastrinergic pathway.     

补骨脂中1个二氢异黄酮类新化合物及其细胞毒活性

目的 对中药补骨脂Psoralea corylifolia中的化学成分进行研究, 以期发现新的活性化合物。方法 运用硅胶、凝胶、MCI-GEL树脂及RP-HPLC等多种色谱技术对补骨脂的化学成分进行分离纯化, 并根据理化性质和波谱数据鉴定化合物的结构。结果 从补骨脂95%乙醇提取物分离鉴定了1个二氢异黄酮类化合物3R-6-羟基-8-羟甲基-4',7-二甲氧基-二氢异黄酮(1)。结论 化合物1为新化合物, 命名为补骨脂异黄酮A(corylisoflavone A);表现出一定的细胞毒活性, 其中对急性早幼粒细胞白血病细胞(NB4)、人神经母细胞瘤细胞(SH-SY5Y)、人前列腺癌细胞(PC3)、人肺腺癌细胞(A549)和人乳腺癌细胞(MCF7)的IC₅₀值分别为12.6、5.3、15.5、4.8和11.2 μmol/L。     

金铃子散不同配比方对大鼠肝药酶CYP1A2活性的影响

目的:通过比较金铃子散不同配比方对大鼠肝药酶CYP1A2活性的影响,探讨其配伍规律。方法:按L9(34)正交表,设计9个不同配比组方。体外实验采用大鼠肝药酶孵育体系,测定不同配比方对CYP1A2的半数抑制浓度(IC50);体内实验采用大鼠口服金铃子散不同配比方5 d后注射探针药物非那西丁,通过微透析探针采集肝脏部位样品,HPLC测定非那西丁及其代谢物对乙酰氨基酚浓度,winNonlin软件统计拟合药代参数。结果:川楝子、延胡索单味提取物和配比方1～9对肝药酶CYP1A2的IC50分别为(0.025 2±0.005 2),(0.012 1±0.007 9),(0.091 9±0.015 0),(0.071 9±0.005 3),(0.028 2±0.004 5),(0.075 4±0.015 5),(0.062 8±0.003 3),(0.091 9±0.015 0),(0.197 6±0.027 3),(0.159 1±0.008 1),(0.131 1±0.008 5)g.L-1,无显著抑制酶CYP1A2活性。体内实验,药代参数显示金铃子散不同配比方均能诱导CYP1A2活性,影响酶活性强弱顺序是:诱导剂组(方3)>方4(方5)>方9(方6,方1,方2,方8,川楝子,延胡索组)>方7>正常对照组。结论:金铃子散不同配比方对肝药酶CYP1A2活性有诱导作用,且诱导作用的强弱与配比有一定的相关性。     

化浊解毒复方对中晚期胃癌血清血管内皮细胞生长因子及环氧化酶-2的影响

目的 观察化浊解毒复方对胃癌血清血管内皮细胞生长因子(VEGF)和环氧化酶-2(COX -2)的影响.方法 将中晚期胃癌66例随机分为2组,均予FOLFOX4方案化疗.治疗组同时予化浊解毒复方,日1剂,水煎服.以14 d为1个周期,均连续治疗4个周期.2组分别于治疗前、治疗第2、4个周期后观察疗效、生存质量,并检测...     

On the anti-inflammatory and anti-phospholipase A(2) activity of extracts from lanostane-rich species

We have studied extracts from three species rich in lanostane triterpenes for their activity against different in vivo models of inflammation induced by TPA, EPP and PLA(2). The inhibitory effect against PLA(2) in vitro was also studied. When the Poria cocos extract was tested against PLA(2)-induced mouse paw edema, it was active by the oral and parenteral routes. Its effect was greater in both magnitude and duration than that of Pistacia terebinthus and Ganoderma lucidum extracts. P. terebinthus was effective against chronic and acute inflammation, and according to a preliminary chromatographic analysis, its seems to be a good source of lanostane anti-inflammatory agents. G. lucidum was the least effective of the three species studied and, unlike the other two, failed to inhibit the activity of PLA(2) in vitro.     

胃黏膜损伤大鼠胃组织P2X_1、P2X_3和P2X_5的表达及电针对其影响

目的观察电针足三里穴对胃黏膜应激损伤大鼠胃组织中P2X1、P2X3、P2X5mRNA表达的影响,探讨电针足三里穴防治胃黏膜损伤的分子机制。方法将SD大鼠随机分为正常对照组(正常组)、胃黏膜应激损伤模型组(模型组)和电针足三里穴组(电针组)。采用冷-束缚法制作胃黏膜应激损伤大鼠模型。造模后次日起电针组每天电针足三里穴20分钟,连续三天。RT-PCR法检测胃组织P2X1、P2X3、P2X5mRNA表达。结果各组大鼠均表达P2X1、P2X3、P2X5mRNA;模型组与正常组比较,P2X1mRNA表达显著降低(P0.01);电针组与模型组比较,P2X1mRNA表达显著升高(P0.05);P2X3、P2X5mRNA组间未见显著差异;正常组大鼠胃组织P2X3和P2X5呈显著正相关(P0.05)。结论胃黏膜应激损伤大鼠的胃组织P2X1mRNA表达下降,电针足三里穴可以上调胃黏膜应激损伤大鼠胃组织P2X1mRNA的表达。     

狗肝菜多糖P1A的结构解析

采用完全酸水解、部分酸水解、高碘酸氧化-Smith降解、甲基化等方法,结合红外光谱和核磁共振分析对P1A进行结构解析.结果表明,P1A的主链由鼠李糖(Rha)、甘露糖(Man)、葡萄糖(Glc)和半乳糖(Gal)组成,支链包含阿拉伯糖(Ara)和木糖(Xyl).P1A中含有1→,1→6键型和非还原末端,也含有1→2和1→4键型,这2种键型比例约为1:2.可氧化键型占45％,不可氧化键型占55％.P1A中Man,Glc,Gal主要以1→3键型相连,Rha,Ara,Xyl主要以1→2和1→4键型相连.P1A中含有以下片段β-Glc(1,6)-,β-Gal(1,3)-,β-Man(1,4)-β-Rha,-Glc(1,4)-,Glc(1)-,-Gal(1,4)-,Man(1)-.     

相思子蛋白P2对B16黑色素瘤的抑制作用及机制研究

目的:研究相思子蛋白P2对小鼠黑色素瘤B16生长的抑制作用及作用机制。方法:MTT法观察相思子蛋白P2对B16细胞增殖的抑制作用,流式细胞仪检测对B16细胞周期和细胞凋亡的影响,JC-1染色法检测相思子蛋白P2对B16细胞线粒体膜电位的影响;相思子蛋白P2 50、75、100μg/kg连续灌胃给药10天,观察对小鼠黑色素瘤B16移植性肿瘤生长的抑制作用。结果:相思子蛋白P2对B16细胞具有显著的生长抑制作用,IC50值为4.6×10-3μg/ml,浓度为1μg/ml时抑制率可达95.45%;流式细胞仪分析显示,相思子蛋白P2主要将细胞周期滞留在S期,阻断向G2/M期发展,从而抑制肿瘤细胞增殖。相思子蛋白P2可明显诱导B16细胞凋亡。荧光显微镜检测结果显示相思子蛋白P2可显著降低B16细胞线粒体膜电位。相思子蛋白P2口服给药对小鼠B16移植性肿瘤生长有抑制作用,100μg/kg抑瘤率达53.75%,有明显剂量-效应依赖关系,且对胸腺和脾脏质量指数的影响较环磷酰胺小。结论:相思子蛋白P2于体内外均可显著抑制B16细胞的生长,此作用与阻止肿瘤细胞增殖周期,降低线粒体膜电位,诱导细胞凋亡有关。     

The enzymatic decomposition of salicin and its derivatives obtained from Salicaceae species.

The enzymatic catalysis of the decomposition of Salicaceae phenolic glucosides was tested using almond beta-glucosidase and rabbit and porcine liver esterases. The beta-glucosidase catalyzed the complete hydrolysis of salicin and salicortin, yielding saligenin and glucose. Salicortin also produced (+)-6-hydroxycyclohexen-2-one (6-HCH). The acylglucosides were not decomposed by the beta-glucosidase. Both esterases catalyzed the decomposition of tremulacin, salicortin, and 2'-O-acetylsalicortin, releasing tremuloidin, salicin, and 2'-O-acetylsalicin as the main products, accompanied by 6-HCH and catechol. Tremuloidin and 2'-O-acetylsalicin were quite stable under the esterase hydrolysis, and salicin was not decomposed at all.     

傣药翅荚决明细枝中一个新的2-芳基苯并呋喃类化合物及细胞毒活性

目的对傣药翅荚决明Cassia alata细枝的化学成分进行研究,以期发现新的活性化合物。方法运用硅胶、凝胶、MCI-GEL树脂及RP-HPLC等多种色谱技术进行分离纯化,根据理化性质和波谱数据鉴定化合物的结构,并用MTT法测定了化合物对NB4、A-549、SHSY5Y、PC-3和MCF-7 5种人源肿瘤细胞株的细胞毒活性。结果从翅荚决明细枝中分离鉴定了4个2-芳基苯并呋喃类化合物,分别鉴定为7-甲氧基-2-(4-甲氧基苯基)-3,5-二甲基苯并呋喃(1)、桑辛素N(2)、2-(2′-甲氧基-4′-羟基苯基)-3-甲基-6-羟基苯并呋喃(3)、桑辛素P(4)。结论化合物1为新化合物,命名为翅荚决明素A,该化合物对所测试的人源肿瘤细胞具有一定的细胞毒活性。     

加味升降散对病毒感染后小鼠呼吸道上皮VR1表达及肺内P物质含量的影响

目的探讨加味升降散对病毒感染后小鼠呼吸道上皮辣椒素受体(VR1)表达及P物质含量的影响。方法将40只SPF级小鼠随机分为治疗组与对照组,均采用甲型流感病毒FMl株造模,造模成功后治疗组予加味升降散灌胃,对照组予蒸馏水灌胃,连续2周。末次灌胃后24 h处死小鼠,电镜下观察呼吸道上皮病理变化,PCR法检测呼吸道上皮VR1mRNA表达量,ELISA法检测肺组织匀浆上清液中P物质含量。结果治疗组小鼠呼吸道上皮VR1 mRNA的表达量及肺组织匀浆P物质含量均明显低于对照组(P均0.05),并且呼吸道上皮纤毛有修复现象。结论加味升降散对病毒感染后小鼠呼吸道上皮具有修复作用。     

细胞色素P450 1A2和2C19基因多态性与抗结核药物性肝损伤的关系

 目的 探讨细胞色素P450 1A2和2C19基因多态性与抗结核药物性肝损伤的相关性。方法 采用1∶1病例对照研究,以抗结核药治疗致肝损伤的结核病患者为病例组,无肝损伤者为对照组,共179对。采用聚合酶链反应限制性片段长度多态方法检测CYP1A2基因734C/A和CYP2C19基因681G/A位点基因型。采用SPSS17.0条件Logistic回归进行数据分析。结果 CYP1A2 基因734C/A位点C、A等位基因频率在病例组和对照组频率分别为37.2%、62.8%、和47.2%、52.8%;CYP2C19基因的681G/A位点G、A等位基因频率在病例组和对照组频率分别为59.8%、40.2%和62.8%、37.2%,CYP1A2基因734C/A位点多态性与抗结核药物性肝损伤的发生有关(P＜0.05),CA、AA基因型的比值比(odds ratio,OR)分别为1.951和1.916, 而尚不能认为CYP2C19基因681G/A位点多态性与抗结核药物性肝损伤发生有关。结论 CYP1A2基因734C/A位点多态性与抗结核药物性肝损伤的发生有关,并且携带CA、AA突变基因型的个体是危险人群。     

Luteolin, a compound with adenosine A(1) receptor-binding activity, and chromone and dihydronaphthalenone constituents from Senna siamea

Activity-guided fractionation led to the isolation of luteolin (1) from the leaves of Senna siamea (syn. Cassia siamea). This compound was found to be an antagonist at the adenosine A(1) receptor with a K(i) value in the low micromolar range. Four additional nonactive compounds (2-5) were also isolated, and their structures were elucidated. One compound was identified as cassia chromone (5-acetonyl-7-hydroxy-2-methylchromone) (2). Three other compounds are new, and they were identified as 5-acetonyl-7-hydroxy-2-hydroxymethyl-chromone (3), 4-(trans)-acetyl-3,6,8-trihydroxy-3-methyldihydronaphthalenone (4), and 4-(cis)-acetyl-3,6,8-trihydroxy-3-methyldihydronaphthalenone (5).     

羟基芫花素对UGTs及UGT1A1活性抑制作用种属差异的体外研究

考察羟基芫花素对尿苷二磷酸葡萄糖醛酸转移酶(UGTs)及UGT1A1活性的影响,为预测其与其他药物的代谢性相互作用提供理论借鉴。该实验采用体外肝微粒体孵育模型,以4-硝基酚(4-NP)为底物检测UGTs活性;β-雌二醇为底物检测UGT1A1活性,利用UV和HPLC测定底物或代谢物含量。结果表明,在大鼠、小鼠和人肝微粒体(HLM)孵育体系,羟基芫花素能显著抑制UGTs活性;对UGT1A1,在小鼠肝微粒体(MLM)孵育体系中,羟基芫花素几乎无抑制作用(IC50=190μmol·L-1);在大鼠肝微粒体(RLM)和重组酶(r UGT1A1)孵育体系中,羟基芫花素表现为中等强度的抑制作用(IC50=10.93,20.07μmol·L-1),抑制类型分别为竞争性抑制和线性混合型抑制;在HLM孵育体系中,羟基芫花素表现为弱抑制作用(IC50=76.31μmol·L-1),抑制类型为竞争性抑制;其抑制强弱顺序为RLMr UGT1A1HLMMLM。综上,羟基芫花素对不同肝微粒体孵育体系中UGTs及UGT1A1活性均可产生抑制作用且存在种属差异性,提示羟基芫花素可能存在基于UGT1A1酶的药物相互作用。该研究可为合理开发利用羟基芫花素提供实验依据,并为研究药物在临床上的联合用药提供理论借鉴。     

复方苦参注射液预防胃癌术后患者化疗不良反应及对免疫功能的影响

目的探讨复方苦参注射液对胃癌术后患者化疗不良反应的防治作用及对免疫功能的影响。方法将99例胃癌术后化疗患者按就诊先后顺序随机分为2组。治疗组49例,在常规化疗基础上加用复方苦参注射液治疗;对照组50例,单纯予常规化疗治疗。观察比较2组治疗后出现不良反应情况,包括白细胞计数(WBC)降低、血小板计数(PLT)降低、血红蛋白(Hgb)降低、谷氨酸氨基转移酶(ALT)升高及出现恶心呕吐、腹泻、口腔炎、神经毒性患者的例数;比较2组治疗前及治疗1周后免疫功能变化,包括自然杀伤(NK)细胞及T淋巴细胞亚群(CD3+、CD4+、CD8+及CD4+/CD8+)水平变化。结果治疗组治疗后WBC降低、PLT降低、ALT升高及恶心呕吐发生情况均低于对照组(P0.05),Hgb降低、腹泻、口腔炎及神经毒性发生情况与对照组比较差异无统计学意义(P0.05);2组治疗前各免疫功能指标比较差异无统计学意义(P0.05),治疗组治疗1周后各免疫指标与治疗前比较水平无明显变化(P0.05),对照组治疗1周后CD3+、CD4+、CD4+/CD8+及NK细胞水平与治疗前比较均有明显降低(P0.05)。结论复方苦参注射液可以明显减少胃癌术后化疗患者不良反应的发生,并提高患者免疫功能,改善患者临床症状,提高患者生活质量,增强患者化疗耐受能力。     

以非那西丁为探针采用高效液相色谱法快速测定细胞色素P450 1A2的酶活性

目的 建立一种快速、高效的以非那西丁作为探针药物评价细胞色素P450 1A2(CYP1A2)酶活性的高效液相色谱-紫外检测方法.方法 色谱柱为Agilent Zorbax SB-C18柱(50 mm×4.6 mm I.D.,5μm);流动相为乙腈-水(含0.01%七氟丁酸),梯度洗脱,流速2.0 ml·min-1;检测波长为244 nm.非那西丁与人CYP1A2酶在37℃温孵适当时间后,加入50 μl乙腈终止反应,10 000 g离心后取上清液进样分析测定.结果 对乙酰氨基酚的保留时间为2.85 min,线性范围为1.00～200 μmol·L-1(r=0.999 9),最低定量限(LLOQ)为1.00 μmol·L-1,回收率为99.8%～102.3%;非那西丁的睬留时间为3.68 min,线性范围为1.00～200 μmol·L-1(r=0.999 9),最低定量限(LLOQ)为1.00 μmol·L-1,回收率为98.3%～101.1%.两者的日内、日间相对标准偏差均小于15%,温孵体系中的其他内源性物质不干扰测定.结论 该方法快速、稳定、灵敏度高,适合体外非那西丁及其代谢物对乙酰氨基酚的测定,可应用于体外CYP1A2酶活性的评价及酶动力学的研究.