牛膝中牛膝甾酮25位异构体构型的确定及其抗肿瘤活性的研究

目的牛膝 (AchyranthesbidentataBl.)中牛膝甾酮 2 5位异构体的分离、鉴定、构型确定以及肿瘤抑制活性的测定。方法利用硅胶吸附柱色谱和HPLC法进行分离 ,利用理化性质和波谱分析对化合物进行鉴定 ;利用体外实验法测定其活性。结果 2个非对映异构体化合物分别鉴定为 2 5 R牛膝甾酮 (2 5 Rinokosterone ,1)和 2 5 S牛膝甾酮 (2 5 Sinokosteone ,2 )。化合物 1具有一定的肿瘤抑制活性。结论 2个化合物均为首次从牛膝中分离得到 ,首次确定了其绝对构型 ,并首次对牛膝甾酮 2个 2 5位异构体进行了1H NMR、13 C NMR数据归属。牛膝中发挥肿瘤抑制作用的成分主要应为皂苷类成分。     

西洋参茎叶总皂苷氧化裂解产物中的新侧链环合型达玛烷型三萜

为了研究西洋参茎叶总皂苷氧化碱解产物的组成，对加拿大产西洋参茎叶总皂苷进行氧化碱解后，其产物通过硅胶柱色谱、Sephadex LH-20柱色谱、重结晶等方法进行分离纯化得到2个化合物。根据化合物的理化性质和光谱数据，鉴定其结构为：(12R,20S,24R)-20,24;12,24-diepoxy-24-deisopropyl-dammarane-3β-ol (1)和(20S,24R)-20,24-epoxydammarane-3β,12β,25-triol (2)。化合物1，2均为首次从西洋参茎叶总皂苷碱解产物中分离得到侧链环合的达玛烷型三萜，其中化合物1为新化合物。     

西洋参茎叶总皂苷碱降解成分西洋参茎叶总皂苷碱降解成分

目的研究西洋参茎叶总皂苷碱降解成分。方法采用硅胶柱色谱并结合HPLC进行分离纯化，通过波谱分析鉴定化合物的结构。结果从西洋参茎叶总皂苷碱降解产物中分离得到9种成分，分别鉴定为：20(S)-原人参二醇(I)，20(S)-达玛-25(26)-烯-3β,12β,20-三醇(II)，24(R)-ocotillol (III)，20(S)-原人参三醇(IV)，20(S)-达玛-25(26)-烯-3β,6α,12β,20-四醇(V)，达玛-20(21),24-二烯-3β,12β-二醇(VI)，达玛-20(21),24-二烯-3β,6α,12β-三醇(VII)，20(S),24(S)-达玛-25(26)-烯-3β,6α,12β,20,24-五醇(VIII)，20(S)-达玛-23-烯-25-过氧羟基-3β,6α,12β,20-四醇(IX)。结论碱降解20位S构型未改变。V，VII，VIII，IX为4个新化合物，并利用2D-NMR技术对新化合物的氢和碳的化学位移进行了归属。其中I对HCT-8人结肠癌细胞具有较强的细胞毒活性。     

HPLC法测定参乌益肾片中8种成分及其化学计量学综合评价

目的 建立HPLC法同时测定参乌益肾片中β-蜕皮甾酮、25R-牛膝甾酮、25S-牛膝甾酮、大车前苷、毛蕊花糖苷、异毛蕊花糖苷、木通苯乙醇苷B和2,3,5,4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷8种成分,联合化学计量学方法对参乌益肾片的产品质量进行综合评价。方法 采用Sepax Bio-C₁₈色谱柱（250 mm×4.6 mm,5 μm）;以乙腈–0.1%甲酸进行梯度洗脱;检测波长分别为250 nm（0～20 min测定β-蜕皮甾酮、25R-牛膝甾酮和25S-牛膝甾酮）、330 nm（20～60 min测定大车前苷、毛蕊花糖苷、异毛蕊花糖苷、木通苯乙醇苷B和2,3,5,4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷）;柱温30℃;体积流量1.0 mL/min;进样量10 μL。采用聚类分析、主成分分析和偏最小二乘法–判别分析法对8种指标成分检测结果相关性进行分析评价。结果 β-蜕皮甾酮、25R-牛膝甾酮、25S-牛膝甾酮、大车前苷、毛蕊花糖苷、异毛蕊花糖苷、木通苯乙醇苷B和2,3,5,4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷分别在1.47～36.75、0.66～16.50、0.74～18.50、5.80～145.00、15.77～394.25、4.58～114.50、1.29～32.25、8.78～219.50 μg/mL有良好的线性关系;平均回收率分别为97.78%、96.90%、97.96%、99.67%、100.01%、99.54%、98.03%、100.11%,RSD值分别为1.17%、0.82%、1.48%、0.93%、0.77%、0.69%、1.13%、0.65%。成分8（2,3,5,4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷VIP＝1.503）、成分4（大车前苷AVIP＝1.404）、成分5（毛蕊花糖苷VIP＝1.274）、成分1（β-蜕皮甾酮VIP＝1.030）是影响参乌益肾片样品质量的关键成分。结论 建立的方法简便快捷、准确可靠,可用于参乌益肾片中多指标成分的质量综合评价模式。     

衍生化法分离（2R，4R）-4-甲基-2-哌啶甲酸乙酯酒石酸盐手性异构体

目的 建立衍生化法分离（2R,4R）-4-甲基-2-哌啶甲酸乙酯酒石酸盐（MPFET）3种手性异构体。方法 以2,3,4,6-四-O-乙酰基-β-D-吡喃葡萄糖异硫氰酸酯（GITC）为柱前衍生化试剂,对MPFET手性异构体进行分离,并对衍生化条件进行优化。采用Venusil AQ C₁₈（250 mm×4.6 mm,5 μm）色谱柱作为固定相进行分离、监测和定量。以0.01 mol/L磷酸二氢钾溶液（用磷酸调pH至3.6）-乙腈（60∶40）为流动相,体积流量1.5 mL/min进行等度洗脱,采用紫外检测器,检测波长266 nm;柱温30℃;进样量20 μL。结果 MPFET与2S,4S-异构体分离度为1.76。2S,4R-异构体的线性范围为1.500～8.999 μg/mL,2R,4S-异构体的线性范围为0.255 2～1.531 0 μg/mL,2S,4S-异构体的线性范围为0.250 1～75.000 0 μg/mL,MPFET的线性范围为0.250 1～600.100 0 μg/mL,回收率均在90%～108%内,RSD均不大于3.0%。结论 柱前衍生化法分离MPFET中3种手性异构体,专属性强、准确度高、灵敏度高、重复性好,可用于MPFET手性异构体的分离和质量控制。     

构树叶的化学成分

为研究构树叶(Broussonetia papyrifera)的化学成分，用Diaion HP-20，Toyopearl HW-40C，Sephadex LH-20，silica gel等柱色谱方法进行分离，根据其理化性质和波谱数据鉴定化合物结构。分离得到了19个化合物，分别鉴定为芹菜素(1)，芹菜素-7-O-β-D-吡喃葡糖苷(2)，柯伊利素-7-O-β-D-吡喃葡糖苷(3)，芹菜素-7-O-β-D-吡喃葡糖醛酸苷(4)，牡荆素-7-O-β-D-吡喃葡糖苷(5)，木犀草素(6)，5,7,4′-三羟基-6-C-［a-L-鼠李糖(1→2)］-β-D-葡糖黄酮碳苷(7)，5,7,4′-三羟基-8-C-［α-L-鼠李糖(1→2)］-β-D-葡糖黄酮碳苷(8)，异牡荆素(9)，牡荆素(10)，苯甲酸苯甲酯-2,6-二-O-β-D-吡喃葡糖苷(11)，(2R,3R,5R,6S,9R)-3-羟基-5,6-环氧-β-紫罗兰醇-2-O-β-D-葡糖苷(12)，(2R,3R,5R,6S,9R)-3-羟基-5,6-环氧-乙酰-β-紫罗兰醇-2-O-β-D-葡糖苷(13)，ficustriol (14)，(6S,9S)-玫瑰花苷(15)，3β-羟基-5α,6α-环氧-β-紫罗兰酮-2α-O-β-D-葡糖苷(16)，icariside B₁ (17)，sammangaoside A (18)，3-羟基-5α,6α-环氧-β-紫罗兰酮(19)。化合物11、12、13为新化合物，其余化合物为首次从该属植物中分离得到。     

亮叶杨桐的三萜皂苷类成分

为研究亮叶杨桐(Adinandra nitida Merr. ex Li)叶的化学成分，亮叶杨桐叶的乙醇提取物经大孔树脂、硅胶、Sephadex LH-20以及ODS柱色谱分离得到6个化合物，通过波谱(¹H NMR、 ¹³C NMR、 HR-ESI-MS)和化学方法进行结构鉴定，6个化合物分别被鉴定为2α,3α,19α-trihydroxy-olean-12-en-28-oic acid-28-O-β-D-glucopyranoside (1)、 arjunetin (2)、 sericoside (3)、 glucosyl tormentate (4)、 nigaichigoside F1 (5)和arjunglucoside I (6)。化合物1为新化合物，化合物2～6均为首次从该植物中分离得到。     

多沙唑嗪对映体对兔四种血管α受体的作用

多沙唑嗪(doxazosin，DOX)作为高选择性α₁受体阻断药，是临床上治疗良性前列腺增生(benign prostatic hyperplasia, BPH)的一线药物，但同时引起心血管系统的不良反应。本研究采用兔离体动脉环张力实验及电场刺激兔离体隐动脉诱发交感嘌呤能血管收缩实验观察R-多沙唑嗪(R-doxazosin，R-DOX)和S-多沙唑嗪(S-doxazosin，S-DOX)对兔耳动脉、肠系膜动脉和肺动脉血管平滑肌α₁受体的作用，以及较高浓度R-DOX和S-DOX对兔隐动脉交感神经突触前膜α₂受体的作用。结果表明，在兔耳动脉、肠系膜动脉和肺动脉，R-DOX和S-DOX竞争性拮抗去甲肾上腺素(noradrenaline，NA)诱发的血管收缩反应；其pA₂值分别为7.91±0.03和7.53±0.05，7.80±0.05和7.29±0.07以及8.32±0.06和7.97±0.07；且S-DOX的pA₂值均明显小于R-DOX的pA₂值(P<0.01)。R-DOX和S-DOX(0.1～10 μmol·L^-1)对电刺激诱发的血管收缩反应无明显影响；R-DOX或S-DOX(100 μmol·L^-1)显著抑制电刺激诱发的血管收缩反应，完全抑制外源性NA诱发的血管收缩反应，但对1 mmol·L^-1腺苷三磷酸诱发的血管收缩反应无明显影响。上述结果提示，R-DOX和S-DOX对NA诱发兔耳动脉、肠系膜动脉和肺动脉收缩反应具有竞争性拮抗作用，对上述三种血管S-DOX拮抗NA的pA₂值均明显小于R-DOX。此外，R-DOX和S-DOX的浓度升至10 μmol·L^-1时，对血管交感神经末梢突触前膜α₂受体仍无明显影响。     

牛膝的化学成分

目的研究中药牛膝的化学成分。方法采用大孔树脂、硅胶柱色谱、制备型HPLC色谱等方法分离中药牛膝中的化学成分,依据这些化合物的理化性质和1H-NMR1、3C-NMR等波谱数据进行结构鉴定。结果从牛膝中分离鉴定了5个化合物,分别为:旌节花甾酮A(stachysterone A,1)、podecdysone C(2)、β-蜕皮甾酮(β-ecdysterone,3)、25-R-膝甾酮(25-R-inokosterone,4)、25-S-牛膝甾酮(25-S-inokosterone,5)。结论化合物1、2为首次从牛膝中分离得到。     

酞丁安对映体合成及其抗单纯疱疹病毒活性评价

酞丁安(3-酞酰亚胺-2-氧-正丁醛双缩氨硫脲,TDA)是药物研究所创制的抗病毒新药。为了研究其对映异构体(R),(S)-TDA对病毒活性及毒性是否有差异,并与消旋酞丁安(RS)-TDA的抗病毒活性及毒性进行比较,本文分别用已知构型的(R)-与(S)-丙氨酸为原料,通过缩合等6步反应,得到光学活性的(R)-,(S)-TDA,并与外消旋酞丁安比较其抗病毒活性及毒性。三者的抗单纯疱疹病毒活性与对细胞的毒性差别不大,说明消旋酞丁安临床使用是安全有效的。     

High performance liquid chromatography used for quality control of Achyranthis Radix

To establish a standard of quality control and to identify reliable Achyranthis Radix, three phytoecdysones including ecdysterone (1), 25R-inokosterone (2) and 25S-inokosterone (3) were determined by quantitative HPLC/UV analysis. Three phytoecdysones were separated with an YMC J'sphere ODS C(18) column (250 mm × 4.6 mm, 4 μm) by isocratic elution using 0.1% formic acid in water and acetonitrile (85:15, v/v%) as the mobile phase. The flow rate was 1.0 mL/min and the UV detector wavelength was set at 245 nm. The standards were quantified by HPLC/UV from Achyranthes bidentata Blume and Achyranthes japonica Nakai, as well as Cyathula capitata Moq. and Cyathula officinalis Kuan, which are of a different genus but are comparative herbs. The method was successfully used in the analysis of Achyranthis Radix of different geographical origin or genera with relatively simple conditions and procedures, and the assay results were satisfactory for linearity, recovery, precision, accuracy, stability and robustness. The HPLC analytical method for pattern recognition analysis was validated by repeated analysis of eighteen A. bidentata Blume samples and ten A. japonica Nakai samples. The results indicate that the established HPLC/UV method is suitable for quantitation and pattern recognition analyses for quality evaluation of Achyranthis Radix.     

板栗种仁化学成分的分离与鉴定

目的对板栗种仁的药用保健部位进行化学成分的分离与鉴定,为开发利用板栗种仁提供依据。方法板栗药用保健部位的正丁醇提取部位,经多次硅胶柱色谱和薄层色谱分离、理化常数测定、波谱分析、标准品比较等方法鉴定化合物的结构。结果分离得到6个化合物,分别为软脂酸-1-甘油单酯(hexadecanoic acid 2,3-dihydroxypropyl ester,1)、麦芽糖(maltose,2)、D-葡萄糖(D-glucose,3)、D-果糖(D-fructose,4)、5-羟甲基糠醛(5-hydroxymethylfurfural,5)、山柰酚(kaempferol,6)。结论化合物1、5为属内首次分离得到,化合物2、3、4、6为该种内首次分离得到。     

板栗种仁的化学成分

目的对板栗种仁的药用保健部位进行化学成分的研究。方法从板栗药用保健部位的乙酸乙酯提取部位经过多次硅胶柱色谱和薄层色谱分离化学成分,经理化常数测定、波谱分析、标准品比较和文献等鉴定出结构。结果分离得到7个化合物,分别为尿嘧啶(uracil,1)、正丁基-吡喃果糖苷(n-butyl-β-D-fructopyranoside,2)、壬二酸(azelaic acid,3)、异庚酸(isoenanthic acid,4)、蔗糖(su-crose,5)、β-谷甾醇(-βsitosterol,6)、胡萝卜苷(daucosterol,7)。其中化合物1-4为属内首次分离得到,化合物5为种内首次分离得到。     

A pair of 24-hydroperoxyl epimeric dammarane saponins from flower-buds of Panax ginseng

Further investigation on the saponins of the flower-buds of Panax ginseng C. A. Meyer has resulted in the isolation and structural elucidation of a pair of new 24-epimers of dammarane type saponins named ginsenoside I and II. The structures of the epimers were characterized on the basis of chemical and spectral evidence as 3-O-[β-D-glucopyranosyl-(1 → 2)-β-D-glucopyranosyl]-20-S-O-β-D-glucopyranosyl-3β12β20(S)-trihydroxy-24ξ-hydroperoxydammar-25-ene, except for their C-24 configurations. Ginsenoside I is a new triterpene glycoside, and ginsenoside II is a known compound first isolated from a natural plant.     

A Pair of 24-hydroperoxyl Epimeric Dammarane Saponins from Flower-buds of Panax Ginseng

Abstract

Further investigation on the saponins of the flower-buds of Panax ginseng C. A. Meyer has resulted in the isolation and structural elucidation of a pair of new 24-epimers of dammarane type saponins named ginsenoside I and II. The structures of the epimers were characterized on the basis of chemical and spectral evidence as 3-O-[β-D-glucopyranosyl-(1 → 2)-β-D-glucopyranosyl]-20-S-O-β-D-glucopyranosyl-3β12β20(S)-trihydroxy-24ξ-hydroperoxydammar-25-ene, except for their C-24 configurations. Ginsenoside I is a new triterpene glycoside, and ginsenoside II is a known compound first isolated from a natural plant.     

Gambogic acid and epigambogic acid, C-2 epimers with novel anticancer effects from Garcinia hanburyi

Gambogic acid, usually isolated as an inseparable stereomeric mixture of C-2 epimers, was newly separated into two epimers (1 and 2) from the gamboges of Garcinia hanburyi. The stereochemistry at C-2 was clearly defined by extensive spectroscopic analysis and direct comparison of NMR and HPLC data with those of the known R-epimer. Both epimers were examined for their cytotoxicities against human leukemia K562 (K562/S) and doxorubicin-resistant K562 (K562/R) cell lines. Different from doxorubicin (IC (50) = 10.78 microM for K562/R and 0.66 microM for K562/S), epimers 1 and 2 exhibited similar activities against both cell lines (IC(50) = 1.32 and 0.89 microM for 1, IC(50) = 1.11 and 0.86 microM for 2). These results suggested that both epimers were not multidrug resistance (MDR) substrates. Furthermore, epimers 1 and 2 were tested for their inhibitory effects against six human cytochrome P-450 enzymes. Epimers 1 and 2 showed little inhibitory effects toward five of the enzymes except CYP2C9. Interestingly, when tested against CYP2C9, S-epimer 2 had an inhibitory effect 20-fold stronger than that of R-epimer 1.     

Triterpene Saponins from the Roots of Achyranthes bidentata

Three saponins, oleanolic acid-28- O -ß-D-glucopyranoside (1), chikusetsusaponin V (2), and 3- O -ß-D-glucopyranosyl-oleanolic acid-28- O -ß-D-glucopyranoside (3) were isolated from the roots of Achyranthes bidentata Blume (Amaranthaceae). No activity was shown in the granulocyte phagocytosis test nor in the test of the potentiation of the cytotoxicity of cisplatin in human colon cancer cells. This is the first report of compounds 1, 2 and 3 isolated from Achyranthes species. Furthermore, the NMR data of 2 completed the partially published data.     

Two novel creatinine adducts of andrographolide in human urine

Andrographolide is a major labdane diterpenoid of the traditional Chinese and Ayurvedic medicine. Andrographis paniculate (Burm) Nees, is used in clinical situations in China mainly to treat fever, cold, and inflammation. In our previous study, fifteen metabolites of andrographolide were identified in human urine. However, there are still two other unknown metabolites. The aim of this study was to elucidate the structures of these two metabolites. 3. The two metabolites which are probably epimers were identified as creatinine adducts, and their structures were determined to be 14-deoxy-12-(creatinine-5-yl)-andrographolide-19-O-β-D-glucuronide A (Metabolite 1) and 14-deoxy-12-(creatinine-5-yl)-andrographolide-19-O-β-D-glucuronide B (Metabolite 2) by means of spectroscopic evidences. 4. It is for the first time that the formation of creatinine adducts as a novel metabolic pathway is reported. The mechanism was presumed that β-carbon (C-12) of α, β-unsaturated carbonyl was attacked by a 5-anion intermediate of creatinine formed through elimination of a proton, followed by the double bond migration from 12(13) to 13(14) and elimination of the hydroxyl group at C-14.