HPLC法同时测定泽泻中2种有效成分的含量

目的:建立泽泻药材中24-乙酰泽泻醇 A 和23-乙酰泽泻醇 B 含量的测定方法。方法:采用高效液相色谱法;色谱柱:大连依利特 Hypersil C_(18)柱(250 mm×4.6 mm,5 μm);检测器:蒸发光散射检测器(ELSD);柱温:室温;流动相:乙腈-水(75:25);流速:0.8 mL·min~(-1);ELSD 气体流速:2.00 L·min~(-1);漂移管温度:82℃。结果:被测定峰与其他组分峰可达到基线分离,泽泻中24-乙酰泽泻醇 A 和23-乙酰泽泻醇 B 的线性范围分别是0.33～1.98 μg(r=0.9992)及0.31～3.12 μg(r=0.9999);平均回收率(n=6)分别为100.8%及98.7%,RSD 分别为1.2%及3.3%。结论:该方法准确、简便、重复性好,可作为泽泻中24-乙酰泽泻醇 A 和23-乙酰泽泻醇 B 的含量测定方法。     

反相高效液相色谱法测定泽泻提取物中23-乙酰泽泻醇B含量

目的 利用反相高效液相色谱法(RP-HPLC)测定泽泻提取物中23-乙酰泽泻醇B含量.方法 利用RP-HPLC检测,选用Kromasil 100-5C18(250 mm×4.6 mm)色谱柱,以乙腈-水为流动相等梯度洗脱,流速1ml·min-1;检测波长208 nm.结果 23-乙酰泽泻醇B在进样量范围内与峰面积线性关系良好,仪器精密度和稳定性RSD均小于2％,平均回收率在99％～101％(n=3).测定了5批提取物中23-乙酰泽泻酐B,平均的含量为2.550 mg/g.结论 该方法快速、准确、重复性好,可对泽泻指标性成分23-乙酰泽泻醇B进行定量,从而控制提取物质量.     

HPLC法测定痛风定胶囊中龙胆苦苷、马钱苷酸、獐牙菜苦苷、獐牙菜苷、杯苋甾酮、泽泻醇F、泽泻醇A、24-乙酰泽泻醇A、23-乙酰泽泻醇B

目的 建立高效液相色谱法同时测定痛风定胶囊中龙胆苦苷、马钱苷酸、獐牙菜苦苷、獐牙菜苷、杯苋甾酮、泽泻醇F、泽泻醇A、24-乙酰泽泻醇A和23-乙酰泽泻醇B.方法 采用HPLC法,Waters Symmetry C18色谱柱(250 mm×4.6 mm,5μm);流动相:乙腈–0.1％磷酸水溶液,梯度洗脱;检测波长:24...     

Syntheses of 2, 5-dialkylfuran and tetrahydrofuran carbinols and their cytotoxic activity

2, 5-Dialkylfuran and tetrahydrofuran compounds as structural elements of Annonaceae acetogenins like Asitrocinwere synthesized starting from furfural by Grignard reactions, lithiation of the furan ring and addition of aliphatic aldehydes. Hydrogenation of the furan rings over Pd-catalyst gave the corresponding tetrahydrofurans. All resulting compounds showed no or rather less antimicrobial activity against grampositive, gram-negative bacteria and fungi compared to tetracycline or clotrimazol but high cytotoxic activity against HL 60 cell line determined in the MTT assay.     

23-乙酰泽泻醇B对2型糖尿病小鼠血糖的影响

目的探讨23-乙酰泽泻醇B是否有治疗2型糖尿病的潜能。方法通过腹腔注射链脲佐菌素和烟酰胺建立2型糖尿病小鼠模型。灌胃罗格列酮或23-乙酰泽泻醇B 3周后,测定2型糖尿病小鼠血糖值,次日进行口服葡萄糖耐受试验(OGTT)。采用葡萄糖荧光示踪剂,测定23-乙酰泽泻醇B对葡萄糖吸收的影响。采用3T3-L1前脂肪细胞分化模型,测定其对分化的影响。结果分别每天灌胃阳性药罗格列酮10 mg·kg~(-1)、23-乙酰泽泻醇B(5、10、20 mg·kg~(-1)),连续灌胃给药3周后,降低了2型糖尿病小鼠血糖值,一定程度改善OGTT过程中胰岛素抵抗。在30 mmol·L~(-1)高糖条件下,23-乙酰泽泻醇B促进了脂肪细胞对胰岛素刺激的葡萄糖吸收;23-乙酰泽泻醇B(1、10μmol·L~(-1))促进3T3-L1前脂肪细胞的分化过程。结论 23-乙酰泽泻醇B降低2型糖尿病小鼠血糖,促进前脂肪细胞分化,促进脂肪细胞吸收葡萄糖,但作用机制仍需进一步探索。     

Synthesis and cytotoxic activity of 1,3-benzodioxole derivatives. Note II

A series of 1,3-benzodioxoles (2-12) were synthesized and evaluated for their in vitro ability to inhibit the growth of three human tumor cell lines. No cytotoxic effects were noticed with any of the test compounds at a concentration of 10(-4) M.     

Chemical constituents of the leaves of Diospyros kaki and their cytotoxic effects

Isolation and structure elucidation of two new compounds, kakispyrone (1) and kakisaponin A (2), together with 11 known compounds, from the leaves of Diospyros kaki L. are described. Their cytotoxic effects against several cancer cell lines (A549, HepG2 and HT29) are also reported.     

A UFLC/MS/MS method for simultaneous quantitation of alisol A and alisol B 23-acetate from Alisma orientale (Sam.) Juz. in rat plasma

A sensitive and reliable ultra fast liquid chromatography tandem mass spectrometry (UFLC-MS/MS) method has been developed and validated for simultaneous quantitation of alisol A and alisol B 23-acetate from Alisma orientale (Sam.) Juz. in rat plasma using diazepam as an internal standard (IS). The plasma samples were extracted by liquid–liquid extraction with methyl tert-butyl ether and separated on a Venusil MP C18 column (100 mm × 2.1 mm, 3.0 mm) (Venusil, China) using gradient elution with the mobile phase consisting of methanol and 0.1% acetic acid in water at a flow rate of 0.4 ml/min. The two analytes were monitored with positive electrospray ionization by multiple reaction monitoring mode (MRM). The lower limit of quantitation was 5.00 ng/ml for alisol A and 5.00 ng/ml for alisol B 23-acetate. The calibration curves were linear in the range of 5.00–2500 ng/ml for alisol A and 5–2500 ng/ml for alisol B 23-acetate. The mean extraction recoveries were above 63.8% for alisol A and 68.0% for alisol B 23-acetate from biological matrixes. Both intra-day and inter-day precision and accuracy of analytes were well within acceptance criteria (15%). The validated method was successfully applied to the pharmacokinetic study of alisol A and alisol B 23-acetate in rat plasma after oral administration of alcohol extract of Alismatis Rhizoma.     

CIK细胞杀瘤实验前后细胞表型与细胞毒性相关性研究

目的为了分析体外细胞因子诱导培养CIK细胞在体外杀瘤实验中的细胞表型变化与其杀瘤活性的相关性及为临床过继免疫治疗提供实验依据。方法采用体外诱导方法扩增培养正常人外周血淋巴细胞及单个核细胞,用CCK-8试剂检测培养14d的CIK细胞对SPC-A-1和BGC-823肿瘤细胞的细胞毒活性,应用流式细胞术测定杀瘤实验前后CD3＋等6种不同表型细胞百分率的变化。结果在与效靶比成正比的条件下,体外实验CIK细胞对两种肿瘤细胞的杀伤率都与CD3＋CD4＋,CD3＋CD4-CD8-细胞比例变化呈正相关（P〈0.05）,都与CD3＋CD25＋,CD3＋CD28＋,CD3＋CD25＋CD28＋细胞比例变化呈负相关（P〈0.05）,对SPC-A-1细胞的杀瘤率与CD3＋CD16＋CD56＋呈正相关（P〈0.05）。结论本研究表明靶细胞抗原在活化细胞中起重要作用,测定培养细胞活化相关表型可以间接监测其杀瘤能力,为临床CIK细胞过继免疫治疗的应用提供实验依据。     

高效液相色谱法测定五苓滴丸中23-乙酰泽泻醇B的含量

目的采用高效液相色谱(HPLC)法测定五苓滴丸中23-乙酰泽泻醇B的含量。方法色谱柱:Agilent Eclipse XDB-C18柱(4.6 mm×250 mm,5μm);流动相:乙腈-水(70∶30);流速:1.0 mL/min;检测波长210 nm;柱温:30℃。结果 23-乙酰泽泻醇B在2.9~145.0μg/mL浓度范围内与峰面积具有良好的线性关系(r=0.999 9),平均加样回收率为97.86%,RSD=1.29%。结论 HPLC法简便、准确、专属性高,可用于五苓滴丸的质量控制。     

基于荧光图像的抗肿瘤细胞迁移药物筛选方法

本文建立了一种基于荧光图像的抗肿瘤细胞迁移药物筛选方法。应用Hoechst 33342荧光染料染色和96孔Transwell板,通过细胞荧光图像采集与分析平台对正常组和加药组细胞荧光图像进行定量分析,建立了一种快速筛选抗肿瘤细胞迁移药物的新方法。对本方法的稳定性、精密度、荧光染色时间和迁移时间进行了考察和优化,并对抗肿瘤药紫杉醇的肿瘤细胞迁移抑制作用进行了定量分析,计算得到紫杉醇抗肿瘤细胞迁移的IC50为0.717μmol.L-1。采用该方法对中药泽泻24个组分进行筛选,发现1个活性组分。结果表明,该方法快速、准确、重现性好,能够用于抗肿瘤细胞迁移药物的快速筛选。     

Chemical constituents of the Annona glabra fruit and their cytotoxic activity

Abstract

Context: Traditional Chinese medicines have attracted increasing interest as potential sources of novel drugs with a wide range of biological and pharmacological activities. Annona glabra Linn (Annonaceae) is used in traditional medicine as an anticancer drug. Phytochemical investigation of this plant led to the isolation of acetogenins, ent-kauranes, peptides, and alkaloids. In addition, compounds exhibited anticancer, anti-HIV-reserve, and antimalaria.

Objective: Isolation, structure determination, and cytotoxic activity evaluation of compounds from the methanol extract from A. glabra fruits.

Materials and methods: Using chromatographic methods to isolate compounds from the A. glabra methanol extract. The cytotoxic activity of compounds was evaluated by a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. In addition, compounds which showed significant cytotoxic activity were chosen for further study apoptosis characteristics.

Results: One new, (2E,4E,1′R,3′S,5′R,6′S)-dihydrophaseic acid 1,3′-di-O-β-d-glucopyranoside, and eight known compounds, (2E,4E,1′R,3′S,5′R,6′S)-dihydrophaseic acid 3′-O-β-d-glucopyranoside (2), icariside D₂ (3), icariside D₂ 6′-O-β-d-xylopyranoside (4), 3,4-dimethoxyphenyl O-β-d-glucopyranoside (5), 3,4-dihydroxybenzoic acid (6), blumenol A (7), cucumegastigmane I (8), and icariside B₁ (9), were isolated from the fruits of A. glabra. Icariside D₂ (3) was found to show significant cytotoxic activity on the HL-60 cell line with the IC₅₀ value of 9.0?±?1.0?µM and did not show cytotoxic activity on the Hel-299 normal cell line. The further test indicated that compound 3 induced apoptosis via alteration of expression of apoptosis-related proteins and decreased phosphorylation of AKT in HL-60 cells.

Discussion and conclusion: The results suggested that the constituents from A. glabra may contain effective compounds which can be used as anticancer agents.     

Synthesis and cytotoxic activities of 8-alkyl or 8-aryl-8,9-dihydro-7H-isoindolo[5,6-g]quinoxaline-7,9-diones

A series of 8-alkyl- and 8-aryl-8,9-dihydro-7H-isoindolo[5,6- g]quinoxaline-7,9-diones were synthesized using sultine chemistry as a key step in good yield. These compounds were evaluated for their in vitro cytotoxicity against six human cancer cell lines (HCT15, SK-OV-3, A549, SNB19, MCF7 and MCF7/ADR).     

Antioxidant, cytotoxic activities, and structure-activity relationship of gallic acid-based indole derivatives

A new series of gallic hydrazones containing an indole moiety was synthesized through the reaction of gallic hydrazide and different indole carboxaldehydes. Their antioxidant activities were determined on DPPH radical scavenging and inhibition of lipid peroxidation. The in-vitro cytotoxic activities of the compounds were evaluated against HCT-116 (human colon cancer cell line) and MCF-7 (estrogen-dependent human breast cancer cell line) by the MTT method. An attempt to correlate the biological results with their structural characteristics has been done. A limited positive structure activity relationship was found between cytotoxic and antioxidant activities.     

Stilbenes and oligostilbenes from leaf and stem of <Emphasis Type="Italic">Vitis amurensis</Emphasis> and their cytotoxic activity

Chromatographic separation of the EtOAc fraction from the leaf and stem of Vitis amurensis led to the isolation of six oligostilbenoids (i.e., r-2-viniferin (1), trans-amurensin B (2), trans-ɛ-viniferin (3), gnetin H (4), amurensin G (5), (+)-ampelopsin A (8)) and four stilbenoids (i.e., trans-resveratrol (6), (+)-ampelopsin F (7), piceatannol (9), and trans-piceid (10)). The structures have been identified on the basis of spectroscopic evidence and physicochemical properties. The isolates were investigated for cytotoxic activity against three cancer cell lines in vitro using the MTT assay method. Amurensin G (5) and trans-resveratrol (6) showed significant cytotoxic activity against L1210, K562 and HTC116 cancer cell lines with IC₅₀ values ranging from 15.7 ± 2.1 to 30.9 ± 1.8 μM. (+)-Ampelopsin A (8) and trans-piceid (10) exhibited considerable cytotoxic activity against L1210 (IC₅₀ values of 30.6 ± 4.1 and 28.7 ± 2.81 μM, respectively) and K562 (IC₅₀ values of 38.6 ± 0.82 and 24.6 ± 0.76 μM, respectively). Gnetin H (4) showed only weak cytotoxic activity against L1210 with an IC₅₀ value of 40.1 ± 4.23 μM. On the other hand, r-2-viniverin (1), trans-amurensin B (2), trans-ɛ-viniferin (3), (+)-ampelopsin F (7), and piceatannol (9) exhibited no activity on three cancer cell lines.     

Synthesis of N-(phenylcarbamothioyl)-benzamide derivatives and their cytotoxic activity against MCF-7 cells

Cancer is one of the leading causes of death both in developing countries and across the globe. In Indonesia, cancer ranks as the fifth primary cause of death following heart disease, stroke, respiratory tract and diarrhea. Therefore, studies on thiourea derivative compounds as anticancer agents have been profoundly conducted but still require further continuous development. In the present study, we aimed tosynthesize new anticancer compounds of N-(phenylcarbamothioyl)-benzamide derivatives, namely N-(phenylcarbamothioyl)-4-bromobenzamide and N-(phenylcarbamothioyl)-4-fluorobenzamide compounds and assess their activities against MCF-7 breast cancer cells. The initial step was to predict the drug-receptor activity through docking between the tested compounds using epidermal growth factor receptor (EGFR) (PDB code: 1M17). The compounds were futher synthesized from the reactions between benzoyl chloride derivatives and N-phenylthiourea. The structures of the new compounds were identifiedusing FTIR, ¹H NMR, ¹³C NMR and mass spectra. The cytotoxic activities (IC₅₀) to breast cancer cells of MCF-7 N-(phenylcarbamothioyl)-4-bromobenzamide compound and N-(phenylcarbamothioyl)-4-fluorobenzamide were 0.27 mM and 0.31 mM, respectively. These two new compounds had better cytotoxic activities than those of the currenthydroxyurea-based anticancer drugs (the reference compound) with an IC₅₀ value of 9.76 mM. Furthermore, these two newcompounds were not toxic to Vero normal cells. Therefore, they possessedtremendous potentials as the candidates for new drugs against breastcancer.     

Chemical constituents from the roots of Tripterygium wilfordii and their cytotoxic activity

In our ongoing search for bioactive constituents, a new sesquiterpene polyol ester, named triptersinine U (1), together with five known triterpenes (2–6) and seven sesquiterpene pyridine alkaloids (7–13), were isolated from the roots of Tripterygium wilfordii Hook. f. Their chemical structures were elucidated using extensive spectroscopic analyses, including 1D and 2D NMR, and HRESIMS, as well as comparison with previously reported data. Cytotoxic activities of all compounds 1–13 were evaluated against six human tumor cell lines (HepG2, Hep3B, Bcap37, U251, MCF-7 and A549) using the MTT in vitro assay. The results showed that triterpenes exhibited moderate cytotoxic activities toward the tested cell lines.     

Reversal of P-glycoprotein-mediated multidrug resistance by Alisol B 23-acetate

Herbal drugs were screened for their activity in reversing multidrug resistance (MDR) in P-glycoprotein (P-gp) over-expressing cancer cells. Through bio-assay guided fractionation an active compound was isolated from Rhizoma Alismatis, the underground part of Alisma orientale and the chemical structure of the isolate compound was confirmed by HPLC, LC-MS and NMR as Alisol B 23-acetate (ABA). ABA restored the sensitivity of MDR cell lines HepG2-DR and K562-DR to anti-tumor agents that have different modes of action but are all P-gp substrates. It restored the activity of vinblastine, a P-gp substrate, in causing G2/M arrest in MDR cells. In a dose-dependent manner, ABA increased doxorubicin accumulation and slowed down the efflux of rhodamin-123 from MDR cells. ABA inhibited the photoaffinity labeling of P-gp by [125I]iodoarylazidoprazosin and stimulated the ATPase activity of P-gp in a concentration-dependent manner, suggesting that it could be a transporter substrate for P-gp. In addition, ABA was also a partial non-competitive inhibitor of P-gp when verapamil was used as a substrate. Our results suggest that ABA may be a potential MDR reversal agent and could serve as a lead compound in the development of novel drugs.     

Evaluation of cytotoxic and apoptotic effects of the extracts and phenolic compounds of Astragalus globosus Vahl and Astragalus breviflorus DC

Astragalus L. is a genus member of the Fabaceae family, representing about 3,000 species all over the world and 380 species in Turkey. Astragalus species have been used in traditional medicine for many years. Astragalus globosus Vahl, known as “top geven”, is a dwarf, scapose, perennial herb, Astragalus breviflorus DC., known as “yünlü geven”, is an extremely spiny dwarf shrub. These endemic species grow in the Turkish cities of Erzurum, Kars, and Van. This is the first phytochemical and cytotoxic investigation of Astragalus globosus Vahl and Astragalus breviflorus DC. The main extracts and sub-fractions from the plants were evaluated for in vitro cytotoxic and apoptotic activities. The IC₅₀ values of dichloromethane, n-butanol, and water extracts of the aerial parts of A. globosus against the MCF-7 cell line were determined as 28.39, 868.60, and 1753.00 µg/mL. The values for the MDA-MB-231 cell line were 264.00, 620.30, and 1300.50 µg/mL, respectively. From A. globosus, the following were isolated: a flavone glycoside, diosmetin-7-O-rutinoside (1); and two flavonol glycosides, isorhamnetin-3-O-rutinoside (2) and quercetin-3-O-galactoside (3). From A. breviflorus, two phenolic acids, caffeic acid (4) and chlorogenic acid (5), and a flavan-3-ol, catechin (6), were isolated. Diosmetin-7-O-rutinoside was isolated from Astragalus species for the first time and showed the highest cytotoxic activities on the MCF-7 and MDA-MB-231 breast cancer cell lines with IC₅₀ values of 13.65 and 12.89 μg/mL, respectively. Moreover, we observed that diosmin exerts cytotoxic effects by causing cell necrosis.     

Six new furostanol glycosides from Smilax glauco-china and their cytotoxic activity

Six new steroidal saponins, namely glauco-chinaosides A–F, and one known compound were isolated from the tubers of Smilax glauco-china. Their structures were elucidated by a combination of spectroscopic analysis and hydrolysis followed by spectral and chromatographic analysis. Compounds 1–7 were tested in vitro for their cytotoxic activities against four human tumor cell lines (SH-SY5Y, SGC-7901, HCT-116, and Lovo). Compounds 1, 2, and 5 exhibited cytotoxic activity against SGC-7901, with IC₅₀ values of 2.7, 11.5, and 6.8 μM, respectively.