The cytotoxicity and mechanisms of 1,2-naphthoquinone thiosemicarbazone and its metal derivatives against MCF-7 human breast cancer cells

We have investigated the antitumor functions and mechanisms of 1,2-naphthoquinone-2-thiosemicarbazone (NQTS) and its metal complexes (Cu(2+), Pd(2+), and Ni(2+)) against MCF-7 human breast cancer cells. The cells were dosed with these complexes at varying concentrations, and cell viability was measured by a sulforhodamine B (SRB) method. To study mechanisms of action, the complexes were incubated with topoisomerase II (topo II) and supercoiled DNA, linear DNA, nicked open DNA, and relaxed DNA were detected by agarose gel electrophoresis. The results revealed that these complexes are effective antitumor chemicals in inhibiting MCF-7 cell growth, with Ni-NQTS being the most effective among the complexes studied. Our data also indicated that Ni-NQTS is more effective than the commercial antitumor drug, etoposide, based on IC(50) values. The mechanistic study of action showed that metal complexes of NQTS, NQ, and NQTS can only stabilize the single-strand nicked DNA, but not double-strand breakage intermediates. In addition, metal derivatives of these ligands, but not the parent NQ and NQTS, exerted an antagonizing effect on topoisomerase II activity. In summary, chemicals with or without metal derivatives might possess different chemical-topoisomerase II-DNA interactions.     

咖啡酸、阿魏酸和香草酸对酪氨酸酶活性的影响

目的 :研究咖啡酸、阿魏酸和香草酸对酪氨酸酶活性的影响 ,寻找酪氨酸酶抑制剂 ,从而为治疗色素增加性皮肤病筛选药物。方法 :酪氨酸酶多巴速率氧化法体外测定药物干预前后酪氨酸酶活性 ,求出酪氨酸酶抑制率。采用Lineweaver Burk双倒数法制得酶动力学曲线 ,推断抑制类型。结果 :3种试药中仅香草酸对酪氨酸酶具有抑制作用 ,与氢醌比较无显著差异 (P >0 .0 5 ) ,咖啡酸、阿魏酸对酪氨酸酶具有上调激活作用。结论 :香草酸为良好的混合型酪氨酸酶抑制剂。     

New gastroprotective ferruginol derivatives with selective cytotoxicity against gastric cancer cells

The diterpene ferruginol has shown a strong protective effect in animal gastric ulcer models. In the present work, we report the gastroprotective effect and cytotoxicity of 16 new semisynthetic ester derivatives of ferruginol. The gastroprotective effect of these compounds was assessed with the HCl/EtOH-induced gastric lesions model in mice and the cytotoxicity was measured using MRC-5 fibroblasts, gastric adenocarcinoma (AGS) and liver hepatoma Hep G2 cells. The compounds were tested for a gastroprotective effect at a single oral dose of 20 mg/kg. The best gastroprotective effect was elicited by ferruginyl nicotinate ( 13), reducing the lesion index by 71 %, while the derivatives ferruginyl chloroacetate ( 2), ferruginyl palmitate ( 6), ferruginyl oleate ( 7), ferruginyl 3,5-dinitrobenzoate ( 11), ferruginyl 3-methylbenzofuran-2-carbonyl ester ( 12), ferruginyl indoleacetate ( 14), ferruginyl indolebutyrate ( 15) and ferruginyl pthalate ( 16) reduced the lesions by 49 - 66 %. The most promising compounds were 11, 13 and 14, presenting a gastroprotective effect higher or similar to that of ferruginol but with a high selectivity towards the tumor AGS cells. Among the three products, the most selective towards AGS cells was 14, followed by 13, and 11 (IC (50) values of 12, 22 and 29 microM, respectively). The isobutyrate 4, inactive as a gastroprotective agent, showed selective cytotoxicity against AGS and Hep G2 cells (IC (50) values of 60 and 39.2 microM, respectively). The cytotoxicity of the above cited compounds towards fibroblasts was >1000 microM. Considering the aliphatic esters of ferruginol, the best gastroprotective activity was found in the C (16) and C (18) derivatives but tended to decrease with increasing aliphatic chain unsaturation. For short-chain esters, the gastroprotective effect could be observed when the chain contained a chlorine atom. For aromatic esters, the presence of nitro groups or a nitrogen atom in the aromatic ring enhanced the gastroprotective activity. The compounds with the best gastroprotective effect and the highest selectivity against tumor cells bear an amino group (indoleacetate and nicotinate) or nitro group (3,5-dinitrobenzoate).     

高效液相色谱法同时测定蒲公英中咖啡酸和阿魏酸的含量

目的建立了RP-HPLC法对蒲公英中咖啡酸和阿魏酸同时定量,考察不同产地蒲公英中咖啡酸、阿魏酸的含量。方法利用高效液相色谱法梯度洗脱。色谱条件为:Hypersil BDS C18分析柱(5μm,4.6mm×250 mm),柱温40℃,流动相为甲醇?0.01M磷酸二氢钾溶液(磷酸调pH为3.7)梯度洗脱,流速1.0mL.m in-1,检测波长323nm。结果咖啡酸、阿魏酸浓度与峰面积呈良好线性关系,线性范围分别是:咖啡酸8.192~81.92μg.mL-1,r=0.9991(n=6);阿魏酸1.96~39.2μg.mL-1,r=0.9992(n=6);回收率咖啡酸为95.3%~98.2%,阿魏酸为92.1%~95.3%(n=9)。结论本方法测定了9个不同产地或不同批号的蒲公英样品中咖啡酸、阿魏酸的含量,该方法分离度好,快速,简便,重现性好。     

HPLC法同时测定升麻甘草汤中咖啡酸和阿魏酸的含量

目的建立升麻甘草汤的质量标准控制方法.方法色谱柱DiamonsilC18(250mm×4.6mm,5μm),流动相:甲醇-0.1 mo·lL-1醋酸铵-冰醋酸(20∶80∶1);流速:1.0mL·min-1;检测波长:310 nm.结果咖啡酸在0.2～4mg·L-1浓度范围内呈良好的线性关系,平均回收率为103.6%(n=9);阿魏酸在1～10mg·L-1浓度范围内呈良好的线性关系,平均回收率为104.3%(n=9).结论本法简便、灵敏、准确,可有效控制升麻甘草汤的质量.     

HPLC法同时测定活血止痛液中阿魏酸和咖啡酸的含量

目的：建立同时测定活血止痛液中阿魏酸和咖啡酸含量的HPLC方法。方法：采用Inertsil ODS-SP C18色谱柱（4.6 mm×250 mm,5μm）,流动相为乙腈-0.05%磷酸溶液（17∶83）,流速为1.0 mL· min-1,柱温为30℃,检测波长为320 nm,进样量为10μL。结果：阿魏酸和咖啡酸与其他杂质峰分离良好。阿魏酸在1.766~35.328μg· mL-1范围内线性关系良好（r =0.9999）;咖啡酸在0.835~16.704μg· mL-1范围内呈现良好的线性关系（r=0.9993）,阿魏酸和咖啡酸的平均加样回收率分别为100.79%、101.05%,RSD分别为1.73%（n=9）、1.87%（n=9）。结论：本方法简便,测定结果准确,重复性好,可作为活血止痛液的质量控制方法。     

Effects of selected flavonoids and caffeic acid derivatives on hypoxanthine-xanthine oxidase-induced toxicity in cultivated human cells

Tissue damage as a result of oxygen radical generation may be involved in the pathogenesis of different diseases, carcinogenesis, aging and cell death. The inhibition of the proliferation rate of the immortalised human cell line ECV 304 after oxidant damage by oxygen radicals generated in a hypoxanthine-xanthine oxidase system and the protection provided by some selected flavone and flavonol glycosides as well as by caffeic acid and its derivatives was determined. The cytotoxicity of the reactive oxygen species was differentially influenced by selected flavonoids and seems to be determined by the pattern of substitution and by their lipophilicity. Apigenin and quercetin demonstrated the strongest effect on the inhibition of hypoxanthine-xanthine oxidase-induced toxicity (50 % restitution of the cells at a concentration of 0.36 microM and 3.1 microM, respectively). The beneficial effect of the flavonol glycosides rutin and hyperoside was weak, whereas flavone glycosides such as diosmin showed a better effect of protection.     

Abemaciclib-loaded ethylcellulose based nanosponges for sustained cytotoxicity against MCF-7 and MDA-MB-231 human breast cancer cells lines

Abemaciclib (AC) is a novel, orally available drug molecule approved for the treatment of breast cancer. Due to its low bioavailability, its administration frequency is two to three times a day that can decrease patient compliance. Sustained release formulation are needed for prolong the action and to reduce the adverse effects. The aim of current study was to develop sustained release NSs of AC. Nanosponges (NSs) was prepared by emulsion-solvent diffusion method using ethyl-cellulose (EC) and Kolliphor P-188 (KP-188) as sustained-release polymer and surfactant, respectively. Effects of varying surfactant concentration and drug: polymer proportions on the particle size (PS), polydispersity index (PDI), zeta potential (ζP), entrapment efficiency (%EE), and drug loading (%DL) were investigated. The results of AC loaded NSs (ACN1-ACN5) exhibited PS (366.3–842.2 nm), PDI (0.448–0.853), ζP (−8.21 to −19.7 mV), %EE (48.45–79.36%) and %DL (7.69–19.17%), respectively. Moreover, ACN2 showed sustained release of Abemaciclib (77.12 ± 2.54%) in 24 h Higuchi matrix as best fit kinetics model. MTT assay signified ACN2 as potentials cytotoxic nanocarrier against MCF-7 and MDA-MB-231 human breast cancer cells. Further, ACN2 displayed drug release property without variation in the % release after exposing the product at 25 °C, 5 °C, and 45 °C storage conditions for six months. This investigation proved that the developed NSs would be an efficient carrier to sustain the release of AC in order to improve efficacy against breast cancer.     

绿原酸、咖啡酸、阿魏酸对微血管内皮细胞的损伤作用

<正>微血管内皮细胞具有重要的生理功能~([1])。诸多因素会损伤微血管内皮细胞,从而导致微血管内皮功能的失衡,进而引起多种病征和损伤~([2])。其中,药物损伤是一个重要的因素。绿原酸(chlorogenic acid,CGA)是金银花、杜仲等药材中的重要活性成分,也是诸多中药民族药的质量控制的主要指标之一,同时也是具有重要应用开发前景的活性物质~([3])。     

Synthesis and anticancer evaluation of N-benzoyl-N'-phenyltiourea derivatives against human breast cancer cells (T47D)

New anti-breast cancer compounds have been found and may prove to have stronger activity. To predict the activities of N-benzoyl-N'-phenylthiourea (BPTU) derivatives, namely N-(3-chloro)benzoyl-N'-phenylthiourea (3-Cl-BPTU) and N-(3,4-dichloro)benzoyl-N'-phenylthiourea (3,4-2Cl-BPTU) with Sirtuin-1 receptor (PDB code: 4I5I), molecular docking was conducted at the beginning of this study. The compounds were then synthesized from benzoyl chloride derivatives and N-phenylthiourea. Molecular structure was confirmed using FTIR, ¹H NMR, ¹³C NMR and Mass Spectra, while the anticancer activity was tested in vitro against human breast cancer cells (T47D) using MTT assay. The results indicated that the anti-cancer activities of the test compounds were better than those of the hydroxyurea as the reference compound, evidenced by the Rerank Score (RS). Furthermore, cytotoxic effect of 3-Cl-BPTU (IC₅₀: 0.43 mM) and 3,4-dichloro-BPTU (IC₅₀: 0.85 mM) showed better result compared with hydroxyurea (IC₅₀: 4.58 mM). Therefore, we concluded that these compounds could possess termendous potential as the candidate for a new anticancer drug.     

RP-HPLC法同时测定升麻中3种有机酸的含量

目的建立同时测定升麻中咖啡酸、阿魏酸和异阿魏酸含量的方法。方法采用RP-HPLC法,C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-水-磷酸(13.00∶87.00∶0.05),检测波长为320 nm。结果咖啡酸、阿魏酸和异阿魏酸的质量浓度分别在2.66~26.60 mg.L-1(r=0.999 5)、6~60 mg.L-1(r=0.999 3)和20.6~206.0 mg.L-1(r=0.999 3)内与峰面积呈良好的线性关系(n=6),方法回收率(n=9)分别为99.8%、98.2%和101.0%。结论RP-HPLC法可用于升麻中3种有机酸含量的同时测定。     

In vitro antimetastatic activity of koetjapic acid against breast cancer cells

Breast cancer is the most common cancer in women, and it can metastasize very rapidly. Tumor metastasis is the primary cause of cancer deaths. In the present study, we investigated the capability of koetjapic acid, a natural triterpene, in the induction of apoptosis and the inhibition of metastasis in the breast cancer cell line (MCF 7). The effects of koetjapic acid against 4 steps of metastasis have been assessed, including cell survival, clonogenicity, migration and invasion. Koetjapic acid exhibited cytotoxic activity against MCF 7 cells with an IC(50) of 68.88±6.075 μg/mL. The mechanism of cell death was confirmed due to the induction of apoptosis machineries; early and late apoptosis-related changes were detected, including the stimulation of caspase 3/7 activities, apoptosis-related morphological changes such as membrane blebbing, chromatin condensation and DNA fragmentation. A mitochondrial apoptosis pathway was found to be involved in koetjapic acid-induced cell death induction. Moreover, at a sub-toxic dose (15 μg/mL), Koetjapic acid inhibited cell migration and invasion significantly. Finally, koetjapic acid inhibited the colony formation properties of MCF 7 significantly. These results indicate that koetjapic acid possesses significant antitumor and antimetastatic effects, and warrants further investigation.     

盐霉素和盐霉素钠对人乳腺癌干细胞和乳腺癌细胞体外细胞毒作用的比较(英文)

盐霉素是一种从白色链丝菌培养物中分离提取的聚醚类离子载体型抗生素,它广泛应用于畜禽类动物鸡球虫病的防治,并且还能作为饲料添加剂以促进畜禽的生长。最近新发现盐霉素具有特异性抑制肿瘤干细胞的作用。目前盐霉素钠的价格只是盐霉素的十分之一,但两者在抑制肿瘤细胞和肿瘤干细胞方面的异同却未见文献报导。本研究以人乳腺癌细胞MCF-7及其干细胞为模型,通过SRB实验对盐霉素及其钠盐的细胞毒性进行了评价和比较。首先,通过流式细胞仪从MCF-7细胞中分选得到了SP细胞;然后用乳腺癌干细胞表面特异性标志物CD44~+/CD24~-对SP细胞进行了鉴定;最后,分别测定了盐霉素和盐霉素钠对分选得到的肿瘤干细胞和肿瘤细胞的体外生长抑制率。结果表明,与乳腺癌细胞相比较,盐霉素和盐霉素钠对肿瘤干细胞均表现出了更强的抑制作用,在同样的给药浓度下,盐霉素和盐霉素钠对肿瘤细胞和肿瘤干细胞的抑制作用未表现出明显的差异。本研究结果提示,在抑制肿瘤干细胞的相关研究中,可以用盐霉素钠替代盐霉素而不会影响抑制效果。     

基于UPLC法同时测定三叉神经散中绿原酸、咖啡酸、阿魏酸和丹酚酸B的含量

目的本文旨在建立一种利用UPLC同时测定三叉神经散中绿原酸、咖啡酸、阿魏酸和丹酚酸B含量的方法。方法采用超高效液相色谱法,采用Kromat Universil XB C18色谱柱(150 mm×2.1 mm,3μm);流动相:乙腈-体积分数0.2%甲酸水溶液;流速:0.2 mL·min~(-1);检测波长:320 nm;进样量:1μL;柱温:30℃。结果绿原酸、咖啡酸、阿魏酸和丹酚酸B质量浓度分别在4.0~24.0 mg·L~(-1),0.2~1.2 mg·L~(-1),0.5~3.0 mg·L~(-1)和8.0~48.0 mg·L~(-1)内与峰面积呈良好的线性关系。绿原酸、咖啡酸、阿魏酸和丹酚酸B的平均回收率分别为(98.1±1.38)%、(97.7±1.10)%、(97.5±1.13)%和(98.3±1.39)%,RSD分别为1.41%、1.13%、1.16%和1.41%(n=9)。对三批样品进行含量测定,绿原酸、咖啡酸、阿魏酸和丹酚酸B的平均含量为(262.919±0.12)、(12.583±0.02)、(26.864±0.06)、(701.947±0.05)μg·g-1。结论本方法快速、简便、准确,可用于三叉神经散中绿原酸、咖啡酸、阿魏酸和丹酚酸B的含量测定。     

Studies on caffeic acid derivatives in medicinal plants

The components of Artemisia and Labiatae species were isolated and their chemical structures were determined. These compounds were found to have tannic activity for the first time. The activity of the compounds comes from the caffeic acid moiety in their molecules, not from structural features of usual tannins (hydrolyzable and condensed tannins). These also showed several other biological activities. Furthermore, the research on dimerization of caffeic acid derivatives was performed.     

咖啡酸苯乙酯纳米混悬剂的制备及其体外抑制乳腺癌细胞作用研究

目的 优化咖啡酸苯乙酯纳米混悬剂的处方,并考察其体外抑制乳腺癌细胞作用.方法 以泊洛沙姆188、蜂胶作为载体,采用反溶剂沉淀法制备,通过星点设计–效应面法优化最佳制备工艺参数,并考察咖啡酸苯乙酯纳米混悬剂的稳定性、载药量、包封率以及冻干保护剂的筛选,同时考察对4T1乳腺癌细胞的生长抑制作用和细胞摄取情况.结果 咖啡酸苯...