The improved 3 step preparation of a key antitumor agent, 7 ethyl 10 hydroxycamptothecin(SN 38), which consists of ethylation, oxidation and photo chemical rearrangement, is described. The proposed reaction mech...The improved 3 step preparation of a key antitumor agent, 7 ethyl 10 hydroxycamptothecin(SN 38), which consists of ethylation, oxidation and photo chemical rearrangement, is described. The proposed reaction mechanism is also discussed.展开更多
A novel camptothecin analogue, 9-methyl-10-hydroxycamptothecin （4）, was unexpectedly synthesized from 10-hydroxycamptothecin in two steps. The key step included an efficient Mannich-type reaction. The overall yield ...A novel camptothecin analogue, 9-methyl-10-hydroxycamptothecin （4）, was unexpectedly synthesized from 10-hydroxycamptothecin in two steps. The key step included an efficient Mannich-type reaction. The overall yield was 47.2%. An ether analogue of 4, 9-methyl-10-benzylaminomethoxycamptothecin （5）, was also prepared. These new camptothecin analogues were evaluated for in vitro cytotoxicity against four human cancer cell lines, and exhibited more potent antitumor activities than contrals camptothecin and topotecan against several cancer cells.展开更多
The present method gives a detailed description for the development and validation of a simple stability indicating reverse phase liquid chromatographic method for 10-hydroxycamptothecin(10-HCTN) in the presence of it...The present method gives a detailed description for the development and validation of a simple stability indicating reverse phase liquid chromatographic method for 10-hydroxycamptothecin(10-HCTN) in the presence of its impurities namely Imp A and Imp B along with degradation products generated from forced degradation studies. The drug was subjected to stress conditions of hydrolysis (acid, base and neutral), oxidative, photolytic and thermal stress degradation. Degradation was observed when subjected to treatment with peroxides or under conditions normally used for typical acid and base hydrolysis. The drug was found to be stable under other stress conditions attempted such as photolytic and thermal. Successful separation and isolation of the drug from related impurities and degradation products formed under stress conditions was achieved on an Inertsil ODS-3V (250 mm × 4.6 mm, 5 μm) column using a phosphate buffer, acetonitrile, methanol and Nanopure water. The developed HPLC method was validated with respect to specificity, linearity, accuracy, precision, sensitivity, robustness and solution stability. The assay method was found to be linear in the range of 0.16 mg/mL to 0.24 mg/mL with a correlation coefficient of 0.999 and the linearity of the impurities was established from 0.02% (LOQ) to 0.3%. Recoveries of assay and impurities were found between 99.4% and 100.3%. The developed HPLC method can be used to determine the related substances and assay determinations of 10-HCTN and also to evaluate the quality and long term stability of production samples.展开更多
以喜树果为原料,对匀浆法提取喜树碱和10-羟基喜树碱的工艺进行了研究,确定了最佳的工艺条件:提取溶剂为体积分数55%的乙醇,匀浆时间为8 m in,料液比为1∶15(g/mL)。在此工艺条件下,喜树碱和10-羟基喜树碱的提取率分别为0.801‰和0.546...以喜树果为原料,对匀浆法提取喜树碱和10-羟基喜树碱的工艺进行了研究,确定了最佳的工艺条件:提取溶剂为体积分数55%的乙醇,匀浆时间为8 m in,料液比为1∶15(g/mL)。在此工艺条件下,喜树碱和10-羟基喜树碱的提取率分别为0.801‰和0.546‰。将该法与超声提取、回流提取、常温冷浸提取、水浴振荡提取进行了比较,结果表明,匀浆提取具有得率高、省时间等方面的优势,是一种高效提取喜树碱和10-羟基喜树碱的方法。展开更多
旨在研究10-羟基喜树碱(10-hydroxycamptothecin,HCPT)和白藜芦醇(Resveratrol,Res)对体外培养的小鼠恶性黑色素瘤B16F10细胞的增殖及黑色素合成抑制机理。利用MTT法、显微观察、L-Dopa氧化法、Na OH裂解法分析不同浓度HCPT和Res对细胞...旨在研究10-羟基喜树碱(10-hydroxycamptothecin,HCPT)和白藜芦醇(Resveratrol,Res)对体外培养的小鼠恶性黑色素瘤B16F10细胞的增殖及黑色素合成抑制机理。利用MTT法、显微观察、L-Dopa氧化法、Na OH裂解法分析不同浓度HCPT和Res对细胞增殖、细胞形态、酪氨酸酶活性及黑色素合成含量的影响。荧光半定量PCR方法(Semi-RT-PCR)分析该化合物对黑色素合成关键因子酪氨酸酶(TYR)和小眼相关转录因子(MITF)基因表达的影响。结果表明HCPT(40、80、120、160和200μmol/L)和Res(80、120、160和200μmol/L)能够通过诱导细胞凋亡抑制B16F10细胞的增殖,同时对酪氨酸酶活性和细胞黑色素生成具有明显抑制作用(P<0.05),且呈现浓度依赖性。另外,不同浓度的HCPT以及高浓度Res(120和160μmol/L)能够显著下调B16F10细胞TYR和MITF基因的mRNA水平。HCPT和Res可能通过诱导细胞凋亡抑制B16F10细胞的增殖,同时通过下调MITF基因转录,抑制TYR m RNA的表达及TYR酶活性,进而抑制细胞黑色素的生成。展开更多
目的从喜树组织中筛选产10-羟基喜树碱的内生真菌,并对其进行鉴定。方法从喜树(Camptotheca acuminateDecne)树皮中分离、纯化得到6株内生真菌,经摇瓶发酵后,采用TLC法、HPLC和MS分析菌丝的提取物;用插片法进行形态学观察,采用分子生物...目的从喜树组织中筛选产10-羟基喜树碱的内生真菌,并对其进行鉴定。方法从喜树(Camptotheca acuminateDecne)树皮中分离、纯化得到6株内生真菌,经摇瓶发酵后,采用TLC法、HPLC和MS分析菌丝的提取物;用插片法进行形态学观察,采用分子生物学方法对XK002菌株rDNA的ITS基因(ITS-5.8S rDNA)进行PCR扩增、测序;利用BLAST软件与GenBank数据库进行相似性分析,并用Clustal X1.81软件法构建系统发育树。结果内生真菌XK002可产10-羟基喜树碱;菌株XK002在PDA培养基上生长迅速,白色绒毛状,4d能长满直径为9cm培养皿,10d后开始形成黑色的子座,产生大量的分生孢子;ITS序列与Valsa mali Miyabe et Yamada相应序列的相似性为100%,基因库接受号(Accession No.)为FJ418170。结论根据形态学和分子生物学方法鉴定可知,XK002菌株为Valsa mali Miyabe et Yamada。展开更多
文摘The improved 3 step preparation of a key antitumor agent, 7 ethyl 10 hydroxycamptothecin(SN 38), which consists of ethylation, oxidation and photo chemical rearrangement, is described. The proposed reaction mechanism is also discussed.
基金the Natural Science Foundation of Jiangsu Province,China(No.BK2007168)
文摘A novel camptothecin analogue, 9-methyl-10-hydroxycamptothecin （4）, was unexpectedly synthesized from 10-hydroxycamptothecin in two steps. The key step included an efficient Mannich-type reaction. The overall yield was 47.2%. An ether analogue of 4, 9-methyl-10-benzylaminomethoxycamptothecin （5）, was also prepared. These new camptothecin analogues were evaluated for in vitro cytotoxicity against four human cancer cell lines, and exhibited more potent antitumor activities than contrals camptothecin and topotecan against several cancer cells.
文摘The present method gives a detailed description for the development and validation of a simple stability indicating reverse phase liquid chromatographic method for 10-hydroxycamptothecin(10-HCTN) in the presence of its impurities namely Imp A and Imp B along with degradation products generated from forced degradation studies. The drug was subjected to stress conditions of hydrolysis (acid, base and neutral), oxidative, photolytic and thermal stress degradation. Degradation was observed when subjected to treatment with peroxides or under conditions normally used for typical acid and base hydrolysis. The drug was found to be stable under other stress conditions attempted such as photolytic and thermal. Successful separation and isolation of the drug from related impurities and degradation products formed under stress conditions was achieved on an Inertsil ODS-3V (250 mm × 4.6 mm, 5 μm) column using a phosphate buffer, acetonitrile, methanol and Nanopure water. The developed HPLC method was validated with respect to specificity, linearity, accuracy, precision, sensitivity, robustness and solution stability. The assay method was found to be linear in the range of 0.16 mg/mL to 0.24 mg/mL with a correlation coefficient of 0.999 and the linearity of the impurities was established from 0.02% (LOQ) to 0.3%. Recoveries of assay and impurities were found between 99.4% and 100.3%. The developed HPLC method can be used to determine the related substances and assay determinations of 10-HCTN and also to evaluate the quality and long term stability of production samples.
文摘以喜树果为原料,对匀浆法提取喜树碱和10-羟基喜树碱的工艺进行了研究,确定了最佳的工艺条件:提取溶剂为体积分数55%的乙醇,匀浆时间为8 m in,料液比为1∶15(g/mL)。在此工艺条件下,喜树碱和10-羟基喜树碱的提取率分别为0.801‰和0.546‰。将该法与超声提取、回流提取、常温冷浸提取、水浴振荡提取进行了比较,结果表明,匀浆提取具有得率高、省时间等方面的优势,是一种高效提取喜树碱和10-羟基喜树碱的方法。
文摘旨在研究10-羟基喜树碱(10-hydroxycamptothecin,HCPT)和白藜芦醇(Resveratrol,Res)对体外培养的小鼠恶性黑色素瘤B16F10细胞的增殖及黑色素合成抑制机理。利用MTT法、显微观察、L-Dopa氧化法、Na OH裂解法分析不同浓度HCPT和Res对细胞增殖、细胞形态、酪氨酸酶活性及黑色素合成含量的影响。荧光半定量PCR方法(Semi-RT-PCR)分析该化合物对黑色素合成关键因子酪氨酸酶(TYR)和小眼相关转录因子(MITF)基因表达的影响。结果表明HCPT(40、80、120、160和200μmol/L)和Res(80、120、160和200μmol/L)能够通过诱导细胞凋亡抑制B16F10细胞的增殖,同时对酪氨酸酶活性和细胞黑色素生成具有明显抑制作用(P<0.05),且呈现浓度依赖性。另外,不同浓度的HCPT以及高浓度Res(120和160μmol/L)能够显著下调B16F10细胞TYR和MITF基因的mRNA水平。HCPT和Res可能通过诱导细胞凋亡抑制B16F10细胞的增殖,同时通过下调MITF基因转录,抑制TYR m RNA的表达及TYR酶活性,进而抑制细胞黑色素的生成。
文摘目的从喜树组织中筛选产10-羟基喜树碱的内生真菌,并对其进行鉴定。方法从喜树(Camptotheca acuminateDecne)树皮中分离、纯化得到6株内生真菌,经摇瓶发酵后,采用TLC法、HPLC和MS分析菌丝的提取物;用插片法进行形态学观察,采用分子生物学方法对XK002菌株rDNA的ITS基因(ITS-5.8S rDNA)进行PCR扩增、测序;利用BLAST软件与GenBank数据库进行相似性分析,并用Clustal X1.81软件法构建系统发育树。结果内生真菌XK002可产10-羟基喜树碱;菌株XK002在PDA培养基上生长迅速,白色绒毛状,4d能长满直径为9cm培养皿,10d后开始形成黑色的子座,产生大量的分生孢子;ITS序列与Valsa mali Miyabe et Yamada相应序列的相似性为100%,基因库接受号(Accession No.)为FJ418170。结论根据形态学和分子生物学方法鉴定可知,XK002菌株为Valsa mali Miyabe et Yamada。
