中国药理学与毒理学杂志
中國藥理學與毒理學雜誌
중국약이학여독이학잡지
CHINESE JOURNAL OF PHARMACOLOGY AND TOXICOLOGY
2014年
3期
408-414
,共7页
左靖%杨明%李淼%杜丽娜%金义光
左靖%楊明%李淼%杜麗娜%金義光
좌정%양명%리묘%두려나%금의광
吉西他滨%脂质衍生物%聚合物胶束%抗肿瘤药
吉西他濱%脂質衍生物%聚閤物膠束%抗腫瘤藥
길서타빈%지질연생물%취합물효속%항종류약
ge mcitabine%lipid derivatives%polymeric micelles%antineoplastic agents
目的：制备吉西他滨（Ge m）脂质衍生物及其聚合物胶束，克服Ge m体内易失活、不能口服和无靶向性的缺陷。方法合成N-苯甲酰-3′-乙酰吉西他滨（BAG）。用注入法制备载BAG的泊洛沙姆188胶束（BAG∶泊洛沙姆188＝10∶1，mol/mol）；用激光粒度和电位仪测定胶束的粒度和Zeta电位；胶束用负染法透射电镜法观察胶束微观形态。Gem或BAG聚合物胶束5，10，20，30，50，70和90μmol·L-1与人乳腺癌细胞MCF-7培养24，48和72 h，用MTT法测定对MCF-7细胞生长的抑制率。肝癌细胞H22移植瘤小鼠分别iv或ig给予Ge m 40 mg·kg -1或BAG聚合物胶束62 mg·kg -1，间隔2 d给药1次，共3次，末次给药后第2天处死小鼠，测定抑瘤率。结果经薄层色谱法、核磁共振氢谱和碳谱、红外光谱和质谱验证， BAG的结构正确。BAG 聚合物胶束外观为淡蓝色乳光的透明溶液，粒径为62．82 n m，Zeta 电位为-18．8 mV，电镜下呈现球状均匀分布。MTT法实验结果表明，Ge m和BAG 聚合物胶束分别与MCF-7细胞培养24，48和72 h，其抑制 MCF-7细胞生长的 IC50分别为40．6和90．0，5．0和14．9，5．0和13．6μmol·L-1。体内实验结果表明，Gem口服和注射组与模型对照组比较（P＜0．01）、BAG 聚合物胶束口服和注射组与泊洛尔姆188空白胶束组比较（P＜0．05，P＜0．01）均具有明显的抑瘤作用；作用强度， BAG聚合物胶束口服优于Ge m口服（P＜0．05），BAG聚合物胶束注射优于其口服（P＜0．05），BAG聚合物胶束注射与Gem注射作用相当。结论 Gem脂质衍生物能载入泊洛沙姆188聚合物胶束内。BAG聚合物胶束体外对MCF-7细胞生长有抑制作用，iv和ig 给药对小鼠H22移植瘤均具有抑瘤作用。BAG 聚合物胶束口服优于Ge m口服，提示BAG聚合物胶束有望开发成为新型的抗肿瘤口服制剂。
目的：製備吉西他濱（Ge m）脂質衍生物及其聚閤物膠束，剋服Ge m體內易失活、不能口服和無靶嚮性的缺陷。方法閤成N-苯甲酰-3′-乙酰吉西他濱（BAG）。用註入法製備載BAG的泊洛沙姆188膠束（BAG∶泊洛沙姆188＝10∶1，mol/mol）；用激光粒度和電位儀測定膠束的粒度和Zeta電位；膠束用負染法透射電鏡法觀察膠束微觀形態。Gem或BAG聚閤物膠束5，10，20，30，50，70和90μmol·L-1與人乳腺癌細胞MCF-7培養24，48和72 h，用MTT法測定對MCF-7細胞生長的抑製率。肝癌細胞H22移植瘤小鼠分彆iv或ig給予Ge m 40 mg·kg -1或BAG聚閤物膠束62 mg·kg -1，間隔2 d給藥1次，共3次，末次給藥後第2天處死小鼠，測定抑瘤率。結果經薄層色譜法、覈磁共振氫譜和碳譜、紅外光譜和質譜驗證， BAG的結構正確。BAG 聚閤物膠束外觀為淡藍色乳光的透明溶液，粒徑為62．82 n m，Zeta 電位為-18．8 mV，電鏡下呈現毬狀均勻分佈。MTT法實驗結果錶明，Ge m和BAG 聚閤物膠束分彆與MCF-7細胞培養24，48和72 h，其抑製 MCF-7細胞生長的 IC50分彆為40．6和90．0，5．0和14．9，5．0和13．6μmol·L-1。體內實驗結果錶明，Gem口服和註射組與模型對照組比較（P＜0．01）、BAG 聚閤物膠束口服和註射組與泊洛爾姆188空白膠束組比較（P＜0．05，P＜0．01）均具有明顯的抑瘤作用；作用彊度， BAG聚閤物膠束口服優于Ge m口服（P＜0．05），BAG聚閤物膠束註射優于其口服（P＜0．05），BAG聚閤物膠束註射與Gem註射作用相噹。結論 Gem脂質衍生物能載入泊洛沙姆188聚閤物膠束內。BAG聚閤物膠束體外對MCF-7細胞生長有抑製作用，iv和ig 給藥對小鼠H22移植瘤均具有抑瘤作用。BAG 聚閤物膠束口服優于Ge m口服，提示BAG聚閤物膠束有望開髮成為新型的抗腫瘤口服製劑。
목적：제비길서타빈（Ge m）지질연생물급기취합물효속，극복Ge m체내역실활、불능구복화무파향성적결함。방법합성N-분갑선-3′-을선길서타빈（BAG）。용주입법제비재BAG적박락사모188효속（BAG∶박락사모188＝10∶1，mol/mol）；용격광립도화전위의측정효속적립도화Zeta전위；효속용부염법투사전경법관찰효속미관형태。Gem혹BAG취합물효속5，10，20，30，50，70화90μmol·L-1여인유선암세포MCF-7배양24，48화72 h，용MTT법측정대MCF-7세포생장적억제솔。간암세포H22이식류소서분별iv혹ig급여Ge m 40 mg·kg -1혹BAG취합물효속62 mg·kg -1，간격2 d급약1차，공3차，말차급약후제2천처사소서，측정억류솔。결과경박층색보법、핵자공진경보화탄보、홍외광보화질보험증， BAG적결구정학。BAG 취합물효속외관위담람색유광적투명용액，립경위62．82 n m，Zeta 전위위-18．8 mV，전경하정현구상균균분포。MTT법실험결과표명，Ge m화BAG 취합물효속분별여MCF-7세포배양24，48화72 h，기억제 MCF-7세포생장적 IC50분별위40．6화90．0，5．0화14．9，5．0화13．6μmol·L-1。체내실험결과표명，Gem구복화주사조여모형대조조비교（P＜0．01）、BAG 취합물효속구복화주사조여박락이모188공백효속조비교（P＜0．05，P＜0．01）균구유명현적억류작용；작용강도， BAG취합물효속구복우우Ge m구복（P＜0．05），BAG취합물효속주사우우기구복（P＜0．05），BAG취합물효속주사여Gem주사작용상당。결론 Gem지질연생물능재입박락사모188취합물효속내。BAG취합물효속체외대MCF-7세포생장유억제작용，iv화ig 급약대소서H22이식류균구유억류작용。BAG 취합물효속구복우우Ge m구복，제시BAG취합물효속유망개발성위신형적항종류구복제제。
OBJECTIVE Topreparealipidderivativeofgemcitabine(Gem)anditspolymericmi-celles to overcome the disadvantages of Gem.METHODS N-benzyl-3′-acetyl-gemcitabine(BAG)was synthesized.A BAG-loaded poloxamer polymeric micelle (BAG∶poloxamer 188 =10∶1 ,mol/mol)was prepared using an injection method.The micelles were characterized with a laser particle size and elec-tric charge instru ment and negatively-stained trans mission electron microscopy.Hu man breast cancer cells MCF-7 were cultured with Gem or BAG polymeric micelles of 5,10,20,30,50,70,90 μmol·L-1 for 24,48 and 72 h,respectively.The inhibitory rate of cells was measured with an MTT method.The MCF-7 cytotoxicity of BAG polymeric micelles was investigated.A pharmacodynamic study was per-formed on the mice bearing mouse hepatocellular cancer cells H22.Intravenous (iv)and oral (ig)ad-ministration was used at the dose of Ge m 40 mg·kg -1 or BAG polymeric micelles 62 mg·kg -1 .The mice were administered on the 1 st,4th and 7th day and sacrificed on the 8th day.Tumor inhibitory rates were measured.RESULTS TheBAGstructurewasidentifiedbythinlayerchromatograph,1Hand13C NMR,infrared ray chromatograph and mass spectrum.The appearance of BAG micelles was a slightly blue suspension.The micelles were spheres according to the electron microscopic observation.Their size was 62.82 nm and the zeta potential was -18.8 mV.The half inhibition concentration (IC50)of Gem and BAG polymeric micelles was 40.6 and 90.0 μmol·L-1 ,5.0 and 14.9 μmol·L-1 ,5.0 and 1 3.6 μmol·L-1 at 24,48 and 72 h,respectively according to the MTT results.According to the in vivo results,compared with the tumor model group,Gem (ig),Gem (iv)and BAG polymeric micelles (iv and ig)had significant effect on the tumor weight of H22 cell xenograft mice (P<0.01 ).As for anti-tumor efficiency,BAG polymeric micelles (ig)were better than Gem (ig)(P<0.05);BAG polymeric micelles (iv)were better than BAG polymeric micelles (ig)(P<0.05),and BAG polymeric micelles (iv)were almostequaltoGem(iv).CONCLUSION ThelipidderivativeofGemcanbeloadedinthepoloxamer 1 88 polymeric micelles.BAG polymeric micelles show in vitro MCF-7 cell inhibition and in vivo inhibition of mouse H22 xerografts;iv or ig.BAG polymeric micelles (ig)show better anti-tumor effect than Gem (ig),indicating that BAG polymeric micelles are a promising novel anti-tumor oral preparation.
