2-甲氧基雌二醇静脉乳剂的研究
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摘要
2-甲氧基雌二醇(2-methoxyestradiol,2-ME)是雌激素的天然代谢产物,具有抗肿瘤活性,目前,其抗肿瘤活性国外处于Ⅰ、Ⅱ期临床研究阶段,2-ME是一种浓度时间依赖性的抗肿瘤药物,在水中溶解度极低(约0.2206μg·ml~(-1)),口服吸收和剂量之间缺乏相关性,在体内又易快速羟基化、脱氢等生物变化产生非活性的代谢产物,导致半衰期较短,生物利用度较低。因而考虑将其制成乳剂以达到缓释及提高生物利用度的作用。本研究的目的是以2-ME为模型药物制备静脉注射乳剂,考察脂溶性药物制成O/W乳剂后的安全性,药效性及缓释靶向性,为2-ME的新剂型开发提供研究基础。
本文对2-ME静脉乳剂的处方和工艺进行了研究,测定了25℃时,2-ME在大豆油、水中的溶解度分别为1.05 mg·ml~(-1)、0.2206μg·ml~(-1),脂水分配系数logP为4.41。考察了乳剂处方中油相比例,乳化剂(蛋黄卵磷脂、泊洛沙姆)比例,药物与磷脂比例及乳化温度、乳化时间、均质压力及均质次数等影响乳化效果的处方及制备工艺因素,用正交设计及单因素法筛选出较优的处方及制备工艺。
对2-ME静脉注射乳剂药剂学性质进行了研究,测得2-ME乳剂的粒径为246.2±5.6nm;Zeta电位为-32.47±1.51 mv;pH值为7.32±0.12;黏度为1.423±0.005 mpa.S,符合2005版药典对乳剂的要求。通过离心稳定性,影响因素试验,冷冻-加热循环试验,加速试验及长期试验的研究,表明本研究所制备的乳剂稳定性较好,在冷藏条件下粒径无明显变化,有较好的贮存稳定性。通过不同的释药条件及释药方法对2-ME乳剂在体外的释放情况进行考察,2-ME对照液有突释现象,30 min达50%,2 h达到90%,以后缓慢释放,2-ME乳剂的累积释药量则在6 h才接近50%,30 h接近90%,可见在体外2-ME乳剂有一定的缓释效果。
通过对2-ME乳剂进行急性毒性、刺激性、过敏性以及溶血性检查,对其安全性进行初步评价。结果显示2-ME静脉注射乳剂毒性非常低,对兔耳静脉血管没有不良刺激作用,无溶血及过敏反应发生。采用MTT体外药敏实验,验证了2-ME乳剂对MCF-7细胞的增殖抑制作用,并且这种抑制作用随着时间及剂量的增加而增强。
以小鼠为模型动物,采用HPLC(荧光检测器)测定2-ME在血浆及各组织中的浓度,2-ME静脉注射乳剂在小鼠体内的药物代谢动力学规律;结果表明2-ME静脉注射乳剂具有一定的缓释及肺、肝、脾等组织靶向性。
2-Methoxyestradiol(2-ME),which is studied at clinical phaseⅠ,Ⅱabroad,is a natural estrogen metabolite with the characteristics anti-tumors activity.2-ME has the manners of time-concentration-dependent,low solubility(approximately 0.2206μg·ml~(-1));lack of correlation between oral absorption and dose;and it can rapidly produce inactive metabolites due to easy hydroxylation,dehydrogenation in vivo.The above causes resulted in shorter half-life and low oral bioavailability.The emulsion has the roles of delayed-release and increase bioavailability.The purpose of the study is to prepare o/w intravenous emulsion of 2-ME and investigate the safety,efficacy, delayed-release and targeted effect to tumors cells,and provide the research base for new formulations of 2-ME.
Formulation design and preparation technique of 2-ME intravenous emulsion was studied,the solubility of 2-ME in soybean oil and water were 1.05 mg·ml~(-1),0.2206μg·ml~(-1) at 25℃respestively,the lipid water partition coefficient(logP) was 4.41 at 25℃,so 2-ME could be dissolved in soybean oil to prepare emulsion.The effects of the percentage of soya bean oil,the percentage of emulsifier,the ratio of between phospholipids and poloxamer,the ratio of between drug and phospholipid and the influennces of emulsification temperatures,emulsification time,homogeneous stress and homogeneous times on the emulsion quality were studied,Then based on the results of single factor analysis,the formulation and the preparation techniques were optimized by orthogonal design.
The pharmaceutical properties of 2-ME intravenous emulsion were as followings: particle size 246.2±5.6 nm,Zeta potential -32.47±1.51 mv,pH value 7.32±0.12, viscosity 1.423±0.005 mpa.S.The physicochemical properties of 2-ME intravenous emulsion were coincident with the Chinese Pharmacopoeia.The centrifugal test, impact factor test,frozen-heating cycle test,accelerated test and long-term test were conducted to investigate the stability of emulsion,the results showed that the particle size didn't increase significantly,indicating that the emulsion had good stability in frozen conditions.The accumulated release amount of 2-ME control solution was up to 50%in 30 min,up to 90%in 2 h,the accumulated release of 2-ME emulsion was about 50%in 6 h,about 90%in 30 h.Showing that 2-ME emulsion had slow-release effect compared to 2-ME control solution.
The preliminary safety of 2-ME emulsion was evaluate by acute toxicity test, vascular irritation test,haematolysis and hypersensitiveness test.The results showed that the emulsion had no obvious toxicity,irritation,haemocytolysis and hypersensitiveness.The effects of the 2-ME control solution and 2-ME emulsion on the MCF-7 cell proliferation inhibition were investigated by use of MTT method, indicating that inhibition effect was increased with time and dose increment, 2-ME-emulsion had slow-release effect compared to 2 -ME control solution.
2-ME blood concentration of mice were determined by HPLC with fluorescence detector to explore pharmacokinetics parameters for 2-ME intravenous emulsion and 2-ME control;At the same time,the 2-ME distribution in mice were investigated for 2-ME intravenous emulsion.Showing that 2-ME emulsion has a sustained-release and tissue targeting in lung,liver,spleen compared to 2-ME control solution.
本文对2-ME静脉乳剂的处方和工艺进行了研究,测定了25℃时,2-ME在大豆油、水中的溶解度分别为1.05 mg·ml~(-1)、0.2206μg·ml~(-1),脂水分配系数logP为4.41。考察了乳剂处方中油相比例,乳化剂(蛋黄卵磷脂、泊洛沙姆)比例,药物与磷脂比例及乳化温度、乳化时间、均质压力及均质次数等影响乳化效果的处方及制备工艺因素,用正交设计及单因素法筛选出较优的处方及制备工艺。
对2-ME静脉注射乳剂药剂学性质进行了研究,测得2-ME乳剂的粒径为246.2±5.6nm;Zeta电位为-32.47±1.51 mv;pH值为7.32±0.12;黏度为1.423±0.005 mpa.S,符合2005版药典对乳剂的要求。通过离心稳定性,影响因素试验,冷冻-加热循环试验,加速试验及长期试验的研究,表明本研究所制备的乳剂稳定性较好,在冷藏条件下粒径无明显变化,有较好的贮存稳定性。通过不同的释药条件及释药方法对2-ME乳剂在体外的释放情况进行考察,2-ME对照液有突释现象,30 min达50%,2 h达到90%,以后缓慢释放,2-ME乳剂的累积释药量则在6 h才接近50%,30 h接近90%,可见在体外2-ME乳剂有一定的缓释效果。
通过对2-ME乳剂进行急性毒性、刺激性、过敏性以及溶血性检查,对其安全性进行初步评价。结果显示2-ME静脉注射乳剂毒性非常低,对兔耳静脉血管没有不良刺激作用,无溶血及过敏反应发生。采用MTT体外药敏实验,验证了2-ME乳剂对MCF-7细胞的增殖抑制作用,并且这种抑制作用随着时间及剂量的增加而增强。
以小鼠为模型动物,采用HPLC(荧光检测器)测定2-ME在血浆及各组织中的浓度,2-ME静脉注射乳剂在小鼠体内的药物代谢动力学规律;结果表明2-ME静脉注射乳剂具有一定的缓释及肺、肝、脾等组织靶向性。
2-Methoxyestradiol(2-ME),which is studied at clinical phaseⅠ,Ⅱabroad,is a natural estrogen metabolite with the characteristics anti-tumors activity.2-ME has the manners of time-concentration-dependent,low solubility(approximately 0.2206μg·ml~(-1));lack of correlation between oral absorption and dose;and it can rapidly produce inactive metabolites due to easy hydroxylation,dehydrogenation in vivo.The above causes resulted in shorter half-life and low oral bioavailability.The emulsion has the roles of delayed-release and increase bioavailability.The purpose of the study is to prepare o/w intravenous emulsion of 2-ME and investigate the safety,efficacy, delayed-release and targeted effect to tumors cells,and provide the research base for new formulations of 2-ME.
Formulation design and preparation technique of 2-ME intravenous emulsion was studied,the solubility of 2-ME in soybean oil and water were 1.05 mg·ml~(-1),0.2206μg·ml~(-1) at 25℃respestively,the lipid water partition coefficient(logP) was 4.41 at 25℃,so 2-ME could be dissolved in soybean oil to prepare emulsion.The effects of the percentage of soya bean oil,the percentage of emulsifier,the ratio of between phospholipids and poloxamer,the ratio of between drug and phospholipid and the influennces of emulsification temperatures,emulsification time,homogeneous stress and homogeneous times on the emulsion quality were studied,Then based on the results of single factor analysis,the formulation and the preparation techniques were optimized by orthogonal design.
The pharmaceutical properties of 2-ME intravenous emulsion were as followings: particle size 246.2±5.6 nm,Zeta potential -32.47±1.51 mv,pH value 7.32±0.12, viscosity 1.423±0.005 mpa.S.The physicochemical properties of 2-ME intravenous emulsion were coincident with the Chinese Pharmacopoeia.The centrifugal test, impact factor test,frozen-heating cycle test,accelerated test and long-term test were conducted to investigate the stability of emulsion,the results showed that the particle size didn't increase significantly,indicating that the emulsion had good stability in frozen conditions.The accumulated release amount of 2-ME control solution was up to 50%in 30 min,up to 90%in 2 h,the accumulated release of 2-ME emulsion was about 50%in 6 h,about 90%in 30 h.Showing that 2-ME emulsion had slow-release effect compared to 2-ME control solution.
The preliminary safety of 2-ME emulsion was evaluate by acute toxicity test, vascular irritation test,haematolysis and hypersensitiveness test.The results showed that the emulsion had no obvious toxicity,irritation,haemocytolysis and hypersensitiveness.The effects of the 2-ME control solution and 2-ME emulsion on the MCF-7 cell proliferation inhibition were investigated by use of MTT method, indicating that inhibition effect was increased with time and dose increment, 2-ME-emulsion had slow-release effect compared to 2 -ME control solution.
2-ME blood concentration of mice were determined by HPLC with fluorescence detector to explore pharmacokinetics parameters for 2-ME intravenous emulsion and 2-ME control;At the same time,the 2-ME distribution in mice were investigated for 2-ME intravenous emulsion.Showing that 2-ME emulsion has a sustained-release and tissue targeting in lung,liver,spleen compared to 2-ME control solution.
引文
[1]Leonard R.Axelrod,P.Narasirnha Rao,Joseph W.Goldzieher.The conversion of 2-hydroxyestradiol-17β to 2-hydroxy and 2-methoxy metabolites in human urine.Archives of Biochemistry and Biophysics,1961,94(2):265-268.
[2]Christopher Sweeney,Glenn Liu,Constantin Yiannoutsos,et al.A Phase ⅡMulticenter,Randomized,Double-Blind,Safety Trial Assessing the Pharmacokinetics,Pharmacodynamics,and Efficacy of Oral 2-Methoxyestradiol Capsules in Hormone-Refractory Prostate Cancer.Clinical Cancer Res 2005,18(11):6625-6633.
[3]黄豪博.2-甲氧基雌二醇的抗肿瘤作用及机制研究.国外医学输血及血液分册,2006,29(3):204-206.
[4]Giuseppina Basinia,Simona Bussolatia,Sujen Eleonora Santinia,Federica Bianchib,Antiangiogenesis in swine ovarian follicle:A potential role for 2-methoxyestradiol.steroids 2007,72(2):660-665.
[5]Tara E.Sutherland1,Robin L.Anderson2,Richard A.Hughesl.2-Methoxyestradiol-a unique blend of activities generating a new class of anti-tumour/anti-inflammatory agents.Drag Discovery Today,2007,12(4):13-14.
[6]Ying Dai,Meifeng Xu,Yigang Wang et al.HIF-1α induced-VEGF overexpression in bone marrow stem cells protects cardio-myo-cytes against ischemiav.Journal of Molecular and Cellular Cardiology,2007,42(5):1036-1044.
[7]Gregory E.Agoston,Jamshed H.Shah,Theresa M.et al.Synthesis and structure activity relationships of 16-modified analogs of 2-methoxyestradiol.Bioorganic &Medicinal Chemistry,2007,15(2):7524-7537.
[8]Junhao Yan,Chunhua Chen,Jilian Lei et al.2-methoxyestradiol reduces cerebral vasospasm after 48 hours of experimental subarachnoid hemorrhage in rats.Experimental Neurology,2006,202(6):348-356.
[9]Hei-Cheul Jeung,Xiao-Fang Che,Misako Haraguchi.Thymidine phosphorylase suppresses apoptosis induced by microtubule-interfering agents.Biochemical Pharmacology,2005,70(5):13-21.
[10]Klauber N,Parangi S,Flynn E,et al.Inhibition of angiogenesis and breast cancer in mice by the microtubule inhibitors 2-methoxyestradiol and taxol.Cancer Res,1997,57(3):81-86.
[11]Sweeney CJ,Miller KD,Sissons SE,et al.The antiangiogenic property of docetaxel is synergistic with a recombinant humanized monoclonal antibody against vascular endothelial growth factor or 2-methoxyestradiol but antagonized by endothelial growth factors.Cancer Res,2001,61(4):3369-3372.
[12]Yue TL,Wang X,Louden CS,et al.2-Methoxyestradiol,an endogenous estrogen metabolite,induces apoptosis in endothelial cells and inhibits angiogenesis:possible role for stress-activated protein kinase signaling pathway and Fas expression.Mol Pharmacol,1997,51(3):951-962.
[13]Zhou NN,Zhu XF,Zhou JM,et al.2-Methoxyestradiol induces cell cycle arrest and apoptosis of nasopharyngeal carcinoma cells.Acta Pharmacol Sin,2004,25(11):1515-1520.
[14]Ricker JL,Chen Z,Yang XP,et al.2-methoxyestradiol inhibits hypoxiainducible factor 1alpha,tumor growth,and angiogenesis and augments paclitaxel efficacy in head and neck squamous cell carcinoma.Clin Cancer Res,2004,24(10):8665-8673.
[15]Sandhya K.Nair,Arti Verma,T.J.Thomas,et al.Synergistic apoptosis of MCF-7breast cancer cells by 2-methoxyestradiol and bis(ethyl)norspermine.Cancer Letters 2007,250(5):312-322.
[16]Jian Hou,Hong Xiong,Weiran Gao,et al.2-Methoxyestradiol at low dose induces differentiation of myeloma cells.Leukemia Research,2005,29(6):1059-1067.
[17]LaVallee TM,Zhan XH,Johnson MS,et al.2-methoxyestradiol up-regulates death receptor and induces apoptosis through activation of the extrinsic pathway.Cancer Res,2003,63(2):468-475.
[18]Banerjee SN,Sengupta K,Banerjee S,et al.2-Methoxyestradiol exhibits a biphasic effect on VEGF2A in tumor cells and upregulation is mediated through ER alpha:a possible signaling pathway associated with the impact of 2-ME on proliferative cells.Neoplasia,2003,5(5):417-426.
[19]Chauhan D,Li G,Hideshima T,et al.JNK dependent release of mitochondrial protein,Smac,during apoptosis in multiple myeloma(MM)cells.J Biol Chem,2003, 278 (20) :17593-17596.
[20] Davoodpour P, Landstrom M. 2-Methoxyestradiol induced apoptosis in prostate cancer cells requires Smad.J Biol Chem, 2005,280(15) :14773-14779.
[21] Gao N, Rahmani M, Dent P, et al. 2-Methoxyestradiol induced apoptosis in human leukemia cells proceeds through a reactive oxygen species and Akt-dependent process. Oncogene, 2005, 24 (23): 3797-3809.
[22] Amorino GP, Freeman ML, Choy H. Enhancement of radiation effects in vitro by the estrogen metabolite 2-methoxyestradiol. Radiat Res, 2000,153(4) :384-391.
[23] Cushman M, Mohanakrishnan AK, Hollingshead M, et al. The effect of exchanging various substituents at the 2-position of 2-metheoxyestradiol on cytotocity in human cancer cell cultures and inhibition of tubulin polymerization [J]. Med Chem,2002, 45(21): 4748-4754.
[24] Jehana James, Daryl J. Murry, Anthony M. Treston et al. Phase I safety,pharmacokinetic and pharmacodynamic studies of 2-methoxyestradiol alone or in combination with docetaxel in patients with locally recurrent or metastatic breast cancer. Invest New Drugs , 2006, 25(3): 41-48.
[25] D. Chan, T LaVaiiee, Z Zhang, et al. 2-methoxyestradiol and its new derivatives inhibited lung cancer cells in vitro and prolonged survivals in atttymic nude rats at orthotopic lung tumors in vivo. Poster Discussions Basic Science Cell, 2005: 868.
[26] Dingli D, Timm M, Russell SJ, et al. Promising preclinical activity of 2-metheoxyestradiol in multiple myeloma. [J] Clin Cancer Res, 2002, 12(8):3948-3954.
[27] Schumacher G, Kataoka M, Roth et al. Potent anti-tumor activity of 2-methoxyestradiol in human pancreatic cancer cell lines. Clin Cancer Res, 1999,12(5): 493-499.
[28] Jyh-Ming Chowa, Chien-Ru Liub, Che-Pin Line, et al. Downregulation of c-Myc determines sensitivity to 2-methoxyestradiol-induced apoptosis in human acute myeloid leukemia .Experimental Hematology, 2008, 36(4): 140-148.
[29] Stevan P, Xichen Zhang, Edwin K, et al. 2-Methoxyestradiol mediates the protective effects of estradiol in monocrotaline-induced pulmonary hypertension Vascular. Pharmacology, 2006,45(5) :358—367.
[30] Agnieszka Lisa, Michael J. Ciesielskia, Tara A. Baronea,et al.2-Methoxyestradiol inhibits proliferation of normal and neoplastic glial cells, and induces cell death, in vitro.Cancer Letters, 2004,213(34) :57-65.
[31] Huober JB, Nakamura S, Meyn R, et al. Oral administration of an estrogen metabolite induced potentiation of radiation antitumor effects in presence of wild type p53 in non-small-cell lung cancer[J]. Int J Radiat Oncol Biol Phys, 2000, 48(4):1127-1137.
[32] Sweeney C, Liu G, Yiannoutsos C, et al. A phase II multicenter, randomized,double blind, safety trial assessing the pharmacokinetics, pharmacodynamics, and efficacy of oral 2-methoxyestradiol capsules in hormone refractory prostate cancer.Clin Cancer Res, 2005,18(11): 6625-6633.
[33] Goud H. Desai, Susan R. Mallery, Steven P. Schwendeman. Effect of formulation parameters on 2-methoxyestradiol release from injectable cylindrical poly(DL-lactide-co-glycolide) implants. European Journal of Pharmaceutics and Biopharmaceutics, 2008, 70(12) :187-198.
[34] MacFie J. The development of fat emulsions [J]. Nut rition ,1999 , 728 (15) :641.
[35] A.Akkar , R.H.Miiller. Formulation of intravenous Carbamazepine emulsions by SolEmuls technology. European Journal of Pharmaceutics and Biopharmaceutics, 2003,234(55): 305-312.
[36] Regina G. Kelmann, Gislaine Kuminek, Helder F. Carbamazepine parenteral nanoemulsions prepared by spontaneous emulsification process. International Journal of Pharmaceutics, 2007, 342(2): 231-239.
[37] Jorge Medina, Angeles Salvado , Alfonso del Pozo. Use of ultrasound to prepare lipid emulsions of lorazepam for intravenous injection. International Journal of Pharmaceutics, 2001, 216(12): 1-8.
[38] Wang H, Yang G, Liang WQ. Preparation of the dry emulsion of bupivicaine hydrochloride [J]. Strait Pharmaceutical Journal, 2004, 16(4): 21-22.
[39] Lu YT, Dong Y, Gao CZ.. Clinical efficiency of bruceolic oil emulsion on curing liver cancer of advanced stage [J]. Modern Preventive Medicine, 2005, 32(7): 845-846.
[40]吴玉波,张永恒,孙世中等.喜树碱精氨酸静脉乳剂药物动力学及组织分布的研究[J].中国医院药学杂志,1994,14(9):406-408.
[41]高晓黎,毕殿洲,孙殿甲,等,去氢骆驼蓬碱静脉注射乳剂在动物体内的药代动力学[J].药学学报,2000,35(3):224-227
[42]高晓黎,程利勇,孙殿甲,等.定量评价去氢骆驼蓬碱注射用乳剂的组织靶向[J].药学学报,2000,35(2):142-145.
[43]陈瑞珠,汪国玮,马晋隆,等.蒿甲醚靶向性静脉乳剂的研究[J].上海医药,1997,23(1):28.
[44]Tian X,Zhu JB,Wang JS.Preparation of tetrandrine lipid emulsion and its therapeutic effect on bleomycin-induced pulmonary fibro sis in Rats[J].China Pharma Univ,2005,36(3):225-229.
[45]Jiang WS.Preparation of allicin lipid emulsion[P].CN:1579375,2005-02-16.
[46]Guo T,Chu XQ,Song HT,et al.Preparation of garlic oil submicron emulsion for intravenous administration[J].Chinese Journal of Pharmaceuticals,2005,40(8):602-605.
[47]Constantinides PP,Lambert KJ,Tustian AK,et al.Formulation development and antitumor activity of a filter-sterilizable emulsion of Paclitaxe[J].Pharm Res,2000,17(2):175-182.
[48]Adwoa O.Nornoo,David W.Osborne,Diana S.-L.Chow.Cremophor-free intravenous microemulsions for paclitaxel:Formulation,cytotoxicity and hemolysis.International Journal of Pharmaceutics,2008,349(2):108-116.
[49]Raul C.Maranh(?)o,Elaine R.Tavares,Amanda F.Padoveze.Paclitaxel associated with cholesterol-rich nanoemulsions promotes atherosclerosis regression in the rabbit.Atherosclerosis,2008,197(2):959-966.
[50]Akkar A,Muller RH.Intravenous itraconazole emulsions produced by SolEmuls technology[J].Eur J Pharm Biopharm,2003,56(1):29-36.
[51]Brime B,Moreno MA,Frutos G,et al.Amphotericin B in oil-water lecithin-based microemulsions:formulation ang toxicity evaluation[J].J Pharm Sci,2002,91(4):1178-1185.
[52]Wang Y,Cory AL.A novel stable supersaturated submicron lipid emulsion of tirilazad[J].Pharm Dev Technol,1999,4(3):333-345.
[53]Jumaa M,Muller BW.Development of a novel parenteral formulation for tetrazepam using a lipid emulsion[J].Drug Dev Ind Pharm,2001,27(10):1115-1121.
[54]Yan Lu,YanJiao Wang,Xing Tang.Formulation and thermal sterile stability of a less painful intravenous clarithromycin emulsion containing vitamin E.International Journal of Pharmaceutics,2008,346(2):47-56.
[55]Yan Lu,Yu Zhang,Ziyi Yang,et al.Formulation of an intravenous emulsion loaded with a clarithromycin-phospholipid complex and its pharmacokinetics in rats.International Journal of Pharmaceutics,2009,366(2):160-169.
[56]刘琳琳.氟尿嘧啶乳剂研制与临床应用.临床药学,2005,23(2):4
[57]T.P.Formariz,V.H.V.Sarmento,A.A.Silva-Junior,et al.Doxorubicin biocompatible O/W microemulsion stabilized by mixed surfactant containing soya phosphatidylcholine.Colloids and Surfaces B:Biointerfaces,2006,51(1):54-57.
[58]Shaimaa S.Ibrahim,Gehanne A.S.Awad,Ahmed Geneidi,et al.Comparative effects of different cosurfactants on sterile prednisolone acetate ocular submicron emulsions stability and release.Colloids and Surfaces B:Biointerfaces,2009,69(2):225-231.
[59]Panayiotis P,Alex Tustian,Dean R.Tocol emulsions for drug solubilization and parenteral delivery.Advanced Drug Delivery Reviews,2004,254(56):1243-1255.
[60]Feng Hung,Lang Fang,Mei-Hui Liao,et al.The effect of oil components on the physicochemical properties and drug delivery of emulsions:Tocol emulsion versus lipid emulsion.International Journal of Pharmaceutics,2007,335(2):193-202.
[61]Jie Li,Shufang Nie,Xingang Yang,et al.Optimization of tocol emulsions for the intravenous delivery of clarithromycin.International Journal of Pharmaceutics,2008,356(2):282-299.
[62]Shuai Shi,Hao Chen,Yue Cui,et al.Formulation,stability and degradation kinetics of intravenous cinnarizine lipid emulsion..International Journal of Pharmaceutics,2009,345(2):240-245.
[63]Archie J.Sirianni,Kevin C.Osterhoudt,Diane P.Calello,et al.Use of Lipid Emulsion in the Resuscitation of a Patient With Prolonged Cardiovascular Collapse After Overdose of Bupropion and Lamotrigine. Annals of Emergency Medicine, 2008,51(4): 412-415.
[64] Srinivas Ganta, James W. Paxton, Bruce C. Baguley, et al. Pharmacokinetics and pharmacodynamics of chlorambucil delivered in parenteral emulsion. International Journal of Pharmaceutics, 2008, 360(2): 115-121.
[65]Mukesh Kumar, Ambikanandan Misra, A.K. Babbar, et al. Intranasal nanoemulsion based brain targeting drug delivery system of risperidone. International Journal of Pharmaceutics, 2008, 358(2): 285-297.
[66] Joung Wang, K.C. Sung, Hui Yeh, Jia-You Fang. The delivery and antinociceptive effects of morphine and its ester prodrugs from lipid emulsions. International Journal of Pharmaceutics, 2008, 353(2): 95-104.
[67] Joung Wang, K.C. Sung, Oliver Yoa-Pu Hu, Hui Yeh. .Submicron lipid emulsion as a drug delivery system for nalbuphine and its prodrugs. Journal of Controlled Release, 2006,115(2): 140-149.
[68] Granato D, Blum S, Rossle C, et al, Effects of parenteral lipid emulsions with different fatty acid composition on immune cell functions in vitro[J]. JPEN J Parenter Nutr, 2002,24(2): 113-118.
[69] Nordenstrom J, Thorne A, Olivercrona A. Metabolic effects of infusion of a structured-triglyceride emulsion in healthy subjects [J]. Nurtrition,1995,11(3):269-274.
[70] Kawaguchi, Shimokawa, Fumiyoshi. Physicochemical properties of structured phosphatidylcholine in drug carrier lipid emulsions for drug delivery systems. Colloids and Surfaces B: Biointerfaces, 2008, 62(1): 130-135.
[71] M. Wulff-P(?)rez, A. Torcello-Gomez, M.J. G(?)lvez-Ruiz.. Stability of emulsions for parenteral feeding: Preparation and characterization of o/w nanoemulsions with natural oils and Pluronic f68 as surfactant. Food Hydrocolloids, 2009, 23 (4): 1096-1102.
[72] Michele Trotta, Franco Pattarino, Terenzio Ignoni. Stability of durg-carrier emulsions containing phosphatidylcholine mixtures. European Journal of Pharmaceutics and Biopharmaceutics, 2002, 53(5): 203-208.
[73] I. Cuellar, J. Bullon, A.M.Forgarini. More efficient preparation of parenteral emulsions or how to improve a pharmaceutical recipe by formulation.Chemical Engineering Science,2005,60(7):2127-2134.
[74]Seid Mahdi Jafari,Yinghe He,Bhesh Bhandari.Production of sub-micron emulsions by ultrasound and microfluidization techniques.Journal of Food Engineering,2007,82(4):478-488.
[75]R.H.Muller,S.Schmidt,I.Buttle,et al.SolEmuls-novel technology for the formulation of i.v.Emulsions with poorly soluble drugs.International Journal of Pharmaceutics,2004,269(2):293-302.
[76]陆彬.药物新剂型与新技术.北京:人民卫生出版社,2005年版.
[77]国家药典委员会.中华人民共和国药典.化学工业出版社,2005年版二部.
[78]《化学药物急性毒性研究技术指导原则》课题研究组.化学药物急性毒性研究技术指导原则.2004年版.
[79]《化学药物刺激性、过敏性和溶血性研究技术指导原则》课题研究组.化学药物刺激性、过敏性和溶血性研究技术指导原则.2004年版.
[80]司徒振强.《细胞培养技术》.世界图书出版西安公司,2007年版.
[2]Christopher Sweeney,Glenn Liu,Constantin Yiannoutsos,et al.A Phase ⅡMulticenter,Randomized,Double-Blind,Safety Trial Assessing the Pharmacokinetics,Pharmacodynamics,and Efficacy of Oral 2-Methoxyestradiol Capsules in Hormone-Refractory Prostate Cancer.Clinical Cancer Res 2005,18(11):6625-6633.
[3]黄豪博.2-甲氧基雌二醇的抗肿瘤作用及机制研究.国外医学输血及血液分册,2006,29(3):204-206.
[4]Giuseppina Basinia,Simona Bussolatia,Sujen Eleonora Santinia,Federica Bianchib,Antiangiogenesis in swine ovarian follicle:A potential role for 2-methoxyestradiol.steroids 2007,72(2):660-665.
[5]Tara E.Sutherland1,Robin L.Anderson2,Richard A.Hughesl.2-Methoxyestradiol-a unique blend of activities generating a new class of anti-tumour/anti-inflammatory agents.Drag Discovery Today,2007,12(4):13-14.
[6]Ying Dai,Meifeng Xu,Yigang Wang et al.HIF-1α induced-VEGF overexpression in bone marrow stem cells protects cardio-myo-cytes against ischemiav.Journal of Molecular and Cellular Cardiology,2007,42(5):1036-1044.
[7]Gregory E.Agoston,Jamshed H.Shah,Theresa M.et al.Synthesis and structure activity relationships of 16-modified analogs of 2-methoxyestradiol.Bioorganic &Medicinal Chemistry,2007,15(2):7524-7537.
[8]Junhao Yan,Chunhua Chen,Jilian Lei et al.2-methoxyestradiol reduces cerebral vasospasm after 48 hours of experimental subarachnoid hemorrhage in rats.Experimental Neurology,2006,202(6):348-356.
[9]Hei-Cheul Jeung,Xiao-Fang Che,Misako Haraguchi.Thymidine phosphorylase suppresses apoptosis induced by microtubule-interfering agents.Biochemical Pharmacology,2005,70(5):13-21.
[10]Klauber N,Parangi S,Flynn E,et al.Inhibition of angiogenesis and breast cancer in mice by the microtubule inhibitors 2-methoxyestradiol and taxol.Cancer Res,1997,57(3):81-86.
[11]Sweeney CJ,Miller KD,Sissons SE,et al.The antiangiogenic property of docetaxel is synergistic with a recombinant humanized monoclonal antibody against vascular endothelial growth factor or 2-methoxyestradiol but antagonized by endothelial growth factors.Cancer Res,2001,61(4):3369-3372.
[12]Yue TL,Wang X,Louden CS,et al.2-Methoxyestradiol,an endogenous estrogen metabolite,induces apoptosis in endothelial cells and inhibits angiogenesis:possible role for stress-activated protein kinase signaling pathway and Fas expression.Mol Pharmacol,1997,51(3):951-962.
[13]Zhou NN,Zhu XF,Zhou JM,et al.2-Methoxyestradiol induces cell cycle arrest and apoptosis of nasopharyngeal carcinoma cells.Acta Pharmacol Sin,2004,25(11):1515-1520.
[14]Ricker JL,Chen Z,Yang XP,et al.2-methoxyestradiol inhibits hypoxiainducible factor 1alpha,tumor growth,and angiogenesis and augments paclitaxel efficacy in head and neck squamous cell carcinoma.Clin Cancer Res,2004,24(10):8665-8673.
[15]Sandhya K.Nair,Arti Verma,T.J.Thomas,et al.Synergistic apoptosis of MCF-7breast cancer cells by 2-methoxyestradiol and bis(ethyl)norspermine.Cancer Letters 2007,250(5):312-322.
[16]Jian Hou,Hong Xiong,Weiran Gao,et al.2-Methoxyestradiol at low dose induces differentiation of myeloma cells.Leukemia Research,2005,29(6):1059-1067.
[17]LaVallee TM,Zhan XH,Johnson MS,et al.2-methoxyestradiol up-regulates death receptor and induces apoptosis through activation of the extrinsic pathway.Cancer Res,2003,63(2):468-475.
[18]Banerjee SN,Sengupta K,Banerjee S,et al.2-Methoxyestradiol exhibits a biphasic effect on VEGF2A in tumor cells and upregulation is mediated through ER alpha:a possible signaling pathway associated with the impact of 2-ME on proliferative cells.Neoplasia,2003,5(5):417-426.
[19]Chauhan D,Li G,Hideshima T,et al.JNK dependent release of mitochondrial protein,Smac,during apoptosis in multiple myeloma(MM)cells.J Biol Chem,2003, 278 (20) :17593-17596.
[20] Davoodpour P, Landstrom M. 2-Methoxyestradiol induced apoptosis in prostate cancer cells requires Smad.J Biol Chem, 2005,280(15) :14773-14779.
[21] Gao N, Rahmani M, Dent P, et al. 2-Methoxyestradiol induced apoptosis in human leukemia cells proceeds through a reactive oxygen species and Akt-dependent process. Oncogene, 2005, 24 (23): 3797-3809.
[22] Amorino GP, Freeman ML, Choy H. Enhancement of radiation effects in vitro by the estrogen metabolite 2-methoxyestradiol. Radiat Res, 2000,153(4) :384-391.
[23] Cushman M, Mohanakrishnan AK, Hollingshead M, et al. The effect of exchanging various substituents at the 2-position of 2-metheoxyestradiol on cytotocity in human cancer cell cultures and inhibition of tubulin polymerization [J]. Med Chem,2002, 45(21): 4748-4754.
[24] Jehana James, Daryl J. Murry, Anthony M. Treston et al. Phase I safety,pharmacokinetic and pharmacodynamic studies of 2-methoxyestradiol alone or in combination with docetaxel in patients with locally recurrent or metastatic breast cancer. Invest New Drugs , 2006, 25(3): 41-48.
[25] D. Chan, T LaVaiiee, Z Zhang, et al. 2-methoxyestradiol and its new derivatives inhibited lung cancer cells in vitro and prolonged survivals in atttymic nude rats at orthotopic lung tumors in vivo. Poster Discussions Basic Science Cell, 2005: 868.
[26] Dingli D, Timm M, Russell SJ, et al. Promising preclinical activity of 2-metheoxyestradiol in multiple myeloma. [J] Clin Cancer Res, 2002, 12(8):3948-3954.
[27] Schumacher G, Kataoka M, Roth et al. Potent anti-tumor activity of 2-methoxyestradiol in human pancreatic cancer cell lines. Clin Cancer Res, 1999,12(5): 493-499.
[28] Jyh-Ming Chowa, Chien-Ru Liub, Che-Pin Line, et al. Downregulation of c-Myc determines sensitivity to 2-methoxyestradiol-induced apoptosis in human acute myeloid leukemia .Experimental Hematology, 2008, 36(4): 140-148.
[29] Stevan P, Xichen Zhang, Edwin K, et al. 2-Methoxyestradiol mediates the protective effects of estradiol in monocrotaline-induced pulmonary hypertension Vascular. Pharmacology, 2006,45(5) :358—367.
[30] Agnieszka Lisa, Michael J. Ciesielskia, Tara A. Baronea,et al.2-Methoxyestradiol inhibits proliferation of normal and neoplastic glial cells, and induces cell death, in vitro.Cancer Letters, 2004,213(34) :57-65.
[31] Huober JB, Nakamura S, Meyn R, et al. Oral administration of an estrogen metabolite induced potentiation of radiation antitumor effects in presence of wild type p53 in non-small-cell lung cancer[J]. Int J Radiat Oncol Biol Phys, 2000, 48(4):1127-1137.
[32] Sweeney C, Liu G, Yiannoutsos C, et al. A phase II multicenter, randomized,double blind, safety trial assessing the pharmacokinetics, pharmacodynamics, and efficacy of oral 2-methoxyestradiol capsules in hormone refractory prostate cancer.Clin Cancer Res, 2005,18(11): 6625-6633.
[33] Goud H. Desai, Susan R. Mallery, Steven P. Schwendeman. Effect of formulation parameters on 2-methoxyestradiol release from injectable cylindrical poly(DL-lactide-co-glycolide) implants. European Journal of Pharmaceutics and Biopharmaceutics, 2008, 70(12) :187-198.
[34] MacFie J. The development of fat emulsions [J]. Nut rition ,1999 , 728 (15) :641.
[35] A.Akkar , R.H.Miiller. Formulation of intravenous Carbamazepine emulsions by SolEmuls technology. European Journal of Pharmaceutics and Biopharmaceutics, 2003,234(55): 305-312.
[36] Regina G. Kelmann, Gislaine Kuminek, Helder F. Carbamazepine parenteral nanoemulsions prepared by spontaneous emulsification process. International Journal of Pharmaceutics, 2007, 342(2): 231-239.
[37] Jorge Medina, Angeles Salvado , Alfonso del Pozo. Use of ultrasound to prepare lipid emulsions of lorazepam for intravenous injection. International Journal of Pharmaceutics, 2001, 216(12): 1-8.
[38] Wang H, Yang G, Liang WQ. Preparation of the dry emulsion of bupivicaine hydrochloride [J]. Strait Pharmaceutical Journal, 2004, 16(4): 21-22.
[39] Lu YT, Dong Y, Gao CZ.. Clinical efficiency of bruceolic oil emulsion on curing liver cancer of advanced stage [J]. Modern Preventive Medicine, 2005, 32(7): 845-846.
[40]吴玉波,张永恒,孙世中等.喜树碱精氨酸静脉乳剂药物动力学及组织分布的研究[J].中国医院药学杂志,1994,14(9):406-408.
[41]高晓黎,毕殿洲,孙殿甲,等,去氢骆驼蓬碱静脉注射乳剂在动物体内的药代动力学[J].药学学报,2000,35(3):224-227
[42]高晓黎,程利勇,孙殿甲,等.定量评价去氢骆驼蓬碱注射用乳剂的组织靶向[J].药学学报,2000,35(2):142-145.
[43]陈瑞珠,汪国玮,马晋隆,等.蒿甲醚靶向性静脉乳剂的研究[J].上海医药,1997,23(1):28.
[44]Tian X,Zhu JB,Wang JS.Preparation of tetrandrine lipid emulsion and its therapeutic effect on bleomycin-induced pulmonary fibro sis in Rats[J].China Pharma Univ,2005,36(3):225-229.
[45]Jiang WS.Preparation of allicin lipid emulsion[P].CN:1579375,2005-02-16.
[46]Guo T,Chu XQ,Song HT,et al.Preparation of garlic oil submicron emulsion for intravenous administration[J].Chinese Journal of Pharmaceuticals,2005,40(8):602-605.
[47]Constantinides PP,Lambert KJ,Tustian AK,et al.Formulation development and antitumor activity of a filter-sterilizable emulsion of Paclitaxe[J].Pharm Res,2000,17(2):175-182.
[48]Adwoa O.Nornoo,David W.Osborne,Diana S.-L.Chow.Cremophor-free intravenous microemulsions for paclitaxel:Formulation,cytotoxicity and hemolysis.International Journal of Pharmaceutics,2008,349(2):108-116.
[49]Raul C.Maranh(?)o,Elaine R.Tavares,Amanda F.Padoveze.Paclitaxel associated with cholesterol-rich nanoemulsions promotes atherosclerosis regression in the rabbit.Atherosclerosis,2008,197(2):959-966.
[50]Akkar A,Muller RH.Intravenous itraconazole emulsions produced by SolEmuls technology[J].Eur J Pharm Biopharm,2003,56(1):29-36.
[51]Brime B,Moreno MA,Frutos G,et al.Amphotericin B in oil-water lecithin-based microemulsions:formulation ang toxicity evaluation[J].J Pharm Sci,2002,91(4):1178-1185.
[52]Wang Y,Cory AL.A novel stable supersaturated submicron lipid emulsion of tirilazad[J].Pharm Dev Technol,1999,4(3):333-345.
[53]Jumaa M,Muller BW.Development of a novel parenteral formulation for tetrazepam using a lipid emulsion[J].Drug Dev Ind Pharm,2001,27(10):1115-1121.
[54]Yan Lu,YanJiao Wang,Xing Tang.Formulation and thermal sterile stability of a less painful intravenous clarithromycin emulsion containing vitamin E.International Journal of Pharmaceutics,2008,346(2):47-56.
[55]Yan Lu,Yu Zhang,Ziyi Yang,et al.Formulation of an intravenous emulsion loaded with a clarithromycin-phospholipid complex and its pharmacokinetics in rats.International Journal of Pharmaceutics,2009,366(2):160-169.
[56]刘琳琳.氟尿嘧啶乳剂研制与临床应用.临床药学,2005,23(2):4
[57]T.P.Formariz,V.H.V.Sarmento,A.A.Silva-Junior,et al.Doxorubicin biocompatible O/W microemulsion stabilized by mixed surfactant containing soya phosphatidylcholine.Colloids and Surfaces B:Biointerfaces,2006,51(1):54-57.
[58]Shaimaa S.Ibrahim,Gehanne A.S.Awad,Ahmed Geneidi,et al.Comparative effects of different cosurfactants on sterile prednisolone acetate ocular submicron emulsions stability and release.Colloids and Surfaces B:Biointerfaces,2009,69(2):225-231.
[59]Panayiotis P,Alex Tustian,Dean R.Tocol emulsions for drug solubilization and parenteral delivery.Advanced Drug Delivery Reviews,2004,254(56):1243-1255.
[60]Feng Hung,Lang Fang,Mei-Hui Liao,et al.The effect of oil components on the physicochemical properties and drug delivery of emulsions:Tocol emulsion versus lipid emulsion.International Journal of Pharmaceutics,2007,335(2):193-202.
[61]Jie Li,Shufang Nie,Xingang Yang,et al.Optimization of tocol emulsions for the intravenous delivery of clarithromycin.International Journal of Pharmaceutics,2008,356(2):282-299.
[62]Shuai Shi,Hao Chen,Yue Cui,et al.Formulation,stability and degradation kinetics of intravenous cinnarizine lipid emulsion..International Journal of Pharmaceutics,2009,345(2):240-245.
[63]Archie J.Sirianni,Kevin C.Osterhoudt,Diane P.Calello,et al.Use of Lipid Emulsion in the Resuscitation of a Patient With Prolonged Cardiovascular Collapse After Overdose of Bupropion and Lamotrigine. Annals of Emergency Medicine, 2008,51(4): 412-415.
[64] Srinivas Ganta, James W. Paxton, Bruce C. Baguley, et al. Pharmacokinetics and pharmacodynamics of chlorambucil delivered in parenteral emulsion. International Journal of Pharmaceutics, 2008, 360(2): 115-121.
[65]Mukesh Kumar, Ambikanandan Misra, A.K. Babbar, et al. Intranasal nanoemulsion based brain targeting drug delivery system of risperidone. International Journal of Pharmaceutics, 2008, 358(2): 285-297.
[66] Joung Wang, K.C. Sung, Hui Yeh, Jia-You Fang. The delivery and antinociceptive effects of morphine and its ester prodrugs from lipid emulsions. International Journal of Pharmaceutics, 2008, 353(2): 95-104.
[67] Joung Wang, K.C. Sung, Oliver Yoa-Pu Hu, Hui Yeh. .Submicron lipid emulsion as a drug delivery system for nalbuphine and its prodrugs. Journal of Controlled Release, 2006,115(2): 140-149.
[68] Granato D, Blum S, Rossle C, et al, Effects of parenteral lipid emulsions with different fatty acid composition on immune cell functions in vitro[J]. JPEN J Parenter Nutr, 2002,24(2): 113-118.
[69] Nordenstrom J, Thorne A, Olivercrona A. Metabolic effects of infusion of a structured-triglyceride emulsion in healthy subjects [J]. Nurtrition,1995,11(3):269-274.
[70] Kawaguchi, Shimokawa, Fumiyoshi. Physicochemical properties of structured phosphatidylcholine in drug carrier lipid emulsions for drug delivery systems. Colloids and Surfaces B: Biointerfaces, 2008, 62(1): 130-135.
[71] M. Wulff-P(?)rez, A. Torcello-Gomez, M.J. G(?)lvez-Ruiz.. Stability of emulsions for parenteral feeding: Preparation and characterization of o/w nanoemulsions with natural oils and Pluronic f68 as surfactant. Food Hydrocolloids, 2009, 23 (4): 1096-1102.
[72] Michele Trotta, Franco Pattarino, Terenzio Ignoni. Stability of durg-carrier emulsions containing phosphatidylcholine mixtures. European Journal of Pharmaceutics and Biopharmaceutics, 2002, 53(5): 203-208.
[73] I. Cuellar, J. Bullon, A.M.Forgarini. More efficient preparation of parenteral emulsions or how to improve a pharmaceutical recipe by formulation.Chemical Engineering Science,2005,60(7):2127-2134.
[74]Seid Mahdi Jafari,Yinghe He,Bhesh Bhandari.Production of sub-micron emulsions by ultrasound and microfluidization techniques.Journal of Food Engineering,2007,82(4):478-488.
[75]R.H.Muller,S.Schmidt,I.Buttle,et al.SolEmuls-novel technology for the formulation of i.v.Emulsions with poorly soluble drugs.International Journal of Pharmaceutics,2004,269(2):293-302.
[76]陆彬.药物新剂型与新技术.北京:人民卫生出版社,2005年版.
[77]国家药典委员会.中华人民共和国药典.化学工业出版社,2005年版二部.
[78]《化学药物急性毒性研究技术指导原则》课题研究组.化学药物急性毒性研究技术指导原则.2004年版.
[79]《化学药物刺激性、过敏性和溶血性研究技术指导原则》课题研究组.化学药物刺激性、过敏性和溶血性研究技术指导原则.2004年版.
[80]司徒振强.《细胞培养技术》.世界图书出版西安公司,2007年版.