汉黄芩素固体脂质纳米粒的制备及体外释放度考察

目的:制备汉黄芩素固体脂质纳米粒并对其体外释放度进行考察.方法:采用乳化分散-超声法制备汉黄芩素固体脂质纳米粒,以包封率和载药量为评价指标,进行正交试验筛选最优处方,并对最优处方的外观、粒径和体外释放度进行考察.结果:制得的纳米粒为均一球形,平均粒径为(153 ±34)nm,其平均载药量为(60.53±2.17)％,平均包封率为(85.54±4.16)％,48 h累积释放达80％.结论:本试验获得了较理想的汉黄芩素固体脂质纳米粒,其体外释放具有缓释作用.     

紫杉醇长循环固态脂质纳米粒的制备和体内外研究

目的以硬脂酸为载体材料制备紫杉醇的长循环脂质纳米粒,并考察其体内外性质。方法用“乳化蒸发-低温固化”法制备Brij78固态脂质纳米粒(Brij78-SLN)和Poluromic F₆₈固态脂质纳米粒(F₆₈-SLN);用透射电镜考察了紫杉醇纳米粒的形态;建立了脂质纳米粒和血清中测定紫杉醇的HPLC方法;考察了纳米粒于30％乙醇溶液中的体外药物释放;以市售紫杉醇注射剂对照,测定了两种纳米粒于小鼠体内的药物动力学参数。结果脂质纳米粒基本呈圆球状或椭圆球状,大小比较均匀。激光散射法测定Brij78-SLN粒径为(104±29) nm。F₆₈-SLN粒径为(220±98) nm。Brij78-SLN和F₆₈-SLN包封率分别为47%和75%。两种纳米粒都缓慢地释放药物,24 h后分别释放药物总量的8%和20%。两种纳米粒都可以延长紫杉醇的体内滞留时间,Brij78-SLN,F₆₈-SLN和紫杉醇注射剂的消除半衰期分别为4.88,10.06和1.36 h。结论硬脂酸纳米粒可能成为一种新型的药物载体。     

青蒿琥酯纳米粒的制备及其体外细胞抑制试验

目的:制备青蒿琥酯纳米粒,并对其性质及体外细胞抑制作用进行研究。方法:以聚乳酸-羟基乙酸共聚物(PLGA)为载体,采用自乳化方法制备青蒿琥酯纳米粒。扫描电镜观察纳米粒的形态,激光粒度仪测定纳米粒的粒径及其分布;考察纳米粒的载药量、包封率、体外释放情况;MTT法考察纳米粒对人白血病细胞株K562在不同时间(24、48、72h)的体外细胞抑制率,并与青蒿琥酯(原料药)比较。结果:所制青蒿琥酯纳米粒为圆球形,表面光滑,平均粒径为(144±3.0)nm,Zeta电位是-31.5mV,平均载药量和包封率分别为14%、84%;体外释放试验前期有明显突释现象,前24h累积释放度为46%,其后释放均匀,120h累积释放度达65%,具有缓释作用;其在72h时对细胞抑制率高于青蒿琥酯组(76.4%vs.59.1%),有较强抑制作用(P<0.05)。结论:所制青蒿琥酯纳米粒在体外具有较好的缓释性,对K562细胞有较强的抑制作用。     

不同粒径多西他赛固体脂质纳米粒制剂及其细胞毒活性

目的制备不同粒径的多西他赛(docetaxel,DTX)固体脂质纳米粒,考察多西他赛固体脂质纳米粒理化性质,研究粒径对体外释放行为以及细胞毒作用的影响。方法通过热熔超声法制备不同粒径多西他赛固体脂质纳米粒,观察纳米粒形态,测定其包封率、粒径、Zeta电位。考察粒径因素对固体脂质纳米粒体外释放行为、体外细胞毒性的影响。结果制备的纳米粒均为球形及类球形,3种粒径的多西他赛固体脂质纳米粒平均粒径分别为(83.7±8.4)、(162.7±11.9)、(232.1±26.4)nm;Zeta电位分别为-24.19、-23.67、-23.19 mV;包封率分别为98.03%、97.84%、97.92%。60 h粒径分别为83、16、232 nm的多西他赛固体脂质纳米粒在释放介质中分别累计释放86.34%、76.98%、67.14%。3种不同粒径多西他赛固体脂质纳米粒(DTX-SLN-83,DTX-SLN-162,DTX-SLN-232),多西他赛原料药(DTX solutions)以及空白SLN溶液与多西他赛的混合溶液(physi-calm ixture)对MCF-7细胞作用24 h的IC50值分别为3.36、6.20、9.74、13.15、12.92 mg.L-1;48 h的IC50值分别为0.93、2.01、4.35、9.48、9.21 mg.L-1;72 h的IC50值分别为0.30、0.91、1.67、7.36、7.82 mg.L-1。随着多西他赛固体脂质纳米粒粒径的减小,其肿瘤细胞杀伤力逐渐增强。结论热熔超声法可用于制备不同粒径多西他赛固体脂质纳米粒。降低固体脂质纳米粒粒径有利于药物更完全地释放,同时增强其对肿瘤细胞的杀伤能力。     

HPLC法测定槲皮素固体脂质纳米粒的含量

目的建立HPLC法测定槲皮素固体脂质纳米粒含量的方法。方法色谱柱:Diamonsil C18柱,流动相:甲醇-4.3%乙酸溶液(55∶45),流速:1.0mL.min-1,检测波长:254nm,进样量:20μL,柱温:30℃。结果槲皮素在20.0~200.0μg.mL-1范围内线性关系良好,平均回收率为98.6%,RSD=1.18%,结论本方法快捷、简便、准确、专属性强,可用于槲皮素固体脂质纳米粒的质量控制。     

自制多西紫杉醇脂质体在家兔肝组织中浓度的测定

目的:建立以固相萃取-效液相色谱法(SPE-HPLC)测定家兔肝组织匀浆中多西紫杉醇的方法,考察自制多西紫杉醇脂质体家兔耳缘静脉给药24 h后在肝组织中的浓度.方法:以地西泮为内标,家兔肝组织匀浆样品中多西紫杉醇经固相萃取后,采用大连依利特Hypersil C18(250 mm×4.6 mm,5μm)色谱分析柱,流动相为乙腈-水(47:53),流速1.0 mL·min-1,检测波长230 nm,进样量50μL.家兔耳缘静脉注射给予多西紫杉醇脂质体制剂,剂量为7.5 mg·kg-1,24 h后解剖动物,用所建立的HPIC法测定其肝组织中药物浓度.结果:地西泮和多西紫杉醇的保留时间分别是10.9 min和13.1 min,多西紫杉醇的最低检测质量浓度为5μg·L-1(S/N=3),肝组织样品中多西紫杉醇在0.025 1.5 mg·L-1范围内与峰面积比之间线性关系良好,回归方程为Y=196.42C-0.642 4,γ=0.999 2(n=7),提取回收率为81.36%～86.72%,方法回收率为91.37%～100.34%,日内RSD为2.72%,3.20%,3.55%,日间RSD为4.68%,4.47%,3.52%,且稳定性良好.实测6例多西紫杉醇脂质体在家兔肝组织中的质量浓度为0.039～0.063 mg·L-1.结论:本试验建立的方法简便灵敏,干扰小,重复性好,可实际应用于多西紫杉醇在家兔肝组织中的测定,并作为评价多西紫杉醇脂质体在靶组织分布的方法学依据.     

HPLC法测定多西他赛固体脂质纳米粒的药物含量

目的建立多西他赛固体脂质纳米粒的含量测定方法。方法采用Hypersil ODS C18柱(4.6 mm×200mm,5μm),流动相为乙腈-水(60∶40,V/V),检测波长:228 nm,流速:1 mL.min-1。结果多西他赛在0.50～50.00μg.mL-1的浓度范围内,峰面积对浓度有良好的线性关系(R2=0.999 9,n=7),方法的日内与日间精密度RSD均<2%,回收率分别为98.81%、99.22%、101.5%。结论该方法具有简便、快速、准确的特点,可用于多西他赛固体脂质纳米粒的含量测定。     

脂蟾毒配基-乳酸羟基乙酸共聚物纳米粒的制备与表征

目的:制备、表征脂蟾毒配基-乳酸羟基乙酸共聚物纳米粒(RPN)。方法:用乳酸羟基乙酸共聚物为载体材料,采用超声乳化-溶剂挥发法制备RPN,以粒径、载药量、包封率和体外释放度表征其质量。用反相-高效液相色谱法测定RPN含量和体外释放度,色谱柱为HYPERSILC18(250 mm×4.6 mm,5μm),流动相为甲醇-0.05%冰醋酸水溶液(9∶1,V/V),检测波长为298 nm。结果:RPN的平均粒径为(232.3±2.3)nm,载药量为(18.3±0.3)%,包封率为(72.3±1.2)%,体外药物呈两相释放。结论:RPN载药量较高,体外释放试验显示具有明显的缓释作用。     

人参皂苷Rg3温敏纳米粒制备及对肝癌细胞抑制作用研究

目的制备人参皂苷Rg3温敏纳米粒,对其体外释药特性和对肝癌细胞的抑制作用进行研究。方法两亲性温敏聚合物作为药物载体,滴制法制备载药温敏纳米粒。采用正交试验设计,以包封率为主要评价指标,温敏聚合物浓度、人参皂苷Rg3浓度和搅拌时间为考察因素,筛选纳米粒处方和工艺。体外释放曲线法研究37℃和42℃时温敏纳米粒的体外释放特征。MTT法(四氮唑盐微量酶反应比色法)考察温敏纳米粒对HepG2肝癌细胞增生的抑制作用。结果优选处方及制备工艺为温敏聚合物,浓度为0.20 mg/m L,人参皂苷Rg3浓度为0.03 mg/m L,搅拌时间为2 h;温敏纳米粒呈类球形,包封率为78.43%±1.76%,平均粒径为(147.0±2.3)nm,平均Zeta电位为-14.6 m V(n=3)。体外释放结果表明,37℃时,3 h药物累积释放度为36.1%,24 h累积释放度为44.6%;42℃时,3 h累积释放度达54.1%,24 h累积释放率为83.0%。体外抑制结果显示,随着温敏纳米粒浓度从20μg/m L增加至180μg/m L,HepG2肝癌细胞成活率由69.8%下降至37.7%。结论人参皂苷Rg3温敏纳米粒处方和工艺合理可行,能达到随温度变化对药物释放起调释作用,对HepG2肝癌细胞具有较好的体外抑制作用。     

紫杉醇注射液在小鼠体内的药代动力学及组织分布

目的:建立血清中紫杉醇的分析测定方法,进行紫杉醇注射液在小鼠体内的药代动力学研究。方法:采用固相萃取方法进行血清样品预处理。色谱柱为Intertex C18(150 mm×4.6 mm,5μm),流动相为磷酸水溶液(pH3.0)-乙腈(50∶50),流速为1 mL.min-1,检测波长为227 nm。结果:紫杉醇在血清和组织中的浓度范围分别为0.39~100.00μg.mL-1和0.39~25.00μg.mL-1,准确度均在85%~115%,稳定性RSD均小于15%,小鼠静脉注射紫杉醇注射液后在小鼠体内代谢符合三房室模型(W=1/C2)。结论:本方法简便快捷,灵敏准确,本文建立的紫杉醇血药浓度测定方法和紫杉醇注射液在小鼠体内的药代动力学参数,为紫杉醇其他制剂的研究提供参考。     

RP—HPLC法测定肿瘤患者血浆中紫杉醇浓度

目的：建立以反相高效液相色谱法测定肿瘤患者血浆中紫杉醇浓度的方法。方法：采用有机相（乙酸乙酯）两步萃取,以反相高效色谱法测定,其中色谱柱为Tianhe Kromasil C18,流动相为乙腈-甲醇-水（40：25：40）,流速为1．0mL·min^-1,检测波长为227nm,枉温为35℃。结果：血浆中紫杉醇的检测浓度在0．05～5．00mg·L^-1。范围内线性关系良好（r=0．9997）;平均加样回收率为98．75％～100．44％,日内、日间RSD均〈5％（n=5）。应用本法测定了11例患者静脉滴注紫杉醇的经一时血药浓度,其符合药动学二室模型。结论：本法简便、准确、重现性好,可用于临床血药浓度的检测及药动学研究。     

紫杉醇固体脂质纳米粒大鼠体内药动学

目的研究紫杉醇固体脂质纳米粒在大鼠体内的药动学。方法10只健康大鼠,雌雄各半,分为2组,分别口服给药紫杉醇固体脂质纳米粒和紫杉醇乳剂30 mg.kg-1,在设计的时间点从颈静脉取血,采用RP-HPLC测定紫杉醇在全血中的药物浓度,药动学参数用3P97软件进行处理。结果大鼠口服给药后,紫杉醇固体脂质纳米粒和乳剂的tm ax分别为3.133 h和1.627 h,MRT分别为10.362 h和3.297 h,mρax分别为1.512 2 mg.L-1和0.718 9 mg.L-1。结论固体脂质纳米粒能够显著改善大鼠体内紫杉醇的药动学行为,有利于其更好地发挥抗肿瘤作用。     

Modulating the therapeutic activity of nanoparticle delivered paclitaxel by manipulating the hydrophobicity of prodrug conjugates

A series of paclitaxel prodrugs designed for formulation in lipophilic nanoparticles are described. The hydrophobicity of paclitaxel was increased by conjugating a succession of increasingly hydrophobic lipid anchors to the drug using succinate or diglycolate cross-linkers. The prodrugs were formulated in well defined block copolymer-stabilized nanoparticles. These nanoparticles were shown to have an elimination half-life of approximately 24 h in vivo. The rate at which the prodrug was released from the nanoparticles could be controlled by adjusting the hydrophobicity of the lipid anchor, resulting in release half-lives ranging from 1 to 24 h. The diglycolate and succinate cross-linked prodrugs were 1-2 orders of magnitude less potent than paclitaxel in vitro. Nanoparticle formulations of the succinate prodrugs showed no evidence of efficacy in HT29 human colorectal tumor xenograph models. Efficacy of diglycolate prodrug nanoparticles increased as the anchor hydrophobicity increased. Long circulating diglycolate prodrug nanoparticles provided significantly enhanced therapeutic activity over commercially formulated paclitaxel at the maximum tolerated dose.     

RP-HPLC法测定藁本内酯脂质体的体外释放度

目的建立以反相高效液相色谱法测定藁本内酯脂质体的体外释放度的方法。方法采用溶出度第三法考察脂质体的体外释放过程。色谱条件为：色谱柱：ZORBAX-C18柱（4.6mm×250mm,5μm）;流动相：甲醇-乙腈-水（34：20：46）;流速1.0ml·min^-1;检测波长274nm。结果药物的体外释放以对数正态分布方程拟合最好。结论该方法简便、易操作,可作为藁本内酯脂质体体外释药的研究。     

黄芩苷固体脂质纳米粒体外释放研究

目的:研究黄芩苷固体脂质纳米粒的体外释药规律。方法:采用动态透析技术研究黄芩苷固体脂质纳米粒的体外释药性能,并用高效液相色谱法测定黄芩苷含量,以累积释药百分率进行不同模型的拟合。结果:黄芩苷固体脂质纳米粒的释放曲线符合Hixon-crowell方程,t≈3h。结论:黄芩苷固体脂质纳米粒具有良好的缓释作用。     

苦参总生物碱固体脂质纳米粒的制备工艺研究

目的 探索苦参总生物碱(TA)磷脂复合物固体脂质纳米粒(TA-PC-SLN)的制备工艺及处方优化,并对其进行体外评价.方法 建立高效液相色谱(HPLC)法同时测定TA中4个指标成分苦参碱(MT)、氧化苦参碱(OMT)、槐果碱(SC)、氧化槐果碱(OSC),并进行方法学验证.称取TA、卵磷脂共同溶于乙醇制备磷脂复合物;采...     

PLGA/TPGS Nanoparticles for Controlled Release of Paclitaxel: Effects of the Emulsifier and Drug Loading Ratio

PURPOSE: We successfully manufactured nanoparticles of biodegradable polymers for controlled release of paclitaxel. TPGS (d-alpha-tocopheryl polyethylene glycol 1000 succinate) could be a novel material to make nanoparticles of high drug encapsulation efficiency (EE) and desired physicochemical and pharmaceutical properties of the drug loaded nanoparticles. Among various controlling parameters in the process, the present work is to elucidate the effects of the surfactant stabilizer and the drug loading ratio. METHODS: Paclitaxel loaded PLGA nanoparticles were formulated at various drug-loading ratios by a modified single emulsion solvent extraction/evaporation technique. TPGS was introduced either as the emulsifier or as a matrix material component by using different technique. Polyvinyl alcohol (PVA) was also used for a comparison. The nanoparticles of various recipes were characterized by various state-of-the-art instrument technology for their properties. RESULTS: The EE and the in vitro release behavior were found significantly influenced by the drug loading ratio and the surfactant stabilizer encountered. TPGS involved nanoparticles can have high EE and other favorable properties. CONCLUSIONS: TPGS could be a novel and effective emulsifier, which can result in high EE and desired properties of paclitaxel-loaded polymeric nanoparticles.     

紫杉醇对兔血管内皮和平滑肌细胞增生影响的差异及意义

目的探讨紫杉醇对兔血管内皮和平滑肌增生影响的差异及意义.方法将兔血管平滑肌细胞接种于共培养体系上室、内皮细胞接种于下室建立体外内膜修复模型,观察紫杉醇对兔血管平滑肌和内皮细胞3H-TdR掺入、细胞计数和迁移率的影响,用直线回归法计算紫杉醇对平滑肌和内皮细胞增生迁移的半数有效抑制浓度IC50.结果在1 nmol·L-1～1 μmol·L-1之间,紫杉醇呈浓度依赖地抑制平滑肌细胞3H-TdR掺入、细胞计数和迁移(n=6, P＜0.01).在10 nmol·L-1～1 μmol·L-1之间,紫杉醇呈浓度依赖地抑制内皮细胞3H-TdR掺入、细胞计数和迁移(n=6, P＜0.01).1 nmol·L-1紫杉醇对内皮细胞3H-TdR掺入和细胞计数有抑制倾向,但与对照组相比无统计学差异.而1 nmol·L-1的紫杉醇却已显著抑制内皮细胞迁移(n=6, P＜0.01).紫杉醇对兔血管平滑肌细胞增生、迁移抑制的IC50分别为 10.09±0.47、9.16±0.54 nmol·L-1,对内皮细胞增生、迁移抑制的IC50分别为 19.05±0.35、5.37±0.51 nmol·L-1.10 nmol·L-1紫杉醇作用 20 min 在观察时间内能持续抑制融合内皮组平滑肌增生,而对数内皮组平滑肌增殖在10 d时明显高于对照组.结论紫杉醇在抑制兔血管平滑肌细胞增生的同时也抑制内皮增生,紫杉醇干预后平滑肌细胞增生延迟与内皮细胞再生延迟密切相关.     

Preparation and characterization of polymeric pH-sensitive STEALTH® nanoparticles for tumor delivery of a lipophilic prodrug of paclitaxel

Paclitaxel is an effective and widely used anti-cancer agent. However, the drug is difficult to formulate for parenteral administration because of its low water solubility and Cremophor EL, the expient used for its formulation, has been shown to cause serious side effects. The present study reports an alternative administration vehicle involving a lipophilic paclitaxel prodrug, paclitaxel oleate, incorporated in the core of a nanoparticle-based dosage form. A hydrophobic poly (β-amino ester) (PbAE) was used to formulate the nanoparticles, which were stabilized with a mixture of phosphatidylcholine, Synperonic? F 108, and poly(ethylene glycol)-dipalmitoyl phosphatidyl ethanolamine. PbAE undergoes rapid dissolution when the pH of the medium is less than 6.5 and is expected to rapidly release its content within the acidic tumor microenvironment and endo/lysosome compartments of cancer cells. PbAE nanoparticles were prepared by an ultrasonication method and characterized for particle size and physical stability. The nanoparticles obtained had a diameter of about 70 nm and a good physical stability when stored at 4 °C. In vitro cellular uptake and release of paclitaxel oleate PbAE nanoparticles were studied in Jurkat acute lymphoblastic leukemia cells. The results were compared with pclitaxel oleate in poly(?-caprolactone) (PCL) particles, that do not display pH-sensitive release behavior, and paclitaxel in PbAE particles. Both uptake and release of the prodrug were faster when administered in PbAE than in PCL, but much slower than those of the free drug in PbAE. Cytotoxicity assay was performed on the formulations at different doses. Paclitaxel and paclitaxel oleate showed almost identical activity, IC50 123 and 128 nM, respectively, while that of the prodrug in PCL was much lower with IC50 at 2.5 μM. Thus, PbAE nanoparticles with the incorporated paclitaxel prodrug paclitaxel oleate may prove useful for replacement of the toxic Cremophor EL and also by improving the distribution of the drug to the tumor.