紫杉素磁靶向脂质体的研制及其抗肿瘤作用的研究

 目的：制备紫杉素磁性脂质体，考察有关理化性质、在小鼠体内的分布及其抑瘤作用。方法：用逆相蒸发法制备紫杉素磁性脂质体，对其包封率、粒径和pH值等性质进行了研究，用高效液相色谱法测定了在兔体内的药动学参数，测定了其对小鼠EMT-6肿瘤的抑瘤率。结果：紫杉素磁性脂质体包封率为97.25%,平均粒径为526 nm, pH值为6.20。结论：制备得到的紫杉素磁性脂质体适宜于非胃肠道给药，且具有明显的抑瘤作用。     

pH敏感前体阳离子脂质体的制备及瘤内分布研究

 目的 以羟喜树碱作为模型药物研究pH敏感前体阳离子脂质体用于包载抗癌药物的制备工艺并考察其pH敏感性及荷瘤鼠体内药动学特性。方法 采用薄膜分散法制备羟喜树碱阳离子脂质体;用羧甲基壳聚糖物理修饰阳离子脂质体;分离游离药物后测定包封率;用激光粒度分析仪测定Zeta电位、粒径大小;采用鸡红细胞溶血实验分析其pH敏感性并考察不同时刻药物在荷瘤小鼠血浆及瘤内浓度。结果 平均包封率为(78.5±0.9)％(n=3);Zeta电位为-31.3 mV,平均粒径为96 nm。溶血实验显示pH敏感前体阳离子脂质体与红细胞膜融合作用有明显的pH值敏感性,荷瘤小鼠体内实验显示pH敏感前体阳离子脂质体瘤内分布较高。结论 本方法制备的pH敏感前体阳离子脂质体作用机制独特,瘤内浓度高,有望成为靶向肿瘤药物的传递系统。
     

pH/还原敏感型多柔比星前药胶束的制备及体外性能评价

目的 本实验以聚乙二醇 (polyethylene glycol,PEG) 和多柔比星(doxorubicin, DOX)合成的刺激敏感型前药聚合物(PEG-DOX)包载美法仑(melphalan,MEL),制备MEL/PEG-DOX纳米胶束,考察其体外协同抗肿瘤作用。方法 通过希夫碱反应将PEG化的二硫代二丙酸二酰肼(TPH)与DOX结合,形成pH/还原敏感型PEG-DOX前药纳米载体,通过其自组装性能制备MEL/PEG-DOX载药胶束。用透射电镜观察其形态,粒径仪测定其粒径和电位,用超滤法测定载药量和包封率,用透析法评价胶束的药物释放,采用MTT法评价细胞毒性。结果 通过核磁共振氢谱验证了所制备的刺激响应型PEG-DOX前药聚合物;其PEG-DOX载体平均粒径为(188±2.4)nm,多分散系数(PDI)为(0.255±0.008);MEL/PEG-DOX载药胶束的平均粒径为(299.7±2.4)nm,多分散系数(PDI)为(0.301±0.03),Zeta电位为(-0.385±0.02) mV;DOX载药量为(14.85±0.24)%,包封率是(85.78±0.37)%,MEL载药量为(7.36±0.36)%,包封率为(38.79±0.42)%;体外药物释放实验结果表明,MEL/PEG-DOX胶束具有还原敏感和pH敏感性,且还原敏感性大于pH敏感性;细胞毒性实验分析,DOX和MEL在细胞内共同释放,实现了对肿瘤细胞的联合杀伤。结论 MEL/PEG-DOX可以在肿瘤微环境特异性释放,对肿瘤细胞具有协调治疗的作用,具有良好的应用前景。     

考布他汀4A磷酸酯缓释纳米粒的制备及抗肿瘤活性考察

目的:以聚乳酸-聚羟基乙酸共聚物(PLGA)为载体材料,制备考布他汀4A磷酸酯(CA4P)缓释纳米粒,并考察其体内外抗肿瘤活性.方法:采用W/O/W复乳-溶剂挥发法制备CA4P-PLGA纳米粒(CA4P-PLGA-NPs),通过透射电镜观察其形态,激光光散射仪测定粒径分布,HPLC测定载药量、包封率和体外累积释放率.采用MTT测定体外细胞毒活性,通过荷瘤裸鼠试验评价CA4P-PLGA-NPs体内抑瘤活性.结果:CA4P-PLGA-NPs粒径分布较窄,平均粒径113.7 nm,平均载药量(12.5±0.8)％,平均包封率(58.9±1.2)％,具有一定缓释效果.MTT和荷瘤裸鼠试验表明CA4P-PLGA-NPs具有较好的抗肿瘤活性.结论:采用W/O/W复乳-溶剂挥发法制备的CA4P-PLGA-NPs具有一定的缓释作用,抗肿瘤活性较CA4P有所提高.     

紫杉醇棕榈酸酯白蛋白纳米粒的制备及药效和骨髓抑制毒性评价

目的 制备紫杉醇棕榈酸酯白蛋白纳米粒(Nab-PTX-PA)冻干粉,对其抑瘤效果和骨髓抑制毒性进行评价。方法 采用纳米颗粒白蛋白结合技术(Nab^TM技术)制备Nab-PTX-PA冻干粉,对Nab-PTX-PA处方的形态、粒径和电位进行表征并比较Nab-PTX-PA冻干前和冻干复溶后粒径、电位的变化。通过构建4T1荷瘤小鼠模型,考察Nab-PTX-PA的抗肿瘤活性。荷瘤小鼠药效给药结束后取血进行血常规检测,考察该制剂的骨髓抑制毒性。结果 本研究成功制备了外观平整饱满、生理盐水复溶后再分散性良好的Nab-PTX-PA冻干粉,且制备Nab-PTX-PA冻干粉粒径为(87.63±1.15)nm(n=3),Zeta电位为(-11.7±0.61) mV(n=3),冻干前和复溶后Nab-PTX-PA的粒径、电位均无明显变化。药效学研究结果表明,Nab-PTX-PA(51.16 mg·kg^-1)的抗肿瘤作用高于Abraxane^® (20 mg·kg^-1),且具有统计学意义。骨髓抑制毒性结果表明,Nab-PTX-PA(25.58 mg·kg^-1)对小鼠的骨髓抑制毒性低于等剂量市售药物Abraxane^® ,Nab-PTX-PA(51.16 mg·kg^-1)对小鼠的骨髓抑制毒性与市售药物Abraxane^® (20 mg·kg^-1)相当。结论 采用Nab^TM技术制备的Nab-PTX-PA冻干粉性质稳定,Nab-PTX-PA(25.58 mg·kg^-1)与等剂量的Abraxane^® (20 mg·kg^-1)相比,降低了骨髓抑制毒性,Nab-PTX-PA(51.16 mg·kg^-1)和Abraxane^® (20 mg·kg^-1)毒性相同时,抗肿瘤效果较为明显。     

玻璃体注射用秦皮甲素微球的处方及制备工艺优化

目的 研究玻璃体注射用秦皮甲素微球的制备工艺并对其进行优化。方法 以秦皮甲素为模型药物,壳聚糖为载体材料,采用乳化交联法制备秦皮甲素微球,在单因素考察的基础上,通过星点设计进行处方工艺优化,从而得到秦皮甲素微球最佳制备条件。结果 制备所得秦皮甲素微球呈淡黄色粉末状态,外观圆整,能均匀分散于5 mg·mL^-1的羧甲基纤维素钠溶液中,其载药量为8.03%,包封率为93.03%,平均粒径为4.81 μm。结论 本研究制备得到的秦皮甲素微球具有较佳的载药量和包封率,且粒径符合玻璃体注射要求,可为黄斑病变的治疗提供新的剂型。     

苦参总生物碱脂质体凝胶的制备及释药机制研究

目的优化苦参总生物碱脂质体的处方与工艺,制备苦参总生物碱脂质体凝胶并研究其释放机制。方法运用薄膜分散法制备苦参总生物碱脂质体,以包封率和载药量为考察指标,采用酸性染料比色法测定脂质体中苦参碱含量。采用正交试验设计对处方工艺进行优化,选出最优处方制备脂质体。以泊洛沙姆-407为基质,制备苦参总生物碱脂质体凝胶。考察药物凝胶与药物脂质体凝胶的透皮速率。结果通过优化处方与工艺所得的脂质体形态均匀,粒径范围为100~400 nm,包封率达74%,载药量达26%。所制备的脂质体凝胶呈透明状半固体。体外释放结果显示,苦参碱凝胶48 h内累积释药量为6.34 mg/cm~2,苦参总生物碱脂质体凝胶48 h内累积释药量为6.97 mg/cm~2,后者的累积透皮量、稳态透皮速率及48 h后在皮肤中的蓄积量均较前者显著提高。结论优化的制备工艺稳定可行,制得的苦参总生物碱脂质体凝胶质量较好,可有效延缓药物的释放速率,提高药物在体内的蓄积量。     

线粒体靶向性的姜黄素TPP-PEG-PLGA脂质体的制备及促肝癌细胞凋亡研究

目的 研究姜黄素3-苯羧基-三苯基溴化膦-聚乙二醇-聚乳酸聚乙醇酸共聚物(TPP-PEG-PLGA)脂质体的线粒体靶向性及促肝癌细胞凋亡效果。方法 以薄膜水化法制备姜黄素TPP-PEG-PLGA脂质体,评价该载药系统的稳定性、溶血、体外释放及体内代谢情况;以荧光实验研究脂质体的肝癌细胞摄取、线粒体靶向和肝肿瘤组织靶向效果;在等剂量给药条件下,评价姜黄素TPP-PEG-PLGA脂质体促肝癌细胞凋亡效果。结果 姜黄素TPP-PEG-PLGA脂质体呈圆球型结构,具有较好的稳定性、较小的溶血率和良好的缓释药物性能;药动学结果显示,姜黄素TPP-PEG-PLGA脂质体能将药物的半衰期延长3倍,生物利用度提高4倍;荧光实验显示,TPP阳离子能促进脂质体的细胞摄取,靶向进入线粒体,还可以提高药物在肝肿瘤组织部位的靶向和滞留效果;细胞药效显示,姜黄素TPP-PEG-PLGA脂质体能明显升高肿瘤细胞内H₂O₂水平,降低线粒体膜电位,提高促凋亡蛋白Bax表达,明显降低抗凋亡蛋白BCl-2表达,这些细胞凋亡试验结果均明显优于姜黄素脂质体和姜黄素。结论 本研究表明姜黄素TPP-PEG-PLGA脂质体具有良好的线粒体靶向功能,能增强药物促肝癌细胞凋亡效果。     

阿霉素-槲皮素复方脂质体的制备

目的：研究阿霉素-槲皮素复方脂质体（DQ-Lip）的制备工艺。方法：以氢化大豆磷脂（HSPC）和胆固醇（CH）为膜材,薄膜超声结合硫酸铵梯度法制备阿霉素（DOX）-槲皮素（QUE）复方脂质体。单因素考察药脂比、硫酸铵浓度和孵育温度,优化处方工艺,超速离心后HPLC测定脂质体中DOX与QUE包封率;动态光散射粒径仪及透射电镜测定脂质体粒径及形态。结果：优化处方工艺为：QUE：HSPC=1∶20mol/mol,DOX∶HSPC=1∶7.9w/w,硫酸铵浓度为0.15mol/L,孵育温度55℃;制备得到的复方脂质体DOX、QUE的包封率分别为（90.6±0.61）%、（83.3±0.24）%（n=3）,粒径为138.5nm。结论：所得优化工艺制备阿霉素-槲皮素复方脂质体稳定可行。     

两亲性纳米胶束聚乙二醇接枝壳聚糖偶联脱氧胆酸-姜黄素的制备与评价

目的 合成聚乙二醇接枝壳聚糖偶联脱氧胆酸(mPEG-CS-DA)纳米载药系统,并以姜黄素(Cur)为模型药物,构建两亲性纳米胶束,并对其进行理化表征。方法 采用两步反应合成mPEG-CS-DA,分别用IR、¹H-NMR等技术对其结构进行表征,用差示扫描量热法(DSC)对其物理性质进行分析,并考察其在不同溶剂中的溶解性能。筛选mPEG-CS-DA-Cur纳米胶束制备方法,并测定其包封率、载药量、粒径分布及体外释放度。结果 以透析法制备的载药纳米胶束具有较高的包封率,较小的平均粒径和无残留溶剂的特点。mPEG-CS-DA-Cur的载药量为(12.11±1.52)%,包封率为(89.37±4.12)%,平均粒径为161.4 nm,分布均匀,胶束中药物体外释放缓慢、稳定。结论 研究合成了1种新型的两亲性壳聚糖衍生物,是1种良好的胶束载体材料,对难溶性药物Cur具有很好的包封率和载药量,为后续药物进行靶向性研究提供了一定的基础。     

伊马替尼胶束的制备与体外克服肿瘤耐药性的研究

目的 伊马替尼(imatinib,IMN)包埋于托可索仑(tocofersolan,TPGS)胶束中,以提高其水溶性,并对其理化性质进行表征,及与人源耐药细胞株MCF-7/ADR的作用效果。通过TPGS胶束运输IMN进入细胞,可克服肿瘤细胞的多药耐药性,提高抗肿瘤效果,为进一步体内药效研究提供基础。方法 采用薄膜分散法制备IMN-TPGS胶束,对其粒径、电位、形态、载药量和包封率及体外释放行为进行表征,考察该胶束对MCF-7/ADR细胞的抗肿瘤效果。结果 IMN-TPGS胶束平均粒径为(22.69±2.39)nm,表面呈电中性,外观圆整,载药量为(1.55±0.06)%,包封率为(63.49±2.42)%;具有较高的耐稀释性,血清稳定性和冻干-复溶稳定性;在体外释放72 h后,累积释放度近100%,释放速率缓慢;IMN-TPGS胶束对MCF-7/ADR的IC₅₀为12.27 μmol·L^-1,细胞毒性大,细胞内药物含量高,可诱导近半数细胞凋亡。结论 IMN-TPGS胶束粒径均一,分散性良好,稳定性高,具有较强抗稀释性,有利于体液长循环,对耐药细胞株有明显的抗肿瘤效果,在一定程度上可以克服其多药耐药性。     

小檗碱磷脂固体分散体改善氧化应激治疗2型糖尿病

目的 制备包载小檗碱(BBR)的磷脂固体分散体促进药物吸收,增强降糖、降脂药效,并初步阐明其抗2型糖尿病(T2DM)作用机制.方法 溶剂挥发法制备小檗碱磷脂固体分散体(SD),对其粒径、形貌、体外释放、胃肠道吸收、治疗T2DM药效和作用机制进行了充分研究.结果 SD在水溶液中粒径约为118 nm,形貌为球形;相比于游离...     

主动靶向型载奥拉帕尼PEI-PLGA纳米粒的制备表征及抗三阴乳腺癌研究

目的 构建具有肿瘤细胞特异靶向性的纳米给药系统,研究该纳米粒的体外抗三阴乳腺癌作用。方法 采用乳化扩散溶剂挥发法制备包载奥拉帕尼的聚乙烯亚胺-聚乳酸-羟基乙酸共聚物(PEI-PLGA)纳米粒,进一步用透明质酸修饰,制备靶向载药纳米粒。采用激光粒度仪和透射电镜对纳米粒的形态、粒径大小、电位及分布情况进行表征。CCK-8法检测纳米粒对MDA-MB-231细胞的毒性作用,通过共聚焦显微镜观察细胞对目标纳米粒的靶向摄取,生物透射电镜观察细胞内部形态变化分析凋亡情况。结果 成功制备奥拉帕尼靶向载药纳米粒(HA-Ola-PPNPs),粒径为(166.2±0.842) nm,大小分布均匀,呈圆形或椭圆形,稳定性良好;奥拉帕尼包载于PEI-PLGA纳米粒中能够起到药物缓释作用;CCK-8实验结果显示纳米粒组均呈现出时间依赖和剂量依赖性细胞毒性,且HA-Ola-PPNPs对癌细胞具有更强的杀伤作用;激光共聚焦结果说明了HA与CD44受体的主动靶向结合可以显著提高MDA-MB-231细胞对目标纳米粒的摄取;生物透射电镜观察显示HA-Ola-PPNPs能够有效诱导MDA-MB-231细胞发生凋亡。结论 本研究成功制备了具有CD44受体特异性靶向的载药纳米粒,为三阴乳腺癌的临床治疗提供一种新策略。     

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??OBJECTIVE To prepare the transferrin modified harmine magnetoliposomes and investigate its in vitro properties and antitumor effect. METHODS Transferrin modified harmine magnetoliposomes was prepared by reverse phase evaporation method. The size, Zeta potential, entrapment efficiency, drug loading efficiency, release rate and plasma stability of liposomes were investigated. Furthermore, cytotoxicity to liver cancer and glioma were investigated too. RESULTS The transferrin modified harmine magnetoliposomes were successfully prepared. The average size of liposomes was (184.7??8.08)nm.The average Zeta potential was (-12.4??0.896) mV and average entrapment efficiency was (84.67??6.26)%. Drug loading efficiency was (9.14??4.54)%. Transferrin modified harmine magnetoliposomes released more slowly and were more stable in rat plasma than harmine solution. The results of cytotoxicity test showed that the cell inhibitory effect of transferrin modified harmine magnetoliposomes was enhanced compared with other groups, and the cell inhibitory effect of this preparation was more obvious underwhen cells were put under magnetic field compared with those under the non-magnetic field(P<0.05). CONCLUSION This method can be successfully used to prepared transferrin modified harmine magnetoliposomes, and liposomes can inhibit the cell visbility of liver cancer and human glioma cells significantly.     

巨噬细胞膜仿生白蛋白纳米体系的构建及其胶质瘤靶向递药性能评价

目的 构建包载WEE1激酶抑制剂adavosertib的巨噬细胞膜仿生白蛋白纳米粒(MM-BSA/Ada),体外评估其作为胶质瘤靶向递药体系的可行性。方法 制备MM-BSA/Ada并筛选最佳膜-核比和最佳药-载比,检测其载体安全性和对C6胶质瘤细胞抗增殖活性,考察其体外细胞摄取、跨血脑屏障转运和跨膜后摄取的能力。结果 MM-BSA/Ada具有良好的稳定性,CCK-8结果初步显示,未载药纳米粒在体外细胞实验中对脑血管内皮细胞呈低毒性;与未包膜纳米粒和游离药物相比,MM-BSA/Ada给药后的体外抗胶质瘤细胞增殖活性(P＜0.001)、胶质瘤细胞摄取量(P＜0.001)、体外血脑屏障透过量(P＜0.01)及跨膜后摄取量(P＜0.001)均显著提高。结论 MM-BSA/Ada有较好的胶质瘤靶向递药性能,有望为胶质瘤提供新的放射增敏策略。     

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??OBJECTIVE To prepare calcitonin/puerarin-PLGA-dual-loaded nanoparticles modified by chitosan, and investigate theirin vitro release behavior.METHODS CS-CT/PR-NPs were prepared by the double emulsion solvent evaporation technique with PLGA as a carrier material; the formulation of CS-CT/PR-NPs was optimized by orthogonal design; the morphology of CS-CT/PR-NPs was observed by transmission electron microscope;the mean particle size,particle size distribution and Zeta potential were measured by laser particle size analyzer; the entrapment efficiency and drug loading were measured by ultracentrifugation; the in vitro release behavior was studied by dialysis. RESULTS CS-CT/PR-NPs were spherical in shape with the mean particle size of(190??2.65) nm, particle size distribution of (0.117??0.027) and Zeta potentialof(16.5??1.08) mV. The entrapment efficiency was (75.7??1.15)%, and the drug loading of CT was (3.47??0.31)%, while those of PR were (50.9??1.08)% and (4.68??0.19)%, respectively. The profiles of in vitro release had the features of sustained-release. CONCLUSION CS-CT/PR-NPs are prepared successfully and show a sustained-release characteristic with high entrapment efficiency, which may improve the oral bioavailability of CT and provide the experimental reference for preparing the dual-loaded nanoparticles.     

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??OBJECTIVE To prepare nano-micelles with amphiphilic self-assembly poly (ethylene glycol)-co-poly (propylene sulfide) (PEG-PPS) copolymer as carrier to study the release characteristics of tilianin and investigate its activity to against H9c2 cell apoptosis in vitro. METHODS An amphiphilic diblock PEG-PPS polymer was used as a carrier material to prepare the tilianin-containing nano-micelles by solvent evaporation. The morphology, particle size and distribution, drug loading and encapsulation rate and in vitro drug release behavior were characterized, H9c2 rat myocardial cell injury model was established by hypoxia/reoxygenation process. Using propranolol (Pro) as a positive control, the morphology of injured cardiomyocytes was observed by microscope. Cell proliferation and cell apoptosis was detected to evaluate the protective effect of blank micelles, tilianin and tilianin loaded nano-micelles on H9c2 cells induced by hypoxia/reoxygenation. RESULTS Tilianin-loaded nano-micelles was spherical with uniform particle size distribution. The drug loading was 3.82%. The average particle diameter of tilianin-loaded nano-micelles was 137 nm, polydispersity coefficient was 0.162 and the encapsulation efficiency was 91.45%. In vitro drug release studies showed that there was no drug-induced burst release of tilianin-containing nano-micelles and sustained release characteristics, and the presence of hydrogen peroxide significantly promoted the release of tilianin from the nano-micelles. In vitro cytotoxicity experiments showed that when the concentration of tilianin 5 ??g??mL^-1, the cell viability of tilianin-loaded nano-micelles was significantly higher than the corresponding concentration of tilianin and PEG-PPS polymer nano-micelles. In vitro anti-apoptotic activity experiments show that tilianin-loaded nano-micelles on H9c2 cell apoptosis induced by hypoxia-reoxygenation have a significant inhibitory effect and was provided inhibition of apoptosis with propranolol. CONCLUSION Tilianin-loaded nano-micelles have uniform particle size and distribution, sustained release and oxidation characteristics, has a significant protective and apoptosis-inhibiting effect on H9c2 cell injury induced by hypoxia-reoxygenation, which can be used as a promising drug delivery system for the treatment of myocardial ischemia-reperfusion injury.