星点设计-效应面法优化羟基喜树碱/mPEG-S-S-C18纳米粒的制备工艺

目的 制备载羟基喜树碱（hydroxycamptothecin,HCPT）还原响应mPEG-S-S-C18纳米粒,采用星点设计-效应面法筛选优化制备工艺。方法 采用乳化-溶剂挥发法制备HCPT/mPEG-S-S-C18纳米粒,应用单因素法考察投药量、水相/油相体积比、超声功率以及超声时间对载药纳米粒包封率和载药量的影响。在此基础上,以包封率和载药量作为评价指标,采用Design-Expert V8.0.6软件进行星点设计,优化载药纳米粒的制备工艺。结果 优化获得的HCPT/mPEG-S-S-C18纳米粒制备工艺投药量为1.0 mg,水相/油相体积比为4.56∶1,超声功率为562.5 W。该工艺制备的载药纳米粒包封率为（58.14±1.04）%,载药量为（3.46±0.22）%,平均粒径为（322.9±9.52） nm,多分散性指数为0.195±0.05,Zeta电位为（-17.5±2.11） mV。结论 乳化-溶剂挥发法适用于制备HCPT/mPEG-S-S-C18纳米粒,星点设计-效应面法可优化获得载药纳米粒的最佳制备工艺,所得的载药纳米粒包封率和载药量较高,所建立的数学模型预测性良好。     

星点设计-效应面法优化流化床制备盐酸普萘洛尔微囊工艺

目的优化盐酸普萘洛尔微囊的制备工艺。方法采用流化床制备普萘洛尔微囊,以平均粒径,包封率、载药量及总评归一值为评价指标,并运用星点设计考察囊材液流速、喷雾压力对制备工艺的影响,对结果进行多元线性和二项式拟合,效应面法选取最佳工艺条件进行预测分析。结果从复相关系数上看,各指标二项式拟合方程均优于多元线性回归方程,最佳工艺参数:囊材液流速1.00 mL·min-1,喷雾压力0.65 bar,在此工艺条件下得到微囊粒径为300μm,载药量为20.54%,包封率达89.63%。结论优选普萘洛尔微囊的流化床制备工艺稳定可行,包封率高,有利于工业化生产。     

CA4P脂质体的制备工艺研究

目的 研究CA4P脂质体的处方及制备工艺.方法 以包封率为主要评价指标考察制备方法;用透射电镜和粒径测定仪表征脂质体的形态和粒径;用HPLC法测定脂质体中CA4P的包封率和载药量;以正交设计筛选优化最佳处方工艺.结果 脂质体的平均粒径为167 nm,Zeta电位为-24.3 mV,最佳工艺处方的药-脂比为1:12,磷脂-胆固醇为8:1,有机相-水相体积为4:1,水合介质为0.9%NaCl;制备3批脂质体的平均包封率为58%、载药量为4.8%.结论 逆相蒸发-探头超声法可制备具有较高包封率的CA4P脂质体.     

重组人血管内皮抑制素壳聚糖纳米粒的制备

目的:通过星点设计-效应面法优化重组人血管内皮抑制素(Endostar)壳聚糖纳米粒的制备工艺。方法:以壳聚糖浓度、三聚磷酸钠(TPP)浓度及壳聚糖溶液与TPP溶液体积比为考察因素,以载药量、包封率和粒径为指标,采用多元线性回归和多元非线性回归拟合选择合适模型,并根据最佳模型绘制效应面图,选择最佳处方,并进行预测分析,同时考察最佳处方的体外释放。结果:用多元非线性回归对实验中各因素和指标进行拟合明显优于线性拟合。最佳处方制备的Endostar载药壳聚糖纳米粒,载药量、包封率及粒径的实测值与预测值的偏差均在±7%以内,体外释放1周累积释放80%。结论:通过星点设计-效应面法可以准确快速优化处方工艺。优化后的工艺制备的纳米粒可以延缓Endostar的释放,达到了缓释效果,符合预期的试验目标。     

星点设计-效应面法优化PLGA-PEG纳米粒的处方工艺

摘要：目的:采用星点设计-效应面法优化载体基质为PLGA-PEG的siRNA纳米粒的制备工艺。方法:采用复乳法制备载药纳米粒,以二氯甲烷体积、吐温-80的质量分数和复乳的超声时间为试验因素,纳米粒的平均粒径、包封率和突释量为考察指标,根据星点设计原理安排实验和处方工艺优化。结果:成功制备了纳米粒。最佳工艺为二氯甲烷体积13 ml,乳化剂吐温-80的质量分数为3.1%,复乳的超声时间为2.8 min;按优化处方工艺制备的纳米粒的平均粒径（101.5±6.3）nm,包封率（57.6±4.8）%,体外48 h累积释放度高于80%。结论:星点设计-效应面法适用于PLGA-PEG纳米粒的工艺优化,所建立的数学模型预测性良好。     

星点设计-效应面法优化美斯地浓聚乳酸纳米粒处方

目的星点设计-效应面法优化美斯地浓聚乳酸纳米粒处方。方法以复乳液中干燥法制备美斯地浓聚乳酸纳米粒,以包封率和载药量为评价指标,在单因素试验的基础上,用星点设计对显著性因素进行优化,并进行二项式方程拟合,以效应面法选取较好的工艺条件进行预测。结果以效应面法优选出的最佳工艺为:美斯地浓投药量为49.20 mg,PLA浓度为3.31%,PVA浓度为3.41%。制备的美斯地浓聚乳酸纳米粒平均包封率和载药量分别为(51.98±1.28)%和(7.01±0.31)%(n=3),与二项式拟合方程预测值相差<2%。结论应用星点设计-效应面法优化美斯地浓聚乳酸纳米粒制备工艺,能够快速、准确的得到最佳制备工艺,预测性良好。     

星点设计-效应面法优化平阳霉素脂质体的处方及工艺

目的：优化载平阳霉素脂质体的处方及其制备工艺。方法：以其包封率为考察指标,以平阳霉素与所用脂质的比、所用磷脂与胆固醇的比、制备温度为考察项,采用星点设计-效应面法优化盐酸平阳霉素脂质体的处方及工艺。结果：确定其最优处方为平阳霉素与脂质的比为0.15（W/W）,所用磷脂与胆固醇的比为4.5（W/W）,制备温度为42.3℃,实测值与预测值的最终差异显示无统计学意义。该脂质体的平均粒径为136 nm,Zeta电位为（-28.55±6.81）mV。结论：所选制备工艺较为合理,星点设计-效应面法可用于该制剂的处方优化。     

星点设计-效应面法优化转铁蛋白修饰粉防己碱与硫酸长春新碱脂质体的处方

目的:制备转铁蛋白(TF)修饰粉防己碱(TET)与硫酸长春新碱(VCR)的主动靶向脂质体,优化其处方。方法:以硫酸铵梯度法制备TF修饰TET与VCR脂质体,以TET包封率、VCR包封率的综合评分为指标,用星点设计-效应面法优化卵磷脂/胆固醇(EPC/Chol)摩尔比、卵磷脂/聚乙二醇2000-二硬脂酰磷脂酰乙醇胺(EPC/PEG2000-DSPE)摩尔比、TF质量分数,并进行验证试验。结果:最优处方为EPC/Chol摩尔比1.5∶1,EPC/PEG2000-DSPE摩尔比20∶1,TF质量分数0.10%;所得脂质体TET包封率为97.80%,VCR包封率为93.00%,综合评分为94.44(n=3),与其预测值93.81接近。结论:优化所得处方稳定,可用于制备TF修饰TET与VCR脂质体。     

星点设计-效应面法优化紫杉醇-PluronicP123共聚物胶束处方工艺

目的以星点设计-效应面法优化紫杉醇-Pluronic P123共聚物胶束处方工艺,以期提高难溶性药物紫杉醇在Pluronic P123载体中的包封率和载药量,并对其理化性质进行表征。方法采用薄膜分散法制备紫杉醇-Pluronic P123胶束,以包封率、载药量和胶束溶液药物浓度为考察指标,紫杉醇投药量和水相用量为自变量,采用星点设计-效应面法优化处方,并对模型进行验证,对粒径、体外释放等理化性质进行表征。结果二次多项式非线性回归模型是描述因素与指标关系的最佳模型,根据所优化处方,制得共聚物胶束的包封率约为85.64%,载药量约为1.68%,平均粒径约为(25.2±2.9)nm,zeta电位为(-11.23±2.64)m V。胶束制剂与普通制剂(Taxol注射液)在4 h内的累积释放分别为50.1%和90.5%,前者具有较强的缓释作用。结论通过采用星点设计-效应面优化法确定的处方工艺,可制备具有较高包封率的紫杉醇-Pluronic P123共聚物胶束,所建模型有较好的实用性和预测性。Pluronic P123可有效增溶难溶性药物紫杉醇,并形成具有较强缓释作用的纳米级共聚物胶束制剂,具有应用于肿瘤治疗的潜力。     

一种新型重活化剂的合成及其纳米粒的制备工艺

目的合成一种新型重活化剂酰胺磷定(HI-6)类似物,并制备该化合物纳米粒制剂。方法用两步反应合成目标产物。采用界面聚合法制备纳米粒,以主药投药量、载体材料氰基丙烯酸正丁酯(α-BCA)和Pluronic P85用量为考察因素。以包封率、载药量、平均粒径及归一化值(OD)为考察指标,采用星点设计-效应面优化法优化处方。利用HPLC法测定包封率和载药量,利用激光粒度分析仪测定纳米粒的粒径。结果成功合成产物并制备出纳米粒。通过优化处方制备的样品包封率为63.69%,载药量为6.2%,平均粒径为174 nm。Zeta电位为-7.2 mV。形态为圆形,分布均匀。结论用最优处方制备的载药纳米粒可为后续脑靶向给药奠定基础。     

Tetanus toxoid-loaded transfersomes for topical immunization

Topical immunization is a novel immunization strategy by which antigens and adjuvants are applied topically to intact skin to induce potent antibody and cell-mediated responses. Among various approaches for topical immunization, the vesicular approach is gaining wide attention. Proteineous antigen alone or in combination with conventional bioactive carriers could not penetrate through the intact skin. Hence, specially designed, deformable lipid vesicles called transfersomes were used in this study for the non-invasive delivery of tetanus toxoid (TT). Transfersomes were prepared and characterized for shape, size, entrapment efficiency and deformability index. Fluorescence microscopy was used to investigate the mechanism of vesicle penetration through the skin. The immune stimulating activity of these vesicles was studied by measuring the serum anti-tetanus toxoid IgG titre following topical immunization. The immune response was compared with the same dose of alum adsorbed tetanus toxoid (AATT) given intramuscularly, topically administered plain tetanus toxoid solution, and a physical mixture of tetanus toxoid and transfersomes again given topically. The results indicated that the optimal transfersomal formulation had a soya phosphatidylcholine and sodium deoxycholate ratio of 85:15%, w/w. This formulation showed maximum entrapment efficiency (87.34 +/- 3.81%) and deformability index (121.5 +/- 4.21). An in-vivo study revealed that topically administered tetanus toxoid-loaded transfersomes, after secondary immunization, elicited an immune response (anti-TT-IgG) comparable with that produced by intramuscular AATT. Fluorescence microscopy revealed the penetration of transfersomes through the skin to deliver the antigen to the immunocompetent Langerhans cells.     

右旋糖酐-甲基丙烯酸缩水甘油酯载BMP_2凝胶微球的制备与评价

目的:探讨利用右旋糖酐-甲基丙烯酸缩水甘油酯(DEX-GMA)制备载骨形态发生蛋白2(BMP2)凝胶微球的可行性和制备工艺,并初步探讨其载药、释药特性。方法:以反应温度、DEX-GMA和乳化剂Span-80的加入量和搅拌速率等为考察因素进行正交试验,筛选BMP2-DEX-GMA凝胶微球的最佳制备工艺并对成品性能进行初步检测。结果:经悬浮聚合法可以制备BMP2-DEX-GMA凝胶微球,优选出的制备工艺条件为反应温度30℃、DEX-GMA用量0.6%、Span-80用量0.06%、搅拌速率300r/min,由此制备的BMP2-DEX-GMA凝胶微球形态良好,粒径在20μm～50μm之间,包封率为(78.52±4.34)%,载药量为(10.68±1.34)%,溶胀率为85.9%,稳定性和再分散性良好。结论:该凝胶微球载药系统制备工艺简单,载药量大,可以负载具有生物活性的药物。     

Novel carbopol-based transfersomal gel of 5-fluorouracil for skin cancer treatment: in vitro characterization and in vivo study

Abstract

5-fluorouracil (5-Fu) is an antineoplastic drug, topically used for the treatment of actinic keratosis and nonmelanoma skin cancer. It shows poor percutaneous permeation through the conventionally applicable creams and thus inefficient for the treatment of deep-seated skin cancer. In the present article, transfersomal gel containing 5-Fu was investigated for the treatment of skin cancer. Different formulation of tranfersomes was prepared using Tween-80 and Span-80 as edge activators. The vesicles were characterized for particle size, shape, entrapment efficiency, deformability and in vitro skin permeation. Optimized formulation was incorporated into 1% carbopol 940 gel and evaluated for efficacy in the treatment of skin cancer. 5-Fu-loaded transfersomes (TT-2) has the size of 266.9?±?2.04?nm with 69.2?±?0.98% entrapment efficiency and highest deformability index of 27.8?±?1.08. Formulation TT-2 showed maximum skin deposition (81.3%) and comparable transdermal flux of 21.46?µg/cm²?h. The TT-2-loaded gel showed better skin penetration and skin deposition of the drug than the marketed formulation. Composition of the transfersomal gel has been proved nonirritant to the skin. We concluded that the developed 5-Fu-loaded transfersomal gel improves the skin absorption of 5-Fu and provide a better treatment for skin cancer.     

叶酸偶联壳聚糖载多西他赛纳米粒的制备

目的:探索靶向叶酸受体的多西他赛(DTX)纳米粒的制备方法。方法:利用叶酸活性酯与壳聚糖分子上的氨基反应,制得叶酸偶联壳聚糖(FA-CTS);再通过离子交联法,将DTX作为模型药物,制备叶酸偶联壳聚糖载DTX(FA-CTS/DTX)纳米粒。以载药量、包封率、粒径和跨距为指标,采用星点设计-效应面法优化搅拌速率、DTX加入量、壳聚糖-三聚磷酸钠(CTS-STPP)的质量比,并进行验证。利用激光粒度分析仪测定纳米粒粒径大小及分布,在磷酸盐缓冲液中对载药纳米粒进行体外释药试验。结果:最优处方(处方量为2.5 mg)为搅拌速率为1 300 r/min、DTX加入量为0.58μg,CTS-STPP的质量比为5.55。所制备的FA-CTS/DTX纳米粒平均粒径为(232.8±0.43)nm、包封率为(86.74±0.60)%、载药量为(25.29±3.21)%、跨距为0.039±1.02;30 min内累积释药40.22%,随后缓慢释放,24 h内累积释药80.25%。结论:成功制备具有缓释作用的FA-CTS/DTX纳米粒。     

中心组合设计法优化3′,5′-二辛酰基-氟苷药质体制备工艺

目的采用中心组合设计法优化3′,5′-二辛酰基-5-氟脲嘧啶脱氧核苷药质体的制备工艺.方法以薄膜-超声分散法制备药质体,以平均粒径、包封率、载药量和综合指标为因变量,对药物与磷脂的比例、F-68浓度和三硬脂酸甘油酯浓度3个自变量的各个水平进行二项式拟合,效应面法选取较佳工艺条件并进行预测分析.结果平均粒径、包封率、载药量和综合指标拟合所得多元二次方程的复相关系数分别为0.9393,0.7739,0.9801和0.8369.优化条件制备的药质体的平均粒径、包封率和载药量分别为76nm,97.49%和29.37%.结论中心组合设计有应用简便、预测性好等优点,制备的药质体符合设计要求.     

Preparation and quality assessment of itraconazole transfersomes

Drug-loading transfersomes were prepared with itraconazole, a lipophilic drug, as a model drug to investigate the key factor affecting transfersomes quality and to evaluate their qualities. Drug-loading transfersomes were prepared using film dispersion method. The quality of transfersomes was evaluated by HPLC, transmission electron microscope, particle size analyzer and in vitro release. Itraconazole transfersomes was transparent solution in ivory white color with a mean entrapment efficiency of about 80%. The shape of hollow vesicles was spheroidal with the diameter of approximately 100nm, and the zeta potential of 45mV, which had a good transdermal effect. It can be concluded via single-factor investigation that the quality of transfersomes is significantly affected by solvent, salt ion concentration and homogenization pressure and so on. The preparation method obtained through screening and optimizing formulation and technology is feasible and the quality can be controlled.     

西罗莫司纳米结构脂质载体分散液的制备及其体外释放度考察

目的优化西罗莫司纳米结构脂质载体(sirolimus nanostructured lipid carriers,SRL-NLC)分散液的处方,并考察其体外释放度。方法采用星点设计-效应面法(central composite design-response surface methodology,CCD-RSM)优化SRL-NLC分散液的处方,并以粒径、分布系数、载药量和包封率作为评价指标,采用正相透析袋法考察SRL-NLC分散液在0.4%SDS(十二烷基硫酸钠)溶液中的释放度。结果经优化的SRL-NLC分散液平均粒径82.54 nm、分布系数0.207,Zeta电位-18.0mv、载药量1.829%和包封率91.3%,SRL-NLC分散液在0.4%SDS溶液中能持续释放120 h,累积释放量为60.1%。结论经优化的处方可行性和重现性均较好,SRL-NLC分散液在0.4%SDS溶液中累积释放度为60.1%。     

TRANSFERSOMES FOR VACCINE DELIVERY: A POTENTIAL APPROACH FOR TOPICAL IMMUNIZATION

Delivery of vaccine is a major impediment to ensuring vaccine efficacy and compliance. Existing vaccine delivery approaches have their limitations and these led to the development of novel approaches for vaccine delivery. Non-invasive vaccine delivery is in demand and for this skin appears to be a potential sites that elicit immune responses. Topical application of antigen and adjuvant directly on the skin is termed as topical vaccination. It has potential to reduce physical skin penetration by injection and provide potentially effective vaccines. In the present study novel carrier, transfersomes were used for topical delivery of tetanus toxoid (TT). Transfersomes were prepared by reverse phase evaporation method and they were evaluated for shape, size, entrapment efficiency and deformability index. Transfersomal formulation with optimal concentration of Soya phosphatidylcholine (SPC) and sodium deoxycholate (85:15 w/w) showed entrapment efficiency of 39.8±0.032 and deformability index of 16.4. In-vivo study revealed that topically given TT containing transfersomes after boosting produced immune responses (0.877±0.081) that is as good as that produced by subcutaneously given alum adsorbed TT (0.948±0.088). Furthermore, fluorescence microscopy has confirmed the bioactive delivery through the skin layers. A brief comparative study has been done with liposomes.     

毛蕊花苷固体脂质纳米粒的处方优选及其质量评价

目的:优选毛蕊花苷(VER)固体脂质纳米粒(SLN)的处方,并对VER-SLN质量进行评价。方法:采用乳化超声分散法制备VER-SLN,以包封率为评价指标,以药脂质量比、单硬脂酸甘油脂用量、泊洛沙姆188用量、豆磷脂用量为考察因素,通过正交试验对处方进行优化,同时以载药量、粒径、Zeta电位、包封率、稳定性及体外累积释放度为指标评价其质量。结果:最佳制备处方为药脂质量比为1∶75,单硬脂酸甘油脂的用量为0.6 g,泊洛沙姆188用量为0.5 g,豆磷脂用量为0.2 g。所制得的VER-SLN外观形态圆整,粒度分布均匀,平均粒径为(109±17)nm,Zeta电位为(-23±0.91)mV,平均包封率为96.66%,平均载药量为2.27%。体外释放结果表明,VER原料药体外8 h累积释放完全,VER-SLN体外4 h累积释放率为47.2%,48 h可达到92.9%。结论:该制剂处方设计合理,制备工艺稳定,乳化超声分散法制备的VER-SLN质量符合要求,可达到使药物缓慢释放的效果。     

2,5-二酮-3,6-二(N-富马酰基-4-氨基丁基)哌嗪(FDKP)载胰岛素微球的制备及表征

目的：制备并表征载胰岛素的2,5-二酮-3,6-二（N-富马酰基-4-氨基丁基）哌嗪（FDKP）微球。方法以 FDKP 作为载体,制备载胰岛素微球（FDKP-INS）,利用 Box-Behnken 试验设计对载药微球的载药量及粒径进行筛选。使用扫描电子显微镜（SEM）对微球形态进行观察,同时以差示扫描量热法（DSC）考察胰岛素与 FDKP 间相互作用。结果最优处方工艺条件为：混悬液 pH 值为4.56,投药比为15.97%（W/ W）,搅拌时间为2.4 h,在此工艺条件下制备的微球载药量为13.23%,粒径为4.515μm,与预测值接近;显示扫描图谱表明,胰岛素与 FDKP 间存在相互作用;扫描电子显微镜显示空白 FDKP 微球外形呈玫瑰花球形状,形态较完整,且表面呈现多孔隙,载药微球表面较平滑,胰岛素填充入空白微球孔隙中。结论经过优化的处方工艺所制得的载胰岛素 FDKP 微球具有良好的外观性状和较高的载药量。