Box-Behnken效应面法优化制备主动载药盐酸小檗碱脂质体

目的采用Box-Behnken效应面法筛选最佳处方,制备盐酸小檗碱脂质体。方法采用薄膜分散-p H梯度法制备脂质体,分别以磷脂与胆固醇质量比、脂药质量比、外水相p H值、孵化温度为考察对象,以包封率、粒径和载药量为评价指标,采用4因素3水平Box-Behnken效应面设计法筛选盐酸小檗碱脂质体的最佳处方。采用阳离子交换树脂微柱离心法测定包封率,动态激光散射法测定脂质体的粒径,并采用透射电镜观察制得的脂质体形态。结果最优处方工艺条件为磷脂与胆固醇质量比为3.38∶1,脂药质量比为22∶1,外水相p H为6.88,孵化温度为59℃。以最优处方制备的盐酸小檗碱脂质体平均粒径、包封率、载药量与预测值偏差较小。结论采用Box-Behnken效应面法优化盐酸小檗碱脂质体工艺处方是可行的。     

Box-Behnken效应面法优化制备氯诺昔康柔性脂质体

目的采用Box-Behnken效应面法筛选最佳处方,制备氯诺昔康柔性脂质体。方法采用薄膜分散水化法制备脂质体,分别以磷脂浓度、磷脂与胆固醇质量比、脂药质量比、吐温80与总脂质质量比为考察对象,以包封率、粒径为评价指标,采用4因素3水平Box-Behnken效应面设计法筛选氯诺昔康柔性脂质体的最佳处方。采用葡聚糖G50微柱离心法测定包封率,动态激光散射法测定脂质体的粒径和ζ-电位,采用透射电镜观察制得的脂质体形态,并考察脂质体的体外释放。结果最优处方工艺条件为磷脂质量浓度为16.94 g·L~(-1),磷脂与胆固醇质量比为4.46∶1.00,总脂质与吐温80质量比8.12∶1.00,脂药质量比18.65∶1.00。以最优处方制备的氯诺昔康柔性脂质体平均粒径较小(93.86±7.58)nm、ζ-电位较好[-(20.21±2.31)mV]、包封率较高(90.23±1.46)%,实际值与预测值偏差较小。质量分数为82.06%的药物在24h内从脂质体中释放出来,具备明显的缓释特性。结论采用Box-Behnken效应面法优化氯诺昔康柔性脂质体工艺处方是可行的。     

棓丙酯脂质体的制备与表征

目的制备棓丙酯脂质体,并对其进行理化性质的表征和释放度的评价。方法采用薄膜分散法制备棓丙酯脂质体,超滤离心法测定脂质体的包封率,正交设计优化处方,并对其包封率、粒径、Zeta电位、形态及体外释放行为进行综合评价。结果正交设计优化最终处方为磷脂浓度5 mg.mL-1、药脂比1∶5、磷脂胆固醇比5∶1、水化介质离子强度20 mmol.mL-1,所得脂质体包封率为89.6%、粒径为181.3 nm、Zeta电位为-21.8 mV、4 h体外释放达到80%。结论制备的棓丙酯脂质体包封率高,粒径小而均一,体外释放完全。     

苯丁酸氮芥脂质体的制备及处方因素考察

目的 制备苯丁酸氮芥脂质体并优化其处方。方法 薄膜超声分散法制备苯丁酸氮芥脂质体,采用微柱离心-HPLC法测定其包封率,以包封率为考察指标,研究膜材比、药脂比、水相介质pH值以及磷脂浓度等因素对脂质体包封率的影响;通过正交试验对处方进行优化,并进行质量评价。结果 苯丁酸氮芥脂质体优化后的制备处方为胆固醇与磷脂质量比1∶3、药脂比1∶10、水相介质pH值为7.4、磷脂浓度为0.3%。按该处方制得的苯丁酸氮芥脂质体包封率>87%,平均粒径为84.71 nm,PDI为0.167。结论 优选处方稳定可行,制备的苯丁酸氮芥脂质体包封率高、粒径小且均匀。     

维A酸脂质体的制备工艺研究

目的:提供一种制备稳定维A酸脂质体的新方法。方法:采用乙醇注入法制备维A酸脂质体,通过单因素试验和四因素三水平正交试验,以包封率为主要指标,优选药脂比、磷脂与胆固醇之比、水合介质(0.01 mol/L磷酸盐缓冲液)的pH和用量;并制备样品进行稳定性考察。结果:以药脂比为1∶10(维A酸用量1.0 mg),磷脂与胆固醇之比为4∶1,水合介质的pH为6.5、用量为30ml为最佳工艺;所制维A酸脂质体的包封率约为80%,平均粒径约为150 nm。在室温条件下密封放置10 d及4℃下保存6个月,其平均粒径及包封率差异无统计学意义(P>0.05)。结论:该制剂制备工艺简单可行;制剂处方合理,包封率较高,且在短期内稳定性良好。     

复乳法制备龙胆苦苷脂质体

目的以大豆卵磷脂和胆固醇等为载体制备龙胆苦苷脂质体。方法采用复乳法考察乳化剂的用量、药脂比、磷脂与胆固醇质量比等因素对脂质体制备过程的影响;以外观形态和包封率为考察指标,优化龙胆苦苷脂质体的最佳制备工艺。结果复乳法制备的龙胆苦苷脂质体最佳工艺为：大豆卵磷脂与胆固醇质量比为6∶1,药脂比为1∶8,助乳剂量为30∶1,包封率可达82%。结论复乳法制备龙胆苦苷多囊脂质体均匀、稳定,粒径分布（414.8±50.0）nm,Zeta电位为（29.2±5.0）mV。     

尼莫地平脂质体的制备与理化性质研究

［摘要］目的制备尼莫地平脂质体并考察其理化性质。方法采用单因素考察及正交优化实验,对尼莫地平脂质体的处方组成进行了筛选,对制备工艺进行了优化,同时对尼莫地平脂质体包封率、粒子粒径、磷脂氧化率等进行了测定。结果尼莫地平脂质体最佳处方中磷脂与胆固醇之比为1：4,二硬脂酰磷脂酰甘油与磷脂和胆固醇混合物之比为1：10,主药与总脂质量之比为1：40。尼莫地平脂质体平均粒径为145.2 nm,包封率为97.6%。结论制备的尼莫地平脂质体包封率较高,稳定性良好,符合临床用药要求。     

盐酸青藤碱柔性纳米脂质体的制备及质量考察

目的制备盐酸青藤碱柔性纳米脂质体,并研究其质量。方法用微柱离心法分离脂质体和游离药物,用HPLC法测定包封率。考察薄膜分散法、逆向蒸发法和pH梯度法对包封率的影响,并采用正交实验设计筛选盐酸青藤碱柔性纳米脂质体的最佳处方,测定其粒径。结果三种制备方法得到的药物脂质体的包封率大小依次为:pH梯度法>逆向法>薄膜法。正交实验设计中影响包封率的最显著因素为磷脂与胆酸钠的质量比,其次为外水相pH值,药脂比与孵育温度影响较小。最佳处方平均粒径128.4 nm,4℃放置30 d未产生聚积或沉淀。结论pH梯度法适合于制备盐酸青藤碱柔性纳米脂质体,工艺可行,产品质量稳定。     

灯盏花素脂质体的制备工艺

目的研究灯盏花素脂质体的制备工艺。方法采用薄膜蒸发-探头超声法和冷冻干燥法制备灯盏花素脂质体,在单因素考察基础上采用正交试验设计。以包封率为评价指标,筛选脂质体制备的最佳工艺条件。冻干品水合后,在电镜下观察灯盏花素脂质体的形态,利用马尔文测定仪测定脂质体的粒径,用RP-HPLC法测定其包封率。结果灯盏花素脂质体的最佳工艺处方为药脂比1∶5,SPC∶CH为2∶1,二氯甲烷用量为10 ml。冻干保护剂蔗糖用量为10%。制备3批脂质体,包封率平均为87.5%,平均粒径为378.3 nm。结论所制脂质体包封率较高,粒径分布较均匀。     

颌面血管瘤局部注射用平阳霉素脂质体的制备和优化

目的制备平阳霉素脂质体,建立包封率测定方法,考察处方影响因素,优化处方。方法采用硫酸铵梯度法制备平阳霉素脂质体;以Sephadex G-50微柱离心法分离脂质体和游离药物,应用HPLC法测定药物含量,计算包封率;通过单因素考察优化硫酸铵梯度法的处方工艺,考察不同处方因素对包封率的影响。结果 Sephadex G-50微柱可完全吸附平阳霉素游离药物,空白脂质体回收率为98.3%~101.2%;平阳霉素浓度在5~200μg·m L-1内与峰面积线性关系良好(r=0.9999),日内和日间精密度(RSD)均<2%,回收率为98.3%~100.1%。通过单因素考察优化平阳霉素脂质体的处方为:磷脂100 mg;胆固醇20 mg;平阳霉素15 mg。优化工艺为采用浓度为250 mmol·L-1硫酸铵溶液制备空白脂质体,加入平阳霉素溶液后在40℃水浴中载药40 min。结论硫酸铵梯度法适用于制备平阳霉素脂质体,微柱离心法测定包封率准确可靠,处方工艺优化后的平阳霉素脂质体的包封率可以达到63.7%。     

微柱离心-药脂比测定脂质体药物包封率

目的建立一种准确测定脂质体包封率的方法。方法以氟吡洛芬、酮洛芬、拓扑替康为模型药物脂质体,通过葡聚糖凝胶(Sephadex G-50)微柱离心去除游离药物,以HPLC法测定药物含量,以定磷法测定磷脂含量,通过药脂比测定药物包封率。结果无需将脂质体完全洗脱,通过检测洗脱脂质体中的药脂比能够准确测定脂质体包封率;对于水溶性药物拓扑替康,若将脂质体完全洗脱,则可能无法将游离药物完全分离。结论在一定条件下微柱离心去除游离药物,通过药脂比测定包封率方便准确。     

氟比洛芬脂质体的制备和体外透皮实验

目的研究氟比洛芬脂质体的制备和体外透皮扩散。方法乙醇注入法制备不同粒径的氟比洛芬脂质体,微型柱离心法分离脂质体和游离药物,采用Franz扩散池,考察氟比洛芬脂质体透过单位面积鼠皮的累积量、渗透系数和皮内滞留量。结果相对于大粒径脂质体,小粒径脂质体显著提高氟比洛芬的透皮速率;开放式给药组的累积透过量、渗透系数和皮内滞留量高于封闭组。结论脂质体给药方式和脂质体的粒径是影响氟比洛芬体外透皮的主要因素。     

微柱离心结合双波长考马斯亮蓝法/酶标仪法测定牛血清白蛋白脂质体的包封率

目的:测定牛血清白蛋白(BSA)脂质体的包封率。方法:采用微柱离心结合双波长考马斯亮蓝法/酶标仪法测定,即先用葡聚糖凝胶DEAE-Sephadex A50微柱离心法将游离药物与脂质体进行分离,再筛选样品与染料体积比及反应时间,确定了双波长(595、450nm)考马斯亮蓝法/酶标仪法的测定条件并进行了方法学考察;考察了微柱对空白脂质体的回收率和微柱的分离效率;对BSA脂质体的包封率进行了测定。结果:确定的测定条件为样品与染料体积比为1∶3、反应时间10min,该法平均回收率为99.56%,RSD小于2.73%;微柱对空白脂质体的平均回收率为97.78%,分离效率大于90%;BSA脂质体的平均包封率为32.91%(n=3)。结论:本法实现了游离蛋白质与脂质体快速有效的分离,方法简单易行,与普通凝胶层析柱分离法相比,本法对样品的稀释倍率大大降低,适用于测定痕量大分子蛋白质脂质体的包封率。     

Kit formulation for the preparation of radioactive blue liposomes for sentinel node lymphoscintigraphy

A radioactively labeled blue liposome formulation was developed for use in presurgical lymphoscintigraphy and intraoperative sentinel node localization to avoid the differing injection site clearance kinetics of the conjunctive use of separate formulations of low molecular weight blue dye and radioactively labeled macromolecular sulfur colloid. Blue liposomes containing glutathione in the internal aqueous phase were prepared from blue dyed lipids obtained by covalently binding Reactive Blue II to phosphatidylethanolamine (PE) fractions of phospholipid extracts. Four phospholipid extracts with differing PE fractions and a centrifugation technique were evaluated with the goal of maximizing the blue color intensity of the formulation. Stability of the formulations was evaluated by studying radiolabeling efficiency (using a membrane permeable lipophilic carrier of the commonly used diagnostic radionuclide, technetium-99m) and particle-size distribution over 30-60-day periods. Blue color was not altered by varying the PE content, while centrifugation was an effective and convenient method to maximize the blue color intensity of the final preparation. The particle size distribution of the prepared liposomes ranged from 200-300 nm (considered ideal for lymphoscintigraphy studies) and did not change significantly, while radiolabeling efficiency exceeded 80% for up to 1 month. The described kit formulation for the preparation of radiolabeled blue liposomes is suitable for commercial production allowing widespread clinical use. The combination of a means of visual identification and tracking of the liposomes through the lymphatic channels along with the ability to trace the preparation using standard radiation detection instrumentation provides the surgeon with an improved radiolabeled compound for lymphoscintigraphy and intraoperative sentinel lymph node identification.     

硫酸阿米卡星多囊脂质体的制备及评价

目的 制备硫酸阿米卡星多囊脂质体(amikacin sulfate multivesicular liposomes, AMK-MVLs),对其进行质量评 价,并考察了其体外抗菌活性。方法 采用复乳法制备AMK-MVLs混悬液Ⅰ,Box-Behnken效应面法优化筛选最佳处方,采 用生理盐水洗涤后调整药物浓度得AMK-MVLs混悬液。采用光学显微镜、激光粒度仪、差示扫描量热(differential scanning calorimeter,DSC)考察制剂的理化性质,采用透析法考察其体外释放规律,通过微量稀释法初步考察其体外抗菌活性。结 果 优化得到AMK-MVLs混悬液Ⅰ的最佳处方为：大豆磷脂与胆固醇质量比为1.91:1,三油酸甘油酯用量为1.02%,PVA用量为 0.62%。AMK-MVLs呈堆叠有无数囊泡的非同心球状,AMK-MVLs混悬液包封率(87.12±1.55)%,平均粒径为11.93 μm。DSC 结果表明,AMK以无定型状态存在于脂质体内。体外释放结果显示AMK-MVLs混悬液在72 h时释药约80%。体外溶血实验表 明,AMK-MVLs脂质体粒子浓度低于400 μg/mL时无溶血风险。体外抗菌实验结果显示,相较于AMK溶液,AMK-MVLs混悬液对E. coli、 P. aeruginosa、S. aureus 3种细菌具有更好的抗菌效果。结论 成功制备了一种硫酸阿米卡星多囊脂质体,其粒径分布均匀、包 封率高,释药规律符合Higuchi动力学模型,具有增强的抗菌活性。     

盐酸小檗碱脂质体的制备及其含量、包封率的测定

目的：制备盐酸小檗碱脂质体,并测定其包封率和脂质体各成分含量。方法：采用硫酸铵梯度法制备盐酸小檗碱脂质体,以超速离心法、微柱法、超滤法对盐酸小檗碱脂质体包封率的测定方法进行研究,以HPLC-ELSD测定脂质体各成分含量。结果：超速离心法能将未包封药物与脂质体很好地分离,最佳超速离心条件：离心速度为60000 r·min-1,离心时间为1 h,离心温度为10℃,脂质浓度为6 mg·ml-1。结论：包封率测定方法具有简单、快速分离等优点。 HPLC-ELSD能够同时测定脂质体各成分含量。     

Pilocarpine hydrochloride liposomes: characterization in vitro and preliminary evaluation in vivo in rabbit eye

Liposomes containing either pilocarpine hydrochloride or pilocarpine free base were prepared by the sonication method. This manufacturing process yielded after removal of non-encapsulated solute, small multilamellar vesicles (MLV) as was confirmed by electron microscopy examinations. For an identical liposomal composition, the encapsulation capacity and the drug content of the liposomes were drastically higher for pilocarpine hydrochloride than for pilocarpine free base. Investigation of the preparative parameters revealed that increasing the initial amount of drug decreased the drug content and the encapsulation efficiency of the liposomes formed. Since fixed amounts of lipids were used, the volume sequestration rate decrease was attributed to a moderate viscosity increase of the dispersion medium. Increase of phospholipid concentration at a constant ratio of cholesterol and dicetylphosphate to phosphatidylcholine reduced the aqueous volume entrapped per mg of lipid and subsequently the pilocarpine content in the liposomes. Negatively charged liposomes gave larger rates of pilocarpine hydrochloride and aqueous volume encapsulation than neutral liposomes but, on the contrary, positively charged liposomes gave the lowest rates of pilocarpine hydrochloride and aqueous volume encapsulation. Thus, for drug carrying the same net charge as the phospholipids an increase in the surface charge density of the liposome was not only ineffective, but actually resulted in a lower drug encapsulation due to electrostatic repulsion. Preliminary in vivo results on rabbit eyes suggested that the liposomal vehicle was probably unable to improve sufficiently the corneal penetration of pilocarpine to reach satisfactory therapeutic levels when administered at lower concentrations than commonly used.     

重酒石酸长春瑞滨脂质体包封率测定方法比较

目的制备重酒石酸长春瑞滨脂质体,比较葡聚糖凝胶微柱离心法和阳离子交换树脂离心法测定重酒石酸长春瑞滨脂质体包封率的差异。方法pH梯度法制备重酒石酸长春瑞滨脂质体,分别以葡聚糖凝胶Sephadex G-50和阳离子交换树脂装柱,离心分离脂质体和游离药物,采用HPLC法测定药物含量,计算包封率,并用SPSS软件进行t检验。结果两种方法测定不同载药量的脂质体包封率结果均无显著性差异(P>0.05)。结论葡聚糖凝胶微柱离心法和阳离子交换树脂离心法均可用来测定重酒石酸长春瑞滨脂质体包封率,优选阳离子交换树脂离心法。     

Hydration of lipid films with an aqueous solution of Quil A: a simple method for the preparation of immune-stimulating complexes

Immune-stimulating complexes (ISCOMs) are stable colloidal complexes of the adjuvant Quil A, cholesterol and phospholipid, which are effective carriers for subunit vaccines. The techniques currently available for the preparation of ISCOMs from the constituent components are rather complex and are based on either centrifugation or dialysis. This note reports a new simple procedure for the preparation of ISCOM matrices based on hydration of a cholesterol/phospholipid film with an aqueous solution of Quil A. It is demonstrated that ISCOM matrices do not form in the absence of phospholipid when prepared by this method. Further, the ratio by weight of phospholipid to either cholesterol or Quil A must be greater than that required for preparation by either dialysis or centrifugation. Photon correlation spectroscopy, negative stain transmission electron microscopy and centrifugation through a sucrose gradient demonstrate that ISCOM matrices can be prepared from cholesterol/lipid films by hydration with an aqueous solution of Quil A when the ratio of phospholipid:cholesterol:Quil A by weight is 6:1:4, respectively. Lower ratios of phospholipid:cholesterol reduce the efficiency of ISCOM formation while higher ratios produce systems containing a mixture of ISCOMs together with liposomes.     

In vivo studies on the role of complement in the clearance of liposomes in rats and guinea pigs.

The ability of complement (C) system to remove liposomes from blood circulation was examined in vivo using rat and guinea pig as models. Although the liposomes were not degraded in guinea pig serum in vitro, they were degraded remarkably in guinea pig circulation, as assessed by the urinary excretion of [3H]inulin released from liposomes. The suppression of rat C system to 64% normal C hemolytic activity by treating animals with K76COOH agent resulted in a significant decrease in both the uptake of liposomes by liver and the release of [3H]inulin, providing in vivo evidence for C-mediated clearance of liposomes in rats via uptake by macrophages and degradation in blood circulation, respectively. On the other hand, the K76COOH-induced suppression of C (70% normal hemolytic activity) in guinea pigs slightly increased both the hepatic uptake and the release of [3H]inulin. In addition, the hepatic uptake and in vivo degradation in guinea pigs varied in an opposite manner when the animals were preloaded by empty liposomes or when the liposome size and cholesterol content varied. These results suggest there is a difference between the factors involved in liposome degradation and the factors involved in hepatic uptake and also support the likelihood that there is no C-mediated degradation in guinea pigs.