唑来膦酸脂质体的制备及对肿瘤细胞的抑制作用

目的制备唑来膦酸脂质体并探讨其对多发性骨髓瘤RPMI 8226肿瘤细胞的抑制作用。方法采用薄膜分散法制备唑来膦酸脂质体,采用磺酰罗丹明B(SRB)染色法考察唑来膦酸脂质体对肿瘤细胞的体外抑制作用。结果制备所得唑来膦酸脂质体的平均粒径为179.9 nm,包封率40.22%,载药量3.54%,平均电位-24.3 m V。唑来膦酸针剂与唑来膦酸脂质体对肿瘤细胞均有抑制增殖作用。结论采用薄膜分散法制得的唑来膦酸脂质体具有较好的稳定性,粒径分布均匀,符合制剂学要求。采用脂质体包裹后的唑来膦酸对肿瘤细胞的抑制作用明显强于唑来膦酸针剂。     

唑来膦酸的合成

目的　以 2 - (咪唑 - 1-基 )乙酸盐酸盐为原料合成唑来膦酸。方法　采用反应物浓磷酸自身作为溶剂 ,以无水乙醇作为结晶溶剂 ,合成唑来膦酸。结果和结论　避免了有毒有机溶剂的使用 ,将产物收率由 4 1.0 %提高到了 5 8.8%。唑来膦酸的结构经1HNMR、MS得到确证。     

唑来膦酸的合成

咪唑和氯乙酸乙酯通过非均相反应合成咪唑乙酸乙酯，酸性水解得到咪唑乙酸盐酸盐，和三氯化磷、磷酸反应后再水解制得唑来膦酸。原料价廉易得，操作简便，反应总收率36％。     

唑来膦酸的合成

目的：改进唑来膦酸的合成工艺。方法：采用新的KF／Al2O3催化体系和PEG400溶剂系统，进行关键中间体和目标物的合成研究，结果：合成关键中间体的反应时间由原法的18h缩短为3h，收率也由66％提高到77．5％，目标物的收率也由原法的41％提高到50．7％。结论：新方法收率提高，操作简便，合成周期也缩短，适合于工业生产。     

唑来膦酸致全身皮疹

1例57岁女性患者因重度骨质疏松症给予碳酸钙和骨化三醇口服,1周后,给予唑来膦酸5 mg溶于0.9%氯化钠注射液100 ml静脉滴注。第2天,患者出现发热、全身皮疹,头晕、乏力、关节疼痛及双下肢肌肉酸痛。立即给予地塞米松、氯雷他定。第7天,患者头晕、乏力、关节疼痛及双下肢肌肉酸痛等症状消失,体温降至正常。第10天皮疹完全消退。     

唑来膦酸急性毒性试验研究

按新药审批“急性毒性试验”的要求与方法 ,用昆明小鼠以灌胃、静脉注射两种给药途径进行了唑来膦酸急性毒性试验。结果显示 ,昆明小鼠口服给药受试物 LD50 为 713 .5 ( 667.8～ 763 .2 ) mg/kg;静脉给药 LD50 为 13 .8± 0 .8( 12 .6～ 14 .3 ) mg/kg。     

唑来膦酸的临床研究进展

目的:综述第3代双膦酸类药物--唑来膦酸的临床研究进展.方法:通过查阅文献,总结唑来膦酸在药动学、安全性和临床应用方面的研究进展.结果:唑来膦酸可以抑制破骨细胞调节的骨吸收,降低血清钙水平,能有效地用于恶性高钙血症的治疗.结论:唑来膦酸是高效的破骨细胞骨吸收抑制剂,它不仅能用于恶性高钙血症的治疗,并有望成为治疗骨质疏松症等骨骼疾病的新一类高效药物.     

唑来膦酸的药效学研究进展

目的:综述第3代双膦酸类药物-唑来膦酸的药效学研究进展。方法:通过查阅文献总结唑来膦酸在动物和人体试验中的 药效学。结果:唑来膦酸可以抑制破骨细胞调节的骨吸收,降低血清钙水平,能有效地用于恶性高钙血症的治疗。结论:唑来膦酸是 最高效的破骨细胞骨吸收抑制剂,它能有效地治疗恶性高钙血症,并有望成为治疗骨质疏松症的新一类高效药物。     

唑来膦酸的临床应用

目的为临床应用唑来膦酸提供参考。方法综述唑来膦酸的药理作用及临床应用。结果唑来膦酸临床用于治疗恶性高钙血症,控制恶性肿瘤骨转移,并能改善多发性骨髓瘤,前列腺癌、乳腺癌、肺癌等恶性肿瘤治疗的疗效。结论唑来膦酸可提高肿瘤患者的生活质量和延长患者的生存期。     

冰片修饰的姜黄素阳离子脂质体的制备及其脑靶向作用研究

目的 制备冰片修饰的姜黄素阳离子脂质体（curcumin-loaded modifying borneol cationic liposomes,Cur-BCLPs）,鼻腔给药后考察其在大鼠体内的药动学行为并对其脑组织分布进行研究。方法 采用乙醇注入法制备Cur-BCLPs;透射电镜观察阳离子脂质体的形态;激光粒度仪考察粒径;超速离心法测定其包封率及载药量;以姜黄素混悬液（curcumin suspension,Cur-Sol）和冰片-姜黄素混悬液（borneol curcumin suspension,BO-Cur-Sol）为对照组,考察大鼠鼻腔给药Cur-BCLPs的体内药动学过程,并测定其在大鼠脑组织的浓度,运用DAS 2.0软件拟合药动学参数。结果 阳离子脂质体外观呈圆形或类圆形,平均粒径为（105.99±2.40）nm,包封率和载药量分别为（81.95±1.03）%和（4.28±0.46）%;体内药动学结果显示,Cur-Sol、BO-Cur-Sol和Cur-BCLPs的半衰期（T_1/2）分别为（4.27±1.53）h,（3.98±0.24）h和（6.01±0.63）h,AUC_0→t分别为（224.38±21.95）μg·h·L^-1,（243.40±12.26）μg·h·L^-1和（562.28±24.30）μg·h·L^-1,清除率分别为（1.82±0.36）L·h^-1·kg^-1,（1.72±0.11）L·h^-1·kg^-1和（0.78±0.03）L·h^-1·kg^-1,滞留时间分别为（4.28±0.23）h,（4.41±0.15）h和（8.09±0.17）h。脑组织分布结果显示,Cur-Sol、BO-Cur-Sol和Cur-BCLPs的AUC_0→t分别为（29.82±1.10）μg·h·g^-1,（35.47±1.75）μg·h·g^-1和（54.06±3.90）μg·h·g^-1,清除率分别为（15.73±0.84）L·h^-1·kg^-1,（13.23±0.52）L·h^-1·kg^-1和（8.52±0.92）L·h^-1·kg^-1。结论 Cur-BCLPs经鼻腔给药后显著提高姜黄素体内和脑组织蓄积量并且延缓消除。     

槐定碱阳离子脂质体的制备及体外抗肿瘤活性的研究

目的制备槐定碱阳离子脂质体,并探讨其对肿瘤细胞的抑制作用。方法采用主动载药法制备槐定碱阳离子脂质体,并对其进行表征研究,采用MTS方法考察槐定碱阳离子脂质体对3种肿瘤细胞的抑制作用。结果制备得到的槐定碱阳离子脂质体呈类圆形,表面光滑,其平均粒径和聚分散指数分别为242.2 nm和0.180,表面电荷为+32.5 m V,其包封率和载药量分别为88.62%和5.97%。槐定碱阳离子脂质体对3种肿瘤细胞的IC50值均明显高于槐定碱,而空白阳离子脂质体对细胞并无明显的抑制作用。结论采用阳离子脂质体作为槐定碱的载体有利于将药物透过细胞膜,提高抗肿瘤作用,值得进行深入的系统研究。     

唑来膦酸及其制剂的微生物学检查方法研究

目的：建立唑来膦酸的微生物限度检查方法以及唑来膦酸注射液的无菌检查方法。方法：按《中国药典》2015年版四部要求进行方法适用性试验。结果：唑来膦酸微生物限度检查法采用薄膜过滤法,需氧菌总数、霉菌和酵母菌总数计数方法适用性试验5种试验菌回收率比值均在0.5～2之间;控制菌大肠埃希菌检查试验组可检出大肠埃希菌。唑来膦酸注射液无菌检查方法采用薄膜过滤法,6种试验菌均能检出。结论：唑来膦酸及唑来膦酸注射液的《中国药典》2015年版的微生物学检查方法均采用薄膜过滤法。     

Acacia-Gelatin Microencapsulated Liposomes: Preparation,Stability, and Release of Acetylsalicylic Acid

Liposomes of dipalmitoylphosphatidylcholine (DPPC) containing acetylsalicylic acid (ASA) have been microencapsulated by acacia-gelatin using the complex coacervation technique as a potential oral drug delivery system. The encapsulation efficiency of ASA was unaltered by the microencapsulation process. The stability of the microencapsulated liposomes in sodium cholate solutions at pH 5.6 was much greater than the corresponding liposomes. The optimum composition and conditions for stability and ASA release were 3.0% acacia-gelatin and a 1- to 2-hr formaldehyde hardening time. Approximately 25% ASA was released in the first 6 hr from microencapsulated liposomes at 23°C and the kinetics followed matrix-controlled release (Q t ^l/2). At 37°C, this increased to 75% released in 30 min followed by a slow constant release, likely due to lowering of the phase transition temperature of DPPC by the acacia-gelatin to near 37°C. At both temperatures, the release from control liposomes was even more rapid. Hardening times of 4 hr and an acacia-gelatin concentration of 5% resulted in a lower stability of liposomes and a faster release of ASA. It is concluded that under appropriate conditions the microencapsulation of liposomes by acacia-gelatin may increase their potential as an oral drug delivery system.     

Oxygen Radical-Mediated Pulmonary Toxicity Induced by Some Cationic Liposomes

Purpose. The objectives of this study are to investigate the toxicityassociated with polycationic liposomes and to elucidate the underlyingmechanism. We tested the hypothesis that the positive charge of liposomesis a key determinant of toxicity by testing differently chargedliposomes in mice. Methods. Differently charged liposomal systems including cationicliposomes, LipofectAMINE and DOTAP, and neutral and negativeliposomes were evaluated for their toxicity after pulmonaryadministration in mice. LDH assay and differential cell counts were performedto measure toxicity and pulmonary inflammation, respectively. Reactiveoxygen intermediates (ROI) were assessed by chemiluminescence. Results. Instillation of cationic liposomes eliciteddose-dependent toxicity and pulmonary inflammation. This effect was more pronouncedwith the multivalent cationic liposome LipofectAMINE as comparedto the monovalent cationic DOTAP. Neutral and negative liposomes didnot exhibit lung toxicity. Toxicity associated with cationic liposomescorrelated with the oxidative burst induced by the liposomes.LipofectAMINE induced a dose-dependent increase in ROI generation. Thiseffect was less pronounced with DOTAP and absent with neutral andnegative liposomes. Conclusions. ROI play a key role in cationic lipid-mediated toxicity.Polyvalent cationic liposomes cause a release of ROI which areresponsible for the pulmonary toxicity.     

Comparison of Cell Proliferation and Toxicity Assays Using Two Cationic Liposomes

The present study compares different cytotoxicity and cell proliferation assays including cell morphology, mitochondrial activity, DNA synthesis, and cell viability and toxicity assays. CaSki cells were exposed to two cationic liposomal preparations containing dimethyldioctadecyl-ammonium bromide (DDAB), dioleoylphosphatidylethanolamine (DOPE) and a commercial transfection-reagent DOTAP(N[l-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium-methylsulfate). The results provided by these assays were similar. However, the lactate dehydrogenase assay was more sensitive in measuring early damages of cell membranes than the Trypan blue assay. Also, cell morphology showed early toxic changes, such as cytoplasmic vacuolization and cell shrinking, and it should be included with such toxicity evaluations. DDAB:DOPE was more toxic than DOTAP. The cells treated with DOTAP at 10 µM were surviving as well as the control cells, while DOTAP at 40 µM and DDAB: DOPE at 10 µM had slight toxic effects on CaSki cells. The most toxic effects were seen in CaSki cells after treatment with DDAB: DOPE at 40 µM.     

盐酸5-氨基酮戊酸脂质体的制备与表征

目的 研究盐酸5-氨基酮戊酸(5-ALA)脂质体处方组成及制备工艺. 方法 采用薄膜分散 pH梯度法制备5-ALA脂质体,以包封率为评价指标进行正交实验筛选出最佳处方,采用高效液相色谱(HPLC)法测定其包封率. 并对其粒径、电位等理化性质进行研究. 结果最佳处方工艺为：卵磷脂与胆固醇的质量比为6：1,卵磷脂与5-ALA质量比为6：1,孵育温度为60 ℃. 所制脂质体为乳白色,平均粒径为100 nm,Zeta电位为-40 mV,平均包封率为65.0%. 结论 该制备方法 得到的5-ALA脂质体处方合理,工艺可行,包封率较高.     

第3代双膦酸盐类药物--唑来膦酸

介绍唑来膦酸在药效学、药代动力学和临床应用等方面的研究进展.唑来膦酸可抑制破骨细胞调节的骨吸收,降低血清钙水平,抑制肿瘤细胞的生长,能有效治疗恶性肿瘤所致高钙血症、晚期肿瘤骨转移和变形性骨炎,还有望用于骨质疏松症的治疗.     

顶空气相色谱法测定唑来膦酸中有机溶剂残留量

目的采用顶空气相色谱法测唑来膦酸中有机溶剂残留量。方法用DB-624毛细管柱（30m×0．32mm,1．8μm）,柱温为起始温度40℃,维持10min,再以每分钟35℃的速率升温至220℃,维持5min;进样口温度为250℃;氢火焰离子化检测器（FID）;检测器温度为280℃。结果6种溶剂的分离度良好,在所考察的质量浓度范围内线性关系良好,r=0．9999,平均回收率为96．9％-102．3％,检出限为7．86pg／mL-75．0ng／mL。结论所用方法简单、灵敏度高,可用于唑来膦酸中有机溶剂残留量的限度检查。