Lindner Lars H, Hossann Martin, Vogeser Michael, Teichert Nicole, Wachholz Kirsten, Eibl Hansjoerg, Hiddemann Wolfgang, Issels Rolf D
Department of Medicine III, University Hospital Grosshadern, Ludwig-Maximilians-University, Munich, Germany.
J Control Release. 2008 Jan 22;125(2):112-20. doi: 10.1016/j.jconrel.2007.10.009. Epub 2007 Oct 22.
Lysolipid-based thermosensitive liposomes have been successfully introduced as efficient drug delivery system with fast drug release upon heat treatment. Hexadecylphosphocholine (HePC) is structurally related to 1-palmitoyl-2-lyso-sn-glycero-3-phosphocholine (P-lyso-PC) but chemically and metabolically more stable, thereby offering strong antineoplastic, antiprotozoal and antifungal activity. We investigated the properties of HePC in low temperature sensitive (LTSL) and high temperature sensitive liposomes (HTSL) based on 1,2-dipalmitoyl-sn-glycero-3-phosphoglyceroglycerol (DPPGOG). Therefore liposomes composed of DPPC/DSPC=8:2 (molar ratio), DPPC/DSPC/DPPGOG=5:2:3, HePC/DPPC/DSPC/DPPGOG=1:4:2:3, HePC/DSPC/DPPGOG=1:6:3 and P-lyso-PC/DPPC/DSPC/DPPGOG=1:4:2:3 were prepared by the lipid film hydration and extrusion method and compared with regard to stability at 37 degrees C and the release kinetics under heating conditions (38 degrees C-45 degrees C) in the presence of fetal calf serum. Each formulation was characterized for size distribution, zeta-potential, encapsulation efficiency and phase transition temperature (T(m)). The influence of heat on the cytotoxic activity of HePC in TSL was investigated using the WST-1 assay in BFS-1 fibrosarcoma and C6 glioma cells for the low (HePC-LTSL, T(m)=42.9 degrees C) and the high (HePC-HTSL, T(m)=48.5 degrees C) temperature sensitive formulations and compared to micellar HePC or plain TSL. The cellular HePC uptake after 15 min incubation at 37 degrees C or 42 degrees C was determined by liquid chromatography tandem-mass spectrometry (LC-MS/MS). As expected, HePC increases the release rate of TSL similar to lysolipid. The cytotoxicity of HePC in TSL was heat inducible and stronger than the one induced by micellar HePC which did not respond to heat. A possible explanation for this raise in cytotoxicity of HePC in TSL is the increased cellular transfer of HePC under heating conditions demonstrated by LC-MS/MS.
基于溶血脂质的热敏脂质体已成功引入，作为一种高效的药物递送系统，在热处理时能快速释放药物。十六烷基磷酸胆碱（HePC）在结构上与1-棕榈酰-2-溶血-sn-甘油-3-磷酸胆碱（P-溶血-PC）相关，但在化学和代谢方面更稳定，因此具有强大的抗肿瘤、抗原生动物和抗真菌活性。我们研究了基于1,2-二棕榈酰-sn-甘油-3-磷酸甘油甘油（DPPGOG）的低温敏感（LTSL）和高温敏感脂质体（HTSL）中HePC的性质。因此，通过脂质膜水化和挤压法制备了由DPPC/DSPC = 8:2（摩尔比）、DPPC/DSPC/DPPGOG = 5:2:3、HePC/DPPC/DSPC/DPPGOG = 1:4:2:3、HePC/DSPC/DPPGOG = 1:6:3和P-溶血-PC/DPPC/DSPC/DPPGOG = 1:4:2:3组成的脂质体，并在胎牛血清存在的情况下，比较了它们在37℃下的稳定性以及加热条件（38℃-45℃）下的释放动力学。对每种制剂的粒径分布、zeta电位、包封率和相变温度（T(m)）进行了表征。使用WST-1测定法，在BFS-1纤维肉瘤和C6胶质瘤细胞中，研究了热对TSL中HePC细胞毒性活性的影响，针对低温（HePC-LTSL，T(m)=42.9℃）和高温（HePC-HTSL，T(m)=48.5℃）敏感制剂，并与胶束状HePC或普通TSL进行比较。通过液相色谱串联质谱（LC-MS/MS）测定在37℃或
