温度敏感性脂质体制剂是改善药物治疗指数且毒性最小的有前途的工具。这项研究的目的是研究伴随递送含有热敏脂质体(TSLs)的顺铂(Cis)和多柔比星(Dox)的潜力,并在体外和体内进行轻度高温抗癌。聚乙二醇涂层DPPC/DSPC,热敏和DSPC,制备并表征掺入Cis和Dox的非热敏脂质体。采用常规差示扫描量热法(DSC)和傅里叶变换红外光谱(FT-IR)技术研究了药物-磷脂的相互作用和相容性。在高温条件下,在苯并[a]芘(BaP)诱导的纤维肉瘤中评估了这些制剂的化疗功效。测得所制备的热敏脂质体的尺寸直径为120±10nm。DSC数据显示DSPC+Dox和DSPC+Cis曲线的变化,同时比较纯DSPC和药物。然而,FITR分别和混合物中的磷脂和药物谱相同。数据显示Cis-Dox-TSL的更高效力,因为在高热条件下的该组动物中记录到84%的肿瘤生长抑制。卡普兰-梅尔曲线显示,高温下Cis-Dox-TSL和无高温下Cis-Dox-NTSL处理组的动物存活率分别为100%和80%,分别。然而,Cis-TSL和Dox-TSL表现出50%的存活率,而用Dox-NTSL和Cis-NTSL治疗的动物组仅记录了20%的存活率。流式细胞术分析显示Cis-Dox-NTSL增强了肿瘤细胞凋亡的诱导,记录为18%。不出所料,Cis-Dox-TSL显示出巨大的潜力,因为39%的细胞被测量为凋亡细胞,与Cis-Dox-NTSL相比,Dox-TSL和Cis-TSL。通过流式细胞术进行的细胞凋亡分析清楚地表明在施用Cis-Dox-TSL制剂时治疗期间热疗的影响。最后,通过共聚焦显微镜对肿瘤组织进行的免疫组织化学分析显示,在Sham-NTSL和Sham-TSL的载体治疗的动物中,pAkt的表达增加了几倍。然而,Cis-Dox-TSL在Akt的表达中表现出极大的降低,下降了11倍。本研究的结果指导了在高温条件下伴随递送阿霉素和顺铂的热敏脂质体在开发治疗癌症的新型治疗策略中的作用。
The temperature sensitive liposomal formulations are a promising tool to improve the therapeutic index of the drugs with minimal toxicity. The aim of this study was to investigate the potential of concomitant delivery of cisplatin (Cis) and doxorubicin (Dox) containing thermosensitive liposomes (TSLs) with mild hyperthermia against cancer in vitro and in vivo. The polyethylene glycol coated DPPC/DSPC, thermosensitive and DSPC, non-thermosensitive liposomes incorporating Cis and Dox were prepared and characterized. A conventional Differential Scanning Calorimetry (DSC) technique and Fourier Transform Infrared Spectroscopy (FT-IR) were applied to study drug-phospholipid interaction and compatibility. The chemotherapeutic efficacy of these formulations was evaluated in benzo[a]pyrene (BaP) induced fibrosarcoma under hyperthermic condition. The size diameter of prepared thermosensitive liposomes was measured to be 120 ± 10 nm. The DSC data exhibited the changes in the curves of DSPC + Dox and DSPC + Cis while comparing the pure DSPC and drugs. However, the FITR showed same spectrum of phospholipids and drugs individually and in the mixture as well. The data showed higher efficacy of Cis-Dox-TSL as 84% inhibition in tumor growth was recorded in this group of animals in hyperthermic condition. The Kaplan-Meir curve revealed, 100% and 80% survival of the animals in the groups treated with Cis-Dox-TSL under hyperthermia and Cis-Dox-NTSL without hyperthermia, respectively. However, Cis-TSL as well as Dox-TSL exhibited 50% survival, while only 20% survival was recorded in the groups of animals treated with Dox-NTSL and Cis-NTSL. The flow cytometry analysis revealed that Cis-Dox-NTSL augments the induction of apoptosis in the tumor cells which was recorded as 18%. As expected, Cis-Dox-TSL showed great potential as 39% of cells were measured as apoptotic cells, significantly very high in comparison to Cis-Dox-NTSL, Dox-TSL and Cis-TSL as well. The apoptotic analysis of the cells by flow cytometry clearly indicated the effect of hyperthermia during the treatment while Cis-Dox-TSL formulation was administered. Finally, the immunohistochemical analysis of the tumor tissues by confocal microscopy exhibited several fold increases in the expression of pAkt in the animals treated with vehicles in Sham-NTSL as well as Sham-TSL. However, Cis-Dox-TSL showed great reduction in the expression of Akt, as it declined by 11-fold. The results of the present study directed the role of concomitant delivery doxorubicin and cisplatin containing thermosensitive liposomes under hyperthermic conditions for the development of a novel therapeutic strategy for the treatment of cancer.