面两亲性分子改造的铂类脂质体的分泌性磷脂酶A<sub>2</sub>响应性和体外抗肿瘤活性

刘燕娇; 文成; 李丹; 高佩; 朱国东

doi:10.11665/j.issn.1000-5048.20220407

面两亲性分子改造的铂类脂质体的分泌性磷脂酶A₂响应性和体外抗肿瘤活性

五邑大学生物科技与大健康学院，江门 529030

基金项目: 国家自然科学基金资助项目（No.8167131086）

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出版历程
- 收稿日期: 2022-02-24
- 修回日期: 2022-06-01
- 刊出日期: 2022-08-24

Secretory phospholipase A₂ responsiveness and in vitro anti-tumor activity of oxaliplatin-loaded liposomes modified with facial amphiphiles

School of Biotechnology and Health Sciences, Wuyi University, Jiangmen 529030, China

Funds: This study was supported by the National Natural Science Foundation of China (No.8167131086)

摘要

摘要: 铂类脂质体在肿瘤部位的定点药物释放是其发挥疗效的关键，本研究以二棕榈酰磷脂酰胆碱和二硬脂酰磷脂酰乙醇胺-聚乙二醇2k两种磷脂为基础，构建带有面两亲性分子石胆酸（lithocholic acid， LCA）或3-酮石胆酸（3-keto lithocholic acid， kLCA）的新型脂质体（LCA-Lip，kLCA-Lip），测试其对肿瘤分泌性磷脂酶A₂ （secretory phospholipase A_2，sPLA₂）的响应性。采用薄膜水化-挤出法制备脂质体，对所制备脂质体进行理化性质表征，通过酶刺激脂质体释放荧光素CF实验研究其酶响应性，并通过体外毒性实验研究其抑制肿瘤细胞增殖的活性。结果表明：所制备的面两亲分子改造的奥沙利铂脂质体平均粒径约为100 nm，且分散均匀（PDI < 0.11），相比于不含面两亲性分子的脂质体（C-Lip），包封率和载药量没有显著性差异；与不加胎牛血清（FBS）的孵育条件相比，加入10%，50%的FBS没有显著性地增加奥沙利铂从脂质体的泄漏率。体外荧光素CF释放特性实验结果表明，相比C-Lip脂质体，LCA-Lip和kLCA-Lip脂质体对Colo205结肠癌细胞分泌的sPLA₂响应度更高，在酶的作用下LCA-Lip和kLCA-Lip在24 h释放约70%的荧光素CF，而C-Lip释放率仅约20%；体外抗肿瘤活性结果表明，奥沙利铂LCA-Lip和奥沙利铂kLCA-Lip对高表达sPLA₂的Colo205细胞增殖的抑制效果明显高于奥沙利铂C-Lip。本研究表明，LCA或者3-kLCA面两亲性分子可提高脂质体对肿瘤细胞分泌的sPLA₂的响应性，为未来开发可应用于临床，具有定点释药功能的铂类脂质体提供了新的思路。
- 分泌性磷脂酶A₂ /
- 触发释放 /
- 石胆酸 /
- 3-酮石胆酸 /
- 酶响应性脂质体 /
- 抗肿瘤活性 /
- 面两亲性分子
Abstract: Modulating drug release from liposomes at tumor sites are important for eliciting therapeutic effects of platinum drugs considering their low permeability through liposomal membranes, here a novel secretory phospholipase A₂ (sPLA₂) responsive-liposome system was constructed for oxaliplatin (L-OHP).Lipid ingredients dipalmitoyl phosphatidylcholine and distearoyl phosphoethanolamine-PEG2k, together with facial amphiphiles (FAs) including lithocholic acid (LCA) or 3-keto lithocholic acid (kLCA) were used to prepare sPLA₂responsive-liposome (LCA-Lip or kLCA-Lip) by thin-film hydration method.The physicochemical properties, sPLA₂-responsive drug release and anti-tumor activity were evaluated in vitro.The results indicated L-OHP loaded liposomes modified with FAs had similar particle sizes of approximately 100 nm and narrow size distributions (PDI < 0.11).Compared with non-FAs-containing liposomes (C-Lip), LCA-Lip or kLCA-Lip has a comparable entrapment efficiency and loading efficiency.LCA-Lip or kLCA-Lip didn't show significant higher drug leakage at the presence of 10% or 50% fetal bovine serum (FBS) in media than that in media without FBS.Treated with secretory phospholipase A₂ from Colo205 cells culture conditioned medium (CCM sPLA₂) for 24 h, FAs modified liposomes released about 70% of carboxyfluorescein (CF), while C-Lip only released 20% of CF.Compared to L-OHP loaded C-Lip, L-OHP-loaded FAs-included formulations had much greater anti-proliferative activity against sPLA₂-secreting Colo205 cells.In summary, our results shows that LCA or kLCA promotes responsiveness of liposomes to tumor-related sPLA₂ and points to a new way to develop platium drugs-loaded liposomal delivery systems with better release mechanisms.
- secretory phospholipase A₂ /
- triggered-release /
- lithocholic acid /
- 3-keto lithocholic acid /
- enzyme-responsive liposome /
- anti-tumor activity /
- facial amphiphiles

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面两亲性分子改造的铂类脂质体的分泌性磷脂酶A2响应性和体外抗肿瘤活性

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Secretory phospholipase A2 responsiveness and in vitro anti-tumor activity of oxaliplatin-loaded liposomes modified with facial amphiphiles

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面两亲性分子改造的铂类脂质体的分泌性磷脂酶A₂响应性和体外抗肿瘤活性

Secretory phospholipase A₂ responsiveness and in vitro anti-tumor activity of oxaliplatin-loaded liposomes modified with facial amphiphiles