榄香烯增强卡巴他赛抗胶质瘤作用考察及其双靶向阳离子脂质体的制备优化与体外药效学评价  

Elemene Enhances Anti-glioma Effect of Cabazitaxel and Its Dual-targeted Cationic Liposome Preparation Optimization and in Vitro Pharmacodynamic Evaluation

作　　者：尹丽燕 王蓉蓉[2] 李杰[3] 谢恬 YIN Liyan;WANG Rongrong;LI Jie;XIE Tian(School of Traditional Chinese Medicine(TCM),Guangdong Pharmaceutical University,Guangzhou 511400,China;School of Pharmacy,Hangzhou Normal University,Hangzhou 311100,China;Chengdu University of TCM,Chengdu 611130,China)

机构地区：[1]广东药科大学中药学院,广州511400 [2]杭州师范大学药学院,杭州311100 [3]成都中医药大学,成都611130

出　　处：《中国实验方剂学杂志》2024年第12期173-184,共12页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81730108);四川省科技计划项目(24NSFSC2472);浙江省科技计划基金项目(2021C03087)。

摘　　要：目的:探究榄香烯(ELE)增强卡巴他赛(CTX)抗胶质瘤疗效,制备一种共载ELE/CTX的双靶向阳离子脂质体(LIP)用于胶质瘤的治疗研究,并达到增加药效、减少不良反应的作用,经体外药效学试验揭露其优势及作用机制。方法:采用高速剪切法联合探头超声法制备脂质体ELE/CTX@LIP,并采用纳米粒径电位仪对其粒径及电位进行表征,采用高效液相色谱法(HPLC)测定ELE/CTX包封率及载药量;采用细胞增殖与活性检测法(CCK-8)检测ELE/CTX体外细胞增殖抑制活性;通过JMP Pro 16软件,以包封率为指标优化脂质体(ELE/CTX@LIP)工艺参数;通过体外细胞增殖抑制活性和体外细胞摄取,筛选最佳阳离子材料种类、含量与配比,在此基础上制备双靶向阳离子脂质体并表征其形态、粒径稳定性等,验证其对RG2胶质瘤细胞的细胞周期和细胞凋亡的影响。结果:ELE/CTX抗胶质瘤活性结果显示ELE/CTX分别对C6、RG2细胞具有更强的细胞增殖抑制活性;体外细胞增殖抑制活性和体外细胞摄取结果显示阳离子材料用量为总含量的0.10%;T7、精氨酸-甘氨酸-天冬氨酸三肽序列(cRGD)与磷脂最佳配比为1∶1∶50。T7/cRGD-ELE/CTX@CLIP[1,2-二油酰氧基-3-二甲基氨基丙烷(DlinMC3-DMA)]、T7/cRGD-ELE/CTX@CLIP[二肉豆蔻酰甘油-聚乙二醇2000(DMG-PEG2000)]呈现多层次球状纳米结构,粒径分别为146.0 nm和111.3 nm,血清稳定性良好;体外细胞增殖抑制结果显示,与单靶向脂质体或双靶向非阳离子脂质体比较,T7/cRGD-ELE/CTX@CLIP在体外对胶质瘤细胞具有更高的细胞增殖抑制活性;T7/cRGD-ELE/CTX@CLIP影响胶质瘤细胞凋亡及周期,结果显示,ELE/CTX联用能通过脂质体载体能更加有效的激活细胞凋亡通道抑制胶质瘤细胞增殖,且使用T7/cRGD短肽及阳离子修饰后,增强了细胞凋亡诱导能力,ELE/CTX能有效将胶质瘤细胞周期阻滞于G2/M期,且经T7/cRGD靶向修饰后,效果增强。结论:ELE可以增强CTX抗胶�Objective::To investigate whether elemene(ELE)enhances the anti-glioma efficacy of cabazitaxel(CTX),and prepare a double-targeted cationic liposome(LIP)co-loaded with ELE/CTX for the treatment of glioma,and to achieve the effect of increasing the efficacy and reducing the adverse reactions.Pharmacodynamic tests in vitro were performed to explore the advantages and mechanism of its preparation.M ethod::ELE/CTX@LIP was prepared by high speed shear combined with probe ultrasound,the particle size and potential were characterized by nano-particle size potentiometer,and high performance liquid chromatography(HPLC)was used to determine the encapsulation efficiency and drug loading capacity of CTX/ELE.The cytotoxicity of ELE/CTX in vitro was detected by cell proliferation and activity assay(CCK8).JMP Pro 16 software was used to optimize the process parameters of ELE/CTX@LIP based on encapsulation efficiency.The optimal cationic material type,content and ratio were screened by in vitro cytotoxicity and in vitro cell uptake,on this basis,the dual-targeted cationic liposome T7/arginine glycine aspartate tripeptide sequence(T7/cRGD)-ELE/CTX@CLIP was prepared,the stability of morphology and particle size were characterized,and the effect of T7/cRGD-ELE/CTX@CLIP on the apoptosis inducing ability and cell cycle regulation ability of glioma cells was analyzed by cell cycle and apoptosis.Result::ELE/CTX showed stronger anti-glioma activity on C6 and RG2 cells.The results of in vitro cytotoxicity and in vitro cell uptake showed that the amount of cationic material was 0.10%of the total content.The optimum ratio of T7,cRGD and phospholipids was 1∶1∶50.T7/cRGD-ELE/CTX@CLIP[1,2-dilinoleyloxy-3-dimethylaminopropane(Dlin-MC3-DMA)]and T7/cRGD-ELE/CTX@CLIP[1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol 2000(DMG-PEG2000)]showed multi-level spherical nanostructures with particle sizes of 146.0,111.3 nm,respectively,and were stable in serum.In vitro cytotoxicity results showed that T7/cRGD-ELE/CTX@CLIP had higher cytotoxicity

关 键 词：双靶向 阳离子脂质体 脑胶质瘤 榄香烯 卡巴他赛 增效 工艺优化 

分 类 号：R22[医药卫生—中医基础理论] R94[医药卫生—中医学] R28R979.1