细胞穿膜肽PFV修饰紫杉醇/青蒿琥酯共载靶向胶束的制备及体外抗肿瘤作用研究  

Study on Preparation and in vitro Anticancer Activity of Cell Penetrating Peptide PFV-modified Paclitaxel/artesunate Co-loaded Targeting Micelles

作　　者：王为 李学涛[1] 孔亮[1] 姜爽 罗一夫 王晓波[1,2] WANG Wei;LI Xuetao;KONG Liang;JIANG Shuang;LUO Yifu;WANG Xiaobo(School of Pharmacy,Liaoning University of TCM,Liaoning Dalian 116600,China;No.967 Hospital of the Joint Logistic Support Force of PLA,Liaoning Dalian 116011,China)

机构地区：[1]辽宁中医药大学药学院,辽宁大连116600 [2]解放军联勤保障部队第九六七医院药物研究所,辽宁大连116011

出　　处：《中国药房》2020年第21期2592-2597,共6页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81973462,No.81874347)。

摘　　要：目的:制备细胞穿膜肽PFV修饰紫杉醇(PTX)/青蒿琥酯(ART)共载靶向胶束,并考察其体外抗肿瘤活性。方法:按前期优化的工艺,采用薄膜水化法制备PFV修饰PTX/ART共载靶向胶束,并对其进行表征。以空白胶束作为空白对照,采用磺基罗丹明B法评价PTX胶束、ART胶束、PTX/ART胶束以及PFV修饰PTX/ART共载靶向胶束对人胃癌BGC-823细胞的毒性;以香豆素作为荧光探针取代PTX,制成相应的不同胶束,采用流式细胞仪和荧光显微镜测定和观察BGC-823细胞对各胶束的摄取情况和靶向性;并通过Transwell小室法考察PTX胶束、ART胶束、PTX/ART胶束以及PFV修饰PTX/ART共载靶向胶束对BGC-823细胞侵袭的影响。结果:PFV修饰PTX/ART共载靶向胶束的平均粒径为(51.30±3.95)nm,分散系数为0.19±0.01,Zeta电位为(0.21±0.02)mV,PTX、ART的包封率均高于90%,形态呈圆球形。空白胶束对BGC-823细胞无明显毒性,PTX胶束、PTX/ART胶束以及PFV修饰PTX/ART共载靶向胶束对BGC-823细胞的半数抑制浓度分别为(3.09±0.22)、(1.93±0.24)、(1.11±0.15)μmol/L;不同胶束在BGC-823细胞核中的分布数量排序依次为PFV修饰香豆素/ART胶束>香豆素/ART胶束>香豆素胶束>空白对照,抑制细胞侵袭作用的大小依次为PFV修饰PTX/ART共载靶向胶束>PTX/ART胶束>ART胶束>PTX胶束>空白对照。结论:制备的PFV修饰PTX/ART共载靶向胶束符合《中国药典》要求;其对BGC-823细胞具有较强的细胞毒性,可提高药物的靶向性和细胞对药物的摄取能力,并能抑制肿瘤细胞的侵袭和转移。OBJECTIVE:To prepare cell penetrating peptide PFV-modified paclitaxel(PTX)/artesunate(ART)co-loaded targeting micelles,and to investigate in vitro anti-tumor activity.METHODS:According to optimal technology,PFV-modified PTX/ART co-loaded targeting micelles were prepared by membrane hydration method,and were characterized.Using blank micelle as blank control,sulforhodamine B(SRB)method was used to evaluate the toxicity of PTX micelles,ART micelles,PTX/ART micelles and PFV-modified PTX/ART co-loaded targeting micelles to human gastric cancer BGC-823 cells.The coumarin was used as fluorescent probe replacing PTX to prepare corresponding micelles.Then,the uptake of BGC-823 cells to corresponding micelles and targeting effect were observed and determined by flow cytometry and fluorescence microscope.The effects of PTX micelles,ART micelles,PTX/ART micelles and PFV-modified PTX/ART co-loaded targeting micelles on the invasion of BGC-823 cells were investigated by Transwell chamber method.RESULTS:Average particle size of PFV-modified PTX/ART co-loaded targeting micelles was(51.30±3.95)nm;PDI was 0.19±0.01,and Zeta potential was(0.21±0.02)mV.The encapsulation efficiency of PTX and ART were higher than 90%.The shape of micelles were spherical.The blank micelles had no obvious toxicity to BGC-823 cells.The IC50 value of PTX micelles,PTX/ART micelles and PFV-modified PTX/ART co-loaded targeting micelles to BGC-823 cells were(3.09±0.22),(1.93±0.24),(1.11±0.15)μmol/L,respectively.The distribution amount of different micelles in BGC-823 cell nucleus in the descending order were PFV-modified coumarin/ART micelles>coumarin/ART micelles>coumarin micelles>blank control.The order of inhibitory effect was PFV-modified PTX/ART co-loaded targeting micelles>PTX/ART micelles>ART micelles>PTX micelles>blank control.CONCLUSIONS:Prepared PFV-modified PTX/ART co-loaded targeting micelles are in line with the quality of Chinese Pharmacopoeia.It shows strong cytotoxicity to BGC-823 cells,can improve the drug targeting and the cell upta

关 键 词：细胞穿膜肽 紫杉醇 青蒿琥酯 胶束 人胃癌BGC-823细胞 靶向作用 抗肿瘤作用 

分 类 号：R94[医药卫生—药剂学]