RPV修饰的紫杉醇与五味子乙素脂质体处方工艺优化及体外抗肿瘤活性初步评价

张璐 何思雨 刘昕泽 孔亮 李学涛

中图分类号 R943;R285 文献标志码 A 文章编号 1001-0408（2021）10-1173-08

DOI 10.6039/j.issn.1001-0408.2021.10.04

摘 要 目的：制备细胞穿膜肽RPV修饰的紫杉醇与五味子乙素脂质体，并初步评价其体外抗肿瘤活性。方法：采用薄膜分散法制备RPV修饰的紫杉醇与五味子乙素脂质体。以胆固醇加入量、紫杉醇加入量和探头超声时间间隔为考察因素，紫杉醇和五味子乙素两药的平均包封率為考察指标，通过Box-Benhken设计-响应面实验优化RPV修饰的紫杉醇与五味子乙素脂质体的处方工艺，并对最优处方工艺所制脂质体进行表征。采用磺酰罗丹明B染色法考察RPV修饰的空白脂质体、紫杉醇与五味子乙素脂质体、RPV修饰的紫杉醇与五味子乙素脂质体对人卵巢癌细胞SK-OV-3的体外毒性作用，并采用细胞划痕实验和Transwell小室侵袭实验分别考察上述3种脂质体对SK-OV-3细胞迁移、侵袭能力的影响。结果：最优处方工艺为磷脂44 mg、胆固醇8 mg、紫杉醇0.64 mg、五味子乙素1.5 mg，探头超声时间间隔5 s，总处方量为5 mL。根据最优处方工艺制得的脂质体为类球形，粒径为（126.49±1.19）nm，Zeta电位为（-4.83±0.61） mV，脂质体中两药的平均包封率为（93.88±1.67）%。与RPV修饰的空白脂质体比较，经紫杉醇与五味子乙素脂质体、RPV修饰的紫杉醇与五味子乙素脂质体处理后，SK-OV-3细胞的存活率、迁移抑制率和侵袭率均显著降低（P<0.05），且RPV修饰的紫杉醇与五味子乙素脂质体的作用优于紫杉醇与五味子乙素脂质体（P<0.05）。结论：成功制备了RPV修饰的紫杉醇与五味子乙素脂质体，且其具有一定的体外抗肿瘤活性。

关键词 紫杉醇;五味子乙素;细胞穿膜肽;脂质体;Box-Benhken设计-效应面实验;处方工艺优化;抗肿瘤活性;人卵巢癌细胞SK-OV-3

Prescription Technology Optimization of RPV Modified Paclitaxel and Schisandrin B Liposomes and Preliminary Evaluation of Antitumor Activity in vitro

ZHANG Lu，HE Siyu，LIU Xinze，KONG Liang，LI Xuetao（School of Pharmacy， Liaoning University of TCM， Liaoning Dalian 116600， China）

ABSTRACT   OBJECTIVE： To prepare paclitaxel and schisandrin B liposomes modified by cell penetrating peptide RPV， and to preliminarily evaluate its anti-tumor activity in vitro. METHODS： RPV modified paclitaxel and schisandrin B liposomes were prepared by film dispersion method. Box-Benhken design-response surface methodology was used to optimize the prescription technology of RPV modified paclitaxel and schisandrin B liposomes using the amount of cholesterol and paclitaxel， the time interval of ultrasound probe as factors， average entrapment efficiency of paclitaxel and schisandrin B was used as the index. The liposomes prepared by the optimal technology were characterized. Sulfonylrhodamine B staining method was used to investigate in vitro toxicity of RPV modified blank liposomes， paclitaxel and schisandrin B liposomes， RPV modified paclitaxel and schisandrin B liposomes to human ovarian cancer cell SK-OV-3. The effects of 3 kinds of liposomes on the migration and invasion ability of SK-OV-3 cells were investigated by cell scratch test and Transwell chamber invasion test. RESULTS： The optimal prescription technology was phospholipid 44 mg， cholesterol 8 mg， paclitaxel 0.64 mg， schisandrin B 1.5 mg， ultrasonic probe time interval 5 s， prescription dosage 5 mL. According to the optimal prescription technology， the liposomes were spherical in shape， and the particle size was （126.49±1.19） nm， Zeta-potential was （-4.83±0.61） mV， average entrapment efficiency of liposomes was （93.88±1.67）%. Compared with RPV modified blank liposomes， after treated with paclitaxel and schisandrin B liposomes and RPV modified paclitaxel and schisandrin B liposomes， the survival rate， migration inhibition rate and invasion rate of SK-OV-3 cells were significantly decreased （P<0.05）. The effects of RPV modified paclitaxel and schisandrin B liposomes was better than those of paclitaxel and schisandrin B liposomes （P<0.05）. CONCLUSIONS： RPV modified paclitaxel and schisandra B liposome are successfully prepared， and they have certain antitumor activity in vitro.

KEYWORDS   Paclitaxel; Schisandrin B; Cell penetrating peptide; Liposome; Box-Benhken design-response surface methodology; Prescription technology optimization; Anti- tumor activity; SK-OV-3 cells

卵巢癌作为妇科生殖系统的三大恶性肿瘤之一，致死率较高，目前临床上常采用紫杉醇相关制剂进行治疗[1]。……