多西他賽PELGE納米粒的處方優(yōu)化、表征及其體外釋放和抗腫瘤活性的初步評價(jià)

廖龍飛 楊青青 漆婷婷 邱悅 肖洪濤

中圖分類號 R943;R944.9 文獻(xiàn)標(biāo)志碼 A 文章編號 1001-0408（2021）20-2492-07

DOI 10.6039/j.issn.1001-0408.2021.20.10

摘 要 目的：優(yōu)化多西他賽（DTX）-聚乙二醇-（聚乳酸-羥基乙酸）-聚乙二醇三嵌段共聚物（PELGE）-納米粒（NPs）的處方，對其進(jìn)行表征，并評價(jià)其體外釋放特性及抗腫瘤活性。方法：采用開環(huán)共聚法合成PELGE，采用乳化溶劑揮發(fā)法制備DTX-PELGE- NPs;采用高效液相色譜法測定DTX-PELGE-NPs中DTX的含量;以DTX用量、PELGE用量、泊洛沙姆188濃度為自變量，包封率為因變量，采用Box-Behnken設(shè)計(jì)-響應(yīng)面法優(yōu)化處方;采用激光粒度儀和透射電鏡測定DTX-PELGE-NPs的粒度和Zeta電位;以DTX注射液為參照，采用離心超濾法測定體外釋放率;以DTX注射液不含DTX的PELGE-NPs為參照，采用噻唑藍(lán)法考察體外細(xì)胞毒性。結(jié)果：最優(yōu)處方為DTX用量2.80 mg，PELGE用量20.60 mg，泊洛沙姆188濃度6%。優(yōu)化所得DTX-PELGE-NPs的包封率為（86.79±1.32）%，載藥量為（10.21±0.78）%，平均粒度為（78.4±2.9）nm，多分散性指數(shù)（PDI）為（0.187±0.018），Zeta電位為（-20.6±1.5）mV;電鏡下DTX-PELGE-NPs呈類球形，分布均勻。相較于DTX注射液（4 h時(shí)的累積釋放率約為92.3%），DTX-PELGE-NPs有明顯的緩釋效果（36 h時(shí)的累積釋放率約為78.6%）。0.1～50 μg/mL的PELGE-NPs對人乳腺癌細(xì)胞MCF-7無明顯細(xì)胞毒性（P>0.05），而0.5～10 μg/mL的 DTX-PELGE-NPs能夠顯著抑制人乳腺癌細(xì)胞MCF-7的生長，且作用（DTX-PELGE- NPs 10 μg/mL組除外）顯著強(qiáng)于同濃度DTX注射液（P<0.05）。結(jié)論：優(yōu)化所得處方工藝穩(wěn)定、可行;所得DTX-PELGE-NPs粒度均勻，包封率較高，緩釋效果明顯，體外抗腫瘤活性強(qiáng)于DTX注射液。

關(guān)鍵詞 多西他賽;聚乙二醇-（聚乳酸-羥基乙酸）-聚乙二醇三嵌段共聚物;Box-Behnken設(shè)計(jì)-響應(yīng)面法;高效液相色譜法;處方優(yōu)化

Formulation Optimization and Characterization of Docetaxel PELGE Nanoparticles and Preliminarily Evaluation of Its Drug Release and Antitumor Activity in vitro

LIAO Longfei1，YANG Qingqing2，QI Tingting1，QIU Yue1，XIAO Hongtao1（1. Dept. of Clinical Pharmacy， Sichuan Cancer Hospital/Sichuan Cancer Prevention and Control Center/the Affiliated Cancer Hospital of University of Electronic Science and Technology of China， Chengdu 610041， China; 2. Patent Examination Cooperation Sichuan Center， the Patent Office of CNIPA， Chengdu 610200， China）

ABSTRACT? ?OBJECTIVE： To optimize the formulation of docetaxel （DTX）-mPEG-PLGA-mPEG （PELGE）-nanoparticles （NPs）， and to characterize it and evaluate its in vitro drug release and antitumor activity. METHODS： PELGE were synthesized by ring-opening polymerization. DTX-PELGE-NPs were prepared by using emulsion solvent evaporation method. The content of DTX in DTX-PELGE-NPs was determined by HPLC. Box-Behnken design-response surface methodology was applied to optimize the formulation of the nanoparticles using the amount of DTX， PELGE and poloxamer 188 as independent variable， using entrapped efficiency as dependent variable. The particle size and Zeta-potential of DTX-PELGE-NPs were characterized by laser particle size analyzer and transmission electron microscope. The in vitro release of the DTX-PELGE-NPs was investigated by ultra-filtered centrifugation， using DTX injection as reference. In vitro cytotoxicity of the DTX-PELGE-NPs was investigated by MTT assay， using DTX and PELGE-NPs without DTX as reference. RESULTS： The optimal formulation included 2.80 mg DTX， 20.60 mg PELGE and 6% poloxamer 188. The entrapped efficiency of optimized DTX-PELGE-NPs was （86.79±1.32）%; drug-loading amount was （10.21±0.78）%， and average particle size was （78.4±2.9）nm; polydispersity coefficient was （0.187±0.018） and Zeta potential was （-20.6±1.5） mV. Furthermore， DTX- PELGE-NPs showed a regular spherical and uniform distribution under scanning electron microscopy. Compared with DTX injection （accumulative release rate of 92.3% at 4 h）， DTX- PELGE-NPs had a significant sustained-release effect （accumu- lative release rate of 78.6% at 36 h）. 0.1-50 μg/mL PELGE-NPs had no obvious cytotoxicity to human breast cancer cells MCF-7 （P>0.05）. 0.5-10 μg/mL DTX-PELGE-NPs could significantly inhibit the growth of human breast cancer cells MCF-7， and its inhibitory effect （except for DTX-PELGE-NPs 10 μg/mL group） was significantly stronger than that of DTX injection （P<0.05）. CONCLUSIONS： The optimized formulation is stable and feasible. The obtained DTX-PELGE-NPs not only have uniform particle size， high encapsulation rate obvious slow-release effect， but also have stronger anti-tumor effect in vitro than DTX injection.