目的 制備單甲氧基聚乙二醇5000-聚己內(nèi)酯10000（mPEG5000-PCL10000）修飾的多西他賽（DTX）脂質(zhì)體（DTXPLip），并初步評價(jià)其體外抗腫瘤活性。方法 采用薄膜分散-水化法制備DTX-Plip。以粒徑、包封率（EE）、載藥量（LC）為評價(jià)指標(biāo)，通過單因素及正交試驗(yàn)優(yōu)選DTX-PLip的最佳處方工藝；研究DTX-PLip的透射電鏡微觀形態(tài)、粒徑、ζ電位、EE及LC；考察其在4℃放置21 d內(nèi)的穩(wěn)定性；采用透析法對DTX-PLip的體外釋放特性進(jìn)行研究；采用MTT法評估DTXPLip對小鼠乳腺癌4T1細(xì)胞的增殖抑制效應(yīng)；通過體外細(xì)胞攝取實(shí)驗(yàn)，結(jié)合熒光顯微鏡觀察與流式細(xì)胞術(shù)定量分析4T1細(xì)胞對DTX-PLip的攝取效率。結(jié)果 DTX-PLip最佳處方為mPEG5000-PCL10000用量150 mg，DTX用量8 mg，藥脂比為1∶ 20，膽脂比為1∶ 5。透射電子顯微鏡圖片顯示DTX-PLip具封閉囊泡結(jié)構(gòu)，平均粒徑為（82.13±3.33） nm，ζ電位為（-15.70±3.86） mV； EE為（89.34±1.07）%，LC為（2.04±0.02）%；體外釋放結(jié)果表明，DTX-PLip 72 h體外累積釋放率為58%，具有一定的緩釋性，4℃放置21 d內(nèi)儲存穩(wěn)定；細(xì)胞毒實(shí)驗(yàn)結(jié)果表明DTX-PLip對4T1細(xì)胞的半數(shù)抑制濃度（IC50）值為（0.13±0.01） μg·mL^-1，均顯著高于DTX溶液組和DTX脂質(zhì)體（DTX-Lip）組（P＜0.001） ，具有良好的抗腫瘤活性；熒光顯微鏡觀察結(jié)果表明DTX-PLip在各個(gè)時(shí)間平均熒光強(qiáng)度均顯著高于DTX-Lip（P＜0.01），流式細(xì)胞術(shù)結(jié)果表明，與DTX-Lip相比，DTX-PLip細(xì)胞攝取量顯著增加（P＜0.001）。結(jié)論 DTX-PLip表現(xiàn)出明顯的緩釋作用，能增強(qiáng)DTX的抗腫瘤效果。

Objective To prepare monomethoxy polyethylene glycol 5000- Polycaprolactone 10000 (mPEG5000-PCL10000) modified docetaxel (DTX) liposomes (DTX-PLip), preliminary evaluate the antitumor activity in vitro. Methods DTX-PLip was prepared by thin film dispersion-hydration method. The optimal prescription process of DTX-PLip was optimized by one-way and orthogonal tests using particle size, encapsulation efficiency (EE), and drug loading capacity (LC) as evaluation indexes, the transmission electron microscopy microstructure, particle size, ζ potential, EE, LC, and stability of the optimized DTX-PLip were investigated. The in vitro release characteristics of DTX-PLip were studied using dialysis method. MTT method was used to evaluate the inhibitory effect of DTX-PLip on the proliferation of mouse breast cancer 4T1 cells. Quantitative analysis of the uptake efficiency of DTX PLip by 4T1 cells was conducted through in vitro cell uptake experiments, combined with fluorescence microscopy observation and flow cytometry. Results The optimal formulation of DTX-PLip was mPEG5000-PCL10000 at 150 mg, DTX at 8 mg, with a drug-to-lipid ratio of 1∶20 and a bile-to-lipid ratio of 1∶5. Transmission electron microscopy images showed that DTX-PLip had a closed vesicular structure, with an average particle size of (82.13 ±3.33) nm and a ζ potential of (-15.70 ±3.86) mV. EE was (89.34 ±1.07)%, and LCwas (2.04 ±0.02)%. In vitro release results indicated that the cumulative release rate of DTX-PLip was 58% after 72 h, demonstrating a certain sustained-release property. It remained stable during storage at 4 ℃ for 21 d. The cytotoxicity assay results showed that the IC₅₀ value of DTX-PLip against 4T1 cells was (0.13 ±0.01) μg·mL^-1, which was significantly higher than that of the DTX solution group and the DTX-Lip group (P <0.001), indicating good anti-tumor activity. Fluorescence microscopy observations revealed that the average fluorescence intensity of DTX-PLip was significantly higher than that of DTX-Lip at each time point (P <0.01). Flow cytometry results indicated that the cellular uptake of DTX-PLip was significantly increased compared with DTX-Lip (P <0.001). Conclusion DTX-PLip demonstrated a significant sustained-release effect and could enhance the anti-tumor efficacy of DTX.

R979.1

河南省科技攻關(guān)項(xiàng)目（242102310563）；河南省科技攻關(guān)項(xiàng)目（ 232102310337）；新鄉(xiāng)醫(yī)學(xué)院三全學(xué)院生物與醫(yī)藥省級重點(diǎn)學(xué)科項(xiàng)目資助（ ZDXKXM019）；新鄉(xiāng)醫(yī)學(xué)院三全學(xué)院優(yōu)秀青年教師培養(yǎng)計(jì)劃（ SQ2023YQJH06）