目的 制備西瑞香素納米混懸劑，并考察其對多種腫瘤細胞增殖的抑制作用。方法 以粒徑、Zeta電位、多分散性指數(shù)（PDI）為指標(biāo)，考察穩(wěn)定劑、藥載比、超聲溫度、均質(zhì)溫度、均質(zhì)次數(shù)對西瑞香素納米混懸劑反溶劑沉淀法制備的影響，優(yōu)化最佳處方和工藝條件。觀察西瑞香素納米混懸劑的透射電鏡表征，并進行X射線衍射、差示掃描量熱分析，考察其在血漿、PBS中的穩(wěn)定性以及載藥量、體外釋放情況，采用MTT法比較其對BT474、SKBR-3、A549、HeLa、HepG2細胞的體外細胞毒性。結(jié)果 最佳處方和工藝條件：TPGS為穩(wěn)定劑，藥載比1∶1，共同溶解于DMSO中，在超聲（25℃，250 W）條件下，緩慢滴注于去離子水中，透析除去有機溶劑，高壓均質(zhì)（25℃，150 MPa）20次，即得西瑞香素納米混懸劑，其平均粒徑為（163.1±5.4）nm，Zeta電位為（-11.4±0.7）mV，PDI為0.15±0.04。西瑞香素納米混懸劑近乎為球形，大小較均勻；在血漿中穩(wěn)定存在，不存在溶血現(xiàn)象，滿足靜脈注射需求，平均載藥量為（39.16±1.09）%。西瑞香素納米混懸劑對5種受試細胞的生長抑制作用顯著提高，并呈現(xiàn)劑量相關(guān)性，尤其是對SKBR-3、A549、HeLa、HepG2細胞，IC₅₀在2.1～3.4 μg/mL。結(jié)論 西瑞香素納米混懸劑具有小粒徑、高載藥量、顯著腫瘤細胞毒性等優(yōu)點，在抗腫瘤研究方面具有良好的應(yīng)用前景。

objective To prepare Daphnoretin Nanosuspensions (Dan-NSps) and study its anti-tumor activities in vitro. Methods Dan-NSps were prepared by anti-solvent precipitation method with particle size, Zeta potential, and PDI as indexes, and stabilizer, drug loading ratio, ultrasonic temperature, homogeneous temperature, and homogeneous times were studied to optimize the optimum prescriptions and technological conditions. TEM of Dan-NSps was observed, and X-ray diffraction and differential scanning calorimetry were carried out. Stabilities in plasma and PBS were studied, and the drug loadings and releases in vitro were also compared. MTT assay was used to evaluate the in vitro cytotoxicities against BT474, SKBR-3, A549, HeLa, and HepG2 cells. Results The best preparation method was as following:TPGS was as stabilizer with drug loading ratio 1:1, and was dissolved in DMSO together. Under the condition of ultrasonic (25 W, 250 W), it was slowly dripped into deionized water, and the organic solvent was removed by dialysis. The high pressure homogenization (25℃, 150 MPa) was 20 times. The average particle size of Dan-NSps was (163.1 ±5.4) nm, the Zeta potential was (−11.4 ±0.7) mV, and PDI was 0.15 ±0.04. Naonosuspensions were nearly spherical, and more uniform in size. Dan-NSps remained stable in plasma with no hemolysis, meeting the demand of intravenous administration. The drug loading content was determined to be as high as (39.16 ±1.09)%. MTT assay showed that Dan-NSps had higher cytotoxicities against 5 kinds of cell lines with a dose dependent relationship, especially for SKBR-3, A549, HeLa, and HepG2 cells, which IC₅₀ were at 2.1 to 3.4 g/mL. Conclusion Dan-NSps had advantages of small particle size, high drug load, and significant tumor cell toxicity, which has a good application prospect in the field of anti-tumor research.

國家自然科學(xué)基金-廣東聯(lián)合基金資助項目（U1401223）