目的 制備番荔素亞微乳，并進(jìn)行體內(nèi)外抗腫瘤作用研究，為番荔素的口服給藥提供可行方案。方法 以離心穩(wěn)定常數(shù)（Ke）為指標(biāo)，采用正交試驗對注射用大豆油的用量、混合乳化劑總量和混合乳化劑的質(zhì)量比進(jìn)行考察，優(yōu)化最佳處方配比?？疾旒{米乳勻機(jī)的運(yùn)轉(zhuǎn)壓力和運(yùn)轉(zhuǎn)次數(shù)對番荔素亞微乳的粒徑大小和分布的影響，優(yōu)化工藝參數(shù)。透射電鏡觀察最佳工藝所得番荔素亞微乳，考察其在生物介質(zhì)中的穩(wěn)定性。采用噻唑藍(lán)（MTT）比色法評價番荔素亞微乳對鼠乳腺癌4T1、人黑色素瘤A875細(xì)胞和人肝癌細(xì)胞HepG2的細(xì)胞毒性。建立鼠源肝癌H22細(xì)胞實體瘤模型，小鼠ig番荔素亞微乳、番荔素油溶液，計算抑瘤率。結(jié)果 番荔素亞微乳的最佳處方配比和制備工藝為：注射用大豆油用量為10%，乳化劑總量為3%，其中精制蛋黃卵磷脂與聚山梨酯80質(zhì)量比為8：2；均質(zhì)壓力為1 500 bar，均質(zhì)次數(shù)為9次。番荔素亞微乳在人工胃腸液中穩(wěn)定，呈球形，粒徑較小，主要在100 nm左右。番荔素亞微乳對4T1、A875、HepG2細(xì)胞IC₅₀值分別為3.082、2.001、1.762μg/mL；ig給藥1 mg/kg番荔素亞微乳與4 mg/kg油溶液對H22荷瘤鼠的抑瘤效果相當(dāng)（65.0% vs 56.5%）。結(jié)論 番荔素亞微乳可以解決番荔素難溶于水、難于給藥的問題，能在不降低療效的同時將用藥劑量降低到傳統(tǒng)油溶液的1/4，在一定程度上具有增效減毒作用。

Objective To prepare annonaceous acetogenins submicroemulsion and study its anti-tumor activities in vitro and in vivo, so as to provide a feasible solution for oral administration of annonaceous acetogenins. Methods Centrifugal stability constant (Ke) was used as indexes, factors of formulation, such as the amount of soybean oil for injection, the total amount of mixed emulsifier, and the mass ratio of mixed emulsifier were optimized by orthogonal test. Effect of process parameters operation pressure and operation frequency on particle size and distribution of annonaceous acetogenins submicroemulsion were investigated. Annonaceous acetogenins submicroemulsion obtained by the best parameters was observed with TEM, and its stability in biological media was also investigated. MTT assay was used to evaluate the in vitro anticancer cytotoxic activity against mice breast cancer 4T1, human melanoma A875, and human hepatic cancer HepG2 cell lines. Solid tumor model on basis of murine hepatic cancer H22 cell were established and mice were ig administrated with annonaceous acetogenins submicroemulsion and oil solution to obtain the tumor inhibition rate. Results The optimal formulation was as following:10% injectable soy oil, 3% combinatorial emulsifier (the weight ratio of purified SPE:Tween 80 being 8:2 in the composition of the combinatorial emulsifier). The optimal homogenization condition was 1 500 bar for 9 cycles. Annonaceous acetogenins submicronemulsion showed good stability in artificial gastric and intestinal juice, and waas spherical, with smaller particle size, mainly around 100 nm. Annonaceous acetogenins submicronemulsion showed strong cytotoxicity against 4T1, AB75, and HepG2 cell lines with IC₅₀ values 3.082, 2.001, and 1.762 μg/mL, respectively. Annonaceous acetogenins submicronemulsion ig administration of 1 mg/kg achieved the same anti-tumor efficacy as that of 4 mg/kg of oil solution on H22 tumor-bearing mice (65.0% vs 56.5%). Conclusion Annonaceous acetogenins submicronemulsion effectively resolves poor solubility and difficulty to be delivered in vivo, and can reduce the dose to 1/4 in contrast to oil solution, and meanwhile maintain similar aiti-tumor efficacy.

國家自然科學(xué)基金資助項目（U1401223，81460734）