Preparation and the function of a novel surfactant vesicle with a recognition ability of the specific antigen of cancer cell toward practical clinical application

本工作的主要目的是将用于癌症治疗的新型囊泡在药物释放系统(DDS)中的实际应用，制备一种非离子表面活性物质囊泡，该囊泡由山梨醇单油酸酯组成。作为肿瘤靶向导弹装置的固定化配体，我们选择了两种配体：(1)新型藻类凝集素ESA(Eucheuma Serra Aggaginin：)和(2)抗肿瘤细胞抗原的单抗。结果表明，Span 80是山梨醇酯单酯、双酯和四酯异构体的表面活性混合物，囊泡膜是通过热力学自组装形成恒定的异构体。主要成分为双酯(约60%)。与常用的天然磷脂脂质体相比，SPAN80囊泡具有更高的流动性和更好的膜融合性、耐热性、代谢性和低成本等特性。体内实验表明，ESA固定的SPAN80囊泡(ESA囊泡)能与结肠腺癌(Colo201，Colon26)等肿瘤细胞发生特异性结合。我们还可以揭示，注射到患有结肠腺癌的小鼠体内的ESA囊泡，通过血壁转移并靶向肿瘤，随后肿瘤发生凋亡死亡(隧道分析证实)。肝内蓄积的囊泡在48小时内完全代谢。含有抗癌药物多西紫杉醇(TXT)的SPAN80囊泡对小鼠肿瘤有很好的抑制作用。未观察到囊泡大小和副作用的影响。基因组分析还阐明了该细胞的凋亡机制(通过Caspae3和8)。我们还可以通过注射固定了抗EGFR抗体的免疫小室来证明有效地控制了模型人脑肿瘤(来自ERM5-1细胞)的生长。此外，以免疫小球为载体，成功地对ERM5-1肿瘤进行了体内基因转导。

The main purpose of this work is the practical clinical application of the novel vesicle for a cancer therapy in drug delivery system (DDS), preparing a non-ionic surfactant vesicle, composed of Span80 (sorbitan monooleate). As the immobilized ligand as a "tumor-targeting missile device" on the Span80 vesicle, two species of ligands were chosen: (1) novel alga lectin ESA (Eucheuma Serra agglutinin:) and (2) monochronal antibody against the antigen of a tumor cell.We revealed that Span80 was a surfactant mixture of sorbitan ester isomers of mono-, di-tri-and tetra-esters and the vesicle membrane was constructed as constant composition of the isomers by a thermodynamic self-assembly. The main component was diester (about 60%). Span80 vesicle had higher fluidity and superior characteristics such as membrane fusion, heat-satiability, metabolism and low-cost, compared with usual liposome with natural phospholipids. The ESA-immobilized Span80-vesicle (ESA vesicle) was revealed to specifically combine to cancer cells, such as colon adenocarcinoma (Colo201, Colon26) in vivo. We could also reveal that the ESA vesicle, injected to a mouse with the colon adenocarcinomas, transferred through the blood wall and targeted at the tumor following by the apoptotic death (confirmed by TUNNEL analysis) of the tumor. The vesicle accumulated in the liver was perfectly metabolized within 48 hours. The Span80 vesicle containing anticancer drug of docetaxcel (TXT) could perfectly controlled the mouse tumor. The influence of the vesicle size and the side effect could not be observed. The apoptotic mechanism (through Caspae3 and 8) was also clarified by genome-analysis. We could also demonstrate the effective control of the growth of the model human brain-tumor (from ERM5-1 cell) by injecting the immunovesicle immobilized with anti-EGFR antibody. Moreover, using the immunovesicle as a vector, gene transfection to the ERM5-1 tumor in vivo was successfully performed.