不饱和漆酚抗肿瘤多配体靶向聚合体胶束合成与载药控释机理研究

Raw lacquer is a kind of important forest products in China, its main active constituents of urushiol possess prominent antitumor biological activity. However, as urushiol can cause hypersensitive reaction, it is sensitive to oxidation and polymerization, and the separation of unsaturated urushiol (USU) monomers are difficult, so the application of urushiol in drug development will be limited. In this study the refined USU in raw lacquer will be used as raw material, we will study acetal protection and D-A chemical modification mechanism of active groups of urushiol, and separate catechol USU acrylic ester by medium pressure column and countercurrent chromatography. We will design and synthesize pH- responsed USU-PBAE-PEG-Gal amphiphilic polymers, by the amidation and deprotection of USU acrylic ester , with galactose (Gal) as targeting ligands, and by regulating the proportion of hydrophobic/hydrophilic balance via changing the molecular weight of poly amino ester (beta) (PBAE) and polyethylene glycol (PEG). Moreover, urushiol amphiphilic polymer micelle will be prepared with the amphiphilic targeting ligands of TLS11a-C18 and CendR-C18. Meanewhile, the synthesis mechanism and structure characteristics of amphiphilic polymer micelle will be studied, the in vitro slow control mechanism of urushiol amphiphilic polymer micelle loaded with anti-cancer drugs of DOX or paclitaxel will be clarified, and its drug loading capacity and release performance will be evaluated. We will comparatively analysize in vivo distribution performance and in vivo antitumor effects of different targeting drug system of micelle, in order to clarify mechanism of antitumor attenuated synergies of unsaturated alkyl phenol in raw lacquer combined with chemotherapeutic drugs, which will provides the basis for the antitumor drug development of urushiol.

生漆是我国重要的林产品,生漆中漆酚具有抗肿瘤生物活性。由于漆酚易过敏、氧化和聚合,不饱和漆酚(USU)单体分离困难,限制漆酚药物开发应用。本研究以精制USU为原料，研究漆酚活性基团缩醛保护和D-A化学修饰机理,通过中压柱和逆流色谱分离邻苯二酚型USU丙烯酸酯。研究USU丙烯酸酯酰胺化和脱保护反应,以半乳糖(Gal)为靶向配体,通过改变PBAE和PEG分子量,调控疏水/亲水平衡比例,设计合成pH-响应USU-PBAE-PEG-Gal两亲聚合物,并通过两亲靶向配体TLS11a-C18和CendR-C18制备漆酚两亲聚合物胶束,研究其合成机理和结构特征,阐明漆酚两亲聚合物胶束负载DOX或紫杉醇等抗癌药体外药物缓控机理, 评价其药物负载能力和释放性能。对比分析不同胶束靶向载药体系的体内分布性能和体内抗肿瘤效果,阐明生漆不饱和烷基酚联合化疗药抗肿瘤减毒增效的机理,为漆酚抗肿瘤药物开发提供基础。

生漆是传统的天然涂料，漆酚作为生漆的主体化合物，大量研究主要集中在漆酚自聚合、金属聚合及漆酚有机聚合物的耐化学性能和力学性能等功能材料领域。本项目重点从生漆中分离纯化C15三烯漆酚，再通过化学合成得到了C15三烯漆酚Pechmann、D-A、卤素、硫醇长链烷烃、苯硼酸、氨基亚砜和三氮唑等漆酚衍生物及漆酚基大分子载药胶束材料，并对其抗肿瘤生物活性和安全性等进行评价，为高附加值漆酚衍生物及漆酚基医药胶束材料的药物开发提供理论和应用基础。.（1）从生漆中分离纯化得到C15单烯漆酚（2a）和C15三烯漆酚(2b)，合成得到C15三烯漆酚Pechmann型衍生物(2c)，均有较好的抑制HepG2细胞增殖的活性（2c＞2a＞2b），半数抑制浓度(IC50)分别为29.3 μM、55.5 μM和27.1 μM。当漆酚Pechmann型化合物2c与紫杉醇的比例为6.775 μM：7.9 ng/mL时，协同作用最强，协同指数（CI）达到0.641。.（2）基于Diels-Alder的[4+2]反应，将马来酸酐五元环骨架和不同电负性的-NH，-O及-F，-Cl，-Br卤素引入漆酚烯烃侧链，合成了8个新型漆酚基D-A型衍生物3e~3l，化合物3e和3k对MKN-45和HCT-116的抑制效果最好，分别为3.2 μM、2.6 μM对MKN-45和1.2 μM、0.66 μM对HCT-116。.（3）化合物4a、4d和4i分别为2位酚羟基醚化引入了巯基、苯硼酸和氨基亚砜的产物；它们对抑制HepG2增殖的IC50值提高到7~15 μM，而对正常肝细胞LO2的IC50减弱到120~150 μM；其中，漆酚含巯基衍生物4a对HepG2和LO2的IC50分别为7.886 μM和150 μM。 .（4）具有富电子结构的漆酚三氮唑类化合物5d中三氮唑作为常用药效团，对HepG2具有较高的抗肿瘤活性，IC50值为2.833 μM。而漆酚Pechmann化合物5j对LO2细胞毒性(IC50=198 μM)低于漆酚三氮唑类化合物5d，对LO2的IC50值比HepG2高出20倍以上，对正常肝细胞的LO2有较低毒性。.（5）通过Michael加成反应，制备了一种紫杉醇和漆酚协同释放的pH靶向漆酚基胶束BPAU-NH2-Gal-PTX，并表征了其化学结构。BPAU-NH2和BPAU-NH2-Gal包埋紫杉醇胶束的粒