靶向肝癌干细胞及多药耐药逆转药物异紫堇碱类似物的制备、构效关系及作用机制研究

Tumor recurrence, metastasis and multiple drug resistance were the main factors for the failure of tumor therapy and the cancer stem cells (CSCs) is the main decisive roles for those factors. The investigation about the multiple targeted drugs based on CSCs, special tumor cells target or the reversion of multiple drug resistance were the hot spot in the chemtherapy research. Studies showed the aporphine alkaloids have more biological function, which could be used as a multiple targeted drug selectively actived on tumor cell epidermal growth factor receptor (EGFR), CSCs of hepatoma carcinoma cell (HCC) and reversed the multiple drug resistance. This application based on the isocorydine which is the main anticancer chemical constituent of the northwest characteristic plant resources of Dicranostigma leptopodum (Maxim) Fedde, through extraction and separation, the chemical synthesis and the structural modification, the isocorydine derivatives library will be prepared, the molecular docking study on the targeted EGFR and the research of 3D - QSAR will be explored to obtain the structure-activity relationship and pharmacophore of this aporphine alkaloids. According to the characteristic of the targeted drugs and the reversion of multiple drug resistance special, rational drug design will be applied to optimize the target molecular structure. The molecule targets and the signal transduction pathways will be illuminated through the investigation of pharmacological mechanism of those derivatives. Through those investigation, the applied project will provide the scientific basis and the beneficial exploration for the research of newly multiple targeted and the reversion of multiple drug resistance.

肿瘤的复发、转移和多药耐药是肿瘤治疗失败的关键因素，肿瘤干细胞在复发、转移和多药耐药的产生中具有决定性作用，研究基于肿瘤干细胞及特殊肿瘤细胞靶标的靶向药物和多药耐药逆转的多靶点抗癌药物是目前抗癌药物研究的热点，研究显示阿普菲类生物碱可以作为多靶点药物选择性作用于肝癌细胞表皮细胞生长因子受体、肝癌细干细胞以及具有多药耐药逆转等生物学功能。本项目以西北特色植物秃疮花中的抗癌主要活性成分异紫堇碱为先导，通过提取分离、化学合成、结构改造制备异紫堇碱类似物化合物库、针对EGFR的分子对接以及抗癌活性3D-QSAR研究，探讨该类化合物的构效关系和药效团，并针对靶向药物和多药耐药逆转特性进行合理药物设计，优化该类化合物目标分子结构；通过体内外作用机理研究明确目标分子作用靶点和信号转导通路，为新的高效多靶点及多药耐药有效逆转的靶向肝癌干细胞药物研究提供科学依据。

基于西北特色植物资源深度开发，以秃疮花和云南地不容为药材资源，采用天然产物化学手段分离制备大量的异喹啉类生物碱类化合物；并以高含量活性成分异可利定为先导物，采用有机合成手段，通过化学结构衍生制备得到大量的异紫堇碱类化合物；通过抗癌活性筛选得到目标化合物NICD/FICD/COM33；通过目标化合物的抗癌活性及其机制研究发现异可利定衍生物的药理作用机制及其靶点：NICD具有选择性作用于肝癌细胞的IGF2BP3和CD133+为标记的肿瘤干细胞，从而引起ABCG2和ABCB1耐药相关蛋白的下调，能够逆转肝癌细胞的耐药性，同时NICD具有索那菲尼的协同增效作用；NICD可以靶向作用于转录因子C/EBP，从而引起GADD45A和P21的上调，诱导肝癌细胞凋亡和肝癌细胞G2周期阻止；NICD可以通过靶向转录因子E2F1引起整合素(ITGA1)的下调而阻止肝癌细胞的迁移和侵袭。. 本项目前期通过异紫堇碱衍生物的靶向干预，建立了肿瘤干细胞参与的肿瘤多药耐药逆转、肝癌细胞凋亡以及肿瘤转移侵袭三者之间的相互关系，使一种药物解决三个肝癌治疗中的关键科学问题成为可能。通过C-7氨基引入经典靶向药物的高效抗癌基团使得异紫堇碱类化合物的抗癌活性显著提高，并筛选得到目标化合物COM33正在进行抗肝癌一类新药研发。. 得到异紫堇碱类（异喹啉类）生物碱化学结构衍生的启发，申请人又探索性发展一类异吲哚啉酮类生物碱的合成方法，并巧妙的将资源型特色显著的当归药材的药效标志物藁本内酯通过环丙胺类仲胺基团的引入构建了异吲哚啉酮类生物碱结构骨架。通过药理活性筛选发现该类化合物虽然抗癌活性有限，但具有血小板聚集抑制作用，可以促进毛细血管新生和侧支循环重构，具有明确抗脑卒中活性，目前正在开展抗脑卒中一类创新药物开发。