【作者】 王静丽；

【导师】 赵临襄；

【作者基本信息】 沈阳药科大学 ， 药物化学， 2004， 博士

【摘要】 对天然抗肿瘤萜类化合物进行系统的构效关系研究，发现α-亚甲基-γ-丁内酯或α-亚甲基环戊酮是绝大多数倍半萜类或二萜类化合物具有抗肿瘤活性的必需结构。计志忠教授等将这些天然结构简化，设计合成了以WB852为代表的近五百个2-取代胺甲基-5-取代亚甲基环戊（己）酮类化合物，这些化合物具有抗癌活性显著、毒性低、结构简单和易合成等特点。初步作用机理研究结果表明，WB852可经消除反应，生成α，β-不饱和酮结构；然后与还原型谷胱甘肽（GSH）结合，生成1，4-Micheal加成产物。WB852通过降低细胞内GSH浓度，抑制谷胱甘肽转移酶π（GSTπ）的活性(IC₅₀为30μmol／L)，诱导细胞凋亡。 为深入研究2-取代胺甲基-5-取代亚甲基环戊（己）酮类化合物的抗癌作用机理，寻找新型肿瘤细胞凋亡诱导剂，本文在总结这类化合物的抗癌活性、构效关系的基础上，以WB852为先导化合物，对取代亚甲基、取代胺甲基、羰基部分进行系统的结构修饰和改造，设计和合成了三类共79个WB852类似物，其中有52个2-取代胺甲基-5（6）-取代亚甲基环戊（己）酮类化合物（W1-1～W1-21，W2-1～W2-11，W3-1～W3-5，W4-1～W4-15），11个2-（取代）亚甲基-5-取代芳亚甲基环戊酮类化合物（J1-1～J1-9，J2-1～J2-2），16个2-（E）-取代苯亚甲基-5（6）-取代胺甲基-1-取代苯基环戊（己）醇类化合物（L1-1～L1-16）。经IR、¹H-NMR和MS等分析数据确证其中72个为未见文献报道的新化合物。在合成中间体中，有17个化合物未见文献报道，均经¹H-NMR证实。 首次将Eschenmoser’s salt用于2-胺甲基-5-取代亚甲基环戊酮类化合物的制备，简化了操作过程，避免了经典的Mannich反应常见的聚合、脱胺基化等副反应，大大提高了反应收率（由低于20％提高到接近60％）。通过控制反应温度，提高了反应的区域选择性，成功地制备了2-取代胺甲基-5-取代甲基环戊酮类化合物（W2-9～W2-10）。 总结了目标化合物的IR规律与EI／MS谱规律；根据含取代苯丙烯酸酯结构化合物（W4-1～W4-15）双键两氢原子间的偶合常数J=15.9Hz，判断双键上两氢原子为反式构型；利用¹H-¹HCOSY谱配合局部放大图，确定了化合物2-二苯甲基-5-二甲基胺甲基环戊酮盐酸盐（W2-9）、2-（E）-[4-[3-（4’-氯代苯基）丙烯酰氧基]]-苯亚甲基-5-二甲胺基甲基更多还原

【Abstract】 A large number of natural diterpenes and sesquiterpenes have remarkable bioactivity. The presence of α-methylene cyclopentanone appearing in diterpenes and α-methylene-γ-lactones appearing in sesquiterpenes plays a significant r ole for anti-tumour and cytotoxic activities. Inspired by the anti-tumour effect of these natural components we designed and synthesized a series of 2-alkylaminomethyl-5-(E)-alkylidene cyclopentanone hydrochlorides. WB852, one of the representatives of such compounds, had significant anti-tumor activities in vitro and in vivo and was an effective apoptosis-inducing agent in leukemia cells and it decreased intracellular reduced glutathione (GSH) of leukemia cells. The mass spectrum of a GSH adduct of WB852 indicated that the mechanism of action for WB852 and its analogues may proceed via an elimination-Michael addition process to form an α-methylene group, followed by a 1,4-Michael addition with free cysteine sulfhydryl groups including GSH to form thiol-addition product. GSH plays an important role in cells to prevent cell death as a substrate of glutathione-s-transferases (GST) which involve in the drug resistance. Among the GST family it has been found that GSTπ was increased in chemotherapy-resistance cancer cells and thus GSTπ could be used as a potential target for the cancer treatment. The potential GSH binding ability of WB852 suggests that it may inhibit GSTπ activity directly or indirectly, and would be potent anticancer agents in tumor cells even with higher GSTπ activity. WB852 as a promising lead compound was supposed to induce tumor cells apoptosis through binding cellular GSH, inhibiting GSTπ activity, decreasing intracellular GSH and generation of oxidative stress.Based on the structure-activity relationships (SAR) of antitumor activities of cyclopentanone Mannich base hydrochlorides summarized previously, with WB852 as a lead compound, modified systemically from the following three aspects of WB852: alkylaminomethyl, alkylidene and ketone group for further study of the SAR, three types of更多还原