【作者】 解丽娟；

【导师】 钱旭红；

【作者基本信息】 大连理工大学 ， 应用化学， 2009， 博士

【摘要】 根据以DNA为靶点抗肿瘤药物的作用机理,通过对天然产物呋喃香豆素、抗癌先导化合物amonafide和苊并吡咯杂环类化合物进行结构改造,本文设计、合成、表征了三系列以DNA为靶点的抗肿瘤化合物:1.以具有良好DNA嵌入功能的光化治疗试剂呋哺喹啉酮为母体,通过在其喹啉酮环氮原子上引入柔性侧链,设计了一系列N-取代的角型呋哺喹啉酮衍生物。以间苯二胺为原料,经佩希曼缩合、重氮化、水解、取代、环合、偶联六步反应合成了十二个目标化合物（其中十一个未见报导）。体外抗肿瘤活性测试显示,碱性氨基侧链的引入有助于改善母体的抗肿瘤功效,其中活性最好的化合物1h（N-（2-dimethyl amino-ethyl）-2-（4,8,9-trimethyl-2-oxo-2H-furo-[2,3-h]quinolin-1-yl）-acetamide）对P388和A549肿瘤细胞的IC₅₀值分别为14.45μM和20.54μM,同样测试条件下母体化合物的IC₅₀值分别为＞100μM和99.29μM。2.为了即保留萘酰亚胺类抗癌先导amonafide的生物活性,又降低其副作用,设计了5-位脂肪氨基和5-位芳香基团取代的萘酰亚胺衍生物。以1,8-萘酐为原料,经溴化、胺化和偶联三步反应合成目标化合物。在最后一步反应中,由于萘酰亚胺结构的特点,导致其5-位用传统的化学方法难以发生亲核取代反应。本论文首次应用碘化亚铜/脯氨酸和四三苯基膦钯催化的Suzuki偶联反应,合成八个新的5-位脂肪氨基取代的amonafide类似物和五个5-位芳香基团取代的萘酰亚胺衍生物（四个未见报导）。通过亲核取代反应得到了相应的八个6-位脂肪氨基取代的萘酰亚胺衍生物。这些新的萘酰亚胺化合物中有八个对HeLa和P388D1肿瘤细胞显示出优于amonafide(IC₅₀值分别为6.02μM和0.68μM)的体外抗肿瘤活性,对HeLa细胞3a(5-（dimethylamino-ethylamino）-2-[2-（dimethylamino）ethyl]-1H-benzo[de]-isoquinoline-1,3（2H）-dione,IC₅₀=0.69μM)、3b(5-（dimethylamino-propylamino）-2-[2-（dimethylamino）ethyl]-1H-benzo[de]isoquinoline-1,3（2H）-dione,IC₅₀=0.71μM)、4b(6-（dimethylamino-propylamino）-2-[2-（dimethylamino）ethyl]-1H-benzo[de]isoquinoline-1,3（2H）-dione,IC₅₀=0.64μM);对P388D1细胞3b(IC₅₀=0.23μM)、4b(IC₅₀=0.23μM)。由于结构中没有伯胺基团,可以避免类似amonafide的副作用。对比新合成的5-位氨基取代物和相对应的6-位氨基取代物体外抗肿瘤活性,发现6-位取代物的体外抗肿瘤活性普遍好于5-位取代物。这项研究不仅得到新的具有抗肿瘤活性的化合物,扩大临床药物筛选范围,而且丰富了萘酰亚胺类抗肿瘤化合物构效关系的研究。3.设计、合成了新的苊并吡咯羧酸类衍生物。以苊醌为原料,经KnoevenagelCondensation,环构化、水解、芳香亲核氢取代四步反应合成九个目标化合物（未见报导）。其中5d（3-（3-dimethylamino-propylamino）-8-oxo-8H-acenaphtho[1,2-b]pyrrole-9-carboxylicacid）对HL-60细胞的IC₅₀为13.1μM,5g（3-Piperidin-8-oxo-8H-acenaphtho[1,2-b]-pyrrole-9-carboxylic acid）对HeLa、HL-60、HCT-8、A-3375和MCF-7细胞的IC₅₀分别为20.6、20.5、24.4、18.4和14.5μM,具有较好的体外抗肿瘤活性。流势细胞术法检测发现对HeLa细胞,化合物5d可以引起细胞周期阻滞于S期和诱导凋亡,同样实验条件下5g诱导凋亡:光谱学方法和粘度测试显示这两个化合物以同样的方式与DNA结合。4.利用紫外、荧光、圆二色谱及粘度测试方法研究了上述化合物与DNA之间的相互作用,结果显示这些化合物主要是通过嵌入结合的方式与DNA相互作用的。嵌入常数均达到10⁵数量级,是中等强度DNA嵌入剂。更多还原

【Abstract】 Based on the action mechanism of antitumor agents that target DNA,three series of compounds were designed,synthesized and charactered by structural modify to natural product furocoumarin and acenaphtho-pyrrole heteroaromatic system and the derivative of lead amonafide.The following studies are made.1.Based on parent compound,furoquinolinone,which is a phototherapeutical agent, N-substituted angular furoquinolinone derivatives were designed by introducing of the side chain to the N-position of furoquinolinone.Twelve target compounds（eleven compounds are not reported） were prepared from m-phenylenediamine and ethyl acetoacetate by six steps including Pechmann condensation,diazotization,hydrolysis,substitution,cyclization and coupling reaction.The evaluation for antitumor activities in vitro indicated that the introduction of amino side chain is an effective way to improve the antitumor activity of the parent.Compound 1h N-（2-dimethylamino-ethyl）-2-（4,8,9- trimethyl-2-oxo-2H- furo[2,3-h]-quinolin-1-yl）-acetamide exhibited the highest activities against P388 and A549 cell lines and its IC₅₀ values are 14.45μM and 20.54μM,respectively.However,the IC₅₀ of the parent compound are＞100μM and 99.29μM at the same experiment,respectively.2.To retain the biological activity of the amonafide and reduce the side effects,5-position aliphatic amine substituted naphthalimide and 5-position aromatic substituted naphthalimide were designed.The target compounds were synthesized from naphthalic anhydride by three steps including bromization,amination and coupling reaction.The nucleophilic substitution reaction at the 5-position of the naphtahlimide is difficult to occur by traditional chemical method due to electron-charge accumulation induced by strong electron-withdrawing ability of carbonyl.Therefore,the third step reaction is the most difficult.In this paper,we first reported the synthesis of 5-aliphatic amine substituted and 5-aromatic substituted naphthalimide derivatives via CuI/proline catalyzed coupling reaction and Suzuki coupling reaction,respectively.Eight 6-aliphatic amine substituted naphthalimide derivatives were synthesized via nucleophilic substitution reactions.The eight new compounds exhibited better activity than amonafide(the IC₅₀ vaIues are 6.02μM and 0.68μM,respectively)against HeLa and P388D1 cell under the same experimental conditions.Among the compounds,3a (5-（Dimethylamino-ethylamino）-2-[2-（dimethylamino）ethyl]-1H- benzo[de]- isoquinoline-1,3 （2H）-dione,IC₅₀ = 0.69μM),3b(5-（Dimethylaminopropylamino）- 2-[2-（dimethylamino） ethyl]- 1H-benzo[de]isoquinoline- 1,3（2H）-dione,IC₅₀ = 0.71μM),4b(6-（dimethylaminoproplamino） -2-[2-（dimethylamino）ethyl]-1H-benzo[de]isoquinoline-1,3（2H）-dione,IC₅₀ = 0.64μM);against P388D1,3b(IC₅₀ = 0.23μM),4b(IC₅₀= 0.23μM).The compounds can avoid the side effect of amonafide due to lack of a primary amine in their structure.The antitumor activities in vitro of the 6-amino substituted naphthalimide derivatives are better than those of 5-amino substituted.Therefore,this work not only can obtain new compounds with antitumor activities for clinical screen,but also expand the structure activity relationships studies of the naphthilimides.3.New 8-oxo-SH-acenaphtho[1,2-b]pyrrole-9-carboxylic acid derivatives were designed and synthesized.Nine target compounds were synthesized from acenaphthene quinone by four steps including Knoevenagel condensation,cyclization,hydrolysis and S_NAr^H reaction. Among the compounds,5d（3-（3-Dimethylamino-propylamino）-8-oxo-8H-acenaphtho[1,2-b]pyrrole-9-carboxylic acid） and 5g（3-Piperidin-8-oxo-8H-acenaphtho[1,2-b]pyrrole-9-carboxylic acid） show better antitumor activities in vitro,the IC₅₀ value of 5d against HL-60 cell is 13.11μM and the IC₅₀ of 5g against HeLa,HL-60,HCT-8,A-3375 and MCF-7 cell are 20.61,20.52,24.42,18.43 and 14.51μM,respectively.Cell cycle analysis indicated that 5d could induce S-phase arrest followed by apoptosis and 5g only induced apoptosis,although the both compounds could bind to DNA in same way.4.DNA-binding of the above mentioned compounds are investigated by fluorescent, UV-vis,CD spectroscopic techniques and viscosity measurement.The results indicate that the compounds bind to DNA mainly via intercalation.The Scatchard-binding constants in 10⁵ order of magnitude are actually moderate DNA-intercalators.更多还原