基于c-Met和PLK1靶点的新化合物设计、合成及抗肿瘤活性研究

发布时间：2018-03-14 04:48

  本文选题：蛋白激酶　切入点：抗肿瘤　出处：《上海医药工业研究院》2017年博士论文　论文类型：学位论文

【摘要】：c-Met及PLK1激酶抑制剂均是目前新型抗肿瘤药物研究的重要方向。其中,c-Met抑制剂类已有两个药物上市,且多个化合物处于临床研究阶段;PLK1抑制剂多处于临床前和临床研究阶段,暂无药物上市。基于c-Met和PLK1靶点进行相关抑制剂的设计、合成及抗肿瘤活性研究具有较大的学术意义和潜在的应用价值。本文以c-Met和PLK1抑制剂为研究方向,进行新结构类型抑制剂的设计、合成及体外/内活性筛选,以发现具有较好成药性的抗肿瘤活性分子。本论文设计合成了八类共131个新化合物,其中优选化合物SIPI7228和SIPI7067具有较好的体内外活性、且具有毒性小、药代特征良好、潜在心脏毒性小等优点,具有深入研究价值。研究内容及成果如下:1、新化合物设计以c-Met及PLK1激酶抑制剂为研究对象,基于对已上市及临床在研药物基本骨架结构和药效团的分析,依据结构类似性原理采用骨架跃迁、生物电子等排体等有效的药物设计方法进行化合物设计,共设计八个系列(A-D、Ⅰ-Ⅳ)目标化合物。采用计算机辅助药物设计方法分析所设计的新化合物与靶蛋白的结合方式,并根据药理实验结果验证设计思想的合理性,以指导数轮化合物的设计与结构优化。2、新化合物的合成与化学研究合成A-D类c-Met化合物69个,Ⅰ-Ⅳ类PLK1化合物62个,共计131个新化合物,结构均经MS、1H-NMR分析确证。通过八个系列目标化合物的逆合成分析及目标化合物的结构特点,设计了路线较短、方便操作,适合大量制备的合成路线;对于关键中间体的合成方法进行了研究,为样品大量制备奠定了良好基础;同时亦为类似物的合成提供良好的方法借鉴。3、新化合物的抗肿瘤活性研究3.1 c-Met抑制剂的抗肿瘤活性研究A类化合物中SIPI7234和SIPI7235对c-Met的IC50分别为236、275 nmol/L,其中化合物SIPI7234对肿瘤细胞Panc-1、HepG2、Caki-1、HCT116的抗增殖活性与阳性药卡博替尼(Cabozantinib)相当;B类化合物SIPI7100对c-Met的IC50为137 nmol/L,表明氨基嘧啶母核结构对c-Met激酶具有较强的结合力,有利于活性的提高;C类化合物SIPI7224、SIPI7228和SIPI7239的抑酶活性与Cabozantinib相当,抗肿瘤细胞增殖的活性均优于Cabozantinib,确定SIPI7228为优选化合物进行初步成药性评价;D类化合物采用三环结构作为靶酶ATP结合位点,具有一定的创新性,其中化合物SIPI7064、SIPI7067、SIPI7076、SIPI7077均具有较好的抑酶活性及抗肿瘤细胞增殖活性,确定SIPI7067为优选化合物进行初步成药性评价。3.2 PLK1抑制剂的抗肿瘤活性研究Ⅰ类化合物SIPI7473、SIPI7475和SIPI7915的抑酶活性与阳性药Volasertib相当;Ⅱ类化合物SIPI7491对HL-60细胞的抗增殖活性与Volasertib相当,对THP-1细胞的抗增殖活性优于Volasertib,化合物SIPI7489和SIPI7498对THP-1细胞的抗增殖活性与Volasertib相当;Ⅲ类化合物SIPI7482对Hela细胞的抗增殖活性为SBE-13的8倍,化合物SIPI7916对Hela细胞的抗增殖活性为SBE-13的10倍;Ⅳ类化合物抑酶活性不佳,可能是此类化合物保守性较高,改变结构对活性产生较大影响。4、优选化合物SIPI7228和SIPI7067的初步成药性评价对优选化合物SIPI7228和SIPI7067进行相关成药性比较研究,包括急性毒性实验、初步药代实验、hERG钾通道影响实验及裸鼠体内抑瘤实验等。结果表明,化合物SIPI7067和SIPI7228的毒性较小、药代特征良好、潜在心脏毒性小,且表现出较好的体内药效,具有深入研究的价值。综上,本论文所设计的八个系列新化合物丰富了c-Met及PLK1抑制剂的结构类型,及其与相应靶点的构效关系,为高活性化合物的进一步设计奠定了扎实的工作基础,论文研究工作达到预期目的。同时,本文所获化合物设计及合成方面的经验可为该领域创新药物研究提供有益参考。
[Abstract]:C-Met and PLK1 kinase inhibitors are currently an important research direction of new anti-tumor drugs. Among them, c-Met inhibitors have two drugs listed, and a number of compounds in the clinical research stage; PLK1 inhibitors in preclinical and clinical research stage, no drug related design. Inhibitors of c-Met and PLK1 based on the target. Is of great academic significance and potential application value of synthesis and study of antitumor activity. Based on the c-Met and PLK1 inhibitors as the research direction, design new structure type inhibitor screening, synthesis and in vitro / in activity, in order to find good antitumor activity into molecular property. Eight a total of 131 new compounds were synthesized. This paper design, the optimum compound SIPI7228 and SIPI7067 have good activity in vitro and in vivo, and has little toxicity, good pharmacokinetic characteristics, potential advantages of cardiac toxicity etc., With in-depth research value. Research contents and results are as follows: 1. The new compounds are designed based on the c-Met and PLK1 kinase inhibitor as the research object, based on the listed in research and clinical analysis of drug structure and pharmacophore, based on structural similarity principle of the skeleton transition, bioisostere body effective drug design method compound design, design eight series (A-D I - IV) target compounds. Combination of new compounds using analytical methods of computer aided drug design design with the target protein, and according to the rationality of pharmacological experiments validate the design idea, design and structure optimization of.2 guidance in several rounds of synthesis and compounds. Study on the chemical synthesis of A-D c-Met compounds are new compounds 69, I - IV PLK1 compound 62, a total of 131 new compounds. Their structures were confirmed by MS, 1H-NMR analysis confirmed by a series of eight. Inverse synthetic analysis of target compounds and the structural features of the target compound, designed a shorter route, convenient operation, suitable for the preparation of a synthetic route for the synthesis of key intermediates; method was studied for samples prepared to lay a good foundation; at the same time as synthetic analogues provide a good reference for.3. SIPI7234 and SIPI7235 study on antitumor activity of a compound antitumor activity of 3.1 new compounds of c-Met inhibitors of c-Met IC50 were 236275 nmol/L, compound SIPI7234 on tumor cells Panc-1, HepG2, Caki-1, anti proliferative activity of HCT116 and positive drug cabozantinib (Cabozantinib); B compounds SIPI7100 on c-Met the IC50 is 137 nmol/L, showed that the nuclear structure of 2-Aminopyrimidine parent with strong binding force of c-Met kinase, which is helpful to improve the activity of SIPI7224; C compounds, SIPI7228 and SI PI7239 inhibitory activity with Cabozantinib, anti tumor cell proliferation activity was better than Cabozantinib, SIPI7228 was identified as preferred compounds preliminary drugability evaluation; D compounds with tricyclic structure as target enzyme ATP binding sites, with a certain degree of innovation, the compounds SIPI7064, SIPI7067, SIPI7076, SIPI7077 have inhibitory activity and the anti proliferative activity of tumor cells. The SIPI7067 was identified as preferred compounds were preliminary into antitumor activity of compounds SIPI7473 resistance evaluation of.3.2 class I PLK1 inhibitors, SIPI7475 and SIPI7915 inhibitory activity and positive medicine Volasertib; class II compounds SIPI7491 and Volasertib on anti proliferation activity of HL-60 cells, anti proliferative activity than Volasertib in THP-1 cells, SIPI7489 and SIPI7498 compound anti proliferation and Volasertib activity of THP-1 cells is class III; The anti proliferative activity of Hela cells of the compound SIPI7482 is 8 times of SBE-13, the anti proliferative activity of Hela cells was 10 times of SBE-13 SIPI7916 compounds; IV compounds inhibiting enzyme activity of these compounds may be poorly conserved, change the influence of.4 structure on the activity of compounds SIPI7228 and SIPI7067, the preferred initial resistance evaluation a comparative study on the selection of resistance associated into compounds SIPI7228 and SIPI7067, including acute toxicity test, preliminary pharmacokinetic experiment, hERG potassium channel effect and in vivo antitumor experiments. The results show that the less toxic compounds SIPI7067 and SIPI7228, pharmacokinetic characteristics, potential cardiac toxicity, and showed good efficacy in vivo and it has the value of further study. To sum up, the new eight series were designed in this paper enriches the structure of the type c-Met and PLK1 inhibitors, and the corresponding The structure-activity relationship of target points has laid a solid foundation for further design of highly active compounds. The research work is expected to achieve the desired goal. Meanwhile, the experience of compound design and synthesis in this paper can provide a useful reference for the research of innovative drugs in this field.

【学位授予单位】：上海医药工业研究院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R914;R96
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本文编号：1609680