本文選題：蛋白激酶　切入點：抗腫瘤　出處：《上海醫(yī)藥工業(yè)研究院》2017年博士論文　論文類型：學(xué)位論文

【摘要】：c-Met及PLK1激酶抑制劑均是目前新型抗腫瘤藥物研究的重要方向。其中,c-Met抑制劑類已有兩個藥物上市,且多個化合物處于臨床研究階段;PLK1抑制劑多處于臨床前和臨床研究階段,暫無藥物上市。基于c-Met和PLK1靶點進行相關(guān)抑制劑的設(shè)計、合成及抗腫瘤活性研究具有較大的學(xué)術(shù)意義和潛在的應(yīng)用價值。本文以c-Met和PLK1抑制劑為研究方向,進行新結(jié)構(gòu)類型抑制劑的設(shè)計、合成及體外/內(nèi)活性篩選,以發(fā)現(xiàn)具有較好成藥性的抗腫瘤活性分子。本論文設(shè)計合成了八類共131個新化合物,其中優(yōu)選化合物SIPI7228和SIPI7067具有較好的體內(nèi)外活性、且具有毒性小、藥代特征良好、潛在心臟毒性小等優(yōu)點,具有深入研究價值。研究內(nèi)容及成果如下:1、新化合物設(shè)計以c-Met及PLK1激酶抑制劑為研究對象,基于對已上市及臨床在研藥物基本骨架結(jié)構(gòu)和藥效團的分析,依據(jù)結(jié)構(gòu)類似性原理采用骨架躍遷、生物電子等排體等有效的藥物設(shè)計方法進行化合物設(shè)計,共設(shè)計八個系列(A-D、Ⅰ-Ⅳ)目標化合物。采用計算機輔助藥物設(shè)計方法分析所設(shè)計的新化合物與靶蛋白的結(jié)合方式,并根據(jù)藥理實驗結(jié)果驗證設(shè)計思想的合理性,以指導(dǎo)數(shù)輪化合物的設(shè)計與結(jié)構(gòu)優(yōu)化。2、新化合物的合成與化學(xué)研究合成A-D類c-Met化合物69個,Ⅰ-Ⅳ類PLK1化合物62個,共計131個新化合物,結(jié)構(gòu)均經(jīng)MS、1H-NMR分析確證。通過八個系列目標化合物的逆合成分析及目標化合物的結(jié)構(gòu)特點,設(shè)計了路線較短、方便操作,適合大量制備的合成路線;對于關(guān)鍵中間體的合成方法進行了研究,為樣品大量制備奠定了良好基礎(chǔ);同時亦為類似物的合成提供良好的方法借鑒。3、新化合物的抗腫瘤活性研究3.1 c-Met抑制劑的抗腫瘤活性研究A類化合物中SIPI7234和SIPI7235對c-Met的IC50分別為236、275 nmol/L,其中化合物SIPI7234對腫瘤細胞Panc-1、HepG2、Caki-1、HCT116的抗增殖活性與陽性藥卡博替尼(Cabozantinib)相當;B類化合物SIPI7100對c-Met的IC50為137 nmol/L,表明氨基嘧啶母核結(jié)構(gòu)對c-Met激酶具有較強的結(jié)合力,有利于活性的提高;C類化合物SIPI7224、SIPI7228和SIPI7239的抑酶活性與Cabozantinib相當,抗腫瘤細胞增殖的活性均優(yōu)于Cabozantinib,確定SIPI7228為優(yōu)選化合物進行初步成藥性評價;D類化合物采用三環(huán)結(jié)構(gòu)作為靶酶ATP結(jié)合位點,具有一定的創(chuàng)新性,其中化合物SIPI7064、SIPI7067、SIPI7076、SIPI7077均具有較好的抑酶活性及抗腫瘤細胞增殖活性,確定SIPI7067為優(yōu)選化合物進行初步成藥性評價。3.2 PLK1抑制劑的抗腫瘤活性研究Ⅰ類化合物SIPI7473、SIPI7475和SIPI7915的抑酶活性與陽性藥Volasertib相當;Ⅱ類化合物SIPI7491對HL-60細胞的抗增殖活性與Volasertib相當,對THP-1細胞的抗增殖活性優(yōu)于Volasertib,化合物SIPI7489和SIPI7498對THP-1細胞的抗增殖活性與Volasertib相當;Ⅲ類化合物SIPI7482對Hela細胞的抗增殖活性為SBE-13的8倍,化合物SIPI7916對Hela細胞的抗增殖活性為SBE-13的10倍;Ⅳ類化合物抑酶活性不佳,可能是此類化合物保守性較高,改變結(jié)構(gòu)對活性產(chǎn)生較大影響。4、優(yōu)選化合物SIPI7228和SIPI7067的初步成藥性評價對優(yōu)選化合物SIPI7228和SIPI7067進行相關(guān)成藥性比較研究,包括急性毒性實驗、初步藥代實驗、hERG鉀通道影響實驗及裸鼠體內(nèi)抑瘤實驗等。結(jié)果表明,化合物SIPI7067和SIPI7228的毒性較小、藥代特征良好、潛在心臟毒性小,且表現(xiàn)出較好的體內(nèi)藥效,具有深入研究的價值。綜上,本論文所設(shè)計的八個系列新化合物豐富了c-Met及PLK1抑制劑的結(jié)構(gòu)類型,及其與相應(yīng)靶點的構(gòu)效關(guān)系,為高活性化合物的進一步設(shè)計奠定了扎實的工作基礎(chǔ),論文研究工作達到預(yù)期目的。同時,本文所獲化合物設(shè)計及合成方面的經(jīng)驗可為該領(lǐng)域創(chuàng)新藥物研究提供有益參考。
[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.

【學(xué)位授予單位】：上海醫(yī)藥工業(yè)研究院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：R914;R96
，

本文編號：1609680