蛋白激酶B抑制剂的设计合成、生物活性测定、分子模拟研究及Biginelli反应研究

发布时间：2018-03-05 06:25

本文选题：ATP竞争性抑制剂　切入点：变构抑制剂　出处：《浙江大学》2014年博士论文　论文类型：学位论文

【摘要】：蛋白激酶B (PKB/Akt)作为良好的抗肿瘤药物靶点,已有多种PKB抑制剂被广泛研究并应用于肿瘤的治疗。其中ATP竞争性抑制,变构抑制剂,非ATP竞争性抑制剂和假底物抑制剂是当前研究最广泛的PKB抑制剂。但是由于PKB亚型之间及PKB与其它激酶间的氨基酸序列存在高度同源性,因此PKB抑制剂的选择性设计是PKB抑制剂研究领域的一个棘手的难题。蛋白激酶选择性抑制机理的研究可以为选择性抑制剂的设计提供良好的基础。而分子动力学模拟方法由于可以通过评价蛋白激酶-抑制剂复合物体系的稳定性以及蛋白激酶与抑制剂之间的亲和力来研究蛋白激酶选择性抑制的机理,并在蛋白激酶选择性抑制剂研究方面得到了广泛的应用。而二氢嘧啶酮类化合物和螺杂环取代的吲哚类化合物分别具有广泛的生物活性与药理活性,可以作为抗肿瘤药,抗微生物药等。因此通过Biginelli反应合成二氢嘧啶酮类化合物和螺杂环取代的吲哚类化合物引起了人们广泛的研究兴趣。因此,本论文围绕着PKB抑制剂设计合成、生物活性测试、分子模拟研究和Biginelli反应进行了以下研究： (1)、本文通过对PKA, PKBa, PKBa计算机模拟突变体和吡咯并嘧啶类抑制剂的9个复合物体系进行分子动力学模拟研究,在分析吡咯并嘧啶类抑制剂结合模式的基础上,从模拟的角度合理地解释了PKB抑制剂相对于PKA选择性抑制PKB的机制,并成功地验证了已报道的关于于PKB抑制剂相对于PKA选择性的一些实验研究结果。同时也提出了这类抑制剂的结构修饰规律,指导这类抑制剂的设计。 (2)、本文通过对PKBa, ROCK1, PKBa计算机模拟突变体和吡啶类抑制剂的7个复合物体系进行分子动力学模拟研究,在分析吡啶类抑制剂结合模式的基础上,从模拟的角度合理地解释了PKB抑制剂相对于ROCK1选择性抑制PKB的机制,成功地验证了已报道的关于PKB抑制剂相对于ROCK1选择性的一些实验研究结果。同时也提出了这类抑制剂结构修饰的规律,指导这类抑制剂的设计。 (3)、本文通过对PKBa与变构抑制剂的3个复合物体系进行分子动力学模拟研究,从模拟的角度合理地解释了变构抑制剂的作用机制和结合模式,成功地验证了已报道的关于PKB变构抑制剂的实验研究结果,并提出了这类抑制剂结构修饰的规律,指导这类抑制剂的设计。 (4)、本文通过对PKB非ATP竞争性抑制剂进行3D-QSAR研究,同时对构建的5个复合物体系进行分子动力学模拟研究,从模拟的角度合理地解释了非ATP竞争性抑制剂的结合模式和PKB亚型选择性机制,成功地验证了已报道的关于非ATP竞争性抑制剂的实验研究结果,同时根据非ATP竞争性抑制剂的3D-QSAR研究结果,提出了非ATP竞争性抑制剂结构修饰的规律,指导这类抑制剂的设计。 (5)、本文通过PKBa与肽类假底物抑制剂的3个复合物体系进行分子动力学模拟研究,从模拟的角度合理地解释了肽类假底物抑制剂的结合模式,成功地验证了已报道的关于PKB假底物抑制剂的一些实验研究结果。同时也提出了假底物抑制剂结构修饰的规律,指导这类抑制剂的设计。 (6)、结合PKB分子模拟的结果及相关实验结果,本文主要设计了9类化合物和合成了89个化合物,并对其中51个化合物进行了体外PKBa激酶的抑制实验。其它化合物有待进行激酶抑制实验。 (7)、本文研究了三氟甲磺酸镥崔花的和氯甲基二甲基氯硅烷(CMDMCS)参与的Biginelli反应。反应以醛/靛红化合物/缩醛,1,3-二羰基化合物/缩酮,脲/硫脲为起始原料,反应高收率地得到了22个二氢嘧啶酮类化合物。其中,三氟甲磺酸镥可回收和重复使用,而氯甲基二甲基氯硅烷廉价易得,因而为该类化合物的制备提供了新的合成途径,同时为该类化合物的进一步相关研究奠定了基础。
[Abstract]:Protein kinase B (PKB/Akt) as anti-tumor drug targets is good, there are many kinds of PKB inhibitors are widely studied and applied in the treatment of cancer. The ATP competitive inhibitor, allosteric inhibitors, non competitive inhibitors of ATP and the false inhibitor is current research most widely used PKB inhibitors. But due to the amino acid sequence between PKB subtypes and PKB and other kinases exist between highly homologous, so the design of selective PKB inhibitors is a difficult problem in the research field of PKB inhibitors.
Study on protein kinase selective inhibition mechanism can provide a good foundation for the design of selective inhibitors. The molecular dynamics simulation method for stability evaluation mechanism can be through protein kinase inhibitor complex system and protein kinase inhibitors and inhibition of protein kinase selective affinity to study, and has been widely used in the study of protein kinase selective inhibitors.
Indole compounds and two hydrogen pyrimidinone compounds and spiro heterocyclic substituted respectively with biological activity and extensive pharmacological activity, can be used as antitumor drugs, anti microbial drugs. Therefore indole compounds synthesized through the Biginelli reaction of two hydrogen pyrimidinone compounds and substituted spiro heterocyclic compounds have attracted considerable research interests.
Therefore, this paper focuses on the design and synthesis of PKB inhibitors, biological activity test, molecular simulation research and Biginelli reaction.
(1), based on PKA, PKBa, 9 of the complex system simulation of mutant and pyrrolopyrimidine inhibitor PKBa by computer molecular dynamics simulation research, in the analysis of pyrrolopyrimidine inhibitor binding on the basis of the model, from the simulation can explain the mechanism of PKA PKB inhibitors with respect to the selective inhibition of PKB. And successfully verified reported about PKB inhibitors with respect to some experimental results on the selectivity of PKA. At the same time also proposed structure modification law of such inhibitors, guiding the design of such inhibitors.
(2), based on PKBa, ROCK1, 7 complex system simulation mutants and pyridine inhibitor PKBa by computer molecular dynamics simulation study, based on the analysis of pyridine inhibitor binding mode, from the simulation angle reasonably explained the mechanism of ROCK1 PKB inhibitors for the selective inhibition of PKB, successfully verified it has been reported about PKB inhibitors with respect to some experimental results on the selectivity of ROCK1. At the same time put forward the modified inhibitors of this kind of guiding the design of inhibitors.
(3), the molecular dynamics simulation research through 3 complexes of PKBa and allosteric inhibitors, from the simulation angle reasonably explained the mechanism of allosteric inhibitors and binding mode, successfully verified about the PKB allosteric inhibitor of the experiment research results, and put forward the modification of these inhibitors the structure of law, guiding the design of such inhibitors.
(4), based on the study of 3D-QSAR PKB non competitive inhibitors of ATP, while the 5 complex system of molecular dynamics simulation, from the simulation can explain the non competitive inhibitors of ATP binding mode and PKB subtype selective mechanism to verify the experimental results about non competitive inhibitors of ATP have been reported, according to the results of 3D-QSAR study on non competitive inhibitors of ATP, proposed the modified non ATP competitive inhibitors of this kind of guiding the design of inhibitors.
(5), this paper studied by molecular dynamics simulations of 3 complexes with PKBa peptide pseudosubstrate inhibitor, from the simulation angle can reasonably explain the binding mode of peptide pseudosubstrate inhibitor, has been successfully verified reports about PKB inhibitors pseudosubstrate some experimental results. At the same time also proposed modified materials inhibitor of the design direction of this inhibitor.
(6) combined with the results of PKB molecular simulation and related experimental results, 9 compounds and 89 compounds were synthesized, and 51 compounds were tested for inhibition of PKBa kinase in vitro. Other compounds need to undergo kinase inhibition test.
(7), this paper studies three trifluoroacetic acid and methyl chloride lutetium Cui flower two (CMDMCS) Biginelli in the reaction of methyl chlorosilane. Reaction with aldehydes / isatin compounds / acetal, ketal / 1,3- two carbonyl compounds, urea / thiourea as the starting materials, the reaction obtained in high yield 22 two hydrogen pyrimidine ketone compounds. Among them, three trifluoroacetic acid lutetium Recyclable and reused, and chloromethyl two methyl chlorosilane is cheap and thus for the preparation of the compound provides a new synthetic route, and lays a foundation for further research of the compounds.

【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R914

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