基于结构的Plk1选择性抑制剂的设计、合成及抗肿瘤活性研究

发布时间：2019-07-04 21:23

【摘要】：Polo样激酶(polo-like kinase, Plk)是一类广泛存在于真核生物中的丝氨酸/苏氨酸蛋白激酶。其中,Plkl调控细胞有丝分裂多个环节,在80%人类肿瘤细胞中过度表达,包括乳腺癌、结肠直肠癌、前列腺癌、胰腺癌、非小细胞肺癌、卵巢癌等,但在正常组织中很少表达。抑制Plkl的活性可以产生良好的抗癌效果,因而Plkl是一个很好的抗肿瘤药物研发的靶点。目前,已有众多Plkl小分子靶向抑制剂被报道,但仍存在着Plk家族内部选择性差、药代动力学方面的诸多问题,开发特异性、可口服的Plkl抑制剂因此至关重要。本研究采用基于结构的药物设计理论,以首个进入临床实验的Plkl抑制剂BI2536为先导化合物,保留其核心药效团骨架,利用家族内部氨基酸残基的细微差异,进行特异性的结构修饰从而达到提高活性与选择性的目的,再利用计算机辅助软件进行虚拟评价,设计并合成了27个结构新颖的全新化合物。最后,采用体外激酶抑制实验与抗肿瘤细胞株实验对化合物进行生物学评价。测试结果表明,所设计的大部分化合物均对Plkl产生了低纳摩尔浓度的抑制活性以及家族内部的选择性。其中化合物C2对Plkl的ICso值达到了4.134 nM,优于阳性对照物BI2536和十字孢碱,并且将家族内部Plk2、Plk3的选择性分别提高至54倍和35倍。同时该化合物对体外MCF-7、K562、Hela癌细胞株的抑制活性分别为40.69 nM、41.67 nM、37.71 nM,同样优于阳性对照物BI 2536与紫杉醇。本课题初步筛选出了一系列高效、低毒、结构新颖的Plkl抑制剂,为进一步开发新型高效的抗癌药物奠定了基础。
文内图片：

图１．２逦（ａ）抑制剂与ＮＣＤ邋口袋的结合模式；（ｂ）抑制剂与ＰＢＤ邋口袋的结合模式逡逑Ｆｉｕｒｅ邋１．２ａＴｈｅ邋ｂｉｎｄｉｎｍｏｄｅ邋ｏｆ邋ｉｎｈ化ｉｏｒ邋ｗｉｈ邋化ｅ邋ＮＣＤｂＴｈｅ邋ｂｉｎｄｉｎｍｏｄｅ邋ｏｆ邋ｉｎｈ化ｉｔｏｒ邋ｗｉｔｈ逡逑

图片说明：图１．２逦（ａ）抑制剂与ＮＣＤ邋口袋的结合模式；（ｂ）抑制剂与ＰＢＤ邋口袋的结合模式逡逑Ｆｉｕｒｅ邋１．２ａＴｈｅ邋ｂｉｎｄｉｎｍｏｄｅ邋ｏｆ邋ｉｎｈ化ｉｏｒ邋ｗｉｈ邋化ｅ邋ＮＣＤｂＴｈｅ邋ｂｉｎｄｉｎｍｏｄｅ邋ｏｆ邋ｉｎｈ化ｉｔｏｒ邋ｗｉｔｈ逡逑
[Abstract]:Polo-like kinase (polo-like kinase, Plk) is a kind of serine / threonine protein kinase which widely exists in eukaryotes. Among them, Plkl regulates mitosis and is overexpressed in 80% of human tumor cells, including breast cancer, colorectal cancer, prostate cancer, pancreatic cancer, non-small cell lung cancer, ovarian cancer and so on, but rarely expressed in normal tissues. Inhibition of Plkl activity can produce good anticancer effect, so Plkl is a good target for antitumor drug development. At present, many Plkl small molecule targeting inhibitors have been reported, but there are still many problems in Plk family, such as poor selectivity and pharmacokinetics. It is very important to develop specific and oral Plkl inhibitors. In this study, the structure-based drug design theory was used, the first Plkl inhibitor BI2536, which entered the clinical trial, was used as the lead compound to retain its core pharmacophore skeleton, and the specific structure modification was carried out by using the nuances of amino acid residues within the family to improve the activity and selectivity. 27 new compounds with novel structure were designed and synthesized by virtual evaluation with computer-aided software. Finally, the biological evaluation of the compounds was carried out by in vitro kinase inhibition test and anti-tumor cell line test. The results showed that most of the compounds produced low nanomolconcentration inhibitory activity and intra-family selectivity to Plkl. The ICSO value of compound C2 to Plkl was 4.134 nM, which was better than that of BI2536 and Cruciferine, and the selectivity of Plk2,Plk3 in the family was 54 times and 35 times, respectively. At the same time, the inhibitory activity of the compound on MCF-7,K562,Hela cell line in vitro was 40.69 nM,41.67 nM,37.71 nM, which was also better than that of BI 2536 and paclitaxel, respectively. In this paper, a series of Plkl inhibitors with high efficiency, low toxicity and novel structure were screened out, which laid a foundation for the further development of new and efficient anticancer drugs.
【学位授予单位】：合肥工业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R914;R96

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