抗细胞凋亡活性化合物的构效关系研究、FAPα激活式BF211前药研究及钯催化插羰反应方法学研究

发布时间：2018-06-24 23:34

  本文选题：SP2343 + 糖尿病　； 参考：《中国科学院大学(中国科学院上海药物研究所)》2017年博士论文

【摘要】：本论文分为四部分:(1)抗胰岛β细胞凋亡活性化合物的构效关系研究;(2)抗神经细胞凋亡活性化合物的构效关系研究;(3)FAPα激活式BF211前药研究;(4)钯催化插羰反应方法学研究。糖尿病是一种严重危害人类生命健康的代谢性疾病,其中90%以上为2型糖尿病。研究发现,胰岛β细胞功能障碍是2型糖尿病发病和进展的核心,保护和恢复胰岛β细胞功能成为治疗2型糖尿病的关键,这也为新型糖尿病药物开发提供了新的方向。本课题组针对胰岛β细胞凋亡的保护,在STZ诱导的细胞模型上进行高通量筛选,得到活性化合物SP1610、SP2343和长春胺,它们不仅在细胞水平上具有抗胰岛β细胞凋亡作用,而且在STZ/HFD糖尿病小鼠上能够降血糖、降糖化血红蛋白、改善口服糖耐量、改善胰岛β细胞的受损状况。鉴于它们体内外有效,分子作用机制基本清楚,以及结构简单、分子量小、可改造空间大,可确定其为苗头化合物,进行系统的研究和开发。共设计、合成了200多个衍生物,通过体外活性评价,发现SP1610衍生物活性均未改善,研究终止;SP2343衍生物中C7体外活性明显提高,但在体内低剂量有效、高剂量有毒,而且它的成药性较差,有待进一步改善;长春胺系列仅合成了12个衍生物,发现体外活性显著提高,其中V9的活性提高了50多倍,值得进一步研究。阿尔茨海默症(AD)是当今公共健康的最大威胁之一,目前已上市药物只能缓解症状,却不能治愈。治疗AD的有效药物研发困难,主要因为其发病机制非常复杂,其中神经纤维缠结、神经细胞受损凋亡是导致病情加重的根本原因,因此,保护和恢复神经细胞功能可能是治疗AD病症的重要突破口。本课题组针对神经细胞凋亡的保护,在STZ诱导的细胞模型上进行高通量筛选,发现SP2343和SP1178不仅在细胞水平上表现出了保护活性,而且在ICV-STZ诱导的AD大鼠模型上也能够修复受损的神经细胞、减少Aβ的生成以及减轻Tau蛋白的异常磷酸化,从而改善了AD鼠的认知功能障碍,同时,对SP2343的分子作用机制也进行了深入研究。因此,可选其为值得进行深入研究和开发的苗头化合物。将已有的SP2343衍生物进行体外活性评价及构效关系研究,发现C50和C52的抗神经细胞凋亡活性提高了近80倍,基于此又继续合成50多个衍生物,但活性未进一步改善;设计合成了10个SP1178衍生物,体外活性均未明显改善,研究终止。本课题组前期选取蟾酥中抗肿瘤活性成分蟾毒灵为先导结构,通过结构优化得到了活性更强、毒性更低、成药性更好的药物候选物BF211。然而在狗上的毒性试验中发现BF211仍有较强的心脏毒性,显示无治疗窗口。至今,酶激活式靶向前体药物(enzyme-activated prodrug)在肿瘤治疗方面已取得显著的成果。其药物设计的核心思路在于运用特异性导向物修饰细胞毒性小分子,以屏蔽其活性基团,形成低活性的前药;前药进入机体内后以低活化形式被输送至靶部位,继而利用靶部位特异酶的选择性酶解作用,使得活性母体药物得以释放。成纤维细胞是肿瘤微环境中主要的间质细胞,与肿瘤细胞作用而被激活,成为肿瘤相关成纤维细胞(CAF),能高表达FAPα。FAPα属丝氨酸蛋白酶类,在90%以上的上皮癌(包括肺癌、结肠癌、乳腺癌、膀胱癌、胰腺癌、卵巢癌等)的基质活化成纤维细胞中表达。它具有促进与调控肿瘤生长,促进肿瘤的侵袭与转移的作用。基于FAPα介导的酶激活式靶向抗肿瘤前药策略已在阿霉素上成功应用,我们设计、合成了14个FAPα激活式BF211前药分子,经过酶孵育、组织孵育以及细胞毒性试验,发现前药03是低毒高效的理想分子,随后在大鼠结肠癌移植瘤模型进行体内活性评价,发现其抑瘤活性与原药接近,且毒性明显降低,表现出一定的治疗窗口。该部分工作为以后更系统的FAPα激活式前药研究及其应用奠定了基础。钯催化的插羰反应是合成羰基衍生物很常用的一类方法,如制备酰胺、酯、酸、醛、酮等。该类反应过程中最关键的步骤是引入羰基,而CO高压气体则是最常用的羰基源,但CO是剧毒气体,大量使用势必会限制此方法的广泛应用。因此,本课题组设计了两种新方法来定量释放CO以满足反应需求,从而避免大量使用CO。方法一:是利用CHCl3-CsOH?H2O体系水解定量产生CO,在钯催化下芳基碘代物与末端炔烃发生插羰偶联反应得到炔酮衍生物;方法二:是利用I2-PPh3体系配位HCOOH定量产生CO,在钯催化下进行芳基碘代物的插羰反应,再经多余HCOOH还原后得到芳醛衍生物。这两种方法具有显著优势:(1)避免了直接使用高压、高毒的CO气体;(2)利用廉价易得的原料实现了CO的定量释放以满足插羰反应需求;(3)反应条件相对温和、后处理简单、产率较高;(4)反应适用范围广泛,并在多种天然产物上成功应用。
[Abstract]:This thesis is divided into four parts: (1) study on the structure-activity relationship of anti islet beta cell apoptosis active compounds; (2) study on the structure-activity relationship of anti neuronal apoptosis active compounds; (3) FAP alpha activated BF211 prodrug study; (4) palladium catalyzed carbonylation method study. Diabetes is a metabolic disease that seriously endangers human life and health, of which 90 More than percent of type 2 diabetes. The study found that islet beta cell dysfunction is the core of the onset and progression of type 2 diabetes. Protecting and restoring islet beta cell function is the key to the treatment of type 2 diabetes, which also provides a new direction for the development of new type diabetes drugs. In the cell model, high throughput screening was carried out to obtain active compounds SP1610, SP2343 and Changchun amine. They not only inhibit the apoptosis of islet beta cells on the cell level, but also reduce blood sugar, hypoglycemic hemoglobin, improve oral glucose tolerance and improve the damage of islet beta cells in STZ/HFD diabetic mice. It is clear that the molecular mechanism is clear, and the structure is simple, the molecular weight is small and the space can be reformed. It can be determined as the compound of the seedling, and the system has been studied and developed. A total of more than 200 derivatives were designed and synthesized. Through the evaluation of the activity in vitro, the activity of SP1610 derivatives was not improved and the activity of C7 in the SP2343 derivatives was in vitro activity. It is obviously improved, but it is low dose effective in the body, high dose toxic, and its drug property is poor, it needs to be further improved; the Changchun amine series only syntheses 12 derivatives, and it is found that the activity in vitro is significantly improved, in which the activity of V9 is more than 50 times, it is worth further study. Blzheimer (AD) is the biggest threat to public health today. First, the drugs that have already been listed can only relieve symptoms but can not be cured. The difficulties in the research and development of effective drugs for the treatment of AD are mainly due to the complicated pathogenesis, in which the nerve fibers are tangled and the apoptosis of the nerve cells is the fundamental cause of the aggravation. Therefore, the protection and recovery of the function of the nerve cells may be an important process for the treatment of the disease of AD. In order to protect the apoptosis of nerve cells, our team conducted high throughput screening on the STZ induced cell model. It was found that SP2343 and SP1178 not only showed protective activity on the cell level, but also could repair damaged nerve cells in the AD rat model induced by ICV-STZ, reduce the formation of A beta and reduce the difference of Tau protein. It improves the cognitive impairment of AD mice, and further studies the molecular mechanism of SP2343. Therefore, it is optional for further research and development of the seedling compounds. The activity evaluation and structure-activity relationship of the existing SP2343 derivatives are studied in vitro, and the anti neural cell withering of C50 and C52 is found. The activity increased nearly 80 times. Based on this, the more than 50 derivatives continued to be synthesized, but the activity was not further improved; 10 SP1178 derivatives were designed and synthesized. The activity in vitro was not obviously improved, and the study was terminated. Lower sex, a better drug candidate BF211., however, in the toxicity test on dogs, BF211 still has a strong heart toxicity and shows no therapeutic window. Up to now, the enzyme activated target drug (enzyme-activated prodrug) has achieved remarkable results in the treatment of cancer. The heterosexual guide modifies the small toxic molecules of cells to shield their active groups and form low active prodrugs; the prodrugs are transported into the body and are transported to the target site in the form of low activation, and then the active mother drugs can be released by the selective enzymatic hydrolysis of target specific enzymes. Fibroblasts are the main intermediate of the tumor microenvironment. Stromal cells, activated with tumor cells, become tumor related fibroblasts (CAF), and can express high expression of FAP alpha.FAP alpha serine protease, which are expressed in the basal cells of more than 90% of the epithelial cancer (including lung, colon, breast, bladder, pancreatic, ovarian cancer, etc.). It promotes and regulates the growth of tumor. Promoting the role of tumor invasion and metastasis. The FAP alpha mediated enzyme activated target antitumor drug strategy has been successfully applied to adriamycin. We designed and synthesized 14 FAP alpha activated BF211 prodrug molecules, incubated by enzymes, tissue incubation and cytotoxicity test, and 03 were low toxic and efficient idealmolecules, followed by large amounts of prodrugs. The activity of the rat colon cancer transplanted tumor model was evaluated in vivo, and it was found that the tumor suppressor activity was close to the original drug, and the toxicity was obviously reduced, which showed a certain treatment window. This part work laid the foundation for the further research and application of the more systematic FAP alpha activating prodrug. Methods, such as the preparation of amides, esters, acids, aldehydes and ketones, the most critical step in the process is the introduction of carbonyl groups, while the CO high pressure gas is the most commonly used carbonyl source, but CO is a highly toxic gas. A large number of use is bound to limit the widespread use of this method. Therefore, two new methods have been set up to quantify the release of CO to meet response needs. In order to avoid a large number of use of CO. method one: using the CHCl3-CsOH? H2O system to hydrolyze the quantitative production of CO, and the coupling reaction of the aryl iodides with the terminal alkynes under the catalysis of palladium to get the alkynone derivative; method two: using the I2-PPh3 system coordination HCOOH to produce CO, the aryl iodide intercalation reaction under the catalysis of palladium, and then many more After the reduction of residual HCOOH, the aromatic aldehyde derivatives were obtained. These two methods have significant advantages: (1) avoiding the direct use of high pressure, high toxic CO gas; (2) using cheap and easy available raw materials to achieve the quantitative release of CO to meet the carbonyl reaction demand; (3) the reaction conditions are relatively mild, the post-processing is simple, the yield is high; (4) the application range is wide, and A variety of natural products have been successfully applied.
【学位授予单位】：中国科学院大学(中国科学院上海药物研究所)
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R914
，

本文编号：2063487