手性亚砜类GPR40激动剂与MMP-9抑制剂的设计、合成及活性研究

发布时间：2018-06-06 23:50

  本文选题：手性 + 化学合成　； 参考：《山东大学》2017年博士论文

【摘要】：1.手性亚砜类GPR40激动剂的设计、合成及活性研究研究背景糖尿病是影响人类健康的重大代谢性疾病,其中二型糖尿病占90%以上。GPR40近年来被发现是治疗二型糖尿病的新型靶点。根据该靶点的作用机制,人们发现其促胰岛素分泌作用是葡萄糖依赖性的。因此GPR40激动剂作为降糖药物可以避免传统降糖药物(如磺胺类)容易引起低血糖的副作用,降低低血糖风险。GPR40已经成为近年来降糖药物研究中的热门靶标。目标化合物的设计、合成、活性筛选近年来,大量GPR40激动剂被发现并报道,多个具有开发价值的小分子处于药物研发的不同阶段。TAK-875虽然因为肝毒性而停止了临床试验,但是它仍然是到现在为止唯一一个进入到三期临床试验的、口服有效的GPR40激动剂。本课题对TAK-875进行结构改造,根据生物电子等排原理,将TAK-875的砜基团转换成亚砜基团,以较小的结构变化来保持化合物原有的生物活性。此外,同系物中亚砜类化合物的极性通常要大于砜类化合物。因此,TAK-875的亚砜类似物的水溶性会得到提高,在理论上降低脂毒性,同时药物代谢动力学性质会得到改善,进而提高化合物的体内活性。结构改造得到的化合物中亚砜基团为手性基团,本课题进一步考察了其不同立体构型对化合物活性的影响。在合成工作中,本课题首先合成了 TAK-875作为对照药物;其次,利用合成对照药物的适当中间体经亲核取代反应、还原反应、Mitsunobu反应、氧化反应、水解反应得到了 TAK-875的亚砜消旋体衍生物(g24);第三,手性制备拆分消旋体亚砜衍生物g24,得到光学纯的差向异构体亚砜产物g25(,S 100.0%de)和g26(R,S,100.0%de)。最后,采用不对称氧化的方法制备了较大量的较高光学纯度的亚砜衍生物。检测了目标化合物的极性和水溶性。通过Ca2+流量分析实验,测定了目标化合物的EC50。建立大鼠糖尿病模型,进行口服糖耐量试验(OGTT)。通过药物代谢动力学试验和体内外肝毒性试验,考察目标化合物的成药性。结果本研究合成的中间体和目标化合物由IR、1H NMR、1C NMR、ESI-MS、HR-MS进行了基本结构确定。测试了化合物的旋光度。化合物的光学纯度由手性分析HPLC确定。根据文献报道和圆二色谱的方法确定了不同目标化合物的立体构型。经Scifinder等文献检索工具检索证实,所有目标化合物均为新型化合物,未见文献报道。目标化合物体外保持了对照药物的活性,g29(主要含有g25)降糖效果优于对照药物。药物代谢动力学实验发现g25(S,S)更容易被吸收,在体内可以长时间保持有效浓度。相比g26(R,S,g25(S,S)更容易代谢成对照药物。在糖尿病大鼠模型中口服给药一个月后,经血生化检测发现,g25(S,S)用药组大鼠的谷丙转氨酶和谷草转氨酶指标比对照药物组要高。通过对HL7702细胞凋亡试验,发现g25(S,S)可引起肝细胞凋亡,且比对照药物的作用要强。结论将TAK-875的砜基转换成亚砜基保留了该类化合物的体外活性,这可由TAK-875与hGPR40的结晶复合物的结合模式解释。而且,亚砜类似物水溶性显著增加。g25(S)在大鼠体内的药效以及药动优于TAK-875,且容易代谢成TAK-875,因此可能具有潜在的肝毒性。该假设在糖尿病大鼠血的生化检测中得到了证实。g25(S,S)可以引起HL7702细胞凋亡,进一步表明其肝毒性。因此,肝毒性是未来GPR40激动剂设计时应当充分考虑的问题。其次,将砜基改造成亚砜基是药物设计中的一把"双刃剑",其生物电子等排体替换对母体药物的成药性具有很大的依赖性。如果母体化合物成药性良好或为上市药物,随之的亚砜衍生物研发成功率较大。如果母体药物研发失败,亚砜衍生物研发就很有可能随之失败。2.手性亚砜类MMP-9抑制剂的设计、合成及活性研究研究背景糖尿病可以引发多种并发症,糖尿病伤口难以愈合就是其中之一,迄今也没有找到有效治疗方法。近年来发现MMP-9在糖尿病伤口中表达升高,抑制MMP-9有利糖尿病伤口的愈合。因此,研发MMP-9的选择性抑制剂将给这类疾病的治疗带来机会。目标化合物的设计、合成、活性筛选根据文献报道,ND-322和ND-364是抑制MMP-9活性较好的两个小分子化合物。但是,这两个化合物都是消旋体。于是分别合成ND-322和ND-364的(R)和(S)对映异构体,以考查环硫乙烷部分不同立体构型对MMP-9抑制活性的影响。ND-364为ND-322的体内N-乙酰化代谢产物,对MMP-9的活性更好,本课题将其砜基改造为亚砜基,考查手性亚砜结构对活性的影响。另外,将苯环上的伯氨基BOC保护,考察该处取代基大小对活性的影响。以光学纯的缩水甘油为手性源起始原料,经氯磺化反应得到的中间体,与苯硫酚反应后,立体结构保持。在Cs2CO2存在的情况下进行成醚反应,同时完成了关环反应;硝基在酸性条件下锌粉还原后,过滤锌粉,溶液调PH至碱性直接BOC保护或乙酰化;之后经氧化,硫代,或脱保护得到目标产物。硫代反应立体构型反转。另外,适当中间体经不对称氧化,得到了不同立体构型的亚砜产物,与硫脲反应得到目标产物。目标化合物体外测试了对MMP-2、MMP-9、APN和HDACs的抑制活性。结果所合成的目标化合物由IR、1HNMR、ESI-MS、HR-MS进行了基本结构确定。测试了化合物的旋光度。化合物的光学纯度由手性分析HPLC确定。根据文献报道和圆二色谱的方法确定了不同目标化合物的立体构型。经Scifinder等文献检索工具检索证实,所有目标化合物均为新型化合物,未见文献报道。化合物na8、nb8、na11和nb11表现出对MMP-2和MMP-9良好的选择性抑制,与参考药物ND-322和ND-364相比,活性得到保留。结论环硫乙烷部分的立体构型对活性几乎没有影响。亚砜改造显著降低了化合物的活性,根据此类化合物独特的结合机理,该部分有强吸电子基团有利于活性提高。与苯胺连接的基团不宜过大,否则因空间位阻的原因而使化合物丧失抑酶活性。这些规律的发现对下一步发现高效选择性的MMP-9抑制剂具有参考价值。
[Abstract]:Design, synthesis and activity study of 1. chiral sulfoxide GPR40 agonists, diabetes is a major metabolic disease affecting human health. Among them, type two diabetes, which accounts for more than 90% of.GPR40, has been found to be a new target for the treatment of type two diabetes in recent years. Glucose dependence. Therefore, as a hypoglycemic agent, GPR40 agonists can avoid the side effects of traditional hypoglycemic drugs (such as sulfonamides) that cause hypoglycemia and reduce the risk of hypoglycemia,.GPR40 has become a hot target in the study of hypoglycemic drugs in recent years. The design, synthesis, and activity screening of target compounds in recent years, a large number of GPR40 agonists It was found and reported that several small molecules of value in development were in the different stages of drug development.TAK-875, although the clinical trials were stopped because of liver toxicity, but it is still the only one that has now entered the three phase of clinical trials, oral effective GPR40 agonists. The sulfone group of TAK-875 is converted into a sulfoxide group to maintain the original biological activity of the compound with smaller structural changes. In addition, the polarity of the sulfoxide compounds in the homologues is usually greater than the sulfone. Therefore, the water solubility of the TAK-875's sulfoxide analogues will be improved, and the toxicity of low fat is reduced in theory. The kinetic properties of drug metabolism will be improved and the activity of compounds in vivo is improved. The sulfoxide group of the compound is a chiral group. In this subject, the effect of different stereotypes on the activity of compounds is further investigated. In the synthesis work, TAK-875 was first synthesized as a control drug; secondly, profit. TAK-875's sulfoxide racemate derivative (G24) was obtained by nucleophilic substitution reaction, reduction reaction, Mitsunobu reaction, oxidation reaction and hydrolysis reaction with appropriate intermediates of synthetic control drugs. Third, chiral preparation of racemo sulfoxide derivative G24 was obtained, and the optical pure differential isomer sulfoxide products G25 (S 100.0%de) and g26 (R, S, 100.0%de) were obtained. Finally, a large number of high optical purity sulfoxide derivatives were prepared by asymmetric oxidation. The polarity and water solubility of the target compounds were detected. The diabetic rat model of the target compound was established by the Ca2+ flow analysis experiment, and the oral glucose tolerance test (OGTT) was carried out. The pharmacokinetic test of the drug was carried out. The properties of the target compounds were investigated by the test of hepatotoxicity in vivo and in vitro. Results the intermediate and target compounds synthesized by this study were determined by IR, 1H NMR, 1C NMR, ESI-MS, HR-MS. The optical purity of the compound was tested. The optical purity of the compound was determined by the chiral analytical HPLC. According to the literature and the round two chromatography method, The stereotyping of different target compounds was determined. It was confirmed by Scifinder and other literature retrieval tools that all the target compounds were new compounds, and no literature was reported. The target compound kept the activity of the control drug in vitro, and the effect of g29 (mainly containing G25) was better than that of the drug. The drug metabolic kinetics experiment found that G25 (S, S) was more effective. It is easy to be absorbed and maintain effective concentration in the body for a long time. Compared with g26 (R, S, G25 (S, S), it is easier to replace the control drug. After a month of oral administration in the diabetic rat model, the blood biochemical test found that the glutamic aminotransferase and glutamic pyruvic transaminase index in G25 (S, S) rats are higher than those of the control group. Through HL7702, the HL7702 is fine. It was found that G25 (S, S) could cause hepatocyte apoptosis and was stronger than the control drug. Conclusion the sulfone base of TAK-875 was converted into sulfoxyl group and retained the in vitro activity of the compounds, which could be explained by the binding mode of the crystalline complex of TAK-875 and hGPR40. Moreover, the water solubility of the sulfoxide analogue significantly increased the.G25 (S) in the rat body. It is suggested that.G25 (S, S) can cause apoptosis of HL7702 cells in the biochemical test of blood of diabetic rats and further demonstrate its hepatotoxicity. Therefore, liver toxicity should be fully considered when designing GPR40 agonists in the future. Therefore, liver toxicity should be fully considered in the design of GPR40 agonists in the future. Secondly, the sulfonyl group is transformed into a sulfoxyl group, which is a "double-edged sword" in the drug design. The replacement of biological electrons is very dependent on the drug properties of the parent drug. If the parent compound is well prepared or listed as a drug, the subsequent development of the sulfoxide derivative is more successful. If the mother drug is not developed, The development of sulfoxide derivatives is likely to fail in the design, synthesis and activity study of the.2. sulfoxide MMP-9 inhibitors. Background diabetes can lead to a variety of complications. The difficulty of healing of diabetic wounds is one of them, and so far no effective treatment has been found. In recent years, the expression of MMP-9 in diabetic wounds has been found to be elevated. Inhibition of the healing of MMP-9 beneficial diabetic wounds. Therefore, the development of selective inhibitors of MMP-9 will bring opportunities for the treatment of these diseases. The design, synthesis, and activity screening of target compounds are reported in the literature, ND-322 and ND-364 are two small molecules that inhibit MMP-9 activity better. However, these two compounds are racemes. The effects of the synthesis of ND-322 and ND-364 (R) and (S) enantiomers respectively, to examine the effects of different stereotypes of cyclo ethane on the inhibitory activity of MMP-9,.ND-364 is the metabolite of N- acetylation in the body of ND-322, and the activity is better for MMP-9. The sulfone group is transformed into sulfoxyl group and the effect of the structure of the chiral sulfoxide on the activity is investigated. In addition, the effect of the structure of the sulfoxide on the activity of the sulfone is investigated. The effect of the primary amino BOC on the benzene ring is protected. The effect of the size of the substituent on the activity is investigated. The intermediate of the optical pure glycidyl is used as the starting material of the chiral source, the intermediate of the chlorulphonation reaction and the reaction with benzenes are maintained. The reaction of the ether is carried out in the presence of Cs2CO2, and the ring reaction is completed at the same time; the nitro is in the acid strip. After the reduction of zinc powder, the zinc powder is filtered, the solution is adjusted to the alkaline direct BOC protection or acetylation, and then the target products are obtained by oxidation, thiosulfate, or deprotection. The stereotyping of the thiosulfate reaction is reversed. In addition, the appropriate intermediate is oxidized by asymmetric oxidation to obtain the product of the sulfoxide with different stereotypes, and the target product is obtained with thiourea. Target synthesis is obtained. The inhibitory activity of MMP-2, MMP-9, APN and HDACs was tested outside the object. Results the synthesized target compounds were determined by IR, 1HNMR, ESI-MS, HR-MS. The optical purity of the compound was tested. The optical purity of the compound was determined by the chiral analysis of HPLC. The different target compounds were determined by the methods of literature and circular two chromatography. The stereotyping was confirmed by Scifinder and other literature retrieval tools. All the target compounds were new compounds, and no literature was reported. Compounds na8, NB8, na11 and nb11 showed good selective inhibition to MMP-2 and MMP-9 and retained the viability compared with the reference drugs ND-322 and ND-364. Conclusion the stereotyping of the part of cyclo ethane was concluded. It has little effect on activity. Sulfone transformation significantly reduces the activity of compounds. According to the unique binding mechanism of such compounds, the part has a strong electron absorbing group which is beneficial to the activity. The group that is connected with aniline should not be too large, otherwise the compounds will lose the enzyme activity due to the steric hindrance. These rules are found right down. One step is to find highly selective MMP-9 inhibitors with reference value.
【学位授予单位】：山东大学
【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：1988661