本文選題：手性 + 化學合成��； 參考：《山東大學》2017年博士論文

【摘要】：1.手性亞砜類GPR40激動劑的設計、合成及活性研究研究背景糖尿病是影響人類健康的重大代謝性疾病,其中二型糖尿病占90%以上。GPR40近年來被發(fā)現(xiàn)是治療二型糖尿病的新型靶點。根據(jù)該靶點的作用機制,人們發(fā)現(xiàn)其促胰島素分泌作用是葡萄糖依賴性的。因此GPR40激動劑作為降糖藥物可以避免傳統(tǒng)降糖藥物(如磺胺類)容易引起低血糖的副作用,降低低血糖風險。GPR40已經成為近年來降糖藥物研究中的熱門靶標。目標化合物的設計、合成、活性篩選近年來,大量GPR40激動劑被發(fā)現(xiàn)并報道,多個具有開發(fā)價值的小分子處于藥物研發(fā)的不同階段。TAK-875雖然因為肝毒性而停止了臨床試驗,但是它仍然是到現(xiàn)在為止唯一一個進入到三期臨床試驗的、口服有效的GPR40激動劑。本課題對TAK-875進行結構改造,根據(jù)生物電子等排原理,將TAK-875的砜基團轉換成亞砜基團,以較小的結構變化來保持化合物原有的生物活性。此外,同系物中亞砜類化合物的極性通常要大于砜類化合物。因此,TAK-875的亞砜類似物的水溶性會得到提高,在理論上降低脂毒性,同時藥物代謝動力學性質會得到改善,進而提高化合物的體內活性。結構改造得到的化合物中亞砜基團為手性基團,本課題進一步考察了其不同立體構型對化合物活性的影響。在合成工作中,本課題首先合成了 TAK-875作為對照藥物;其次,利用合成對照藥物的適當中間體經親核取代反應、還原反應、Mitsunobu反應、氧化反應、水解反應得到了 TAK-875的亞砜消旋體衍生物(g24);第三,手性制備拆分消旋體亞砜衍生物g24,得到光學純的差向異構體亞砜產物g25(,S 100.0%de)和g26(R,S,100.0%de)。最后,采用不對稱氧化的方法制備了較大量的較高光學純度的亞砜衍生物。檢測了目標化合物的極性和水溶性。通過Ca2+流量分析實驗,測定了目標化合物的EC50。建立大鼠糖尿病模型,進行口服糖耐量試驗(OGTT)。通過藥物代謝動力學試驗和體內外肝毒性試驗,考察目標化合物的成藥性。結果本研究合成的中間體和目標化合物由IR、1H NMR、1C NMR、ESI-MS、HR-MS進行了基本結構確定。測試了化合物的旋光度。化合物的光學純度由手性分析HPLC確定。根據(jù)文獻報道和圓二色譜的方法確定了不同目標化合物的立體構型。經Scifinder等文獻檢索工具檢索證實,所有目標化合物均為新型化合物,未見文獻報道。目標化合物體外保持了對照藥物的活性,g29(主要含有g25)降糖效果優(yōu)于對照藥物。藥物代謝動力學實驗發(fā)現(xiàn)g25(S,S)更容易被吸收,在體內可以長時間保持有效濃度。相比g26(R,S,g25(S,S)更容易代謝成對照藥物。在糖尿病大鼠模型中口服給藥一個月后,經血生化檢測發(fā)現(xiàn),g25(S,S)用藥組大鼠的谷丙轉氨酶和谷草轉氨酶指標比對照藥物組要高。通過對HL7702細胞凋亡試驗,發(fā)現(xiàn)g25(S,S)可引起肝細胞凋亡,且比對照藥物的作用要強。結論將TAK-875的砜基轉換成亞砜基保留了該類化合物的體外活性,這可由TAK-875與hGPR40的結晶復合物的結合模式解釋。而且,亞砜類似物水溶性顯著增加。g25(S)在大鼠體內的藥效以及藥動優(yōu)于TAK-875,且容易代謝成TAK-875,因此可能具有潛在的肝毒性。該假設在糖尿病大鼠血的生化檢測中得到了證實。g25(S,S)可以引起HL7702細胞凋亡,進一步表明其肝毒性。因此,肝毒性是未來GPR40激動劑設計時應當充分考慮的問題。其次,將砜基改造成亞砜基是藥物設計中的一把"雙刃劍",其生物電子等排體替換對母體藥物的成藥性具有很大的依賴性。如果母體化合物成藥性良好或為上市藥物,隨之的亞砜衍生物研發(fā)成功率較大。如果母體藥物研發(fā)失敗,亞砜衍生物研發(fā)就很有可能隨之失敗。2.手性亞砜類MMP-9抑制劑的設計、合成及活性研究研究背景糖尿病可以引發(fā)多種并發(fā)癥,糖尿病傷口難以愈合就是其中之一,迄今也沒有找到有效治療方法。近年來發(fā)現(xiàn)MMP-9在糖尿病傷口中表達升高,抑制MMP-9有利糖尿病傷口的愈合。因此,研發(fā)MMP-9的選擇性抑制劑將給這類疾病的治療帶來機會。目標化合物的設計、合成、活性篩選根據(jù)文獻報道,ND-322和ND-364是抑制MMP-9活性較好的兩個小分子化合物。但是,這兩個化合物都是消旋體。于是分別合成ND-322和ND-364的(R)和(S)對映異構體,以考查環(huán)硫乙烷部分不同立體構型對MMP-9抑制活性的影響。ND-364為ND-322的體內N-乙�；x產物,對MMP-9的活性更好,本課題將其砜基改造為亞砜基,考查手性亞砜結構對活性的影響。另外,將苯環(huán)上的伯氨基BOC保護,考察該處取代基大小對活性的影響。以光學純的縮水甘油為手性源起始原料,經氯磺化反應得到的中間體,與苯硫酚反應后,立體結構保持。在Cs2CO2存在的情況下進行成醚反應,同時完成了關環(huán)反應;硝基在酸性條件下鋅粉還原后,過濾鋅粉,溶液調PH至堿性直接BOC保護或乙�；�;之后經氧化,硫代,或脫保護得到目標產物。硫代反應立體構型反轉。另外,適當中間體經不對稱氧化,得到了不同立體構型的亞砜產物,與硫脲反應得到目標產物。目標化合物體外測試了對MMP-2、MMP-9、APN和HDACs的抑制活性。結果所合成的目標化合物由IR、1HNMR、ESI-MS、HR-MS進行了基本結構確定。測試了化合物的旋光度。化合物的光學純度由手性分析HPLC確定。根據(jù)文獻報道和圓二色譜的方法確定了不同目標化合物的立體構型。經Scifinder等文獻檢索工具檢索證實,所有目標化合物均為新型化合物,未見文獻報道。化合物na8、nb8、na11和nb11表現(xiàn)出對MMP-2和MMP-9良好的選擇性抑制,與參考藥物ND-322和ND-364相比,活性得到保留。結論環(huán)硫乙烷部分的立體構型對活性幾乎沒有影響。亞砜改造顯著降低了化合物的活性,根據(jù)此類化合物獨特的結合機理,該部分有強吸電子基團有利于活性提高。與苯胺連接的基團不宜過大,否則因空間位阻的原因而使化合物喪失抑酶活性。這些規(guī)律的發(fā)現(xiàn)對下一步發(fā)現(xiàn)高效選擇性的MMP-9抑制劑具有參考價值。
[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.
【學位授予單位】：山東大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：R914;R96
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本文編號：1988661