SGLT2抑制剂的设计、合成与生物活性研究

发布时间：2018-04-14 14:22

本文选题：SGLT2抑制剂 + Dapagliflozin　；参考：《天津医科大学》2014年博士论文

【摘要】：目的:Ⅱ型糖尿病是一种以胰岛素相对不足和血糖过高为主要特征的慢性代谢疾病,能够诱发一系列并发症。由于发病机制复杂,病程长,目前的治疗药物难以有效控制血糖水平,因此临床上迫切需要具有全新作用机制的新型降糖药。原尿中99%的葡糖糖流经肾小管时被重吸收,其重吸收主要依赖于两种钠-葡萄糖协同转运蛋(sodium-dependent glucose cotrans-porters,SGLTs)介导:SGLT2 起主要作用,转运重吸收葡萄糖的90%;SGLT1转运重吸收葡萄糖的10%。选择性地抑制SGLT2的活性,能够减少葡萄糖的重吸收,增加尿糖,降低血糖水平,已成为治疗Ⅱ型糖尿病的的新型靶标。目前报道的SGLT2抑制剂主要是O-芳基糖苷抑制剂、C-芳基糖苷抑制剂,O-芳基糖苷抑制剂选择性较低、代谢不稳定,而C-芳基糖苷抑制剂克服了以上不足之处,取得了巨大的成绩,其中Dapagliflozin和Canagflozin两种SGLT2抑制剂已分别在欧洲和美国上市。我们以Dapagliflozin为先导化合物,设计、合成新型SGLT2抑制剂并进行活性筛选,期望能够获得新的有效的的SGLT2抑制剂。方法:我们深刻分析Dapagliflozin、Empagliflozin和Canagliflozin等C-芳基抑制剂的分子结构,发现这些化合物的结构具有很大的相似性,即分子左端为一个葡萄糖片断,该葡萄糖片断通过C-糖苷键与一个苯环相连,该苯环的间位通过—个亚甲基与另外一个芳基相连。针对上述SGLT2抑制剂尤其是Dapagliflozin的分子结构特点,我们设计了三类化合物:单甲基C-碳糖苷类、偕二甲基C-碳糖苷类和环丙基C-碳糖苷类,三类化合物均以Dapagliflozin作为阳性对照药,进行了体内抗血糖活性评价,部分化合物还进行了体外活性测试。对实验室合成的SGLT2抑制剂D6进行逆合成分析:发现化合物D6有两个结构特点:一是糖环部分的6位-去氧,二是碳糖苷,所以6-脱氧和碳糖苷是合成D6的关键步骤。考虑到生产需要和专利避开,我们以先构建碳糖苷,后构建6-去氧为策略,采用汇聚式的合成方法,经过多次试验最终确定了 D6的新的合成路线。结果:通过汇聚式的合成路线合成目标化合物。在第1-3章节中,以取代的苯甲酸和葡萄糖酸内酯为起始物,经过酰氯化或酯化、傅克反应、Simmons-Smith反应、溴-锂交换、三乙基硅烷还原、乙酰化和脱乙酰化等步骤合成了单甲基类化合物3个、偕二甲基类化合物3个、环丙基类化合物7个,化合物通过IH-NMR、13C-NMR和HR-MS、IR表征了它们的结构,体内活性测试发现它们均有较好的抗高血糖活性,部分药物具有一定的成药性。我们以碘化物为起始原料,经过催化氢化、酸性催化水解、Swern氧化、溴-锂交换、苄基脱保护反应成功地合成D6,并对催化氢化、酸性催化水解和苄基脱保护等步骤进行反复试验和多次优化,找到了一条产率高、易操作、重复性强的合成D6的工艺路线,并且发现SrCl_2对甲基糖苷的酸性水解具有极为有效的催化作用,为D6的药理、毒理等实验提供了有力的原料保障。结论:论文结合设计合成的三类C-芳基糖苷类SGLT2抑制剂的化学结构和药理活性,发现它们的活性强弱为:单甲基类活性强于环丙基类,再强于偕二甲基类,即在SGLT2抑制剂的两芳基之间亚甲基处添加基团活性保留,且基团越大活性越弱,苯环上的取代基以氯最优。这是学术上首次系统和科学地阐明了SGLT2抑制剂结构中的两苯环之间亚甲基上的构效关系,是对SGLT2抑制剂构效关系研究的重要补充,在此研究基础上我们有可能发现更好的成药分子。开发的D6的新路线开辟了合成各类脱氧C-糖苷的一种通用方法;该新合成工艺,具有稳定性好和易操作的优点,并具有自主知识产权,目前已被实验室中试采用,已在公斤级中试平台上运行四批,稳定可靠,为D6的临床前研究提供了有力的物质保障。在学术界首次发现甲基糖苷水解共催化剂SrCl_2,高效、绿色、可控,现已在实验室的小规模实验中广泛使用,并在公斤级平台上得到了验证。
[Abstract]:Objective: diabetes is a relative lack of insulin and hyperglycemia is the main characteristic of chronic metabolic diseases, can cause a series of complications. Because the pathogenesis is complex, long course of disease, the treatment is difficult to effectively control the blood glucose level, therefore clinically urgent need for new antidiabetic drugs with new mechanisms of action. 99% of the glucose in urine flow tubular reabsorption is, its reabsorption mainly depends on two kinds of sodium glucose co transporter (sodium-dependent glucose, cotrans-porters, SGLTs) - mediated: SGLT2 plays a major role, transport of glucose reabsorption 90%; SGLT1 transport of glucose reabsorption of 10%. selectively inhibits the activity of SGLT2, to reduce glucose reabsorption and increase urine glucose, reduce blood sugar levels, has become a new target for the treatment of type II diabetes. The reported SGLT2 inhibitor is O- aryl Glucosidase inhibitors, C- aryl glucoside inhibitors, O- aryl glucoside inhibitors lower selectivity, metabolic instability, and C- aryl glucoside inhibitors to overcome the above shortcomings, has made great achievements, including Dapagliflozin and Canagflozin two kinds of SGLT2 inhibitors have been respectively in European and American market. We design with Dapagliflozin as the leading compound,, synthesis of novel SGLT2 inhibitors and activity screening, is expected to be a new effective inhibitor of SGLT2. Methods: We deeply analyze Dapagliflozin, Empagliflozin and Canagliflozin, the molecular structure of C- aryl inhibitors, found that the structures of these compounds are very similar, which is a molecular fragment of glucose, the glucose fragment connected with a benzene ring by C- glycosidic bond between the benzene ring, through a methylene linked with another aryl. Based on the above SGLT2 Inhibitor especially the molecular structure characteristics of Dapagliflozin, we designed three compounds: single methyl glucoside C- carbon, with two methyl carbon C- glycosides and cyclopropyl C- carbon glycosides, three compounds with Dapagliflozin as positive control medicine, the in vivo activity of glucose evaluation, some compounds were tested in vitro activity test. The SGLT2 inhibitor D6 synthesis laboratory for inverse synthetic analysis: we found that compound D6 has two structural characteristics: one is the sugar moiety 6 - deoxy, two carbon glycoside, so 6- deoxidation and carbon glycoside is a key step in the synthesis of D6. Considering the production needs and avoid the first US patent. After the construction of the carbon glycoside, construct 6- DNA as the strategy, the synthesis method is convergent, after many tests finally identified a new synthetic route of D6. Results: the compounds synthesized by convergent goal in section 1-3. In the chapter, using benzoic acid and gluconic acid lactone substituted as the starting material, after esterification or acylation, Friedel Crafts reaction, Simmons-Smith reaction, bromine lithium exchange, three ethyl silane reduction, acetylation and deacetylation and other steps of the synthesis of methyl compounds 3, 3 with two methyl compounds. Cyclopropyl compounds 7 were characterized by IH-NMR, 13C-NMR, IR and HR-MS, their structures were characterized, in vivo activity test found that they have good anti hyperglycemic activity, drug resistance has become a part of us. With iodide as the starting material, through catalytic hydrogenation, acid catalyzed hydrolysis, oxidation of Swern, lithium bromide exchange, benzyl deprotected was successfully synthesized D6 and catalytic hydrogenation, the acid catalyzed hydrolysis and benzyl based protection steps were repeated testing and optimization, and found a high yield, easy operation, strong repeatability of the synthesis of D6. Route, and found that the SrCl_2 of methyl glucoside acid hydrolysis is extremely effective in catalysis, D6 pharmacology, provides a strong material guarantee for toxicological experiments. Conclusion: the combination of chemical structure and drug design and synthesis of three kinds of C- aryl glucoside SGLT2 inhibitor activity, that activity of their for single class activity stronger than methyl cyclopropane base, strong in two with methyl, which is between two aryl methylene groups add SGLT2 inhibitor activity retention, and the activity of the weak group, the substituent on the benzene ring with chlorine. This is the first optimal academic system and scientifically expound the structure-activity relationship between the methylene SGLT2 inhibitor in the structure of two benzene rings, is an important complement to the structure-activity relationship of SGLT2 inhibitors, on the basis of this study, we may find a better route. New molecular medicine development D6 development A general method for synthesis of deoxy C- glucoside; the new synthetic technology, has the advantages of good stability and easy operation, and has independent intellectual property rights, has been used in the laboratory test, test platform has operated four batches in kilograms in stable and reliable for clinical D6 study provides strong material. In the academic circles for the first time found methyl glycoside hydrolysis catalyst is SrCl_2, green, efficient, controllable, is now widely used in small scale experiments in the laboratory, and has been verified in the kilo platform.

【学位授予单位】：天津医科大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R914;R96

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