本文選題：SP2343 + 糖尿病��； 參考：《中國科學(xué)院大學(xué)(中國科學(xué)院上海藥物研究所)》2017年博士論文

【摘要】：本論文分為四部分:(1)抗胰島β細(xì)胞凋亡活性化合物的構(gòu)效關(guān)系研究;(2)抗神經(jīng)細(xì)胞凋亡活性化合物的構(gòu)效關(guān)系研究;(3)FAPα激活式BF211前藥研究;(4)鈀催化插羰反應(yīng)方法學(xué)研究。糖尿病是一種嚴(yán)重危害人類生命健康的代謝性疾病,其中90%以上為2型糖尿病。研究發(fā)現(xiàn),胰島β細(xì)胞功能障礙是2型糖尿病發(fā)病和進(jìn)展的核心,保護(hù)和恢復(fù)胰島β細(xì)胞功能成為治療2型糖尿病的關(guān)鍵,這也為新型糖尿病藥物開發(fā)提供了新的方向。本課題組針對胰島β細(xì)胞凋亡的保護(hù),在STZ誘導(dǎo)的細(xì)胞模型上進(jìn)行高通量篩選,得到活性化合物SP1610、SP2343和長春胺,它們不僅在細(xì)胞水平上具有抗胰島β細(xì)胞凋亡作用,而且在STZ/HFD糖尿病小鼠上能夠降血糖、降糖化血紅蛋白、改善口服糖耐量、改善胰島β細(xì)胞的受損狀況。鑒于它們體內(nèi)外有效,分子作用機(jī)制基本清楚,以及結(jié)構(gòu)簡單、分子量小、可改造空間大,可確定其為苗頭化合物,進(jìn)行系統(tǒng)的研究和開發(fā)。共設(shè)計、合成了200多個衍生物,通過體外活性評價,發(fā)現(xiàn)SP1610衍生物活性均未改善,研究終止;SP2343衍生物中C7體外活性明顯提高,但在體內(nèi)低劑量有效、高劑量有毒,而且它的成藥性較差,有待進(jìn)一步改善;長春胺系列僅合成了12個衍生物,發(fā)現(xiàn)體外活性顯著提高,其中V9的活性提高了50多倍,值得進(jìn)一步研究。阿爾茨海默癥(AD)是當(dāng)今公共健康的最大威脅之一,目前已上市藥物只能緩解癥狀,卻不能治愈。治療AD的有效藥物研發(fā)困難,主要因為其發(fā)病機(jī)制非常復(fù)雜,其中神經(jīng)纖維纏結(jié)、神經(jīng)細(xì)胞受損凋亡是導(dǎo)致病情加重的根本原因,因此,保護(hù)和恢復(fù)神經(jīng)細(xì)胞功能可能是治療AD病癥的重要突破口。本課題組針對神經(jīng)細(xì)胞凋亡的保護(hù),在STZ誘導(dǎo)的細(xì)胞模型上進(jìn)行高通量篩選,發(fā)現(xiàn)SP2343和SP1178不僅在細(xì)胞水平上表現(xiàn)出了保護(hù)活性,而且在ICV-STZ誘導(dǎo)的AD大鼠模型上也能夠修復(fù)受損的神經(jīng)細(xì)胞、減少Aβ的生成以及減輕Tau蛋白的異常磷酸化,從而改善了AD鼠的認(rèn)知功能障礙,同時,對SP2343的分子作用機(jī)制也進(jìn)行了深入研究。因此,可選其為值得進(jìn)行深入研究和開發(fā)的苗頭化合物。將已有的SP2343衍生物進(jìn)行體外活性評價及構(gòu)效關(guān)系研究,發(fā)現(xiàn)C50和C52的抗神經(jīng)細(xì)胞凋亡活性提高了近80倍,基于此又繼續(xù)合成50多個衍生物,但活性未進(jìn)一步改善;設(shè)計合成了10個SP1178衍生物,體外活性均未明顯改善,研究終止。本課題組前期選取蟾酥中抗腫瘤活性成分蟾毒靈為先導(dǎo)結(jié)構(gòu),通過結(jié)構(gòu)優(yōu)化得到了活性更強(qiáng)、毒性更低、成藥性更好的藥物候選物BF211。然而在狗上的毒性試驗中發(fā)現(xiàn)BF211仍有較強(qiáng)的心臟毒性,顯示無治療窗口。至今,酶激活式靶向前體藥物(enzyme-activated prodrug)在腫瘤治療方面已取得顯著的成果。其藥物設(shè)計的核心思路在于運(yùn)用特異性導(dǎo)向物修飾細(xì)胞毒性小分子,以屏蔽其活性基團(tuán),形成低活性的前藥;前藥進(jìn)入機(jī)體內(nèi)后以低活化形式被輸送至靶部位,繼而利用靶部位特異酶的選擇性酶解作用,使得活性母體藥物得以釋放。成纖維細(xì)胞是腫瘤微環(huán)境中主要的間質(zhì)細(xì)胞,與腫瘤細(xì)胞作用而被激活,成為腫瘤相關(guān)成纖維細(xì)胞(CAF),能高表達(dá)FAPα。FAPα屬絲氨酸蛋白酶類,在90%以上的上皮癌(包括肺癌、結(jié)腸癌、乳腺癌、膀胱癌、胰腺癌、卵巢癌等)的基質(zhì)活化成纖維細(xì)胞中表達(dá)。它具有促進(jìn)與調(diào)控腫瘤生長,促進(jìn)腫瘤的侵襲與轉(zhuǎn)移的作用。基于FAPα介導(dǎo)的酶激活式靶向抗腫瘤前藥策略已在阿霉素上成功應(yīng)用,我們設(shè)計、合成了14個FAPα激活式BF211前藥分子,經(jīng)過酶孵育、組織孵育以及細(xì)胞毒性試驗,發(fā)現(xiàn)前藥03是低毒高效的理想分子,隨后在大鼠結(jié)腸癌移植瘤模型進(jìn)行體內(nèi)活性評價,發(fā)現(xiàn)其抑瘤活性與原藥接近,且毒性明顯降低,表現(xiàn)出一定的治療窗口。該部分工作為以后更系統(tǒng)的FAPα激活式前藥研究及其應(yīng)用奠定了基礎(chǔ)。鈀催化的插羰反應(yīng)是合成羰基衍生物很常用的一類方法,如制備酰胺、酯、酸、醛、酮等。該類反應(yīng)過程中最關(guān)鍵的步驟是引入羰基,而CO高壓氣體則是最常用的羰基源,但CO是劇毒氣體,大量使用勢必會限制此方法的廣泛應(yīng)用。因此,本課題組設(shè)計了兩種新方法來定量釋放CO以滿足反應(yīng)需求,從而避免大量使用CO。方法一:是利用CHCl3-CsOH?H2O體系水解定量產(chǎn)生CO,在鈀催化下芳基碘代物與末端炔烴發(fā)生插羰偶聯(lián)反應(yīng)得到炔酮衍生物;方法二:是利用I2-PPh3體系配位HCOOH定量產(chǎn)生CO,在鈀催化下進(jìn)行芳基碘代物的插羰反應(yīng),再經(jīng)多余HCOOH還原后得到芳醛衍生物。這兩種方法具有顯著優(yōu)勢:(1)避免了直接使用高壓、高毒的CO氣體;(2)利用廉價易得的原料實現(xiàn)了CO的定量釋放以滿足插羰反應(yīng)需求;(3)反應(yīng)條件相對溫和、后處理簡單、產(chǎn)率較高;(4)反應(yīng)適用范圍廣泛,并在多種天然產(chǎn)物上成功應(yīng)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.
【學(xué)位授予單位】：中國科學(xué)院大學(xué)(中國科學(xué)院上海藥物研究所)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：R914
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本文編號：2063487