本文選題：血栓栓塞性疾病 + 血栓��； 參考：《山東大學(xué)》2017年碩士論文

【摘要】：心腦血管疾病是一類由于心臟或血管病變導(dǎo)致的循環(huán)系統(tǒng)功能紊亂的疾病的統(tǒng)稱,是世界范圍內(nèi)尤其是亞洲地區(qū)的首要死亡原因,對(duì)人類的生命健康構(gòu)成了嚴(yán)重威脅。其中血栓栓塞性疾病,如心肌梗死、腦卒中、深部靜脈血栓,已成為全世界致殘率和致死率最高的疾病之一,因此血栓的形成是引起心腦血管疾病的最主要因素。然而目前臨床上常用的抗凝血藥物如華法林等仍存在著諸多缺陷,例如給藥不方便、不可預(yù)測(cè)的抗凝效果、治療窗窄、能夠與許多食物和藥物發(fā)生相互作用等。因此,亟需開(kāi)發(fā)更安全有效的新型抗凝血藥物。Xa因子(FXa)位于人體內(nèi)源性凝血途徑和外源性凝血途徑的交匯處,如果能夠抑制FXa就可以同時(shí)阻斷兩條凝血途徑,因此,FXa目前已成為抗凝藥物研發(fā)中的一個(gè)十分具有吸引力的靶點(diǎn)。目前已經(jīng)有一些口服的小分子FXa抑制劑在市場(chǎng)上銷售,例如利伐沙班(rivaroxaban)、阿哌沙班(apixaban)以及依度沙班(edoxaban)。雖然新型FXa抑制劑克服了傳統(tǒng)抗凝藥物的缺點(diǎn),但是,這些藥物仍然存在一些缺陷,如藥物的適應(yīng)癥狹窄、損害肝腎功能、以及發(fā)生出血時(shí)缺乏有效的解毒劑等。鑒于抑制劑本身的有效性和安全性問(wèn)題,因此,亟需開(kāi)發(fā)更安全有效的新型FXa抑制劑供臨床應(yīng)用。本論文以利伐沙班為先導(dǎo)化合物,通過(guò)研究分析"利伐沙班-FXa復(fù)合體"的晶體結(jié)構(gòu),發(fā)現(xiàn)FXa擁有兩個(gè)最關(guān)鍵的結(jié)合口袋,分別是:特異性結(jié)合口袋S1和較大的疏水性口袋S4,同時(shí)兩口袋之間存在著大約80°的夾角;因此,利伐沙班分子也相應(yīng)的被分為三部分:P1區(qū)、P4區(qū)和中間的Linker區(qū)。FXa的特異性結(jié)合口袋S1為較小的疏水性口袋,正好可以容納利伐沙班的P1區(qū)5-氯噻吩基團(tuán),同時(shí)5-氯噻吩基團(tuán)與S1 口袋底部的Tyr228形成了 Cl-π作用力;而較大的疏水性口袋S4的氨基酸殘基與利伐沙班P4區(qū)的苯環(huán)形成了 π-π堆積作用;同時(shí),利伐沙班的Linker區(qū)與FXa的氨基酸Gly219形成了兩個(gè)氫鍵。經(jīng)過(guò)對(duì)先導(dǎo)化合物利伐沙班與FX a的結(jié)合模式的分析,我們基于電子等排、優(yōu)勢(shì)分子片段雜合等原理,將利伐沙班的P4區(qū)和Linker區(qū)進(jìn)行了結(jié)構(gòu)改造,如改變P4區(qū)的體積、疏水性和增加Linker區(qū)與FXa結(jié)合位點(diǎn)的作用力等,設(shè)計(jì)了吡咯烷酮類、噻二嗪類和吲哚環(huán)類三個(gè)系列FXa抑制劑,以期得到具有較高活性的新型骨架Xa因子抑制劑。同時(shí),我們運(yùn)用計(jì)算機(jī)輔助藥物設(shè)計(jì)軟件Sybyl X 1.3對(duì)所設(shè)計(jì)的三個(gè)系列化合物進(jìn)行了分子模擬對(duì)接研究,研究結(jié)果表明:通過(guò)結(jié)構(gòu)改造所設(shè)計(jì)的新化合物具有理論上的合理性。通過(guò)成環(huán)反應(yīng)、�；磻�(yīng)、縮合反應(yīng)等實(shí)驗(yàn)步驟,對(duì)三個(gè)系列的目標(biāo)化合物進(jìn)行了定向合成,共合成了吡咯烷酮類、噻二嗪類和吲哚環(huán)類三個(gè)系列37個(gè)Xa因子抑制劑,并經(jīng)光譜確證結(jié)構(gòu)。首先測(cè)試了所合成FXa抑制劑的抗凝血活性,由化合物對(duì)血漿凝血酶原時(shí)間的影響來(lái)評(píng)價(jià)。通過(guò)使用凝血酶原時(shí)間測(cè)定儀,測(cè)定了在化合物不同濃度下貧血小板血漿的凝血酶原時(shí)間,然后計(jì)算出"使凝血時(shí)間延長(zhǎng)一倍的化合物濃度"(PTCT2)。個(gè)別化合物具有一定的抗凝活性,與先導(dǎo)化合物利伐沙班(PTCT2 = 1.3μM)相比,仍存在差距。本論文進(jìn)而對(duì)抗凝活性較好的化合物進(jìn)行了 FXa抑制活性的測(cè)定。FXa能夠促使發(fā)色底物S-2222裂解成多肽和對(duì)硝基苯胺,對(duì)硝基苯胺在405 nm處有紫外吸收。同樣選取了rivaroxaban作為陽(yáng)性對(duì)照藥,由酶標(biāo)儀測(cè)定出目標(biāo)化合物在不同濃度下對(duì)吸光度的影響,并通過(guò)軟件計(jì)算出所設(shè)計(jì)化合物對(duì)FXa的抑制率及IC50值。測(cè)試結(jié)果顯示:個(gè)別化合物的活性較好,例如化合物H2、H14和H15,其IC50值分別為0.82、0.55和0.52μM,但與陽(yáng)性對(duì)照藥利伐沙班(IC50=5nM)尚存在差距�？傊�,基于靶標(biāo)的合理藥物設(shè)計(jì)及分子模擬,設(shè)計(jì)并通過(guò)定向合成得到了吡咯烷酮類、噻二嗉類和吲哚環(huán)類三個(gè)系列具有新型骨架的FXa抑制劑,盡管抗凝實(shí)驗(yàn)和酶活實(shí)驗(yàn)結(jié)果顯示所設(shè)計(jì)的目標(biāo)化合物活性未達(dá)到先導(dǎo)化合物利rivaroxaban的水平,但是通過(guò)研究分析構(gòu)效關(guān)系,對(duì)我們更深入的研究和設(shè)計(jì)FXa抑制劑具有指導(dǎo)意義。
[Abstract]:Cardiovascular and cerebrovascular disease is a general name for a disorder of the circulatory system caused by heart or vascular lesions. It is the leading cause of death worldwide, especially in Asia, and poses a serious threat to human life and health. Thromboembolic diseases such as myocardial infarction, stroke, and deep venous thrombosis have become a whole. The formation of thrombus is one of the most important diseases in the world, so the formation of thrombus is the most important factor causing cardiovascular and cerebrovascular diseases. However, there are still many defects such as Hua Falin, such as inconvenient, unpredictable anticoagulant fruit, narrow treatment window, and many foods and drugs. Therefore, it is urgent to develop a more safe and effective anticoagulant drug.Xa factor (FXa), which is located at the intersection of endogenous coagulation pathways and exogenous coagulation pathways. If it can inhibit FXa, two clotting pathways can be blocked simultaneously. Therefore, FXa has become an attractive attraction in the development of anticoagulant drugs. Target. At present, some oral small molecule FXa inhibitors have been sold on the market, such as Lev Shaaban (rivaroxaban), opito Shaaban (apixaban) and idegree Shaaban (edoxaban). Although the new FXa inhibitors overcome the shortcomings of traditional anticoagulants, these drugs still have some defects, such as the stenosis of drug indications, and the loss of the drugs. The function of liver and kidney and the lack of effective antidote when bleeding. In view of the effectiveness and safety of the inhibitor itself, it is urgent to develop a more safe and effective new FXa inhibitor for clinical application. In this paper, the crystal structure of Lev Shaaban -FXa complex was studied and analyzed by the study of the crystal structure of the lefore Shaaban complex, and the discovery of F Xa has two most critical combined pockets, which are specific binding pocket S1 and larger hydrophobic pocket S4, and there are about 80 degrees between the two pockets; therefore, the lev Shaaban molecule is correspondingly divided into three parts: the P1 region, the P4 region and the Linker region.FXa in the P4 region, the specific binding pocket S1 is a smaller hydrophobic pocket, The 5- chlorthiophene group in P1 region of lefal Shaaban was accommodated, and the 5- chlorthiophene group formed a Cl- PI force with the Tyr228 at the bottom of the S1 pocket; the larger hydrophobic pocket S4 residues formed a pion pion accumulation with the benzene ring of the lev Shaaban P4 region; meanwhile, the Linker region of Lev Shaaban and the Gly219 of the FXa were formed. Two hydrogen bonds. Through the analysis of the binding mode of lefore Shaaban and FX a, based on the principle of electron equal row and hetero heterozygosity, the structure of the P4 and Linker regions of Lev Shaaban was reconstructed, such as changing the volume, hydrophobicity of the P4 region, and increasing the force of the Linker region with the FXa binding site, and designed pyrrole. Three series FXa inhibitors of alkanones, thiazoles and indole rings are used to obtain a new type of Xa factor inhibitor with high activity. At the same time, we used the computer aided drug design software Sybyl X 1.3 to study the molecular simulation of the designed series of compounds. The new compound has theoretical rationality. Through the cyclic reaction, acylation reaction, condensation reaction and other experimental steps, three series of target compounds were synthesized. A total of three series of 37 Xa factor inhibitors, pyrroliones, thiazoles and indole rings, were synthesized and confirmed by spectral identification. The anticoagulant activity of the synthetic FXa inhibitor was evaluated by the effect of the compound on the plasma prothrombin time. By using the prothrombin time analyzer, the prothrombin time of the blood poor plasma in different concentrations of the compound was measured, and the "PTCT2" was calculated. There is a certain anticoagulant activity, compared with the pilot compound leavin Shaaban (PTCT2 = 1.3 M), there is still a gap. In this paper, the determination of the FXa inhibitory activity of the compounds with better anticoagulant activity.FXa can cause the chromophore S-2222 to break into polypeptides and p-nitroaniline, and the p-nitroaniline has UV absorption at 405 nm. Rivaroxaban was used as a positive control drug. The effect of the target compound on the absorbance at different concentrations was measured by the enzyme scale, and the inhibition rate and IC50 value of the designed compound to FXa were calculated by software. The test results showed that the activities of individual compounds were better, such as H2, H14 and H15, and their IC50 values were 0.82,0.55 and 0.52 u M, respectively. But there is still a gap with the positive control drug Lev Shaaban (IC50=5nM). In a word, based on the target's rational drug design and molecular simulation, three series of pyrroliones, thiazoles and indole rings, which have a new skeleton of FXa inhibitors, are designed and carried out by directional synthesis, although the anticoagulant experiment and the results of enzyme activity experiments show the design of the IC50=5nM. The activity of the target compound does not reach the level of the lead compound, rivaroxaban, but by studying the structure-activity relationship, it has a guiding significance for us to further study and design the FXa inhibitors.

【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R914;R96

【參考文獻(xiàn)】

相關(guān)期刊論文 前9條

1 姚天賜;許云祿;;溶血栓藥物研究進(jìn)展[J];海峽藥學(xué);2010年06期

2 馬培奇;;抗凝血藥物現(xiàn)狀及其研發(fā)動(dòng)態(tài)[J];上海醫(yī)藥;2009年08期

3 吳小平;劉放;;抗血小板聚集藥物研究進(jìn)展[J];西北藥學(xué)雜志;2009年04期

4 韓邦志;謝金鮮;;中藥止血與凝血機(jī)制的研究進(jìn)展[J];廣西中醫(yī)藥;2009年02期

5 武建卓;吉愛(ài)國(guó);;抗凝血的低分子量肽類似物[J];生命的化學(xué);2008年04期

6 史旭波;胡大一;;血栓形成與凝血機(jī)制及調(diào)節(jié)[J];臨床薈萃;2007年14期

7 吳俊華;;凝血酶抑制劑研究進(jìn)展[J];生命科學(xué)儀器;2006年05期

8 胡大一,孫藝紅;低分子肝素與血栓栓塞性疾病[J];中華心血管病雜志;2005年02期

9 韓玲軍,李青山;藥物化學(xué)研究的新進(jìn)展[J];山西醫(yī)科大學(xué)學(xué)報(bào);2004年04期

相關(guān)碩士學(xué)位論文 前1條

1 葉永勝;Xa因子抑制劑對(duì)關(guān)節(jié)置換術(shù)后深靜脈血栓形成預(yù)防的臨床研究[D];暨南大學(xué);2011年

，

本文編號(hào)：1880376