本文選題：腦血管痙攣　切入點(diǎn)：蛛網(wǎng)膜下腔出血　出處：《華東理工大學(xué)》2014年碩士論文

【摘要】：血管痙攣是心腦血管疾病重要的病理生理機(jī)制之一,開(kāi)發(fā)針對(duì)新靶標(biāo)和具有新作用機(jī)制的解除血管痙攣的治療藥物具有重要的科學(xué)意義。RhoA/ROCK系統(tǒng)是細(xì)胞收縮不依賴于細(xì)胞內(nèi)鈣濃度變化的重要信號(hào)傳導(dǎo)途徑,該通路相關(guān)蛋白的高表達(dá)或過(guò)度激活與血管痙攣的發(fā)生有著重要的關(guān)系,因此RhoA蛋白可能是一個(gè)潛在的藥物作用新靶點(diǎn)�？紤]到目前國(guó)際國(guó)內(nèi)有關(guān)直接針對(duì)RhoA蛋白GTP結(jié)合域的抑制劑研究報(bào)道很少的狀況,在課題組前期發(fā)現(xiàn)的第一代RhoA小分子抑制劑結(jié)構(gòu)和藥效團(tuán)信息基礎(chǔ)上,本論文開(kāi)展了第二代RhoA小分子抑制劑的設(shè)計(jì)、合成、體外內(nèi)藥理研究�；趹�(yīng)用分子對(duì)接技術(shù)模擬出的RhoA與第一代抑制劑的相互作用特征,本論文確定了第二代小分子RhoA抑制劑的設(shè)計(jì)思路,共設(shè)計(jì)合成34個(gè)目標(biāo)物(A01-A34)。對(duì)所有34個(gè)目標(biāo)物開(kāi)展了細(xì)胞水平的RhoA抑制活性研究,成功發(fā)現(xiàn)了6個(gè)目標(biāo)物(A01、A02、A07、A12、 A14)為強(qiáng)效RhoA抑制劑,它們的IC50值處于1～2μM之間,可以定義為第二代RhoA小分子抑制劑。進(jìn)一步測(cè)試了6個(gè)RhoA抑制劑對(duì)血管張力的影響,發(fā)現(xiàn)2個(gè)目標(biāo)物對(duì)苯腎上腺素預(yù)制血管收縮有明顯的舒張效果(A02:IC50=70.9±1.3μM; A14: IC50=73.4±2.6κM),其舒張血管效果相比第一代RhoA的抑制提高了大約一倍左右。通過(guò)考察A02、A10和A14這3個(gè)目標(biāo)物在水中的溶解性,我們最終選擇水溶性良好(S=639μg/mL)的目標(biāo)物A02開(kāi)展初步的體內(nèi)藥效研究。在蛛網(wǎng)膜下腔出血后腦血管痙攣大鼠模型中,第二代RhoA抑制劑A02對(duì)SD大鼠SAH-CVS有明顯的緩解作用,在低劑量組(95mM)表現(xiàn)出與臨床藥物法舒地爾(15.2mM)相當(dāng)?shù)乃幚硇Ч８鶕?jù)細(xì)胞、組織和動(dòng)物三種水平藥理活性結(jié)果,我們總結(jié)歸納了本論文第二代RhoA抑制劑的構(gòu)效關(guān)系。這些SARs結(jié)果為今后尋找小分子RhoA抑制劑提供了結(jié)構(gòu)線索。同時(shí),本論文發(fā)現(xiàn)的強(qiáng)效第二代RhoA小分子抑制劑A02可以作為具有自主知識(shí)產(chǎn)權(quán)的新型腦血管痙攣治療臨床前候選藥物進(jìn)行下一步的開(kāi)發(fā)研究。
[Abstract]:Vasospasm is one of the important pathophysiological mechanisms of cardiovascular and cerebrovascular diseases. It is of great scientific significance to develop new targets and new mechanisms for the treatment of vasospasm. RhoA / rock system is an important signal transduction pathway for cell contraction independent of changes in intracellular calcium concentration. The overexpression or overexpression of proteins associated with this pathway is associated with the development of vasospasm. Therefore, RhoA protein may be a potential new target for drug action. Considering that there are few reports on inhibitors directly targeting the GTP binding domain of RhoA protein at home and abroad, Based on the information of the structure and pharmacophore of the first generation of RhoA small molecule inhibitors, the design and synthesis of the second generation of RhoA small molecule inhibitors were carried out in this paper. In vitro pharmacological studies. Based on the interaction characteristics of RhoA and first-generation inhibitors simulated by molecular docking technique, the design idea of the second generation of small molecular RhoA inhibitors was determined in this paper. A total of 34 target compounds (A01-A34) were designed and synthesized. Cell-level RhoA inhibitory activity of all 34 targets was studied. It was successfully found that 6 target compounds (A01A02A02A07A12, A14) were potent RhoA inhibitors, and their IC50 values were in the range of 1 渭 M. It can be defined as the second generation of RhoA small molecule inhibitors. The effects of 6 RhoA inhibitors on vascular tension were further tested. It was found that the vasodilation effects of the two target compounds on the prefabricated vasoconstriction of phenylephrine were 70.9 鹵1.3 渭 M and A14: IC50=73.4 鹵2.6 渭 M, respectively. The inhibition of the vasodilation effect of the two target compounds was about twice as high as that of the first generation of RhoA. The solubility of A02: A10 and A14 in water was investigated. We finally selected A02, a water-soluble target of 639 渭 g 路mL ~ (-1), to study its pharmacodynamics in vivo. In the model of cerebral vasospasm after subarachnoid hemorrhage, A02, a second-generation inhibitor of RhoA, could significantly relieve SAH-CVS in SD rats. In the low dose group (95 mm), the pharmacological effect was comparable to that of the clinical drug fasudil 15.2mM.According to the results of three levels of pharmacological activity of cells, tissues and animals, We summarize the structure-activity relationships of the second generation of RhoA inhibitors in this thesis. These SARs results provide a structural clue for finding small molecular RhoA inhibitors in the future. A02, a potent second generation small molecule inhibitor of RhoA, can be used as a new pre-clinical candidate for cerebral vasospasm therapy with intellectual property rights.
【學(xué)位授予單位】：華東理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R965

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相關(guān)博士學(xué)位論文 前1條

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本文編號(hào)：1666483