【摘要】：自從1997年William等通過X射線晶體衍射分析確定了人IL-6的三維結構以來,人們在人IL-6及其受體拮抗劑的研究方面已經(jīng)取得了一定成果,但因其特異性差,且不能安全、長期、有效地抑制體內IL-6的活性,所以限制了它們在臨床上的應用。本研究擬根據(jù)IL-6/sIL-6R/gp130相互作用復合物的三維結構,通過距離幾何學、分子間氫鍵形成理論探討IL-6與其受體(IL-6R、gp130)的作用模式,借助虛擬篩選與計算機輔助合理設計相結合的方法,合理篩選獲得IL-6小分子拮抗肽,初步建立基于受體—配體相互作用空間構象設計配體拮抗劑的可行性方案。同時,針對設計的IL-6小分子拮抗肽,從實驗上對合成短肽的體外生物學活性進行了評價,希望為臨床治療IL-6相關疾病提供更有效的先導化合物。研究內容具體如下: 一.IL-6小分子拮抗肽的設計 根據(jù)IL-6/sIL-6R/gp130相互作用復合物的三維結構,通過計算機圖形學技術、距離幾何學、分子間氫鍵作用確定hIL-6/hIL-6R/gp130相互作用的功能域,理論分析影響配基—受體作用的功能域結構信息及其作用模式。利用位點連接法計算機輔助設計hIL-6拮抗肽,從頭搭建拮抗肽空間構象,經(jīng)分子力學、動力學優(yōu)化獲得的拮抗肽穩(wěn)定構象與受體IL-6R進行分子對接,從理論上構建拮抗肽/hIL-6R相互作用復合物空間結構并對IL-6拮抗肽進行生物學功能評價。 二、IL-6小分子拮抗肽生物學功能評價 ①通過MTT法、~3H—TdR摻入法初步驗證IL-6小分子拮抗肽的活性。選擇IL-6依賴型細胞系—人多發(fā)性骨髓瘤細胞XG-7通過細胞學方法檢測拮抗肽對其增殖的影響,進而評價其對IL-6活性的影響。結果表明,設計的IL-6小分子拮
[Abstract]:Since William et al determined the three-dimensional structure of human IL-6 by X-ray crystal diffraction analysis in 1997, some achievements have been made in the study of human IL-6 and its receptor antagonists, but it is not safe because of its poor specificity. Long-term, effectively inhibit the activity of IL-6 in vivo, so their clinical application is limited. In this study, according to the three-dimensional structure of IL-6/sIL-6R/gp130 interaction complex, the interaction mode between IL-6 and its receptor (IL-6R,gp130) was discussed by distance geometry and intermolecular hydrogen bond formation theory. By means of virtual screening and computer aided reasonable design, IL-6 small molecule antagonistic peptides were reasonably screened, and a feasible scheme for designing ligand antagonists based on receptor-ligand interaction space conformational design was established. At the same time, the biological activity of the synthesized short peptide was evaluated in vitro for the designed IL-6 small molecule antagonistic peptide, in the hope of providing more effective lead compounds for the clinical treatment of IL-6 related diseases. The research contents are as follows: 1. The design of IL-6 small molecule antagonistic peptides is based on the three-dimensional structure of IL-6/sIL-6R/gp130 interaction complex. The functional domain of hIL-6/hIL-6R/gp130 interaction was determined by computer graphics, distance geometry and intermolecular hydrogen bond interaction. The functional domain structure information and its action mode affecting ligand-receptor interaction were analyzed theoretically. HIL-6 antagonistic peptides were designed by computer aided design with site connection method. The spatial conformations of antagonistic peptides were constructed from scratch. The stable conformations of antagonistic peptides optimized by molecular mechanics and kinetics were linked to the receptor IL-6R. The spatial structure of antagonistic peptide / hIL-6R interaction complex was constructed theoretically and the biological function of IL-6 antagonistic peptide was evaluated. 2. Evaluation of biological function of IL-6 small molecule antagonistic peptide 1 the activity of IL-6 small molecule antagonistic peptide was preliminarily verified by MTT method and ~ 3H-TdR incorporation method. Human multiple myeloma cell line XG-7, a IL-6 dependent cell line, was selected to detect the effect of antagonistic peptides on its proliferation by Cytology, and then to evaluate the effect of antagonistic peptides on IL-6 activity. The results show that the designed IL-6 small molecules are antagonistic.
【學位授予單位】：中國人民解放軍軍事醫(yī)學科學院
【學位級別】：碩士
【學位授予年份】：2005
【分類號】：R392

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