基于受体—配体空间构象信息设计IL-6拮抗多肽
发布时间:2019-06-07 14:49
【摘要】:自从1997年William等通过X射线晶体衍射分析确定了人IL-6的三维结构以来,人们在人IL-6及其受体拮抗剂的研究方面已经取得了一定成果,但因其特异性差,且不能安全、长期、有效地抑制体内IL-6的活性,所以限制了它们在临床上的应用。本研究拟根据IL-6/sIL-6R/gp130相互作用复合物的三维结构,通过距离几何学、分子间氢键形成理论探讨IL-6与其受体(IL-6R、gp130)的作用模式,借助虚拟筛选与计算机辅助合理设计相结合的方法,合理筛选获得IL-6小分子拮抗肽,初步建立基于受体—配体相互作用空间构象设计配体拮抗剂的可行性方案。同时,针对设计的IL-6小分子拮抗肽,从实验上对合成短肽的体外生物学活性进行了评价,希望为临床治疗IL-6相关疾病提供更有效的先导化合物。研究内容具体如下: 一.IL-6小分子拮抗肽的设计 根据IL-6/sIL-6R/gp130相互作用复合物的三维结构,通过计算机图形学技术、距离几何学、分子间氢键作用确定hIL-6/hIL-6R/gp130相互作用的功能域,理论分析影响配基—受体作用的功能域结构信息及其作用模式。利用位点连接法计算机辅助设计hIL-6拮抗肽,从头搭建拮抗肽空间构象,经分子力学、动力学优化获得的拮抗肽稳定构象与受体IL-6R进行分子对接,从理论上构建拮抗肽/hIL-6R相互作用复合物空间结构并对IL-6拮抗肽进行生物学功能评价。 二、IL-6小分子拮抗肽生物学功能评价 ①通过MTT法、~3H—TdR掺入法初步验证IL-6小分子拮抗肽的活性。选择IL-6依赖型细胞系—人多发性骨髓瘤细胞XG-7通过细胞学方法检测拮抗肽对其增殖的影响,进而评价其对IL-6活性的影响。结果表明,设计的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.
【学位授予单位】:中国人民解放军军事医学科学院
【学位级别】:硕士
【学位授予年份】:2005
【分类号】:R392
本文编号:2494878
[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.
【学位授予单位】:中国人民解放军军事医学科学院
【学位级别】:硕士
【学位授予年份】:2005
【分类号】:R392
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相关期刊论文 前3条
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,本文编号:2494878
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