Design and Application of Conjugatable Small Molecule Toll-Like Receptor 4 Ligands.

IF 4 2区 化学 Q1 BIOCHEMICAL RESEARCH METHODS
Yutaro Mahara, Issa Fukuda, Ryuho Tanaka, Shin-Ya Oyama, Hiroyuki Shinchi, Yasuo Suda, Nikunj M Shukla, Michael Chan, Tomoko Hayashi, Howard B Cottam, Dennis A Carson, Masahiro Wakao
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引用次数: 0

Abstract

We identified structurally distinct pyrimido[5,4-b]indole derivatives as toll-like receptor 4 (TLR4) ligands. Previous structure-activity relationship studies revealed that C8-aryl derivatives in pyrimido[5,4-b]indole, especially phenyl and 2-naphthyl compounds, are more potent in the activation of TLR4 signaling. Molecular modeling of these compounds indicated that C8-aryl groups are important for the interaction of the TLR4/myeloid differentiation factor-2 (MD-2) complex. Additionally, the modeling suggested that the N5 position in pyrimido[5,4-b]indole could be used as a further modification site to develop various drug conjugates. In this study, we examined whether the N5 position in pyrimido[5,4-b]indole can be used for conjugation without losing potency. Since tetraethylene glycol (TEG) derivatives at the N5 position were predicted to bind to TLR4/MD-2 complex using in silico molecular docking analysis, the compounds with the TEG group at the N5 position were synthesized and evaluated for immunostimulatory activity by human TLR4 reporter cell assay. As a result of fine-tuning of the C8 substitution groups, we found that TLR4 ligand (TLR4L) 10 with a 3-thienylethynyl group at the C8 position maintained TLR4 potency and demonstrated agonistic activity in primary murine bone marrow dendritic cells (mBMDC) and human TLR4 reporter HEK-Blue cells. TLR4L 10 was conjugated to sugar-immobilized gold nanoparticles (SGNPs) by introducing thioctic acid as a spacer into the TEG moiety. The obtained TLR4L-SGNPs 17 were taken up and showed agonistic activity in mBMDC. Thus, our designed TLR4L 10 and TLR4L-SGNP 17 are new candidates as immunomodulators for novel class adjuvant systems.

共轭小分子toll样受体4配体的设计与应用。
我们确定了结构上不同的嘧啶[5,4-b]吲哚衍生物作为toll样受体4 (TLR4)配体。以往的构效关系研究表明,嘧啶[5,4-b]吲哚中的c8 -芳基衍生物,尤其是苯基和2-萘基化合物,对TLR4信号的激活作用更强。这些化合物的分子模型表明,c8 -芳基对TLR4/髓样分化因子-2 (MD-2)复合物的相互作用很重要。此外,该模型表明,嘧啶[5,4-b]吲哚中的N5位置可以作为进一步修饰位点来开发各种药物偶联物。在本研究中,我们检测了嘧啶[5,4-b]吲哚的N5位置是否可以用于偶联而不失去效价。由于利用硅分子对接分析预测了N5位置的四乙二醇(TEG)衍生物与TLR4/MD-2复合物的结合,因此合成了具有N5位置TEG基团的化合物,并通过人TLR4报告细胞试验评估了其免疫刺激活性。通过对C8取代基的微调,我们发现在C8位置有一个3- thieny乙基基团的TLR4配体(TLR4L) 10在原代小鼠骨髓树突状细胞(mBMDC)和人TLR4报告细胞HEK-Blue细胞中保持了TLR4的效力,并表现出了拮抗活性。通过将硫辛酸作为间隔剂引入TEG片段,将tlr4l10偶联到糖固定化金纳米颗粒(SGNPs)上。获得的TLR4L-SGNPs 17在mBMDC中表现出激动作用。因此,我们设计的tlr4l10和TLR4L- sgnp 17是新型佐剂系统免疫调节剂的新候选分子。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Bioconjugate Chemistry
Bioconjugate Chemistry 生物-化学综合
CiteScore
9.00
自引率
2.10%
发文量
236
审稿时长
1.4 months
期刊介绍: Bioconjugate Chemistry invites original contributions on all research at the interface between man-made and biological materials. The mission of the journal is to communicate to advances in fields including therapeutic delivery, imaging, bionanotechnology, and synthetic biology. Bioconjugate Chemistry is intended to provide a forum for presentation of research relevant to all aspects of bioconjugates, including the preparation, properties and applications of biomolecular conjugates.
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