Enhanced immunostimulatory activity of in silico discovered agonists of Toll-like receptor 2 (TLR2)
Enhanced immunostimulatory activity of in silico discovered agonists of Toll-like receptor 2 (TLR2)
复制标题
DOI：
10.1016/j.bbagen.2017.07.011
复制
发表时间：
2017-11-01
影响因子：
3
通讯作者：
Santos-Sierra, S.
中科院分区：
文献类型：
--
作者：
Murgueitio, M. S.;Ebner, S.;Santos-Sierra, S.
背景：抗癌疫苗的紧急治疗使用toll样受体（TLRs）激动剂作为树突状细胞（DC）疫苗佐剂。从患者身上分离出DCs，体外用TLR激动剂和肿瘤抗原刺激，然后再输注回患者体内。尽管一些TLR配体已经在临床试验中进行了测试，但具有改进免疫调节特性的新型TLR激动剂对于优化治疗成功至关重要。我们报道了小分子TLR2激动剂的发现，作为合成佐剂具有良好的性能。方法：我们进行了形状和特征为基础的相似性虚拟筛选对市售化合物库。选择的虚拟命中在tlr2报告细胞中进行了实验测试，并表征了它们在吞噬细胞和dc中的活性。提出了化合物与TLR2的结合模型（对接研究）。结果：在HEK293-TLR2细胞荧光素酶报告基因检测中，通过对300万个化合物库的虚拟筛选方法，发现4个虚拟命中（AG1、AG2、AG3、AG4）可协同增强脂肽配体Pam(3)CSK(4)诱导的NF-kB活化。Biacore实验表明，AG1 AG4是TLR2和AG2结合TLR2的ago-变构调节剂，具有高亲和力（K-D 0.8 μ M）。这些化合物诱导人外周血单核细胞（PBMCs）产生tnf - α，并通过IL-12的产生和CD83/CD86的上调激活DCs。结论：通过硅/体外联合方法，我们发现了tlr2激动剂（AG1 AG4），可激活人和小鼠免疫细胞。一般意义：我们介绍了四种新的TLR2 ago-变构调节剂，它们可以刺激髓细胞活性，并有希望成为合成佐剂。
Background: Emergent therapies in anticancer vaccination use Toll-like receptors (TLRs) agonists as dendritic cell (DC) vaccine adjuvants. DCs from the patient are isolated, stimulated with TLR agonists and tumor antigens ex vivo and then infused back into the patient. Although some TLR ligands have been tested in clinical trials, novel TLR agonists with improved immunomodulatory properties are essential to optimize treatment success. We report on the discovery of small-molecule TLR2 agonists, with favorable properties as synthetic adjuvants.Methods: We performed a shape- and featured-based similarity virtual screening against a commercially available compound library. The selected virtual hits were experimentally tested in TLR2-reporter cells and their activity in phagocytes and DCs was characterized. A binding model of the compounds to TLR2 (docking studies) was proposed.Results: Through a virtual screening approach against a library of three million compounds four virtual hits (AG1, AG2, AG3, AG4) were found to synergistically augment the NF-kB activation induced by the lipopeptide ligand Pam(3)CSK(4) in luciferase reporter assays using HEK293-TLR2 cells. Biacore experiments indicated that AG1 AG4 are ago-allosteric modulators of TLR2 and AG2 bound TLR2 with high affinity (K-D 0.8 mu M). The compounds induced TNF-alpha production in human peripheral blood mononuclear cells (PBMCs) and they activated DCs as indicated by IL-12 production and upregulation of CD83/CD86.Conclusions: Following a combined in silico/in vitro approach we have discovered TLR2-agonists (AG1 AG4) that activate human and mouse immune cells.General significance: We introduce four novel TLR2 ago-allosteric modulators that stimulate myeloid cell activity and constitute promising candidates as synthetic adjuvants.