Identification of TrkB Binders from Complex Matrices Using a Magnetic Drug Screening Nanoplatform
Identification of TrkB Binders from Complex Matrices Using a Magnetic Drug Screening Nanoplatform
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DOI：
10.1021/acsabm.1c00552
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发表时间：
2021-07-26
影响因子：
4.7
通讯作者：
Bao，Yuping
中科院分区：
文献类型：
--
作者：
Arituluk，Zekiye Ceren;Horne，Jesse;Bao，Yuping
脑源性神经营养因子(BDNF)及其受体酪氨酸受体激酶B(TrkB)在阿尔茨海默病等多种神经系统疾病中发挥重要作用。识别与TrkB相互作用的生物活性化合物可作为一种药物发现策略，以确定神经疾病的药物先导。在这里，我们报告了功能TrkB在磁性氧化铁纳米簇上的有效固定化，其中TrkB受体充当“智能诱饵”从混合物中结合化合物，而磁性纳米簇通过磁分离实现快速分离。特异性抗体标记证实了固定化TrkB的存在。随后，使用已知的TrkB激动剂，通过ATP/ADP转化实验评估了TrkB在氧化铁纳米簇上的活性。固定化的TrkB受体可以有效地从含有已知粘合剂的混合物、合成小分子混合物和Gotu Kola(积雪草植物提取物)中识别粘合剂。用四极飞行时间质谱仪与超高效液相色谱系统联用对鉴定出的化合物进行分析。重要的是，从Gotu Kola植物提取物中识别出的一些TrkB结合剂与之前在Gotu Kola提取物中观察到的神经保护作用的化合物相匹配，该化合物已被批准用于临床试验。我们的研究表明，Gotu Kola植物提取物治疗痴呆症的可能疗效，至少部分与与TrkB相互作用的化合物有关。这种方法的独特之处在于，它能够使用较少探索的跨膜靶点来快速筛选潜在的药物线索。这个平台在药物发现管道的早期阶段起到了药物筛选漏斗的作用。因此，我们的方法不仅将极大地有利于以跨膜蛋白为靶点的药物发现过程，而且还将允许评估和验证药物先导的靶向细胞通路。
Brain-derived neurotrophic factor (BDNF) and its receptor tyrosine receptor kinase B (TrkB) have been shown to play an important role in numerous neurological disorders, such as Alzheimer’s disease. The identification of biologically active compounds interacting with TrkB serves as a drug discovery strategy to identify drug leads for neurological disorders. Here, we report effective immobilization of functional TrkB on magnetic iron oxide nanoclusters, where TrkB receptors behave as “smart baits” to bind compounds from mixtures and magnetic nanoclusters enable rapid isolation through magnetic separation. The presence of the immobilized TrkB was confirmed by specific antibody labeling. Subsequently, the activity of the TrkB on iron oxide nanoclusters was evaluated with ATP/ADP conversion experiments using a known TrkB agonist. The immobilized TrkB receptors can effectively identify binders from mixtures containing known binders, synthetic small molecule mixtures, and Gotu Kola (Centella asiatica) plant extracts. The identified compounds were analyzed by an ultrahigh-performance liquid chromatography system coupled with a quadrupole time-of-flight mass spectrometer. Importantly, some of the identified TrkB binders from Gotu Kola plant extracts matched with compounds previously linked to neuroprotective effects observed for a Gotu Kola extract approved for use in a clinical trial. Our studies suggest that the possible therapeutic effects of the Gotu Kola plant extract in dementia treatment, at least partially, might be associated with compounds interacting with TrkB. The unique feature of this approach is its ability to fast screen potential drug leads using less explored transmembrane targets. This platform works as a drug-screening funnel at early stages of the drug discovery pipeline. Therefore, our approach will not only greatly benefit drug discovery processes using transmembrane proteins as targets but also allow for evaluation and validation of cellular pathways targeted by drug leads.