高通量CETSA鉴定了在人类ipsc衍生的阿尔茨海默病小胶质细胞中具有功能活性的ILT3 （LILRB4）小分子调节剂

IF 3.9 3区医学 Q2 CHEMISTRY, MEDICINAL

ACS Medicinal Chemistry Letters Pub Date : 2026-07-09 Epub Date: 2026-06-18 DOI:10.1021/acsmedchemlett.6c00244

Somaya A. Abdel-Rahman, and , Moustafa Gabr*,

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引用次数: 0

摘要

小胶质细胞中的免疫抑制信号有助于阿尔茨海默病（AD）中淀粉样蛋白-β (Aβ)清除受损和神经炎症，然而靶向这些途径的小分子调节剂在很大程度上仍未被探索。在这里，我们报告了一种高通量细胞热移测定（HT-CETSA）平台的开发，用于鉴定靶向抑制性免疫受体ILT3 （LILRB4）的小分子结合物。筛选了约40 000个化合物，产生了多个经过验证的命中，包括IB15C，一种亚微摩尔的ILT3结合剂，通过初步的结构-活性关系优化鉴定。通过微尺度热电泳、表面等离子体共振、对接和定点突变的正交验证证实了直接和靶向特异性的ILT3作用。在功能上，IB15C破坏了ILT3-ApoE的相互作用，恢复了人ipsc源性小胶质细胞的小胶质活性，降低了SHP1/2和NF-κB信号，抑制了IL-1β的分泌，增强了Aβ的摄取。IB15C还显示出良好的体外药代动力学和安全性，支持进一步开发靶向ilt3的神经免疫疗法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

High-Throughput CETSA Identifies Small Molecule Modulators of ILT3 (LILRB4) with Functional Activity in Human iPSC-Derived Microglia for Alzheimer’s Disease

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High-Throughput CETSA Identifies Small Molecule Modulators of ILT3 (LILRB4) with Functional Activity in Human iPSC-Derived Microglia for Alzheimer’s Disease

Immune inhibitory signaling in microglia contributes to impaired amyloid-β (Aβ) clearance and neuroinflammation in Alzheimer’s disease (AD), yet small molecule modulators targeting these pathways remain largely unexplored. Here, we report the development of a high-throughput cellular thermal shift assay (HT-CETSA) platform for identification of small molecule binders targeting the inhibitory immune receptor ILT3 (LILRB4). Screening of ∼40 000 compounds yielded multiple validated hits, including IB15C, a submicromolar ILT3 binder identified through preliminary structure–activity relationship optimization. Orthogonal validation by microscale thermophoresis, surface plasmon resonance, docking, and site-directed mutagenesis confirmed direct and target-specific ILT3 engagement. Functionally, IB15C disrupted the ILT3-ApoE interaction and restored microglial activity in human iPSC-derived microglia, reducing SHP1/2 and NF-κB signaling, suppressing IL-1β secretion, and enhancing Aβ uptake. IB15C also demonstrated favorable in vitro pharmacokinetic and safety properties, supporting further development of ILT3-targeted neuroimmune therapeutics.

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来源期刊

ACS Medicinal Chemistry Letters CHEMISTRY, MEDICINAL-

CiteScore

7.30

自引率

2.40%

发文量

328

审稿时长

1 months

期刊介绍： ACS Medicinal Chemistry Letters is interested in receiving manuscripts that discuss various aspects of medicinal chemistry. The journal will publish studies that pertain to a broad range of subject matter, including compound design and optimization, biological evaluation, drug delivery, imaging agents, and pharmacology of both small and large bioactive molecules. Specific areas include but are not limited to: Identification, synthesis, and optimization of lead biologically active molecules and drugs (small molecules and biologics) Biological characterization of new molecular entities in the context of drug discovery Computational, cheminformatics, and structural studies for the identification or SAR analysis of bioactive molecules, ligands and their targets, etc. Novel and improved methodologies, including radiation biochemistry, with broad application to medicinal chemistry Discovery technologies for biologically active molecules from both synthetic and natural (plant and other) sources Pharmacokinetic/pharmacodynamic studies that address mechanisms underlying drug disposition and response Pharmacogenetic and pharmacogenomic studies used to enhance drug design and the translation of medicinal chemistry into the clinic Mechanistic drug metabolism and regulation of metabolic enzyme gene expression Chemistry patents relevant to the medicinal chemistry field.