Abdel-Rahman Somaya A, Brogi Simone, Gabr Moustafa T
Department of Radiology, Molecular Imaging Innovations Institute (MI3), Weill Cornell Medicine New York NY 10065 USA
Department of Medicinal Chemistry, Faculty of Pharmacy, Mansoura University Mansoura 35516 Egypt.
RSC Adv. 2024 Jan 18;14(5):2918-2928. doi: 10.1039/d3ra08086b. eCollection 2024 Jan 17.
Retinoic acid receptor-related orphan receptor γt (RORγt) is a nuclear receptor found in various tissues that plays a crucial role in the differentiation and proliferation of T helper 17 (Th17) cells, as well as in their generation of the pro-inflammatory cytokine IL-17A. RORγt represents a promising therapeutic target for autoimmune diseases, metabolic disorders, and multiple tumors. Despite extensive research efforts focused on the development of small molecule RORγt modulators, no drug candidates have advanced to phase 3 clinical trials owing to a lack of efficacy or safety margin. This outcome highlights the unmet need to optimize small molecule drug candidates targeting RORγt to develop effective therapies for autoimmune and inflammatory diseases. In this study, we synthesized and evaluated 3-oxo-lithocholic acid amidates as a new class of RORγt modulators. Our evaluation entailed biophysical screening, cellular screening in different platforms, molecular docking, and pharmacokinetic profiling. The top compound from our study (3-oxo-lithocholic acid amidate, A2) binds to RORγt at an equilibrium dissociation constant () of 16.5 ± 1.34 nM based on microscale thermophoresis (MST). Assessment of the efficacy of A2 in the cellular RORγt reporter luciferase assay revealed a half-maximal inhibitory concentration (IC) value of 225 ± 10.4 nM. Unlike 3-oxo-lithocholic acid, A2 demonstrated the ability to reduce the IL-17A mRNA expression levels in EL4 cells with RORγt expression using quantitative reverse transcriptase PCR (RT-PCR). Validation of the desirable physicochemical properties and stability of A2 sets the stage for the preclinical evaluation of this new class of RORγt modulators in animal models of autoimmune diseases.
视黄酸受体相关孤儿受体γt（RORγt）是一种在多种组织中发现的核受体，它在辅助性T细胞17（Th17）细胞的分化和增殖以及促炎细胞因子IL-17A的产生中起着关键作用。RORγt是自身免疫性疾病、代谢紊乱和多种肿瘤的一个有前景的治疗靶点。尽管针对小分子RORγt调节剂的开发进行了广泛的研究，但由于缺乏疗效或安全边际，尚无候选药物进入3期临床试验。这一结果凸显了优化靶向RORγt的小分子候选药物以开发自身免疫性和炎性疾病有效疗法的未满足需求。在本研究中，我们合成并评估了3-氧代石胆酸酰胺作为一类新型RORγt调节剂。我们的评估包括生物物理筛选、不同平台的细胞筛选、分子对接和药代动力学分析。基于微量热泳动（MST），我们研究中的顶级化合物（3-氧代石胆酸酰胺，A2）以16.5±1.34 nM的平衡解离常数（）与RORγt结合。在细胞RORγt报告荧光素酶测定中对A2疗效的评估显示，半数最大抑制浓度（IC）值为225±10.4 nM。与3-氧代石胆酸不同，A2使用定量逆转录聚合酶链反应（RT-PCR）证明能够降低具有RORγt表达的EL4细胞中IL-17A mRNA的表达水平。A2理想的物理化学性质和稳定性的验证为在自身免疫性疾病动物模型中对这类新型RORγt调节剂进行临床前评估奠定了基础。
