DataSheet_1_A Novel Glucocorticoid and Androgen Receptor Modulator Reduces Viral Entry and Innate Immune Inflammatory Responses in the Syrian Hamster Model of SARS-CoV-2 Infection.docx

Despite significant research efforts, treatment options for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) remain limited. This is due in part to a lack of therapeutics that increase host defense to the virus. Replication of SARS-CoV-2 in lung tissue is associated with marked infiltration of macrophages and activation of innate immune inflammatory responses that amplify tissue injury. Antagonists of the androgen (AR) and glucocorticoid (GR) receptors have shown efficacy in models of COVID-19 and in clinical studies because the cell surface proteins required for viral entry, angiotensin converting enzyme 2 (ACE2) and the transmembrane protease, serine 2 (TMPRSS2), are transcriptionally regulated by these receptors. We postulated that the GR and AR modulator, PT150, would reduce infectivity of SARS-CoV-2 and prevent inflammatory lung injury in the Syrian golden hamster model of COVID-19 by down-regulating expression of critical genes regulated through these receptors. Animals were infected intranasally with 2.5 × 104 TCID50/ml equivalents of SARS-CoV-2 (strain 2019-nCoV/USA-WA1/2020) and PT150 was administered by oral gavage at 30 and 100 mg/Kg/day for a total of 7 days. Animals were examined at 3, 5 and 7 days post-infection (DPI) for lung histopathology, viral load and production of proteins regulating the progression of SARS-CoV-2 infection. Results indicated that oral administration of PT150 caused a dose-dependent decrease in replication of SARS-CoV-2 in lung, as well as in expression of ACE2 and TMPRSS2. Lung hypercellularity and infiltration of macrophages and CD4+ T-cells were dramatically decreased in PT150-treated animals, as was tissue damage and expression of IL-6. Molecular docking studies suggest that PT150 binds to the co-activator interface of the ligand-binding domain of both AR and GR, thereby acting as an allosteric modulator and transcriptional repressor of these receptors. Phylogenetic analysis of AR and GR revealed a high degree of sequence identity maintained across multiple species, including humans, suggesting that the mechanism of action and therapeutic efficacy observed in Syrian hamsters would likely be predictive of positive outcomes in patients. PT150 is therefore a strong candidate for further clinical development for the treatment of COVID-19 across variants of SARS-CoV-2.

尽管研究投入了大量努力，但对于严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的治疗选择仍显不足。部分原因在于缺乏能够增强宿主对病毒防御能力的治疗药物。SARS-CoV-2在肺组织的复制与显著的中性粒细胞浸润以及固有免疫炎症反应的激活有关，这些反应会加剧组织损伤。研究表明，雄激素受体（AR）和糖皮质激素受体（GR）的拮抗剂在COVID-19模型和临床试验中显示出疗效，因为病毒进入所必需的细胞表面蛋白——血管紧张素转换酶2（ACE2）和跨膜蛋白酶，丝氨酸2（TMPRSS2）——的转录受到这些受体的调控。我们假设，糖皮质激素和雄激素调节剂PT150可以通过下调通过这些受体调控的关键基因的表达，从而降低SARS-CoV-2的感染性并预防叙利亚金仓鼠COVID-19模型的炎症性肺损伤。动物通过鼻腔感染2.5 × 104 TCID50/ml等量的SARS-CoV-2（菌株2019-nCoV/USA-WA1/2020），并以30和100 mg/Kg/day的剂量通过口服灌胃给予PT150，连续7天。感染后第3、5和7天对动物进行检查，以评估肺组织病理学、病毒载量和调节SARS-CoV-2感染进展的蛋白质的产生。结果显示，PT150的口服给药导致SARS-CoV-2在肺中的复制以及ACE2和TMPRSS2的表达剂量依赖性地降低。与未处理动物相比，PT150处理动物肺组织的过度细胞增殖和巨噬细胞以及CD4+ T细胞的浸润显著减少，组织损伤和IL-6的表达也有所降低。分子对接研究表明，PT150结合到AR和GR配体结合域的共激活剂界面，因此作为这些受体的变构调节剂和转录抑制剂。AR和GR的系统发育分析显示，包括人类在内的多种物种间保持了高度序列一致性，这表明在叙利亚金仓鼠中观察到的药理作用和治疗效应对患者可能具有预测性。因此，PT150是治疗SARS-CoV-2变种的COVID-19的进一步临床开发的一个强有力的候选药物。