Effect of a semi-synthetic oxysterol drug candidate, Oxy210, on atherosclerosis in a mouse model of NASH

摘要 心血管疾病（CVD）仍然是美国和世界范围内的主要死亡原因。的 绝大多数CVD相关死亡是动脉粥样硬化及其心血管并发症的直接结果。 动脉粥样硬化是一种由低密度脂蛋白胆固醇（LDL-C）引发的慢性炎症性疾病， 在动脉壁中的积累和氧化，以及巨噬细胞的募集，巨噬细胞在它们形成泡沫细胞时， 试图去除脂蛋白氧化产物，但在离开血管壁之前在此过程中死亡。尽管 动脉粥样硬化治疗的最新进展，包括降脂药物的使用， 随着生活方式的改变，CVC的死亡率预计将从2018年的1700多万增加到2019年的1700多万。 到2030年全球将达到2400万。近年来，除了动脉粥样硬化的经典危险因素外，即 糖尿病、高胆固醇血症、高血压与吸烟、非酒精性脂肪 肝脏疾病（NAFLD）和动脉粥样硬化已经被报道。NAFLD患者死于CVD更多 比肝病和NAFLD更严重的非酒精性脂肪性肝炎更常见 （NASH），现在被认为是动脉粥样硬化发展的独立且可能的因果风险因素。 因此，已经提出治疗NAFLD的疗法也可以抑制动脉粥样硬化形成，特别是因为 这些疾病具有共同的潜在因素，包括炎症、脂质沉积和氧化， 功能失调的巨噬细胞在MAX BioPharma，我们已经确定了一种领先的专有半合成氧甾醇， Oxy 210，其在人源化小鼠模型ApoE*3-Leiden.CETP中显著抑制NASH，如通过 减少肝脏脂质沉积、炎症和纤维化以及减少炎性细胞因子， 流通Oxy 210是口服生物可利用的，具有良好的药代动力学和安全性特征， 可扩展的在初步研究中，我们发现Oxy 210在巨噬细胞中具有抗炎作用 通过抑制Toll样受体，用脂多糖或致动脉粥样硬化的磷脂PGPC治疗 发信号。我们假设，考虑到Oxy 210对NASH和炎症的抑制作用， 作为NASH和动脉粥样硬化的常见介质，Oxy 210也可能具有抗动脉粥样硬化特性。在 本申请我们提出通过在3个具体实施方案中概述研究在体外和体内测试该假设 目的：1）检测Oxy 210对HSC、Kupffer细胞和血管内皮细胞活化的影响 体外用LPS或合成的TLR 2激动剂处理的细胞; 2）作用的分子机制的检查 3）评估Oxy 210在体外的抗炎作用;和 在ApoE*3-Leiden.CETP转基因NASH人源化小鼠模型中的动脉粥样硬化形成。Findings from the 拟议的研究将证明Oxy 210作为一类治疗性候选药物的潜力， 用于治疗NASH和CVD。

ABSTRACT Cardiovascular disease (CVD) remains the leading cause of death in the United States and worldwide. The large majority of CVD-related deaths are the direct result of atherosclerosis and its cardiovascular complications. Atherosclerosis is a chronic inflammatory disease triggered by low density lipoprotein cholesterol (LDL-C), its accumulation and oxidation in the artery wall, and recruitment of macrophages that become foam cells as they try to remove the lipoprotein oxidation products but die in the process before exiting the vascular wall. Despite recent improvements in the management of atherosclerosis, including the use of lipid lowering drugs and promotion of lifestyle changes, the mortality from CVC is expected to increase from over 17 million in 2018 to almost 24 million by 2030 globally. In recent years, in addition to classic risk factors for atherosclerosis, namely diabetes, hypercholesterolemia, hypertension and smoking, a significant correlation between non-alcoholic fatty liver disease (NAFLD) and atherosclerosis has been reported. Patients with NAFLD die from CVD more frequently than from liver disease, and NAFLD and its more severe condition, non-alcoholic steatohepatitis (NASH), are now considered independent, and likely causal, risk factors for the development of atherosclerosis. It has been therefore proposed that therapies that treat NAFLD may also inhibit atherogenesis, especially since these diseases share common underlying factors including inflammation, lipid deposition and oxidation, and dysfunctional macrophages. At MAX BioPharma we have identified a lead proprietary semi-synthetic oxysterol, Oxy210, that significantly inhibits NASH in a humanized mouse model, ApoE*3-Leiden.CETP, as evidenced by reduction in hepatic lipid deposition, inflammation, and fibrosis as well as reduced inflammatory cytokines in the circulation. Oxy210 is orally bioavailable and has favorable pharmacokinetic and safety profiles and is readily scalable. In preliminary studies we have found that Oxy210 has anti-inflammatory effects in macrophages treated with lipopolysaccharide or the atherogenic phospholipid, PGPC, through inhibition of toll-like receptor signaling. We hypothesize that given Oxy210’s inhibitory effects on NASH and inflammation, and given the common mediators of NASH and atherosclerosis, Oxy210 may also possess anti-atherosclerosis properties. In the present application we propose to test this hypothesis in vitro and in vivo through studies outlined in 3 specific Aims: 1) Examination of the effect of Oxy210 on the activation of HSCs, Kupffer cells and vascular endothelial cells treated with LPS or a synthetic TLR2 agonist in vitro; 2) Examination of the molecular mechanisms of action (MoAs) for anti-inflammatory effects of Oxy210 in vitro; and 3) Assessment of the effect of Oxy210 on atherogenesis in ApoE*3-Leiden.CETP transgenic humanized mouse model of NASH in vivo. Findings from the proposed studies will demonstrate the potential of Oxy210 as a first in class drug candidate for therapeutic development for the treatment of NASH and CVD.