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

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.