Mitochondrial Uncoupler HU 6 Reduces Hepatic Oxidative Stress

Mitochondrial Uncoupler HU 6 Reduces Hepatic Oxidative Stress in the DIAMOND™ Mouse Model Thus Abrogating NASH Development. Francisco Portell 3, Stephen Eaton 3, Jameel Dennis 3, Rebecca Caffrey 1, Jonathon Marioneaux 1, Isaac Onyango 3, Omer Khan 3, Shaharyar Khan 3 and Arun Sanyal 2 1. Sanyal biotechnology LLC, Norfolk, VA, United States. 2. Virginia Commonwealth University, Richmond, VA, United States. 3. Gencia INTRODUCTION RESULTS Vehicle Control H&E Mouse ID 495 – 10 X The Gencia mitochondrial uncoupler HU 6 was evaluated for utility in treating NASH in a dietinduced animal model of metabolic syndrome and NASH (DIAMOND™ mice); development of NASH in this model is driven by insulin resistance, oxidative stress, inflammation, and leaky gut induced by Western Diet (WDSW). . AIM The aim of this study was to investigate effects of HU 6 on ROS formation and correlate these with disease progression. METHOD • Mice were grouped into 5 groups: vehicle control (VC), high dose HU 6 (HD), 5 mg/kg), low dose HU 6 (LD, 1 mg/kg), WDSW positive (PC) and NCNW negative (NC) natural history controls. • Mice were raised for 12 weeks on diet, corresponding to a baseline NASH F 0 in the WDSW groups. Treatment groups were then gavaged once daily with aqueous vehicle or drug in vehicle for 8 weeks. • Serum and liver tissue were harvested, snap frozen and fixed, and LFTs, blood lipids and histology on FFPE section stained with H&E and Sirius Red was performed. • DNA/RNA oxidative damage was measured via ELISA from representative liver samples. Key chemokines were also assayed from mouse livers utilizing Luminex Bead-based multiplex assays. Serum ALT, AST and ALP levels in the HD group were significantly lower than in the VC (P=0. 001, 0. 0004, and 0. 0002, respectively) and PC (P=0. 03, 0. 01, and 0. 00008, respectively) groups. High Dose H&E Mouse ID 619 – 10 X In the HD group the significant reduction in ballooning (P=0. 0000001), lobular inflammation and liver transaminotransferase levels is accompanied by a 72% reduction in MCP-1, 37% reduction in IP-10, 50% reduction in MCP-3 and a 43% reduction in MIP-2. Additionally, HU 6 -treated mice have a near 25% reduction in baseline oxidative damage to nucleic acids. Steatosis percentage and grade (P=0. 001 and 0. 01, respectively) were reduced in the HD group, which correlated with significantly lower body weights and liver weights compared to the VC and PC groups. Composite histology scores (NAS and SAF activity scores) were also significantly improved in the HD group compared to VC and PC groups. While all VC and PC mice progressed to NASH, HU 6 at 5 mg/kg prevented NASH development in all but one mouse. Blood chemistry Histology Scores Vehicle Control Sirius Red Mouse ID 495 – 10 X High Dose Sirius Red Mouse ID 619 – 10 X Low Dose H&E Mouse ID 619 – 10 X Low Dose Sirius Red Mouse ID 619 – 10 X Positive Control H&E Mouse ID 574 – 10 X Positive Control Sirius Red Mouse ID 574 – 10 X Negative Control H&E Mouse ID 699 – 10 X Negative Control Sirius Red Mouse ID 699 – 10 DNA/RNA Oxidative damage Chemokine/Cytokine IP-10 MIP-2 CONCLUSIONS With only eight week of dosing low levels of HU 6, measures of oxidative stress and chemokines, correlating with decreases in ballooning, inflammation, and progression from simple steatosis to NASH, were significantly reduced. HU 6 also produced significant reductions in body and liver weight, correlating with decreased hepatic steatosis. HU 6 impacted multiple drivers of NASH in this study, thus supporting the rationale for further study of HU 6’s therapeutic potential in NASH. MCP-1 MCP-3 CONTACT INFORMATION Francisco Portell frportell@genciabiotech. com Rebecca Caffrey rebecca@sanyalbio. com REFERENCES • Mansouri A, Gattolliat CH, Asselah T. Mitochondrial Dysfunction and Signaling in Chronic Liver Diseases. Gastroenterology Vol. 155, pp 629 -647. 2018