Liver Selective Acetyl-CoA Carboxylase Inhibition by ND-654 Improves Survival in Cirrhotic Rats with Hepatocellular Carcinoma Omeed Moaven 1 , Lan Wei 1 , Geraldine Harriman 2 , Jeremy Greenwood 3 , Sathesh Bhat 3 , William F. Westlin 2 , H. James Harwood 2 , Rosana Kapeller 2 , Danielle K. DePeralta 1 , Kenneth K. Tanabe 1 , Bryan C. Fuchs 1 1 Division of Surgical Oncology, Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, MA, 2 Nimbus Therapeutics, Cambridge, MA, 3 Schrodinger, New York, NY ND-654 Abstract #4452 PBS DEN ND-654 (10 mg/kg) ND-654 (30 mg/kg) ICAM-1/CD54 CD26/Dpp4 IL-1B IL-11 RANTES IL-6 IL-10 MCP-1/CCL2 Pref-1/DLK-1 RAGE TNF-α LIF TIMP-1 IGFBP1 IGFBP2 IGFBP3 IGFBP5 IGFBP6 FGF-21 0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 0 1 2 3 4 5 6 0 1 2 3 6 9 12 15 Serpin E1 Endocan Leptin Resistin 3 Fold Expression as Compared to PBS Control 0 0.00005 0.0001 0.0003 PBS DEN DEN + 10 ND-654 DEN+ 30 ND-654 mg/kg 0.0002 0.00025 0.00015 0 0.00005 0.0001 0.0003 PBS DEN DEN + 10 ND-654 DEN+ 30 ND-654 mg/kg 0.0002 0.00025 0.00015 0 0.0004 0.0005 0.0009 PBS DEN DEN + 10 ND-654 DEN+ 30 ND-654 mg/kg 0.0007 0.0008 0.0006 0.0001 0.0002 0.0003 * * * * * * Gene Expression (AU) Gene Expression (AU) Gene Expression (AU) OTHERS COO - Acetyl CoA Malonyl CoA BC CT Acetyl -CoA Acetyl -CoA Acetyl -CoA Malonyl -CoA Citrate Very long chain fatty acids ACC1 ACC2 Fatty Acids Fatty Acids FAO Pyruvate Oxaloacetate Krebs Cycle ETC NADH NADH + FADH 2 FAO MITOCHONDRION PEROXISOME Phosphoinositides Eicosanoids Sphingolipids Phosphoglycerides CELL GROWTH SIGNALING MEMBRANE BIOGENESIS AMPK LKB1 CPT1 0.000001 0.0001 0.01 1 0 5000 10000 15000 cpm ND-654 (μM) 0.01 0.1 1 10 100 Plasma Liver Muscle Tissue Concentration (μM) 0 20 40 60 80 0.03 3 DPMs/mg Tissue 1 0.3 0.1 Vehicle mg/kg 0 2 3 6 Vehicle 0.3 3 30 mg/kg 1 5 4 Malonyl-CoA (nmol/g tissue) 0 4 5 Vehicle 0.3 3 30 mg/kg 1 2 3 Malonyl-CoA (nmol/g tissue) ND-654 10 mg/kg qd po Control (PBS) 13 18 Sacrifice 50mg/kg DEN Weeks 0 8 Cirrhosis HCC 50mg/kg DEN X Fibrosis 0 100 200 300 400 500 600 Body Weight (g) PBS DEN DEN + ND-654 0 20 40 60 80 100 140 ALT (IU/L) PBS DEN DEN + ND-654 0 100 150 200 250 300 350 AST (IU/L) PBS DEN DEN + ND-654 120 50 # # # 0 0.2 0.3 0.4 0.5 0.6 0.7 Spleen Weight (% Body) PBS DEN DEN + ND-654 0 1 2 3 4 5 6 TBIL (mg/dL) PBS DEN DEN + ND-654 0 1.5 2 2.5 3 3.5 4.5 Alb (g/dL) PBS DEN DEN + ND-654 4 1 .5 0 20 40 60 80 100 160 TG (mg/dL) PBS DEN DEN + ND-654 120 # 0.1 140 # # ** ** 0 1 2 3 4 5 9 Liver Weight (% Body) PBS DEN DEN + ND-654 0 10 15 20 25 Tumor Nodules PBS DEN DEN + ND-654 5 # * 6 7 8 # ** PBS DEN DEN + ND-654 ND-654 10 mg/kg qd po 50mg/kg DEN 13 18 50mg/kg DEN Weeks 0 8 Cirrhosis HCC 50mg/kg DEN Fibrosis 0 10 20 30 40 50 60 70 80 90 100 90 95 100 105 110 115 120 125 130 135 140 145 150 155 Percent Survival Time (Days) Vehicle ND-654 10 ND-654 30 0 2 4 6 8 10 12 Vehicle PBS Collagen Area (%) DEN + 10 ND-654 DEN+ 30 ND-654 mg/kg Vehicle qd po ND-654 30 mg/kg qd po Vehicle/ DEN ND-654 10 mg/kg ND-654 30 mg/kg MEDIAN SURVIVAL 114 days 126 days 129 days P VALUE 0.02 0.009 PCNA Actin DEN DEN + ND-654 Tumor PBS DEN DEN + ND-654 Liver PCNA Actin DEN DEN + ND654 H&E PCNA Cleaved Caspase-3 2. Nimbus Solves ACC Druggability Challenge by Targeting Allosteric Site in Biotin Carboxylase Domain 1. ACC: Emerging Tumor Metabolism Target: Tumors Up-regulate ACC to Fuel Cell Growth 3. ND-654: Hepato-Selective ACC Inhibitor 4. ND-654 Demonstrates Liver Tissue Specifc Target Engagement and Pharmacology 5. ND-654 is Well Tolerated in Rats Administered DEN Animals on Drug Appear Healthy 6. Liver Directed ND-654 Improves Markers of Liver Health in DEN Treated Rats 7. Treatment with 10 mpk ND-654 QD Results in Marked Reduction of Tumor Nodules 8. ND-654 Decreases PCNA Expression in Tumors and Results in Tumor Necrosis 11. Quantitative Liver Adipokine Array From ND-654 Treated DEN Rats Demonstrates Modulation of Fibrosis, Infammation and Tumor Markers in the Liver 10. ND-654 Shows Improvements in Fibrosis, Stellate Cell Activation and Infammation Markers in the Rat DEN Model 9. ND-654 Signifcantly Improved Survival in the Aggressive DEN HCC Model OVERVIEW CONCLUSION 18 Week DEN Rat Study 18 Week DEN Rat Study 18 Week DEN Rat Study 18 Week DEN Rat Study DEN Rat Survival Study DEN Rat Survival Study DEN Rat Survival Study • Current treatment options for HCC are limited, and as such, prognosis is extremely poor with a 5-year survival less than 12% • Metabolic attenuation is a promising approach to cancer therapy, and rate-limiting steps in key biosynthetic pathways are particularly attractive targets. HepaSpecifc ND-654 is a potent allosteric inhibitor of Acetyl-CoA Carboxylase (ACC) • ND-654 demonstrates excellent PK/PD relationships in the target tissue, the liver (ex- posure vs Malonyl-CoA reduction) and is effective at modifying Cirrhotic and HCC relevant endpoints in the DEN model. Herein, we report on two rat DEN studies. An 18 week fxed end point study utilizing 10 mg/kg ND-654 PO, QD and a sur- vival study utilizing 10 & 30 mg/kg ND-654 PO, QD. • ND-654 demonstrates the ability for tissue targeted ACC inhibition to decrease tu- mor burden, liver fbrosis and prolong survival in the rat DEN model of HCC • ND-654 preferentially biodistributes to the liver where it inhibited ACC activity but did not increase liver toxicity. • ND-654 inhibits HCC development by decreasing tumor proliferation and causing tu- mor necrosis. • ND-654 reduces markers of infammation and fbrosis and improves overall survival in the DEN rat model. • Our results therefore provide further evidence that de novo lipogenesis is an import- ant mediator of hepatic carcinogenesis. • Selective inhibition of hepatic ACC is a potential therapeutic strategy for HCC Infammation Fibrosis Tumor & Lipids DEN DEN +ND-654 10 PBS Alpha-Smooth Muscle Actin CD68 Infammation Marker Collagen 1A1 Marker (Fibrosis) DEN +ND-654 30 α-SMA Trichrome • Only potent BC domain inhibitor: Soraphen A • Industry efforts on analoging soraphen proved unsuccessful • Allosteric binding site implicated in dimerization of ACC • Highly lipophilic binding site • Inhibitors bind to active site • Past programs exhibited poor drug-like properties 1. Novel class of ACC inhibitors 2. Prototype for modulating protein-protein interactions Nimbus inhibitors prevent ACC dimerization and activation ND-630 binds to the same site as Soraphen A Nimbus Breakthrough Challenges in Drugging ACC Target potency • ACC1 IC 50 = 3 nM • ACC2 IC 50 = 8 nM • HepG2 EC 50 = 4 nM (serum free) = 14 nM (10% serum) Pharmacology • Rat FASyn ED 50 = 0.3 mg/kg PO • Rat Malonyl-CoA ED 50 = 0.3 mg/kg (liver) • Rat DEN HCC MED = 10 mg/kg ADME • Moderate multispecies intrinsic clearance (human, mouse, rat, dog, monkey) • P450 inhib >50 μM • Highly selective across broad panel (>1000x) Exposure at 10 mg/kg Hep-G2 Cell FASyn Inhibition [ 14 C]acetate incorporation into FA • NSCLC: Targeted opportunity via LKB1- AMPK-ACC axis - LKB1 inactivated in Peutz–Jeghers syndrome, NSCLC and cervical carcinoma • Breast Cancer: ACC1 knock down causes apoptosis in tumor cells - BRCA1 stabilizes inactive phosphorylated form of ACC - BRCA1 also implicated in prostate cancer • RCC: ACC overexpression - Associated with worse survival in clear cell Renal Cell Carcinoma • HCC: ACC overexpression - ACC expression and de novo lipogenesis are in- creased in HCC ACC Oncology Validation Liver p < 0.01 Muscle (Gastrocnemius) n.s. • ND-654 was administered orally to DEN treated rats once a day (6/7 days per week) • Drug is well tolerated, no change in body weights, ALT and AST • ND-654 decreased serum triglycerides and total bilirubin 25 First St #404, Cambridge, MA 02141, USA • www.nimbustx.com • Tel: 857.999.2009 • info@nimbustx.com