CAT-5571 as a novel therapeutic that reduces infection and controls - PowerPoint PPT Presentation

CAT-5571 as a novel therapeutic that reduces infection and controls inflammation in cystic fibrosis Feng Liu 1 , Kathrin Krause 2 , Amal Amer 2 , Luanne Hall-Stoodley 2 , John F. Reilly 1 and Andrew J. Nichols 1 EPS1.05 1 Catabasis Pharmaceuticals, Inc. Cambridge, MA 02139, USA, 2 Department of Microbial Infection and Immunity, The Ohio State University, Columbus OH 43210, USA Autophagy • Depressed in cystic fibrosis • Critical component of immune regulation and host defense • Important for clearance of pathogens Beclin 1 LC3 *p<0.05 Human nasal mucosa from people with severe CF (n=10, homo or het Δ F508 CFTR) Luciani A, et al., Nat Cell Biol. (2010) 12: 863-75

CAT-5571 reduces pro-IL-1 b in P. CAT-5571 restores LC3 in macrophages CAT-5571 restores P. aeruginosa from Δ F508-CFTR mice to levels observed aeruginosa- infected mouse clearance in mouse Δ F508-CFTR in wild type mice Δ F508-CFTR macrophages macrophages *p<0.05 *p<0.05 *p<0.05 * Macrophage-associated CFU Pro-IL-1 b /GAPDH (A.U.) 5 3 * * (fold change from baseline) 4 2 3 2 1 1 0 0 Non-infected Vehicle CAT-5571 WT Vehicle CAT-5571 P. aeruginosa -infected CF By restoring autophagy, CAT-5571 CAT-5571 enhances bacterial clearance and blunts the hyperinflammatory addresses a fundamental defect in CF response in P. aeruginosa -infected macrophages that is present from birth Bacterial clearance and IL- 1β measurement in macrophages: WT and c ftr F508del/F508del mouse macrophages were treated for 24 hours with vehicle or 10 µM CAT-5571, then infected with P. aeruginosa PA01 with an MOI of 10:1 for 2 hours . Elimination of extracellular bacteria was performed by replacement with media containing 200 g/mL gentamicin. The cells were then incubated at 37 °C in 5% CO 2 until lysis at 4 hours post infection. CFU were determined by serial dilution and plating onto nutrient agar. For pro- iL1β is 6 hours post infection. Cells lysates were analyzed by immunoblotting with anti-pro- IL1β antibody. Values are mean ± SEM of four independent experiments. Statistical analyses were performed using two-way ANOVA

P. aeruginosa clearance by CAT-5571 CAT-5571 enhances clearance of was attenuated when autophagy CAT-5571 enhances clearance of B. cenocepacia in mouse Δ F508- M. abscessus in mouse Δ F508- was inhibited by beclin-1 knockdown CFTR macrophages CFTR macrophages in normal hBE cells *p<0.05 *p<0.05 vs. vehicle * Macrophage-associated CFU Macrophage-associated CFU 10 1.0 3 (fold change from baseline) (fold change from baseline) (fold change from baseline) * * Vehicle hBEC-associated CFU 8 0.8 CAT-5571 (10 m M) 2 6 0.6 4 * * 0.4 1 2 0.2 0 0 WT Vehicle CAT-5571 Vehicle CAT-5571 CAT-5571 0.0 non-target KD beclin-1 KD (1.25 µ M) (10 µ M) CF CAT- 5571’s effect on pathogen CAT-5571 enhances the clearance of multiple, difficult to treat clearance is mediated by beclin-1 pathogens affecting people with CF Beclin-1 Knock down experiment: Normal human primary hBE cells were transfected either with beclin-1 siRNA or non-targeting siRNA for 20 hours. At 24 hours prior to infection, cells were pre-treated with CAT-5571 (10 µM) and the vehicle. Cells were infected with P. aeruginosa Xen05 72 hours post transfection at MOI of 1:50 in media containing CAT-5571 and vehicle control. Elimination of extracellular bacteria was performed at 2 hours post infection by replacement with media containing 200 g/mL gentamicin. The epithelial cells were then incubated at 37 °C in 5% CO 2 until lysis at 4 hours post infection. CFU were determined by serial dilution and plating onto nutrient agar. hBE-associated CFU relative to invasion (fold change). Values are mean ± SEM (n =3). Statistical analyses were performed using one- way ANOVA followed by multiple comparison test (* p < 0.05)

CAT-5571: Breaking the Downward Spiral of CF Progression ‣ Novel Mechanism of Action ‣ Host-Directed Therapy – Activates depressed autophagy, restoring host – Potential to avoid typical bacterial resistance mechanisms defense while preventing hyper-inflammation ‣ Acts in Concert with other CF Therapies – Effective independent of CFTR mutation – Potential to augment efficacy of antibiotics ‣ Addresses Difficult to Treat Pathogens – Potential to work on top of CFTR correctors and potentiators – Pseudomonas ‣ Orally Administered – Burkholderia – Does not add to inhalational treatment burden – Non-tuberculous mycobacteria CAT-5571 Obstruction Obstruction Obstruction Respiratory Respiratory Respiratory Tract Tract Tract CFTR Infection Infection Infection Dysfunction Structural Pulmonary Bronchiectasis Respiratory damage Insufficiency Failure Inflammation Inflammation Inflammation CAT-5571