Program Number TO021 – 50 th ERA-EDTA Congress – May 21, Oral treatment with a novel first-in-class anti-fibrotic compound PBI-4050 reduces hepatic steatosis and improves kidney function in the diabetic db/db mouse model. 1 Presented by: Dr. Lyne Gagnon AASLD Conference Washington November 2-4, 2013 Disclosure: ProMetic BioSciences Inc. Additional Information Also including the results on the effect of PBI-4050 in CCl 4 -induced liver fibrosis

2

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 PBI-4050 preclinical data in two liver fibrosis models 3 NASH-fibrosis model CCl 4 -induced steatohepatitis (Slides 17-27) Chronic model Uni-nephrectomized diabetic (db/db) mouse model (Slides 4-16)

Chronic model: Uni-nephrectomized diabetic (db/db) mouse model 4

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 PBI-4050 reduces serum glucose to the C57BL/6 and sham level 5 Serum glucose measured on 5-hour starved mice

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 Treatment with PBI-4050 increases glucose metabolism in oral glucose tolerance test at day Serum glucose measured on 16-hour starved mice

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 PBI-4050 reduces liver steatosis Liver Steatosis Db/dbDb/db + PBI-4050 p = 0.04

8 PBI-4050 reduces liver steatosis C57BL/6Db/dbDb/db + PBI

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 PBI-4050 reduces fibrotic markers expression in liver 9

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 Treatment with PBI-4050 reduces kidney hyperfiltration in db/db mice at day 97 10

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 Treatment with PBI-4050 reduces proteinuria 11

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 Treatment with PBI-4050 increases urinary creatinine excretion 12

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 PBI-4050 reduces kidney mesangium lesions 13

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 PBI-4050 reduces fibrotic markers expression in kidney 14

15 PBI-4050 reduces lipid peroxidation in kidney

Conclusions: PBI-4050 offers the potential as a novel therapy for hepatic steatosis in DKD 16 In liver  Reduces steatosis  Reduces fibrotic markers (TGF- , collagen 1, MMP-2 and TIMP-1 mRNA expression) In kidney  Reduces kidney hyperfiltration, proteinuria, albuminuria  Increases urinary creatinine excretion  Reduces histological lesions in the mesangium  Reduces fibrotic markers expression (IL-6, collagen 1, TIMP-1 and MMP-2 mRNA expression)  Reduces oxidative stress (lipid peroxidation) PBI-4050 has now enter into clinical program.

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, NASH-Fibrosis Model CCl 4 -induced steatohepatitis 17

CCl 4 -induced liver fibrosis: Effect of PBI-4050 Study design Day 1: Oral administration of vehicle or PBI-4050 (day 1 to day 59) IP injection of 2ml/kg of CCl4 10% in olive oil, twice a week for 8, 12 or 16 weeks Day 59: Euthanasia and analysis C57BL6/J male 18

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 Day 59: Evidence of pre-fibrosis  The effect of PBI-4050 was studied on the pre-fibrosis/ fibrosis development in CCl 4 -induced hepatic fibrosis.  Fibrosis was estimated with the measurement of hydroxyproline which is a direct measure of collagen (marker of fibrosis) in the liver (next slide). PBI-4050 reduces liver fibrosis. 19

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 PBI-4050 significantly reduces hepatic concentration of hydroxyproline 20 P = 0.007

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 PBI-4050 significantly reduces the histology activity index (HAI)-Knodell score  Intoxication with CCl 4 results in hepatocyte damage, necrosis, inflammation, and fibrosis, which spreads to link the vascular structures that feed into and drain the hepatic sinusoid (the portal tract and central vein radicle, respectively), and over 8-30 weeks results in the development of fibrosis to hepatocellular carcinoma.  Knodell score is the combined scores for necrosis, inflammation and fibrosis. Histology activity index (HAI) evaluates: 1)Periportal +/- bridging necrosis (bridging between portal-portal and portal-central linkage); 2)Intralobular degeneration (acidophil bodies, ballooning, focal necrosis- scattered foci of hepatocellular necrosis) and focal necrosis; 3)Portal inflammation, and 4)fibrosis (fibrous portal expansion, or bridging fibrosis (portal-portal or portal-central linkage or cirrhosis (loss of normal hepatic lobular architecture with fibrous septae separating and surrounding nodules)). 21

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 PBI-4050 significantly reduces the histology activity index (HAI)-Knodell score 22 p= 0.008

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 PBI-4050 significantly reduces the collagen score in liver (Masson’sTrichrome staining) 23 p = 0.03

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 PBI-4050 significantly reduces collagene deposition in CCl 4 -induced liver fibrosis (Masson’s trichrome stain)  The Masson’s trichrome stain the cytoplasm, keratin, muscle fibers and intracellular fibers in red colour; nuclei in black colour and collagen (fibrous tissue) in blue color.  All control mice revealed a normal distribution of collagen.  Extensive collagen deposition and bridging fibrosis were evident in CCl 4 -treated animals.  Significant reduction of collagen deposition and bridging formation between portal-portal or portal- central was observed in PBI-4050 treated animals. 24

25 Control CCL4) CCL4 CCL4 + PBI-4050 (200 mg/kg) PBI-4050 significantly reduces histological lesions in CCl 4 -induced liver fibrosis (Masson’s Trichrome)

Program Number TO021 – 50 th ERA-EDTA Congress – May 21, 2013 PBI-4050 significantly reduces histological lesions in CCl 4 -induced liver fibrosis (Hematoxylin-Eosin stain)  The hematoxylin eosin stains the cytoplasm in red colour; nuclei with blue colour and show the steatosis and lymphocyte infiltration in stained tissue.  A moderate steatosis and a severe inflammation are observed as compared to control (non-CCl 4 ) mice. Treatment with PBI-4050 reduces inflammatory infiltration in hepatic lobes. 26

27 Control CCL4) CCL4 CCL4 + PBI-4050 (200 mg/kg) PBI-4050 significantly reduces histological lesions in CCl 4 -induced liver fibrosis (Hematoxyline-Eosin)

28 Acknowledgements Biology Dr. Brigitte Grouix Lilianne Geerts François Sarra-Bournet Kathy Hince André Doucet Mikaël Tremblay Alexandra Felton Dr. Martin Leduc Liette Gervais Chemistry Dr. Christopher Penney Dr. Boulos Zacharie Dr. Shaun Abbott Jean-Simon Duceppe Frank Cesari Lyne Marcil Pierre Laurin