Transcript Oral treatment with PBI-4050, a novel first-in-class anti

Additional Information
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.
Presented by: Dr. Lyne Gagnon
AASLD Conference
Washington
November 2-4, 2013
Also including the results on the effect of
PBI-4050 in CCl4-induced liver fibrosis
Disclosure: ProMetic BioSciences Inc.
Program Number TO021 – 50th ERA-EDTA Congress – May 21, 2013
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PBI-4050 preclinical data in two liver fibrosis
models
Chronic model
Uni-nephrectomized diabetic (db/db) mouse model
(Slides 4-16)
NASH-fibrosis model
CCl4-induced steatohepatitis
(Slides 17-27)
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Chronic model: Uni-nephrectomized diabetic
(db/db) mouse model
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PBI-4050 reduces serum glucose to the
C57BL/6 and sham level
Serum glucose measured on 5-hour starved mice
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Treatment with PBI-4050 increases glucose
metabolism in oral glucose tolerance test at day 112
Serum glucose measured on 16-hour starved mice
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PBI-4050 reduces liver steatosis
Liver Steatosis
3.0
2.5
p = 0.04
2.0
1.5
1.0
0.5
0
Db/db
Db/db + PBI-4050
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PBI-4050 reduces liver steatosis
C57BL/6
Db/db
Db/db + PBI-4050
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PBI-4050 reduces fibrotic markers expression
in liver
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Treatment with PBI-4050 reduces kidney
hyperfiltration in db/db mice at day 97
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Treatment with PBI-4050 reduces proteinuria
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Treatment with PBI-4050 increases urinary
creatinine excretion
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PBI-4050 reduces kidney mesangium lesions
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PBI-4050 reduces fibrotic markers expression
in kidney
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PBI-4050 reduces lipid peroxidation in kidney
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Conclusions: PBI-4050 offers the potential as a
novel therapy for hepatic steatosis in DKD
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.
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NASH-Fibrosis Model
CCl4-induced steatohepatitis
Program Number TO021 – 50th ERA-EDTA Congress – May 21, 2013
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CCl4-induced liver fibrosis: Effect of PBI-4050
Study design
IP injection of 2ml/kg of CCl4 10% in olive
oil, twice a week for 8, 12 or 16 weeks
C57BL6/J male
Day 1: Oral administration of vehicle or PBI-4050
(day 1 to day 59)
Day 59: Euthanasia
and analysis
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Day 59: Evidence of pre-fibrosis
 The effect of PBI-4050 was studied on the pre-fibrosis/
fibrosis development in CCl4-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.
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PBI-4050 significantly reduces hepatic
concentration of hydroxyproline
P = 0.007
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PBI-4050 significantly reduces the histology
activity index (HAI)-Knodell score
 Intoxication with CCl4 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 necrosisscattered 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)).
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PBI-4050 significantly reduces the histology
activity index (HAI)-Knodell score
p= 0.008
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PBI-4050 significantly reduces the collagen score
in liver (Masson’sTrichrome staining)
p = 0.03
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PBI-4050 significantly reduces collagene deposition in
CCl4-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 CCl4-treated animals.
 Significant reduction of collagen deposition and
bridging formation between portal-portal or portalcentral was observed in PBI-4050 treated animals.
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PBI-4050 significantly reduces histological lesions in
CCl4-induced liver fibrosis (Masson’s Trichrome)
Control
CCL4)
CCL4 + PBI-4050 (200 mg/kg)
Control
CCL4
CCL4 + PBI-4050 (200 mg/kg)
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PBI-4050 significantly reduces histological lesions in
CCl4-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-CCl4) mice.
Treatment with PBI-4050 reduces inflammatory
infiltration in hepatic lobes.
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PBI-4050 significantly reduces histological lesions in
CCl4-induced liver fibrosis (Hematoxyline-Eosin)
Control
CCL4)
CCL4 + PBI-4050 (200 mg/kg)
Control
CCL4
CCL4 + PBI-4050 (200 mg/kg)
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Acknowledgements
Biology
Dr. Brigitte Grouix
Lilianne Geerts
François Sarra-Bournet
Kathy Hince
André Doucet
Mikaël Tremblay
Alexandra Felton
Dr. Martin Leduc
Liette Gervais
Frank Cesari
Lyne Marcil
Pierre Laurin
Chemistry
Dr. Christopher Penney
Dr. Boulos Zacharie
Dr. Shaun Abbott
Jean-Simon Duceppe
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