PBI-4050 Decreases Hepatic Stellate Cell Activation and Ameliorates
Fibrosis in Carbon Tetrachloride (CCl4)-Induced Hepatic Fibrosis Model
François Sarra-Bournet, Brigitte Grouix, Kathy Hince, Alexandra Felton, Mikaël Tremblay,
Shaun Abbott, Jean-Simon Duceppe, Boulos Zacharie, Pierre Laurin and Lyne Gagnon
PROMETIC BIOSCIENCES INC., LAVAL, QUÉBEC,CANADA
p=0.020
Untreated
TGF-β1
0.75
Statistics: Student’s t-test
p=0.004
0.75
0.50
0.25
CCl4
CCl4 +
CCl4 +
PBI-4050
PBI-4050
(100 mg/kg) (200 mg/kg)
1.00
p=0.036
NS
0.75
0.50
0.25
CCl4
CCl4 +
CCl4 +
PBI-4050
PBI-4050
(100 mg/kg) (200 mg/kg)
p=0.028
1.00
p=0.002
0.75
0.50
Control
CCl4 +
CCl4 +
PBI-4050
PBI-4050
(100 mg/kg) (200 mg/kg)
CCl4
p=0.0001
0.25
1.00
0.75
p=0.022
p=0.041
Sham
0.50
0.25
CCl4
CCl4 +
CCl4 +
PBI-4050
PBI-4050
(100 mg/kg) (200 mg/kg)
Control
CCl4
2.0
Relative Quantitation
0.25
0.00
TGF-β1
TGF-β1 + PBI-4050
CTGF
1.00
1.5
p=0.017
p=0.027
NS
1.0
0.5
0.0
Control
0.75
CCl4
CCl4+
CCl4+
PBI-4050
PBI-4050
(100 mg/kg) (200 mg/kg)
p=0.001
Magnification 100X
0.25
p=0.0001
Collagen Deposition
0.00
TGF-β1
TGF-β1 + PBI-4050
mRNA expression
1.25
PPARγ
p=0.015
1.5
1.0
0.5
NS
1.00
p=0.006
p=0.001
0.75
0.50
p=0.0001
0.25
Control
Untreated
TGF-β1
TGF-β1 + PBI-4050
1.5
p=0.0005
1.0
p=0.0001
0.5
0.0
0.00
0.0
Histomorphometry
2.0
% Collagen in Liver
Untreated
CCl4
CCl4 +
CCl4 +
PBI-4050
PBI-4050
(100 mg/kg) (200 mg/kg)
ug Hydroxyproline/mg Liver
0.50
CCl4 +
CCl4 +
PBI-4050
PBI-4050
(100 mg/kg) (200 mg/kg)
PPARγ
p=0.0041
1.25
CCl4
CCl4 + PBI-4050
p=0.006
Untreated
p=0.008
0.00
Control
0.50
Relative Quantitation
TIMP-1
1.25
0.00
0.00
Control
MMP-2
1.25
NS
Remodeling Markers
α-SMA Immunohistochemistry Staining
Relative Quantitation
Euthanasia and
fibrosis evaluation
(IHC, qPCR, histology)
p=0.008
p=0.023
0.00
Control
1.00
Relative Quantitation
Oral administration
of vehicle or PBI-4050
(100 or 200 mg/kg, day 1 to 58)
0.25
Snail1
1.25
2.0
Day 59 or 87
0.50
TGF-β1 + PBI-4050
p=0.012
Day 1
p=0.001
0.00
0.00
IP injection of 2 ml/kg of CCl4 10%
in olive oil, twice a week for 7 weeks
C57BL6/J male
0.75
p=0.004
1.00
CTGF (Day 87)
1.25
Relative Quantitation
p=0.028
1.00
p=0.005
Snail1
1.25
Relative Quantitation
0.50
Relative Quantitation
Relative Quantitation
0.75
0.25
α-SMA
1.25
1.00
Profibrotic/EMT Markers
Relative Quantitation
α-SMA
1.25
METHODS
In vitro human HSC activation: Serumstarved HSC cells were treated with or
without PBI-4050 (500 µM) and TGF-β1
(10 ng/ml) for 24 h, and qPCR analysis of
relative gene expression was performed
with TaqMan© Gene Expression Assay.
CCl4-induced liver fibrosis study:
PBI-4050 REDUCES FIBROSIS IN LIVER OF CCl4-TREATED MICE
PBI-4050 REDUCES TGF-β1INDUCED ACTIVATION OF HSC
Relative Quantitation
Hepatic stellate cells (HSC) are considered
a key player in the fibrogenic process of
the liver. In response to liver injury, HSC
become activated and transdifferentiate
into myofibroblast-like cells which display
increased proliferation and collagen
synthesis.
PBI-4050 is a first-in-class anti-fibrotic
compound presently in clinical phase II
trial in metabolic syndrome associated with
diabetes and in idiopathic pulmonary fibrosis.
In the present study, we investigated
whether treatment with PBI-4050 could
decrease in vitro HSC activation and prevent
CCl4-induced fibrosis in mice.
RESULTS
Relative Quantitation
BACKGROUND AND AIM
0.50
Hydroxyproline Content
0.45
NS
p=0.0381
0.40
0.35
p=0.0001
0.30
0.25
Control
CCl4
CCl4 +
CCl4 +
PBI-4050
PBI-4050
(100 mg/kg) (200 mg/kg)
Control
CCl4
CCl4 +
CCl4 +
PBI-4050
PBI-4050
(100 mg/kg) (200 mg/kg)
CONCLUSIONS
This study demonstrates that PBI-4050:
► Reduces HSC activation
► Reduces liver fibrosis by decreasing
overexpression/overproduction of:
 profibrotic/EMT markers
 remodeling markers
 collagen deposition
These results suggest
that PBI-4050 offers
the potential as
a novel therapy
for hepatic fibrosis.