1. Strong reversal of the lung fibrosis disease signature by autotaxin inhibitor GLPG1690 in a mouse model for IPF
Maté Ongenaert, PhD
Senior Scientist Bioinformatics
Sonia Dupont, Roland Blanqué, Reginald Brys, Ellen van der Aar, Bertrand Heckmann
ERS London, UK September 6th 2016

2. Disclosure
I and other authors have the following real or percieved conflicts of interest that relate to this presentation:
All employees of Galapagos NV (Mechelen, Belgium) or Galapagos SASU (Romainville, France)

3. Outline Autotaxin/LPA pathway background
Mouse bleomycin (BLM) model - GLPG1690 performance and target engagement
Transcriptomics studies in BLM model
Summary and outlook N

4. Autotaxin (ATX) Target background
GT2645
ATX
Lysophosphatidylcholine (LPC)
Lysophosphatidic acid (LPA)
Also known as ENPP2, secreted enzyme
Widely expressed (highest in brain, lymph nodes, kidney and testis)
Converts LPC to the bioactive lipid mediator LPA
“LPA” covers a family of related molecules (i.e. LPA18:2, LPA20:4)
ATX is main source of LPA in blood

5. LPA signalling
LPA acts through at least six distinct G-protein-coupled receptors (LPA1–6)
LPA controls activities like migration, contraction & survival
Studies with KO of LPA receptors indicate a role in
bone development
fertility/reproduction
neurogenesis
formation of blood- and lymphatic vessels
Stoddard and Chun, 2015

6. ATX/LPA pathway and IPF Preclinical and translational validation
In modeled disease: bleomycin-exposed mice
increased LPA in BALF (Tager et al, 2008)
ATX levels increased in lungs (Oikonomou, 2012)
inhibition of ATX attenuated lung fibrosis in mice (Oikonomou, 2012)
genetic or pharmacological inhibition of LPAR1 attenuates development of pulmonary fibrosis (Tager et al, 2008; Swaney et al, 2010)
In patients with idiopathic pulmonary fibrosis (IPF)
LPA levels increased in BALF (Tager et al, 2008)
ATX levels elevated in lung (Oikonomou et al, 2012)
LPA increased in exhaled breath condensate (Montesi et al, 2014)

7. GLPG1690 in vivo activity
Mouse bleomycin model for lung fibrosis (prophylactic)
Relative levels of LPA species in BALF of mice subjected to BLM treatment
LPA 17:0 peak area
LPA peak area /
Ashcroft score
Group
vehicle
BLM + vehicle
BLM + pirfenidone 50 mg/kg bid
BLM + ‘1690  30 mg/kg bid
GLPG1690 reduces fibrosis in BLM models, reduces LPA levels in BALF

8. GLPG1690 - bleomycin model Global transcriptomics (PCA)
vehicle
bleomycin
GLPG1690
Lung microarray profiling: GLPG1690 protects for BLM induced effects on a global transcriptome level

9. GLPG1690 - bleomycin model Most affected genes
vehicle
BLM
‘1690
Top-40 most differentially expressed genes
perfect separation of the BLM cluster from sham
GLPG1690 partially reverses the BLM effect

10. GLPG1690 - bleomycin model Relevance in model
Strong negative correlation (Spearman R=-0.74) between BLM effect and GLPG1690 effects
BLM
(UP)
‘1690
(DOWN)
1088
probes
259 18
^ Probes with both:
|log2(FC)|>1
FDR< 1%
BLM (log2 Fold change)
‘1690 (log2 Fold Change)

11. GLPG1690 - bleomycin model Functions
immune response
cytokines, Jak-Stat, TGFb
Affected functions and pathways by BLM and counteracted by GLPG1690 relevant in BLM model and IPF biology:
extracellular matrix (Tnc, Spp1)
several collagens (Col3a1, Col5a1)
cytokines / chemokines (Cxcl12, Ccl2)
Cxcl12
Col1a2
Serpine2
Smad6
Smad6
Extracellular matrix – focal adhesion
Col1a2
Ednrb
Col5a1
Tnc
Tnc
Ccl2
Serpine2
Color: log2 FC
Area: log2 FC * -log10(p-value)

12. GLPG1690 - bleomycin model BLM/GLPG1690 – IPF relevance
Gene Symbols
BLM mouse models
GLPG1690
Human IPF (public data)
Mouse
Human
BLM this study
BLM GSE40151
GLPG1690 this study
GSE32537
GSE10667
GSE53845
Yang et al.
Cxcl12
CXCL12
3.09
1.11
-1.4
1.17
2.21
1.96
1.5
Col3a1
COL3A1
2.51
1.15
-0.71
1.4
2.82
1.71
2.19
Spp1
SPP1
4.19
1.59
-1.56
1.61
3.67
3.49
1.77
Tnc
TNC
4.6
2.54
-2.22
1.13
0.86
1.89
1.49
52 genes upregulated in BLM and at least in ¾ IPF vs. normal studies and strongly counteracted by ‘1690
BLM: log2(FC)>1
counteracted by ‘1690: log2(FC)< -1
IPF studies: log2(FC)>0.5 in at least ¾ studies
Many of the genes identified are relevant in IPF and/or altered in expression in IPF patients (assessed in 4 public transcriptomics studies)

13. Summary and outlook
GLPG1690: potent and selective inhibitor of autotaxin
Strong anti-fibrotic effects in the mouse bleomycin model
IPF-related gene expression signature clearly affected after GLPG1690 administration
Pharmacological and literature data support positioning in IPF
Exploratory phase IIa study in IPF patients ongoing (FLORA)

14. Many thanks to all involved Galapagos colleagues