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

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)

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

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

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

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)

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

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

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

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)

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)

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)

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)

Many thanks to all involved Galapagos colleagues