Liên Lepescheux

Preclinical Characterization of GLPG0634, a Selective Inhibitor of JAK1, for the Treatment of Inflammatory Diseases

The JAKs receive continued interest as therapeutic targets for autoimmune, inflammatory, and oncological diseases. JAKs play critical roles in the development and biology of the hematopoietic system, as evidenced by mouse and human genetics. JAK1 is critical for the signal transduction of many type I and type II inflammatory cytokine receptors. In a search for JAK small molecule inhibitors, GLPG0634 was identified as a lead compound belonging to a novel class of JAK inhibitors. It displayed a JAK1/JAK2 inhibitor profile in biochemical assays, but subsequent studies in cellular and whole blood assays revealed a selectivity of ∼30-fold for JAK1- over JAK2-dependent signaling. GLPG0634 dose-dependently inhibited Th1 and Th2 differentiation and to a lesser extent the differentiation of Th17 cells in vitro. GLPG0634 was well exposed in rodents upon oral dosing, and exposure levels correlated with repression of Mx2 expression in leukocytes. Oral dosing of GLPG0634 in a therapeutic set-up in a collagen-induced arthritis model in rodents resulted in a significant dose-dependent reduction of the disease progression. Paw swelling, bone and cartilage degradation, and levels of inflammatory cytokines were reduced by GLPG0634 treatment. Efficacy of GLPG0634 in the collagen-induced arthritis models was comparable to the results obtained with etanercept. In conclusion, the JAK1 selective inhibitor GLPG0634 is a promising novel therapeutic with potential for oral treatment of rheumatoid arthritis and possibly other immune-inflammatory diseases.

Triazolopyridines as Selective JAK1 Inhibitors: From Hit Identification to GLPG0634

Janus kinases (JAK1, JAK2, JAK3, and TYK2) are involved in the signaling of multiple cytokines important in cellular function. Blockade of the JAK-STAT pathway with a small molecule has been shown to provide therapeutic immunomodulation. Having identified JAK1 as a possible new target for arthritis at Galapagos, the compound library was screened against JAK1, resulting in the identification of a triazolopyridine-based series of inhibitors represented by 3. Optimization within this chemical series led to identification of GLPG0634 (65, filgotinib), a selective JAK1 inhibitor currently in phase 2B development for RA and phase 2A development for Crohns disease (CD).

Characterization of GLPG0492, a selective androgen receptor modulator, in a mouse model of hindlimb immobilization

BackgroundMuscle wasting is a hallmark of many chronic conditions but also of aging and results in a progressive functional decline leading ultimately to disability. Androgens, such as testosterone were proposed as therapy to counteract muscle atrophy. However, this treatment is associated with potential cardiovascular and prostate cancer risks and therefore not acceptable for long-term treatment. Selective Androgen receptor modulators (SARM) are androgen receptor ligands that induce muscle anabolism while having reduced effects in reproductive tissues. Therefore, they represent an alternative to testosterone therapy. Our objective was to demonstrate the activity of SARM molecule (GLPG0492) on a immobilization muscle atrophy mouse model as compared to testosterone propionate (TP) and to identify putative biomarkers in the plasma compartment that might be related to muscle function and potentially translated into the clinical space.MethodsGLPG0492, a non-steroidal SARM, was evaluated and compared to TP in a mouse model of hindlimb immobilization.ResultsGLPG0492 treatment partially prevents immobilization-induced muscle atrophy with a trend to promote muscle fiber hypertrophy in a dose-dependent manner. Interestingly, GLPG0492 was found as efficacious as TP at reducing muscle loss while sparing reproductive tissues. Furthermore, gene expression studies performed on tibialis samples revealed that both GLPG0492 and TP were slowing down muscle loss by negatively interfering with major signaling pathways controlling muscle mass homeostasis. Finally, metabolomic profiling experiments using 1H-NMR led to the identification of a plasma GLPG0492 signature linked to the modulation of cellular bioenergetic processes.ConclusionsTaken together, these results unveil the potential of GLPG0492, a non-steroidal SARM, as treatment for, at least, musculo-skeletal atrophy consecutive to coma, paralysis, or limb immobilization.

THU0116 Biological Effects of the JAK1 Selective Inhibitor GLPG0634 on Inflammation Markers in Arthritic Mice

Background Non-selective Janus kinase (JAK) inhibitors have shown long-term efficacy in treating rheumatoid arthritis (RA). However, tolerated doses and thereby efficacy are limited due to JAK2-driven side effects. Selective inhibition of JAK1 may be associated with an improved safety profile while maintaining clinical efficacy. Currently available information for JAK1 inhibitors is limited. GLPG0634 is a JAK inhibitor which displays a high selectivity for JAK1 over JAK2 in human whole blood assays. It is currently being developed for RA treatment and displayed good efficacy and tolerability in a 4-week Phase 2a study. Objectives Characterize the mode of action of a JAK1-selective inhibitor by measuring the impact of GLPG0634 on JAK pathway activity and inflammation parameters in blood and paws in the mouse CIA model. Methods Mice with established collagen-induced arthritis (CIA) were treated with GLPG0634 (50 mg/kg po, bid) or etanercept (10 mg/kg ip, 3 times a week). Disease progression was evaluated using clinical score and histological parameters. Expression of genes related to JAK pathways and markers of inflammation were measured in hind paws as well as in circulating leukocytes (WBC) using qRT-PCR. Luminex technology was used to measure a panel of 35 cytokines and chemokines in mouse serum. Results Oral treatment with GLPG0634 decreased inflammation in CIA in mice as measured by clinical scores and paw histology, with the same efficacy as etanercept. In paws of arthritic mice, an increase of 20- to more than 200-fold in the expression of inflammation markers (IL-6, IL-1β, TNFα, CXCL1) was found. A 30% to 45% decrease of their expression was observed in GLPG0634-treated arthritic mice. A similar expression pattern was observed for the osteoclast differentiation factor RANKL and for proteases linked to inflammation (MMP3, MMP13). These findings corroborate the bone and cartilage protective effects of GLPG0634 observed by histological analysis of paws. In addition, decreases from 30 to 70% in serum levels of cytokines and chemokines (IL-6, IP-10/CXCL10, MCP-1) in GLPG0634-treated arthritic mice confirmed the immunomodulation and anti-inflammatory effects. Of interest, the increased expression of JAK1-induced genes Mx1 and Mx2 in arthritic mice both in WBC and hind paws was abolished after treatment of animals with GLPG0634, whereas expression of the JAK2-dependent gene HRH4 was not altered by GLPG0634 treatment. These findings show that the decrease in inflammation markers was associated with inhibition of JAK1 but not JAK2. Conclusions These data show that oral GLPG0634 administration reduces inflammation in the mouse CIA model to the same extent as parenteral etanercept. Effects on several inflammation markers in paws and serum demonstrate the anti-inflammatory activity of GLPG0634. These effects were achieved through selective JAK1 inhibition. These data establish that selective JAK1 inhibition by GLPG0634 is sufficient to mediate strong efficacy in an established arthritis model and provide some useful potential markers to follow pathology progression in arthritic mice and humans. Disclosure of Interest B. Vayssiere Employee of: Galapagos SASU, S. de Vos Employee of: Galapagos NV, D. Merciris Employee of: Galapagos SASU, M. Auberval Employee of: Galapagos SASU, S. Dupont Employee of: Galapagos SASU, N. Vandeghinste Employee of: Galapagos NV, L. Lepescheux Employee of: Galapagos SASU, P. Clement-Lacroix Employee of: Galapagos SASU, P. Delerive Employee of: Galapagos SASU, L. van Rompaey Employee of: Galapagos NV, R. Brys Employee of: Galapagos NV, R. Galien Employee of: Galapagos SASU

OP0180 Pharmacological characterization of the ADAMTS-5 inhibitor GLPG1972: an oral anti-catabolic agent for the treatment of osteoarthritis

Background Degradation of articular cartilage and alterations of the underlying subchondral bone are hallmarks of osteoarthritis (OA)1. A disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMTS-5) is a key aggrecan-cleaving enzyme involved in this pathogenic process from the earliest stages of cartilage degradation2 and as such, is an attractive drug target for the development of a disease-modifying OA drug (DMOAD)3. Objectives In this report we describe the in vitro and in vivo characterization of the small molecule GLPG1972, an inhibitor of ADAMTS-5. GLPG1972 anti-catabolic activity was evaluated in murine and human cartilage explants and DMOAD activity was investigated in the destabilization of the medial meniscus (DMM) mouse model4. Methods The ADAMTS-5 biochemical assay is based on the cleavage of a fluorescent substrate by recombinant ADAMTS-5. Mouse femoral head cartilage explants were stimulated by interleukin-1a (IL-1a) for 3 days and GAG release quantified2b. Human articular cartilage explants were stimulated with IL-1β for 12 or 19 days and the NITEGE epitope quantified using the AGNx1 assay. Unilateral OA was induced in C57BL6 mice by DMM4. Mice were treated with vehicle or GLPG1972 at 30, 60 or 120 mg/kg, b.i.d. for 8 weeks. Medial femorotibial joint sections were scored by an evaluator blinded to treatment. Results GLPG1972 showed potent inhibition of human ADAMTS-5 (IC50=20 nM). Inhibition of ADAMTS-4 was moderate (IC50=57 nM), and selectivity over 100-fold was observed against a large panel of zinc metalloproteinases. GLPG1972 displayed potent anti-catabolic activity in cartilage explants, with IC50 values being 2 μM and <1 μM in mouse and human, respectively. In the DMM mouse model, GLPG1972 demonstrated DMOAD activity, as shown by significant reduction of femorotibial cartilage proteoglycan loss and cartilage damage score, as well as significant impact on subchondral bone sclerosis. Conclusions GLPG1972 is an orally bioavailable, potent and selective ADAMTS-5 inhibitor showing significant anti-catabolic activity in cartilage explants. In the DMM model, treatment with GLPG1972 resulted in significant protective effects on both cartilage and subchondral bone pathology. Taken together these results provide support to progress GLPG1972 into the clinic as an oral treatment for OA. References Hunter, D.J., et al. Curr. Opin. Rheumatol. 2009, 21,110–117. a) Glasson, S.S., et al Nature. 2005, Vol 434: 644–8; b) Stanton, H., et al. Nature. 2005, Vol 434: 648–5; c) Little, C.B., et al. Journal of Clinical Investigation. 2007, Vol 117(6):1627–36. Larkin, J. et al. Osteoarthritis Cartilage. 2015, 23 (8): 1254–1266. Little, C.B. and Hunter, D.J. Nat. Rev. Rheumatol. 2013, 9(8):485–497. Disclosure of Interest None declared

THU0138 Glpg0634m1, A Major Metabolite of the Jak1-Selective Inhibitor Glpg0634, is Also Jak1-Selective and Efficient in the Rat CIA Model

Background GLPG0634 is a JAK inhibitor displaying a high selectivity for JAK1 over JAK2 in human whole blood assays. It is currently being developed as an oral treatment for rheumatoid arthritis (RA) and showed good efficacy and tolerability in two 4-week Phase 2a RA studies. GLPG0634 has a major metabolite, GLPG0634m1, with a half-life which might contribute to its clinical efficacy in RA patients. Objectives Characterize the pharmacological properties of the GLPG0634m1, its activity on JAK-driven pathways and its efficacy in the rat CIA (collagen-induced arthritis) model. Methods Selectivity of the metabolite GLPG0634m1 was measured in vitro in radiometric recombinant kinase assays and ex vivo in whole blood assays (human, dog and monkey) by monitoring STAT phosphorylation by flow cytometry. Rats with established arthritis were treated with the GLPG0634m1 molecule (60 mg/kg po, QD) or etanercept (10 mg/kg ip, 3 times a week). The clinical score as well as histological parameters were used to monitor/quantify disease progression. Plasma concentration of the molecule was quantified by LC-MS/MS. Results Biochemical analysis of potency of GLPG0634m1 on recombinant JAK kinases showed that this compound is 10-fold less active against JAK1 and JAK2 than GLPG0634 with IC50s of 546 nM and 624 nM, respectively. Potency on JAK3 and TYK2 over 3 μM indicates that this molecule is more selective for JAK1 and JAK2 compared to JAK3 and TYK2. In human whole blood assays (WBA), GLPG0634m1 inhibited a JAK1-dependent event (IL-6-induced STAT1 phosphorylation) with an IC50 of 11.9 μM and a JAK2-dependent event (GMCSF-induced STAT5 phosphorylation) with an IC50 exceeding 100 μM, revealing the >10-fold selectivity of this molecule for JAK1 over JAK2. The JAK1 potency was confirmed with assays using IL2-induced STAT5 and IFNα-induced STAT1 phosphorylation, respectively triggering the JAK1/JAK3 and JAK1/TYK2 pathways. When given orally to rats with established arthritis, GLPG0634m1 displayed a pronounced efficacy comparable to etanercept, strongly reducing the impact of disease on e.g. paw swelling and Larsen score. These effects were confirmed at histological level with decreased pannus severity, bone and cartilage lesion as well as cell infiltration indexes. Plasma levels of GLPG0634m1 exceeded its WBA-derived JAK1 IC50 but were far below its WBA-derived JAK2 IC50, implying that the metabolite efficacy in the CIA model is driven by JAK1 inhibition, as observed previously with the parent molecule. Conclusions While displaying a lower potency compared to its parent molecule, GLPG0634m1 has a similar JAK1 selectivity. Oral administration of this metabolite, albeit at a 10-times higher dose than GLPG0634, reduces inflammation in the rat CIA model to the same extent as parenteral etanercept, an effect that is supported by JAK1 inhibition but appears independent of JAK2 inhibition. These findings, together with the high exposure and long half-life of this metabolite observed in phase 1 and phase 2 clinical studies in humans, strongly suggest that it may contribute to the clinical efficacy of the parent compound GLPG0634 in RA. Disclosure of Interest : C. Belleville-Da-Costa Grant/research support: Abbvie, Employee of: Galapagos SASU, D. Merciris Grant/research support: Abbvie, Employee of: Galapagos SASU, B. Vayssière Grant/research support: Abbvie, Employee of: Galapagos SASU, N. Houvenaghel Grant/research support: Abbvie, Employee of: Galapagos NV, A. Monjardet Grant/research support: Abbvie, Employee of: Galapagos SASU, L. Lepescheux Grant/research support: Abbvie, Employee of: Galapagos SASU, S. Dupont Grant/research support: Abbvie, Employee of: Galapagos SASU, T. Christophe Grant/research support: Abbvie, Employee of: Galapagos NV, M. Borgonovi Grant/research support: Abbvie, Employee of: Galapagos SASU, P. Clément-Lacroix Grant/research support: Abbvie, Employee of: Galapagos SASU, C. Menet Grant/research support: Abbvie, Employee of: Galapagos NV, L. Van Rompaey Grant/research support: Abbvie, Employee of: Galapagos NV, R. Brys Grant/research support: Abbvie, Employee of: Galapagos NV, R. Galien Grant/research support: Abbvie, Employee of: Galapagos SASU DOI 10.1136/annrheumdis-2014-eular.4291

A multiparameter approach to monitor disease activity in collagen-induced arthritis

Introduction Disease severity in collagen-induced arthritis (CIA) is commonly assessed by clinical scoring of paw swelling and histological examination of joints. Although this is an accurate approach, it is also labour-intensive and the application of less invasive and less time-consuming methods is of great interest. However, it is still unclear which of these methods represents the most discriminating measure of disease activity.