Piet Wigerinck

Powder for reconstitution of the anti-HIV-1 drug TMC278 - Formulation development, stability and animal studies

Powders for reconstitution of the next-generation non-nucleoside reverse transcriptase inhibitor (NNRTI) TMC278 with low water solubility were developed by using a spray-dry technology. Their flexible dosing ability makes them suitable for patients looking for a different approach for antiretroviral (ARV) therapy. The selection of formulation excipients was based on their potential to create and maintain supersaturation solubility of TMC278 in 0.01 M HCl. Suitable water-soluble carriers for TMC278 were selected by a supersaturation screening to formulate powders for reconstitution by spray-drying. The selected powders for reconstitution were compared to clinical tablets of TMC278.HCl, in vitro using dissolution and stability testing, and in vivo through administration to beagle dogs, fed immediately after dosing. The spray-dried powders for reconstitution made up of TMC278/PVP-VA 64 1:9 (w/w) and TMC278/PVP-VA 64/Cremophor EL 1:8.5:0.5 (w/w/w) showed ease of suspendability, nearly complete dissolution of the drug and acceptable stability after one month storage at 25 and 40 degrees C. In dogs, TMC278 was more slowly absorbed from tablets than from the suspended powders for reconstitution. Compared to the tablet, the relative bioavailability obtained with the powders ranged between 69% and 89% for TMC278/PVP-VA 64 1:9 (w/w) and between 85% and 157% for TMC278/PVP-VA 64/Cremophor EL 1:8.5:0.5 (w/w/w). The absence of differences in vivo and in vitro between the powders made an eventual choice very difficult, yet their advantageous in vivo behaviour and flexible dosing possibility may provide a starting point for paediatric formulations.

OP0263 Efficacy and safety of GLPG0634, a selective JAK1 inhibitor, after short-term treatment of rheumatoid arthritis; results of a phase IIA trial

Background GLPG0634 is an orally-available, selective inhibitor of Janus kinase 1 (JAK1). JAKs are critical components in signaling pathways for a number of cytokines and growth factors, including those involved in the disease process of rheumatoid arthritis (RA). Other non-selective JAK inhibitors have shown long-term efficacy in RA trials with an early onset of action, but doses and thereby efficacy are limited by side effects. By specifically targeting JAK1, treatment with GLPG0634 is expected to result in a cleaner safety profile while maintaining the rapid onset of clinical efficacy. Objectives Evaluate the short-term efficacy and safety of GLPG0634 in RA patients with insufficient response to methotrexate (MTX) alone. Methods A double-blind, placebo-controlled Proof-of-Concept trial in patients with active RA, showing an insufficient response to MTX, was conducted. Twenty-four patients with moderate to severe disease received GLPG0634 200 mg daily, half as once-daily regimen (200 mg q.d.) and half as twice-daily regimen (100 mg b.i.d.), and 12 patients received a matching placebo for a period of four weeks, while continuing to take their stable background therapy of MTX. Results Patient and disease characteristics were comparable for all three dose groups, with DAS28 (CRP) disease activity scores between 6.3 and 6.6. In each dose group, 11 out of 12 patients were female. The patients were well exposed to GLPG0634, with the same pharmacokinetics as previously observed in healthy volunteers. GLPG0634 was found well tolerated and safe with a rapid onset and high level of efficacy. Considering the small sample size, no clinically relevant differences were observed among the 100 mg b.i.d. and the 200 mg q.d. dose regimens. GLPG0634 met the primary endpoint of significant improvement in ACR20 response rate. Overall ACR20 responses were observed in 83% of patients receiving GLPG0634 vs. 33% of patients receiving placebo (p<0.01). GLPG0634 showed impressive results in secondary efficacy endpoints, such as the DAS28 (-2.5), and in reductions of serum C-reactive protein levels (-24.4 mg/L), both significant changes vs. placebo (p<0.0001). All patients completed the trial and no treatment-emergent safety signals were reported. No severe adverse events were reported in patients receiving GLPG0634. Instead of anemia, a modest improvement in hemoglobin was observed. In contrast to observations with JAK inhibitors with other selectivity profiles, no increases in LDL/cholesterol and no liver effects (ALT/AST) were observed in this trial. Conclusions These early clinical results are the first to demonstrate that selective inhibition of JAK1 is efficacious and safe for the treatment of RA. Consistent with the lack of inhibition of JAK2, no anemia was observed. Remarkable results include the high level of response within 4 weeks and the absence of effects on LDL/lipid. An extended GLPG0634 dose-range finding trial is now being conducted to further define the optimal doses for efficacy and safety for longer term studies. Disclosure of Interest F. Vanhoutte Employee of: Galapagos, M. Mazur: None Declared, F. Namour Employee of: Galapagos, A. van der Aa Employee of: Galapagos, P. Wigerinck Employee of: Galapagos, G. Van ’t Klooster Employee of: Galapagos

Oral administration of GLPG0259, an inhibitor of MAPKAPK5, a new target for the treatment of rheumatoid arthritis: a phase II, randomised, double-blind, placebo-controlled, multicentre trial

Background Mitogen-activated protein (MAP) kinases are key regulators of cytokine production, and are therefore potential targets for treatment of rheumatoid arthritis (RA). Objective This two-part phase II study investigated the efficacy and safety of a once-daily 50 mg GLPG0259 (an inhibitor of MAP kinase-activated protein kinase 5) dose vs placebo (part A). An interim analysis after part A would determine whether the dose-finding part (part B) would be performed. Methods In part A, eligible methotrexate (MTX)-refractory patients with RA were randomised to receive either a once-daily 50 mg dose of GLPG0259 or placebo, in addition to a stable dose of MTX, for 12 weeks. The primary efficacy end point was the percentage of patients achieving an American College of Rheumatology 20% improvement (ACR20) response after 12 weeks. Results The interim analysis showed no difference between the percentage of subjects achieving the primary efficacy variable of ACR20 or the secondary efficacy variables (ACR50, ACR70 and Disease Activity Score 28) at week 12 in the GLPG0259-treated (n=19) and placebo-treated (n=11) groups. Owing to lack of efficacy, the study was terminated, and part B was not initiated. Conclusions This innovative study design quickly provided conclusive results on the lack of efficacy of GLPG0259 in patients with RA.

Pharmacokinetics, Safety, and Tolerability of GLPG0259, a Mitogen-Activated Protein Kinase-Activated Protein Kinase 5 (MAPKAPK5) Inhibitor, Given as Single and Multiple Doses to Healthy Male Subjects

Background and ObjectivesGLPG0259 is a small-molecule inhibitor of mitogen-activated protein kinase-activated protein kinase 5 (MAPKAPK5), a kinase enzyme that plays a role in important inflammatory pathways. The main objectives of the phase I clinical studies in early development were to characterize the pharmacokinetics, tolerability, and safety of GLPG0259 in healthy subjects, including the development of a solid dosage form (free-base pellets and fumarate salt capsules) and the potential for interaction of GLPG0259 with methotrexate.Subjects and MethodsFour phase I studies were initiated. Study 1 was a randomized, double-blind, placebo-controlled study to evaluate the safety, tolerability, and pharmacokinetics of single ascending doses (1.5–150 mg) and multiple oral doses (20 and 50 mg once daily) of GLPG0259 in healthy male subjects (n = 34). Study 2 was a randomized, double-blind, placebo-controlled study to evaluate the safety, tolerability, and pharmacokinetics of oral multiple ascending doses of GLPG0259 (25–75 mg once daily) given for 14 days to healthy male subjects, and to get preliminary information on the potential pharmacokinetic interaction between GLPG0259 and methotrexate (n = 24). Studies 3 and 4 were open-label, randomized, crossover studies to compare the oral bioavailability of two solid dosage forms of GLPG0259 (a capsule) relative to an oral solution after a 100 mg or 50 mg single dose and to evaluate the effect of food on these formulations (n = 12 for each study).Main Outcome MeasuresThe non-compartmental pharmacokinetic parameters for plasma concentrations of GLPG0259 were determined, and a population pharmacokinetic model of GLPG0259 was developed to support the planning of the number and timing of the sparse samples to be taken per patient in the phase II study. Safety and tolerability data are also summarized.ResultsThe absorption of GLPG0259 was slow, with a decrease in the absorption rate with increasing dose, and there was decreased elimination, with an apparent terminal elimination half-life of 26.0 hours. On the basis of statistical analysis of variance, the exposure to GLPG0259 increased in proportion to the dose over a 30–150 mg single-dose range and a 25–75 mg repeated-dose range. Between- and within-subject variability in GLPG0259 pharmacokinetics was low/moderate (coefficient of variation [CV] 16–30%). After once-daily repeated dosing, steady-state plasma concentrations were reached at between 5 and 8 dosing days, which is consistent with the long apparent elimination half-life of GLPG0259. Food increased the bioavailability of GLPG0259 given in a solid dosage form. Co-administration of GLPG0259 with a single dose of methotrexate 7.5 mg did not result in any change in the pharmacokinetic profiles of either GLPG0259 or methotrexate.ConclusionIn summary, the investigation of safety/tolerability and pharmacokinetics in the early development phase showed that single and repeated doses of GLPG0259 were safe and well tolerated. The most common adverse event reported was mild gastrointestinal discomfort. The pharmacokinetics characterized in healthy male subjects showed no major obstacles and supports a once-daily oral regimen in patients.

Abstract 1568: GLPG0187, a small molecule integrin antagonist, shows good safety and decrease in CTX levels in single ascending dose study

Introduction : GLPG0187, a small molecule integrin antagonist with nanomolar affinity for the RGD-integrin receptors αvβ1, αvβ3, αvβ5, αvβ6 and α5β1, shows potent anti-angiogenic, anti-metastatic, anti-bone-resorption and anti-tumor activity in vitro and in vivo. Therefore GLPG0187 may offer a promising therapeutic approach for the treatment of integrin receptor expressing tumors and/or metastases. Methods: Based on promising preclinical data, a first-in-human study was performed with GLPG0187 to assess the safety and tolerability as well as pharmacokinetic and pharmacodynamic behavior of single ascending doses of the compound. GLPG0187 was administered subcutaneously in a dose range of 17.5 to 315 mg and orally in a dose range of 50 to 1200 mg. Results: GLPG0187 was systemically well tolerated with no adverse effects on ECG, vital signs or laboratory parameters. GLPG0187 showed a dose proportional pharmacokinetic profile over the dose range tested. Moreover, GLPG0187 dose proportionally decreased osteoclast activity as measured by plasma CTX (carboxy-terminal collagen crosslinks) levels. Conclusions: GLPG0187 was shown to be safe and well tolerated, to have a dose proportional pharmacokinetics and to inhibit osteoclast activity. These results provide further evidence that GLPG0187 could be an effective therapeutic for the treatment of cancer. Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1568.

Abstract 1753: GLPG1790: The first Ephrin (EPH) receptor tyrosine kinase inhibitor for the treatment of triple negative breast cancer

Receptor tyrosine kinases define a clinically relevant class of targets in the field of cancer. Here we report the discovery of a pre-clinical drug candidate directed against the EPH receptor family. Members of this family of receptor tyrosine kinases are over-expressed in diverse cancer types. GLPG1790 is a small molecule, nanomolar inhibitor of various EPH receptors kinases. The compound displayed a remarkable efficacy by once-daily oral administration in a mouse EPHA2 expressing xenograft model (MDA-MB-231). A rapid dose-dependent reduction of tumor growth was achieved, with full inhibition at the oral dose of 30 mg/kg/d. GLPG1790 efficacy at this dose was similar to that of Paclitaxel administered at its maximum tolerated dose. A series of experiments was initiated to confirm the mechanism of action of this compound. GLPG1790 inhibits human EPHA2 kinase activity with an IC 50 of 11 nM in a biochemical assay. In the human MDA-MB-231 breast cancer cell line, that expresses a high level of EPHA2 protein, GLPG1790 inhibits receptor phosphorylation with an IC 50 of 260 nM, and anchorage-independent growth with similar potency. In addition, in vivo target engagement was demonstrated in the mouse MDA-MB-231 xenograft model. GLPG1790 inhibited EPHA2 receptor phosphorylation after single oral administration of 30 and 100 mg/kg doses. The extent of the effects observed on EPHA2 phosphorylation correlated with intra-tumoral GLPG1790 concentration. Moreover, the MAPK pathway, known to be a major driver of proliferation of this cell line, was inhibited both in in vitro cellular assays and in xenograft target engagement studies. Flow cytometric analyses revealed a cell cycle arrest at the G0/G1 phase for MDA-MB-231 cells treated with GLPG1790. EPHA2 knock-down-based experiments further support the EPH-driven mode of action of GLPG1790. All together these data stimulate the development of GLPG1790 in triple negative breast cancer. This novel mechanism of action is under investigation in other cancer types overexpressing EPH9s (melanoma, pancreatic, ovarian, prostatic and colorectal cancers). Citation Format: Philippe Pujuguet, Filip Beirinckx, Carole Delachaume, Jacques Huck, Ellen Van der Aar, Reginald Brys, Luc Van Rompaey, Piet Wigerinck, Laurent Saniere. GLPG1790: The first Ephrin (EPH) receptor tyrosine kinase inhibitor for the treatment of triple negative breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1753. doi:10.1158/1538-7445.AM2014-1753

THU0236 Once-Daily Dosing of GLPG0634, a Selective JAK1 Inhibitor, is Supported by Its Active Metabolite

Background GLPG0634 is an orally-available, selective, inhibitor of Janus kinase 1 (JAK1). Through inhibition of signaling pathways for cytokines involved in rheumatoid arthritis (RA), non-selective JAK inhibitors have shown long-term efficacy in treating RA but also dose-limiting side effects. Selective inhibition of JAK1 may result in an acceptable safety profile while maintaining clinical efficacy and a rapid onset of action. Both once-daily (QD) and twice-daily (BID) dosing regimens have been explored in humans. QD dosing of GLPG0634 in humans has shown highly encouraging pharmacodynamic (PD) properties, and efficacy in RA as demonstrated in a 4-week Proof of Concept study. Objectives Evaluate the PK, PD and efficacy of GLPG0634 and its main metabolite(s) in healthy volunteers and patients with active RA. Methods GLPG0634 was dosed in healthy volunteers (N=36) at doses of 25, 50 and 100 mg BID, and 200, 300 and 450 mg QD for 10 days, and in RA patients (N=24) at 100 mg BID and 200 mg QD for 28 days added on to a background therapy of methotrexate. The clinical PK of GLPG0634 and its active major metabolite was evaluated, and PD in whole blood from healthy volunteers through ex vivo IL-6 induced phosphorylation of STAT1 (pSTAT1) in CD4+ cells was evaluated for JAK1 activity. For JAK2 activity, GM-CSF induced pSTAT5 was measured in CD33+ cells. JAK inhibition and selectivity of the metabolite were evaluated in vitro. Results The PK for GLPG0634 following BID and QD dosing was predictable from prior single dose data, showing dose-proportionality and a half-life of 7 h. Its major active metabolite showed plasma levels in humans well exceeding those of GLPG0634 and a half-life of 22 h. The PK profile in healthy volunteers and RA patients was similar. IL-6 induced pSTAT1 was inhibited for the entire 24 h dosing period, including at 24 h after the last dose for dose regimens of 200 mg QD. No relevant inhibition of JAK2 activity was observed at any dose level. While plasma levels of GLPG0634 were low at these time points, those for the major metabolite remained high. QD dosing showed a high level of efficacy within 4 weeks with 75% of patients achieving ACR20, a decrease in DAS28 (CRP) of 2.2, and normalization within 7 days of the high baseline CRP levels. While the major metabolite was found to be a 10 to 20-fold less potent kinase inhibitor generally, its selectivity forJAK1 was similar to that of GLPG0634. Conclusions The results of these studies indicate that an active metabolite supports the activity of GLPG0634. The long half-life of this metabolite provides a lasting effect, though at a lower level of JAK1 inhibition than GLPG0634. The lower potency for inhibition of JAK1 is, at least in part, compensated by the high exposure in humans. These findings may explain the sustained efficacy results observed with QD dosing of GLPG0634, and the 24-hour suppression of JAK1 in whole blood from healthy volunteers on GLPG0634, in spite of its moderate half-life. These data indicate that a major active metabolite supports longer lasting effects of GLPG0634 and reduces fluctuations in JAK1 inhibition. This may allow for QD dosing in a therapeutic setting, adding to dosing convenience for patients. Disclosure of Interest F. Namour Employee of: Galapagos SASU, R. Galien Employee of: Galapagos SASU, F. Vanhoutte Employee of: Galapagos NV, P. Wigerinck Employee of: Galapagos NV, G. van ’t Klooster Employee of: Galapagos NV