IMU-935:Orallyavailablesmallmoleculeinhibitorof IL ... · Result: IMU-935 demonstrated a strong and...
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IMU-935: Orally available small molecule inhibitor of IL-17 with unique molecular profile for the treatment of autoimmune diseases IMU-935 is an orally available RORγt reverse agonist with unique properties: i) synergistic mechanisms of RORγt/DHODH lead to very potent inhibition of the Th17/IL-17 axis, ii) approximately 20% basal RORγt activity at full inhibition, and iii) no effects on thymocyte maturation in vitro. After completion of the IND-enabling studies, IMU-935 will enter phase 1 double-blind, placebo-controlled, single and multiple ascending dose trials in healthy volunteers later this year. We also plan to extend this clinical studies to assess safety and mechanism-related biomarkers in patients with psoriasis. H. Kohlhof 1 , B. Hietel 2 , M. Schenk 2 , M. Gröppel 1 , A. Mühler 1 , Z. He 3 , Z. Sun 3 , S. Tasler 4 , D. Vitt 1 Contact: [email protected] Concept of treating autoimmune diseases by targeting the IL-17 axis The misbalance between pro- and anti-inflammatory signals seems to be the underlaying mechanism in autoimmune and chronic inflammatory diseases. The Th17/IL-17/RORgt axis is thought to be the main player here. Treating patients with antibodies targeting this axis successfully demonstrated this concept but bears the disadvantage of a non-oral drug. With IMU-935 we present an orally available drug targeting this axis and switching back the pro-inflammatory setting to a more balanced immune response. IMU-935 is a unique and potent small molecule inhibitor of Th17 cytokines by targeting RORgT and DHODH Synergistic inhibition of cytokine production 3-5 nM for IL-17A, IL-17F and IFNg à metabolic stress à HEXIM1 expression à Inhibition of transcriptional elongation IM105935 à Inhibition of initiation of target gene transcription IC 50 20 nM DHODH inhibition IC 50 240 nM Method: Inhibition of RORgT was tested in a cellular reporter assay, where LBD of RORg was linked to a GAL4 binding domain. Read-out was luciferase activity. Inhibition of DHODH was tested in a biochemical assay using human DHODH enzyme with DHO as substrate for oxidation, and Decylubiquinon for reduction plus further oxidation of DCIP (Dichlorindophenol) for photometric determination of color changes. Result: IMU-935 is to our knowledge the only RORgT inverse agonist with additional inhibition of human DHODH. This dual inhibition leads to a synergistic and extremely potent inhibition of cytokine secretion. IMU-935 inhibits Th17 differentiation IMU-935 C11, Vitae y = 0.0006x 2 - 0.4443x + 102.25 R² = 0.97064 0 20 40 60 80 100 120 0 100 200 300 400 500 600 Concentration of inhibitor 1 (nM) Normalized T H 17 differentiation (%) 146.6 Method: Murine CD4+ cells were isolated and Th17 differentiation was induced by IL-6, TGFß and IL-23. Cells were treated in parallel with IMU-935 or the Vitae RORgT inverse agonist. The percentage of Th17 cells from viable cells was determined. Result: IMU-935 inhibits the differentiation of murine Th17 cells in a dose dependent manner with an IC50 of around 150 nM. The Vitae RORgT inverse agonist does not display a dose dependent inhibition of Th17 differentiation, even though the in vitro IC 50 levels are similar for IMU-935 and Vitae. No remaining activity of RORgT by Vitae molecule IMU-935 retains basal activity of RORgT and allows normal thymocyte maturation Method: Sorted murine DN thymocytes were cultured on OP9-DL4 fibroblasts with mouse IL-7 for 72h and treated in parallel with IMU-935. Result: IMU-935 allows normal thymocyte maturation from DN towards matured CD4+ thymocytes (CD4+ and CD4+/CD8+). IMU-935 potently inhibited cytokine release in ex vivo stimulated human skin punches 0 20 40 60 80 100 120 140 160 Control Calciportiol 100nM IM105935 100nM IM105935 1µM Control Calciportiol 100nM IM105935 100nM IM105935 1µM Control Calciportiol 100nM IM105935 100nM IM105935 1µM Control Calciportiol 100nM IM105935 100nM IM105935 1µM GM-CSF IL-17A IL-17F IL-22 % of cytokine release compared to control Cytokine Inhibition in Inflamed Skin Model Method: Punches from healthy skin were ex vivo pretreated with IMU-935 for 24h and then challenged with a pro-inflammatory cytokine cocktail for another 24h. Cytokine secretion was measured by ELISA. Result: IMU-935 demonstrated a strong and dose dependent inhibition of GM-CSF, IL-17A, IL-17F and IL-22. Oral application of IMU-935 in IL-17 skin model showed activity Method: Psoriasis like symptoms were induced by topical application of IMQ for 6 days in Balb/c mice. Animals were treated in parallel. Ear score and cytokine expression were determined. Result: Oral application of different doses of IMU-935 leads reduction of ear score and mRNA cytokine levels. IMU-935 demonstrated therapeutic activity in a DSS induced colitis model Method: C57BL/6 mice were challenged with 2.8% DSS from day 0 to day 5 and were treated with IMU-935 from day 4 to day 8. Read-out was diarrhea score, histology with architecture and immune cell infiltration score. Result: Oral application of different doses of IMU-935 improved the symptoms of colitis as depicted by the diarrhea score. Additionally, IMU-935 improved the histological architecture of the gut wall and reduced the infiltration with immune cells. Diarrhea Day 8 Intestinal Architecture Immune Cell Infiltration Colonic TNF⍺ ELISA Gating Strategy for Thymocyte Development RORgT inhibition in cellular reporter assay Inhibition of human RORgT by IMU-935 and Vitae reference compound (C11), compound concentration given in logarithmic scale (10 n x nM). 20% remaining activity of RORgT by IMU-935 Method: Cellular reporter assay for RORgT activity with LBD of RORg fused to GAL4 reporter from Indigo was used. Result: Approximately 20% remaining basal activity of RORgT by IMU-935 at highest dose, whereas Vitae comparator molecule showed full inhibition. 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 unstimulated vehicle stimulated vehicle IMU-935 30mg/kg IMU-935 60mg/kg IMU-935 90mg/kg Dexamethasone IFNgamma mRNA relative expression * ** ** 0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 unstimulated vehicle stimulated vehicle IMU-935 30mg/kg IMU-935 60mg/kg IMU-935 90mg/kg Dexamethasone Ear Score * *** *** 0 0.5 1 1.5 2 2.5 unstimulated vehicle stimulated vehicle IMU-935 30mg/kg IMU-935 60mg/kg IMU-935 90mg/kg Dexamethasone IL-17F mRNA relative expression * * Ear Score IFNg in Skin IL-17F in Skin 1 Immunic AG, Germany, 2 Immunic Research GmbH, Germany, 3 Dept of Molecular Imagine & Therapy, Beckman Research Institute of City of Hope, USA, 4 BioNTech Small Molecules GmbH, Germany CMMI 2019, Trondheim, Norway, June 3-6, 2019
Transcript of IMU-935:Orallyavailablesmallmoleculeinhibitorof IL ... · Result: IMU-935 demonstrated a strong and...
IMU-935: Orally available small molecule inhibitor ofIL-17 with unique molecular profile for thetreatment of autoimmune diseases
IMU-935 is an orally available RORγt reverse agonist with unique properties: i) synergistic mechanisms of RORγt/DHODH lead to very potent inhibition ofthe Th17/IL-17 axis, ii) approximately 20% basal RORγt activity at full inhibition, and iii) no effects on thymocyte maturation in vitro. After completion of theIND-enabling studies, IMU-935 will enter phase 1 double-blind, placebo-controlled, single and multiple ascending dose trials in healthy volunteers later thisyear. We also plan to extend this clinical studies to assess safety and mechanism-related biomarkers in patients with psoriasis.
H. Kohlhof1, B. Hietel2, M. Schenk2, M. Gröppel1, A. Mühler1, Z. He3, Z. Sun3, S. Tasler4, D. Vitt1
Contact: [email protected]
Concept of treating autoimmune diseases by targeting the IL-17 axisThe misbalance between pro- and anti-inflammatory signals seems to be the underlaying mechanism inautoimmune and chronic inflammatory diseases. The Th17/IL-17/RORgt axis is thought to be the main playerhere. Treating patients with antibodies targeting this axis successfully demonstrated this concept but bearsthe disadvantage of a non-oral drug. With IMU-935 we present an orally available drug targeting this axis andswitching back the pro-inflammatory setting to a more balanced immune response.
IMU-935 is a unique and potent small molecule inhibitor of Th17 cytokines by targeting RORgT and DHODH
Synergistic inhibition of cytokine production
3-5 nM for IL-17A, IL-17F and IFNg
à metabolic stressà HEXIM1 expression
à Inhibition of transcriptional
elongation
IM105935
à Inhibition of initiation of target gene
transcription
IC50 20 nM
DHODH inhibitionIC50 240 nM
Method: Inhibition of RORgT was tested in a cellular reporter assay, where LBD of RORg was linked to a GAL4binding domain. Read-out was luciferase activity. Inhibition of DHODH was tested in a biochemical assay usinghuman DHODH enzyme with DHO as substrate for oxidation, and Decylubiquinon for reduction plus furtheroxidation of DCIP (Dichlorindophenol) for photometric determination of color changes.Result: IMU-935 is to our knowledge the only RORgT inverse agonist with additional inhibition of humanDHODH. This dual inhibition leads to a synergistic and extremely potent inhibition of cytokine secretion.
IMU-935 inhibits Th17 differentiation
IMU-935 C11, Vitae
y=0.0006x2 - 0.4443x+102.25R²=0.97064
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Method: Murine CD4+ cells were isolated and Th17 differentiation was induced by IL-6, TGFß and IL-23. Cellswere treated in parallel with IMU-935 or the Vitae RORgT inverse agonist. The percentage of Th17 cells fromviable cells was determined.Result: IMU-935 inhibits the differentiation of murine Th17 cells in a dose dependent manner with an IC50 ofaround 150 nM. The Vitae RORgT inverse agonist does not display a dose dependent inhibition of Th17differentiation, even though the in vitro IC50 levels are similar for IMU-935 and Vitae.
No remaining activity ofRORgT by Vitae molecule
IMU-935 retains basal activity of RORgT and allows normal thymocyte maturation
Method: Sorted murine DN thymocytes werecultured on OP9-DL4 fibroblasts with mouse IL-7 for72h and treated in parallel with IMU-935.Result: IMU-935 allows normal thymocytematuration from DN towards matured CD4+thymocytes (CD4+ and CD4+/CD8+).
IMU-935 potently inhibited cytokine release in ex vivo stimulated human skin punches
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Cytokine Inhibition in Inflamed Skin Model
Method: Punches from healthy skin were ex vivo pretreated with IMU-935 for 24h and then challenged with apro-inflammatory cytokine cocktail for another 24h. Cytokine secretion was measured by ELISA.Result: IMU-935 demonstrated a strong and dose dependent inhibition of GM-CSF, IL-17A, IL-17F and IL-22.
Oral application of IMU-935 in IL-17 skin model showed activity
Method: Psoriasis like symptoms were induced by topical application of IMQ for 6 days in Balb/c mice. Animals were treated in parallel. Ear score and cytokine expression were determined.Result: Oral application of different doses of IMU-935 leads reduction of ear score and mRNA cytokine levels.
IMU-935 demonstrated therapeutic activity in a DSS induced colitis model
Method: C57BL/6 mice were challenged with 2.8% DSS from day 0 to day 5 and were treated with IMU-935from day 4 to day 8. Read-out was diarrhea score, histology with architecture and immune cell infiltration score.Result: Oral application of different doses of IMU-935 improved the symptoms of colitis as depicted by thediarrhea score. Additionally, IMU-935 improved the histological architecture of the gut wall and reduced theinfiltration with immune cells.
Diarrhea Day 8
Intestinal Architecture
Immune Cell Infiltration
Colonic TNF⍺ELISA
Gating Strategy for Thymocyte Development RORgT inhibition in cellular reporter assay
Inhibition of human RORgT by IMU-935 and Vitae referencecompound (C11), compound concentration given in logarithmic scale(10n x nM).
20% remaining activityof RORgT by IMU-935
Method: Cellular reporter assay for RORgT activitywith LBD of RORg fused to GAL4 reporter fromIndigo was used.Result: Approximately 20% remaining basal activityof RORgT by IMU-935 at highest dose, whereasVitae comparator molecule showed full inhibition.
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1Immunic AG, Germany, 2Immunic Research GmbH, Germany, 3Dept of Molecular Imagine & Therapy, Beckman Research Institute of City of Hope, USA,4BioNTech Small Molecules GmbH, Germany
CMMI 2019, Trondheim, Norway, June 3-6, 2019
