NUE7770 A BET-BD1 selective chemical probe with potent cellular and in vivo anti-inflammatory activity. 14 th Discovery on Target 2016, Boston MA
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1 NUE7770 A BET-BD1 selective chemical probe with potent cellular and in vivo anti-inflammatory activity 14 th Discovery on Target 2016, Boston MA
2 NUEVOLUTION The CHEMETICS lead discovery platform Nuevolution A/S Founded 2001 Located in Copenhagen, Denmark Chemetics platform technology Allowing screen of billions of compounds Internal pipeline Small molecules for oncology, inflammation, and IO 42 employees Oncology Inflammation Immunooncology Slide 2
3 NUEVOLUTION The CHEMETICS lead discovery platform Nuevolution A/S Founded 2001 Located in Copenhagen, Denmark Chemetics platform technology Allowing screen of billions of compounds Internal pipeline Small molecules for oncology, inflammation, and IO 42 employees Oncology Inflammation Immunooncology Slide 3
4 NUEVOLUTION The CHEMETICS lead discovery platform Nuevolution A/S Founded 2001 Located in Copenhagen, Denmark Chemetics platform technology Allowing screen of billions of compounds Internal pipeline Small molecules for oncology, inflammation, and IO 42 employees Oncology Inflammation Immunooncology ~ Fragments 15+ Targets (Screening / Y) 3-5 Libs / Y Screens / Y 10 B+ Templates / Y Slide 4
5 BET BROMODOMAIN INHIBITION Potential disease indications for compounds targeting BET proteins Company Compound (mechanism) Indication Stage of development BRD BET Bromodomains (BRD2,3,4,T) BETs are chromatin readers of AcK marks and master transcriptional regulators Merck/Mitsubishi Tanabe Bristol-Myers Squibb Incyte Pharmaceuticals AbbVie MK-8628 (BRD2/3/4 selective small-molecule inhibitor) BMS (undisclosed BRD small-molecule inhibitor) INCB54329 (undisclosed BRD small-molecule inhibitor) ABBV-075 (undisclosed BRD small-molecule inhibitor) AML Phase 2 Malignant glioma, glioblastoma and anaplastic astrocytoma Phase 2 Solid tumors Phase 2 Solid tumors Phase 1/2 Hematologic cancer Phase 1/2 Cancer Phase 1 Vast literature supporting BET function in multiple diseases of cancer & inflammation Many panbet inhibitors in clinical development (oncology) Emerging clinical toxicity reports of panbet inhibitors - GI, hematology etc. Is clinical toxicity linked to pan-bet activity Will selectivity result in less toxicity? Constellation Pharmaceuticals/Roche FORMA Therapeutics/Celgene Gilead Sciences GSK Daiichi Sankyo Tensha Therapeutics/Roche CPI-0610 (BRD4 smallmolecule inhibitor) FT-1101 (BRD2, BRD3, BRD4, BRDT small-molecule inhibitor) GS-5829 (undisclosed BRD small-molecule inhibitor) GSK (BRD2, BRD3, BRD4, BRDT small-molecule inhibitor) PLX51107 (BRD 4 smallmolecule inhibitor) Ten-010 (BRD2, BRD3, BRD4, BRDT small-molecule inhibitor) Source: Biomedtracker, Pubmed and Patent lens. Acute lymphocytic leukemia (ALL) Phase 1 Multiple myeloma Phase 1 Myelodysplastic syndrome Phase 1 AML Phase 1 Myelodysplastic syndrome Phase 1 Diffuse large B-cell lymphoma- non-hodgkin's lymphoma Phase 1 Solid tumors Phase 1 Solid tumors Phase 1 Cancer Phase 1 Solid tumors Phase 1 Slide 5
6 CHEMETICS SCREENING OF BRD4_1 Chemetics Selection Hit Series Optimization Lead BRD4_1 Screening of 2 libraries 10+ Chemical Series Fragments (2 Selected for optimization) 300 M cpds putative hits In the Chemetics screening process, DNA-encoded small molecule libraries and target protein are subjected to one or more bind-wash-elute cycles followed by barcode sequencing and hit deconvolution We screened 2 libraries for affinity toward BRD4_1 >10 chemical series enriched, >1000 putative hits Slide 6
7 NUE COMPOUNDS SELECTIVELY TARGETING BD1 OF BET PROTEINS Binding domain 1 selectivity is a general feature of NUE compounds High BD1 selectivity seen for all BETs BD1 NUE compounds JQ1 IBET-726 RVX-208 BD2 NUE compounds, IC 50 (BROMOscan ) Reference compounds NUE-1 NUE-2 NUE-3 NUE-4 NUE-5 JQ-1 RVX-208 BRD2_1 (nm) BRD3_1 (nm) BRD4_1 (nm) BRD2_2 (nm) BRD3_2 (nm) BRD4_2 (nm) BRD2 BD1/BD2 sel BRD3 BD1/BD2 sel BRD4 BD1/BD2 sel Slide 7
8 PREPARATIONS FOR IN VIVO POC Biomarker study αcd3 injection αcd3 injection 48h 4h IL-17A Esplugues et al. have shown in mice, that injection of stimulatory CD3 antibody twice (in intervals of 48 hrs) leads to increase in IL-17A plasma levels Mele et al. use this model to show inhibition of Th17 pathway in vivo using bromodomain inhibitor JQ-1 We used this in vivo biomarker model to confirm similar effect by our selective BET inhibitors in our optimization for optimal exposure αcd3 injection Nuevolution BET inhibitor αcd3 injection Esplugues et al; Nature 475 (2012) p514 See also Mele et al; J. Exp. Med. 210 (2013) p h 4h IL-17A Slide 8
9 IN VIVO BIOMARKER STUDY Effect on serum IL-17A concentration NUE-4 shows potent effect on IL-17A serum level Reduction of IL-6 and CXCL1 levels Slide 9
10 NUE7770 SELECTIVE TARGETING OF BET-BD1 Attractive compound properties Slide 10 Properties Bromodomain NUE7770, IC 50 BRD2_1 (µm) BRD3_1 (µm) BRD4_1 (µm) 0.15 BRD2_2 (µm) 7.2 BRD3_2 (µm) 2.9 BRD4_2 (µm) 9.4 BRD2 BD1/BD2 selectivity 57 BRD3 BD1/BD2 selectivity 190 BRD4 BD1/BD2 selectivity 64 MW <375 AlogP <3 mhep T½ (min) >200 mhep Clint (µl/min/10 6 cells) <5.3 Kinetic Solubility (FaSSIF) 95µM mppb (%) 89.5 Oral bioavailability, mouse (%) 29 IV clearance, mouse (ml/min/kg) 11 Half-life, mouse (h) 1.0 MDCK permeability Moderate CYP P450 inhibition (1A2, 2C9, 2C19, 2D6, 3A4) All IC 50 >50µM Potent and BD1 selective Good in vitro and in vivo stability Acceptable solubility and permeability No significant inhibition of non-bet bromodomains No kinase activity observed BET_BD1 BET_BD2
11 BIOMAP PROFILE OF NUE7770 Selective and strong dose-dependent response in lymphocytes 10µM 3.3µM 1.1µM 370nM Venular endothelial cells PBMC Venular EC B-cells PBMC Bronchial epithelial cells, Dermal fibroblasts Bronchial epithelial cells Coronary artery smooth muscle cells Dermal fibroblasts Dermal fibroblasts, Keratinocytes Lung fibroblasts Macrophages, Venular EC Disease context Allergy, Asthma, Autoimmunity, Oncology Slide 11
12 BIOMAP PROFILE OF NUE7770 Selective and strong dose-dependent response in lymphocytes 10µM 3.3µM 1.1µM 370nM Venular endothelial cells PBMC Venular EC B-cells PBMC Bronchial epithelial cells, Dermal fibroblasts Bronchial epithelial cells Coronary artery smooth muscle cells Dermal fibroblasts Dermal fibroblasts, Keratinocytes Lung fibroblasts Macrophages, Venular EC IL2 IL17F IL17A IL6 IgG Disease context Allergy, Asthma, Autoimmunity, Oncology Strong and selective cytokine response in multiple systems BIOMAP activity points towards diseases with T cell-dependent antibody production T H 17/IL-17-dependent pathology RA, MS, Lupus Slide 12
13 SHORT DURATION MECHANISTIC IN VIVO MODEL Effect on CXCL1 production from anti-il-17a stimulation -18h: IL-17A -1h: NUE 0h: IL-17A 2h: Plasma 20h CXCL1 (Gro ) levels CXCL1 (Gro ) levels Purpose Demonstrate whether reduction of CXCL1 occurs upstream or downstream from IL-17 Comparison to mouse IL-17A antibody Model αil-17a -18h NUE compound -1h IL-17A 0h NUE compound dosed 30mpk, BID, po 10mpk, BID, po Slide 13
14 THERAPEUTIC EFFECT IN MOUSE MODEL OF CIA Convincing 30mpk BID Therapeutic setting - treatment starts only after mice have developed initial symptoms of arthritis (AI = 3.3) Treatment with NUE7770 almost completely inhibits disease progression No adverse effects observed (BW, clinical observations) Two comparators included in the study: Xeljanz (a JAK inhibitor approved for human RA) and a neutralizing anti-il-17a antibody Slide 14
15 CONCLUSION AND NEXT STEPS Efficacy and safety/tolerability Lupus in-vivo POC upcoming NUE7770 Probe compound Safety And Tolerability Mouse 2-week Tox study: NUE7770 vs JQ-1 In-life completed, preliminary data: Mortality in JQ-1 group consistent with previously reported studies No mortality in NUE7770 groups up to 300mpk, BID Full study data expected Q4/16 Efficacy - clinical scoring and relevant biomarkers in mouse disease models of autoimmune disease Systemic Lupus Erythematosus Rationale Large unmet need Validated B and T H 17 component Pristane-induced lupus model In-life on-going Full study data expected Q4/16 Collagen-induced arthritis (Antibody/IL17A driven) Strong efficacy of NUE7770 in therapeutic setting Slide 15
16 ACKNOWLEDGEMENT BET Team Library production Advisors Boston University Jimmi Seitzberg Tine Titilola Kronborg Jutta Heim Gerald Denis Visnja Poljak Kristian Klindt Jeanette Wood Gitte Friberg Charlotte Andersen Peter Hirth Margit Hansen Kirstine Rugård Gordon Mills Christina Underwood Eva Kampmann Olsen Paul Workman Berit Tonnesen Aziza Bouhzane Marc Feldmann Søren Nielsen Luigi Stasi Lene Teuber Thomas Franch Mads Nørregaard Alex Gouliaev Parallel Chemistry Mikkel Vestergaard David Hjort Cristina Delgado Michael Godskesen Mark Genovese Ron Marler Slide 16
17 Thank You Please check out poster 148 Nuevolution Presentation 14 th Discovery on Target 2016, Boston MA
18 Thank You Please check out poster 148 Nuevolution Presentation 14 th Discovery on Target 2016, Boston MA
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