IMO-8400, a novel TLR7, TLR8 and TLR9 antagonist, inhibits disease development in mouse models of psoriasis Weiwen e Ja Jiang, Fu-Gang Zhu, Dong Yu, Ekambar R. Kandimalla, a a, Nicola La Monica, and Sudhir Agrawal Idera Pharmaceuticals, Inc, 167 Sidney Street, Cambridge, MA 02139, USA Poster Board Number: P1537 Session: Novel and Cellular Approaches to Autoimmunity Date and Time: May 6, 2012 from 2:30 PM to 3:30 PM Location: Exhibit Hall (Hynes Convention Center) IMMUNOLOGY 2012, May 4-8, 2012 Boston, Massachusetts
Introduction Tlllik Toll-like Receptosr (TLRs) 7, 8 and 9 can recognize endogenous immune complexes containing self-rna and -DNA, respectively. In many autoimmune diseases, TLR7- and 9-mediated inflammation induced by immune complexes leads to maintenance and progression of disease. Activation of TLR7, TLR8 and TLR9 in pdc and mdc through the interaction of these receptors with the antimicrobial peptide LL37 complexed with self-rna or DNA contributes to psoriasis development 12 1,2. Therefore, inhibition of TLRs 7, 8 and 9 through the use of a TLR antagonist could provide therapeutic effect in this autoimmune disease. We have identified a first-in-class DNA-based antagonist of TLR7, 8, and 9, referred to as IMO-8400. IMO-8400 inhibitstlr7- and 9-mediated immune responses in mice and TLR7-, 8- and 9-mediated immune responses in human cell-based assays and in non-human primates. In the present study, we have evaluated IMO-8400 as a therapeutic agent in IL-23- and LL-37-induced psoriasis models in mice. 1 Lande R, et al., Plasmacytoid dendritic cells sense self-dna coupled with antimicrobial peptide. Nature 449: 564-569, 2007 2 Ganguly D, et al., Self-RNA-antimicrobial peptide complexes activate human dendritic cells through TLR7 and TLR8. J. Exp. Med. 206: 1983-1994, 2009 2011, Idera Pharmaceuticals www.iderapharma.com 2
Role of TLRs 7, 8 and 9 in Psoriasis : Induction of IL-23 and IL-17 Pathways Protein-RNA/DNA complex TLR7/9 pdc IFN-α TLR8 IMO-8400 TLRs 7, 8 and 9 antagonist CD11c+ DC IL-23 IL-20 IL-17 KC activation B-Defensin S100A7 IL-8 IL-22 Th17 IL-12 Th1 IFN-γ TNF-α KC activation MIG IP-10 2011, Idera Pharmaceuticals www.iderapharma.com 3
Outline of Study Protocol of IL-23-Induced Psoriasis Day 0 1 2 3 4 5 6 IL-23, 1 μg, i.d. once a day IMO-8400, s.c. once a day Termination Female C57BL/6 mice, 6 weeks old Experimental groups (n = 8/group) IMO-8400, 300 μg/dose (15 mg/kg) PBS Naïve Evaluation Skin histology Gene expression in skin 2011, Idera Pharmaceuticals www.iderapharma.com 4
Treatment with IMO-8400 Suppresses Skin Lesions Induced by IL-23 PBS-treated t IMO-8400-treated t IL-23 induced various degrees of erythema and induration. IMO-8400 inhibited inflammation induced by IL-23 2011, Idera Pharmaceuticals www.iderapharma.com 5
Treatment with IMO-8400 Inhibits Skin Inflammation Induced by IL-23 Naive PBS-treated IMO-8400-treated Epidermal hyperplasia Inflammatory cell infiltration Abscess HE stain, Magnification x 200 2011, Idera Pharmaceuticals www.iderapharma.com 6
Treatment with IMO-8400 Impacts Gene Expression in IL-23-Injected Skin S100A DEFB4 IL-10 180 2100 12 Fold change 120 60 Fold Change 1400 700 Fold Change 8 4 0 0 Naive PBS IMO-8400 Naive PBS IMO-8400 0 Naive PBS IMO-8400 P < 0.05 vs PBS group Expression of S100 and β-defensin (DEFB4) genes is up-regulated in lesions of psoriasis patients; IMO-8400 significantly suppressed expression of both genes. Gene expression was determined by quantitative real-time PCR 2011, Idera Pharmaceuticals www.iderapharma.com 7
Treatment with IMO-8400 Reduces IL-17 and Increases IL-10 Levels in IL-23-Injected Skin IL-17 IL-10 40 25 30 20 ein pg/mg prot 20 ein pg/mg prot 15 10 10 5 0 0 Naive PBS IMO-8400 Naive PBS IMO-8400 P < 0.05 vs PBS group. Cytokine levels were determined by ELISA 2011, Idera Pharmaceuticals www.iderapharma.com 8
Splenocytes From IMO-8400-Treated Mice Show Reduced Response to TLR9 Agonist Stimulation IL-6 IL-12 P < 0.05 vs PBS. Cytokine levels were determined by ELISA. 2011, Idera Pharmaceuticals www.iderapharma.com 9
Outline of Study Protocol of LL-37-Induced Psoriasis LL-37, 50 μg, i.d. at root of ear once every two days Day 0 4 7 13 16 17 18 IMO-8400, s.c. once every two days Termination Female C57BL/6 mice, 8 weeks old Experimental groups (n = 7/group) IMO-8400, 50 μg/dose (2.5 mg/kg) IMO-8400, 100 μg/dose (5 mg/kg) IMO-8400 8400, 300 μg/dose (15 mg/kg) PBS Naïve Evaluation Ear thickness Skin histology 2011, Idera Pharmaceuticals www.iderapharma.com 10
Treatment with IMO-8400 Suppresses Ear Thickness Increase Induced by LL-37 550 PBS Ear th hickness, μm 450 350 50 μg 100 μg 300 μg IMO-8400 Treatment start day 250 0 2 4 6 8 10 12 14 16 18 19 Days P < 0.05 vs. PBS group 2011, Idera Pharmaceuticals www.iderapharma.com 11
Treatment with IMO-8400 Reduces Skin Inflammation Induced by LL-37 Naive PBS IMO-8400, 300 μg Epidermal hyperplasia p Inflammatory cell infiltration HE stain, Magnification x 100 12 2011, Idera Pharmaceuticals www.iderapharma.com
Summary IMO-8400 is a first-in-class antagonist of TLR7, 8 and 9 In IL-23-induced psoriasis model, IMO-8400 suppressed Psoriatic lesions, epidermal hyperplasia and inflammatory cell infiltration DEFB4 and S100A gene expression and IL-17 protein levels TLR9-mediated immune responses in splenocytes IMO-8400 treatment increased IL-10 gene expression and protein levels in skin In LL-37-induced d psoriasis i model, IMO-8400 reduced d Ear thickness, epidermal hyperplasia and inflammatory cell infiltration IMO-8400 is in development for treatment of autoimmune diseases 2011, Idera Pharmaceuticals www.iderapharma.com 13