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Results
Effects of TOP1210, a narrow spectrum kinase inhibitor, and selective
kinase inhibitors on the intestinal pro-inflammatory immune response in
ulcerative colitis
Martyn R Foster‡, Paolo Biancheri†, Matthew CT Fyfe, Thomas T MacDonald †, Adele Rowley‡, Sameer Sirohi, Yemisi Solanke, Steve
Webber‡, Eleanor Wood♯ and Claire A Walshe.‡
‡Topivert Pharma Ltd., London, UK; †Centre for Immunobiology, Barts and the London School of Medicine and Dentistry, London, UK; ♯Academic Department of Medical and
Surgical Gastroenterology, Homerton University Hospital, London, UK.
Figure 3. TOP1210 inhibitory effects on pro-inflammatory cytokine release from UC biopsies.
Introductio
n
Geometric Mean IC50 value (nM)
p38α Src Syk
TOP1210 65 10 17
BIRB-796 13 >1895 >1895
Dasatinib 378 6 >2049
BAY-61-3606 >2562 >2562 136
Geometric Mean IC50 value (nM)
Innate response: Monocytes Adaptive response: T
cells Epithelium
PBMCs Primary macrophages PBMCs PBMCs HT29 cells
IL-8
release IL-8 release
TNF-
release
IL-2
release
IFN-
release
IL-8
release
TOP1210 1.9 2.2 3.3 37 2.1 1.8
BIRB-796 >1895 >250 >474 >1895 >1895 >1895
Dasatinib >2049 >250 52 2.1 4.0 >2049
BAY-61-
3606 607 >250 >640 291 247 >2561
Table 2 : Effect of TOP1210 and selective kinase inhibitors on a
range of innate, adaptive and epithelial cellular response assays.
Table 1: Inhibitory effects of selective kinase inhibitors and the
NSKI TOP1210 on kinase activity in a biochemical Z-lyte based
assay.
Table 3. Inhibitory effects of the
NSKI TOP1210 and the selective
kinase inhibitors on pro-
inflammatory cytokine release by
Ulcerative Colitis myofibroblasts.
Figure 1. Exemplar data from cellular assays. TOP1210 inhibits
IL-8 release by LPS stimulated PBMCs (Figure 1A) and
macrophages (Figure 1B), and also LPS stimulated TNFα release
by macrophages (Figure 1C), with greater potency and efficacy
than any of the selective kinases tested.
• TOP1210 potently inhibits all kinases tested, whilst BIRB-
796, dasatinib and BAY-61-3606 show selective profiles.
IL-6
(IC50 ng ml-1)
IL-8
(IC50 ng ml-1)
TOP1210 2.2 2.1
BIRB-796 >1000 >1000
Dasatinib >1000 >1000
BAY-61-
3606
315 630
• Intracellular kinase activation plays a key role in inflammation and kinase inhibitors have been proposed as potential therapies in chronic inflammatory disorders such as ulcerative colitis.
• Selective kinase inhibitors, however, have proved disappointing, particularly in the treatment of rheumatoid arthritis and inflammatory bowel disease (IBD).
• Multi-kinase inhibition has been investigated as a strategy to improve efficacy.
• The activity of a narrow spectrum kinase inhibitor (NSKI), TOP1210, has been compared to selective kinase inhibitors (BIRB-796, dasatinib and BAY-61-3606) in a range of innate and adaptive inflammatory cell assays and in inflamed biopsies from ulcerative colitis (UC) patients.
Methods • Inhibitory effects on recombinant p38, Src and Syk kinases was
assessed in an ATP dependent ZYLTE™ based assay.
• Cellular assays were performed after 2hr preincubation with compound or vehicle. Peripheral blood monocytes (PBMCs) were stimulated with either lipopolysaccharide (LPS) or anti-CD3/CD28, monocyte derived macrophages with LPS, HT29 epithelial cells with IL1β and myofibroblasts isolated from inflamed UC mucosa with TNFα.
• Biopsies from inflamed ulcerative colitis patients were incubated with compound (24hr) and spontaneous cytokine release measured.
• A range of inflammatory cytokines in cellular and biopsy supernatants were measured by ELISA. Data is expressed as means ±s.e.m.of at least 3 determinations.
Conclusions
• Multi-kinase inhibition with NSKIs like TOP1210 leads to a efficacious and broad inhibitory profile in UC tissues and across a range of cell types including epithelial cells, innate and adaptive immune cells. Thus, NSKIs provide significant advantages over existing selective kinase approaches, and potentially offer much improved therapeutic benefit in IBD.
Results Introduction
Figure 2. Effect of TOP1210,
BIRB796 (BIRB), dasatinib
(DAS), or BAY-61-3606 (BAY)
on the release of IL-6 and IL-8
by UC myofibroblasts
• TOP1210 is more potent than selective kinase inhibitors in
reducing cytokine release from TNFα-stimulated UC
mucosal myofibroblasts.
IL-1 IL-6 IL-8
0 1 0.1 0.01 0.001
TOP1210 (g/ml)
0 1 0.1 0.01 0.001
TOP1210 (g/ml)
0 1 0.1 0.01 0.001
TOP1210 (g/ml)
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• TOP1210 is a potent and efficacious inhibitor in down-
regulating pro-inflammatory cytokine release from Ulcerative
Colitis biopsies.
• TOP1210 is a potent and efficacious inhibitor of innate and
adaptive immune responses. Generally, compared to
TOP1210, the selective kinase inhibitors have weak potency
and efficacy