Canakinumab (Ilaris®) reduces IL-1β-induced IL-6 production by human muscle cells


Pre-Clinical Research

Poster Number: S16


Anna Cordeiro-Santanach, PhD, AGADA Biosciences Inc., Fiorella Morales, AGADA Biosciences, Inc., Pia Elustondo, PhD, AGADA Biosciences Inc., Kanneboyina Nagaraju, PhD, Binghamton University

DMD is characterized by muscle inflammation due to higher levels of pro-inflammatory cytokines such as interleukin 1β (IL-1β). We have assessed the effect of a human IgG1κ monoclonal antibody (canakinumab (Ilaris®)) against IL-1β on the ability of human muscle cells to secrete the pro-inflammatory myokine IL-6.
Canakinumab is a strong candidate for therapeutic repurposing to treat DMD because it is already FDA approved for juvenile and adult autoinflammatory diseases. Unlike other IL-1 inhibitors, canakinumab is highly specific for the IL-1β ligand, has a longer half-life, and does not interfere with other IL-1-activated pathways. Following cell culture optimization and viability assays to assess toxicity, myoblasts were stimulated with IL-1β (10 ng/ml) for 48 hours in the presence of eight concentrations of canakinumab and IL-6 production was measured with an enzyme-linked immunosorbent assay. Incubation of canakinumab with IL-1β-stimulated myoblasts significantly reduced IL-6 production at concentrations of 1, 10, 100, 250, and 1000 nM relative to controls (p<0.001 for all doses), yielding an IC50 of 0.264 nM. On the other hand, canakinumab that was pre-bound to IL-1β and then administered in combination to myoblasts resulted in a higher inhibitory affinity with the IC50 reducing to 0.126 nM, less than half of the previous method. This study also evaluated the effects of canakinumab in reducing IL-6 production in myotubes, the terminally differentiated and functional muscle cells. The ELISA assay showed significant reduced levels of IL-6 after 30 min IL-1β (10 ng/ml) coincubation with canakinumab (10 nM) (p<0.001). This study reveals that canakinumab is a potent inhibitor of IL-1β-stimulated IL-6 production in muscle cells (myoblasts and myotubes). These results align with previously published pre-clinical work in mdx mice and support further investigation into the clinical utility of repositioning canakinumab for treatment of DMD.