Development of Anti-inflammatory Bispecific Trap-antibodies

Abstract

Purpose : A complex interplay of cytokines mediates the pathological responses in retinal inflammatory diseases. Interleukin-1 (IL-1), IL-6, and tumor necrosis factor-alpha (TNF-α) are some of the key regulators of inflammation in the eye. Concurrent inhibition of these proinflammatory cytokines could provide superior efficacy compared to monotherapy. Current treatments which heavily depend on corticosteroids are hampered by the lack of specificity and adverse side effects. Therefore, we engineered specific dual targeting biologics based on trap-antibody fusions against either IL-1 and IL-6, or TNF-α and IL-6.

Methods : The heterodimeric IL-1 receptors were fused to an anti-IL-6 antibody to make an IL-1 trap anti-IL-6 bispecific. Dual binding to both IL-1β and IL-6 was evaluated by surface plasmon resonance (SPR) and mass photometry. Inhibition of IL-1β and IL-6 were assayed in competitive ELISAs. TNF-α trap anti-IL-6 bispecific was developed by fusing the TNF-α receptors to the anti-IL-6 antibody. Dual binding to both TNF-α and IL-6 was evaluated by SPR and mass photometry. Inhibition of TNF-α and IL-6 were assayed in competitive ELISAs.

Results : The IL-1 trap anti-IL-6 bispecific exhibits picomolar affinities to IL-1β and IL-6 and can bind to both targets simultaneously. In competitive ELISAs, the bispecific inhibits IL-1β and IL-6. Similarly, the TNF-α trap anti-IL-6 bispecific tightly and simultaneously binds to both TNF-α and IL-6 and inhibits both cytokines in competitive ELISAs.

Conclusions : This work highlights the expansion of our modular trap-antibody platform to include novel anti-inflammatory biologics. We have developed potent bispecifics against proinflammatory cytokines by fusing the extracellular domains of either IL-1 or TNF-α receptors to an anti-IL-6 antibody. These trap-antibodies are promising therapeutics to mitigate the complex effects of ocular inflammation in a controlled yet multi-specific manner.

This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.