Background/Purpose

The therapeutic options for treatment of rheumatic diseases have grown and now include a variety of inflammatory pathway inhibitors, with diverse mechanisms, but having both shared and unique side effects.  Understanding side effect mechanisms is important for guiding patient treatment choices and also for developing improved next generation therapies.   We have successfully employed high throughput primary human cell-based models of tissue and disease, BioMAP Systems, to study failed and approved drugs and have previously identified in vitro activities that correlate with certain side effects.   Here we compare the activity profiles of adalimumab (TNF inhibitor), MTX and anakinra (IL1RA) to test if activities can be correlated with differences in efficacy or safety.

Methods

BioMAP systems model disease biology in early-passage human primary cells and have been used extensively to characterize compounds based on phenotypic signatures.  A panel of 12 BioMAP systems, including mono- and co-cultures of vascular, immune, and tissue cell types were used to generate profiles of anakinra, adalimumab, and MTX.  Changes in protein-based and clinically relevant endpoints (biomarkers, including inflammatory, immune, tissue remodeling and hemostasis-related endpoints) as well as other cellular events (e.g., proliferation, cell cytotoxicity) were evaluated.  For select activities, a comparison to a large reference database of approved and failed drugs, experimental chemicals, and other agents, was performed to elucidate potential mechanisms.

Results

The profiles for both anakinra and adalimumab show anti-inflammatory activities across the panel of BioMAP systems, including reduction in leukocyte recruitment molecules IL-8, E-selectin and MCP-1.  Anakinra was more effective in blocking responses in a co-culture system of monocyte-driven (TLR4) vascular inflammation (LPS system) and differentially active in a model of T cell-dependent B cell activation (BT system), reducing IL-17A, IL-17F, and IL-6.   In contrast, adalimumab was more effective in co-cultures driven by T cell or macrophage (TLR2) activation (SAg and /Mphg systems).  The profile for MTX was distinct, reducing T cell proliferation (SAg system) and IgG production (BT system).  Interestingly, in a fibroblast model of wound healing (HDF3CGF system), anakinra, but not adalimumab or methotrexate increased the levels of VCAM-1 and I-TAC (CXCL11).  VCAM-1 and I-TAC (CXCL11) mediate recruitment of inflammatory lymphocytes into sites of inflammation. This combination of activities is an unusual feature, shared by MEK and p38 MAPK inhibitors, that we have previously associated with the potential for skin rash side effect. 

Conclusion

Profiling of anakinra across a panel of primary human cell systems reveals an activity signature that has been correlated with skin side effects, and may be related to the cutaneous side effects observed with anakinra in patients. This signature is shared by inhibitors of MEK and p38 MAPK and suggests a common pathway mechanism.   A better understanding of side effect mechanisms can help in the design and selection of novel therapies, or combinations.

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« Back to 2014 ACR/ARHP Annual Meeting

ACR Meeting Abstracts - https://acrabstracts.org/abstract/analysis-of-anakinra-in-primary-human-cell-systems-reveals-an-in-vitro-signature-for-skin-related-side-effects/