M. L. Faquetti, F. Grisoni, P. Schneider, G. Schneider, A. M. Burden
2021 Annals of the Rheumatic Diseases  
Background:The JAK inhibitors (JAKi's) tofacitinib and baricitinib are new alternatives for treating rheumatoid arthritis. Safety concerns associated with JAKi's, such as the increased risk for thrombosis and viral infections, have emerged worldwide. The underlying explanatory mechanisms remain unknown, suggesting the elevated risk is likely due to underlying confounding or an off-target binding effect. Computational approaches can explore the potential for a small molecule drug to interact
more » ... previously unknown biological targets and identify potential safety-related concerns, and open doors for potential drug repurposing.Objectives:To identify and characterize the off-target binding effects of baricitinib and tofacitinib, with a focus on targets related to thrombosis and viral infectionMethods:Potential targets of baricitinib and tofacitinib were predicted using two neural-network-based systems (TIGER[1] and SPiDER[2]). Targets were considered relevant if they had (1) a SPiDER confidence with p<0.05, or (2) a TIGER score >1. Selected targets related to the outcome of interest were experimentally evaluated at Eurofins Cerep (France-Celle L'Evescault, www.eurofins.com) if commercial available. Compounds were tested at (1) single concentration (30 µM) with technical replicates, using radioligand or enzymatic assays, or (2) multiple concentrations (30 µM highest concentration; dilution factor in a log-scale) with technical replicates, using calcium flux or inhibition of [cAMP] assays. Observed activity of ≥50% inhibition or stimulation on the target was considered active, between 25 to 50% inhibition (or a dissociation constant [Kd] from 1 to 10 µM) was considered as moderate activity, and lower than 25% was considered inactive. Dose-response curve were performed on active and moderate targets for IC50 / EC50 (half maximal inhibitory / effective concentration) determination.Results:TIGER and SPiDER suggested a total of 99 off-target binding effects (baricitinib n=41; tofacitinib n=58), of which 17 targets had potential impact on thrombosis or viral infection (baricitinib n=5 and 4, respectively; tofacitinib n=5 and 3, respectively). Commercial testing was available on 11 targets (Adenosine Receptor A2A [AA2AR], Epidermal growth factor receptor, induclible NOS, PI3 Kinase (p110b/p85a), Phosphodiesterase 10A2 [PDE10A2] and Protein Kinase N2 [PKN2] for baricitinib; and Adenosine receptor A3, 15-Lipoxygenase [15-LO], PKN2, Transient receptor potential cation channel [TRPM6] and AA2AR for tofacitinib). Of these, 5 targets showed active or moderately active binding activity (baricitinib n=2; tofacitinib n=3), and were tested for dose-response curves. Test results confirmed ligand-binding activity with IC50 on nanomolar (PKN2), and micromolar ranges (PDE10A2 and TRPM6).Conclusion:The results suggest both baricitinib and tofacitinib are promiscuous binders with effects on several families. Although it may lead to side effects, off-target binding also represents a potential opportunity for drug repurposing. Besides on-target effects, both drugs are under clinical investigation for the treatment of COVID-19 due to off-target interactions. The proposed pharmacological off-target effects of those with active binding include attenuation of pulmonary vascular remodeling, anti-fibrotic and anti-psychotic activities (PDE10A2), modulation of viral response (PKN2), and hypomagnesaemia (TRPM6), which is involved in cardiovascular diseases. This study supports tofacitinib and baricitinib as candidates for drug repurposing (e.g., in COVID-19, Hepatitis C virus, and pulmonary hypertension). We did not identify active off-target interactions linked to thrombosis to explain the elevated risk observed in clinical practice. Further research is required to elucidate the underlying patient-specific factors (confounders) that could explain this safety concern.References:[1]Schneider P et al. Angew Chem Int Ed 2017;56:11520–4.[2]Reker D et al. PNAS 2014;111:4067–72.Disclosure of Interests:None declared
doi:10.1136/annrheumdis-2021-eular.982 fatcat:o7imgfn5efhbrmjdzndp26royu