OP028 Oral delivery of a new class of non-antibody protein scaffold Nanofitins targeting TNF-alpha shows a strong preventive and curative anti-inflammatory effect in models of inflammatory bowel diseases.
M. Cinier*1, M. Zeisser Labouèbe2, C. Rousseaux3, J.V. Rodriguez4, A. Cunha4, N. Truong Tan3, R. Gurny2, L. Scapozza2, O. Kitten1
1Affilogic, Affilogic, Nantes, France, 2University of Geneva, School of Pharmaceutical Sciences, Geneva, Switzerland, 3Intestinal Biotech Development, Intestinal Biotech Development, Lille, France, 4Instituto de Biologia Experimental Tecnológica, iBET, Oeiras, Portugal
Despite a remarkable efficacy, treatment of inflammatory bowel diseases (IBD) using systemic administration of anti-TNF-alpha antibodies remains associated with serious adverse effects. Oral administration of such therapeutics would benefit from a better targeting to the site of inflammation in the gut while decreasing their systemic exposure and related side effects. To this aim, the SADEL FP7 European project has been developing oral formulation of anti-TNF-alpha Nanofitins (NF), a novel alternative scaffold derived from the sac7d protein found in an extremophilic archeabacterium and stable enough to survive the hostile environment of the gut.
Screening of anti-TNF-alpha NF hits was done in vitro using surface plasmon resonance and in vivo by evaluating their anti-inflammatory effects in preventive mode after intrarectal instillation (10 mg/kg) in TNBS-induced model of colitis (C57Bl6 mice), using 5-ASA at optimal dosage (30 mM) for comparison. NFs providing reduction of inflammation similar or superior to 5-ASA were engaged in a dose-range finding study in the same model, and the optimal dose has been confirmed in DSS-induced model of colitis (Balb/C mice). Oral efficacy was performed in TNBS-induced model of colitis by oral gavage of the NF leads, in preventive mode with a dose escalation (10, 100 and 400 mg/kg) and/or in curative mode at a single dose (100 mg/kg).
3 out of the 9 anti-TNF-alpha NFs screened at 10 mg/kg by intrarectal instillation in TNBS-induced model of colitis have demonstrated remarkable anti-inflammatory effects, with the lead NF candidate decreasing 30 % (p = 0.0008) of the lesions (5-ASA, 35 %) and 62 % of TNF-alpha expression (5-ASA, 46 %). The preventive effect of the NFs appeared to be directly correlated with their respective binding characteristics to TNF-alpha. The therapeutic efficacy of these 3 NFs in TNBS model has been improved again at the optimal dosage of 100 mg/kg, and has been further confirmed in DSS-induced model of colitis. Their anti-inflammatory efficacy was fully retained upon oral delivery without the need for a specific formulation as for the lead NF at 100 mg/kg decreasing 65 % (preventive, p < 0.00001) or 37 % (curative, p = 0.047) of intestinal inflammations.
The extreme stability of the NF scaffold allowed the generation of anti-TNF-alpha therapeutics with a powerful preventive and curative anti-inflammatory action after oral administration. The use of the oral route is expected to prevent from systemic-related side effects; making anti-TNF-alpha NFs a promising new avenue for the treatment of IBD. The drug development of the lead NFs is pursued in collaboration with a pharmaceutical partner within the SADEL FP7 European project.