PMID- 26352810 OWN - NLM STAT- MEDLINE DCOM- 20160524 LR - 20181113 IS - 1932-6203 (Electronic) IS - 1932-6203 (Linking) VI - 10 IP - 9 DP - 2015 TI - Pharmacokinetic and Pharmacodynamic Characterisation of an Anti-Mouse TNF Receptor 1 Domain Antibody Formatted for In Vivo Half-Life Extension. PG - e0137065 LID - 10.1371/journal.pone.0137065 [doi] LID - e0137065 AB - Tumour Necrosis Factor-alpha (TNF-alpha) inhibition has been transformational in the treatment of patients with inflammatory disease, e.g. rheumatoid arthritis. Intriguingly, TNF-alpha signals through two receptors, TNFR1 and TNFR2, which have been associated with detrimental inflammatory and beneficial immune-regulatory processes, respectively. To investigate if selective TNFR1 inhibition might provide benefits over pan TNF-alpha inhibition, tools to investigate the potential impact of pharmacological intervention are needed. Receptor-deficient mice have been very insightful, but are not reversible and could distort receptor cross-talk, while inhibitory anti-TNFR1 monoclonal antibodies have a propensity to induce receptor agonism. Therefore, we set out to characterise a monovalent anti-TNFR1 domain antibody (dAb) formatted for in vivo use. The mouse TNFR1 antagonist (DMS5540) is a genetic fusion product of an anti-TNFR1 dAb with an albumin-binding dAb (AlbudAb). It bound mouse TNFR1, but not human TNFR1, and was an antagonist of TNF-alpha-mediated cytotoxicity in a L929 cell assay. Surprisingly, the dAb did not compete with TNF-alpha for TNFR1-binding. This was supported by additional data showing the anti-TNFR1 epitope mapped to a single residue in the first domain of TNFR1. Pharmacokinetic studies of DMS5540 in mice over three doses (0.1, 1.0 and 10 mg/kg) confirmed extended in vivo half-life, mediated by the AlbudAb, and demonstrated non-linear clearance of DMS5540. Target engagement was further confirmed by dose-dependent increases in total soluble TNFR1 levels. Functional in vivo activity was demonstrated in a mouse challenge study, where DMS5540 provided dose-dependent inhibition of serum IL-6 increases in response to bolus mouse TNF-alpha injections. Hence, DMS5540 is a potent mouse TNFR1 antagonist with in vivo pharmacokinetic and pharmacodynamic properties compatible with use in pre-clinical disease models and could provide a useful tool to dissect the individual contributions of TNFR1 and TNFR2 in homeostasis and disease. FAU - Goodall, Laura J AU - Goodall LJ AD - Biopharm Innovation Unit, Biopharm R&D, GlaxoSmithKline, Stevenage, United Kingdom. FAU - Ovecka, Milan AU - Ovecka M AD - Biopharm Innovation Unit, Biopharm R&D, GlaxoSmithKline, Stevenage, United Kingdom. FAU - Rycroft, Daniel AU - Rycroft D AD - Biopharm Innovation Unit, Biopharm R&D, GlaxoSmithKline, Stevenage, United Kingdom. FAU - Friel, Sarah L AU - Friel SL AD - Biopharm Innovation Unit, Biopharm R&D, GlaxoSmithKline, Stevenage, United Kingdom. FAU - Sanderson, Andrew AU - Sanderson A AD - Biopharm Innovation Unit, Biopharm R&D, GlaxoSmithKline, Stevenage, United Kingdom. FAU - Mistry, Prafull AU - Mistry P AD - R&D Projects, Clinical Platforms and Sciences, GlaxoSmithKline, Stevenage, United Kingdom. FAU - Davies, Marie L AU - Davies ML AD - Biopharm Innovation Unit, Biopharm R&D, GlaxoSmithKline, Stevenage, United Kingdom. FAU - Stoop, A Allart AU - Stoop AA AD - Biopharm Innovation Unit, Biopharm R&D, GlaxoSmithKline, Stevenage, United Kingdom. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150909 PL - United States TA - PLoS One JT - PloS one JID - 101285081 RN - 0 (Antibodies, Monoclonal) RN - 0 (DMS5540) RN - 0 (Epitopes) RN - 0 (Interleukin-6) RN - 0 (Receptors, Tumor Necrosis Factor, Type I) RN - 0 (Recombinant Fusion Proteins) RN - 0 (Single-Domain Antibodies) RN - 0 (Tnfrsf1a protein, mouse) RN - 0 (Tumor Necrosis Factor-alpha) SB - IM MH - Animals MH - Antibodies, Monoclonal/*administration & dosage/genetics/pharmacokinetics MH - Arthritis, Rheumatoid/blood/immunology/*therapy MH - Cell Line MH - Epitopes/drug effects/immunology MH - Humans MH - Interleukin-6/blood MH - Mice MH - Receptors, Tumor Necrosis Factor, Type I/antagonists & inhibitors/genetics/*immunology MH - Recombinant Fusion Proteins/*administration & dosage MH - Signal Transduction MH - Single-Domain Antibodies/*administration & dosage MH - Tumor Necrosis Factor-alpha/antagonists & inhibitors/immunology/*metabolism PMC - PMC4564187 COIS- Competing Interests: The authors have read the journal's policy and have the following conflicts: All authors were employees of GlaxoSmithKline at the time this work was carried out and received GSK shares as part of their remuneration. AS and AAS are named inventors on patents assigned to GlaxoSmithKline relating to TNFR1 antagonist domain antibodies (WO2010/094720, WO2011/051217 and WO2012/172070). All authors were employees of GlaxoSmithKline at the time this work was carried out. All relevant patents have been declared, and there are no products in development or marketed products to declare. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors. EDAT- 2015/09/10 06:00 MHDA- 2016/05/25 06:00 PMCR- 2015/09/09 CRDT- 2015/09/10 06:00 PHST- 2014/12/20 00:00 [received] PHST- 2015/08/12 00:00 [accepted] PHST- 2015/09/10 06:00 [entrez] PHST- 2015/09/10 06:00 [pubmed] PHST- 2016/05/25 06:00 [medline] PHST- 2015/09/09 00:00 [pmc-release] AID - PONE-D-14-55631 [pii] AID - 10.1371/journal.pone.0137065 [doi] PST - epublish SO - PLoS One. 2015 Sep 9;10(9):e0137065. doi: 10.1371/journal.pone.0137065. eCollection 2015.