Brent Kern1, Wenlin Li1, Christine Bono2, Li-Fen Lee3, Eugenia Kraynov1. 1. Pharmacokinetics, Dynamics, and Metabolism, Pfizer, La Jolla, California. 2. Drug Safety Research and Development, Pfizer, Groton, Connecticut. 3. Rinat, Experimental Medicine, Pfizer, San Francisco, California.
Abstract
BACKGROUND: Interleukin-7 receptor α (IL-7Rα) is associated with autoimmune disease. Blocking its activation by interleukin-7 (IL-7) with a therapeutic monoclonal antibody may reduce pathogenic T cells and effectively control the autoimmune response in these disorders. METHODS: Two flow cytometry-based assays were developed and implemented to evaluate the interaction between cell surface IL-7Rα and an anti-IL-7Rα monoclonal antibody (Ab1). The receptor occupancy assay utilized competing and noncompeting commercial detection antibodies for "free" and "total" IL-7Rα, respectively. STAT5 phosphorylation (pSTAT5) was measured as a proximal biomarker of IL-7Rα inhibition by Ab1. RESULTS: Monkeys administered Ab1 had no free IL-7Rα detectable on the CD3+ T cell surface at 0.25 hours postdose through day 4, in all treatment groups. Ab1 treatment resulted in a significant reduction in total IL-7Rα, dropping to 53%, 44%, and 55% on day 4 at 0.3, 3, and 30 mg/kg, respectively, compared to predose levels. There were treatment-related decreases in the ability of IL-7 to induce STAT5 phosphorylation in both CD4+ and CD8+ T cells in monkey blood samples from all treated animals from 0.25 hours through Day 4 postdose. CONCLUSIONS: The nonclinical receptor occupancy assay was developed and applied to detect free and total IL-7Rα on the surface of CD3+ T cells in cynomolgus monkeys treated with Ab1. The results showed good correlation with the phosphorylation of STAT5 and serum concentration of Ab1. The approach for IL-7Rα occupancy and pSTAT5 measurements established in monkeys can be utilized in clinical trials for pharmacokinetic/pharmacodynamic evaluation of Ab1 effect in humans.
BACKGROUND:Interleukin-7 receptor α (IL-7Rα) is associated with autoimmune disease. Blocking its activation by interleukin-7 (IL-7) with a therapeutic monoclonal antibody may reduce pathogenic T cells and effectively control the autoimmune response in these disorders. METHODS: Two flow cytometry-based assays were developed and implemented to evaluate the interaction between cell surface IL-7Rα and an anti-IL-7Rα monoclonal antibody (Ab1). The receptor occupancy assay utilized competing and noncompeting commercial detection antibodies for "free" and "total" IL-7Rα, respectively. STAT5 phosphorylation (pSTAT5) was measured as a proximal biomarker of IL-7Rα inhibition by Ab1. RESULTS: Monkeys administered Ab1 had no free IL-7Rα detectable on the CD3+ T cell surface at 0.25 hours postdose through day 4, in all treatment groups. Ab1 treatment resulted in a significant reduction in total IL-7Rα, dropping to 53%, 44%, and 55% on day 4 at 0.3, 3, and 30 mg/kg, respectively, compared to predose levels. There were treatment-related decreases in the ability of IL-7 to induce STAT5 phosphorylation in both CD4+ and CD8+ T cells in monkey blood samples from all treated animals from 0.25 hours through Day 4 postdose. CONCLUSIONS: The nonclinical receptor occupancy assay was developed and applied to detect free and total IL-7Rα on the surface of CD3+ T cells in cynomolgus monkeys treated with Ab1. The results showed good correlation with the phosphorylation of STAT5 and serum concentration of Ab1. The approach for IL-7Rα occupancy and pSTAT5 measurements established in monkeys can be utilized in clinical trials for pharmacokinetic/pharmacodynamic evaluation of Ab1 effect in humans.
Authors: Lyssia Belarif; Caroline Mary; Lola Jacquemont; Hoa Le Mai; Richard Danger; Jeremy Hervouet; David Minault; Virginie Thepenier; Veronique Nerrière-Daguin; Elisabeth Nguyen; Sabrina Pengam; Eric Largy; Arnaud Delobel; Bernard Martinet; Stéphanie Le Bas-Bernardet; Sophie Brouard; Jean-Paul Soulillou; Nicolas Degauque; Gilles Blancho; Bernard Vanhove; Nicolas Poirier Journal: Nat Commun Date: 2018-10-26 Impact factor: 14.919
Authors: Joanne Ellis; Andre van Maurik; Lea Fortunato; Sophie Gisbert; Keguan Chen; Ann Schwartz; Simon McHugh; Andrew Want; Sara Santos Franco; Joao-Joaquim Oliveira; Jeffrey Price; Alasdair Coles; Kim Brown; Donggang Su; Jenny L Craigen; Jiansong Yang; Sara Brett; Bill Davis; Joseph Cheriyan; Onajite Kousin-Ezewu; Frank Gray; Paul W Thompson; Disala Fernando Journal: Br J Clin Pharmacol Date: 2018-12-03 Impact factor: 4.335