Anti-Human IL31RA Therapeutic Antibody (Nemolizumab)

Figure 1 Percentage reduction from baseline Owner Pruritus Visual Analog Scale (VAS) score (±SE) following subcutaneous administration of placebo or lokivetmab once on Day 0. See for number of dogs on study per treatment group at each day of study.

(A) BaF/2C6/16 cells (7.5×103/well) were cultured in 96‐well plates in RPMI‐1640 medium supplemented with 10% FBS and 100 units/mL penicillin‐100 μg/mL streptomycin and with or without hIL‐31 (0.3 ng/mL) in the presence of the indicated concentrations of nemolizumab. (B) BaF/cyIL‐31R cells (7.5×103/well) were cultured with or without cyIL‐31 (1 ng/mL) in the presence of the indicated concentrations of nemolizumab. Cell proliferation was evaluated by measuring the absorbance at 450/620 nm. (C) hIL‐31 (30 ng/mL)‐induced STAT3 phosphorylation in the presence of the various concentrations of nemolizumab in A549 cells (4×105 cells/well) was evaluated by measuring the absorbance at 450 nm as an indicator of the phosphorylation. (D‐F) Production of hIL‐31‐induced IL‐6, MMP‐1 and MMP‐3 in HaCaT cells (1×104 cells/well) in the presence of 100 ng/mL IFN‐γ was analysed by using a human IL‐6, MMP‐1 or MMP‐3 Quantikine ELISA Kit (R&D Systems). Data are expressed as the mean+SD (n=3).

Oyama, S. , Kitamura, H. , Kuramochi, T. , Higuchi, Y. , Matsushita, H. , & Suzuki, T. , et al. (2016). Cynomolgus monkey model of interleukin-31-induced scratching depicts blockade of human interleukin-31 receptor a by a humanized monoclonal antibody. Experimental Dermatology.

Figure 2 Inhibition of IL‐31‐induced scratching by nemolizumab (CIM331) in cynomolgus monkeys.

To examine the inhibitory effect of nemolizumab, the antibody was intravenously or subcutaneously administered to animals. (A) Plasma nemolizumab concentration after intravenous injection in monkey. Nemolizumab was administered to animals on Days 0 (vehicle only), 2, 7, 9, 14 and 16, as indicated by arrows. Plasma nemolizumab concentration was measured (limit of quantification: 8.00 ng/mL) on Days 3, 8, 9, 10, 14, 15, 16 and 17. Data are expressed as the mean±SEM (n=6), but bars depicting SEM are smaller than the symbols. (B) Scratching movements were counted from video recordings taken for 2 h on Days 0, 1, 3, 8, 10, 15 and 17 of monkeys that had been intravenously injected with ascending doses of nemolizumab or with vehicle (see times indicated by arrows, vehicle also given on Day 0) and then dosed with an intravenous injection of cyIL‐31 (circles) the day after. Squares denote counts from recordings taken on Days 0 (squares overlap) and 16 of monkeys that were injected with nemolizumab or vehicle but not injected with cyIL‐31. Data are expressed as mean±SEM (n=4 (vehicle), n=6 (CIM331)). (C) Nemolizumab plasma concentration‐response curve for percentage of initial scratching count on Day 1 and plasma nemolizumab concentration of nemolizumab‐treated animals at all time points. (D) Inhibition of IL‐31‐induced scratching by subcutaneously administered nemolizumab was evaluated in cynomolgus monkeys. Vehicle or 1 mg/kg of nemolizumab was administered subcutaneously to cynomolgus monkeys on Day 0. The cyIL‐31‐induced scratching was evaluated on Days −1, 28 and 56, and the basal scratching was counted on Day −2. Video‐recorded scratching behaviours were counted under a blinded manner. Data are expressed as the mean+SEM (n=6, except for the CIM331 group on Day 56 (n=5)). *P<0.05.

Oyama, S. , Kitamura, H. , Kuramochi, T. , Higuchi, Y. , Matsushita, H. , & Suzuki, T. , et al. (2016). Cynomolgus monkey model of interleukin-31-induced scratching depicts blockade of human interleukin-31 receptor a by a humanized monoclonal antibody. Experimental Dermatology.