Ab501

Adult male Balb/c mice (aged 6–8 weeks, vehicle n = 8; UCB-9260n = 8 and AB501 n = 6 mice, from Charles River UK) received 4 mg of anti-collagen type II antibody cocktail intraperitoneally, which was followed 24 h later with 25 µg of lipopolysaccharide (LPS) intraperitoneally. Dosing with vehicle (30% cyclodextrin po bid) or UCB-9260 (150 mg/kg po bid) was initiated 6 h post-LPS administration. AB501 (in-house anti-mouse TNF antibody, 100 m/kg) was administered intravenously 6 h post-LPS administration followed by three further doses administered subcutaneously at 48-h intervals. Animals were scored for signs of arthritis and weighed daily. The following scoring system was used for signs of arthritis: 0-no arthritis, 1-wrist/ankle affected, 2-wrist/ankle and pad affected and 3-wrist/ankle, pad, and digits affected. Data shown as the mean ± standard error of arthritis clinical score for the treatment group. Statistical significance (*p < 0.05, **p < 0.01) was determined using one-way ANOVA with Dunnett’s multiple comparison test.
All in vivo studies were reviewed by an internal ethical review body and conducted in accordance with the Animals (Scientific Procedures) Act 1986, EU Directive 2010/63/EU. Animals were kept under a light/dark cycle of 12/12 h and had access to food and water ad libitum.

Adult male Balb/c mice (aged 6–8 weeks, PBS n = 5; vehicle n = 8; UCB-9260 10, 30, 100 and 300 mg/kg n = 8; Ab501 n = 3 mice, from Charles River UK) were dosed intravenously with 30 mg/kg of Ab501 (in-house anti-mouse TNF antibody) diluted in PBS 1 h prior to zymosan challenge. Vehicle (30% cyclodextrin) or UCB-9260 at 10, 30, 100 and 300 mg/kg was dosed orally at the time of zymosan challenge. zymosan (R&D Systems) was administered by intraperitoneal injection at a concentration of 1 µg per mouse in PBS (100 µL). One hour post oral administration of UCB-9260, mice were bled via the tail to analyse exposure levels of the compound. Four hours after the zymosan challenge, mice were sacrificed and the peritoneal cavity lavaged with 3 mL of FACS buffer. An aliquot (0.3 mL) of lavage fluid was used to assess the number of CD45⁺GR1⁺ cells by flow cytometry. Briefly, the cells were treated with ACK lysis buffer, washed and suspended in FACS buffer prior to staining with FITC anti-mouse CD45 and PE anti-mouse GR1 (BD Biosciences). Data are shown as mean ± standard error of the number of CD45⁺GR-1⁺ cells. Statistical significance (*p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001) was determined using one-way ANOVA with Dunnett’s multiple comparison test.