SSR240612 is a potent, and orally active specific non-peptide bradykinin B1 receptor antagonist (Kis = 0.48-0.73 and 358-481 nM for B1 and B2 receptors, respectively).

Bradykinin B1 (human MRC5):0.48 nM (Ki), Bradykinin B2 (human CHO-B2):358 nM (Ki), Bradykinin B1 (human HEK-B1):0.73 nM (Ki)

SSR240612阻断了[(3)H]Lys(0)-des-Arg(9)-BK与人类成纤维细胞MRC5中的B(1)受体以及在人类胚胎肾细胞中表达的重组人B(1)受体的结合，其抑制常数(K(i))分别为0.48和0.73 nM。此外，SSR240612还以1.9 nM的IC(50)抑制了Lys(0)-desAr(9)-BK (10 nM)在人类成纤维细胞MRC5中诱导的肌醇单磷酸生成。研究表明，M. tuberculosis对于所测试的拮抗剂浓度不敏感，这表明SSR240612的最小抑制浓度值大于250 μM。

SSR240612 抑制了小鼠爪子因des-Arg(9)-BK引起的水肿（口服3和10 mg/kg、腹腔注射0.3和1 mg/kg）。此外，SSR240612 减轻了小鼠耳朵由辣椒素引发的水肿（口服0.3、3和30 mg/kg），以及大鼠腹腔动脉阻塞/再灌注后肠组织破坏和中性粒细胞积累（静脉注射0.3 mg/kg）。该化合物还抑制了紫外线辐照引起的热痛觉过敏（口服1和3 mg/kg）以及大鼠对福尔马林的晚期伤害性反应（口服10和30 mg/kg）。SSR240612（口服20和30 mg/kg）预防了大鼠坐骨神经压迫引起的神经病理性热痛[1]。SSR240612 在3小时时阻断了糖饮食大鼠的触觉和寒冷痛觉过敏（ID(50)=5.5 和 7.1 mg/kg），但对照组大鼠无影响。该拮抗剂（10 mg/kg）对糖饮食大鼠的血浆葡萄糖和胰岛素、胰岛素抵抗（HOMA指数）和主动脉超氧化物阴离子产生无效[3]。

MRC5 human fibroblasts and transfected HEK-293 cells expressing human B1 receptors were routinely grown in Dulbecco's modified Eagle's medium (DMEM) with Glutamax-I supplemented with 10% fetal calf serum and antibiotics. MRC5 were incubated for 4 h in DMEM containing 0.5 μg/ml interleukin-1β (IL-1β) to induce B1 receptor synthesis. Cells were scraped and homogenized for 1 min using a Polytron (setting 8) in 25 mM TES-HCl containing 1 mM 1-10 phenantrolin. Homogenates were centrifuged at 40,000g for 15 min at 4°C, and pellets were resuspended in the same buffer using the Polytron (setting 8) for 1 min. Membranes were pelleted at 40,000g for 10 min at 4°C, resuspended in the same buffer, and conserved at 80°C. [3H]Lys0-des-Arg9-BK binding to cell membranes was performed in binding buffer of the following composition: 137 mM NaCl, 5.4 mM KCl, 1.05 mM MgCl2, 1.8 mM CaCl2, 1.2 mM NaH2PO4, 15.5 mM NaHCO3, 10 mM HEPES, 1 g/l bovine serum albumin (BSA), 140 mg/l bacitracin, and 1 μM captopril, pH 7.4. Membranes were incubated for 30 min at 25°C in 500 μl of binding buffer containing 1 nM [3H]Lys0-des-Arg9-BK for competition curves or 0.1 to 10 nM for saturation isotherms. The reaction was terminated by filtration using a Brandel Harvester onto GF/B Whatman filters previously soaked for 2 h in 0.1% polyethyleneimine. Filters were washed three times with 5 ml of binding buffer, and radioactivity was determined by liquid scintillation spectrometry. Nonspecific binding was determined by the addition of 1 μM of unlabeled Lys0 -des-Arg9 BK [1].

[3H]Inositol phosphate1 accumulation was measured in MRC5 fibroblasts labeled with [3H]myoinositol according to the method described by Oury-Donat et al. Cells cultured in 6-well plates were labeled for 48 h with 5 μCi/ml [3H]myoinositol added to the culture medium (DMEM). Cells were then incubated for 4 h in DMEM containing 0.5 μg/ml IL-1β to induce B1 receptor synthesis. Agonist stimulation of inositol phosphate 1 accumulation was performed in DMEM containing 50 mM LiCl and test compounds. Antagonists were added 15 min before 10 nM Lys0-desArg10BK. After 30 min of incubation at 37°C, the medium was discarded, and the reaction was stopped by rapid addition of 1 ml of cold methanol and 1 N HCl (v/v). The cells were scraped, and the suspension was transferred to a glass tube with 1 ml of chloroform and 20 μl of 12 N HCl. The aqueous phase was neutralized by 350 μl of 1 M NaHCO3 and applied to 1 ml of Dowex AG1 × eight columns. [3H]inositol phosphate 1 was eluted with 0.2 M ammonium formate and 0.1 M formic acid. Radioactivity was measured by liquid scintillation spectrometry [1].

Groups of eight male albino mice under isoflurane anesthesia received a 20-μl intraplantar injection into the right hind paw of 5 μg of IL-1β in phosphate-buffered saline/0.1% BSA. Forty minutes later (T = 0), mice received, under anesthesia, a 20-μl intraplantar injection in the same paw of des-Arg9-BK (10 μg/paw) in water. SSR240612 or vehicle [5% (v/v) ethanol and 5% (v/v) Tween 80 in water] was administered by oral route at the doses of 1, 3, and 10 mg/kg 1 h before des-Arg9-BK injection and by intraperitoneal route at the doses of 0.1, 0.3, and 1 mg/kg 40 min before des-Arg9-BK injection. Paw volume was measured with a plethysmometer at T =-2 h (initial measurement) and at several times after edema induction (T = 20, 40, 60, and 120 min). Paw edema volume was expressed in milliliters as the difference between the paw volume at each time after edema induction and the initial paw volume. Results for each group are expressed as mean ± S.E.M. of individual paw edema volumes [1].