| 名稱 | SSR240612 |
| 描述 | 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). |
| 細(xì)胞實(shí)驗(yàn) | [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]. |
| 激酶實(shí)驗(yàn) | 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]. |
| 動物實(shí)驗(yàn) | 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]. |
| 體外活性 | SSR240612阻斷了[(3)H]Lys(0)-des-Arg(9)-BK與人類成纖維細(xì)胞MRC5中的B(1)受體以及在人類胚胎腎細(xì)胞中表達(dá)的重組人B(1)受體的結(jié)合，其抑制常數(shù)(K(i))分別為0.48和0.73 nM。此外，SSR240612還以1.9 nM的IC(50)抑制了Lys(0)-desAr(9)-BK (10 nM)在人類成纖維細(xì)胞MRC5中誘導(dǎo)的肌醇單磷酸生成。研究表明，M. tuberculosis對于所測試的拮抗劑濃度不敏感，這表明SSR240612的最小抑制濃度值大于250 μM。 |
| 體內(nèi)活性 | SSR240612 抑制了小鼠爪子因des-Arg(9)-BK引起的水腫（口服3和10 mg/kg、腹腔注射0.3和1 mg/kg）。此外，SSR240612 減輕了小鼠耳朵由辣椒素引發(fā)的水腫（口服0.3、3和30 mg/kg），以及大鼠腹腔動脈阻塞/再灌注后腸組織破壞和中性粒細(xì)胞積累（靜脈注射0.3 mg/kg）。該化合物還抑制了紫外線輻照引起的熱痛覺過敏（口服1和3 mg/kg）以及大鼠對福爾馬林的晚期傷害性反應(yīng)（口服10和30 mg/kg）。SSR240612（口服20和30 mg/kg）預(yù)防了大鼠坐骨神經(jīng)壓迫引起的神經(jīng)病理性熱痛[1]。SSR240612 在3小時(shí)時(shí)阻斷了糖飲食大鼠的觸覺和寒冷痛覺過敏（ID(50)=5.5 和 7.1 mg/kg），但對照組大鼠無影響。該拮抗劑（10 mg/kg）對糖飲食大鼠的血漿葡萄糖和胰島素、胰島素抵抗（HOMA指數(shù)）和主動脈超氧化物陰離子產(chǎn)生無效[3]。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | H2O : Insoluble
DMSO : 100 mg/mL (126 mM)
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| 關(guān)鍵字 | SSR 240612 | SSR240612 | Inhibitor | inhibit | SSR-240612 | Bradykinin Receptor |
| 相關(guān)產(chǎn)品 | B-Raf IN 14 | [Des-Arg9]-Bradykinin acetate | Sar-[D-Phe8]-des-Arg9-Bradykinin acetate | Lys-Bradykinin acetate(342-10-9 free base) | MK-0686 | Bradykinin (2-9) | 5-Hydroxy-1-methylhydantoin | NPC 567 acetate | Safotibant | Anatibant 2HCl | (Hyp3)-Bradykinin acetate | Lys-[Des-Arg9]Bradykinin acetate |
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