Aclidinium bromide (LAS-W 330) is a synthetic anticholinergic agent that is used as an inhalant for treatment of acute bronchospasm due to chronic bronchitis or emphysema. Aclidinium has not been implicated in causing liver enzyme elevations or clinically apparent acute liver injury.

M5 mAChR:0.16 nM(Ki), M1 mAChR:0.1 nM(Ki), M3 mAChR:0.14 nM(Ki), M2 mAChR:0.14 nM(Ki), M4 mAChR:0.21 nM(Ki)

Aclidinium在所研究的所有物种的血浆样品中均被水解,37℃下,在大鼠,豚鼠,狗和人的血浆中,表观半衰期分别为11.7分钟,38.3分钟,1.8分钟和2.4分钟。0.1 μM Aclidinium抑制人支气管成纤维细胞中乙酰胆碱和TGF-β1诱导的Ⅰ型胶原上调,并抑制α-SMA mRNA和蛋白质表达。0.1 μM Aclidinium抑制碳酰胆碱和TGF-β1诱导的人支气管成纤维细胞中ERK1/2磷酸化和RhoA-GTP形成的增加。0.1 μM Aclidinium抑制TGF-β1诱导的人支气管成纤维细胞中ChAT表达的上调。0.1 μM Aclidinium在剂量依赖性地抑制人肺成纤维细胞的TGF-β1和卡巴胆碱诱导的细胞增殖。< 100 nM Aclidinium剂量依赖性抑制豚鼠离体豚鼠气管中卡巴胆碱诱导的收缩。Aclidinium预处理可阻止人肺成纤维细胞中M1和M3的上调,但不能抑制由碳酰胆碱或TGF-β1诱导的M2的下调。

500 μg/kg Aclidinium给药1小时后诱导有意识的比格犬心率最大增加55%.1 mg/mL Aclidinium在超过120分钟的研究期间,产生对麻醉的豚鼠有效持久的气管保护作用(72%–88.4%).在麻醉的豚鼠的乙酰胆碱诱导的支气管收缩模型中,[3H] AcliAclidinium显示作用开始,IC50（95％CI）为140 μg/ mL,tmax为30分钟.

Affinity assay: The affinity of Aclidinium for the different human muscarinic receptor subtypes at equilibrium is determined by measuring their ability to displace the binding of [3H]NMS to cell membrane preparations expressing one of the human muscarinic receptor subtypes. Protein concentrations are 8.1 μg/well, 10.0 μg/well, 4.9 μg/well, 4.5 μg/well, and 5.0 μg/well for M1, M2, M3, M4, and M5 receptor membrane preparations, respectively. The assays are conducted at [3H]NMS concentrations approximately equal to the radioligand equilibrium dissociation constant (Kd) for the different muscarinic receptors subtypes. The [3H]NMS concentration is 0.3 nM for the M1 and M4 assays and 1 nM for the M2, M3, and M5 assays. A range of antagonist concentrations (10?14 to 10?5 M) are tested in duplicate to generate competition curves. Nonspecific binding is determined in the presence of atropine (1 μM). Assay reagents are dissolved in assay binding buffer (phosphate-buffered saline with calcium and magnesium) to a total volume of 200 μL. After a 2 hours or 6 hours incubation period (M1–M4 and M5, respectively) at room temperature in 96-well microtiter plates to ensure that equilibrium is achieved for Aclidinium, 150 μL aliquots of the reaction are transferred to GF/C filter plates pretreated for 1 hour with wash buffer (50 mM Tris, 100 mM NaCl, pH 7.4) containing 0.05% polyethylenimine. Bound and free [3H]NMS are then separated by rapid vacuum filtration followed by four washes with ice-cold wash buffer. Filters are then dried for 30 min before addition of 30 μL of OptiPhase Supermix, and radioactivity is quantified using a MicroBeta Trilux microplate scintillation counter.

Human bronchial fibroblast proliferation is measured as previously outlined by colorimetric immunoassay based on BrdU incorporation during DNA synthesis using a cell proliferation enzyme-linked immunosorbent assay BrdU kit according to the manufacturer's protocol. Cells are seeded at a density of 3x103 cells/well on 96-well plates and incubated for 24 hours. Cells are then exposed to different experimental conditions. The 490 nm absorbance is quantified using a microplate spectrophotometer. Proliferation data refers to the absorbance values of BrdU-labeled cellular DNA content per well. Stimulation is expressed as x-fold proliferation over basal growth of the untreated control set as unity.(Only for Reference)