Pellowska M, Stein C, Pohland M, Merk D, Klein J, Eckert G P, Schubert-Zsilavecz M, Wurglics M
Institute of Pharmaceutical Chemistry, Goethe-University, Max-von-Laue Str. 9, D-60438 Frankfurt, Germany.
Department of Pharmacology, Goethe-University, Max-von-Laue Str. 9, D-60438 Frankfurt, Germany.
J Pharm Biomed Anal. 2015 Jan;102:417-24. doi: 10.1016/j.jpba.2014.10.001. Epub 2014 Oct 13.
Alzheimer's disease (AD) is the most common cause of dementia. Since no causative treatment is available, new therapeutic options are utmost needed. Several pirinixic acid derivatives, including MH84 (2-((4,6-bis(4-(trifluoromethyl)phenethoxy)pyrimidin-2-yl)thio)hexanoic acid), have shown promising in vitro results as γ-secretase modulators as well as PPARγ activators as potential pharmacological compounds against AD. Using a newly developed and validated sensitive LC-MS (APCI-qTOF mass analyzer) method, the pharmacokinetic and long-term accumulating properties as well as the blood-brain-barrier permeability of MH84 were evaluated in a preclinical animal study. MH84 was administered to mice by oral gavage with a dose of 12 mg/kg. Nine time points from 0.5 to 48 h with 6 animals per point were investigated. Additionally 6 animals were fed daily, for 21 days with an identical dose to determine possible long-term accumulation in plasma and brain tissue. The sample preparation was performed by a liquid-liquid extraction on Extrelut(®) columns whereas the LC separation was operated on a MulthoHigh 100 RP 18-5 μ column (125 × 4 mm) using an isocratic mobile phase of formic acid (0.1% (v/v))-methanol mixture (11:89 (v/v)) at a flow rate of 1 ml/min. The validation confirmed the new LC-MS method to be precise, accurate and reliable. After oral application, Cmax and Tmax of unmetabolized MH84 was determined to be 10.90 μg/ml and 3h in plasma. In brain tissue a constant level of 300 to maximum 320.64 ng/g was found after 1.5-6h. Daily gavage for 21 days did not lead to a long-term drug accumulation in the brain. The efficacy of the obtained MH84 levels needs to be investigated in further preclinical pharmacodynamic animal studies.
阿尔茨海默病（AD）是痴呆最常见的病因。由于尚无病因性治疗方法，因此极其需要新的治疗选择。包括MH84（2-((4,6-双(4-(三氟甲基)苯乙氧基)嘧啶-2-基)硫代)己酸）在内的几种匹立尼酸衍生物，作为γ-分泌酶调节剂以及PPARγ激活剂，在体外作为抗AD的潜在药理化合物已显示出有前景的结果。在一项临床前动物研究中，使用新开发并经验证的灵敏液相色谱-质谱联用（大气压化学电离-四极杆飞行时间质谱仪）方法，评估了MH84的药代动力学和长期蓄积特性以及血脑屏障通透性。通过口服灌胃以12 mg/kg的剂量给小鼠施用MH84。研究了从0.5至48小时的9个时间点，每个时间点6只动物。另外，6只动物每天以相同剂量喂食21天，以确定在血浆和脑组织中可能的长期蓄积情况。样品制备通过在Extrelut®柱上进行液-液萃取来完成，而液相色谱分离则在MulthoHigh 100 RP 18 - 5μm柱（125×4 mm）上进行，使用甲酸（0.1%（v/v））-甲醇混合物（11:89（v/v））的等度流动相，流速为1 ml/min。验证证实新的液相色谱-质谱联用方法精确、准确且可靠。口服给药后，血浆中未代谢的MH84的Cmax和Tmax分别确定为10.90μg/ml和3小时。在脑组织中，1.5 - 6小时后发现恒定水平为300至最高320.64 ng/g。每天灌胃21天未导致药物在脑中的长期蓄积。所获得的MH84水平的疗效需要在进一步的临床前药效学动物研究中进行研究。
