Ramos Ana Cláudia M, Rezende Kênnia R, Teixeira Carolina M, Fernandes Aline R, Santos Heloa, Machado Rúbia Darc, Menegatti Ricardo, Vaz Boniek G, Chaves Andréa R
Laboratory of Biopharmacy and Pharmacokinetics (BioPK), Faculty of Pharmacy, Federal University of Goiás, Goiânia 74605-170, GO, Brazil.
Laboratory of Chromatography and Mass Spectrometry (LaCEM), Chemistry Institute, Federal University of Goiás, Goiânia 74690-900, GO, Brazil.
Pharmaceuticals (Basel). 2023 Jun 26;16(7):930. doi: 10.3390/ph16070930.
The LQFM05 is a prototype drug designed for treatment of psychiatric disorders, such as schizophrenia, exhibiting anxiolytic- and antidepressant-like (12 or 24 µmol/kg) effects in classical behavioral tests. In order to evaluate its pharmacokinetic properties, a liquid chromatography method coupled to a quadrupole time of flight mass spectrometry system (LC-QTOF/MS) was developed and fully validated for LQFM05 analysis in rat plasma and tissue samples (brain, heart, liver, and kidneys). Liquid-liquid extraction, solid phase extraction and protein precipitation were assessed as clean-up procedures for biological samples and analyte enrichment. Plasma and tissue samples underwent protein precipitation as a preliminary step, using acetonitrile. Linearity was fully demonstrated for the dynamic range (10.0 to 900.0 ng/mL), with r values higher than 0.99 (RSD ≤ 2%, F < F, C < C). Biodistribution studies in rats revealed high brain tissue concentrations (12.4 µg/g), suggesting elevated drug affinity to the main therapeutic target tissue, showing a blood partition coefficient of 1.9. Kidneys also showed great exposure and tissue affinity, suggesting a potential extrahepatic clearance. Likewise, all examined tissues exhibited satisfactory LQFMF05 distribution. The mass fragmentation spectrum indicated the presence of its main metabolite, LQFM235, yielded by high hepatic hydroxylation route, an equally bioactive derivative. Lastly, the developed LC-QTOF/MS method was shown to be sensitive (LOQ = 10 ng/mL), precise and accurate for LQFM05 determination in tissue homogenates and plasma samples.
LQFM05是一种设计用于治疗精神疾病（如精神分裂症）的原型药物，在经典行为测试中表现出抗焦虑和抗抑郁样（12或24 μmol/kg）效应。为了评估其药代动力学性质，开发了一种液相色谱法与四极杆飞行时间质谱系统（LC-QTOF/MS）联用的方法，并对大鼠血浆和组织样本（脑、心脏、肝脏和肾脏）中的LQFM05分析进行了全面验证。液液萃取、固相萃取和蛋白沉淀作为生物样本净化程序和分析物富集方法进行了评估。血浆和组织样本首先使用乙腈进行蛋白沉淀。在动态范围（10.0至900.0 ng/mL）内充分证明了线性，r值高于0.99（RSD≤2%，F<F，C<C）。大鼠体内生物分布研究显示脑组织浓度较高（12.4 μg/g），表明药物对主要治疗靶组织的亲和力较高，血液分配系数为1.9。肾脏也显示出较高的暴露量和组织亲和力，表明可能存在肝外清除。同样，所有检测组织均表现出令人满意的LQFMF（此处原文有误，应为LQFM05）分布。质谱碎片谱表明存在其主要代谢产物LQFM235，它是通过高肝羟基化途径产生的一种同样具有生物活性的衍生物。最后，所开发的LC-QTOF/MS方法对组织匀浆和血浆样本中LQFM05的测定显示出灵敏（定量限=10 ng/mL）、精确且准确。
