Sun Hai-Yan, Guan Su, Bi Hui-Chang, Su Qi-Biao, Huang Wen-Lin, Chowbay Balram, Huang Min, Chen Xiao, Li Chun-Guang, Zhou Shu-Feng
Institute of Clinical Pharmacology, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510080, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Jul 1;854(1-2):320-7. doi: 10.1016/j.jchromb.2007.03.052. Epub 2007 Apr 19.
4-Anilinoquinazolines (e.g. Iressa and Glivec) are a class of epidermal growth factor receptor tyrosine kinase (EGFR-TK) inhibitors widely used to treat non-small cell lung cancer and other tumors. However, low clinical response rate, resistance, and host toxicity of currently available EGFR-TK inhibitors prompt the development of second generation of TK inhibitors with improved efficacy, selectivity, and less resistance. CH330331 is a recently synthesized novel 4-anilinoquinazoline analog with confirmed anticancer activity in vitro and in vivo. To predict its oral pharmacokinetic behavior and transport nature in the intestine before entering clinical trials, we have developed and validated a high performance liquid chromatographic (HPLC) method for the determination of CH330331 in Caco-2 (a human colon cancer cell line) monolayers. The developed HPLC method was sensitive and reliable, with acceptable accuracy (90-110% of nominal values) and precision (intra- and inter-assay R.S.D.<10%). The total running time was within 10 min, with acceptable separation of the target analytes. The lower limit of quantitation (LLOQ) value for CH330331 was 200 ng/ml when an aliquot of 100 microl sample was injected onto the HPLC. The validated HPLC method was applied to characterize the epithelial transport of CH330331 in Caco-2 monolayers. The transport of CH330331 across the Caco-2 monolayers from the apical to basolateral side was 8- to 10-fold higher than that from the basolateral to apical side. Co-incubation of sodium azide or MK-571, but not verapamil, significantly inhibited the apical to basolateral transport of CH330331. These findings provide initial evidence that the intestinal absorption of CH330331 is mediated by an active mechanism. Further studies are required to explore the interaction of CH330331 with ATP-binding cassette transporters and the possible influence on its pharmacokinetics and pharmacodynamics.
4-苯胺基喹唑啉类药物（如易瑞沙和格列卫）是一类表皮生长因子受体酪氨酸激酶（EGFR-TK）抑制剂，广泛用于治疗非小细胞肺癌和其他肿瘤。然而，目前可用的EGFR-TK抑制剂临床反应率低、存在耐药性以及宿主毒性，促使人们开发具有更高疗效、选择性和更低耐药性的第二代TK抑制剂。CH330331是最近合成的一种新型4-苯胺基喹唑啉类似物，已证实其在体外和体内均具有抗癌活性。为了在进入临床试验之前预测其口服药代动力学行为以及在肠道中的转运特性，我们开发并验证了一种高效液相色谱（HPLC）方法，用于测定Caco-2（一种人结肠癌细胞系）单层细胞中CH330331的含量。所开发得HPLC方法灵敏且可靠，准确度（为标称值的90 - 110%）和精密度（批内和批间相对标准偏差<10%）均可接受。总运行时间在10分钟以内，目标分析物分离效果良好。当将100微升样品注入HPLC时，CH330331的定量下限（LLOQ）值为200纳克/毫升。经验证的HPLC方法用于表征CH330331在Caco-2单层细胞中的上皮转运。CH330331从顶侧到基底外侧穿过Caco-2单层细胞层的转运比从基底外侧到顶侧的转运高8至10倍。叠氮化钠或MK - 571共同孵育可显著抑制CH330331从顶侧到基底外侧的转运，但维拉帕米则无此作用。这些发现提供了初步证据，表明CH330331的肠道吸收是由一种主动机制介导的。需要进一步研究来探索CH330331与ATP结合盒转运体的相互作用及其对药代动力学和药效学的可能影响。
