Balimane Praveen V, Pace Ellen, Chong Saeho, Zhu Mingshe, Jemal Mohammed, Pelt Colleen K Van
Bristol-Myers Squibb, Princeton, NJ 08543, USA.
J Pharm Biomed Anal. 2005 Sep 1;39(1-2):8-16. doi: 10.1016/j.jpba.2005.03.043.
Parallel artificial membrane permeability assay (PAMPA) has recently gained popularity as a novel, high-throughput assay capable of rapidly screening compounds for their permeability characteristics in early drug discovery. The analytical techniques typically used for PAMPA sample analysis are HPLC-UV, LC/MS or more recently UV-plate reader. The LC techniques, though sturdy and accurate, are often labor and time intensive and are not ideal for high-throughput. On the other hand, UV-plate reader technique is amenable to high-throughput but is not sensitive enough to detect the lower concentrations that are often encountered in early drug discovery work. This article investigates a novel analytical method, a chip-based automated nanoelectrospray mass spectrometric method for its ability to rapidly analyze PAMPA permeability samples. The utility and advantages of this novel analytical method is demonstrated by comparing PAMPA permeability values obtained from nanoelectrospray to those from conventional analytical methods. Ten marketed drugs having a broad range of structural space, physico-chemical properties and extent of intestinal absorption were selected as test compounds for this investigation. PAMPA permeability and recovery experiments were conducted with model compounds followed by analysis by UV-plate reader, UV-HPLC as well as the automated nanoelectrospray technique (nanoESI-MS/MS). There was a very good correlation (r(2) > 0.9) between the results obtained using nanoelectrospray and the other analytical techniques tested. Moreover, the nanoelectrospray approach presented several advantages over the standard techniques such as higher sensitivity and ability to detect individual compounds in cassette studies, making it an attractive high-throughput analytical technique. Thus, it has been demonstrated that nanoelectrospray analysis provides a highly efficient and accurate analytical methodology to analyze PAMPA samples generated in early drug discovery.
平行人工膜渗透试验（PAMPA）作为一种新型的高通量试验，最近受到广泛关注，它能够在药物早期研发阶段快速筛选化合物的渗透特性。通常用于PAMPA样品分析的分析技术有HPLC-UV、LC/MS，或者最近使用的紫外酶标仪。LC技术虽然稳定且准确，但往往耗费人力和时间，不太适合高通量分析。另一方面，紫外酶标仪技术适合高通量分析，但灵敏度不足以检测早期药物研发工作中经常遇到的低浓度。本文研究了一种新型分析方法，即基于芯片的自动化纳米电喷雾质谱法，用于快速分析PAMPA渗透样品的能力。通过比较纳米电喷雾法与传统分析方法得到的PAMPA渗透值，证明了这种新型分析方法的实用性和优势。选择了十种具有广泛结构空间、物理化学性质和肠道吸收程度的上市药物作为本研究的测试化合物。用模型化合物进行PAMPA渗透和回收率实验，然后用紫外酶标仪、紫外HPLC以及自动化纳米电喷雾技术（nanoESI-MS/MS）进行分析。使用纳米电喷雾法与其他测试分析技术得到的结果之间具有非常好的相关性（r(2) > 0.9）。此外，纳米电喷雾方法相对于标准技术具有几个优势，如更高的灵敏度和在盒式研究中检测单个化合物的能力，使其成为一种有吸引力的高通量分析技术。因此，已经证明纳米电喷雾分析为分析早期药物研发中产生的PAMPA样品提供了一种高效、准确的分析方法。
