Kilcoyne Jane, Twiner Michael J, McCarron Pearse, Crain Sheila, Giddings Sabrina D, Foley Barry, Rise Frode, Hess Philipp, Wilkins Alistair L, Miles Christopher O
†Marine Institute, Rinville, Oranmore, County Galway, Ireland.
‡School of Chemical and Pharmaceutical Sciences, Dublin Institute of Technology, Kevin Street, Dublin 8, Ireland.
J Agric Food Chem. 2015 May 27;63(20):5083-91. doi: 10.1021/acs.jafc.5b01320. Epub 2015 May 12.
Azaspiracids (AZAs) are marine biotoxins produced by dinoflagellates that can accumulate in shellfish, which if consumed can lead to poisoning events. AZA7-10, 7-10, were isolated from shellfish and their structures, previously proposed on the basis of only LC-MS/MS data, were confirmed by NMR spectroscopy. Purified AZA4-6, 4-6, and 7-10 were accurately quantitated by qNMR and used to assay cytotoxicity with Jurkat T lymphocyte cells for the first time. LC-MS(MS) molar response studies performed using isocratic and gradient elution in both selected ion monitoring and selected reaction monitoring modes showed that responses for the analogues ranged from 0.3 to 1.2 relative to AZA1, 1. All AZA analogues tested were cytotoxic to Jurkat T lymphocyte cells in a time- and concentration-dependent manner; however, there were distinct differences in their EC50 values, with the potencies for each analogue being: AZA6 > AZA8 > AZA1 > AZA4 ≈ AZA9 > AZA5 ≈ AZA10. This data contributes to the understanding of the structure-activity relationships of AZAs.
azaspiracids（AZAs）是由鞭毛藻产生的海洋生物毒素，可在贝类中蓄积，食用后可能导致中毒事件。从贝类中分离出了AZA7 - 10，其结构先前仅基于LC - MS/MS数据提出，现通过核磁共振光谱得以确认。通过定量核磁共振对纯化的AZA4 - 6和7 - 10进行了准确的定量，并首次用于检测对Jurkat T淋巴细胞的细胞毒性。在选择离子监测和选择反应监测模式下，使用等度洗脱和梯度洗脱进行的LC - MS（MS）摩尔响应研究表明，相对于AZA1，各类似物的响应范围为0.3至1.2。所有测试的AZA类似物均对Jurkat T淋巴细胞具有时间和浓度依赖性的细胞毒性；然而，它们的半数有效浓度（EC50）值存在明显差异，各类似物的效力顺序为：AZA6 > AZA8 > AZA1 > AZA4 ≈ AZA9 > AZA5 ≈ AZA10。该数据有助于理解AZAs的构效关系。
