Department of Chemical and Biomolecular Engineering, University of California Berkeley, Berkeley, CA 94720-3220, USA.
California Institute for Quantitative Biosciences (QB3), University of California Berkeley, Berkeley, CA 94720-3220, USA.
Molecules. 2023 Jul 28;28(15):5726. doi: 10.3390/molecules28155726.
Natural products provide an unparalleled diversity of small molecules to fuel drug screening efforts, but deconvoluting the pharmacological activity of natural product mixtures to identify key bioactive compounds remains a vexing and labor-intensive process. Therefore, we have developed a new platform to probe the non-specific pharmacological potential of compounds present in common dietary supplements via shotgun derivatization with isotopically labeled propanoic acid, a live cell affinity assay, which was used to selectively recognize the population of compounds which bind tightly to HeLa cells in culture, and a computational LC-MS data analysis of isotopically labeled compounds from cell lysate. The data analysis showed that hundreds of compounds were successfully derivatized in each extract, and dozens of those compounds showed high affinity for HeLa cells. In total, over a thousand isotopically labeled compounds were screened for cell affinity across three separate experiments, resulting in the identification of several known bioactive compounds with specific protein targets and six previously unreported structures. The new natural products include three tulsinol compounds which were isolated from and three valeraninium alkaloids from . The valeraninium alkaloids constitute a distinct new family of alkaloids from valerian, which may have previously undescribed bioactivity. These results collectively demonstrate the tag and snag workflow's viability as a drug discovery method.
天然产物为药物筛选工作提供了无与伦比的小分子多样性，但要从天然产物混合物中推断出其药理学活性，以确定关键的生物活性化合物，仍然是一个令人困扰且劳动密集型的过程。因此，我们开发了一种新的平台，通过用同位素标记的丙酸对常见膳食补充剂中存在的化合物进行无差别探测，进行细胞亲和力测定，该测定用于选择性识别与培养中的 HeLa 细胞紧密结合的化合物群体，以及对细胞裂解物中同位素标记化合物进行计算型 LC-MS 数据分析。数据分析表明，每种提取物中都有数百种化合物成功衍生化，其中数十种化合物对 HeLa 细胞具有高亲和力。总共在三个独立的实验中筛选了超过一千种同位素标记化合物的细胞亲和力，从而鉴定出几种具有特定蛋白质靶标的已知生物活性化合物和六种以前未报道过的结构。新的天然产物包括三种从分离的 tulsinol 化合物和三种从分离的缬草生物碱。缬草生物碱构成了缬草根中的一个独特的新生物碱家族，可能具有以前未描述的生物活性。这些结果共同证明了标记和捕捉工作流程作为一种药物发现方法的可行性。
