Diacylglycerol Lactones as C1 Ligands for Protein Kinase C and Related Proteins

继续研究新的二酰基甘油-内酯，目的是针对单个蛋白激酶C同工酶和其他含有类似C1结构域的二酰基甘油反应蛋白。选择分子靶标研究所JDC审议的两个DAG-内酯是在更大范围内合成的，并在NCI 60细胞系面板上进行测试。这些化合物仍然是进一步考虑的有力候选者，等待NCI成像和分子治疗科主任Kevin A.Camphausen博士的进一步研究结果。合成并研究了由刚性4-[(甲基苯基)乙炔]苯基与DAG-内酯核的外环酰基羰基通过可变长度的间隔基团隔开的二酰基甘油内酯。测定了一组经典的和新型的PKC同工酶在两种不同的磷脂环境中的结合亲和力，其中一个对应于细胞的质膜。研究了这两种化合物处理人前列腺癌LNCaP细胞后，PKC同工酶的动力学和易位位置，以及ERK磷酸化的早期反应和诱导细胞凋亡的晚期反应。研究发现，DAG-内酯上刚性结构基序的不同空间配置导致了活性的差异。

New diacylglycerol-lactones continued to be studied with the aim of targeting individual protein kinase C isozymes and other diacylglycerol-responsive proteins containing similar C1 domains. The two DAG-lactones selected for consideration by the JDC of the Molecular Targets Faculty were synthesized in larger scale and tested against the NCI 60-cell line panel. The compounds are still strong candidates for further consideration pending results from additional studies by Dr. Kevin A. Camphausen, Head, Imaging and Molecular Therapeutics Section, NCI. Additional diacylglycerol lactones built with a rigid 4-[(methylphenyl)ethynyl]phenyl rod separated from the exocyclic acylcarbonyl of the DAG-lactone core by a spacer unit of variable length were synthesized and studied. Binding affinities for a panel of classical and novel PKC isozymes in two different phospholipids environments, one corresponding to the plasma membrane of cells, were determined. The kinetics and translocation sites for the PKC isozymes α and δ upon treatment with the compounds were studied as well as the early response of ERK phosphorylation and the late response of induction of apoptosis in the human prostatic carcinoma cell line LNCaP. It was found that the different spatial disposition of the rigid structural motif on the DAG-lactones contributed to differential activity.