Stefanucci Azzurra, Lei Wei, Hruby Victor J, Macedonio Giorgia, Luisi Grazia, Carradori Simone, Streicher John M, Mollica Adriano
Dipartimento di Farmacia, Università di Chieti-Pescara "G. d'Annunzio", Via dei Vestini 31, 66100 Chieti, Italy.
Department of Pharmacology, College of Medicine, University of Arizona, 85721 AZ, USA.
Future Med Chem. 2017 Jun;9(9):859-869. doi: 10.4155/fmc-2016-0232. Epub 2017 Jun 21.
The conjugation of fluorescent labels to opioid peptides is an extremely challenging task, which needs to be overcome to create new classes of probes for biological assays.
MATERIALS & METHODS: Three opioid peptide analogs of biphalin and [D-Pen2,5]-Enkephalin (DPDPE) containing a fluorescein-maleimide motif were synthesized.
RESULTS & DISCUSSION: The biphalin analog 17 binds to opioid receptors with K = 530 ± 90 nM and K = 69.8 ± 16.4 nM. We then tested the ability of the compounds to stimulate G-protein-coupling, 17 activated μ-receptor expressing cells (EC = 16.7 ± 6.7 nM, E = 76 ± 4%) as well as δ-receptor expressing cells (EC = 42 ± 10 nM, E = 34 ± 8%). However, 17 was not able to fluorescently label receptor in live or fixed cells.
Our data suggest that the biphalin scaffold could be employed to develop fluorescent ligands with the appropriate fluorescent motif, and suggest a means for further probe development.
将荧光标记与阿片肽偶联是一项极具挑战性的任务，要创建用于生物测定的新型探针就必须克服这一难题。
合成了三种含有荧光素 - 马来酰亚胺基序的双氢吗啡肽和[D - 青霉胺2,5] - 脑啡肽（DPDPE）的阿片肽类似物。
双氢吗啡肽类似物17与阿片受体结合，其K值分别为530±90 nM和69.8±16.4 nM。然后我们测试了这些化合物刺激G蛋白偶联的能力，17激活了表达μ受体的细胞（EC = 16.7±6.7 nM，E = 76±4%）以及表达δ受体的细胞（EC = 42±10 nM，E = 34±8%）。然而，17无法在活细胞或固定细胞中对受体进行荧光标记。
我们的数据表明，双氢吗啡肽支架可用于开发具有合适荧光基序的荧光配体，并为进一步的探针开发提供了一种方法。
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