Five novel modified linear peptides named brintonamides A–E (1–5) were discovered from a marine cyanobacterial sample collected from Brinton Channel, Florida Keys. The total synthesis of 1–5 in addition to two other structurally related analogues (6 and 7) was achieved, which provided more material to allow rigorous biological evaluation and SAR studies. Compounds were subjected to cancer-focused phenotypic cell viability and migration assays and orthogonal target-based pharmacological screening platforms to identify their protease and GPCR modulatory activity profiles. The cancer related serine protease kallikrein 7 (KLK7) was inhibited to similar extents with an IC₅₀ near 20 μM by both representative members 1 and 4, which differed in the presence or lack of the N-terminal unit. In contrast to the biochemical protease profiling study, clear SAR was observed in the functional GPCR screens, where five GPCRs in antagonist mode (CCR10, OXTR, SSTR3, TACR2) and agonist mode (CXCR7) were modulated by compounds 1–7 to varying extents. Chemokine receptor type 10 (CCR10) was potently modulated by brintonamide D (4) with an IC₅₀ of 0.44 μM. We performed in silico modeling to understand the structural basis underlying the differences in the antagonistic activity among brintonamides toward CCR10. Because of the significance of KLK7 and CCR10 in cancer progression and metastasis, we demonstrated the ability of brintonamide D (4) at 10 μM to significantly target downstream cellular substrates of KLK7 (Dsg-2 and E-cad) in vitro and to inhibit CCL27-induced CCR10-mediated proliferation and the migration of highly invasive breast cancer cells.

中文翻译：

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从海洋蓝细菌中发现，合成，药理学概况和Brintonamides AE，新型双重蛋白酶和GPCR调节剂的生物学特性。

改性的线性肽命名brintonamides A-E（5个新的1 - 5）是从由布林顿频道，佛罗里达收集海洋蓝细菌样品发现的。的全合成1 - 5除了其他两种结构上相关的类似物（6和7）中的溶液来实现，其提供了更多的材料，以允许严格生物学评价和SAR研究。对化合物进行了针对癌症的表型细胞生存力和迁移测定，以及基于正交靶标的药理筛选平台，以鉴定其蛋白酶和GPCR调节活性谱。用IC ₅₀将与癌症相关的丝氨酸蛋白酶激肽释放酶7（KLK7）抑制至相似程度两个代表性成员1和4接近20μM ，这在N末端单元的存在与否方面有所不同。与此相反的生化蛋白酶谱的研究中，明确SAR在功能GPCR屏幕，观察到其中拮抗剂模式（CCR10，OXTR，SSTR3，TACR2）和激动剂模式（CXCR7）通过化合物调制5周的GPCR 1 - 7在不同程度上。布雷宁酰胺D（4）用IC ₅₀有效调节10型趋化因子受体（CCR10）。为0.44μM。我们进行了计算机模拟，以了解布雷宁酰胺对CCR10拮抗活性之间差异的基础。由于KLK7和CCR10在癌症进展和转移中的重要意义，我们证明了在10μM浓度的brintonamide D（4）在体外显着靶向KLK7的下游细胞底物（Dsg-2和E-cad）并抑制CCL27的能力。诱导的CCR10介导的增殖和高侵袭性乳腺癌细胞的迁移。