Cenderitide: structural requirements for the creation of a novel dual particulate guanylyl cyclase receptor agonist with renal-enhancing in vivo and ex vivo actions
Cenderitide: structural requirements for the creation of a novel dual particulate guanylyl cyclase receptor agonist with renal-enhancing in vivo and ex vivo actions
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DOI：
10.1093/ehjcvp/pvv040
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发表时间：
2016-04-01
影响因子：
7.1
通讯作者：
Burnett, John C., Jr.
中科院分区：
文献类型：
--
作者：
Lee, Candace Y. W.;Huntley, Brenda K.;Burnett, John C., Jr.
Cenderitide 是一种新型双利尿钠肽 (NP) 受体嵌合肽激活剂，与天然 NP 不同，它靶向颗粒鸟苷酸环化酶 B (pGC-B) 受体和 pGC-A。 Cenderitide 经过改造，保留了 C 型利钠肽 (CNP)/pGC-B 的抗纤维化特性，并通过与 Dendroaspis NP (DNP)（一种 pGC-A 激动剂）的羧基末端融合来促进肾增强作用。在这里，我们讨论了 DNP 羧基末端在双 pGC 受体激活中的重要性以及 cenderitide 与 CNP 对正常犬体内和离体肾功能和环磷酸鸟苷 (cGMP) 的作用相比。在体外，只有 cenderitide 而不是 CNP 或三个基于 CNP 的变体是人胚胎肾中 cGMP 产生（从 5 至 237 pmol/mL）的有效双 pGC-A/pGC-B 激活剂(HEK) 293 细胞过表达人 pGC-A，而在 pGC-B 过表达细胞中 cenderitide 增加了 cGMP 产量（从 4 至 321 pmol/mL），而三种基于 CNP 的变体是弱激动剂。基于我们发现 DNP 羧基末端是 pGC-A/pGC-B 双激活的关键结构要求，我们定义了 cenderitide 与 CNP 相比的体内肾脏增强作用。 Cenderitide 增加尿 cGMP 排泄（从 989 至 5977 pmol/mL）、肾 cGMP 净生成（821-4124 pmol/min）、尿钠（12-242 mu Eq/min）和肾小球滤过率（GFR）（37-51 mL/min），而 CNP 则没有。然后，我们证明了 CNP 离体转化为肾 cGMP 激活肽，其使新鲜分离的肾小球中的 cGMP 增加了 CNP 的八倍。当前的研究表明，CNP 的双重 pGC-A 和 pGC-B 激活需要 DNP 的特定羧基末端。在体内正常犬科动物和离体肾小球中，DNP 的羧基末端将 CNP 转化为利尿钠和 GFR 增强肽。未来需要在心肾疾病状态下研究 cenderitide，以探索其在整体心肾稳态中的功效。
Cenderitide is a novel dual natriuretic peptide (NP) receptor chimeric peptide activator, which targets the particulate guanylyl cyclase B (pGC-B) receptor and pGC-A unlike native NPs. Cenderitide was engineered to retain the anti-fibrotic properties of C-type natriuretic peptide (CNP)/pGC-B with renal-enhancing actions facilitated by fusion to the carboxyl terminus of Dendroaspis NP (DNP), a pGC-A agonist, to CNP. Here, we address significance of the DNP carboxyl terminus in dual pGC receptor activation and actions of cenderitide compared with CNP on renal function and cyclic guanosine monophosphate (cGMP) in vivo and ex vivo in normal canines.In vitro, only cenderitide and not CNP or three CNP-based variants was a potent dual pGC-A/pGC-B activator of cGMP production (from 5 to 237 pmol/mL) in human embryonic kidney (HEK) 293 cells overexpressing human pGC-A while in pGC-B overexpressing cells cenderitide increased cGMP production (from 4 to 321 pmol/mL) while the three CNP-based variants were weak agonists. Based upon our finding that the DNP carboxyl terminus is a key structural requirement for dual pGC-A/pGC-B activation, we defined in vivo the renal-enhancing actions of cenderitide compared with CNP. Cenderitide increased urinary cGMP excretion (from 989 to 5977 pmol/mL), net generation of renal cGMP (821-4124 pmol/min), natriuresis (12-242 mu Eq/min), and glomerular filtration rate (GFR) (37-51 mL/min) while CNP did not. We then demonstrated the transformation of CNP ex vivo into a renal cGMP-activating peptide which increased cGMP in freshly isolated glomeruli eight-fold greater than CNP.The current study establishes that dual pGC-A and pGC-B activation with CNP requires the specific carboxyl terminus of DNP. In normal canines in vivo and in glomeruli ex vivo, the carboxyl terminus of DNP transforms CNP into a natriuretic and GFR-enhancing peptide. Future studies of cenderitide are warranted in cardiorenal disease states to explore its efficacy in overall cardiorenal homeostasis.