Synthesis and evaluation of bifunctional sGC regulators: optimization of a connecting linker.
Synthesis and evaluation of bifunctional sGC regulators: optimization of a connecting linker.
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双功能 sGC 调节剂的合成和评估：连接子的优化。
DOI：
10.1021/jm400715h
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发表时间：
2013
影响因子：
7.3
通讯作者：
Martin,Emil
中科院分区：
文献类型：
--
作者：
Chromiński,Mikołaj;Banach,Łukasz;Karczewski,Maksymilian;óProinsias,Keith;Sharina,Iraida;Gryko,Dorota;Martin,Emil
通过1，3-偶极环加成（CuAAC）或酰胺化/酯化反应制备了由PpIX和cobyrinic酸衍生物通过不同长度和组成的连接体缀合组成的杂合分子。通过体外GTP→cGMP转化试验，检测它们对可溶性鸟苷酸环化酶（sGC）（NO/cGMP信号通路中的关键酶）的活化。使用纯化的血红素缺陷型sGC和缺乏血红素结合结构域的截短型sGC变体，我们证明了这样的杂合分子可以通过靶向血红素结合和/或催化结构域来激活sGC。虽然所有缀合物都激活sGC，但仅选定的化合物充当双功能调节剂，并且能够同时靶向sGC的血红素和催化结构域。连接两种组分的接头的长度和类型对sGC活化的程度具有深远的影响，表明接头的类型对于它们与调节和催化结构域的结合亲和力至关重要。只有具有13-16个原子长度的缀合接头的杂合体协同靶向两个结构域，并显示最低的EC 50和最高的活化效力。具有较短连接接头的化合物的效力低得多，并且不比单独的钴啉酸组分更有活性。显示出60倍sGC活化的最有活性的缀合物是化合物11，其中PpIX和钴啉酸组分被11个原子链隔开，三唑部分在其间。
Hybrid molecules composed of PpIX and cobyrinic acid derivatives conjugated through linkers of varying length and composition were prepared via 1,3-dipolar cycloaddition (CuAAC) or amidation/esteryfication reactions. They were tested for activation of soluble guanylyl cyclase (sGC), a key enzyme in the NO/cGMP signaling pathway, by an in vitro GTP→cGMP conversion assay. Using purified heme-deficient sGC and truncated sGC variants lacking a heme-binding domain, we demonstrated that such hybrid molecules may activate sGC by targeting heme-binding and/or catalytic domain. While all conjugates activated sGC, only selected compounds served as bifunctional regulators and were capable of simultaneous targeting both heme and catalytic domains of sGC. The length and type of a linker connecting both components had a profound effect on the extent of sGC activation, indicating that the linker’s type is crucial for their binding affinities with regulatory and catalytic domains. Only hybrids with the conjugated linker of 13–16 atom length synergistically target both domains and displayed the lowest EC50and highest activating potency. Compounds with shorter connecting linkers were much less potent and were no more active than the cobyrinic acid component alone. The most active conjugate, which showed a 60-fold activation of sGC, was compound11, in which PpIX and cobyrinic acid components are separated by 11 atoms chain with the triazole moiety in between.