Advances in Tetrazine Bioorthogonal Chemistry Driven by the Synthesis of Novel Tetrazines and Dienophiles.
Advances in Tetrazine Bioorthogonal Chemistry Driven by the Synthesis of Novel Tetrazines and Dienophiles.
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DOI：
10.1021/acs.accounts.8b00062
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发表时间：
2018-05-15
影响因子：
18.3
通讯作者：
Devaraj NK
中科院分区：
文献类型：
--
作者：
Wu H;Devaraj NK
在过去的十年里，生物正交化学在生命系统中的应用越来越多。特别是，四嗪生物正交化学已成为一个强大的工具，成像，检测和诊断的目的，反映在文献中报道的例子越来越多。四嗪连接的流行可能是由于快速和可调的动力学，高质量的荧光探针的存在，以及反应的选择性。在这个帐户中，我们总结了我们最近的努力，以推进四嗪生物正交化学通过改进合成方法，重点是开发新的路线，以四嗪和扩大范围的有用的亲二烯体。这些努力消除了以前限制四嗪连接的特定障碍，并扩大了其潜在的应用。在其他进展中，这个帐户描述了我们的小组如何发现新的方法，四嗪合成通过开发一个刘易斯酸促进，一锅法生成不同的对称和不对称的烷基四嗪与功能取代基在令人满意的产率。我们将这些四嗪连接到微电极上，并成功地通过改变反应物的氧化还原状态来控制四嗪连接。使用这种电化学控制过程，我们能够以位点选择性方式用氧化还原探针和酶修饰电极表面。这个帐户还描述了我们的小组如何提高四嗪作为荧光探针的能力，通过开发一种新的消除赫克级联反应合成烯基四嗪衍生物。在该方法中，将四嗪缀合至荧光团以产生强淬灭的探针，在生物正交反应之后，所述探针被“打开”以增强荧光，在许多情况下增强>100倍。这些探针允许在活细胞和完整动物中进行免清洗荧光成像。最后，这个帐户审查我们的努力，扩大范围的亲二烯体基板，使四嗪生物正交化学与特定的生物化学和生物医学应用兼容。我们发现，甲基环丙烯是足够的稳定性和反应性的生物环境中作为一个有效的亲二烯体。反应性标签的小尺寸使空间位阻最小化，允许环丙烯充当代谢报告基团以揭示生物动力学和功能。我们还使用降冰片二烯衍生物作为紧张的亲二烯体来进行四嗪介导的转移（TMT）反应，该反应涉及四嗪连接，然后是逆狄尔斯-桤木过程。该TMT反应产生一对非连接产物。使用核酸模板化化学，我们将TMT反应与我们的荧光四嗪探针相结合，以检测皮摩尔浓度的内源性致癌microRNA。在亲二烯体多功能性的进一步展示中，我们使用新型乙烯基醚以非荧光形式笼住近红外荧光团。然后，我们在“点击释放”四嗪生物正交反应中打开笼子，恢复荧光团的荧光形式。将该标记与相应的核酸探针组合允许靶mRNA的荧光检测。总之，该帐户描述了四嗪和亲二烯体合成和应用的改进，以推进四嗪生物正交化学。这些进展进一步使四嗪连接化学不仅在基础研究中而且在诊断研究中的应用成为可能。
Bioorthogonal chemistry has found increased application in living systems over the past decade. In particular, tetrazine bioorthogonal chemistry has become a powerful tool for imaging, detection, and diagnostic purposes, as reflected in the increased number of examples reported in the literature. The popularity of tetrazine ligations are likely due to rapid and tunable kinetics, the existence of high quality fluorogenic probes, and the selectivity of reaction. In this Account, we summarize our recent efforts to advance tetrazine bioorthogonal chemistry through improvements in synthetic methodology, with an emphasis on developing new routes to tetrazines and expanding the range of useful dienophiles. These efforts have removed specific barriers that previously limited tetrazine ligations and have broadened their potential applications. Among other advances, this Account describes how our group discovered new methodology for tetrazine synthesis by developing a Lewis acid-promoted, one-pot method for generating diverse symmetric and asymmetric alkyl tetrazines with functional substituents in satisfactory yields. We attached these tetrazines to microelectrodes and succeeded in controlling tetrazine ligation by changing the redox state of the reactants. Using this electrochemical control process, we were able to modify an electrode surface with redox probes and enzymes in a site-selective fashion. This Account also describes how our group improved the ability of tetrazines to act as fluorogenic probes by developing a novel elimination-Heck cascade reaction to synthesize alkenyl tetrazine derivatives. In this approach, tetrazine was conjugated to fluorophores to produce strongly quenched probes that, after bioorthogonal reaction, are “turned on” to enhance fluorescence, in many cases by >100-fold. These probes have allowed no-wash fluorescence imaging in living cells and intact animals. Finally, this Account reviews our efforts to expand the range of dienophile substrates to make tetrazine bioorthogonal chemistry compatible with specific biochemical and biomedical applications. We found that methylcyclopropene is sufficiently stable and reactive in the biological milieu to act as an efficient dienophile. The small size of the reactive tag minimizes steric hindrance, allowing cyclopropene to serve as a metabolic reporter group to reveal biological dynamics and function. We also used norbornadiene derivatives as strained dienophiles to undergo tetrazine-mediated transfer (TMT) reactions involving tetrazine ligation followed by a retro-Diels–Alder process. This TMT reaction generates a pair of nonligating products. Using nucleic acid-templated chemistry, we have combined the TMT reaction with our fluorogenic tetrazine probes to detect endogenous oncogenic microRNA at picomolar concentrations. In a further display of dienophile versatility, we used a novel vinyl ether to cage a near-infrared fluorophore in a nonfluorescent form. Then we opened the cage in a “click to release” tetrazine bioorthogonal reaction, restoring the fluorescent form of the fluorophore. Combining this label with a corresponding nucleic acid probe allowed fluorogenic detection of target mRNA. In summary, this Account describes improvements in tetrazine and dienophile synthesis and application to advance tetrazine bioorthogonal chemistry. These advances have further enabled application of tetrazine ligation chemistry, not only in fundamental research but also in diagnostic studies.
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影响因子：
15
作者：
Blackman, Melissa L.;Royzen, Maksim;Fox, Joseph M.
通讯作者：
Fox, Joseph M.
DOI：
10.1002/anie.201403349
发表时间：
2014-09-15
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Erdmann RS;Takakura H;Thompson AD;Rivera-Molina F;Allgeyer ES;Bewersdorf J;Toomre D;Schepartz A
通讯作者：
Schepartz A
影响因子：
15
作者：
Cohen AS;Dubikovskaya EA;Rush JS;Bertozzi CR
通讯作者：
Bertozzi CR
影响因子：
3.2
作者：
Cole, Christian M.;Yang, Jun;Seckute, Jolita;Devaraj, Neal K.
通讯作者：
Devaraj, Neal K.
影响因子：
--
作者：
Hofmann, KA;Ehrhart, O
通讯作者：
Ehrhart, O