Π-Clamp-mediated cysteine conjugation.
Π-Clamp-mediated cysteine conjugation.
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DOI：
10.1038/nchem.2413
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发表时间：
2016-02
期刊：
影响因子：
21.8
通讯作者：
Pentelute BL
中科院分区：
文献类型：
--
作者：
Zhang C;Welborn M;Zhu T;Yang NJ;Santos MS;Van Voorhis T;Pentelute BL
复杂分子的位点选择性功能化是化学领域的一个重大挑战。必须使用保护基团或催化剂来选择性地修饰许多类似反应性的位点中的一个位点。一般的策略是罕见的，这样的目标位点周围的局部化学环境被调整为选择性转化。在这里，我们展示了一个四个氨基酸的序列（Phe-Cys-Pro-Phe），我们称之为“π-钳”，调整其半胱氨酸巯基的反应性，用于与全氟芳香族试剂的位点选择性缀合。我们使用π-钳来选择性地修饰含有多个内源性半胱氨酸残基的蛋白质（例如抗体和基于半胱氨酸的酶）中的一个半胱氨酸位点，这在先前的半胱氨酸修饰方法中是不可能的。修饰的π-钳抗体保留了对其靶标的结合亲和力，使得能够合成位点特异性抗体-药物缀合物（ADC）以选择性杀死HER 2阳性乳腺癌细胞。π-钳是用于位点选择性化学的意想不到的方法，并且提供了修饰用于研究和治疗的生物分子的机会。
Site-selective functionalization of complex molecules is a grand challenge in chemistry. Protecting groups or catalysts must be used to selectively modify one site among many that are similarly reactive. General strategies are rare such the local chemical environment around the target site is tuned for selective transformation. Here we show a four amino acid sequence (Phe-Cys-Pro-Phe), which we call the “π-clamp”, tunes the reactivity of its cysteine thiol for the site-selective conjugation with perfluoroaromatic reagents. We used the π-clamp to selectively modify one cysteine site in proteins containing multiple endogenous cysteine residues (e.g. antibodies and cysteine-based enzymes), which was impossible with prior cysteine modification methods. The modified π-clamp antibodies retained binding affinity to their targets, enabling the synthesis of site-specific antibody-drug conjugates (ADCs) for selective killing of HER2-positive breast cancer cells. The π-clamp is an unexpected approach for site-selective chemistry and provides opportunities to modify biomolecules for research and therapeutics.