Generation of monospecific nanomolar tyrosine kinase inhibitors via a chemical genetic approach
Generation of monospecific nanomolar tyrosine kinase inhibitors via a chemical genetic approach
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DOI：
10.1021/ja983267v
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发表时间：
1999-02-03
影响因子：
15
通讯作者：
Liu, Y
中科院分区：
文献类型：
--
作者：
Bishop, AC;Kung, CY;Liu, Y
选择性蛋白激酶抑制剂作为研究细胞信号转导级联的工具而受到高度追捧，但由于激酶催化结构域的高度保守折叠，很少被发现。通过小分子合成和蛋白质诱变的结合，已鉴定出合理设计的 v-Src 酪氨酸激酶 (Ile338Gly v-Src) 的高效 (IC50 = 1.5 nM) 和独特特异性抑制剂 (4-氨基-1-叔丁基-3-(1'-萘基)吡唑并[3,4-d]嘧啶)。该化合物的效力和特异性均超过任何已知的 Src 家族酪氨酸激酶抑制剂。该分子强烈抑制全细胞中的工程化 v-Src，但不抑制仅表达野生型酪氨酸激酶的细胞中的酪氨酸磷酸化。此外，该抑制剂选择性地破坏表达靶标 v-Src 的细胞的转化。激酶活性位点的结构简并性应允许相同的互补抑制剂/蛋白质设计策略广泛适用于整个酶超家族。
Selective protein kinase inhibitors are highly sought after as tools for studying cellular signal transduction cascades, yet few have been discovered due to the highly conserved fold of kinase catalytic domains. Through a combination of small molecule synthesis and protein mutagenesis, a highly potent (IC50 = 1.5 nM) and uniquely specific inhibitor (4-amino-1-tert-butyl-3-(1'-naphthyl)pyrazolo[3,4-d]pyrimidine) of a rationally engineered v-Src tyrosine kinase (Ile338Gly v-Src) has been identified. Both the potency and specificity of this compound surpass those of any known Src family tyrosine kinase inhibitors. The molecule strongly inhibits the engineered v-Src in whole cells but does not inhibit tyrosine phosphorylation in cells that express only wild-type tyrosine kinases. In addition, the inhibitor selectively disrupts transformation in cells that express the target v-Src. The structural degeneracy of kinase active sites should allow the same complementary inhibitor/protein design strategy to be widely applicable across this entire enzyme superfamily.