基诺美
激酶
生物化学
苏氨酸
丝氨酸
天然产物
丝氨酸苏氨酸激酶
药物发现
生物
蛋白激酶A
IC50型
活动站点
高通量筛选
磷酸化
表型筛选
化学
酶
体外
基因
表型
作者
Lin Du,Brice A P Wilson,Ning Li,Rohan Shah,Masoumeh Dalilian,Dongdong Wang,Emily A. Smith,Antony Wamiru,Ekaterina I. Goncharova,Ping Zhang,Barry R O'Keefe
标识
DOI:10.1021/acs.jnatprod.3c00394
摘要
The DNAJB1-PRKACA oncogenic gene fusion results in an active kinase enzyme, J-PKAcα, that has been identified as an attractive antitumor target for fibrolamellar hepatocellular carcinoma (FLHCC). A high-throughput assay was used to identify inhibitors of J-PKAcα catalytic activity by screening the NCI Program for Natural Product Discovery (NPNPD) prefractionated natural product library. Purification of the active agent from a single fraction of an Aplidium sp. marine tunicate led to the discovery of two unprecedented alkaloids, aplithianines A (1) and B (2). Aplithianine A (1) showed potent inhibition against J-PKAcα with an IC50 of ∼1 μM in the primary screening assay. In kinome screening, 1 inhibited wild-type PKA with an IC50 of 84 nM. Further mechanistic studies including cocrystallization and X-ray diffraction experiments revealed that 1 inhibited PKAcα catalytic activity by competitively binding to the ATP pocket. Human kinome profiling of 1 against a panel of 370 kinases revealed potent inhibition of select serine/threonine kinases in the CLK and PKG families with IC50 values in the range ∼11-90 nM. An efficient, four-step total synthesis of 1 has been accomplished, enabling further evaluation of aplithianines as biologically relevant kinase inhibitors.
科研通智能强力驱动
Strongly Powered by AbleSci AI