IC50型
肽
分子生物学
噬菌体展示
癌基因
酶
生长抑制
磷酸化
细胞生长
重组DNA
激酶
癌症研究
癌症
MAPK/ERK通路
癌细胞
生物
化学
细胞
细胞培养
生物化学
细胞周期
基因
遗传学
作者
Kotaro Sakamoto,Yusuke Kamada,Tomoya Sameshima,Masahiro Yaguchi,Ayumu Niida,Shigekazu Sasaki,Masanori Miwa,Shoichi Ohkubo,Daniel J. Sargent,Masahiro Kamaura,Nobuo Cho,Akiyoshi Tani
标识
DOI:10.1016/j.bbrc.2017.01.147
摘要
Amino-acid mutations of Gly12 (e.g. G12D, G12V, G12C) of V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (K-Ras), the most promising drug target in cancer therapy, are major growth drivers in various cancers. Although over 30 years have passed since the discovery of these mutations in most cancer patients, effective mutated K-Ras inhibitors have not been marketed. Here, we report novel and selective inhibitory peptides to K-Ras(G12D). We screened random peptide libraries displayed on T7 phage against purified recombinant K-Ras(G12D), with thorough subtraction of phages bound to wild-type K-Ras, and obtained KRpep-2 (Ac-RRCPLYISYDPVCRR-NH2) as a consensus sequence. KRpep-2 showed more than 10-fold binding- and inhibition-selectivity to K-Ras(G12D), both in SPR analysis and GDP/GTP exchange enzyme assay. KD and IC50 values were 51 and 8.9 nM, respectively. After subsequent sequence optimization, we successfully generated KRpep-2d (Ac-RRRRCPLYISYDPVCRRRR-NH2) that inhibited enzyme activity of K-Ras(G12D) with IC50 = 1.6 nM and significantly suppressed ERK-phosphorylation, downstream of K-Ras(G12D), along with A427 cancer cell proliferation at 30 μM peptide concentration. To our knowledge, this is the first report of a K-Ras(G12D)-selective inhibitor, contributing to the development and study of K-Ras(G12D)-targeting drugs.
科研通智能强力驱动
Strongly Powered by AbleSci AI