title: Bisarylureas Based on 1H-Pyrazolo[3,4-d]pyrimidine Scaffold as Novel Pan-RAF Inhibitors with Potent Anti-Proliferative Activities: Structure-Based Design, Synthesis, Biological Evaluation and Molecular Modelling Studies.
abstract: RAF (Ras activating factor) kinases are important and attractive targets for cancer therapy. With the aim of discovering RAF inhibitors that bind to DFG-out inactive conformation created by the movement of Asp-Phe-Gly (DFG), we conducted structure-based drug design using the X-ray cocrystal structures of BRAF (v-raf murine sarcoma viral oncogene homolog B1), starting from bisarylurea derivative based on 1H-pyrazolo[3,4-d]pyrimidine scaffold1a. Most of the synthesized compounds showed good to excellent inhibitory activities against BRAFV600Ekinase, possessed moderate to potent anti-proliferative activities against four tumor cell lines (A375, HT-29, PC-3 and A549) and good selectivity towards cancer cells rather normal cells (Madin-Darby canine kidney, MDCK). The most promising compound,1v, exhibited potent inhibitory activity against not only BRAFV600E(half maximal inhibitory concentration, IC50= 23.6 nM) but also wild-type BRAF (IC50= 51.5 nM) and C-RAF (IC50= 8.5 nM), and effective cellular anti-proliferative activities against A375, HT-29, PC-3 and A549 cell lines as well as a very good selectivity profile. Moreover, compound1vmainly arrested the A375 cell line in the G0/G1 stage, and showed significant suppression of MEK (mitogen-activated protein kinase kinase) phosphorylation in A375 and HT-29 cell lines. Taken together, the optimal compound1vshowed excellent in vitro potency as a pan-RAF inhibitor. In addition, the promise of compound1vwas further confirmed by molecular dynamics simulation and binding free energy calculations.
authors.affiliation: Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Science, Southern Medical University, Guangzhou 510515, China. fuyuky0312@163.com.
authors.forename: Yu
authors.initials: Y
authors.lastname: Fu
authors.affiliation: Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Science, Southern Medical University, Guangzhou 510515, China. 15626047153@163.com.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Science, Southern Medical University, Guangzhou 510515, China. wansh@fimmu.com.
authors.forename: Shanhe
authors.initials: S
authors.lastname: Wan
authors.affiliation: Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Science, Southern Medical University, Guangzhou 510515, China. lzhuang@fimmu.com.
authors.forename: Zhonghuang
authors.initials: Z
authors.lastname: Li
authors.affiliation: Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Science, Southern Medical University, Guangzhou 510515, China. wguangfa@smu.edu.cn.
authors.forename: Guangfa
authors.initials: G
authors.lastname: Wang
authors.affiliation: Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Science, Southern Medical University, Guangzhou 510515, China. zhangjj@smu.edu.cn.
authors.forename: Jiajie
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Science, Southern Medical University, Guangzhou 510515, China. xywugz@163.com.
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Wu
mesh_terms: D000072283:A549 Cells; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D015195:Drug Design; D004354:Drug Screening Assays, Antitumor; D019073:HT29 Cells; D006801:Humans; D008958:Models, Molecular; D047428:Protein Kinase Inhibitors / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D048493:Proto-Oncogene Proteins B-raf / Q000037:antagonists & inhibitors* / Q000737:chemistry; D011743:Pyrimidines / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship
chemical_list: D047428:Protein Kinase Inhibitors; D011743:Pyrimidines; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf
keywords: 1H-pyrazolo[3,4-d]pyrimidine derivatives; DFG-out; biological activities; molecular docking; molecular dynamics simulation; pan-RAF inhibitor