Acta Chimica Sinica ›› 2013, Vol. 71 ›› Issue (08): 1161-1166.DOI: 10.6023/A13030320 Previous Articles     Next Articles



侯雪艳a, 武锋a, 周孟a, 罗浩a, 张文娟a, 韩璇a, 闫国毅a, 师健友b, 李锐a   

  1. a 四川大学 生物治疗国家重点实验室 成都 610041;
    b 四川省医学科学院·四川省人民医院 药剂科 成都 610072
  • 投稿日期:2013-03-21 发布日期:2013-05-16
  • 通讯作者: 师健友;李锐, E-mail:; Tel.: 028-85164063; Fax: 028-85164060
  • 基金资助:

    项目受国家自然科学基金(No. 30901743)、教育部博士点新教师基金(No. 20090181120113)以及国家科技重大专项(No. 2012ZX09103101-022)资助.

Study of Fragmentation Mechanisms of Pyrrolo[3,4-c]pyrazole Derived Aurora Kinase Inhibitors by ESI-QTOF-MS/MS

Hou Xueyana, Wu Fenga, Zhou Menga, Luo Haoa, Zhang Wenjuana, Han Xuana, Yan Guoyia, Shi Jianyoub, Li Ruia   

  1. a State Key Laboratory of Biotherapy, Sichuan University, Chengdu 610041;
    b Department of Pharmacy, Sichuan Academy of Medical Science and Sichuan Provincial People's Hospital, Chengdu 610072
  • Received:2013-03-21 Published:2013-05-16
  • Supported by:

    Project supported by the National Natural Science Foundation of China (No. 30901743), the Doctoral Fund of Ministry of Education (No. 20090181120113) and the National Key Program of China (No. 2012ZX09103101-022).

Aurora kinase inhibitors represent an emerging class of drugs against various tumors such as acute lymphoblastic leukemia and hepatocellular carcinomain. A variety of drug candidates have been developed and investigated including a group that comprises pyrrolo[3,4-c]pyrazole core such as danusertib (PHA-739358). Due to their novelty and medicinal purpose, four pyrrolo[3,4-c]pyrazole derivatives were synthesized and were conducted to provide information on typical fragmentation pathways by electrospray ionization (ESI) and collision induced dissociation (CID). The product ions derived from protonated molecules were investigated by ESI-quadrupole time of flight tandem mass spectrometry (QTOF-MS/MS) and theoretical methods. It should be pointed that four favorite protonation modes were observed and each mode had its own characteristic fragmentation pathways. Major and general findings contained the elimination of R2 group from[M+H]+and the dissociation of C(4)—N(5), C(6)—N(5) bonds, which was depended on the most favorable protonation mode, while the other product ions were formed based on the other protonation modes. Interestingly, the loss of 18 Da from potential aurora kinase inhibitors 3, 4 was occurred, even though there was no free hydroxyl group. Accurate mass measurements have allowed us to determine that a molecule of water was eliminated from protonated molecules directly. This process may start with the most favorable protonation mode; then a conformational change implied by rotation of the acetyl group around the internal 3-amide bond of protonated molecules occurred to yield an intermediate. Followed by the hydrogen transference and elongation of C—O bond, a molecule of water was expelled to give the product ion[M+H-H2O]+. The process was supported by the theoretical calculations. Additionally, two other special product ions were formed according to the hydration reaction. It was viewed that the origin of H2O was the elimination of protonated molecules. These observations may provide some applications for monitoring and characterization of their presence and metabolites in complex mixtures, such as urine and blood.

Key words: aurora kinase inhibitors, pyrrolo[3,4-c]pyrazole derivatives, ESI-QTOF-MS/MS, fragmentation mechanisms, isomers