Structure-activity relationship studies of pyrazolo[3,4-d]pyrimidine derivatives leading to the discovery of a novel multikinase inhibitor that potently inhibits FLT3 and VEGFR2 and evaluation of its activity against acute myeloid leukemia in vitro and in vivo

Ling-Ling Yang; Guo-Bo Li; Shuang Ma; Chan Zou; Shu Zhou; Qi-Zheng Sun; Chuan Cheng; Xin Chen; Li-Jiao Wang; Shan Feng; Lin-Li Li; Sheng-Yong Yang

doi:10.1021/jm301537p

Structure-activity relationship studies of pyrazolo[3,4-d]pyrimidine derivatives leading to the discovery of a novel multikinase inhibitor that potently inhibits FLT3 and VEGFR2 and evaluation of its activity against acute myeloid leukemia in vitro and in vivo

J Med Chem. 2013 Feb 28;56(4):1641-55. doi: 10.1021/jm301537p. Epub 2013 Feb 19.

Authors

Ling-Ling Yang¹, Guo-Bo Li, Shuang Ma, Chan Zou, Shu Zhou, Qi-Zheng Sun, Chuan Cheng, Xin Chen, Li-Jiao Wang, Shan Feng, Lin-Li Li, Sheng-Yong Yang

Affiliation

¹ State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Sichuan 610041, China.

PMID: 23362959
DOI: 10.1021/jm301537p

Abstract

We describe the structural optimization of a hit compound, 1-(4-(1H-pyrazolo[3,4-d]pyrimidin-4-ylamino)phenyl)-3-(3-methoxyphenyl)urea (1), which exhibits inhibitory activity but low potency against FLT3 and VEGFR2. A series of pyrazolo[3,4-d]pyrimidine derivatives were synthesized, and structure-activity relationship analysis using cell- and transgenic-zebrafish-based assays led to the discovery of a number of compounds that exhibited both high potency against FLT3-driven human acute myeloid leukemia (AML) MV4-11 cells and a considerable antiangiogenic effect in transgenic-zebrafish-based assays. The compound 1-(4-(1H-pyrazolo[3,4-d]pyrimidin-4-yloxy)phenyl)-3-(4-chloro-3-(trifluoromethyl)phenyl)urea (33), which exhibited the highest activity in preliminary in vivo anti-AML assays, was chosen for further anti-AML studies. The results demonstrated that compound 33 is a multikinase inhibitor that potently inhibits FLT3 and VEGFR2. In an MV4-11 xenograft mouse model, a once-daily dose of compound 33 at 10 mg/kg for 18 days led to complete tumor regression without obvious toxicity. Western blot and immunohistochemical analyses were performed to determine the mechanism of action of compound 33.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Angiogenesis Inhibitors / chemical synthesis
Angiogenesis Inhibitors / chemistry
Angiogenesis Inhibitors / pharmacology
Animals
Animals, Genetically Modified
Antineoplastic Agents / chemical synthesis*
Antineoplastic Agents / chemistry
Antineoplastic Agents / pharmacology
Apoptosis / drug effects
Cell Line, Tumor
Cell Proliferation / drug effects
Drug Screening Assays, Antitumor
Embryo, Nonmammalian / blood supply
Embryo, Nonmammalian / drug effects
Humans
Leukemia, Myeloid, Acute / drug therapy*
Leukemia, Myeloid, Acute / pathology
Mice
Neoplasm Transplantation
Phenylurea Compounds
Pyrazoles / chemical synthesis*
Pyrazoles / chemistry
Pyrazoles / metabolism
Pyrazoles / pharmacology
Pyrimidines / chemical synthesis*
Pyrimidines / chemistry
Pyrimidines / metabolism
Pyrimidines / pharmacology
Signal Transduction / drug effects
Structure-Activity Relationship
Transplantation, Heterologous
Urea / analogs & derivatives*
Urea / chemical synthesis
Urea / chemistry
Urea / metabolism
Vascular Endothelial Growth Factor Receptor-2 / antagonists & inhibitors*
Zebrafish
fms-Like Tyrosine Kinase 3 / antagonists & inhibitors*
fms-Like Tyrosine Kinase 3 / metabolism

Substances

1-(4-(1H-pyrazolo(3,4-d)pyrimidin-4-yloxy)phenyl)-3-(4-chloro-3-(trifluoromethyl)phenyl)urea
Angiogenesis Inhibitors
Antineoplastic Agents
Phenylurea Compounds
Pyrazoles
Pyrimidines
Urea
FLT3 protein, human
Vascular Endothelial Growth Factor Receptor-2
fms-Like Tyrosine Kinase 3