Selective targeting of the inactive state of hematopoietic cell kinase (Hck) with a stable curcumin derivative

J Biol Chem. 2021 Jan-Jun:296:100449. doi: 10.1016/j.jbc.2021.100449. Epub 2021 Feb 20.

Abstract

Hck, a Src family nonreceptor tyrosine kinase (SFK), has recently been established as an attractive pharmacological target to improve pulmonary function in COVID-19 patients. Hck inhibitors are also well known for their regulatory role in various malignancies and autoimmune diseases. Curcumin has been previously identified as an excellent DYRK-2 inhibitor, but curcumin's fate is tainted by its instability in the cellular environment. Besides, small molecules targeting the inactive states of a kinase are desirable to reduce promiscuity. Here, we show that functionalization of the 4-arylidene position of the fluorescent curcumin scaffold with an aryl nitrogen mustard provides a stable Hck inhibitor (Kd = 50 ± 10 nM). The mustard curcumin derivative preferentially interacts with the inactive conformation of Hck, similar to type-II kinase inhibitors that are less promiscuous. Moreover, the lead compound showed no inhibitory effect on three other kinases (DYRK2, Src, and Abl). We demonstrate that the cytotoxicity may be mediated via inhibition of the SFK signaling pathway in triple-negative breast cancer and murine macrophage cells. Our data suggest that curcumin is a modifiable fluorescent scaffold to develop selective kinase inhibitors by remodeling its target affinity and cellular stability.

Keywords: Hck inhibitor; Src family kinase; cell signaling; curcumin derivative; enzyme kinetics; kinase inhibition.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cloning, Molecular
  • Curcumin / analogs & derivatives
  • Curcumin / chemical synthesis
  • Curcumin / pharmacology*
  • Drug Design*
  • Drug Stability
  • Dyrk Kinases
  • Epithelial Cells / drug effects*
  • Epithelial Cells / enzymology
  • Epithelial Cells / pathology
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Gene Expression Regulation
  • Genetic Vectors / chemistry
  • Genetic Vectors / metabolism
  • HEK293 Cells
  • HT29 Cells
  • Humans
  • Mice
  • Mitogen-Activated Protein Kinase 1 / genetics
  • Mitogen-Activated Protein Kinase 1 / metabolism
  • Mitogen-Activated Protein Kinase 3 / genetics
  • Mitogen-Activated Protein Kinase 3 / metabolism
  • Protein Kinase Inhibitors / chemical synthesis
  • Protein Kinase Inhibitors / pharmacology*
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism
  • Protein-Tyrosine Kinases / genetics
  • Protein-Tyrosine Kinases / metabolism
  • Proto-Oncogene Proteins c-abl / genetics
  • Proto-Oncogene Proteins c-abl / metabolism
  • Proto-Oncogene Proteins c-hck / antagonists & inhibitors*
  • Proto-Oncogene Proteins c-hck / chemistry
  • Proto-Oncogene Proteins c-hck / genetics
  • Proto-Oncogene Proteins c-hck / metabolism
  • RAW 264.7 Cells
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Structure-Activity Relationship
  • src-Family Kinases / genetics
  • src-Family Kinases / metabolism

Substances

  • Protein Kinase Inhibitors
  • Recombinant Proteins
  • Protein-Tyrosine Kinases
  • HCK protein, human
  • Proto-Oncogene Proteins c-abl
  • Proto-Oncogene Proteins c-hck
  • src-Family Kinases
  • Protein Serine-Threonine Kinases
  • MAPK1 protein, human
  • MAPK3 protein, human
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3
  • Curcumin