4 research outputs found
Site-Selective Protein Immobilization by Covalent Modification of GST Fusion Proteins
The immobilization of functional
proteins onto solid supports using
affinity tags is an attractive approach in recent development of protein
microarray technologies. Among the commonly used fusion protein tags,
glutathione <i>S</i>-transferase (GST) proteins have been
indispensable tools for protein–protein interaction studies
and have extensive applications in recombinant protein purification
and reversible protein immobilization. Here, by utilizing pyrimidine-based
small-molecule probes with a sulfonyl fluoride reactive group, we
report a novel and general approach for site-selective immobilization
of Schistosoma japonicum GST (<i>sj</i>GST) fusion proteins through irreversible and specific
covalent modification of the tyrosine-111 residue of the <i>sj</i>GST tag. As demonstrated by <i>sj</i>GST-tagged eGFP and <i>sj</i>GST-tagged kinase activity assays, this immobilization
approach offers the advantages of high immobilization efficiency and
excellent retention of protein structure and activity
Discovery of a Series of 2,5-Diaminopyrimidine Covalent Irreversible Inhibitors of Bruton’s Tyrosine Kinase with in Vivo Antitumor Activity
Bruton’s tyrosine kinase (Btk)
is an attractive drug target for treating several B-cell lineage cancers.
Ibrutinib is a first-in-class covalent irreversible Btk inhibitor
and has demonstrated impressive effects in multiple clinical trials.
Herein, we present a series of novel 2,5-diaminopyrimidine covalent
irreversible inhibitors of Btk. Compared with ibrutinib, these inhibitors
exhibited a different selectivity profile for the analyzed kinases
as well as a dual-action mode of inhibition of both Btk activation
and catalytic activity, which counteracts a negative regulation loop
for Btk. Two compounds from this series, <b>31</b> and <b>38</b>, showed potent antiproliferative activities toward multiple
B-cell lymphoma cell lines, including germinal center B-cell-like
diffuse large B cell lymphoma (GCB-DLBCL) cells. In addition, compound <b>31</b> significantly prevented tumor growth in a mouse xenograft
model
Selective Inhibitors of Human Neuraminidase 3
Human
neuraminidases (NEU) are associated with human diseases including
cancer, atherosclerosis, and diabetes. To obtain small molecule inhibitors
as research tools for the study of their biological functions, we
designed a library of 2-deoxy-2,3-didehydro-<i>N</i>-acetylneuraminic
acid (DANA) analogues with modifications at C4 and C9 positions. This
library allowed us to discover selective inhibitors targeting the
human NEU3 isoenzyme. Our most selective inhibitor for NEU3 has a <i>K</i><sub>i</sub> of 320 ± 40 nM and a 15-fold selectivity
over other human neuraminidase isoenzymes. This inhibitor blocks glycolipid
processing by NEU3 in vitro. To improve their pharmacokinetic properties,
various esters of the best inhibitors were synthesized and evaluated.
Finally, we confirmed that our best compounds exhibited selective
inhibition of NEU orthologues from murine brain
Selective Inhibitors of Human Neuraminidase 3
Human
neuraminidases (NEU) are associated with human diseases including
cancer, atherosclerosis, and diabetes. To obtain small molecule inhibitors
as research tools for the study of their biological functions, we
designed a library of 2-deoxy-2,3-didehydro-<i>N</i>-acetylneuraminic
acid (DANA) analogues with modifications at C4 and C9 positions. This
library allowed us to discover selective inhibitors targeting the
human NEU3 isoenzyme. Our most selective inhibitor for NEU3 has a <i>K</i><sub>i</sub> of 320 ± 40 nM and a 15-fold selectivity
over other human neuraminidase isoenzymes. This inhibitor blocks glycolipid
processing by NEU3 in vitro. To improve their pharmacokinetic properties,
various esters of the best inhibitors were synthesized and evaluated.
Finally, we confirmed that our best compounds exhibited selective
inhibition of NEU orthologues from murine brain