6-Ethynylthieno[3,2-d]- and 6-ethynylthieno[2,3-d]pyrimidin-4-anilines as tunable covalent modifiers of ErbB kinases.
Analysis of the x-ray crystal structure of mono-substituted acetylenic thienopyrimidine 6 complexed with the ErbB family enzyme ErbB-4 revealed a covalent bond between the terminal carbon of the acetylene moiety and the sulfhydryl group of Cys-803 at the solvent interface. The identification of this covalent adduct suggested that acetylenic thienopyrimidine 6 and related analogs might also be capable of forming an analogous covalent adduct with EGFR, which has a conserved cysteine (797) near the ATP binding pocket. To test this hypothesis, we treated a truncated, catalytically competent form of EGFR (678-1020) with a structurally related propargylic amine (8). An investigation of the resulting complex by mass spectrometry revealed the formation of a covalent complex of thienopyrimidine 8 with Cys-797 of EGFR. This finding enabled us to readily assess the irreversibility of various inhibitors and also facilitated a structure-activity relationship understanding of the covalent modifying potential and biological activity of a series of acetylenic thienopyrimidine compounds with potent antitumor activity. Several ErbB family enzyme and cell potent 6-ethynyl thienopyrimidine kinase inhibitors were found to form covalent adducts with EGFR.
Wood
Edgar R
ER
Department of Assay Development, GlaxoSmithKline, Research Triangle Park, NC 27709, USA.
Shewchuk
Lisa M
LM
Ellis
Byron
B
Brignola
Perry
P
Brashear
Ronald L
RL
Caferro
Thomas R
TR
Dickerson
Scott H
SH
Dickson
Hamilton D
HD
Donaldson
Kelly H
KH
Gaul
Michael
M
Griffin
Robert J
RJ
Hassell
Anne M
AM
Keith
Barry
B
Mullin
Robert
R
Petrov
Kimberly G
KG
Reno
Michael J
MJ
Rusnak
David W
DW
Tadepalli
Sarva M
SM
Ulrich
John C
JC
Wagner
Craig D
CD
Vanderwall
Dana E
DE
Waterson
Alex G
AG
Williams
Jon D
JD
White
Wendy L
WL
Uehling
David E
DE
D000480:Alkynes / Q000378:metabolism*; D000814:Aniline Compounds / Q000378:metabolism*; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D018360:Crystallography, X-Ray; D004305:Dose-Response Relationship, Drug; D066246:ErbB Receptors / Q000378:metabolism*; D005260:Female; D007510:Isatin / Q000031:analogs & derivatives / Q000378:metabolism; D013058:Mass Spectrometry; D051379:Mice; D016513:Mice, SCID; D008958:Models, Molecular; D015394:Molecular Structure; D011743:Pyrimidines / Q000378:metabolism* / Q000633:toxicity; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism; D013329:Structure-Activity Relationship; D023041:Xenograft Model Antitumor Assays
D000480:Alkynes; D000814:Aniline Compounds; D011743:Pyrimidines; C116442:propargylic sulfone; C476003:thienopyrimidine; D007510:Isatin; D066246:ErbB Receptors; D020794:Receptor Protein-Tyrosine Kinases