The development of 2-benzimidazole substituted pyrimidine based inhibitors of lymphocyte specific kinase (Lck)

Bioorganic and Medicinal Chemistry Letters

Volumn 16, Issue 23, 2006, Pages 5973-5977

  Sabat, Mark ^a   VanRens, John C ^a   Laufersweiler, Matthew J ^a   Brugel, Todd A ^a   Maier, Jennifer ^a   Golebiowski, Adam ^a   De, Biswanath ^a   Easwaran, Vijayasurian ^a   Hsieh, Lily C ^a   Walter, Richard L ^b   Mekel, Marlene J ^b   Evdokimov, Artem ^b   Janusz, Michael J ^a  

^a PROCTER AND GAMBLE PHARMACEUTICALS   (United States)

^b Miami Valley Innovation Center   (United States)

Author keywords

Hck; Hematopoietic cell kinase; Kinase; Lck; Lymphocyte specific kinase; Src family kinase; T cell

Indexed keywords

2 BENZIMIDAZOLECARBAMIC ACID DERIVATIVE; ENZYME INHIBITOR; PROTEIN KINASE LCK; PROTEIN KINASE LCK INHIBITOR; PYRIMIDINE DERIVATIVE; UNCLASSIFIED DRUG;

ANIMAL CELL; ARTICLE; CRYSTALLIZATION; DRUG ACTIVITY; DRUG BINDING; DRUG CLEARANCE; DRUG METABOLISM; DRUG POTENCY; DRUG SOLUBILITY; DRUG SYNTHESIS; ENZYME INHIBITION; HUMAN; HUMAN CELL; NONHUMAN; RAT; SUBSTITUTION REACTION;

BENZIMIDAZOLES; HYDROGEN BONDING; INHIBITORY CONCENTRATION 50; INTERLEUKIN-2; LYMPHOCYTE SPECIFIC PROTEIN TYROSINE KINASE P56(LCK); MOLECULAR STRUCTURE; PROTEIN KINASE INHIBITORS; PROTO-ONCOGENE PROTEINS C-HCK; PYRIMIDINES; STRUCTURE-ACTIVITY RELATIONSHIP;

EID: 33750531634     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2006.08.132     Document Type: Article

References (26)

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11. 50 value. The assay Quality Control Criteria inlcuded; Signal/Background: >4; Signal/Noise: >10; % Coefficient of Variance on No ATP Control (Max. Signal): <10.
12. 50) was determined using the commercially available ProFlour Src-family Kinase Assay (Promega Corporation, Madison, WI; cat. #1271). The assay was performed according to manufacturer's instructions, with 2.8 nM recombinant human active Hck (Invitrogen, Carlsbad, CA #P2908) at an ATP concentration of 10 mM.
13. 50) was determined using the commercially available ProFlour Src-family Kinase Assay (Promega Corporation, Madison, WI; cat. #1271). The assay was performed according to manufacturer's instructions, with 14.3 nM recombinant human active Src (Invitrogen, Carlsbad, CA #P3044) at an ATP concentration of 10 mM.
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19. 6 cells/ml) were harvested and incubated in triplicate in 96-well plates for 30 min at 37 °C in the presence or absence of various concentrations of Lck inhibitors. The cell-inhibitor mixture was then transferred into the wells of the anti CD3ε-coated 96-well plates, and PMA was added to the wells at a final concentration of 10 ng/ml (1 ng/well). The plates were incubated overnight at 37 °C. The amount of IL-2 released into the culture media was measured by ELISA (R&D Systems) and the viability of the cells was determined using the MTS assay (Promega, Madison, WI).
20. Measured as percent loss at 4 hours in rat hepatocytes.
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26. The non-phos Hck construct used in these studies appears to place the activation loop in a conformation resembling activated Lck (3LCK, 1QPC; phosphorylated Y394); the authors speculate inhibitor 12k is capable of inducing such a conformation through interaction with Glu288.