Design, synthesis and activity of a potent, selective series of N-aryl pyridinone inhibitors of p38 kinase

Bioorganic and Medicinal Chemistry Letters

Volumn 21, Issue 13, 2011, Pages 4059-4065

  Selness, Shaun R ^a   Boehm, Terri L ^a   Walker, John K ^a   Devadas, Balekudru ^a   Durley, Richard C ^a   Kurumbail, Ravi ^a   Shieh, Huey ^a   Xing, Li ^a   Hepperle, Michael ^a   Rucker, Paul V ^a   Jerome, Kevin D ^a   Benson, Alan G ^a   Marrufo, Laura D ^a   Madsen, Heather M ^a   Hitchcock, Jeff ^a   Owen, Tom J ^a   Christie, Lance ^a   Promo, Michele A ^a   Hickory, Brian S ^a   Alvira, Edgardo ^a   Naing, Win ^a   Blevis Bal, Radhika ^a   Devraj, Rajesh V ^a   Messing, Dean ^a   Schindler, John F ^a   Hirsch, Jeffrey ^a   Saabye, Matthew ^a   Bonar, Sheri ^a   Webb, Elizabeth ^a   Anderson, Gary ^a   Monahan, Joseph B ^a   more..

^a PFIZER INC   (United States)

Author keywords

Inflammation; p38 kinase; Pyridinones; Rheumatoid arthritis; TNF

Indexed keywords

2 BROMOETHANOL; 5 (2,6 DICHLOROPHENYL) 2 (2,4 DIFLUOROPHENYLTHIO)PYRIMIDO[1,6 B]PYRIDAZIN 6 ONE; BENZYL DERIVATIVE; HYDROXY ACETAMIDE DERIVATIVE; METHYLAMINE; MITOGEN ACTIVATED PROTEIN KINASE P38 INHIBITOR; N PHENYL 3 BROMO 4 BENZYLOXY PYRIDINONE; N PHENYL 3 CHLORO 4 BENZYLOXY; PHENYL GROUP; PYRIDONE DERIVATIVE; SC 25028; UNCLASSIFIED DRUG;

ANIMAL CELL; ANIMAL EXPERIMENT; ANIMAL MODEL; ARTICLE; CONTROLLED STUDY; DISEASE SEVERITY; DRUG ABSORPTION; DRUG ACTIVITY; DRUG BIOAVAILABILITY; DRUG DESIGN; DRUG POTENCY; DRUG SCREENING; DRUG STABILITY; DRUG STRUCTURE; DRUG SYNTHESIS; HUMAN; HUMAN CELL; INFLAMMATION; MALE; NONHUMAN; RAT; SUBSTITUTION REACTION;

ANIMALS; DISEASE MODELS, ANIMAL; DRUG DESIGN; ENZYME ACTIVATION; ENZYME INHIBITORS; HUMANS; INHIBITORY CONCENTRATION 50; MALE; MICROSOMES, LIVER; MOLECULAR STRUCTURE; P38 MITOGEN-ACTIVATED PROTEIN KINASES; PYRIDAZINES; PYRIDONES; PYRIMIDINES; RATS; RATS, SPRAGUE-DAWLEY;

EID: 79958714435     PISSN: 0960894X     EISSN: 14643405     Source Type: Journal    
DOI: 10.1016/j.bmcl.2011.04.120     Document Type: Article

References (19)

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12. Full disclosure of earlier assay methods is made in the following patent application: Devadas, Balekudru; Walker, John; Selness, Shaun, R.; Boehm, Terri L.; Durley, Richard C.; Devraj, Rajesh; Hickory, Brian S.; Rucker, Paul V.; Jerome, Kevin D.; Madsen, Heather M.; Alvira, Edgardo; Promo, Michele A.; Blevis-Bal, Radhika M.; Marrufo, Laura D.; Hitchcock, Jeff; Owen, Thomas; Naing, Win; Xing, Li; Shieh, Huey S.; Sambandam, Aruna; Liu, Shuang; Scott, Ian L.; Mcgee, Kevin F. PCT Int. Appl. 2005, 968, pp. WO 2005018557.
13. 2, 0.0005% Tween-20, 0.01% BSA, 1 mM DTT, and 2% DMSO). The inhibitors were varied between 0.2 and 10,000 nM, while the Hsp27 peptide substrate, MgATP, and unactivated MK2 were held constant at 1, 10, and 1 nM, respectively. Reactions were initiated by the addition of activated p38α to a final concentration of 6 pM. Kinase reactions were incubated at room temperature and quenched after 60 minutes by the addition of stop buffer (180 mM HEPES, 30 mM EDTA, and 0.2% Coating Reagent-3).
14. 2SO concentration in cell assay was 0.25%. Secreted TNF-α was measured by MSD technology (MSD, Gaithersburg, MD). IC50s were determined using an internal data analysis program (Pfizer, St. Louis).
15. Metabolic stability was assessed in vitro by incubating 2 μM test compound with human or rat liver microsomes, NADPH and buffer at 37°C for 45 min and measuring percent compound remaining by a precipitation procedure followed by LC/MS analysis.
16. Adult male Lewis rats (Harlan Sprague Dawley, Indianapolis, IN) (225-250 g) were used in these studies. Rats were fasted 18 h prior to oral dosing, and allowed free access to water throughout the experiment. Each treatment group consisted of five animals. 10 was prepared as a suspension in a vehicle consisting of 0.5% methylcellulose, (Sigma, St. Louis, MO), 0.025% Tween 20 (Sigma). The compound or vehicle was administered by oral gavage in a volume of 1 mL. Two vehicle groups were used per experiment to control for intra-experiment variability, and three experiments were performed. LPS (E. coli serotype 0111:B4, Sigma) was administered two, four or six hours later by intravenous injection at a dose of 1 mg/kg in 0.5 mL sterile saline (Baxter Healthcare, Deerfield, IL). Blood was collected in serum separator tubes via cardiac puncture ninety minutes after LPS injection, a time point corresponding to maximal TNFα production (data not shown). After clotting, serum was withdrawn and stored at -20°C until it was assayed for TNFα. TNFα levels in serum were quantified from a recombinant rat TNFα (Biosource International) standard curve using a four parameter fit generated by an Excel (Microsoft, Redmond, WA) macro. The limit of detection for the ELISA was approximately 41 pg TNFα/mL.
17. The Pfizer Institutional Animal Care and Use Committee reviewed and approved the animal use in these studies. The animal care and use program is fully accredited by the Association for Assessment and Accreditation of Laboratory Animal Care, International.
18. Compounds 2 and 3 assayed in Streptococal Cell Wall (SCW) induced arthritis in rats as follows: Arthritis was induced in female Lewis rats by a single intraperitoneal administration of peptidoglycan-polysaccaride complexes isolated from group a SCW (15 μg/g bodyweight). The SCW preparation was purchase from Lee Labs (Grayson, GA). The disease course is biphasic in which an acute inflammatory arthritis develops within days 1-3 (nonT-cell-dependent phase) followed by a chronic erosive arthritis (T-cell- dependent phase) developing on days 14-28. Only animals developing the acute phase were treated with compound from days 10 to 21 after SCW injection. Paw volume was measured on day 21 by using a water displacement plethysmometer. Each compound was prepared as an aqueous suspension in 0.5% methylcellulose and 0.025% Tween 20(sigma-aldrich). The compound was administered by oral gavage in a volume of 0.5 mL beginning on day 10 post-SCW injection and continuing daily until day 21. Methylcellulose/Tween 20 vehicle was used for comparison. Group size was four to eight animals per group. Two paw volumes were taken for each animal. Paw volume was measured on day 21 by using a water displacement plethysmometer. Three to four paws from each treatment group were scanned for bone density evaluation. Plasma samples were collected on day 21 for determination of compound levels.
19. Compounds 2 and 3 exhibited less than 50% at 10 μM against over 200 targets including, but not limited to, the following kinases: BTK, CDK2, AURKA, EGFR, FLT1, ERK1, mTOR, IKKβ, IRAK4, JAK1, JAK2, JAK3, TYK2, VEGFR2, LCK, MEK1, p38δ, p38γ, ERK1, JNK, MK-2, TRKA, PDGFR, cRAF, ROCK1, ROCK2 and ZAP70. The only significant inhibition observed was for p38α and p38β.