Biphenyl amide p38 kinase inhibitors 3: Improvement of cellular and in vivo activity

Bioorganic and Medicinal Chemistry Letters

Volumn 18, Issue 15, 2008, Pages 4428-4432

  Angell, Richard ^a   Aston, Nicola M ^a   Bamborough, Paul ^a   Buckton, Jacky B ^a   Cockerill, Stuart ^a   deBoeck, Suzanne J ^a   Edwards, Chris D ^a   Holmes, Duncan S ^a   Jones, Katherine L ^a   Laine, Dramane I ^a   Patel, Shila ^a   Smee, Penny A ^a   Smith, Kathryn J ^a   Somers, Don O ^a   Walker, Ann L ^a  

^a GLAXOSMITHKLINE   (United Kingdom)

Author keywords

Binding mode; Biphenyl amide; Biphenyl amides; BPA; BPAs; CIA model; CSBP; Inhibitors; Kinase selectivity; MAP kinase; p38; p38 Alpha; p38 Kinase; PG PS model; Protein kinase X ray structure; Structure activity relationships; Structure based drug design

Indexed keywords

ANILIDE; BENZAMIDE DERIVATIVE; BIPHENYLAMIDE; MITOGEN ACTIVATED PROTEIN KINASE 14; MITOGEN ACTIVATED PROTEIN KINASE P38; MITOGEN ACTIVATED PROTEIN KINASE P38 ALPHA INHIBITOR; MITOGEN ACTIVATED PROTEIN KINASE P38 INHIBITOR; UNCLASSIFIED DRUG;

ANIMAL EXPERIMENT; ARTICLE; CELL ACTIVITY; DRUG ACTIVITY; DRUG POTENCY; IN VIVO STUDY; NONHUMAN; RAT; STRUCTURE ACTIVITY RELATION; X RAY CRYSTALLOGRAPHY;

ADMINISTRATION, ORAL; ANIMALS; BENZAMIDES; BIPHENYL COMPOUNDS; COMBINATORIAL CHEMISTRY TECHNIQUES; CRYSTALLOGRAPHY, X-RAY; DISEASE MODELS, ANIMAL; MITOGEN-ACTIVATED PROTEIN KINASE 14; MOLECULAR CONFORMATION; MOLECULAR STRUCTURE; PROTEIN KINASE INHIBITORS; RATS;

EID: 47749150617     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2008.06.048     Document Type: Article

References (15)

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2. Angell R.M., Bamborough P., Cleasby A., Cockerill S., Jones K.L., Mooney C.J., Somers D.O., and Walker A.L. Bioorg. Med. Chem. Lett. 18 (2008) 318
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4. Angell, R.; Aston, N.M.; Bamborough, P.; Buckton, J.B.; Cockerill, S., de Boeck, S.J.; Edwards, C.D.; Holmes, D.S.; Jones, K.L.; Laine, D.I.; Patel, S.; Smee, P.A.; Smith, K.J.; Somers, D.O.; Walker, A.L.; Bioorg. Med. Chem. Lett., accepted for publication.
5. i determination, using an assay measuring displacement of a fluorescent ATP-competitive inhibitor was carried out as described in reference 1.
6. 2 X-ray diffraction data were collected from the crystal at 100 K (using an Oxford Cryostream) on a Rigaku-MSC RuH2R rotating anode X-ray generator with a RAXIS IV++ image-plate detector. The data were processed with the HKL package (Otwinowski, Z.; Minor, W. Methods Enzymol. 1997, 276:Macromol. Crys. A, 307-326) and CCP4 program suite (Bailey, S. Acta Crystallogr., Sect. D, 1994, 50, 760-763). The structure was solved using the native p38 coordinates (PDB entry 1WFC) as the initial model in refinement by REFMAC (Murshudov, G.; Vagin, A.; Dodson, E. Acta Crystallogr., Sect. D, 1997, D53, 240-255). The final R-factor achieved for the complex was 17.4%. Coordinates have been deposited in the PDB as entry 3D7Z.
7. Harrison Jr. J.H., and Hollow D.J. J. Pharmacol. Exp. Ther. 238 (1986) 1045
8. Osazaki Y., Yamashita K., Ishii H., Sudi M., and Tsuchitani M. J. Appl. Toxicol. 23 (2003) 315
9. m)).
10. Assays were carried out by Ambit Biosciences, San Diego. The ability of compounds to compete with the binding of the human kinase (expressed as fusion to T7 bacteriophage) to immobilized ATP-site probe ligands was determined as previously described, see:. Fabian M.A., Biggs III W.H., Treiber D.K., Atteridge C.E., Azimioara M.D., Benedetti M.G., Carter T.A., Ciceri P., Edeen P.T., Floyd M., Ford J.M., Galvin M., Gerlach J.L., Grotzfeld R.M., Herrgard S., Insko D.E., Insko M.A., Lai A.G., Lélias J.-M., Mehta S.A., Milanov Z.V., Velasco A.M., Wodicka L.M., Patel H.K., Zarrinkar P.P., and Lockhart D.J. Nat. Biotechnol. 23 (2005) 329 Compounds were screened at 10 μM and considered active if <10% of binding to immobilised probes remained compared to DMSO control.
11. Goldstein D.M., and Gabriel T. Curr. Topics Med. Chem. 5 (2005) 1017
12. 50 determination assay was 0.1-100 μM (spread over 9 points) and included a 0.2 mM NADPH regeneration system.
13. 2 for 20 h, 100-150 μl physiological saline (0.138% NaCl) was added and diluted plasma was collected using a Platemate or Biomek FX liquid handling robot after centrifugation at 1300g for 10 min. Plasma TNFα content was determined by enzyme linked immunosorbant assay (ELISA) or using a multiplex bead technology (Luminex).
14. Pharmacokinetic parameters in male Lewis rats were determined following intravenous (iv) and oral (po) administration at 0.5 mg/kg and 1.5 mg/kg, respectively. Compound was administered as a solution in 10% DMSO: 45% SBE Cyclodextrin, 45% MSA (IV) or 5% DMSO, 40% vitE, 40% PEG200, 15% Mannitol (po). Blood was collected over a 24-h time period. Plasma was prepared following centrifugation and compound extracted from 50 μl plasma using protein precipitation with acetonitrile. Samples were then evaporated under nitrogen and re-suspended in 100 μl of 10:90 acetonitrile/water. Analysis was performed using LC-MSMS on the API365 with a 5 min fast gradient comprising 0.1% formic acid in water and 0.1% formic acid in acetonitrile (mobile phases), 20 μl injection volume, flow rate 4 ml/min and ODS3 Prodigy column (5 cm × 2.1 mm, 5 μm). Pharmacokinetic data was generated using a non-compartmental approach. The brain penetration was assessed in the Lewis rat, collecting brains and terminal blood samples five minutes after an intravenous administration at 1 mg/kg, using the same formulation as in the intravenous PK study. Brains were homogenised in a water:methanol 1:1 solution and the homogenates analysed with a protein precipitation method similar to the one detailed for plasma analysis.
15. CIA is a widely used animal model of arthritis in which the anti-inflammatory and anti-rheumatic efficacy of drugs and novel compounds predicted to have activity in RA are evaluated. This model is one of the more widely used animal models of RA and shares many similarities with the human disease, for example synovial hyperplasia, infiltration of inflammatory cells, erosion of cartilage and bone and involvement of both B and T lymphocytes. It has extensively been reported in the literature that CIA can be inhibited by anti-cytokine reagents (IL-1b, IL-6 and to a lesser extent TNFα). CIA was induced by sensitising DBA/1 mice against bovine type II collagen + Freund's Complete adjuvant injected intradermally at the base of the tail followed 3 weeks later by an intraperitoneal booster injection of bovine type II collagen. The activity of compound 3 in this model was assessed by orally dosing mice twice daily for 14 days with 15 or 30 mg/kg starting when the mice were showing early signs of inflammation in the paws. Enbrel (soluble TNFα receptor) was tested in the same study as a comparator. Clinical scores were monitored throughout the dosing period as a measure of anti-inflammatory effect and at the end of the study, ankle joints were processed histologically to assess effects on structural damage.