Use of pyrido2,3-bpyrazin-6-yl urea derivatives for preparation of medicaments for treating diseases mediated by inhibition of pi3k enzyme

摘要

Treatment and prevention of physiological and pathophysiological state mediated by signal transduction pathway selected from rat sarcoma-rapid accelerated fibrosarcoma-mitogen activated protein kinase-extracellualr signal-regulated kinase signal transduction pathway, phosphoinositol 3-kinase-Akt signal transduction pathway and stress-activated protein kinase signal transduction pathway comprises administering a pyridopyrazine derivative (I). Treatment or prevention of physiological and/or pathophysiological state mediated by at least one signal transduction pathway selected from rat sarcoma-rapid accelerated fibrosarcoma-mitogen activated protein kinase-extracellualr signal-regulated kinase signal transduction pathway, the phosphoinositol 3-kinase-Akt signal transduction pathway and the stress-activated protein kinase signal transduction pathway comprises administering a pyridopyrazine derivatives of formula (I). R 1and R 2 : e.g. aryl (optionally substituted by T 1) or heteroaryl (optionally substituted by T 2); T 1 : e.g. F or heteroaryl; T 2 : e.g. F or NH-alkyl-NH 2; R 3and R 4 : H or NR 9R 10; R 9 : e.g. alkyl or alkyl-heteroaryl (all optionally substituted) or H; R 10 : -C(Y 1)NR 11R 1) 2; Y 1 : O or S; R 11and R 12 : alkyl (optionally substituted by e.g. T 3or SO 2O-alkyl-aryl), cycloalkyl (optionally substituted by e.g. F or aryl), heterocyclyl (optionally substituted by e.g. OH or aryl), aryl (optionally substituted by T 1or S-heterocyclyl), heteroaryl (optionally substituted by T 2) or H, and T 3 : e.g. CO 2H or heteroaryl. Full Definitions are given in the DEFINITIONS (Full Definitions) section. Independent claims are also included for: (1) new pyridopyrazine derivatives (I') selected from 232 compounds as given in the specification e.g. 1-ethyl-3-[3-(3-hydroxy-phenyl)-pyrido[2,3-b]pyrazin-6-yl]-urea; (2) a pharmaceutical composition (C1) comprising compounds (I'), and (3) a kit comprising compounds (I') and active substance. [Image] ACTIVITY : Cytostatic; Antiinflammatory; Analgesic; Antirheumatic; Antiarthritic; Anti-HIV; Neuroprotective; Nootropic; Antipsoriatic; Gynecological; Vulnerary; Immunosuppressive; Vasotropic; Antidiabetic; Nephrotropic; Anticoagulant; Thrombolytic; Ophthalmological; Antiasthmatic; Antiallergic; Respiratory-Gen.; Gastrointestinal-Gen.; Antiarteriosclerotic; Cardiant; Cardiovascular-Gen.; Antithyroid; Cerebroprotective; Anorectic; Antianginal; Hypotensive; Angiogenesis inhibitor. The efficacy of 1-ethyl-3-[3-p-tolylamino-pyrido[2,3-b]pyrazin-6-yl]-urea (Ia) was evaluated for cytostatic activity using sodium 3'-[1-(phenylaminocarbonyl)-3,4-tetrazolium]-bis(4-methoxy-6-nitro)benzenesulfonic acid (XTT) assay. The tumor cell lines were injected into 96-well microtitre plates and then incubated overnight in an incubator at 37[deg]C, 5% carbon dioxide (CO 2) and 95% air humidity. (Ia) (0.28 mu M) was added to the cells in quarter-logarithmically graded dilutions. The cell plates were then incubated for 45 hours in an incubator at 37[deg]C, 5% CO 2and 95% air humidity and EC 50value measured. (Ia) Showed an EC 50value of 0.9 mu M. MECHANISM OF ACTION : Extracellular signal-regulated kinase inhibitor; Phosphoinositol 3-kinase inhibitor; Stress-activated protein kinase inhibitor. The efficacy of 1-ethyl-3-[3-(3-hydroxy-phenyl)-pyrido[2,3-b]pyrazin-6-yl]-urea (Ib) was evaluated for extracellular signal-regulated kinase inhibitory activity using extracellular signal-regulated kinase assay. 1-Ethyl-3-[3-(3-hydroxyphenyl)-pyrido[2,3-b]pyrazin-6-yl]urea (Ib), extracellular signal-regulated kinase (0.625 ng), adenosine triphosphate (10 mu M)and myelin basic protein (MBP) substrate (15 nM) were incubated on a 384-well Optiplate in a volume of 15 mu l for 1 hour in Tris (25 mM), magnesium dichloride (10 mM), Tween 20(RTM: Polysorbate 20) (0.1%), NaVO 4(100 mu M), DTT (2 mM) at pH 7.5. The kinase reaction was then stopped by adding the bead mixes (10 mu l) pre-incubated with anti-phospho MBP antibody (320 pM), in Tris (25 mM), sodium chloride (200 mM), ethylenediaminetetraacetic acid (110 nM) and bovine serum albumin (0.3%) and IC 50value measured. (Ib) Showed IC 50of 0.104 mu M.