RESUMO
BACKGROUND: Evaluate the potential role of p38 inhibitors for the treatment of osteoarthritis using an animal model of joint degeneration (iodoacetate-induced arthritis) and a pain model (Hargraeves assay). METHODS: P38 kinase activity was evaluated in a kinase assay by measuring the amount of phosphorylated substrate ATF2 using a phosphoATF2 (Thr71) specific primary antibody and an alkaline phosphate coupled secondary antibody and measuring the OD at 405 nm. TNFalpha and IL-1beta secretion from LPS stimulated THP-1 monocytic cells and human peripheral blood mononuclear cells were measured by ELISA. Rats treated with vehicle or p38 inhibitor were injected intra-articularly in one knee with iodoacetate and damage to the tibial plateau was assessed from digitized images captured using an image analyzer. The effect of p38 inhibitors on hyperalgesia was evaluated in rats given an intraplantar injection of carrageenan and 4 h later the paw withdrawal time to a radiant heat source was measured. RESULTS: SB-203580 and VX-745 are both potent inhibitors of p38 with IC50s of 136 +/- 64 nM and 35 +/- 14 nM (mean +/- S.D.), respectively. Similarly, SB-203580 and VX-745 potently inhibited TNF release from LPS stimulated human THP-1 cells with IC50s of 72 +/- 15 nM; and 29 +/- 14 nM (mean +/- S.D.) respectively. TNF release from LPS stimulated human peripheral blood mononuclear cells was inhibited with IC50s 16 +/- 6 nM and 14 +/- 8 nM, (mean +/- S.D.) for SB-203580 and VX-745 and IL-1 was inhibited with IC50s of 20 +/- 8 nM and 15 +/- 4 nM (mean +/- S.D.), respectively. SB-203580 and VX-745 administered orally at a dose of 50 mg/kg resulted in the significant (p < 0.05) inhibition of joint degeneration in the rat iodoacetate model of 45% and 31%, respectively. SB-203580 demonstrated a dose related inhibition of joint degeneration of 30, 25, 12 and 8% at 50, 25, 10 and 5 mg/kg p.o. b.i.d. in the rat iodoacetate model. Similarly, both p38 inhibitors significantly (p < 0.05) attenuated the pain response (paw withdrawal time) in the Hargraeves hyperalgesia assay when administered orally at 30, 10 and 3 mg/kg. CONCLUSION: SB203580 and VX-745 demonstrated attenuation of both cartilage degeneration and pain in animal models and suggest that p38 inhibitors may be a useful approach for the treatment of osteoarthritis.
RESUMO
This communication details the synthesis, biological activity, and binding mode of a novel class of 2-benzimidazole substituted pyrimidines. The most potent analogs disclosed showed low nanomolar activity for the inhibition of Lck kinase and a representative analog was co-crystallized with Hck (a structurally related member of the Src family kinases).
Assuntos
Benzimidazóis/química , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/antagonistas & inibidores , Inibidores de Proteínas Quinases/síntese química , Inibidores de Proteínas Quinases/farmacologia , Pirimidinas/química , Ligação de Hidrogênio , Concentração Inibidora 50 , Interleucina-2/metabolismo , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/síntese química , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/metabolismo , Estrutura Molecular , Inibidores de Proteínas Quinases/química , Proteínas Proto-Oncogênicas c-hck/química , Proteínas Proto-Oncogênicas c-hck/metabolismo , Relação Estrutura-AtividadeRESUMO
This communication details the synthesis, biological activity, and proposed binding mode of a novel class of tri-cyclic derivatives of 1,2-dihydro-pyrimido[4,5-c]pyridazines 1 and 2. The most potent analogs disclosed showed low nanomolar activity for the inhibition of Lck kinase.
Assuntos
Indústria Farmacêutica/métodos , Inibidores Enzimáticos/farmacologia , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/antagonistas & inibidores , Piridazinas/química , Pirimidinas/química , Ribonucleosídeos/química , Desenho de Fármacos , Inibidores Enzimáticos/química , Humanos , Concentração Inibidora 50 , Linfócitos/metabolismo , Modelos Químicos , Modelos Moleculares , Piridazinas/farmacologia , Pirimidinas/farmacologia , Relação Estrutura-AtividadeRESUMO
A series of C-2, C-8, and N-9 trisubstituted purine based inhibitors of TNF-alpha production are described. The most potent analogs showed low nanomolar activity against LPS-induced TNF-alpha production in a THP-1 cell based assay. The SAR of the series was optimized with the aid of X-ray co-crystal structures of these inhibitors bound with mutated p38 (mp38).
Assuntos
Purinas/química , Fator de Necrose Tumoral alfa/química , Linhagem Celular , Química Farmacêutica , Cristalografia por Raios X , Desenho de Fármacos , Humanos , Lipopolissacarídeos/química , Modelos Químicos , Modelos Moleculares , Proteínas Quinases p38 Ativadas por Mitógeno/química , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
A new class of lymphocyte specific tyrosine kinase (lck) inhibitors based on an N-4,6-pyrimidine-N-alkyl-N'-phenyl urea scaffold is described. Many of these compounds showed low-nanomolar inhibition of lck kinase activity as well as IL-2 synthesis from Jurkat cells. One of these analogs, 7i, was shown to be orally efficacious by in vivo testing in a rat adjuvant-induced arthritis study.
Assuntos
Inibidores Enzimáticos/síntese química , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/antagonistas & inibidores , Compostos de Fenilureia/síntese química , Pirimidinas/química , Administração Oral , Animais , Artrite Experimental/tratamento farmacológico , Artrite Experimental/patologia , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/uso terapêutico , Humanos , Interleucina-2/biossíntese , Células Jurkat , Estrutura Molecular , Compostos de Fenilureia/farmacologia , RatosRESUMO
A new class of tumor necrosis factor alpha (TNF-alpha) synthesis inhibitors based on a N-2,4-pyrimidine-N-phenyl-N'-alkyl urea scaffold is described. Many of these compounds showed low-nanomolar activity against lipopolysaccharide stimulated TNF-alpha production. Two analogs were tested in an in vivo rat iodoacetate model of osteoarthritis and shown to be orally efficacious. X-ray co-crystallization studies with mutated p38alpha showed that these trisubstituted ureas interact with the ATP-binding pocket in a pseudo-bicyclic conformation brought about by the presence of an intramolecular hydrogen bonding interaction.
Assuntos
Artrite Experimental/tratamento farmacológico , Osteoartrite/tratamento farmacológico , Compostos de Fenilureia/química , Compostos de Fenilureia/farmacologia , Pirimidinas/química , Pirimidinas/farmacologia , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Trifosfato de Adenosina/química , Animais , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/classificação , Anti-Inflamatórios não Esteroides/farmacologia , Artrite Experimental/induzido quimicamente , Sítios de Ligação , Cristalografia por Raios X , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Ligação de Hidrogênio , Iodoacetatos , Lipopolissacarídeos/antagonistas & inibidores , Lipopolissacarídeos/farmacologia , Modelos Moleculares , Estrutura Molecular , Osteoartrite/induzido quimicamente , Compostos de Fenilureia/classificação , Pirimidinas/classificação , Ratos , Estereoisomerismo , Relação Estrutura-Atividade , Fator de Necrose Tumoral alfa/biossínteseRESUMO
A new class of tumor necrosis factor alpha (TNF-alpha) synthesis inhibitors based on an N-2,4-pyrimidine-N-phenyl-N'-phenyl urea scaffold is described. Many of these compounds showed low-nanomolar activity against lipopolysaccharide stimulated TNF-alpha production. X-ray co-crystallization studies with mutated p38alpha showed that these trisubstituted ureas interact with the ATP-binding pocket in a pseudo-bicyclic conformation brought about by the presence of an intramolecular hydrogen bonding interaction.
Assuntos
Compostos de Fenilureia/química , Compostos de Fenilureia/farmacologia , Pirimidinas/química , Pirimidinas/farmacologia , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Trifosfato de Adenosina/química , Sítios de Ligação , Cristalografia por Raios X , Ligação de Hidrogênio , Lipopolissacarídeos/antagonistas & inibidores , Lipopolissacarídeos/farmacologia , Modelos Moleculares , Estrutura Molecular , Compostos de Fenilureia/classificação , Pirimidinas/classificação , Estereoisomerismo , Relação Estrutura-Atividade , Fator de Necrose Tumoral alfa/biossínteseRESUMO
4-Aryl-5-pyrimidyl based cytokine synthesis inhibitors that contain a novel monocyclic, pyrazolone heterocyclic core are described. Many of these inhibitors showed low nanomolar activity against LPS-induced TNF-alpha production. One of the compounds (6e) was found to be efficacious in the rat iodoacetate (RIA) in vivo model of osteoarthritis. The X-ray crystal structure of a pyrazolone inhibitor cocrystallized with mutated p38 (mp38) is presented.
Assuntos
Citocinas/antagonistas & inibidores , Citocinas/biossíntese , Pirazolonas/síntese química , Pirazolonas/farmacologia , Animais , Modelos Animais de Doenças , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/farmacologia , Cinética , Lipopolissacarídeos/farmacologia , Modelos Moleculares , Conformação Molecular , Osteoartrite/prevenção & controle , Pirazolonas/química , Ratos , Relação Estrutura-Atividade , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidoresRESUMO
4-Aryl-3-pyridyl and 4-aryl-3-pyrimidinyl based tumor necrosis factor-alpha (TNF-alpha) inhibitors, which contain a novel isoxazolone five-membered heterocyclic core are described. Many showed sub-micromolar activity against lipopolysaccharide-induced TNF-alpha production.
Assuntos
Isoxazóis/síntese química , Isoxazóis/farmacologia , Pirimidinas/síntese química , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Acetaldeído , Sítios de Ligação , Linhagem Celular , Humanos , Lipopolissacarídeos/farmacologia , Modelos Moleculares , Estrutura Molecular , Conformação Proteica , Pirimidinas/farmacologia , Relação Estrutura-Atividade , Fator de Necrose Tumoral alfa/química , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidoresRESUMO
4-Aryl-5-pyrimidyl-based cytokine synthesis inhibitors of TNF-alpha production, which contain a novel bicyclic pyrazole heterocyclic core, are described. Many of these inhibitors showed low nanomolar activity against LPS-induced TNF-alpha production in a THP-1 cell-based assay and against human p38 alpha MAP kinase in an isolated enzyme assay. The X-ray crystal structure of a bicyclic pyrazole inhibitor co-crystallized with mutated p38 (mp38) is presented.
Assuntos
Inibidores Enzimáticos/síntese química , Pirazóis/síntese química , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Humanos , Lipopolissacarídeos/farmacologia , Pirazóis/farmacologia , Relação Estrutura-Atividade , Fator de Necrose Tumoral alfa/biossíntese , Proteínas Quinases p38 Ativadas por Mitógeno/químicaRESUMO
Novel substituted [5,5]-bicyclic pyrzazolones are presented as inhibitors of tumor necrosis factor-alpha (TNF-alpha) production. Many of these compounds show low nanomolar activity against lipopolysaccaride (LPS)-induced TNF-alpha production in THP-1 cells. This class of molecules was co-crystallized with mutated p38, and several analogs showed good oral bioavailability in the rat. Oral activity of these compounds in the rat iodoacetate model for osteoarthritis is discussed.
Assuntos
Pirazolonas/química , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Administração Oral , Animais , Disponibilidade Biológica , Lipopolissacarídeos/farmacologia , Modelos Moleculares , Pirazolonas/administração & dosagem , Pirazolonas/farmacocinética , Pirazolonas/farmacologia , Ratos , Relação Estrutura-Atividade , Fator de Necrose Tumoral alfa/biossínteseRESUMO
2-Aryl-3-pyrimidinyl based tumor necrosis factor-alpha (TNF-alpha) inhibitors, which contain a novel bicyclic pyrazolone core, are described. Many showed low-nanomolar activity against lipopolysaccharide-induced TNF-alpha production in monocytic cells. Secondary screening data are presented for the pyrimidinyl bicyclic pyrazolones. Several of these analogues showed good oral bioavailability in rat and efficacy in the rat iodoacetate in vivo model.
Assuntos
Pirazóis/síntese química , Pirimidinas/síntese química , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Administração Oral , Animais , Disponibilidade Biológica , Humanos , Técnicas In Vitro , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Fosfotransferases/antagonistas & inibidores , Fosfotransferases/metabolismo , Pirazóis/química , Pirazóis/farmacologia , Pirimidinas/química , Pirimidinas/farmacologia , Ratos , Relação Estrutura-Atividade , Fator de Necrose Tumoral alfa/biossínteseRESUMO
4-Aryl-5-pyridyl and 4-aryl-5-pyrimidyl based inhibitors of TNF-alpha production, which contain a novel triazole 5-member heterocyclic core, are described. Many pyridyl triazoles containing either an alkyl ether or a substituted aryl side chain on the triazole core showed sub-micromolar activity against LPS-induced TNF-alpha, while pyrimidyl triazoles containing an ethoxymethyl side chain exhibited even better inhibitory activity. Secondary screening data are presented for the pyrimidyl triazoles. Triazole 14e combined excellent potency with good oral bioavailability in the rat.