RESUMO
A series of N-heterocyclic pyridinone catechol-O-methyltransferase (COMT) inhibitors were synthesized. Physicochemical properties, including ligand lipophilic efficiency (LLE) and clogP, were used to guide compound design and attempt to improve inhibitor pharmacokinetics. Incorporation of heterocyclic central rings provided improvements in physicochemical parameters but did not significantly reduce in vitro or in vivo clearance. Nevertheless, compound 11 was identified as a potent inhibitor with sufficient in vivo exposure to significantly affect the dopamine metabolites homovanillic acid (HVA) and dihydroxyphenylacetic acid (DOPAC), and indicate central COMT inhibition.
Assuntos
Inibidores de Catecol O-Metiltransferase/farmacologia , Catecol O-Metiltransferase/metabolismo , Compostos Heterocíclicos/farmacologia , Piridonas/farmacologia , Animais , Inibidores de Catecol O-Metiltransferase/síntese química , Inibidores de Catecol O-Metiltransferase/química , Relação Dose-Resposta a Droga , Compostos Heterocíclicos/síntese química , Compostos Heterocíclicos/química , Humanos , Modelos Moleculares , Estrutura Molecular , Piridonas/síntese química , Piridonas/química , Ratos , Relação Estrutura-AtividadeRESUMO
A series of [1,2,4]triazolo[3,4-f][1,6]naphthyridine allosteric dual inhibitors of Akt1 and 2 have been developed. These compounds have been shown to have potent dual Akt1 and 2 cell potency. The representative compound 13 provided potent inhibitory activity against Akt1 and 2 in vivo in a mouse model.
Assuntos
Química Farmacêutica/métodos , Naftiridinas/química , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Triazóis/química , Sítio Alostérico , Animais , Relação Dose-Resposta a Droga , Desenho de Fármacos , Canal de Potássio ERG1 , Canais de Potássio Éter-A-Go-Go/metabolismo , Concentração Inibidora 50 , Pulmão/efeitos dos fármacos , Camundongos , Modelos Químicos , Proteínas Proto-Oncogênicas c-akt/química , Relação Estrutura-AtividadeRESUMO
This letter describes the development of potent, allosteric dual Akt1 and Akt2 inhibitors with improved aqueous solubility (approximately 18 mg/mL) that translates into enhanced cell activity and caspase-3 induction.
Assuntos
Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Quinoxalinas/química , Quinoxalinas/farmacologia , Inibidores de Serina Proteinase/química , Inibidores de Serina Proteinase/farmacologia , Caspase 3/metabolismo , Linhagem Celular Tumoral , Ativação Enzimática , Células HT29 , Humanos , Cinética , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismo , Quinoxalinas/síntese química , Inibidores de Serina Proteinase/síntese química , Solubilidade , Água/químicaRESUMO
This letter shows inhibitor SAR on a pyridine series of allosteric Akt inhibitors to optimize enzymatic and cellular potency. We have optimized 2,3,5-trisubstituted pyridines to give potent Akt1 and Akt2 inhibitors in both enzyme and cell based assays. In addition, we will also highlight the pharmacokinetic profile of an optimized inhibitor that has low clearance and long half-life in dogs.
Assuntos
Sítio Alostérico , Neoplasias da Próstata/tratamento farmacológico , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Piridinas/química , Animais , Apoptose/efeitos dos fármacos , Caspases/metabolismo , Cães , Humanos , Masculino , Taxa de Depuração Metabólica , Estrutura Molecular , Neoplasias da Próstata/metabolismo , Inibidores de Proteínas Quinases/síntese química , Proteínas Proto-Oncogênicas c-akt/metabolismo , Relação Estrutura-Atividade , Ligante Indutor de Apoptose Relacionado a TNF/farmacologia , Células Tumorais CultivadasRESUMO
This paper describes the improvement of cell potency in a class of allosteric Akt 1 and 2 inhibitors. Key discoveries include identifying the solvent exposed region of the molecule and appending basic amines to enhance the physiochemical properties of the molecules. Findings from the structure-activity relationships are discussed.
Assuntos
Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Sítio Alostérico , Química Farmacêutica/métodos , Físico-Química/métodos , Desenho de Fármacos , Humanos , Concentração Inibidora 50 , Modelos Químicos , Fosforilação , Piperazinas/química , Inibidores de Proteínas Quinases/química , Proteínas Proto-Oncogênicas c-akt/química , Solventes/química , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
This letter details the attenuation of hERG in a class of Akt inhibitors through heteroatom insertions into aromatic rings. The development of a cell-active dual Akt 1 and 2 inhibitors devoid of hERG activity is discussed using structure-activity relationships.
Assuntos
Inibidores Enzimáticos/farmacologia , Canais de Potássio Éter-A-Go-Go/química , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Regulação Alostérica , Sítio Alostérico , Química Farmacêutica/métodos , Desenho de Fármacos , Canal de Potássio ERG1 , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Canais de Potássio Éter-A-Go-Go/metabolismo , Humanos , Concentração Inibidora 50 , Modelos Químicos , Relação Estrutura-AtividadeRESUMO
This communication reports a new synthetic route of pyridopyrimidines to facilitate their structural optimization in a library fashion and describes the development of pyridopyrimidines that have excellent enzymatic and cell potency against Akt1 and Akt2. This series also shows a high level of selectivity over other closely related kinases and significantly improved caspase-3 activity with the more optimized compounds.
Assuntos
Inibidores de Proteínas Quinases/síntese química , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Pirimidinas/síntese química , Pirimidinas/farmacologia , Aminas/síntese química , Aminas/química , Aminas/farmacologia , Caspase 3/metabolismo , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Humanos , Concentração Inibidora 50 , Piperidinas/síntese química , Piperidinas/química , Piperidinas/farmacologia , Inibidores de Proteínas Quinases/química , Pirimidinas/química , Relação Estrutura-Atividade , Ligante Indutor de Apoptose Relacionado a TNF/farmacologiaRESUMO
A series of naphthyridine and naphthyridinone allosteric dual inhibitors of Akt1 and 2 have been developed. These compounds have been optimized to have potent dual activity against the activated kinase as well as the activation of Akt in cells. One molecule in particular, compound 17, has potent inhibitory activity against Akt1 and 2 in vivo in a mouse lung and efficacy in a tumor xenograft model.
Assuntos
Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Naftiridinas/síntese química , Naftiridinas/farmacologia , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Animais , Antineoplásicos/química , Técnicas de Química Combinatória , Modelos Animais de Doenças , Desenho de Fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Camundongos , Naftiridinas/química , Proteínas Proto-Oncogênicas c-akt/metabolismo , Relação Estrutura-AtividadeRESUMO
This paper describes the rapid assembly of four different classes of potent Akt inhibitors from a common intermediate. Among them, a pyridopyrimidine series displayed the best intrinsic and cell potency against Akt1 and Akt2. This series also showed a promising pharmacokinetic profile and excellent selectivity over other closely related kinases.
Assuntos
Sítio Alostérico , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias da Próstata/tratamento farmacológico , Inibidores de Proteínas Quinases/síntese química , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Piridinas/química , Pirimidinas/química , Animais , Apoptose/efeitos dos fármacos , Caspases/metabolismo , Cães , Feminino , Humanos , Masculino , Taxa de Depuração Metabólica , Estrutura Molecular , Neoplasias Ovarianas/metabolismo , Fosforilação/efeitos dos fármacos , Neoplasias da Próstata/metabolismo , Inibidores de Proteínas Quinases/farmacocinética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Relação Estrutura-Atividade , Ligante Indutor de Apoptose Relacionado a TNF/farmacologia , Células Tumorais CultivadasRESUMO
We developed a high-throughput HTRF (homogeneous time-resolved fluorescence) assay for Akt kinase activity and screened approx. 270000 compounds for their ability to inhibit the three isoforms of Akt. Two Akt inhibitors were identified that exhibited isoenzyme specificity. The first compound (Akt-I-1) inhibited only Akt1 (IC50 4.6 microM) while the second compound (Akt-I-1,2) inhibited both Akt1 and Akt2 with IC50 values of 2.7 and 21 microM respectively. Neither compound inhibited Akt3 nor mutants lacking the PH (pleckstrin homology) domain at concentrations up to 250 microM. These compounds were reversible inhibitors, and exhibited a linear mixed-type inhibition against ATP and peptide substrate. In addition to inhibiting kinase activity of individual Akt isoforms, both inhibitors blocked the phosphorylation and activation of the corresponding Akt isoforms by PDK1 (phosphoinositide-dependent kinase 1). A model is proposed in which these inhibitors bind to a site formed only in the presence of the PH domain. Binding of the inhibitor is postulated to promote the formation of an inactive conformation. In support of this model, antibodies to the Akt PH domain or hinge region blocked the inhibition of Akt by Akt-I-1 and Akt-I-1,2. These inhibitors were found to be cell-active and to block phosphorylation of Akt at Thr308 and Ser473, reduce the levels of active Akt in cells, block the phosphorylation of known Akt substrates and promote TRAIL (tumour-necrosis-factor-related apoptosis-inducing ligand)-induced apoptosis in LNCap prostate cancer cells.
Assuntos
Proteínas Sanguíneas/química , Proteínas Sanguíneas/genética , Peptídeos/química , Peptídeos/genética , Fosfoproteínas/química , Fosfoproteínas/genética , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Proto-Oncogênicas/antagonistas & inibidores , Homologia de Sequência de Aminoácidos , Proteínas Quinases Dependentes de 3-Fosfoinositídeo , Trifosfato de Adenosina/metabolismo , Proteínas Reguladoras de Apoptose , Benzilaminas/farmacologia , Ligação Competitiva , Proteínas Sanguíneas/imunologia , Carcinoma/química , Carcinoma/metabolismo , Carcinoma/patologia , Caspases/metabolismo , Linhagem Celular Tumoral , Clonagem Molecular , Ativação Enzimática/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Feminino , Compostos Heterocíclicos com 2 Anéis/farmacologia , Humanos , Isoenzimas/antagonistas & inibidores , Isoenzimas/química , Masculino , Glicoproteínas de Membrana/farmacologia , Estrutura Molecular , Peptídeos/imunologia , Peptídeos/metabolismo , Fosfoproteínas/imunologia , Fosforilação/efeitos dos fármacos , Neoplasias da Próstata/química , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Proteínas Serina-Treonina Quinases/metabolismo , Estrutura Terciária de Proteína , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-akt , Quinoxalinas/farmacologia , Transdução de Sinais/fisiologia , Ligante Indutor de Apoptose Relacionado a TNF , Fator de Necrose Tumoral alfa/farmacologia , Neoplasias do Colo do Útero/química , Neoplasias do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/patologiaRESUMO
Recent studies indicate that dysregulation of the Akt/PKB family of serine/threonine kinases is a prominent feature of many human cancers. The Akt/PKB family is composed of three members termed Akt1/PKBalpha, Akt2/PKBbeta, and Akt3/PKBgamma. It is currently not known to what extent there is functional overlap between these family members. We have recently identified small molecule inhibitors of Akt. These compounds have pleckstrin homology domain-dependent, isozyme-specific activity. In this report, we present data showing the relative contribution that inhibition of the different isozymes has on the apoptotic response of tumor cells to a variety of chemotherapies. In multiple cell backgrounds, maximal induction of caspase-3 activity is achieved when both Akt1 and Akt2 are inhibited. This induction is not reversed by overexpression of functionally active Akt3. The level of caspase-3 activation achieved under these conditions is equivalent to that observed with the phosphatidylinositol-3-kinase inhibitor LY294002. We also show that in different tumor cell backgrounds inhibition of mammalian target of rapamycin, a downstream substrate of Akt, is less effective in inducing caspase-3 activity than inhibition of Akt1 and Akt2. This shows that the survival phenotype conferred by Akt can be mediated by signaling pathways independent of mammalian target of rapamycin in some tumor cell backgrounds. Finally, we show that inhibition of both Akt1 and Akt2 selectively sensitizes tumor cells, but not normal cells, to apoptotic stimuli.
Assuntos
Apoptose/fisiologia , Neoplasias/enzimologia , Inibidores de Proteínas Quinases/química , Inibidores de Proteínas Quinases/farmacologia , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Proto-Oncogênicas/antagonistas & inibidores , Proteínas Quinases Dependentes de 3-Fosfoinositídeo , Antibióticos Antineoplásicos/farmacologia , Caspase 3 , Caspases/metabolismo , Cromonas/farmacologia , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Ativação Enzimática , Humanos , Morfolinas/farmacologia , Isoformas de Proteínas/química , Isoformas de Proteínas/farmacologia , Proteínas Serina-Treonina Quinases/fisiologia , Proteínas Proto-Oncogênicas/fisiologia , Proteínas Proto-Oncogênicas c-akt , Sirolimo/farmacologia , Células Tumorais CultivadasRESUMO
3-Hydroxy-4-pyridinones and 5-hydroxy-4-pyrimidinones were identified as inhibitors of catechol-O-methyltransferase (COMT) in a high-throughput screen. These heterocyclic catechol mimics exhibit potent inhibition of the enzyme and an improved toxicity profile versus the marketed nitrocatechol inhibitors tolcapone and entacapone. Optimization of the series was aided by X-ray cocrystal structures of the novel inhibitors in complex with COMT and cofactors SAM and Mg(2+). The crystal structures suggest a mechanism of inhibition for these heterocyclic inhibitors distinct from previously disclosed COMT inhibitors.
RESUMO
A series of novel diaryl ether lactams have been identified as very potent dual inhibitors of protein farnesyltransferase (FTase) and protein geranylgeranyltransferase I (GGTase-I), enzymes involved in the prenylation of Ras. The structure of the complex formed between one of these compounds and FTase has been determined by X-ray crystallography. These compounds are the first reported to inhibit the prenylation of the important oncogene Ki-Ras4B in vivo. Unfortunately, doses sufficient to achieve this endpoint were rapidly lethal.
Assuntos
Alquil e Aril Transferases/antagonistas & inibidores , Antineoplásicos/síntese química , Animais , Antineoplásicos/química , Antineoplásicos/farmacologia , Proteínas de Transporte/metabolismo , Cristalografia por Raios X , Ensaios de Seleção de Medicamentos Antitumorais , Proteínas de Choque Térmico HSP40 , Proteínas de Choque Térmico/metabolismo , Humanos , Camundongos , Camundongos Nus , Modelos Moleculares , Transplante de Neoplasias , Neoplasias Experimentais/tratamento farmacológico , Prenilação de Proteína , Relação Estrutura-Atividade , Transplante Heterólogo , Células Tumorais Cultivadas , Proteínas rap1 de Ligação ao GTP/metabolismo , Proteínas ras/metabolismoRESUMO
Reduced dopamine neurotransmission in the prefrontal cortex has been implicated as causal for the negative symptoms and cognitive deficit associated with schizophrenia; thus, a compound which selectively enhances dopamine neurotransmission in the prefrontal cortex may have therapeutic potential. Inhibition of catechol-O-methyltransferase (COMT, EC 2.1.1.6) offers a unique advantage, since this enzyme is the primary mechanism for the elimination of dopamine in cortical areas. Since membrane bound COMT (MB-COMT) is the predominant isoform in human brain, a high throughput screen (HTS) to identify novel MB-COMT specific inhibitors was completed. Subsequent optimization led to the identification of novel, non-nitrocatechol COMT inhibitors, some of which interact specifically with MB-COMT. Compounds were characterized for in vitro efficacy versus human and rat MB and soluble (S)-COMT. Select compounds were administered to male Wistar rats, and ex vivo COMT activity, compound levels in plasma and cerebrospinal fluid (CSF), and CSF dopamine metabolite levels were determined as measures of preclinical efficacy. Finally, novel non-nitrocatechol COMT inhibitors displayed less potent uncoupling of the mitochondrial membrane potential (MMP) compared to tolcapone as well as nonhepatotoxic entacapone, thus mitigating the risk of hepatotoxicity.
Assuntos
Antipsicóticos/farmacocinética , Inibidores de Catecol O-Metiltransferase , Catecol O-Metiltransferase/metabolismo , Inibidores Enzimáticos/farmacologia , Animais , Antipsicóticos/síntese química , Benzofenonas/química , Benzofenonas/farmacologia , Biomarcadores , Western Blotting , Catecol O-Metiltransferase/isolamento & purificação , Membrana Celular/enzimologia , Membrana Celular/metabolismo , Dopamina/metabolismo , Inibidores Enzimáticos/química , Humanos , Isoenzimas/antagonistas & inibidores , Isoenzimas/isolamento & purificação , Isoenzimas/metabolismo , Masculino , Metaloproteinases da Matriz/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Nitrofenóis/química , Nitrofenóis/farmacologia , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Proteínas Recombinantes/química , Esquizofrenia/tratamento farmacológico , Especificidade por Substrato , TolcaponaRESUMO
This letter describes the discovery of a novel series of dual Akt1/Akt2 kinase inhibitors, based on a 2,3,5-trisubstituted pyridine scaffold. Compounds from this series, which contain a 5-tetrazolyl moiety, exhibit more potent inhibition of Akt2 than Akt1.
Assuntos
Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Proto-Oncogênicas/antagonistas & inibidores , Piridinas/química , Piridinas/farmacologia , Animais , Permeabilidade da Membrana Celular , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacocinética , Inibidores Enzimáticos/farmacologia , Humanos , Concentração Inibidora 50 , Proteínas Proto-Oncogênicas c-akt , Piridinas/farmacocinética , Relação Estrutura-AtividadeRESUMO
This letter describes the development of two series of potent and selective allosteric Akt kinase inhibitors that display an unprecedented level of selectivity for either Akt1, Akt2 or both Akt1/Akt2. An iterative analog library synthesis approach quickly provided a highly selective Akt1/Akt2 inhibitor that induces apoptosis in tumor cells and inhibits Akt phosphorylation in vivo.
Assuntos
Regulação Alostérica , Inibidores Enzimáticos/síntese química , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Proto-Oncogênicas/antagonistas & inibidores , Apoptose/efeitos dos fármacos , Humanos , Concentração Inibidora 50 , Isoenzimas/antagonistas & inibidores , Fosforilação/efeitos dos fármacos , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-akt , Quinoxalinas/síntese química , Quinoxalinas/farmacologia , Relação Estrutura-AtividadeRESUMO
Compound 1 has been shown to be a dual prenylation inhibitor with FPTase (IC50=2 nM) and GGPTase-I (IC50=95 nM). Analogues of 1, which replaced the cyanophenyl group with various biaryls, led to the discovery of highly potent dual FPTase/GGPTase-I inhibitors. 4-trifluoromethylphenyl, trifluoropentynyl, and trifluoropentyl were identified as good p-cyano replacements.
Assuntos
Alquil e Aril Transferases/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/químicaRESUMO
A series of macrocyclic piperazinone compounds with dual farnesyltransferase/geranylgeranyltransferase-I inhibitory activity was prepared. These compounds were found to be potent inhibitors of protein prenylation in cell culture. A hypothesis for the binding mode of compound 3o in FPTase is proposed.
Assuntos
Alquil e Aril Transferases/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Compostos Heterocíclicos/farmacologia , Piperazinas/farmacologia , Células Tumorais Cultivadas/efeitos dos fármacos , Inibidores Enzimáticos/química , Compostos Heterocíclicos/química , Humanos , Concentração Inibidora 50 , Estrutura Molecular , Piperazinas/química , Prenilação de Proteína , Relação Estrutura-Atividade , Células Tumorais Cultivadas/enzimologiaRESUMO
We have prepared a series of potent, dual inhibitors of the prenyl transferases farnesyl protein transferase (FPTase) and geranyl-geranyl protein transferase I (GGPTase). The compounds were shown to possess potent activity against both enzymes in cell culture. Mechanistic analysis has shown that the compounds are CAAX competitive for FPTase inhibition but geranyl-geranyl pyrophosphate (GGPP) competitive for GGPTase inhibiton.