RESUMO
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 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
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 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
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 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
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 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
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
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
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
Currently, there is no therapy for men with androgen-refractory prostate cancer that substantially extends survival. This report characterizes by in vitro and in vivo techniques a new chemotherapeutic that is composed of desacetyl-vinblastine covalently linked to a peptide that contains a peptide bond that can be hydrolyzed by prostate-specific antigen (PSA). This compound (referred to as vinblastine-conjugate) is minimally toxic to cells in culture which do not express PSA. In the presence of PSA, the peptide moiety is hydrolyzed, generating several highly toxic metabolites that contain vinblastine. Animals bearing PSA-positive human prostate tumors that were treated with the vinblastine-conjugate experienced a >99% reduction in PSA serum level. In contrast, animals bearing PSA-positive human prostate tumors treated with the cytotoxic metabolites derived from the PSA hydrolysis of the vinblastine-conjugate showed a nonsignificant change in both PSA and tumor weight values. The cell killing activity of the vinblastine-conjugate is PSA dependent because animals bearing non-PSA-producing human tumor xenografts had a nonsignificant increase in tumor weight after vinblastine-conjugate treatment. Exploratory efficacy/toxicity studies in LNCaP tumor-bearing nude mice were conducted with animals treated for 5 consecutive days with various doses of either the vinblastine-conjugate or a PSA-generated toxic metabolite (desacetyl-vinblastine). The desacetyl-vinblastine treatment resulted in 10-70% mortality with a very slight effect on tumor growth. In contrast, vinblastine-conjugate treatments resulted in no mortality, good to excellent antitumor efficacy, very slight to slight peripheral neuropathy and myelopathy, and slight to severe testicular degeneration. Similar treatment of beagle dogs with the vinblastine-conjugate showed even less toxicity. These data support the use of the PSA-hydrolyzable vinblastine-conjugate as an experimental therapy for prostate cancer in man.