RESUMO
Smoothened (Smo) antagonists are emerging as new therapies for the treatment of neoplasias with aberrantly reactivated hedgehog (Hh) signaling pathway. A novel series of 4-[3-(quinolin-2-yl)-1,2,4-oxadiazol-5-yl]piperazinyl ureas as smoothened antagonists was recently described, herein the series has been further optimized through the incorporation of a basic amine into the urea. This development resulted in identification of some exceptionally potent smoothened antagonists with low serum shifts, however, reductive ring opening on the 1,2,4-oxadiazole in rats limits the applicability of these compounds in in vivo studies.
Assuntos
Amidas/farmacologia , Piperazinas/farmacologia , Receptores Acoplados a Proteínas G/antagonistas & inibidores , Amidas/síntese química , Amidas/química , Estrutura Molecular , Piperazinas/síntese química , Piperazinas/química , Receptor Smoothened , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
We describe an extensive SAR study in the 6-[4-fluoro-3-(substituted)benzyl]-4,5-dimethylpyridazin-3(2H)-one series which led to the identification of potent PARP-1 inhibitors, capable of inhibiting the proliferation of BRCA-1 deficient cancer cells in the low nanomolar range, and displaying >100-fold selectivity over the BRCA wild type counterparts. The series of compounds was devoid of hERG channel activity, and CYP inhibition and induction liabilities. Several analogs were stable in rat and human liver microsomes and displayed moderate rat clearance, with urinary excretion of parent as the major route of elimination.
Assuntos
Proteína BRCA1/deficiência , Piperazinas/síntese química , Inibidores de Poli(ADP-Ribose) Polimerases , Piridazinas/síntese química , Animais , Proteína BRCA1/genética , Células HeLa , Humanos , Microssomos Hepáticos/efeitos dos fármacos , Microssomos Hepáticos/enzimologia , Piperazinas/metabolismo , Piperazinas/farmacologia , Poli(ADP-Ribose) Polimerase-1 , Poli(ADP-Ribose) Polimerases/metabolismo , Piridazinas/metabolismo , Piridazinas/farmacologia , RatosRESUMO
Herein we describe the discovery of a novel series of pyrrolo[1,2-a]pyrazin-1(2H)-one PARP inhibitors. Optimization led to compounds that display excellent PARP-1 enzyme potency and inhibit the proliferation of BRCA deficient cells in the low double-digit nanomolar range showing excellent selectivity over BRCA proficient cancer cells.
Assuntos
Inibidores de Poli(ADP-Ribose) Polimerases , Pirazinas/química , Pirazinas/metabolismo , Animais , Proteína BRCA1/deficiência , Proteína BRCA1/metabolismo , Linhagem Celular Tumoral , Cristalografia por Raios X , Células HeLa , Humanos , Indolizinas/química , Indolizinas/metabolismo , Microssomos Hepáticos/efeitos dos fármacos , Microssomos Hepáticos/enzimologia , Poli(ADP-Ribose) Polimerase-1 , Poli(ADP-Ribose) Polimerases/metabolismo , Ligação Proteica/fisiologia , Ratos , Relação Estrutura-AtividadeRESUMO
PARP inhibitors have been demonstrated to retard intracellular DNA repair and therefore sensitize tumor cells to cytotoxic agents or ionizing radiation. We report the identification of a novel class of PARP1 inhibitors, containing a pyrrolo moiety fused to a dihydroisoquinolinone, derived from virtual screening of the proprietary collection. SAR exploration around the nitrogen of the aminoethyl appendage chain of 1 led to compounds that displayed low nanomolar activity in a PARP1 enzymatic assay.
Assuntos
Inibidores de Poli(ADP-Ribose) Polimerases , Quinolonas/química , Antineoplásicos/farmacologia , Sítios de Ligação , Química Farmacêutica/métodos , Cristalografia por Raios X/métodos , Reparo do DNA , Desenho de Fármacos , Humanos , Ligação de Hidrogênio , Concentração Inibidora 50 , Ligantes , Modelos Químicos , Polímeros/química , Relação Estrutura-AtividadeRESUMO
In the context of HIV-integrase, dihydroxypyrimidine and N-methyl pyrimidone inhibitors the cellular activity of this class of compounds has been optimized by the introduction of a simple methyl substituent in the alpha-position of the C-2 side chains. Enhanced passive membrane permeability has been identified as the key factor driving the observed cell-based activity improvement. The rat PK profile of the alpha-methyl derivative 26a was also improved over its des-methyl exact analog.
Assuntos
Inibidores de Integrase de HIV/química , Integrase de HIV/química , Pirimidinas/química , Pirimidinonas/química , Animais , Permeabilidade da Membrana Celular , Integrase de HIV/metabolismo , Inibidores de Integrase de HIV/síntese química , Inibidores de Integrase de HIV/farmacocinética , Humanos , Ligação Proteica , Pirimidinas/síntese química , Pirimidinas/farmacocinética , Pirimidinonas/síntese química , Pirimidinonas/farmacocinética , RatosRESUMO
We report the synthesis and biological evaluation of N-[(1-aryl-1H-indazol-5-yl)methyl]amide derivatives as Smoothened antagonists and inhibitors of the Hedgehog pathway. Identification of the lead structure 1 by HTS, followed by SAR study on the amide and aryl portions led to the discovery of antagonists with nanomolar activity.
Assuntos
Amidas/síntese química , Indazóis/síntese química , Receptores Acoplados a Proteínas G/antagonistas & inibidores , Receptores Acoplados a Proteínas G/química , Amidas/farmacologia , Catálise , Linhagem Celular , Química Farmacêutica/métodos , Técnicas de Química Combinatória/métodos , Desenho de Fármacos , Avaliação Pré-Clínica de Medicamentos , Humanos , Indazóis/farmacologia , Concentração Inibidora 50 , Modelos Químicos , Transdução de Sinais , Receptor Smoothened , Relação Estrutura-AtividadeRESUMO
A novel series of pyrazolo[1,5-a]quinazolin-5(4H)-one derivatives proved to be a potent class of PARP-1 inhibitors. An extensive SAR around the 3-position of pyrazole in the scaffold led to the discovery of amides derivatives as low nanomolar PARP-1 inhibitors.
Assuntos
Inibidores Enzimáticos/síntese química , Inibidores de Poli(ADP-Ribose) Polimerases , Pirazóis/síntese química , Amidas/química , Química Orgânica/métodos , Química Farmacêutica/métodos , Desenho de Fármacos , Avaliação Pré-Clínica de Medicamentos , Inibidores Enzimáticos/farmacologia , Células HeLa , Humanos , Concentração Inibidora 50 , Modelos Químicos , Estrutura Molecular , Pirazóis/farmacologia , Quinazolinonas/síntese química , Quinazolinonas/farmacologia , Relação Estrutura-AtividadeRESUMO
HIV integrase is one of the three enzymes encoded by HIV genome and is essential for viral replication, but integrase inhibitors as marketed drugs have just very recently started to emerge. In this study, we show the evolution from the N-methylpyrimidinone structure to bicyclic pyrimidinones. Introduction of a suitably substituted amino moiety modulated the physical-chemical properties of the molecules and conferred nanomolar activity in the inhibition of spread of HIV-1 infection in cell culture. An extensive SAR study led to sulfamide (R)- 22b, which inhibited the strand transfer with an IC50 of 7 nM and HIV infection in MT4 cells with a CIC95 of 44 nM, and ketoamide (S)- 28c that inhibited strand transfer with an IC50 of 12 nM and the HIV infection in MT4 cells with a CIC95 of 13 nM and exhibited a good pharmacokinetic profile when dosed orally to preclinical species.
Assuntos
Aminopiridinas/síntese química , Azepinas/síntese química , Compostos Bicíclicos Heterocíclicos com Pontes/síntese química , Inibidores de Integrase de HIV/síntese química , Integrase de HIV/metabolismo , Pirimidinonas/síntese química , Administração Oral , Aminopiridinas/farmacocinética , Aminopiridinas/farmacologia , Animais , Azepinas/farmacocinética , Azepinas/farmacologia , Disponibilidade Biológica , Compostos Bicíclicos Heterocíclicos com Pontes/farmacocinética , Compostos Bicíclicos Heterocíclicos com Pontes/farmacologia , Linhagem Celular , Cães , Integrase de HIV/genética , Inibidores de Integrase de HIV/farmacocinética , Inibidores de Integrase de HIV/farmacologia , HIV-1/efeitos dos fármacos , Humanos , Macaca mulatta , Microssomos Hepáticos/metabolismo , Pirimidinonas/farmacocinética , Pirimidinonas/farmacologia , Ratos , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
Trifluoroacetylthiophene carboxamides have recently been reported to be class II HDAC inhibitors, with moderate selectivity. Exploration of replacements for the carboxamide with bioisosteric pentatomic heteroaromatic like 1,3,4-oxadiazoles, 1,2,4-oxadiazoles and 1,3-thiazoles, led to the discovery that 2-trifluoroacetylthiophene 1,3,4-oxadiazole derivatives are very potent low nanomolar HDAC4 inhibitors, highly selective over class I HDACs (HDAC 1 and 3), and moderately stable in HCT116 cell culture.
Assuntos
Inibidores de Histona Desacetilases , Oxidiazóis/síntese química , Oxidiazóis/farmacologia , Tiofenos/síntese química , Tiofenos/farmacologia , Técnicas de Química Combinatória , Desenho de Fármacos , Células HCT116 , Histona Acetiltransferases/antagonistas & inibidores , Histona Desacetilases/classificação , Humanos , Estrutura Molecular , Oxidiazóis/química , Relação Estrutura-Atividade , Tiofenos/químicaRESUMO
5-(Trifluoroacetyl)thiophene-2-carboxamides were found to be potent and selective class II HDAC inhibitors. This paper describes their further development and the investigation on the cause for the lack of cell-based activity. A rapid screening assay was set up which enabled the identification of more metabolic stable compounds as potent and selective class II HDAC inhibitors.
Assuntos
Amidas/síntese química , Amidas/farmacologia , Inibidores de Histona Desacetilases , Tiofenos/síntese química , Tiofenos/farmacologia , Amidas/química , Hidrocarboneto de Aril Hidroxilases/antagonistas & inibidores , Citocromo P-450 CYP2C9 , Desenho de Fármacos , Células HCT116 , Células HeLa , Hepatócitos/efeitos dos fármacos , Histona Desacetilases/classificação , Humanos , Microssomos Hepáticos/efeitos dos fármacos , Estrutura Molecular , Relação Estrutura-Atividade , Tiofenos/química , Tubulina (Proteína)/efeitos dos fármacosRESUMO
Human immunodeficiency virus type-1 (HIV-1) integrase, one of the three constitutive viral enzymes required for replication, is a rational target for chemotherapeutic intervention in the treatment of AIDS that has also recently been confirmed in the clinical setting. We report here on the design and synthesis of N-benzyl-5,6-dihydroxypyrimidine-4-carboxamides as a class of agents which exhibits potent inhibition of the HIV-integrase-catalyzed strand transfer process. In the current study, structural modifications on these molecules were made in order to examine effects on HIV-integrase inhibitory potencies. One of the most interesting compounds for this series is 2-[1-(dimethylamino)-1-methylethyl]-N-(4-fluorobenzyl)-5,6-dihydroxypyrimidine-4-carboxamide 38, with a CIC95 of 78 nM in the cell-based assay in the presence of serum proteins. The compound has favorable pharmacokinetic properties in preclinical species (rats, dogs, and monkeys) and shows no liabilities in several counterscreening assays, highlighting its potential as a clinically useful antiviral agent.
Assuntos
Inibidores de Integrase de HIV/síntese química , HIV-1/efeitos dos fármacos , Piridinas/síntese química , Pirimidinas/síntese química , Animais , Disponibilidade Biológica , Proteínas Sanguíneas/metabolismo , Linhagem Celular Tumoral , Cães , Inibidores de Integrase de HIV/farmacocinética , Inibidores de Integrase de HIV/farmacologia , Meia-Vida , Humanos , Macaca mulatta , Ligação Proteica , Piridinas/química , Piridinas/farmacologia , Pirimidinas/farmacocinética , Pirimidinas/farmacologia , Ratos , Relação Estrutura-Atividade , Replicação ViralRESUMO
The human immunodeficiency virus type-1 (HIV-1) encodes three enzymes essential for viral replication: a reverse transcriptase, a protease, and an integrase. The latter is responsible for the integration of the viral genome into the human genome and, therefore, represents an attractive target for chemotherapeutic intervention against AIDS. A drug based on this mechanism has not yet been approved. Benzyl-dihydroxypyrimidine-carboxamides were discovered in our laboratories as a novel and metabolically stable class of agents that exhibits potent inhibition of the HIV integrase strand transfer step. Further efforts led to very potent compounds based on the structurally related N-Me pyrimidone scaffold. One of the more interesting compounds in this series is the 2-N-Me-morpholino derivative 27a, which shows a CIC95 of 65 nM in the cell in the presence of serum. The compound has favorable pharmacokinetic properties in three preclinical species and shows no liabilities in several counterscreening assays.
Assuntos
Inibidores de Integrase de HIV/síntese química , Integrase de HIV/química , HIV-1/efeitos dos fármacos , Morfolinas/síntese química , Pirimidinonas/síntese química , Administração Oral , Animais , Disponibilidade Biológica , Proteínas Sanguíneas/metabolismo , Linhagem Celular Tumoral , Cães , Inibidores de Integrase de HIV/farmacocinética , Inibidores de Integrase de HIV/farmacologia , HIV-1/enzimologia , HIV-1/fisiologia , Humanos , Macaca mulatta , Morfolinas/farmacocinética , Morfolinas/farmacologia , Ligação Proteica , Pirimidinonas/farmacocinética , Pirimidinonas/farmacologia , Ratos , Estereoisomerismo , Relação Estrutura-Atividade , Replicação Viral/efeitos dos fármacosRESUMO
The dihydroxypyrimidine carboxamide 4a was discovered as a potent and selective HIV integrase strand transfer inhibitor. The optimization of physicochemical properties, pharmacokinetic profiles, and potency led to the identification of 13 in the dihydroxypyrimidine series and 18 in the N-methylpyrimidinone series having low nanomolar activity in the cellular HIV spread assay in the presence of 50% normal human serum and very good pharmacokinetics in preclinical species.
Assuntos
Amidas/síntese química , Inibidores de Integrase de HIV/síntese química , Pirimidinas/síntese química , Administração Oral , Amidas/química , Amidas/farmacologia , Animais , Disponibilidade Biológica , Cães , Inibidores de Integrase de HIV/farmacocinética , Inibidores de Integrase de HIV/farmacologia , HIV-1/efeitos dos fármacos , Humanos , Técnicas In Vitro , Macaca mulatta , Pirimidinas/química , Pirimidinas/farmacologia , Pirimidinonas/síntese química , Pirimidinonas/farmacocinética , Pirimidinonas/farmacologia , Ratos , Soro , Relação Estrutura-Atividade , Replicação ViralRESUMO
A strategy to obtain a fully orthogonal estrogen-receptor-based gene switch responsive to molecules with acceptable pharmacological properties is presented. From a series of tetrahydrofluorenones active on the wild-type estrogen receptor (ER) an inactive analogue is chosen as a new lead compound. Coevolution of receptor mutants and ligands leads to an ER-based gene switch suitable for studies in animal models.
Assuntos
Fluorenos/síntese química , Receptores de Estrogênio/efeitos dos fármacos , Sítios de Ligação , Estradiol/química , Receptor alfa de Estrogênio/efeitos dos fármacos , Receptor alfa de Estrogênio/genética , Receptor beta de Estrogênio/efeitos dos fármacos , Receptor beta de Estrogênio/genética , Fluorenos/química , Fluorenos/farmacologia , Células HeLa , Humanos , Ligantes , Modelos Moleculares , Estrutura Molecular , Mutação , Receptores de Estrogênio/genética , Relação Estrutura-AtividadeRESUMO
It is highly desirable to design ligand-dependent transcription regulation systems based on transactivators unresponsive to endogenous ligands but induced by synthetic small molecules unable to activate endogenous receptors. Using molecular modeling and yeast selection, we identified an estrogen receptor ligand binding domain double mutant (L384M, M421G) with decreased affinity to estradiol and enhanced binding to compounds inactive on estrogen receptors. Nonresponsiveness to estrogen was achieved by additionally adding the G521R substitution while introducing an "antagonistic-type" side chain in the compound, as in 4-hydroxytamoxifen. The triple-substituted ligand binding domain is insensitive to physiological concentrations of estradiol and has nanomolar affinity for the ligand. In this binary system, both receptor and ligand are, therefore, reciprocally specific. The mutated variant in the context of a chimeric transcription factor provides tight, ligand-dependent regulation of reporter gene expression.
Assuntos
Estradiol/análogos & derivados , Receptor alfa de Estrogênio/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Transcrição Gênica , Substituição de Aminoácidos , Sítios de Ligação/genética , Desenho de Fármacos , Estradiol/metabolismo , Estradiol/farmacologia , Receptor alfa de Estrogênio/genética , Genes Reporter , Humanos , Cinética , Ligantes , Proteínas Recombinantes de Fusão/genética , Fatores de Transcrição/genética , Leveduras/genéticaRESUMO
5-Aryl-2-(trifluoroacetyl)thiophenes were identified as a new series of class II HDAC inhibitors (HDACi). Further development of this new series led to compounds such as 6h, a potent inhibitor of HDAC4 and HDAC6 (HDAC4 WT IC(50) = 310 nM, HDAC6 IC(50) = 70 nM) that displays 40-fold selectivity over HDAC1 and improved stability in HCT116 cancer cells (t(1/2) = 11 h). Compounds 6h and 2 show inhibition of alpha-tubulin deacetylation in HCT116 cells at 1 microM concentration and antiproliferation effects only at concentrations where inhibition of histone H3 deacetylation is observed.
Assuntos
Antineoplásicos/síntese química , Histona Desacetilase 2/antagonistas & inibidores , Proteínas Repressoras/antagonistas & inibidores , Tiofenos/síntese química , Acetilação , Antineoplásicos/química , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Histona Desacetilase 1/antagonistas & inibidores , Desacetilase 6 de Histona , Histona Desacetilases/metabolismo , Histonas/metabolismo , Humanos , Isoenzimas/antagonistas & inibidores , Relação Estrutura-Atividade , Tiofenos/química , Tiofenos/farmacologia , Tubulina (Proteína)/metabolismoRESUMO
Human immunodeficiency virus type-1 (HIV-1) integrase is one of the three virally encoded enzymes required for replication and therefore a rational target for chemotherapeutic intervention in the treatment of HIV-1 infection. We report here the discovery of Raltegravir, the first HIV-integrase inhibitor approved by FDA for the treatment of HIV infection. It derives from the evolution of 5,6-dihydroxypyrimidine-4-carboxamides and N-methyl-4-hydroxypyrimidinone-carboxamides, which exhibited potent inhibition of the HIV-integrase catalyzed strand transfer process. Structural modifications on these molecules were made in order to maximize potency as HIV-integrase inhibitors against the wild type virus, a selection of mutants, and optimize the selectivity, pharmacokinetic, and metabolic profiles in preclinical species. The good profile of Raltegravir has enabled its progression toward the end of phase III clinical trials for the treatment of HIV-1 infection and culminated with the FDA approval as the first HIV-integrase inhibitor for the treatment of HIV-1 infection.
Assuntos
Infecções por HIV/tratamento farmacológico , Inibidores de Integrase de HIV/farmacologia , Pirrolidinonas/farmacologia , Administração Oral , Área Sob a Curva , Disponibilidade Biológica , Inibidores de Integrase de HIV/administração & dosagem , Inibidores de Integrase de HIV/farmacocinética , Inibidores de Integrase de HIV/uso terapêutico , Meia-Vida , Humanos , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Pirrolidinonas/administração & dosagem , Pirrolidinonas/farmacocinética , Pirrolidinonas/uso terapêutico , Raltegravir PotássicoRESUMO
LasR regulates toxin production in Pseudomonas aeruginosa and its inhibition can attenuate the virulence of this opportunistic human pathogen. To aid studies of interactions with inhibitors, we report the NMR backbone assignments for the dimeric LasR ligand-binding domain.
Assuntos
Proteínas de Bactérias/química , Espectroscopia de Ressonância Magnética/métodos , Pseudomonas aeruginosa/metabolismo , Transativadores/química , Sequência de Aminoácidos , Isótopos de Carbono/química , Peso Molecular , Isótopos de Nitrogênio/química , Estrutura Terciária de Proteína , PrótonsRESUMO
Many Gram-negative bacteria communicate via molecules called autoinducers to coordinate the activities of their populations. Such communication is termed quorum sensing and can regulate pathogenic virulence factor production and antimicrobial resistance. The quorum sensing system of Pseudomonas aeruginosa is currently the most intensively researched, because this bacterium is an opportunistic human pathogen annually responsible for the death of thousands of cystic fibrosis sufferers and many other immunocompromised individuals. Quorum sensing inhibitors can attenuate the pathogenicity of P. aeruginosa. Here we present the crystal structure of the P. aeruginosa LasR ligand-binding domain bound to its autoinducer 3-oxo-C(12)-acylhomoserine lactone. The structure is a symmetrical dimer, with each monomer exhibiting an alpha-beta-alpha fold similar to the TraR and SdiA quorum sensing proteins of Agrobacterium tumefaciens and Escherichia coli. The structure was determined up to 1.8-A resolution and reveals the atomic interactions between LasR and its autoinducer. The monomer structures of LasR, TraR, and SdiA are comparable but display differences in their quaternary organization. Inspection of their binding sites shows some unexpected variations resulting in quite different conformations of their bound autoinducers. We modeled interactions between LasR and various quorum sensing inhibitors, yielding insight into their possible mechanisms of action. The structure also provides a platform for the optimization, or de novo design, of quorum sensing inhibitors.
Assuntos
4-Butirolactona/análogos & derivados , Proteínas de Bactérias/química , Proteínas de Ligação a DNA/química , Lactonas/química , Pseudomonas aeruginosa/química , Percepção de Quorum , Transativadores/química , Fatores de Virulência/química , 4-Butirolactona/química , 4-Butirolactona/metabolismo , Proteínas de Bactérias/metabolismo , Cristalografia por Raios X , Fibrose Cística/complicações , Fibrose Cística/metabolismo , Fibrose Cística/mortalidade , Proteínas de Ligação a DNA/metabolismo , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/metabolismo , Humanos , Infecções Oportunistas/etiologia , Infecções Oportunistas/metabolismo , Infecções Oportunistas/mortalidade , Estrutura Terciária de Proteína , Infecções por Pseudomonas/etiologia , Infecções por Pseudomonas/metabolismo , Infecções por Pseudomonas/mortalidade , Pseudomonas aeruginosa/metabolismo , Pseudomonas aeruginosa/patogenicidade , Percepção de Quorum/fisiologia , Homologia Estrutural de Proteína , Transativadores/metabolismo , Fatores de Virulência/metabolismoRESUMO
A series of aryltetrazolylacetanilides was synthesized and evaluated as HIV-1 non-nucleoside reverse transcriptase inhibitors on wild-type virus and on the clinically relevant K103N mutant strain. Extensive SAR investigation led to potent compounds, with nanomolar activity on K103N, and orally bioavailable in rats.