RESUMO
(2'R)-Ethynyl uridine 3, and its (2'S)-diastereomer 10, are synthesised in a divergent fashion from the inexpensive parent nucleoside. Both nucleoside analogues are obtained from a total of 5 simple synthetic steps and 3 trivial column chromatography purifications. To evaluate their effectiveness against HCV NS5B polymerase, the nucleosides were converted to their respective 5'-O-triphosphates. Subsequently, this lead to the discovery of the 2'-ß-ethynyl 18 and -propynyl 20 nucleotides having significantly improved potency over Sofosbuvir triphosphate 24.
Assuntos
Antivirais/farmacologia , Hepacivirus/efeitos dos fármacos , Nucleosídeos/farmacologia , Uridina/farmacologia , Proteínas não Estruturais Virais/antagonistas & inibidores , Antivirais/síntese química , Antivirais/química , Relação Dose-Resposta a Droga , Testes de Sensibilidade Microbiana , Conformação Molecular , Nucleosídeos/síntese química , Nucleosídeos/química , Relação Estrutura-Atividade , Uridina/análogos & derivados , Uridina/químicaRESUMO
The discovery of lead compound 2e was described. Its covalent binding to HCV NS5B polymerase enzyme was investigated by X-ray analysis. The results of distribution, metabolism and pharmacokinetics were reported. Compound 2e was demonstrated to be potent (replicon GT-1b EC50 = 0.003 µM), highly selective, and safe in in vitro and in vivo assays.
Assuntos
Inibidores Enzimáticos/química , Hepacivirus/enzimologia , Indóis/química , Quinolinas/química , Proteínas não Estruturais Virais/antagonistas & inibidores , Animais , Cristalografia por Raios X , Cães , Avaliação Pré-Clínica de Medicamentos , Ativação Enzimática/efeitos dos fármacos , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/farmacocinética , Inibidores Enzimáticos/farmacologia , Haplorrinos , Humanos , Indóis/síntese química , Indóis/farmacocinética , Indóis/farmacologia , Masculino , Simulação de Dinâmica Molecular , Ligação Proteica , Estrutura Terciária de Proteína , Quinolinas/síntese química , Quinolinas/farmacocinética , Quinolinas/farmacologia , Ratos , Ratos Sprague-Dawley , Proteínas não Estruturais Virais/metabolismoRESUMO
Development of SAR at the C2 position of indole lead 1, a palm site inhibitor of HCV NS5B polymerase (NS5B IC(50)=0.053µM, replicon EC(50)=4.8µM), is described. Initial screening identified an acyl sulfonamide moiety as an isostere for the C2 carboxylic acid group. Further SAR investigation resulted in identification of acyl sufonamide analog 7q (NS5B IC(50)=0.039µM, replicon EC(50)=0.011µM) with >100-fold improved replicon activity.
Assuntos
Antivirais/farmacologia , Indóis/química , Proteínas não Estruturais Virais/antagonistas & inibidores , Química Farmacêutica/métodos , Cristalografia por Raios X/métodos , Desenho de Fármacos , Humanos , Ligação de Hidrogênio , Concentração Inibidora 50 , Modelos Químicos , Modelos Moleculares , Conformação Molecular , Relação Estrutura-Atividade , Sulfonamidas/químicaRESUMO
SAR development of indole-based palm site inhibitors of HCV NS5B polymerase exemplified by initial indole lead 1 (NS5B IC(50)=0.9 µM, replicon EC(50)>100 µM) is described. Structure-based drug design led to the incorporation of novel heterocyclic moieties at the indole C3-position which formed a bidentate interaction with the protein backbone. SAR development resulted in leads 7q (NS5B IC(50)=0.032 µM, replicon EC(50)=1.4 µM) and 7r (NS5B IC(50)=0.017 µM, replicon EC(50)=0.3 µM) with improved enzyme and replicon activity.
Assuntos
Descoberta de Drogas , Inibidores Enzimáticos/farmacologia , Compostos Heterocíclicos/farmacologia , Indóis/farmacologia , Proteínas não Estruturais Virais/antagonistas & inibidores , Ácidos Carboxílicos , Domínio Catalítico/efeitos dos fármacos , Cristalografia por Raios X , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Compostos Heterocíclicos/síntese química , Compostos Heterocíclicos/química , Indóis/síntese química , Indóis/química , Modelos Moleculares , Estrutura Molecular , Estereoisomerismo , Relação Estrutura-Atividade , Proteínas não Estruturais Virais/metabolismoRESUMO
Several analogs of aristolochic acids were isolated and derivatized into their lactam derivatives to study their inhibition in CDK2 assay. The study helped to derive some conclusions about the structure-activity relation around the phenanthrin moiety. Semi-synthetic aristolactam 21 showed good activity with inhibition IC50 of 35 nM in CDK2 assay. The activity of this compound was comparable to some of the most potent synthetic compounds reported in the literature.
Assuntos
Ácidos Aristolóquicos/síntese química , Proteína Quinase CDC2/antagonistas & inibidores , Pirazóis/síntese química , Quinolinas/síntese química , Ácidos Aristolóquicos/química , Ácidos Aristolóquicos/farmacologia , Linhagem Celular Tumoral , Simulação por Computador , Cristalografia por Raios X , Humanos , Concentração Inibidora 50 , Modelos Moleculares , Estrutura Molecular , Pirazóis/química , Pirazóis/farmacologia , Quinolinas/química , Quinolinas/farmacologia , Relação Estrutura-AtividadeRESUMO
Investigation of unexpected levels of impurities in Intron product has revealed the presence of low levels of impurities leached from the silicone tubing (Rehau RAU-SIK) on the Bosch filling line. In order to investigate the effect of these compounds (1a, 1b and 2) on humans, they were isolated identified and synthesized. They were extracted from the tubing by stirring in Intron placebo at room temperature for 72 h and were enriched on a reverse phase CHP-20P column, eluting with gradient aqueous ACN and were separated by HPLC. Structural elucidation of 1a, 1b and 2 by MS and NMR studies demonstrated them to be halogenated biphenyl carboxylic acids. The structures were confirmed by independent synthesis. Levels of extractable impurities in first filled vials of actual production are estimated to be in the range of 0.01-0.55 microg/vial for each leached impurity. Potential toxicity of these extractables does not represent a risk for patients under the conditions of clinical use.
Assuntos
Contaminação de Medicamentos , Interferon-alfa/química , Interferon-alfa/isolamento & purificação , Compostos Orgânicos/química , Silicones/química , Soluções Tampão , Química Farmacêutica , Cromatografia Líquida de Alta Pressão/métodos , Liofilização , Humanos , Concentração de Íons de Hidrogênio , Interferon alfa-2 , Espectroscopia de Ressonância Magnética/métodos , Espectrometria de Massas/métodos , Estrutura Molecular , Pós , Controle de Qualidade , Proteínas Recombinantes , Temperatura , Fatores de TempoRESUMO
Novel 2'-modified guanosine nucleosides were synthesized from inexpensive starting materials in 7-10 steps via hydroazidation or hydrocyanation reactions of the corresponding 2'-olefin. The antiviral effectiveness of the guanosine nucleosides was evaluated by converting them to the corresponding 5'-O-triphosphates (compounds 38-44) and testing their biochemical inhibitory activity against the wild-type NS5B polymerase.
Assuntos
Antivirais/síntese química , Nucleotídeos de Guanina/síntese química , Inibidores da Síntese de Ácido Nucleico/síntese química , Proteínas não Estruturais Virais/antagonistas & inibidores , Alcenos/síntese química , Azidas/síntese química , Hepacivirus/enzimologia , Proteínas não Estruturais Virais/químicaRESUMO
Starting from indole-based C-3 pyridone HCV NS5B polymerase inhibitor 2, structure-activity relationship (SAR) investigations of the indole N-1 benzyl moiety were performed. This study led to the discovery of irreversible inhibitors with p-fluoro-sulfone- or p-fluoro-nitro-substituted N-1 benzyl groups which achieved breakthrough replicon assay potency (EC(50) = 1 nM). The formation of a covalent bond with adjacent cysteine-366 thiol was was proved by mass spectroscopy and X-ray crystal structure studies. The C-5 ethyl C-2 carboxylic acid derivative 47 had an excellent oral area-under-the-curve (AUC) of 18 µM·h (10 mg/kg). Its oral exposure in monkeys and dogs was also very good. The NMR ALARM assay, mass spectroscopy experiments, in vitro counter screening, and toxicology assays demonstrated that the covalent bond formation between compound 47 and the protein was highly selective and specific. The overall excellent profile of 47 made it an interesting candidate for further investigation.
Assuntos
Antivirais/síntese química , Hepacivirus/efeitos dos fármacos , Indóis/síntese química , Nitrocompostos/síntese química , Sulfonas/síntese química , Proteínas não Estruturais Virais/antagonistas & inibidores , Animais , Antivirais/farmacocinética , Antivirais/farmacologia , Cristalografia por Raios X , Cães , Haplorrinos , Hepacivirus/enzimologia , Indóis/farmacocinética , Indóis/farmacologia , Modelos Moleculares , Estrutura Molecular , Testes de Mutagenicidade , Nitrocompostos/farmacocinética , Nitrocompostos/farmacologia , Piridonas/síntese química , Piridonas/farmacocinética , Piridonas/farmacologia , Ratos , Relação Estrutura-Atividade , Sulfonas/farmacocinética , Sulfonas/farmacologiaRESUMO
The 70% aqueous methanolic extract of the Peruvian plant Oryctanthus sp. was found to contain a novel saccharide of a diene alpha,omega-diacid. Compound 1 was identified as an inhibitor of the VEGF receptor. The structure of this compound was established based on NMR studies. Compound 1 inhibited ligand binding to the VEGF receptor with an IC50 of 5.0 microM.
Assuntos
Antineoplásicos/isolamento & purificação , Antineoplásicos/farmacologia , Magnolia/química , Receptores de Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Antineoplásicos/química , Concentração Inibidora 50 , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Receptores de Fatores de Crescimento do Endotélio Vascular/metabolismoRESUMO
The aqueous methanolic extract of a sea cucumber was found to contain two triterpene glycosides 1 and 2. The structures of 1 and 2 were established based on high-resolution NMR studies. Compounds 1 and 2 exhibited inhibitory activity (K(i)) of 30 and 5microM, respectively, in a chemokine receptor subtype 5 (CCR5) assay. Both compounds did not show any significant inhibition in a CXCR2 assay at 50microM, suggesting their selectivity for the CCR5 receptor.
Assuntos
Antagonistas dos Receptores CCR5 , Glicosídeos/farmacologia , Pepinos-do-Mar/química , Triterpenos/farmacologia , Animais , Glicosídeos/isolamento & purificação , Indicadores e Reagentes , Espectroscopia de Ressonância Magnética , Conformação Molecular , Espectrometria de Massas de Bombardeamento Rápido de Átomos , Extratos de Tecidos , Triterpenos/isolamento & purificaçãoRESUMO
The 70% aqueous methanol extract of the Peruvian plant Lippia alva (Verbenaceae) was found to contain three novel compounds, 1, 2, and 3, which were identified as inhibitors of the chemokine receptor CCR5. The structures of 1-3 were established based on extensive NMR studies. Compounds 1-3 inhibited CCR5 receptor signaling as measured by a calcium mobilization assay with IC(50) values of 5.5, 6.0, and 7.2 microg/mL, respectively.
Assuntos
Fármacos Anti-HIV/isolamento & purificação , Antagonistas dos Receptores CCR5 , Ciclopropanos/isolamento & purificação , Lactonas/isolamento & purificação , Lippia/química , Fármacos Anti-HIV/química , Fármacos Anti-HIV/farmacologia , Sinalização do Cálcio , Linhagem Celular Tumoral , Ciclopropanos/química , Ciclopropanos/farmacologia , Humanos , Lactonas/química , Lactonas/farmacologia , Espectroscopia de Ressonância Magnética , Extratos Vegetais/química , Relação Estrutura-AtividadeRESUMO
The 70% aqueous methanolic extract of the Peruvian plant Polygonum cuspidatum sp. was found to contain two novel phenolic saccharides 1 and 2, which were identified as inhibitors of the bacterial DNA primase enzyme. Structures of these two compounds were established based on high resolution NMR studies. Compound 1 and 2 inhibited the primase enzyme with an IC(50) of 4 and 5 microM, respectively.
Assuntos
Bactérias/enzimologia , DNA Primase/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Fallopia japonica/química , Inibidores Enzimáticos/química , Inibidores Enzimáticos/isolamento & purificação , Análise EspectralRESUMO
The 70% aq methanolic extract of the Peruvian plant Stylogne cauliflora was found to contain two novel oligophenolic compounds SCH 644343 (1) and SCH 644342 (2), which were identified as inhibitors of HCV NS3 protease. The structure of 1 and 2 was established based on high-resolution NMR studies. Compound 1 inhibited HCV NS3 protease with an IC(50) of 0.3 microM, while compound 2 showed an IC(50) of 0.8 microM.