RESUMO
The severe acute respiratory syndrome (SARS) coronavirus 3CL protease is an attractive target for the development of anti-SARS drugs. In this paper, cinanserin (1) analogs were synthesized and tested for the inhibitory activities against SARS-coronavirus (CoV) 3CL protease by fluorescence resonance energy transfer (FRET) assay. Four analogs show significant activities, especially compound 26 with an IC(50) of 1.06 microM.
Assuntos
Antivirais/farmacologia , Cinanserina/análogos & derivados , Cinanserina/farmacologia , Inibidores de Proteases/farmacologia , Antagonistas da Serotonina/farmacologia , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave/enzimologia , Proteínas Virais/antagonistas & inibidores , Antivirais/síntese química , Cinanserina/síntese química , Proteases 3C de Coronavírus , Cisteína Endopeptidases , Desenho de Fármacos , Transferência Ressonante de Energia de Fluorescência , Indicadores e Reagentes , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Inibidores de Proteases/síntese química , Antagonistas da Serotonina/síntese química , Espectrofotometria Infravermelho , Relação Estrutura-AtividadeRESUMO
Novel potential human immunodeficiency virus (HIV) protease inhibitors were designed by a combination of nelfinavir and amprenavir motifs. The designed compounds were prepared by a facile synthetic route and their stereochemistry was further confirmed by a stereospecific synthesis from commercially available (S)-2-oxiranylmethyl m-nitrobenzenesulfonate. All compounds were tested for their ability in inhibiting HIV type 1 protease activity with the published method of reference 19. Derivatives 1a--u exhibited moderate to significant inhibitory activities in preliminary bioassay. The best compound 1a has IC50 value of 0.02 microM, comparable to that of amprenavir. A docking study on compounds 1a--u was performed using the published X-ray crystal structure of HIV type 1 protease, all compounds bound to the HIV type 1 protease in an extended conformation and the scaffoldings of the binding conformations could be aligned quite well. Comparative molecular field analysis (CoMFA) study was performed to explore the specific contributions of electrostatic and steric effects in the binding of these new compounds to HIV type 1 protease and a predictive CoMFA model was built with thirteen compounds as training set. Test analysis of other five compounds as test set demonstrated that the CoMFA model has strong predictive ability to this series of compounds. It will be very useful to further optimize the designed inhibitors.
Assuntos
Fármacos Anti-HIV/síntese química , Inibidores da Protease de HIV/síntese química , HIV/efeitos dos fármacos , Propanolaminas/síntese química , Fármacos Anti-HIV/farmacologia , Fármacos Anti-HIV/uso terapêutico , Cristalografia por Raios X , Desenho de Fármacos , Infecções por HIV/tratamento farmacológico , Inibidores da Protease de HIV/farmacologia , Inibidores da Protease de HIV/uso terapêutico , Humanos , Concentração Inibidora 50 , Propanolaminas/farmacologia , Propanolaminas/uso terapêutico , Relação Estrutura-AtividadeAssuntos
Fármacos Anti-HIV/química , Inibidores da Protease de HIV/química , 4-Hidroxicumarinas/síntese química , 4-Hidroxicumarinas/química , 4-Hidroxicumarinas/farmacologia , Fármacos Anti-HIV/síntese química , Fármacos Anti-HIV/farmacologia , Sulfato de Atazanavir , Inibidores da Protease de HIV/síntese química , Inibidores da Protease de HIV/farmacologia , Humanos , Estrutura Molecular , Oligopeptídeos/síntese química , Oligopeptídeos/química , Oligopeptídeos/farmacologia , Piridinas/síntese química , Piridinas/química , Piridinas/farmacologia , Saquinavir/síntese química , Saquinavir/química , Saquinavir/farmacologia , Ureia/análogos & derivados , Ureia/síntese química , Ureia/química , Ureia/farmacologiaRESUMO
The 3C-like proteinase (3CLpro) of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is one of the most promising targets for anti-SARS-CoV drugs due to its crucial role in the viral life cycle. In this study, a database containing structural information of more than 8,000 existing drugs was virtually screened by a docking approach to identify potential binding molecules of SARS-CoV 3CLpro. As a target for screening, both a homology model and the crystallographic structure of the binding pocket of the enzyme were used. Cinanserin (SQ 10,643), a well-characterized serotonin antagonist that has undergone preliminary clinical testing in humans in the 1960s, showed a high score in the screening and was chosen for further experimental evaluation. Binding of both cinanserin and its hydrochloride to bacterially expressed 3CLpro of SARS-CoV and the related human coronavirus 229E (HCoV-229E) was demonstrated by surface plasmon resonance technology. The catalytic activity of both enzymes was inhibited with 50% inhibitory concentration (IC50) values of 5 microM, as tested with a fluorogenic substrate. The antiviral activity of cinanserin was further evaluated in tissue culture assays, namely, a replicon system based on HCoV-229E and quantitative test assays with infectious SARS-CoV and HCoV-229E. All assays revealed a strong inhibition of coronavirus replication at nontoxic drug concentrations. The level of virus RNA and infectious particles was reduced by up to 4 log units, with IC50 values ranging from 19 to 34 microM. These findings demonstrate that the old drug cinanserin is an inhibitor of SARS-CoV replication, acting most likely via inhibition of the 3CL proteinase.