RESUMO
HIV-1 replication has been inhibited by using a compound able to target the human cellular cofactor DEAD-box ATPase DDX3, essential for HIV-1 RNA nuclear export. This compound, identified by means of a computational protocol based on pharmacophoric modeling and molecular docking calculations, represents the first small molecule with such a mechanism of action and could lay the foundations for a pioneering approach for the treatment of HIV-1 infections.
Assuntos
RNA Helicases DEAD-box/antagonistas & inibidores , RNA Helicases DEAD-box/metabolismo , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , HIV-1/efeitos dos fármacos , HIV-1/enzimologia , Replicação Viral , Cristalografia por Raios X , RNA Helicases DEAD-box/química , Avaliação Pré-Clínica de Medicamentos , Humanos , Modelos Moleculares , Estrutura Terciária de ProteínaRESUMO
An efficient synthesis of the acid part of salvianolic acid E 2 is described. Compound 2 was obtained from vanillin in 10 steps and 21% overall yield. During the synthesis of 2 an unexpected 5-oxo-4b,9b-dihydroindano[1,2-b]benzofuran rac-12 was isolated. Both compounds together with the acid part of salvianolic acid D were active as HIV-1 integrase inhibitors at the submicromolar level. But they did not inhibit the replication of the virus on MT-4 cells.
Assuntos
Antivirais/síntese química , Antivirais/farmacologia , Química Farmacêutica/métodos , Flavonoides/síntese química , Flavonoides/farmacologia , Fenóis/síntese química , Fenóis/farmacologia , Extratos Vegetais/síntese química , Extratos Vegetais/farmacologia , Salvia/metabolismo , Benzaldeídos/química , Brometos/química , Linhagem Celular , Desenho de Fármacos , Inibidores Enzimáticos/farmacologia , Inibidores de Integrase de HIV/síntese química , Inibidores de Integrase de HIV/farmacologia , Humanos , Concentração Inibidora 50 , Inibidores de Integrase/farmacologia , Modelos Químicos , PolifenóisRESUMO
We have identified 1H-benzylindole analogues as a novel series of human immunodeficiency virus (HIV) integrase inhibitors with antiretroviral activities against different strains of HIV type 1 (HIV-1), HIV-2, and simian immunodeficiency virus strain MAC(251) [SIV(MAC(251))]. Molecular modeling and structure-activity relationship-based optimization resulted in the identification of CHI/1043 as the most potent congener. CHI/1043 inhibited the replication of HIV-1(III(B)) in MT-4 cells at a 50% effective concentration (EC(50)) of 0.60 microM, 70-fold below its cytotoxic concentration. Equal activities against HIV-1(NL4.3), HIV-2(ROD), HIV-2(EHO), and SIV(MAC(251)) were observed. CHI/1043 was equally active against virus strains resistant against inhibitors of reverse transcriptase or protease. Replication of both X4 and R5 strains in peripheral blood mononuclear cells was sensitive to the inhibitory effect of CHI/1043 (EC(50), 0.30 to 0.38 microM). CHI/1043 inhibited integrase strand transfer activity in oligonucleotide-based enzymatic assays at low micromolar concentrations. Time-of-addition experiments confirmed CHI/1043 to interfere with the viral replication cycle at the time of retroviral integration. Quantitative Alu PCR corroborated that the anti-HIV activity is based upon the inhibition of proviral DNA integration. An HIV-1 strain selected for 70 passages in the presence of CHI/1043 was evaluated genotypically and phenotypically. The mutations T66I and Q146K were present in integrase. Cross-resistance to other integrase strand transfer inhibitors, such as L-708,906, the naphthyridine analogue L-870,810, and the clinical drugs GS/9137 and MK-0518, was observed. In adsorption, distribution, metabolism, excretion, and toxicity studies, antiviral activity was strongly reduced by protein binding, and metabolization in human liver microsomes was observed. Transport studies with Caco cells suggest a low oral bioavailability.
Assuntos
Inibidores de Integrase de HIV/farmacologia , HIV/efeitos dos fármacos , Indóis/farmacologia , Integrases/metabolismo , Fármacos Anti-HIV/síntese química , Fármacos Anti-HIV/química , Fármacos Anti-HIV/farmacologia , Células CACO-2 , Linhagem Celular Tumoral , Avaliação Pré-Clínica de Medicamentos , Ensaio de Imunoadsorção Enzimática , HIV/enzimologia , HIV/genética , Inibidores de Integrase de HIV/síntese química , Inibidores de Integrase de HIV/química , Humanos , Indóis/síntese química , Indóis/química , Integrases/genética , Estrutura Molecular , Reação em Cadeia da Polimerase , Proteínas Virais/metabolismo , Replicação Viral/efeitos dos fármacosRESUMO
The identification of a novel hit compound as integrase binding inhibitor has been accomplished by means of virtual screening techniques. A small family of structurally related molecules has been synthesized and biologically evaluated with one of the compounds showing an IC(50)=12 microM.
Assuntos
Inibidores de Integrase de HIV/síntese química , Inibidores de Integrase de HIV/farmacologia , HIV-1/efeitos dos fármacos , Simulação por Computador , Bases de Dados Factuais , Avaliação Pré-Clínica de Medicamentos , HIV-1/enzimologia , Indicadores e Reagentes , Ligantes , Modelos MolecularesRESUMO
We describe the use of pharmacophore modeling as an efficient tool in the discovery of novel HIV-1 integrase (IN) inhibitors. A three-dimensional hypothetical model for the binding of diketo acid analogues to the enzyme was built by means of the Catalyst program. Using these models as a query for virtual screening, we found several compounds that contain the specified 3D patterns of chemical functions. Biological testing shows that our strategy was successful in searching for new structural leads as HIV-1 IN inhibitors.
Assuntos
Avaliação Pré-Clínica de Medicamentos/estatística & dados numéricos , Inibidores de Integrase de HIV/química , Inibidores de Integrase de HIV/farmacologia , Simulação por Computador , DNA Viral/efeitos dos fármacos , DNA Viral/genética , Bases de Dados Factuais , Desenho de Fármacos , HIV-1/efeitos dos fármacos , HIV-1/enzimologia , HIV-1/genética , Modelos Moleculares , Estrutura Molecular , Relação Estrutura-Atividade , Interface Usuário-ComputadorRESUMO
Limited information is available on the activity of antiretroviral drugs against human immunodeficiency virus type 2 (HIV-2) and simian immunodeficiency virus (SIV) strains to guide their use in treatment or prophylaxis. We evaluated the antiviral activity of 16 approved drugs and one experimental drug, AMD3100, against two wild-type HIV-2 (ROD and EHO) isolates, two strains of SIV (mac251 and B670), and two strains of simian-human immunodeficiency virus (SHIV) that contain the reverse transcriptase (RTSHIV) or envelope glycoprotein (SHIV89.6) of human immunodeficiency virus type 1 (HIV-1) in a SIV(mac239) background. Drug susceptibility was measured conventionally by the MT-4/MTT assay, and results were analysed as fold changes in 50% effective concentration (EC50) relative to the EC50 for HIV-1 (IIIB). The nucleoside reverse transcriptase inhibitors (NRTIs) zidovudine, lamivudine, stavudine, didanosine, zalcitabine and abacavir as well as the nucleotide reverse transcriptase inhibitor tenofovir retained full activity against all six viruses except for SIV and SHIV89.6 that showed low-level resistance to didanosine. The protease inhibitors (PIs) ritonavir, indinavir, saquinavir and nelfinavir were found to be active against some HIV-2 or SIV strains. However, a significant reduction in susceptibility was seen with indinavir against SHIV89.6 (3.3-fold), and with amprenavir against HIV-2(ROD) (8.8-fold). All viruses except for RTSHIV showed a >200-fold decrease in susceptibility for the non-nucleoside reverse transcriptase inhibitors (NNRTIs) nevirapine, delavirdine and efavirenz, indicating high-level resistance. AMD3100, a CXCR4 antagonist, was active against HIV-2 and SHIV89.6, a finding consistent with the use of the CXCR4 co-receptor by these isolates, but was inactive against SIV strains. In contrast, enfuvirtide (T-20) was active against SHIV89.6 but had reduced inhibitory activity against both HIV-2 and SIV strains predicting little therapeutic value against these viruses. These findings support the use of NRTIs, tenofovir, but not NNRTIs, for treating HIV-2-infected persons or for prophylaxis against HIV-2 and SIV. The clinical significance of the low-level resistance of HIV-2 and SIV to some PIs is unclear. Co-receptor antagonists such as AMD3100 show promising anti-HIV-2 therapeutic modalities. Both AMD3100 and enfuvirtide could be used for prophylaxis against SHIV89.6.
Assuntos
Fármacos Anti-HIV/farmacologia , HIV-2/efeitos dos fármacos , HIV/efeitos dos fármacos , Vírus da Imunodeficiência Símia/efeitos dos fármacos , Sequência de Aminoácidos , Animais , Sequência Consenso , Transcriptase Reversa do HIV/genética , Humanos , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Inibidores de Proteases/farmacologia , DNA Polimerase Dirigida por RNA/genética , Inibidores da Transcriptase Reversa/farmacologia , Alinhamento de Sequência , Homologia de Sequência de AminoácidosRESUMO
Leaf and (unripe and ripe) berry essential oils of Juniperus oxycedrus ssp. badia (H. Gay) Debeaux grown wild in Spain have been analysed by capillary GC and GC-MS in combination with retention indices. A seasonal investigation of both leaf and berry oils was also performed. Among the approximately 80 constituents investigated (representing 90-98% of the oils) 60-68 were identified (80-97% of the oil composition). The leaf oils were mainly composed of alpha-pinene (40-57%) and manoyl oxide (5-10%). The (unripe) berry oils were dominated by alpha-pinene (65%) with moderate amounts of myrcene, limonene, germacrene D or gamma-muurolene. Several differences in yields and chemical composition from a qualitative and quantitative point of view were detected when comparing all the oil samples analysed. In addition, two oil samples were examined, but found inactive, against the replication of HIV-1(III(B)) and HIV-2(ROD) in cell culture, whereas the samples were toxic for the cells at a 50% cytotoxic concentration of 106 and 123 microg/ml, respectively.