RESUMEN
A novel HIV protease inhibitor was designed using a morpholine core as the aspartate binding group. Analysis of the crystal structure of the initial lead bound to HIV protease enabled optimization of enzyme potency and antiviral activity. This afforded a series of potent orally bioavailable inhibitors of which MK-8718 was identified as a compound with a favorable overall profile.
RESUMEN
MK-6186 is a novel nonnucleoside reverse transcriptase inhibitor (NNRTI) which displays subnanomolar potency against wild-type (WT) virus and the two most prevalent NNRTI-resistant RT mutants (K103N and Y181C) in biochemical assays. In addition, it showed excellent antiviral potency against K103N and Y181C mutant viruses, with fold changes (FCs) of less than 2 and 5, respectively. When a panel of 12 common NNRTI-associated mutant viruses was tested with MK-6186, only 2 relatively rare mutants (Y188L and V106I/Y188L) were highly resistant, with FCs of >100, and the remaining viruses showed FCs of <10. Furthermore, a panel of 96 clinical virus isolates with NNRTI resistance mutations was evaluated for susceptibility to NNRTIs. The majority (70%) of viruses tested displayed resistance to efavirenz (EFV), with FCs of >10, whereas only 29% of the mutant viruses displayed greater than 10-fold resistance to MK-6186. To determine whether MK-6186 selects for novel resistance mutations, in vitro resistance selections were conducted with one isolate each from subtypes A, B, and C under low-multiplicity-of-infection (MOI) conditions. The results showed a unique mutation development pattern in which L234I was the first mutation to emerge in the majority of the experiments. In resistance selection under high-MOI conditions with subtype B virus, V106A was the dominant mutation detected in the breakthrough viruses. More importantly, mutant viruses selected by MK-6186 showed FCs of <10 against EFV or etravirine (ETR), and the mutant viruses containing mutations selected by EFV or ETR were sensitive to MK-6186 (FCs of <10).
Asunto(s)
Fármacos Anti-VIH/farmacología , VIH-1/enzimología , Inhibidores de la Transcriptasa Inversa/farmacología , Alquinos , Benzoxazinas/farmacología , Ciclopropanos , Transcriptasa Inversa del VIH/antagonistas & inhibidores , Transcriptasa Inversa del VIH/genética , VIH-1/genética , MutaciónRESUMEN
Next generation NNRTIs are sought which possess both broad spectrum antiviral activity against key mutant strains and a high genetic barrier to the selection of new mutant viral strains. Pyridones were evaluated as an acyclic conformational constraint to replace the aryl ether core of MK-4965 (1) and the more rigid indazole constraint of MK-6186 (2). The resulting pyridone compounds are potent inhibitors of HIV RT and have antiviral activity in cell culture that is superior to other next generation NNRTI's.
Asunto(s)
Transcriptasa Inversa del VIH/antagonistas & inhibidores , Piridonas/química , Inhibidores de la Transcriptasa Inversa/síntesis química , Sitios de Unión , Línea Celular , Simulación por Computador , Diseño de Fármacos , Activación Enzimática/efectos de los fármacos , VIH/enzimología , Transcriptasa Inversa del VIH/metabolismo , Humanos , Estructura Terciaria de Proteína , Pirazoles/química , Piridinas/química , Piridonas/síntesis química , Piridonas/farmacología , Inhibidores de la Transcriptasa Inversa/química , Inhibidores de la Transcriptasa Inversa/farmacologíaRESUMEN
Highly active antiretroviral therapy (HAART) significantly reduces human immunodeficiency virus (HIV) viral load and has led to a dramatic decrease in acquired immunodeficiency syndrome (AIDS) related mortality. Despite this success, there remains a critical need for new HIV therapies to address the emergence of drug resistant viral strains. Next generation NNRTIs are sought that are effective against these mutant forms of the HIV virus. The bound conformations of our lead inhibitors, MK-1107 (1) and MK-4965 (2), were divergent about the oxymethylene linker, and each of these conformations was rigidified using two isomeric cyclic constraints. The constraint derived from the bioactive conformation of 2provided novel, highly potent NNRTIs that possess broad spectrum antiviral activity and good pharmacokinetic profiles. Systematic SAR led to the identification of indazole as the optimal conformational constraint to provide MK-6186 (3) and MK-7445 (6). Despite their reduced flexibility, these compounds had potency comparable to that of the corresponding acyclic ethers in both recombinant enzyme and cell based assays against both the wild-type and the clinically relevant mutant strains.
Asunto(s)
Fármacos Anti-VIH/síntesis química , Imidazoles/síntesis química , Indazoles/síntesis química , Pirazoles/síntesis química , Inhibidores de la Transcriptasa Inversa/síntesis química , Triazoles/síntesis química , Animales , Fármacos Anti-VIH/farmacocinética , Fármacos Anti-VIH/farmacología , Células Cultivadas , Perros , Transcriptasa Inversa del VIH/química , Transcriptasa Inversa del VIH/metabolismo , VIH-1/efectos de los fármacos , VIH-1/genética , VIH-1/aislamiento & purificación , Humanos , Imidazoles/farmacocinética , Imidazoles/farmacología , Indazoles/farmacocinética , Indazoles/farmacología , Modelos Moleculares , Conformación Molecular , Mutación , Nitrilos/síntesis química , Nitrilos/farmacocinética , Nitrilos/farmacología , Nitrobencenos/síntesis química , Nitrobencenos/farmacocinética , Nitrobencenos/farmacología , Pirazoles/farmacocinética , Pirazoles/farmacología , Ratas , Ratas Sprague-Dawley , Proteínas Recombinantes/química , Inhibidores de la Transcriptasa Inversa/farmacocinética , Inhibidores de la Transcriptasa Inversa/farmacología , Relación Estructura-Actividad , Termodinámica , Triazoles/farmacocinética , Triazoles/farmacologíaRESUMEN
AIDS is produced by HIV-induced infections. HIV integrase is an important enzyme as it is critical for the integration of the HIV genome into that of the host cell. This complex process is exclusively carried out by a viral enzyme not found in the host cell. Therefore, this protein represents a safe target for the development of single or combined anti-HIV therapy. Integramide A is a 16-mer long, effective peptaib inhibitor of HIV-1 integrase. We have previously described a versatile synthetic strategy in solution to afford this natural compound and its diastereomer at positions 14 and 15. We also found that both peptides display a significant inhibitory activity. Here, we present our data on the synthesis in solution, in-depth conformational analysis, and biological activity against HIV-integrase of the analogs of the two above mentioned peptides in which all of the three (2S,4R)-Hyp residues at positions 2, 9, and 13 are replaced by L-Pro. This study definitely confirms that the mixed α-/3(10) - helical conformation of natural integramide A plays a key role in its mechanism of inhibition. Moreover, our data provide evidence that the amphipathic character of this helical structure is not required for the activity of integramide A against HIV-1 integrase. These observations will hopefully help us to further clarify the precise mechanism of inhibition of this interesting peptaib and to identify shorter peptide sequences active against HIV-1 integrase.
Asunto(s)
Inhibidores de Integrasa VIH/química , Inhibidores de Integrasa VIH/farmacología , Oligopéptidos/química , Oligopéptidos/farmacología , Secuencia de Aminoácidos , Dicroismo Circular , VIH-1/efectos de los fármacos , VIH-1/enzimología , Humanos , Hidroxiprolina/química , Espectroscopía de Resonancia Magnética , Datos de Secuencia Molecular , Prolina/química , Conformación Proteica , Estructura Secundaria de Proteína , Espectroscopía Infrarroja por Transformada de Fourier , Relación Estructura-ActividadRESUMEN
Studies were conducted to investigate mutation pathways among subtypes A, B, and C of human immunodeficiency virus type 1 (HIV-1) during resistance selection with nonnucleoside reverse transcriptase inhibitors (NNRTIs) in cell culture under low-multiplicity of infection (MOI) conditions. The results showed that distinct pathways were selected by different virus subtypes under increasing selective pressure of NNRTIs. F227C and Y181C were the major mutations selected by MK-4965 in subtype A and C viruses during resistance selection. With efavirenz (EFV), F227C and V106M were the major mutations responsible for viral breakthrough in subtype A viruses, whereas a single pathway (G190A/V106M) accounted for mutation development in subtype C viruses. Y181C was the dominant mutation in the resistance selection with etravirine (ETV) in subtype A, and E138K/H221Y were the mutations detected in the breakthrough viruses from subtype C viruses with ETV. In subtype B viruses, on the other hand, known NNRTI-associated mutations (e.g., Y181C, P236L, L100I, V179D, and K103N) were selected by the NNRTIs. The susceptibility of the subtype A and B mutant viruses to NNRTIs was determined in order to gain insight into the potential mechanisms of mutation development. Collectively, these results suggest that minor differences may exist in conformation of the residues within the NNRTI binding pocket (NNRTIBP) of reverse transcriptase (RT) among the three subtypes of viruses. Thus, the interactions between NNRTIs and the residues in the NNRTIBPs of different subtypes may not be identical, leading to distinct mutation pathways during resistance selection in cell culture.
Asunto(s)
Farmacorresistencia Viral/genética , Transcriptasa Inversa del VIH/antagonistas & inhibidores , VIH-1/efectos de los fármacos , Inhibidores de la Transcriptasa Inversa/farmacología , Alquinos , Benzoxazinas/química , Benzoxazinas/farmacología , Línea Celular , Ciclopropanos , VIH-1/genética , Humanos , Estructura Molecular , Mutación , Nitrilos , Pirazoles/química , Pirazoles/farmacología , Piridazinas/química , Piridazinas/farmacología , Piridinas/química , Piridinas/farmacología , Pirimidinas , Replicación Viral/efectos de los fármacosRESUMEN
Biaryl ethers were recently reported as potent NNRTIs. Herein, we disclose a detailed effort to modify the previously reported compound 1. We have designed and synthesized a series of novel pyrazole derivatives as a surrogate for pyrazolopyridine motif that were potent inhibitors of HIV-1 RT with nanomolar intrinsic activity on the WT and key mutant enzymes and potent antiviral activity in infected cells.
Asunto(s)
Fármacos Anti-VIH/química , Éteres/química , Transcriptasa Inversa del VIH/antagonistas & inhibidores , Pirazoles/química , Piridinas/química , Inhibidores de la Transcriptasa Inversa/química , Regulación Alostérica , Animales , Fármacos Anti-VIH/síntesis química , Fármacos Anti-VIH/farmacocinética , Perros , Éteres/síntesis química , Éteres/farmacocinética , Transcriptasa Inversa del VIH/genética , Transcriptasa Inversa del VIH/metabolismo , Humanos , Mutación , Pirazoles/síntesis química , Pirazoles/farmacocinética , Piridinas/síntesis química , Piridinas/farmacocinética , Ratas , Inhibidores de la Transcriptasa Inversa/síntesis química , Inhibidores de la Transcriptasa Inversa/farmacocinética , Relación Estructura-ActividadRESUMEN
Traditional phenotypic assays used to assess the susceptibility of mutant human immunodeficiency virus type-1 (HIV-1) obtained from infected patients or from resistance selection to antiviral agents in cell culture are rather tedious and time consuming. To improve the efficiency of this process, a novel method was developed in which mutant viruses are captured with magnetic nano-beads and used to infect gag-GFP reporter cells to evaluate the extent of resistance conferred by the mutant viruses against antiviral agents. The optimal timing for measuring the inhibitory potencies of antiviral agents was found to be day 3 post-infection for integrase strand transfer inhibitors and protease inhibitors and day 4 for non-nucleoside reverse transcriptase inhibitors. Comparable EC(50) values were obtained when bead-captured breakthrough virus from in vitro resistance selection experiments and its matched site-directed mutagenesis virus were tested side by side in this assay. This assay protocol was also employed to evaluate the inhibitor susceptibility of breakthrough viruses collected from resistance selections that were conducted in the presence of increasing concentrations of an HIV-1 protease inhibitor. Taken together, these findings suggest that a rapid, sensitive, non-invasive, and homogeneous phenotypic assay has been developed for assessing the antiviral agent susceptibility of mutant viruses that emerge from in vitro resistance selection studies.
Asunto(s)
Fármacos Anti-VIH/farmacología , VIH-1/efectos de los fármacos , Pruebas de Sensibilidad Microbiana/métodos , Bioensayo , Línea Celular , Farmacorresistencia Viral Múltiple/genética , Farmacorresistencia Viral/genética , Proteínas Fluorescentes Verdes/genética , VIH-1/genética , Humanos , Microesferas , Mutagénesis Sitio-Dirigida , Mutación , Transfección , Productos del Gen gag del Virus de la Inmunodeficiencia Humana/genéticaRESUMEN
Integramide A is a 16-amino acid peptide inhibitor of the enzyme HIV-1 integrase. We have recently reported that the absolute stereochemistries of the dipeptide sequence near the C terminus are L-Iva(14)-D-Iva(15). Herein, we describe the syntheses of the natural compound and its D-Iva(14)-L-Iva(15) diastereomer, and the results of their chromatographic/mass spectrometric analyses. We present the conformational analysis of the two compounds and some of their synthetic intermediates of different main-chain length in the crystal state (by X-ray diffraction) and in solvents of different polarities (using circular dichroism, FTIR absorption, and 2D NMR techniques). These data shed light on the mechanism of inhibition of HIV-1 integrase, which is an important target for anti-HIV therapy.
Asunto(s)
Dipéptidos/química , Inhibidores de Integrasa VIH/síntesis química , Inhibidores de Integrasa VIH/farmacología , Integrasa de VIH/metabolismo , VIH-1/efectos de los fármacos , Oligopéptidos/síntesis química , Oligopéptidos/farmacología , Péptidos/síntesis química , Secuencia de Aminoácidos , Dicroismo Circular , Inhibidores de Integrasa VIH/química , Humanos , Espectroscopía de Resonancia Magnética , Modelos Moleculares , Conformación Molecular , Datos de Secuencia Molecular , Oligopéptidos/química , Fragmentos de Péptidos , Péptidos/química , Estereoisomerismo , Difracción de Rayos XRESUMEN
Biaryl ethers were recently reported as potent NNRTIs. Herein we disclose a detailed SAR study that led to the biaryl ether 6. This compound possessed excellent potency against WT RT and key clinically observed RT mutants and had an excellent pharmacokinetic profile in rats, dogs, and rhesus macaques. The compound also exhibited a clean safety profile in preclinical safety studies.
Asunto(s)
Éteres/química , Éteres/farmacología , Transcriptasa Inversa del VIH/antagonistas & inhibidores , Transcriptasa Inversa del VIH/genética , VIH-1/efectos de los fármacos , VIH-1/genética , Mutación , Animales , Línea Celular , Perros , Éteres/síntesis química , Éteres/farmacocinética , VIH-1/enzimología , Humanos , Macaca mulatta , Nucleósidos/química , Ratas , Inhibidores de la Transcriptasa Inversa/síntesis química , Inhibidores de la Transcriptasa Inversa/química , Inhibidores de la Transcriptasa Inversa/farmacocinética , Inhibidores de la Transcriptasa Inversa/farmacología , Relación Estructura-ActividadRESUMEN
Non-nucleoside reverse transcriptase inhibitors (NNRTIs) are key elements of multidrug regimens, called HAART (Highly Active Antiretroviral Therapy), that are used to treat HIV-1 infections. Elucidation of the structure-activity relationships of the thiocarbamate moiety of the previous published lead compound 2 provided a series of novel tetrahydroquinoline derivatives as potent inhibitors of HIV-1 RT with nanomolar intrinsic activity on the WT and key mutant enzymes and potent antiviral activity in infected cells. The SAR optimization, mutation profiles, preparation of compounds, and pharmacokinetic profile of compounds are described.
Asunto(s)
Fármacos Anti-VIH/química , Transcriptasa Inversa del VIH/antagonistas & inhibidores , Quinolinas/química , Inhibidores de la Transcriptasa Inversa/química , Sitio Alostérico , Fármacos Anti-VIH/síntesis química , Fármacos Anti-VIH/farmacología , Sitios de Unión , Cristalografía por Rayos X , Transcriptasa Inversa del VIH/metabolismo , Conformación Molecular , Proteínas Mutantes/antagonistas & inhibidores , Proteínas Mutantes/metabolismo , Quinolinas/síntesis química , Quinolinas/farmacología , Inhibidores de la Transcriptasa Inversa/síntesis química , Inhibidores de la Transcriptasa Inversa/farmacología , Relación Estructura-Actividad , Tiocarbamatos/química , Tiocarbamatos/farmacologíaAsunto(s)
Productos Biológicos/química , Productos Biológicos/farmacología , Inhibidores de Integrasa VIH/química , Inhibidores de Integrasa VIH/farmacología , Integrasa de VIH/metabolismo , Oligopéptidos/química , Oligopéptidos/farmacología , Secuencia de Aminoácidos , Productos Biológicos/síntesis química , Cromatografía Líquida de Alta Presión , Inhibidores de Integrasa VIH/síntesis química , Espectroscopía de Resonancia Magnética , Datos de Secuencia Molecular , Oligopéptidos/síntesis química , EstereoisomerismoRESUMEN
Non-nucleoside reverse transcriptase inhibitors (NNRTIs) have been shown to be a key component of highly active antiretroviral therapy (HAART). The use of NNRTIs has become part of standard combination antiviral therapies producing clinical outcomes with efficacy comparable to other antiviral regimens. There is, however, a critical issue with the emergence of clinical resistance, and a need has arisen for novel NNRTIs with a broad spectrum of activity against key HIV-1 RT mutations. Using a combination of traditional medicinal chemistry/SAR analyses, crystallography, and molecular modeling, we have designed and synthesized a series of novel, highly potent NNRTIs that possess broad spectrum antiviral activity and good pharmacokinetic profiles. Further refinement of key compounds in this series to optimize physical properties and pharmacokinetics has resulted in the identification of 8e (MK-4965), which has high levels of potency against wild-type and key mutant viruses, excellent oral bioavailability and overall pharmacokinetics, and a clean ancillary profile.
Asunto(s)
Transcriptasa Inversa del VIH/antagonistas & inhibidores , VIH-1/efectos de los fármacos , VIH-1/enzimología , Pirazoles/síntesis química , Pirazoles/farmacología , Piridinas/síntesis química , Piridinas/farmacología , Inhibidores de la Transcriptasa Inversa/síntesis química , Inhibidores de la Transcriptasa Inversa/farmacología , Administración Oral , Animales , Compuestos de Bromina/síntesis química , Compuestos de Bromina/química , Cristalografía por Rayos X , Evaluación Preclínica de Medicamentos , Transcriptasa Inversa del VIH/química , Transcriptasa Inversa del VIH/genética , Transcriptasa Inversa del VIH/metabolismo , VIH-1/genética , Modelos Moleculares , Estructura Molecular , Mutación/genética , Nucleósidos/química , Nucleósidos/farmacología , Pirazoles/química , Piridinas/química , Ratas , Ratas Sprague-Dawley , Inhibidores de la Transcriptasa Inversa/química , Relación Estructura-ActividadRESUMEN
HIV-1 integrase catalyzes the insertion of viral DNA into the genome of the host cell. Integrase inhibitor N-(4-fluorobenzyl)-8-hydroxy-1,6-naphthyridine-7-carboxamide selectively inhibits the strand transfer process of integration. 4-Substituted pyrrolidinones possessing various groups on the pyrrolidinone nitrogen were introduced at the 5-position of the naphthyridine scaffold. These analogs exhibit excellent activity against viral replication in a cell-based assay. The preparation of these compounds was enabled by a three-step, two-pot reaction sequence from a common butenolide intermediate.
Asunto(s)
Fármacos Anti-VIH/síntesis química , Fármacos Anti-VIH/farmacología , Inhibidores de Integrasa VIH/síntesis química , Inhibidores de Integrasa VIH/farmacología , Integrasa de VIH/metabolismo , VIH-1/efectos de los fármacos , Naftiridinas/síntesis química , Naftiridinas/farmacología , Administración Oral , Animales , Fármacos Anti-VIH/química , Inhibidores de Integrasa VIH/química , Estructura Molecular , Naftiridinas/química , Ratas , Relación Estructura-ActividadRESUMEN
A series of 10-hydroxy-7,8-dihydropyrazino[1',2':1,5]pyrrolo[2,3-d]pyridazine-1,9(2H,6H)-diones was synthesized and tested for their inhibition of HIV-1 replication in cell culture. Structure-activity studies indicated that high antiviral potency against wild-type virus as well as viruses containing integrase mutations that confer resistance to three different structural classes of integrase inhibitors could be achieved by incorporation of small aliphatic groups at certain positions on the core template. An optimal compound from this study, 16, inhibits integrase strand-transfer activity with an IC(50) value of 10 nM, inhibits HIV-1 replication in cell culture with an IC(95) value of 35 nM in the presence of 50% normal human serum, and displays modest pharmacokinetic properties in rats (i.v. t(1/2)=5.3 h, F=17%).
Asunto(s)
Química Farmacéutica/métodos , Integrasa de VIH/síntesis química , Integrasa de VIH/farmacología , Integrasas/genética , Mutación , Administración Oral , Animales , Antivirales/farmacología , Disponibilidad Biológica , Diseño de Fármacos , Humanos , Concentración 50 Inhibidora , Modelos Químicos , Ratas , Relación Estructura-Actividad , Replicación ViralRESUMEN
Using a combination of traditional Medicinal Chemistry/SAR analysis, crystallography, and molecular modeling, we have designed and synthesized a series of novel, highly potent NNRTIs that possess broad antiviral activity against a number of key clinical mutations.
Asunto(s)
Fármacos Anti-VIH/farmacología , Diseño de Fármacos , Éteres/química , VIH-1/efectos de los fármacos , Inhibidores de la Transcriptasa Inversa/farmacología , Replicación Viral/efectos de los fármacos , Fármacos Anti-VIH/síntesis química , Sitios de Unión , Cristalografía por Rayos X , Farmacorresistencia Viral , VIH-1/enzimología , VIH-1/genética , Modelos Químicos , Mutación , Nucleósidos/química , Inhibidores de la Transcriptasa Inversa/síntesis química , Relación Estructura-Actividad , Replicación Viral/fisiologíaRESUMEN
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.
Asunto(s)
Aminopiridinas/síntesis química , Azepinas/síntesis química , Compuestos Bicíclicos Heterocíclicos con Puentes/síntesis química , Inhibidores de Integrasa VIH/síntesis química , Integrasa de VIH/metabolismo , Pirimidinonas/síntesis química , Administración Oral , Aminopiridinas/farmacocinética , Aminopiridinas/farmacología , Animales , Azepinas/farmacocinética , Azepinas/farmacología , Disponibilidad Biológica , Compuestos Bicíclicos Heterocíclicos con Puentes/farmacocinética , Compuestos Bicíclicos Heterocíclicos con Puentes/farmacología , Línea Celular , Perros , Integrasa de VIH/genética , Inhibidores de Integrasa VIH/farmacocinética , Inhibidores de Integrasa VIH/farmacología , VIH-1/efectos de los fármacos , Humanos , Macaca mulatta , Microsomas Hepáticos/metabolismo , Pirimidinonas/farmacocinética , Pirimidinonas/farmacología , Ratas , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
Reverse transcriptase (RT) plays an essential role in the HIV-1 replication process, which converts a single-strand RNA into a double-strand DNA via polymerase and RNase H activities. Therefore, inhibition of RT has been one of the primary therapeutic strategies for suppressing the replication of HIV-1. To facilitate the process of discovering the next generation of antiretroviral agents, this study presents a highly sensitive and nonradioactive RT polymerase assay that is based on electrochemiluminescence (ECL) technology, where a ruthenylated dUTP (Ru-dUTP) is employed as one of the dNTPs. The concentration of the RT enzymes required for the assay can be as low as 1 pM, enabling us to evaluate inhibitors with low picomolar potency. More importantly, the assay is capable of detecting endogenous RT activity in cell-free viruses. Therefore, the assay was applied to monitor the development of resistance mutation(s) by viruses under the treatment with a non-nucleoside reverse transcriptase inhibitor (NNRTI) in cell culture. The magnitude of resistance of the resulting mutant viruses was assessed directly by the assay, eliminating the need for cloning, expressing, and purifying the RT mutants.
Asunto(s)
Farmacorresistencia Viral , VIH-1/efectos de los fármacos , Mediciones Luminiscentes/métodos , ADN Polimerasa Dirigida por ARN/metabolismo , Inhibidores de la Transcriptasa Inversa/farmacología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Alquinos , Benzoxazinas/farmacología , Línea Celular , Ciclopropanos , Delavirdina/farmacología , Electroquímica/métodos , VIH-1/genética , Humanos , Nevirapina/farmacología , Compuestos Organometálicos/metabolismo , ADN Polimerasa Dirigida por ARN/genéticaRESUMEN
A series of 4-oxo-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine-2-carboxamides was synthesized and tested for their inhibition of HIV-1 integrase catalytic activity and HIV-1 replication in cells. Structure-activity studies around lead compound 5 indicated that a coplanar relationship of metal-binding heteroatoms provides optimal binding to the integrase active site. Identification of potency-enhancing substituents and adjustments in lipophilicity provided 17b which inhibits integrase-catalyzed strand transfer with an IC(50) value of 74 nM and inhibits HIV-1 replication in cell culture in the presence of 50% normal human serum with an IC(95) value of 63 nM.
Asunto(s)
Inhibidores de Integrasa VIH/química , Inhibidores de Integrasa VIH/farmacología , Integrasa de VIH/efectos de los fármacos , Pirazinas/química , Pirazinas/farmacología , Catálisis , Línea Celular , Integrasa de VIH/metabolismo , Inhibidores de Integrasa VIH/síntesis química , VIH-1/enzimología , VIH-1/fisiología , Pirazinas/síntesis química , Relación Estructura-Actividad , Replicación Viral/efectos de los fármacosRESUMEN
A series of potent novel 8-hydroxy-3,4-dihydropyrrolo[1,2-a]pyrazine-1(2H)-one HIV-1 integrase inhibitors was identified. These compounds inhibited the strand transfer process of HIV-1 integrase and viral replication in cells. Compound 12 is active against replication of HIV-1 in cell culture with a CIC(95) of 0.31microM. Further SAR exploration led to the preparation of pseudosymmetrical tricyclic pyrrolopyrazine inhibitors 23 and 24 with further improvement in antiviral activity.