RESUMO
Analysis of the HIV protease gene from the plasma of HIV-infected patients revealed substitutions at nine different codons selected in response to monotherapy with the protease inhibitor ritonavir. Mutants at valine-82, although insufficient to confer resistance, appeared first in most patients. Significant phenotypic resistance required multiple mutations in HIV protease, which emerged subsequently in an ordered, stepwise fashion. The appearance of resistance mutations was delayed in patients with higher plasma levels of ritonavir. Early mutants retained susceptibility to structurally diverse protease inhibitors, suggesting that dual protease inhibitor therapy might increase the duration of viral suppression.
Assuntos
Inibidores da Protease de HIV/farmacologia , Protease de HIV/genética , HIV/efeitos dos fármacos , Mutação , Tiazóis/farmacologia , Valina/análogos & derivados , Códon , Genótipo , HIV/enzimologia , HIV/genética , Infecções por HIV/sangue , Humanos , Fenótipo , Ritonavir , Valina/genética , Valina/farmacologiaRESUMO
A two-fold (C2) symmetric inhibitor of the protease of human immunodeficiency virus type-1 (HIV-1) has been designed on the basis of the three-dimensional symmetry of the enzyme active site. The symmetric molecule inhibited both protease activity and acute HIV-1 infection in vitro, was at least 10,000-fold more potent against HIV-1 protease than against related enzymes, and appeared to be stable to degradative enzymes. The 2.8 angstrom crystal structure of the inhibitor-enzyme complex demonstrated that the inhibitor binds to the enzyme in a highly symmetric fashion.
Assuntos
Endopeptidases/metabolismo , Produtos do Gene pol/metabolismo , HIV-1/enzimologia , Inibidores de Proteases/farmacologia , Álcoois Açúcares/farmacologia , Valina/análogos & derivados , Sequência de Aminoácidos , Sítios de Ligação , Desenho de Fármacos , Protease de HIV , Cinética , Modelos Moleculares , Dados de Sequência Molecular , Conformação Proteica , Valina/farmacologiaRESUMO
BACKGROUND AND PURPOSE: Current nonhuman primate stroke models are limited by either stroke variability or survivability. A new nonhuman primate stroke model was developed by using endovascular trapping techniques to limit collateral vessels with serial MR imaging and neurologic assessments. MATERIALS AND METHODS: Eight adult rhesus monkeys (female, 7-13 years of age) underwent MR imaging and Spetzler neurologic assessment followed by endovascular stroke induction consisting of superselective endovascular placement of surgical silk sutures into the right MCA by using a trapping technique. Two initial subjects were euthanized immediately following postocclusion MR imaging. The subsequent 6 subjects recovered and underwent follow-up MR imaging and Spetzler neurologic assessments at 48 hours, with 4 being followed to 96 hours. Stroke infarct volumes were measured, and the longitudinal Spetzler clinical neurologic scores were assessed. The brain tissues were harvested and prepared with hematoxylin-eosin staining. RESULTS: Focal permanent cerebral ischemia was induced in the targeted right MCA territory in all subjects. The volumes of the ischemic lesions at 6, 48, and 96 hours were 3.18 ± 1.007 mL (standard error of the mean) (n = 8), 6.70 ± 1.666 mL (standard error of the mean) (n = 6), and 7.23 ± 1.371 mL (standard error of the mean) (n = 4). For the survival animals, the immediate postsurgical Spetzler grading score improved from 60.7 at 24 hours to 68.7 at 48 hours. CONCLUSIONS: We report a trapping modification to an established endovascular suture stroke model that yielded reproducible ischemia and clinically quantifiable neurologic deficits with no strokes in nontarget areas. This technique may be useful in evaluating translational stroke and penumbral imaging research in addition to preclinical testing of neuroprotective therapies.
Assuntos
Modelos Animais de Doenças , Infarto da Artéria Cerebral Média/patologia , Animais , Procedimentos Endovasculares , Feminino , Macaca mulatta , Imageamento por Ressonância MagnéticaRESUMO
OBJECTIVE: To determine markers that are associated with the durability of virologic response to therapy with HIV protease inhibitors in HIV-infected individuals. DESIGN: This study encompassed two retrospective analyses of the duration of virologic response to protease inhibitor therapy. The first analysis included 29 patients receiving either monotherapy or combination therapy with the protease inhibitor ritonavir whose plasma HIV RNA levels rebounded from the point of greatest decline with mutations associated with resistance to ritonavir. The second analysis included a cohort of 102 patients who initially responded to randomized treatment with either monotherapy with ritonavir or combination therapy with ritonavir and zidovudine. METHODS: Durability of response was defined as the time from the initiation of therapy to the point at which plasma HIV RNA displayed a sustained increase of at least 0.6 log10 copies/ml from the nadir value. In the first analysis, durability of response was analyzed with respect to baseline HIV RNA, HIV RNA at the nadir, and the drop in HIV RNA from baseline to the nadir. In the second analysis, time to rebound was examined using Kaplan-Meier analysis, stratifying by either baseline HIV RNA or HIV RNA at the nadir. RESULTS: In both analyses, the durability of response was not highly associated with either baseline RNA or the magnitude of RNA decline from baseline. Instead, a strong relationship was observed between the durability of response and the nadir plasma HIV-1 RNA value (P < 0.01). The nadir in viral load was generally reached after 12 weeks of randomized therapy. CONCLUSIONS: Viral RNA determinations at intermediate timepoints may be prognostic of impending virologic failure of protease inhibitor therapy. Therapeutic strategies that allow intensification of initial antiretroviral regimens in the subset of patients with incomplete virological response before the emergence of high level resistance should be investigated.
Assuntos
Fármacos Anti-HIV/uso terapêutico , Infecções por HIV/tratamento farmacológico , Infecções por HIV/virologia , Inibidores da Protease de HIV/uso terapêutico , HIV-1/efeitos dos fármacos , Valor Preditivo dos Testes , RNA Viral/sangue , Quimioterapia Combinada , HIV-1/genética , HIV-1/fisiologia , Humanos , Mutação , Estudos Retrospectivos , Ritonavir/uso terapêutico , Resultado do Tratamento , Carga Viral , Zidovudina/uso terapêuticoRESUMO
OBJECTIVE: To evaluate the safety and antiretroviral activity of ritonavir (Norvir) and saquinavir (Invirase) combination therapy in patients with HIV infection. DESIGN: A multicenter, randomized, open-label clinical trial. SETTING: Seven HIV research units in the USA and Canada. PATIENTS: A group of 141 adults with HIV infection, CD4 T lymphocyte counts of 100-500 x 10(6) cells/l, whether treated previously or not with reverse transcriptase inhibitor therapy, but without previous HIV protease inhibitor drug therapy. INTERVENTIONS: After discontinuation of prior therapy for 2 weeks, group I patients were randomized to receive either combination (A) ritonavir 400 mg and saquinavir 400 mg twice daily or (B) ritonavir 600 mg and saquinavir 400 mg twice daily. After an initial safety assessment of group I patients, group II patients were randomized to receive either (C) ritonavir 400 mg and saquinavir 400 mg three times daily or (D) ritonavir 600 mg and saquinavir 600 mg twice daily. Investigators were allowed to add up to two reverse transcriptase inhibitors (including at least one with which the patient had not been previously treated) to a patient's regimen after week 12 for failure to achieve or maintain an HIV RNA level < or = 200 copies/ml documented on two consecutive occasions. MEASUREMENTS: Plasma HIV RNA levels and CD4+ T-lymphocyte counts were measured at baseline, every 2 weeks for 2 months, and monthly thereafter. Safety was assessed through the reporting of adverse events, physical examinations, and the monitoring of routine laboratory tests. RESULTS: The 48 weeks of study treatment was completed by 75% (106/141) of the patients. Over 80% of the patients on treatment at week 48 had an HIV RNA level < or = 200 copies/ml. In addition, intent-to-treat and on-treatment analyses revealed comparable results. Suppression of plasma HIV RNA levels was similar for all treatment arms (mean areas under the curve minus baseline through 48 weeks were-1.9, -2.0, -1.6, -1.8 log10 copies/ml in ritonavir-saquinavir 400-400 mg twice daily, 600-400 mg twice daily, 400-400 mg three times daily, and 600-600 mg twice daily, respectively). Median CD4 T-lymphocyte count rose by 128 x 10(6) cells/l from baseline, with an interquartile range (IQR) of 82-221 x 10(6) cells/l. The most common adverse events were diarrhea, circumoral paresthesia, asthenia, and nausea. Reversible elevation of serum transaminases (> 5 x upper limit of normal) occurred in 10% (14/141) of the patients enrolled in this study and was associated with baseline abnormalities in liver function tests, baseline hepatitis B surface antigen positivity, or hepatitis C antibody positivity (relative risk, 5.0; 95% confidence interval 1.5-16.9). Most moderate or severe elevations in liver function tests occurred in patients treated with ritonavir-saquinavir 600-600 mg twice daily. CONCLUSIONS: Ritonavir 400 mg combined with saquinavir 400 mg twice daily with the selective addition of reverse transcriptase inhibitors was the best-tolerated regimen of four dose-ranging regimens and was equally as active as the higher dose combinations in HIV-positive patients without previous protease inhibitor treatment.
Assuntos
Fármacos Anti-HIV/uso terapêutico , Infecções por HIV/tratamento farmacológico , Inibidores da Protease de HIV/uso terapêutico , HIV-1 , Ritonavir/uso terapêutico , Saquinavir/uso terapêutico , Adulto , Fármacos Anti-HIV/efeitos adversos , Fármacos Anti-HIV/farmacocinética , Qualidade de Produtos para o Consumidor , Quimioterapia Combinada , Feminino , Infecções por HIV/líquido cefalorraquidiano , Infecções por HIV/mortalidade , Infecções por HIV/virologia , Inibidores da Protease de HIV/efeitos adversos , Inibidores da Protease de HIV/farmacocinética , HIV-1/genética , Humanos , Masculino , Inibidores da Transcriptase Reversa/uso terapêutico , Ritonavir/efeitos adversos , Ritonavir/farmacocinética , Saquinavir/efeitos adversos , Saquinavir/farmacocinéticaRESUMO
A series of novel, pseudo-symmetrical difluoroketones which are highly potent inhibitors of the HIV-1 protease (IC50 = 1.55-0.02 nM) were synthesized. These compounds also possess good antiviral activity by inhibition of the cytopathic effect of HIV-13B in MT-4 cells in vitro.
Assuntos
Antivirais/farmacologia , Inibidores da Protease de HIV/farmacologia , Cetonas/farmacologia , Antivirais/química , Fenômenos Químicos , Físico-Química , Inibidores da Protease de HIV/química , Cetonas/química , Estrutura Molecular , Proteínas Recombinantes/antagonistas & inibidores , Relação Estrutura-AtividadeRESUMO
The design and synthesis of renin inhibitors that incorporate the novel dipeptide isostere (4S,5S)-5-amino-6-cyclohexyl-4-hydroxyhex-1-ene-2-carboxylic acid as a transition-state analogue are described. Titanium-promoted condensation of dilithiated N-alkylmethacrylamides with protected amino aldehydes results in efficient preparation of protected dipeptide analogues 7 and 8. Incorporation of 7 into the partial sequence of angiotensinogen affords potent in vitro inhibitors of human renin. Further chemical manipulation of the unsaturated amide moiety allows the study of structure-activity relationships in both the P1' and P2' sites. Details of the syntheses, stereochemical determinations, and in vitro renin inhibition are presented.
Assuntos
Dipeptídeos/síntese química , Renina/antagonistas & inibidores , Dipeptídeos/farmacologia , Humanos , Conformação Molecular , Relação Estrutura-AtividadeRESUMO
Solid phase synthesis of non-oligomeric organic compounds has been pursued for high-efficiency generation of large numbers of structurally diverse compounds for drug screening. Known as chemical diversity libraries or combinatorial libraries (when the synthesis is carried out in a combinatorial fashion), these compounds can be used for de novo discovery of drug leads or for expedient structure--activity relationship (SAR) studies. To expand the scope of solid phase synthesis beyond the capability of the traditional method of solid phase synthesis for peptides, a strategy was developed for bi-directional solid phase synthesis starting with diamino alcohol or diamino diol core structures. The strategy relies on using bifunctional linkers to modify the core structures, simultaneously protecting the hydroxyl group or the diol moiety of the core and providing a carboxyl group for attachment of the modified cores to a solid support. The two NH2 groups of the modified cores attached to the solid support were then deprotected and reacted with a wide variety of amine-reactive reagents (carboxylic acids, sulfonyl chlorides, isocyanates, chloroformates, etc.) to extend the molecule in both directions. This strategy was successfully applied to automated parallel synthesis of a library of C2 symmetric inhibitors of HIV protease containing the known symmetry-based diamino diol and diamino alcohol core structures, thus enabling expedient access of large numbers of analogs in this series. A library of over 300 discrete compounds was synthesized using this methodology in order to identify potent (IC50 < 100 nM) HIV protease inhibitors with reduced size. This paper describes the technical aspects of this technology.
Assuntos
Inibidores da Protease de HIV/síntese química , Álcoois/química , Álcoois/farmacologia , Estudos de Viabilidade , Inibidores da Protease de HIV/química , Humanos , Compostos de Metilureia/química , Piridinas/química , Relação Estrutura-Atividade , Valina/análogos & derivadosRESUMO
The synthesis of a series of renin inhibitors in which the P2 and P3 amino acids are replaced with the hydroxyethylene dipeptide isostere is reported. In vitro evaluation of the inhibitors has revealed that this isostere is an acceptable amide-bond replacement in which activity is maintained and stability is enhanced. Structure-activity relationships of this series resemble but do not parallel those of the corresponding dipeptide-containing inhibitors.
Assuntos
Dipeptídeos/farmacologia , Renina/antagonistas & inibidores , Fenômenos Químicos , Química , Cicloexanos , Dipeptídeos/síntese química , Lactonas , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
A series of low-nanomolar renin inhibitors containing novel C-terminal heterocycles has been designed by formally cyclizing the C-terminus of a glycol-based inhibitor to the second hydroxyl. Molecular modeling suggests that the heterocyclic oxygen hydrogen bonds as an acceptor to the flap region of renin and that the second hydroxyl in the glycol-based inhibitors behaves similarly.
Assuntos
Carbamatos/farmacologia , Furanos/farmacologia , Compostos Heterocíclicos , Lactonas/farmacologia , Renina/antagonistas & inibidores , Carbamatos/síntese química , Fenômenos Químicos , Química , Furanos/síntese química , Compostos Heterocíclicos/síntese química , Compostos Heterocíclicos/farmacologia , Ligação de Hidrogênio , Lactonas/síntese química , Conformação Molecular , Relação Estrutura-AtividadeRESUMO
The structure-activity relationships in two series of novel, symmetry-based inhibitors of HIV protease, the enzyme responsible for maturation of the human immunodeficiency virus, are described. Beginning with lead compounds 3-6, the effect of adding polar, heterocyclic end groups to one or both ends of the symmetric or pseudosymmetric inhibitors was probed. Aqueous solubility was enhanced > 1000-fold while maintaining potent inhibition of purified HIV-1 protease and anti-HIV activity in vitro. Pharmacokinetic studies in rats indicated a substantial difference in the absorption properties of mono-ol-based and diol-based inhibitors. The oral bioavailability of inhibitor 19 in rats was 19%; however, the Cmax obtained failed to exceed the anti-HIV EC50 in vitro. Substantial plasma levels of potent inhibitors of the diol class were not obtained after oral administration in rats; however, the optimal combination of aqueous solubility and in vitro antiviral activity of several inhibitors support their potential use in intravenous therapy.
Assuntos
Amino Álcoois/química , Diaminas/química , Inibidores da Protease de HIV/química , Amino Álcoois/farmacologia , Fenômenos Químicos , Físico-Química , Efeito Citopatogênico Viral/efeitos dos fármacos , Diaminas/farmacologia , Inibidores da Protease de HIV/farmacologia , HIV-1/efeitos dos fármacos , HIV-1/enzimologia , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
The E2 protein is required for the replication of human papillomaviruses (HPVs), which are responsible for anogenital warts and cervical carcinomas. Using an NMR-based screen, we tested compounds for binding to the DNA-binding domain of the HPV-E2 protein. Three classes of compounds were identified which bound to two distinct sites on the protein. Biphenyl and biphenyl ether compounds containing a carboxylic acid bind to a site near the DNA recognition helix and inhibit the binding of E2 to DNA. Benzophenone-containing compounds which lack a carboxylic acid group bind to the beta-barrel formed by the dimer interface and exhibit negligible effects on the binding of E2 to DNA. Structure-activity relationships from the biphenyl and biphenyl ether compounds were combined to produce a compound [5-(3'-(3",5"-dichlorophenoxy)-phenyl)-2,4-pentadienoic acid] with an IC50 value of approximately 10 microM. This compound represents a useful lead for the development of antiviral agents that interfere with HPV replication and further illustrates the usefulness of the SAR by NMR method in the drug discovery process.
Assuntos
Antivirais/química , Proteínas de Ligação a DNA/antagonistas & inibidores , DNA/metabolismo , Desenho de Fármacos , Proteínas Repressoras/metabolismo , Transativadores/metabolismo , Proteínas Virais/antagonistas & inibidores , Antivirais/farmacologia , Sítios de Ligação , Compostos de Bifenilo/farmacologia , Papillomavirus Bovino 1 , Cristalografia por Raios X , Proteínas de Ligação a DNA/metabolismo , Humanos , Ligantes , Espectroscopia de Ressonância Magnética , Modelos Químicos , Modelos Moleculares , Papillomaviridae , Conformação Proteica , Relação Estrutura-Atividade , Proteínas Virais/metabolismoRESUMO
The structure-activity studies leading to the potent and clinically efficacious HIV protease inhibitor ritonavir are described. Beginning with the moderately potent and orally bioavailable inhibitor A-80987, systematic investigation of peripheral (P3 and P2') heterocyclic groups designed to decrease the rate of hepatic metabolism provided analogues with improved pharmacokinetic properties after oral dosing in rats. Replacement of pyridyl groups with thiazoles provided increased chemical stability toward oxidation while maintaining sufficient aqueous solubility for oral absorption. Optimization of hydrophobic interactions with the HIV protease active site produced ritonavir, with excellent in vitro potency (EC50 = 0.02 microM) and high and sustained plasma concentrations after oral administration in four species. Details of the discovery and preclinical development of ritonavir are described.
Assuntos
Inibidores da Protease de HIV/química , Protease de HIV/metabolismo , Ritonavir/análogos & derivados , Ritonavir/química , Administração Oral , Animais , Disponibilidade Biológica , Cristalografia por Raios X , Protease de HIV/química , Inibidores da Protease de HIV/farmacocinética , Inibidores da Protease de HIV/farmacologia , Taxa de Depuração Metabólica , Modelos Moleculares , Conformação Molecular , Estrutura Molecular , Conformação Proteica , Piridinas/química , Piridinas/farmacocinética , Piridinas/farmacologia , Ratos , Proteínas Recombinantes/antagonistas & inibidores , Proteínas Recombinantes/química , Ritonavir/farmacocinética , Ritonavir/farmacologia , Solubilidade , Relação Estrutura-AtividadeRESUMO
The design, synthesis, and molecular modeling studies of a novel series of azacyclic ureas, which are inhibitors of human immunodeficiency virus type 1 (HIV-1) protease that incorporate different ligands for the S1', S2, and S2' substrate-binding sites of HIV-1 protease are described. The synthesis of this series is highly flexible in the sense that the P1', P2, and P2' residues of the inhibitors can be changed independently. Molecular modeling studies on the phenyl ring of the P2 and P2' ligand suggested incorporation of hydrogen-bonding donor/acceptor groups at the 3' and 4-positions of the phenyl ring should increase binding potency. This led to the discovery of compound 7f (A-98881), which possesses high potency in the HIV-1 protease inhibition assay and the in vitro MT-4 cell culture assay (Ki = approximately 5 pM and EC50 = 0.002 microM). This compares well with the symmetrical cyclic urea 1 pioneered at DuPont Merck.
Assuntos
Inibidores da Protease de HIV/síntese química , Inibidores da Protease de HIV/farmacologia , HIV-1/enzimologia , Sítios de Ligação , Resistência Microbiana a Medicamentos , Protease de HIV/metabolismo , HIV-1/efeitos dos fármacos , Modelos MolecularesRESUMO
A C2 symmetry-based HIV protease inhibitor, A77003, exerts potent antiviral activity against a wide spectrum of HIV isolates in vitro. In this study, we asked whether A77003 could cause irreversible conformational changes to HIV-1, whether the amounts of viral RNA and p24 capsid protein per virion were altered, and how the infectivity of the virus produced in the presence of the drug was affected. We found that the number of viral particles and per-virion viral RNA content of the virus produced in the presence of A77003 did not significantly differ from those of the virus produced in the absence of the drug, whereas significant morphological changes were observed as assessed by transmission electron microscopy. However, the virus produced in the presence of A77003 contained substantially less p24gag protein per virion particle as compared to those produced in the absence of the drug or in the presence of AZT. Virions produced in the presence of A77003 showed up to 50-fold less infectious capability in subsequent tissue culture than control virions produced in the absence of drug or in the presence of AZT. This reduction in infectivity was maintained for at least 10 days in culture. The present data suggest that A77003 impairs HIV-1 protease-mediated Gag processing, interferes with the assembly and maturation of the virus, and leads to an irreversible loss of the infectivity of the virus, although a low but positive level of reversion to infectivity during the 10-day assay occurs. These features of A77003 (and perhaps similar HIV protease inhibitors as well) anti-HIV activity should represent desirable properties for antiviral therapy of AIDS and related diseases.
Assuntos
Antivirais/farmacologia , Infecções por HIV/prevenção & controle , Inibidores da Protease de HIV/farmacologia , HIV-1/efeitos dos fármacos , Compostos de Metilureia , Piridinas , Sequência de Bases , Linhagem Celular , Produtos do Gene gag/análise , Infecções por HIV/tratamento farmacológico , HIV-1/patogenicidade , HIV-1/ultraestrutura , Dados de Sequência Molecular , RNA Viral/análise , Valina/análogos & derivados , Vírion/efeitos dos fármacos , Zidovudina/farmacologiaAssuntos
Inibidores da Protease de HIV/química , Protease de HIV/química , HIV-1/enzimologia , Peptídeos/química , Sequência de Aminoácidos , Sítios de Ligação , Desenho de Fármacos , Protease de HIV/metabolismo , Inibidores da Protease de HIV/metabolismo , Inibidores da Protease de HIV/farmacologia , HIV-1/efeitos dos fármacos , Ligação de Hidrogênio , Dados de Sequência Molecular , Peptídeos/metabolismo , Peptídeos/farmacologia , Ligação Proteica , Conformação Proteica , Relação Estrutura-Atividade , Especificidade por SubstratoAssuntos
Fármacos Anti-HIV/farmacologia , Inibidores da Protease de HIV/farmacologia , HIV/efeitos dos fármacos , Síndrome da Imunodeficiência Adquirida/tratamento farmacológico , Síndrome da Imunodeficiência Adquirida/metabolismo , Síndrome da Imunodeficiência Adquirida/virologia , Animais , Fármacos Anti-HIV/metabolismo , Fármacos Anti-HIV/uso terapêutico , Citocromo P-450 CYP3A , Sistema Enzimático do Citocromo P-450/metabolismo , Resistência Microbiana a Medicamentos/genética , Quimioterapia Combinada , Genótipo , HIV/genética , Inibidores da Protease de HIV/metabolismo , Inibidores da Protease de HIV/uso terapêutico , Humanos , Oxigenases de Função Mista/metabolismo , Mutação/efeitos dos fármacos , Fenótipo , RNA Viral/sangue , Ratos , Carga ViralRESUMO
A new stereocontrolled synthesis of the psi[CH = CH] dipeptide isostere is described. They key step of the sequence relies on the stereospecific alpha-alkylation of delta-amino-gamma-mesyloxy-alpha,beta-unsaturated esters. The broad availability of nucleophilic alpha-side chains by this method allows the preparation of a wide variety of psi[CH = CH] isosteres with predictable stereochemistry.
Assuntos
Dipeptídeos/química , Estereoisomerismo , Alquilação , Ésteres/química , Modelos MolecularesRESUMO
The structural proteins and enzymes of the human immunodeficiency virus type 1 core are translated as part of two polyprotein precursors, Gag and Gag-Pol, which are cleaved by a virally encoded protease. Viruses grown in the presence of inhibitors of the protease contain core particles that are aberrantly assembled, and upon infection of susceptible cells, they do not synthesize viral DNA. Through the use of a proteinase inhibitor (A77003), we determined that the viral reverse transcriptase can efficiently synthesize viral DNA as part of the unprocessed Gag-Pol precursor. We also found that the stabilities of core particles composed of unprocessed precursors were considerably enhanced. These observations suggest that for viruses composed of unprocessed precursors, replication is interrupted before the reverse transcription step.