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Experimental and Toxicologic Pathology... Nov 2007Nucleoside analogues are used in the treatment of viral infections, including those caused by human immunodeficiency virus, cytomegalovirus, and herpes virus. These...
Nucleoside analogues are used in the treatment of viral infections, including those caused by human immunodeficiency virus, cytomegalovirus, and herpes virus. These drugs are beneficial in the treatment of human disease, but are associated with toxicities that often limit their intended therapeutic use, including anemia, neutropenia, peripheral neuropathy, and myopathy. Some of these compounds have been reported to be carcinogenic in rodents. To investigate the carcinogenic potential of lobucavir, a nucleoside analogue, three groups of 60 male and female mice were orally administered lobucavir at daily doses of 10, 50, and 250 mg/kg (males) or 30, 150, and 750 mg/kg (females) over a period of 104 weeks. Two identical groups of 60 male and female mice each served as controls. The morphology and the incidence of neoplasms is described and compared with the tumor spectrum of other nucleoside analogues. Light microscopically, lobucavir-induced neoplastic lesions consisted of upper digestive tract squamous cell neoplasia in males and females; cervical, vaginal, and cutaneous squamous cell neoplasia in females; and Hardarian gland adenomas and adenocarcinomas in male mice. These results suggest that long-term administration of lobucavir causes neoplasia in mice, the spectrum of which resembles that observed after long-term administration of zidovudine or ganciclovir.
Topics: Administration, Oral; Animals; Animals, Outbred Strains; Anti-HIV Agents; Carcinogenicity Tests; Carcinogens; Carcinoma, Squamous Cell; Dose-Response Relationship, Drug; Female; Guanine; Hyperplasia; Longevity; Male; Mice; Neoplasms, Experimental
PubMed: 17942294
DOI: 10.1016/j.etp.2007.09.002 -
Antimicrobial Agents and Chemotherapy Dec 1998The guanosine analogs BMS-200475 and lobucavir have previously been shown to effectively suppress propagation of the human hepatitis B virus (HBV) and woodchuck...
The guanosine analogs BMS-200475 and lobucavir have previously been shown to effectively suppress propagation of the human hepatitis B virus (HBV) and woodchuck hepatitis virus (WHV) in 2.2.15 liver cells and in the woodchuck animal model system, respectively. This repression was presumed to occur via inhibition of the viral polymerase (Pol) by the triphosphate (TP) forms of BMS-200475 and lobucavir which are both produced in mammalian cells. To determine the exact mode of action, BMS-200475-TP and lobucavir-TP, along with several other guanosine analog-TPs and lamivudine-TP were tested against the HBV, WHV, and duck hepatitis B virus (DHBV) polymerases in vitro. Estimates of the 50% inhibitory concentrations revealed that BMS-200475-TP and lobucavir-TP inhibited HBV, WHV, and DHBV Pol comparably and were superior to the other nucleoside-TPs tested. More importantly, both analogs blocked the three distinct phases of hepadnaviral replication: priming, reverse transcription, and DNA-dependent DNA synthesis. These data suggest that the modest potency of lobucavir in 2.2.15 cells may be the result of poor phosphorylation in vivo. Kinetic studies revealed that BMS-200475-TP and lobucavir-TP competitively inhibit HBV Pol and WHV Pol with respect to the natural dGTP substrate and that both drugs appear to bind to Pol with very high affinities. Endogenous sequencing reactions conducted in replicative HBV nucleocapsids suggested that BMS-200475-TP and lobucavir-TP are nonobligate chain terminators that stall Pol at sites that are distinct yet characteristically two to three residues downstream from dG incorporation sites.
Topics: Antiviral Agents; DNA-Directed RNA Polymerases; Deoxyguanosine; Enzyme Inhibitors; Guanine; Hepadnaviridae; Hepatitis B Virus, Duck; Hepatitis B Virus, Woodchuck; Hepatitis B virus; Humans; Kinetics; Nucleic Acid Synthesis Inhibitors; Phosphates; RNA-Directed DNA Polymerase; Structure-Activity Relationship
PubMed: 9835515
DOI: 10.1128/AAC.42.12.3200 -
Antimicrobial Agents and Chemotherapy Dec 1997Lobucavir (LBV) is a deoxyguanine nucleoside analog with broad-spectrum antiviral activity. LBV was previously shown to inhibit herpes simplex virus (HSV) DNA polymerase...
Lobucavir (LBV) is a deoxyguanine nucleoside analog with broad-spectrum antiviral activity. LBV was previously shown to inhibit herpes simplex virus (HSV) DNA polymerase after phosphorylation by the HSV thymidine kinase. Here we determined the mechanism of action of LBV against human cytomegalovirus (HCMV). LBV inhibited HCMV DNA synthesis to a degree comparable to that of ganciclovir (GCV), a drug known to target the viral DNA polymerase. The expression of late proteins and RNA, dependent on viral DNA synthesis, was also inhibited by LBV. Immediate-early and early HCMV gene expression was unaffected, suggesting that LBV acts temporally coincident with HCMV DNA synthesis and not through cytotoxicity. In vitro, the triphosphate of LBV was a potent inhibitor of HCMV DNA polymerase with a Ki of 5 nM. LBV was phosphorylated to its triphosphate form intracellularly in both infected and uninfected cells, with phosphorylated metabolite levels two- to threefold higher in infected cells. GCV-resistant HCMV isolates, with deficient GCV phosphorylation due to mutations in the UL97 protein kinase, remained sensitive to LBV. Overall, these results suggest that LBV-triphosphate halts HCMV DNA replication by inhibiting the viral DNA polymerase and that LBV phosphorylation can occur in the absence of viral factors including the UL97 protein kinase. Furthermore, LBV may be effective in the treatment of GCV-resistant HCMV.
Topics: Antiviral Agents; Cells, Cultured; Cytomegalovirus; Cytomegalovirus Infections; DNA Replication; DNA, Viral; Drug Resistance, Microbial; Enzyme Inhibitors; Fibroblasts; Ganciclovir; Guanine; Humans; Nucleic Acid Synthesis Inhibitors; Phosphorylation; Viral Proteins
PubMed: 9420038
DOI: 10.1128/AAC.41.12.2680 -
Antiviral Research Dec 2000Lobucavir (BMS-180194), a cyclobutyl-guanosine nucleoside analogue, effectively reduced WHV-viremia in chronically infected carrier woodchucks (Marmota monax) by daily...
Antiviral efficacy of lobucavir (BMS-180194), a cyclobutyl-guanosine nucleoside analogue, in the woodchuck (Marmota monax) model of chronic hepatitis B virus (HBV) infection.
Lobucavir (BMS-180194), a cyclobutyl-guanosine nucleoside analogue, effectively reduced WHV-viremia in chronically infected carrier woodchucks (Marmota monax) by daily per os treatment. WHV-viremia in the animals was measured by the serum content of hybridizable WHV-genomic DNA. Lobucavir, given at daily doses of 10 and 20 mg/kg body weight, reduced WHV-viremia by a 10- to 200-fold range during therapy. Lobucavir, given at 5 mg/kg, suppressed WHV-viremia by a 10- to 30-fold range, whereas a 0.5 mg/kg dose had no significant effect. WHV-viremia was also measured by hepadnaviral endogenous polymerase activity (EPA) in sera of animals treated for 6 weeks at 5 and 0.5 mg/kg. Changes in EPA in sera of lobucavir treated animals were comparable to changes in WHV DNA levels. Viremia in treated carriers recrudesced to pretreatment levels by 2 weeks of therapy cessation. These results indicated that the minimally effective antiviral daily per os dose of lobucavir in WHV-carrier woodchucks was approximately 5 mg/kg.
Topics: Animals; DNA, Viral; Disease Models, Animal; Guanine; Hepatitis B Virus, Woodchuck; Hepatitis B, Chronic; Humans; Marmota; Reverse Transcriptase Inhibitors; Viremia
PubMed: 11164506
DOI: 10.1016/s0166-3542(00)00128-5 -
Bioorganic & Medicinal Chemistry Dec 2000Synthesis of lobucavir prodrug, L-valine, [(1S,2R,3R)-3-(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl)-2-(hydroxymethyl) cyclobutyl]methyl ester monohydrochloride (BMS...
Synthesis of lobucavir prodrug, L-valine, [(1S,2R,3R)-3-(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl)-2-(hydroxymethyl) cyclobutyl]methyl ester monohydrochloride (BMS 233866), requires regioselective coupling of one of the two hydroxyl groups of lobucavir (BMS 180194) with valine. Either hydroxyl group of lobucavir could be selectively aminoacylated with valine by using enzymatic reactions. N-[(Phenylmethoxy)carbonyl]-L-valine, [(1R,2R,4S)-2-(2-amino-6-oxo-1H-purin-9-yl)-4-(hydroxymethyl)cyclobutyl]methyl ester (3, 82.5% yield), was obtained by selective hydrolysis of N,N'-bis[(phenylmethoxy)carbonyl]bis[L-valine], O,O'-[(1S,2R,3R)-3-(2-amino-6-oxo-1H-purin-9-yl)cyclobuta-1,2-diyl]methyl ester (1) with lipase M, and L-valine, [(1R,2R,4S)-2-(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl)-4-(hydroxymethyl)cyclobutyl]methyl ester monohydrochloride (4, 87% yield) was obtained by hydrolysis of bis[L-valine], O,O'-[(1S,2R,3R)-3-(2-amino-6-oxo-1H-purin-9-yl)cyclobuta-1,2-diyl]methyl ester, dihydrochloride (2), with lipase from Candida cylindracea. The final intermediate for lobucavir prodrug, N-[(phenylmethoxy)carbonyl]-L-valine, [(1S,2R,4R)-3-(2-amino-6-oxo-1H-purin-9-yl)-2-(hydroxymethyl)cyclobutyl]methyl ester (5), could be obtained by transesterification of lobucavir using ChiroCLEC BL (61% yield), or more selectively by using immobilized lipase from Pseudomonas cepacia (84% yield).
Topics: Acylation; Burkholderia cepacia; Guanine; Hydrolysis; In Vitro Techniques; Lipase; Prodrugs
PubMed: 11131158
DOI: 10.1016/s0968-0896(00)00209-1 -
Transplantation Mar 1999Mycophenolate mofetil (MMF) has been approved as an immunosuppressive agent in kidney transplant recipients and may thus be used concomitantly with antiherpetic agents,...
The immunosuppressive agent mycophenolate mofetil markedly potentiates the activity of lobucavir [1R(1alpha,2beta,3alpha)]-9-[2,3-bis(hydroxymethyl)cyclobutyl]guanine against different herpes viruses.
BACKGROUND
Mycophenolate mofetil (MMF) has been approved as an immunosuppressive agent in kidney transplant recipients and may thus be used concomitantly with antiherpetic agents, the latter for the treatment of intercurrent herpesvirus infections. The parent compound of MMF, mycophenolic acid (MPA), is a potent inhibitor of inosine monophosphate dehydrogenase and causes depletion of the intracellular (deoxy)guanosine triphosphate [(d)GTP] pools. Lobucavir [1R(1alpha,2beta,3alpha)]-9-[2,3-bis(hydroxymethyl)cyc lobutyl]guanine (LBV) is a novel antiviral agent with activity against ganciclovir-resistant cytomegalovirus (CMV) strains, that is in phase II clinical trials for the treatment of CMV infections. LBV triphosphate inhibits the viral DNA polymerase competitively with dGTP. We present the results of our studies on the antiviral effects of the combinations LBV + MMF and LBV + MPA.
METHODS
The antiviral effects of LBV either alone or in combination with MMF or MPA on the replication of CMV, herpes simplex virus type- (HSV) 1 (HSV-1), HSV-2, and thymidine kinase-deficient HSV-1 were studied by means of plaque or CPE reduction assays.
RESULTS
When combined with LBV, MPA (at concentrations ranging from 0.25 to 10 microg/ml, which are readily attainable in human plasma) markedly potentiated the antiviral efficacy of LBV against HSV-1 and HSV-2, that is a 10- to 100-fold decrease in EC50. Moreover, the EC50 of LBV against TK- HSV-1 decreased up to 1400-fold upon combination with MPA. MPA by itself had little or no effect on the replication of these viruses. Moreover, MPA and MMF resulted in a marked increase in the anti-CMV activity of LBV minimal FIC (FICmin: 0.24 and 0.26, respectively). Exogenously added guanosine reversed the potentiating effect of MPA on the antiviral activity of LBV, which indicates that this potentiating effect results from a depletion of the endogenous dGTP pools, thus favoring the inhibitory action of the LBV-triphosphate on the viral DNA polymerase. Ribavirin, another inhibitor of inosine monophosphate-dehydrogenase, also caused a marked enhancement of the antiviral activity of LBV against HSV-1 (12-fold), HSV-2 (20-fold), and TK- HSV-1 (25-fold).
CONCLUSION
MMF markedly potentiates the activity of LBV against HSV-1, HSV-2, TK- HSV-1, and CMV. This drug interaction may have important implications when using LBV in the treatment of intercurrent herpesvirus infections in transplant recipients under MMF therapy.
Topics: Animals; Anti-HIV Agents; Cells, Cultured; Chlorocebus aethiops; Drug Synergism; Guanine; Herpes Simplex; Herpesvirus 1, Human; Humans; Immunosuppressive Agents; Mycophenolic Acid; Vero Cells; Virus Replication
PubMed: 10096537
DOI: 10.1097/00007890-199903150-00022 -
Drugs Jul 1996Of the large number of agents under development for the treatment of herpes virus infections [herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), varicella zoster... (Review)
Review
Of the large number of agents under development for the treatment of herpes virus infections [herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), varicella zoster virus (VZV), cytomegalovirus (CMV), Epstein-Barr virus (EBV)], only ten have apparently reached clinical development. Aciclovir was approved for the treatment of HSV infections over 10 years ago, and it remains an important and reliable antiviral agent. Recent approvals in some countries of valaciclovir for VZV infection and famciclovir for both HSV and VZV infections demonstrate the rapidity of change in this field. Intravenous ganciclovir and foscarnet are approved for the treatment of CMV infection in the immunocompromised patient. Five of the antiherpetic drugs under current clinical development are nucleoside analogues or their prodrugs; another is a phosphorylated nucleoside (nucleotide). Four of the nucleoside agents-penciclovir, famciclovir, valaciclovir and lobucavir-are being developed for the management of HSV and VZV infections. Valaciclovir is also being developed for the prevention of CMV infections and famciclovir and lobucavir for the treatment of hepatitis B virus infection. Oral ganciclovir, lobucavir, ISIS 2922 and cidofovir are being developed for the suppression of CMV infections in immunocompromised patients. Sorivudine has been studied in VZV infections. n-Docosanol is under development for HSV infections, and cidofovir is being developed for both HSV and CMV infections, as well as for treatment of other viral diseases. Traditionally, the adverse effects associated with anti-CMV compounds have been more difficult to manage and are acceptable clinically only because of the severity of the underlying infection and lack of safer therapeutic alternatives. In general, toxicity issues continue to be problematic in the anti-CMV arena, although newer agents have improved the situation to some extent. In contrast, the safety of anti-HSV compounds has traditionally been excellent, establishing a safety standard that must be met by newer agents entering the field.
Topics: Antiviral Agents; Biological Availability; Clinical Trials as Topic; Herpesviridae Infections; Humans; Prodrugs
PubMed: 8799682
DOI: 10.2165/00003495-199652010-00002 -
Journal of Viral Hepatitis Mar 2000Several nucleoside analogues (penciclovir, lobucavir, dioxalane guanine [DXG], 1-beta-2,6-diaminopurine dioxalane [DAPD], L-FMAU, lamivudine) and acyclic nucleoside...
Several nucleoside analogues (penciclovir, lobucavir, dioxalane guanine [DXG], 1-beta-2,6-diaminopurine dioxalane [DAPD], L-FMAU, lamivudine) and acyclic nucleoside phosphonate analogues (adefovir, tenofovir) that are in clinical use, in clinical trials or under preclinical development for the treatment of hepatitis B virus (HBV) infections, were evaluated for their inhibitory effect on the replication of a la- mivudine-resistant HBV variant containing the methionine --> valine substitution (M550V) in the polymerase nucleoside-binding domain. The antiviral activity was determined in the tetracycline-responsive HepAD38 and HepAD79 cells, which are stably transfected with either a cDNA copy of the wild-type pregenomic RNA or with cDNA containing the M550V mutation. As expected, lamivudine was much less ( approximately 200-fold) effective at inhibiting replication of the M550V mutant virus than the wild-type virus. In contrast, adefovir, tenofovir, lobucavir, L-FMAU, DXG and DAPD proved almost equally effective against both viruses. A second objective of this study was to directly compare the antiviral potency of the anti-HBV agents in HepG2 2.2.15 cells (which are routinely used for anti-HBV drug-screening purposes) with that in HepAD38 cells. HepAD38 cells produce much larger quantities of HBV than HepG2 2.2.15 cells, and thus allow drug screening in a multiwell plate format. All compounds were found to be almost equally effective at inhibiting HBV replication in HepAD38 cells (as in HepG2 2.2.15 cells), except for penciclovir, which was clearly less effective in HepAD38 cells.
Topics: Acyclovir; Adenine; Antiviral Agents; Arabinofuranosylcytosine Triphosphate; Cell Line; DNA, Viral; DNA-Directed DNA Polymerase; Dioxolanes; Guanine; Hepatitis B virus; Humans; Organophosphonates; Organophosphorus Compounds; Purine Nucleosides; Tenofovir; Virus Replication
PubMed: 10760047
DOI: 10.1046/j.1365-2893.2000.00210.x -
Journal of Pharmaceutical Sciences Oct 2006The regional absorption of lobucavir (LBV), an experimental antiviral agent, and ganciclovir (DHPG) was investigated in rabbit intestine using an in situ single-pass...
The regional absorption of lobucavir (LBV), an experimental antiviral agent, and ganciclovir (DHPG) was investigated in rabbit intestine using an in situ single-pass perfusion technique. Duodenal, jejunal, and colonic segments in anesthetized rabbits were perfused with drug solutions in a hypotonic buffer at 0.2 mL/min. Effluent perfusate samples for drug analysis were collected every 10 min for 180 min. To account for water absorption during perfusion, an intestinal absorption model was developed to estimate the absorptive clearance (PeA): PeA=Qavexln((QinxCin)/(QoutxCout)), where Qave is a logarithmic average of the inflow (Qin) and outflow perfusion rate (Qout); Cin and C(out) are drug inflow and outflow concentrations. The PeA of LBV in the duodenum and jejunum was 2.1+/-0.77 and 1.7+/-0.46 microL/min/cm (n=3), respectively, 4.8- and 3.0-fold higher than that of DHPG in the same animals. However, LBV PeA decreased significantly in the colon (0.47+/-0.11 microL/min/cm) and was similar to that of DHPG which exhibited no regional differences in absorption. The interplay between PeA and solubility was studied using a compartmental absorption and transit model, and simulations were performed to investigate dose-limited absorption and the sources of variability in absorption where two compounds differ significantly. The dose range where absorption started to decrease was predicted using the model, with LBV exhibiting the phenomenon at a lower dose than DHPG (450 vs. 750 mg). Furthermore, the intersubject variability in human absorption of both compounds was reproduced when the variability in both PeA and the small intestinal transit time was considered in the model. The variability in the ascending colonic transit time also contributed to the intersubject variability observed for DHPG. The results demonstrate value of integrating in situ studies and modeling in predicting these absorption characteristics.
Topics: Animals; Antiviral Agents; Computer Simulation; Ganciclovir; Gastric Mucosa; Guanine; Humans; Intestinal Absorption; Intestinal Mucosa; Male; Models, Biological; Rabbits; Water
PubMed: 16883564
DOI: 10.1002/jps.20716 -
Current Opinion in Hematology Nov 1998Cytomegalovirus (CMV) infection remains a major cause of morbidity and mortality in recipients of an allogeneic stem cell transplant (SCT). Due to the broad application... (Review)
Review
Cytomegalovirus (CMV) infection remains a major cause of morbidity and mortality in recipients of an allogeneic stem cell transplant (SCT). Due to the broad application of antiviral prophylaxis and preemptive therapy, a decrease in early-onset and a subsequent increase in late-onset CMV disease has been observed. New data on the latency state and reactivation of CMV have been presented, the role of T-cell responses in the control of CMV further substantiated, and viral immune escape mechanisms described in more detail. Sensitive diagnostic assays using nucleic acid amplification and hybridization techniques have been commercialized and will allow standardization of CMV diagnostics in antiviral drug trials. Quantification of the viral load will be increasingly considered for initiation and, in patients with persistence of high viral titers despite antiviral therapy, screening for antiviral drug resistance. Clinical data are emerging to show that, apart from ganciclovir, foscarnet can be given safely even after allogeneic SCT. Additional drugs like lobucavir and cidofovir have been used for specific indications. Interactions of immunosuppressive drugs and antiviral compounds of clinical relevance have been described. Thus, therapeutic drug monitoring and targeted antiviral drug dosing will become standard practice in antiviral treatment strategies in patients following allogeneic SCT.
Topics: Antiviral Agents; Cytomegalovirus Infections; Drug Interactions; Hematopoietic Stem Cell Transplantation; Humans; Immunosuppressive Agents; Opportunistic Infections; Viral Load
PubMed: 9814660
DOI: 10.1097/00062752-199811000-00022