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1.
Chem Biodivers ; 18(1): e2000937, 2021 Jan.
Article in English | MEDLINE | ID: mdl-33270983

ABSTRACT

Challenges resulting from novel viruses or new strains of known viruses call for new antiviral agents. Nucleoside analogs that act as inhibitors of viral polymerases are an attractive class of antivirals. For nucleosides containing thymine, base pairing is weak, making it desirable to identify nucleobase analogs that pair more strongly with adenine, in order to compete successfully with the natural substrate. We have recently described a new class of strongly binding thymidine analogs that contain an ethynylmethylpyridone as base and a C-nucleosidic linkage to the deoxyribose. Here we report the synthesis of the 3'-azido-2',3'-deoxyribose derivative of this compound, dubbed AZW, both as free nucleoside and as ProTide phosphoramidate. As a proof of principle, we studied the activity against Herpes simplex virus type 1 (HSV1). Whereas the ProTide phosphoramidate suffered from low solubility, the free nucleoside showed a stronger inhibitory effect than that of AZT in a plaque reduction assay. This suggests that strongly pairing C-nucleoside analogs of pyrimidines have the potential to become active pharmaceutical ingredients with antiviral activity.


Subject(s)
Antiviral Agents/pharmacology , Herpesvirus 1, Human/drug effects , Nucleosides/chemistry , Zidovudine/analogs & derivatives , Amides/chemistry , Animals , Antiviral Agents/chemical synthesis , Antiviral Agents/chemistry , Chlorocebus aethiops , Humans , Phosphoric Acids/chemistry , Prodrugs/chemical synthesis , Prodrugs/chemistry , Prodrugs/pharmacology , Pyridines/chemistry , Vero Cells , Zidovudine/pharmacology
2.
Bioorg Med Chem Lett ; 30(17): 127365, 2020 09 01.
Article in English | MEDLINE | ID: mdl-32738968

ABSTRACT

The present study aimed to investigate the effect of AZT derivates containing tellurium (Te) on human breast cancer cell lines and the mechanisms underlying cell death. The inhibitory effect of AZT and its derivatives (7m and 7r) was determined by the MTT assay (6.25, 12.5, 25, 50 and 100 µM in 24 and 48 h time points), meanwhile the induction of apoptosis and the cell cycle phases was investigated by flow cytometry. The MTT assay showed that AZT derivatives decreased the rate of cell proliferation at concentrations of 12.5 µM, while commercial AZT showed low antitumor potential. In flow cytometric analysis, we demonstrate that the AZT derivatives do not induce apoptosis at the concentration tested and promote the cell cycle arrest in the S phase. Besides, predicted absorption, distribution, metabolization, excretion and toxicity analysis suggest that the compounds possess a good pharmacokinetic profile and possibly less toxicity when compared to conventional AZT. These compounds containing tellurium in their formulation are potential therapeutic agents for breast cancer.


Subject(s)
Antineoplastic Agents/chemical synthesis , Zidovudine/analogs & derivatives , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacokinetics , Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Cell Line, Tumor , Cell Survival/drug effects , Female , Half-Life , Humans , S Phase Cell Cycle Checkpoints/drug effects , Tellurium/chemistry , Triple Negative Breast Neoplasms/metabolism , Triple Negative Breast Neoplasms/pathology , Zidovudine/chemical synthesis , Zidovudine/pharmacokinetics , Zidovudine/pharmacology
3.
Bioorg Chem ; 95: 103499, 2020 01.
Article in English | MEDLINE | ID: mdl-31838287

ABSTRACT

Phosphazine and phosphazide derivatives are described herein as a new class of selective and potent acetylcholinesterase (AChE) inhibitors and ß-amyloid aggregation inhibitors. Phosphazines (5-7) were synthesized smoothly via a redox-condensation reaction of 1,2-bis(diphenylphosphino)ethane with different amines derivatives in the presence of dialkyl azodicarboxylate (Staudinger reaction) while phosphazides (8) via electrophilic attack of azido derivatives. Structures of the synthesized compounds were justified on the basis of compatible elementary and spectroscopic analyses. All the compounds were evaluated for their acetylcholinesterase inhibitory activity. The most three potent compounds (5b-c and 8b) showing AChE IC50 values (29.85-34.96 nM) comparable to that of donepezil (34.42 nM) were subjected to further investigation by testing their butyrylcholinesterase, MMP-2 and self-induced Aß aggregation inhibition activity. Especially, the coumarin phosphazide derivative (8b) presented the best AChE inhibition selectivity index (IC50 = 34.96 nM, AChE/BuChE; 3.81) together with good inhibition ability against MMP-2 (IC50 = 441.33 nM) and self-induced Aß1-42 aggregation (IC50 = 337.77 nM). In addition, the inhibition of metal-induced Aß aggregation by 8b was confirmed by thioflavine T fluorescence. The most potent effect of 8b was observed on the Zn2+-induced Aß42 aggregation. Kinetic study of compound 8b suggested it to be a competitive AChE inhibitor. Also, it specifically chelates metal and is predicted to be permeable to BBB. It also possesses low toxicity on SH-SY5Y neuroblastoma cells with a safety index of 15.37. In addition, it was demonstrated that compound 8b can improve the cognitive impairment of scopolamine-induced model in mice with % alternations and transfer latency time comparable to that of donepezil. Also, a docking study was carried out and it was in accordance with the in vitro results. These promising in vitro and in vivo findings highlight compound 8b as a possible drug candidate in searching for new multifunctional AD drugs.


Subject(s)
Acetylcholinesterase/metabolism , Alzheimer Disease/drug therapy , Amyloid beta-Peptides/antagonists & inhibitors , Aziridines/pharmacology , Cholinesterase Inhibitors/pharmacology , Organophosphonates/pharmacology , Zidovudine/analogs & derivatives , Alzheimer Disease/metabolism , Amnesia/chemically induced , Amnesia/drug therapy , Amnesia/metabolism , Amyloid beta-Peptides/metabolism , Animals , Aziridines/chemical synthesis , Aziridines/chemistry , Cholinesterase Inhibitors/chemical synthesis , Cholinesterase Inhibitors/chemistry , Dose-Response Relationship, Drug , Humans , Kinetics , Ligands , Maze Learning/drug effects , Mice , Models, Molecular , Molecular Structure , Organophosphonates/chemical synthesis , Organophosphonates/chemistry , Protein Aggregates/drug effects , Scopolamine , Structure-Activity Relationship , Zidovudine/chemical synthesis , Zidovudine/chemistry , Zidovudine/pharmacology
4.
Chembiochem ; 19(18): 1939-1943, 2018 09 17.
Article in English | MEDLINE | ID: mdl-29953711

ABSTRACT

Metabolic incorporation of bioorthogonal functional groups into cellular nucleic acids can be impeded by insufficient phosphorylation of nucleosides. Previous studies found that 5azidomethyl-2'-deoxyuridine (AmdU) was incorporated into the DNA of HeLa cells expressing a low-fidelity thymidine kinase, but not by wild-type HeLa cells. Here we report that membrane-permeable phosphotriester derivatives of AmdU can exhibit enhanced incorporation into the DNA of wild-type cells and animals. AmdU monophosphate derivatives bearing either 5'-bispivaloyloxymethyl (POM), 5'-bis-(4-acetoxybenzyl) (AB), or "Protide" protective groups were used to mask the phosphate group of AmdU prior to its entry into cells. The POM derivative "POM-AmdU" exhibited better chemical stability, greater metabolic incorporation efficiency, and lower toxicity than "AB-AmdU". Remarkably, the addition of POM-AmdU to the water of zebrafish larvae enabled the biosynthesis of azide-modified DNA throughout the body.


Subject(s)
Azides/chemistry , DNA/chemistry , Nucleotides/chemistry , Zidovudine/analogs & derivatives , Animals , Azides/metabolism , Cell Membrane Permeability , Click Chemistry , DNA/metabolism , Esters/chemistry , Esters/metabolism , HeLa Cells , Humans , Nucleotides/metabolism , Zebrafish , Zidovudine/chemistry , Zidovudine/metabolism
5.
Anal Bioanal Chem ; 410(21): 5245-5253, 2018 Aug.
Article in English | MEDLINE | ID: mdl-29947896

ABSTRACT

Asymmetric flow field-flow fractionation (AF4) coupled with UV-Vis spectroscopy, multi-angle light scattering (MALS) and refractive index (RI) detection has been applied for the characterization of MIL-100(Fe) nanoMOFs (metal-organic frameworks) loaded with nucleoside reverse transcriptase inhibitor (NRTI) drugs for the first time. Empty nanoMOFs and nanoMOFs loaded with azidothymidine derivatives with three different degrees of phosphorylation were examined: azidothymidine (AZT, native drug), azidothymidine monophosphate (AZT-MP), and azidothymidine triphosphate (AZT-TP). The particle size distribution and the stability of the nanoparticles when interacting with drugs have been determined in a time frame of 24 h. Main achievements include detection of aggregate formation in an early stage and monitoring nanoMOF morphological changes as indicators of their interaction with guest molecules. AF4-MALS proved to be a useful methodology to analyze nanoparticles engineered for drug delivery applications and gave fundamental data on their size distribution and stability. Graphical abstract ᅟ.


Subject(s)
Anti-HIV Agents/administration & dosage , Coordination Complexes/chemistry , Drug Carriers/chemistry , Metal-Organic Frameworks/chemistry , Nanoparticles/chemistry , Zidovudine/administration & dosage , Anti-HIV Agents/chemistry , Antimetabolites/administration & dosage , Antimetabolites/chemistry , Dideoxynucleotides/administration & dosage , Dideoxynucleotides/chemistry , Dynamic Light Scattering , Fractionation, Field Flow , Models, Molecular , Particle Size , Refractometry , Spectrophotometry, Ultraviolet , Thymine Nucleotides/administration & dosage , Thymine Nucleotides/chemistry , Zidovudine/analogs & derivatives , Zidovudine/chemistry
6.
Nucleic Acids Res ; 44(5): 2310-22, 2016 Mar 18.
Article in English | MEDLINE | ID: mdl-26850643

ABSTRACT

We analyzed a multi-drug resistant (MR) HIV-1 reverse transcriptase (RT), subcloned from a patient-derived subtype CRF02_AG, harboring 45 amino acid exchanges, amongst them four thymidine analog mutations (TAMs) relevant for high-level AZT (azidothymidine) resistance by AZTMP excision (M41L, D67N, T215Y, K219E) as well as four substitutions of the AZTTP discrimination pathway (A62V, V75I, F116Y and Q151M). In addition, K65R, known to antagonize AZTMP excision in HIV-1 subtype B was present. Although MR-RT harbored the most significant amino acid exchanges T215Y and Q151M of each pathway, it exclusively used AZTTP discrimination, indicating that the two mechanisms are mutually exclusive and that the Q151M pathway is obviously preferred since it confers resistance to most nucleoside inhibitors. A derivative was created, additionally harboring the TAM K70R and the reversions M151Q as well as R65K since K65R antagonizes excision. MR-R65K-K70R-M151Q was competent of AZTMP excision, whereas other combinations thereof with only one or two exchanges still promoted discrimination. To tackle the multi-drug resistance problem, we tested if the MR-RTs could still be inhibited by RNase H inhibitors. All MR-RTs exhibited similar sensitivity toward RNase H inhibitors belonging to different inhibitor classes, indicating the importance of developing RNase H inhibitors further as anti-HIV drugs.


Subject(s)
Drug Resistance, Multiple, Viral/genetics , Enzyme Inhibitors/pharmacology , HIV Reverse Transcriptase/antagonists & inhibitors , HIV-1/drug effects , Ribonuclease H, Human Immunodeficiency Virus/antagonists & inhibitors , Amino Acid Sequence , Amino Acid Substitution , Anti-HIV Agents/chemistry , Anti-HIV Agents/pharmacology , Cloning, Molecular , Dideoxynucleotides/chemistry , Dideoxynucleotides/pharmacology , Enzyme Inhibitors/chemistry , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Expression , Genotype , HIV Infections/drug therapy , HIV Infections/virology , HIV Reverse Transcriptase/genetics , HIV Reverse Transcriptase/metabolism , HIV-1/enzymology , HIV-1/genetics , HIV-1/isolation & purification , Humans , Molecular Sequence Data , Mutagenesis, Site-Directed , Mutation , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Ribonuclease H, Human Immunodeficiency Virus/genetics , Ribonuclease H, Human Immunodeficiency Virus/metabolism , Thymine Nucleotides/chemistry , Thymine Nucleotides/pharmacology , Zidovudine/analogs & derivatives , Zidovudine/chemistry , Zidovudine/pharmacology
7.
Biochemistry ; 56(1): 33-46, 2017 Jan 10.
Article in English | MEDLINE | ID: mdl-27936595

ABSTRACT

Reverse transcriptases (RTs) are typically assayed in vitro with 5-10 mM Mg2+, whereas the free Mg2+ concentration in cells is much lower. Artificially high Mg2+ concentrations used in vitro can misrepresent different properties of human immunodeficiency virus (HIV) RT, including fidelity, catalysis, pausing, and RNase H activity. Here, we analyzed nucleoside (NRTIs) and non-nucleoside RT inhibitors (NNRTIs) in primer extension assays at different concentrations of free Mg2+. At low concentrations of Mg2+, NRTIs and dideoxynucleotides (AZTTP, ddCTP, ddGTP, and 3TCTP) inhibited HIV-1 and HIV-2 RT synthesis less efficiently than they did with large amounts of Mg2+, whereas inhibition by the "translocation-defective RT inhibitor" EFdA (4'-ethynyl-2-fluoro-2'-deoxyadenosine) was unaffected by Mg2+ concentrations. Steady-state kinetic analyses revealed that the reduced level of inhibition at low Mg2+ concentrations resulted from a 3-9-fold (depending on the particular nucleotide and inhibitor) less efficient incorporation (based on kcat/Km) of these NRTIs under this condition compared to incorporation of natural dNTPs. In contrast, EFdATP was incorporated with an efficiency similar to that of its analogue dATP at low Mg2+ concentrations. Unlike NRTIs, NNRTIs (nevirapine, efavirenz, and rilviripine), were approximately 4-fold (based on IC50 values) more effective at low than at high Mg2+ concentrations. Drug-resistant HIV-1 RT mutants also displayed the Mg2+-dependent difference in susceptibility to NRTIs and NNRTIs. In summary, analyzing the efficiency of inhibitors under more physiologically relevant low-Mg2+ conditions yielded results dramatically different from those from measurements using commonly employed high-Mg2+ in vitro conditions. These results also emphasize differences in Mg2+ sensitivity between the translocation inhibitor EFdATP and other NRTIs.


Subject(s)
Dideoxynucleotides/pharmacology , HIV Reverse Transcriptase/antagonists & inhibitors , Magnesium/pharmacology , Nucleosides/pharmacology , Reverse Transcriptase Inhibitors/pharmacology , Deoxycytosine Nucleotides/pharmacology , Deoxyguanine Nucleotides/pharmacology , Dose-Response Relationship, Drug , Drug Interactions , Electrophoresis, Polyacrylamide Gel , HIV Reverse Transcriptase/genetics , HIV Reverse Transcriptase/metabolism , Humans , Kinetics , Mutation , Thymine Nucleotides/pharmacology , Zalcitabine/pharmacology , Zidovudine/analogs & derivatives , Zidovudine/pharmacology
8.
Bioorg Med Chem Lett ; 27(8): 1627-1632, 2017 04 15.
Article in English | MEDLINE | ID: mdl-28285913

ABSTRACT

The development of prodrugs has progressed with the aim of improving drug bioavailability by overcoming various barriers that reduce drug benefits in clinical use, such as stability, duration, water solubility, side effect profile, and taste. Many conventional drugs act as the precursors of an active agent in vivo; for example, the anti-HIV agent azidothymidine (AZT) is converted into its corresponding active triphosphate ester in the body, meaning that AZT is a prodrug in the broadest sense. However prodrug design is generally difficult owing to the lack of general versatility. Thus, these prodrugs, broadly defined, are often discovered by chance or trial-and-error. Recently, many prodrugs that could release the corresponding parent drugs with or without enzymatic action under physiological conditions have been reported. These prodrugs can be easily designed and synthesized because of their generally applicable modifications. This digest paper provides an overview of recent development in prodrug strategies for drugs with a carboxylic acid or hydroxyl/amino group on the basis of a generally applicable modification strategy, such as esterification, amidation, or benzylation.


Subject(s)
Anti-HIV Agents/chemistry , Anti-HIV Agents/pharmacokinetics , Drug Design , Prodrugs/chemistry , Prodrugs/pharmacokinetics , Amides/chemistry , Amides/metabolism , Amides/pharmacokinetics , Animals , Anti-HIV Agents/metabolism , Benzyl Compounds/chemistry , Benzyl Compounds/metabolism , Benzyl Compounds/pharmacokinetics , Biological Availability , Carboxylic Acids/chemistry , Carboxylic Acids/metabolism , Carboxylic Acids/pharmacokinetics , Esterification , HIV/drug effects , HIV Infections/drug therapy , Humans , Prodrugs/metabolism , Solubility , Zidovudine/analogs & derivatives , Zidovudine/metabolism , Zidovudine/pharmacokinetics
9.
Bioorg Med Chem ; 25(19): 5128-5132, 2017 10 01.
Article in English | MEDLINE | ID: mdl-28712845

ABSTRACT

The brain provides a sanctuary site for HIV due, in part, to poor penetration of antiretroviral agents at the blood-brain barrier. This lack of penetration is partially attributed to drug efflux transporters such as P-glycoprotein (P-gp) and ABCG2. Inhibition of both ABCG2 and P-gp is critical for enhancing drug accumulation into the brain. In this work, we have developed a class of homodimers based on the HIV reverse transcriptase inhibitor azidothymidine (AZT) that effectively inhibits P-gp and ABCG2. These agents block transporter mediated efflux of the P-gp substrate calcein-AM and the ABCG2 substrate mitoxantrone. The homodimers function by interacting with the transporter drug binding sites as demonstrated by competition studies with the photo-affinity agent and P-gp/ABCG2 substrate [125I]iodoarylazidoprazosin. As such, these dual inhibitors of both efflux transporters provide a model for the future development of delivery vehicles for antiretroviral agents to the brain.


Subject(s)
ATP Binding Cassette Transporter, Subfamily B, Member 1/antagonists & inhibitors , ATP Binding Cassette Transporter, Subfamily G, Member 2/antagonists & inhibitors , Biological Transport/drug effects , Blood-Brain Barrier/drug effects , Neoplasm Proteins/antagonists & inhibitors , Zidovudine/analogs & derivatives , Zidovudine/pharmacology , ATP Binding Cassette Transporter, Subfamily B, Member 1/metabolism , ATP Binding Cassette Transporter, Subfamily G, Member 2/metabolism , Antiviral Agents/chemistry , Antiviral Agents/pharmacology , Blood-Brain Barrier/metabolism , Cell Line , Dimerization , Humans , Neoplasm Proteins/metabolism
10.
Vopr Virusol ; 62(1): 5-11, 2017.
Article in English | MEDLINE | ID: mdl-29323840

ABSTRACT

Federation Convincing evidence for high therapeutic activity and tolerability of Phosphazide in the treatment of HIV/AIDS-infection is given. Phosphazide is currently used in various regimens of highly active antiretroviral therapy, as well as in the HIV therapy in patients with simultaneously acquired chronic hepatitis C or tuberculosis. Therapeutic possibilities of Phosphazide were clearly manifested in the prevention of HIV transmission from mother to child. There is every reason to use Phosphazide in first-line antiretroviral therapy.


Subject(s)
Anti-HIV Agents/therapeutic use , HIV Infections/drug therapy , HIV/drug effects , Infectious Disease Transmission, Vertical/prevention & control , Organophosphonates/therapeutic use , Pregnancy Complications, Infectious/drug therapy , Zidovudine/analogs & derivatives , Anti-HIV Agents/pharmacokinetics , Antiretroviral Therapy, Highly Active/methods , Female , Fetus , HIV/genetics , HIV/metabolism , HIV Infections/transmission , HIV Infections/virology , Humans , Organophosphonates/pharmacokinetics , Pregnancy , Pregnancy Complications, Infectious/virology , Treatment Outcome , Zidovudine/pharmacokinetics , Zidovudine/therapeutic use
11.
Vopr Virusol ; 61(1): 34-9, 2016.
Article in Russian | MEDLINE | ID: mdl-27145599

ABSTRACT

Efficacy, safety, and economical aspects of treatment with abacavir, zidovudine, stavudine, and phosphazide in the schemes of antiretroviral therapy of the HIV-infected patients receiving therapy for hepatitis C virus were tested. Clinical, immunological, and virologic efficacy of treatment and dynamics of hemoglobin, thrombocytes, and alanine aminotransferase as markers of common adverse events recorded at the start of the antiviral therapy of chronic hepatitis C and after 4, 8, 12, 24, 48 weeks of the treatment were evaluated. The usage of these drugs in the schemes of antiretroviral therapy exhibited efficacy, high tolerability and safety for all HIV reverse transcriptase inhibitors.


Subject(s)
Dideoxynucleosides/therapeutic use , HIV Infections/drug therapy , Hepatitis C, Chronic/drug therapy , Organophosphonates/therapeutic use , Reverse Transcriptase Inhibitors/therapeutic use , Stavudine/therapeutic use , Zidovudine/analogs & derivatives , Zidovudine/therapeutic use , Adult , Alanine Transaminase/blood , Biomarkers/blood , Blood Platelets/drug effects , CD4-Positive T-Lymphocytes/drug effects , Cell Count , Coinfection , Dideoxynucleosides/economics , Female , Genotype , HIV Infections/blood , HIV Infections/virology , HIV-1/drug effects , HIV-1/genetics , HIV-1/growth & development , Hemoglobins/metabolism , Hepacivirus/drug effects , Hepacivirus/genetics , Hepacivirus/growth & development , Hepatitis C, Chronic/blood , Hepatitis C, Chronic/virology , Humans , Male , Organophosphonates/economics , Reverse Transcriptase Inhibitors/economics , Stavudine/economics , Zidovudine/economics
12.
Klin Med (Mosk) ; 94(4): 295-299, 2016.
Article in English | MEDLINE | ID: mdl-28957610

ABSTRACT

AIM: Comparative assessment of the efficiency of application of different therapeutic schemes for post-contact prevention (PCP) of HIV infection in health providers. METHODS: Medical personnel that had professional contacts with HIV-infected patients (n=44) were given medications for PCP. 19 of them (group 1) used phosphazide, 25 (group 2) combivir (lamivudine + zidovudine) in combination with kaletra for 4 weeks after the contact. Phosphazide (AZT Farma K.B., Russia) was used at a dose of 0.4 g twice daily, other medications in standard doses. The results were evaluated 4 weeks, 3, 6, and 12 months after PCP from the safety of the treatment and the absence of professional HIV infection. RESULTS: The medical personnel showed no signs of HIV infection throughout the entire period of observation. The safety of therapy was confirmed by the absence of myelohepatotoxic effect of the preparations. Combivir therapy caused a 1.8-fold rise in AST activity of within 4 weeks after onset of PCP (p<0,05). Phosphazide produced no such effect. CONCLUSION: The above results indicate that both schemes ofantiretroviral activity are 100% efficient as PCP of HIV infection, but phosphazide has an advantage of higher safety and better tolerability.


Subject(s)
HIV Infections , Infectious Disease Transmission, Patient-to-Professional/prevention & control , Lamivudine/administration & dosage , Lopinavir/administration & dosage , Organophosphonates/administration & dosage , Post-Exposure Prophylaxis/methods , Ritonavir/administration & dosage , Zidovudine/analogs & derivatives , Adult , Anti-HIV Agents/administration & dosage , Anti-HIV Agents/adverse effects , Drug Combinations , Drug Monitoring/methods , Female , HIV Infections/prevention & control , HIV Infections/transmission , Humans , Lopinavir/adverse effects , Male , Organophosphonates/adverse effects , Ritonavir/adverse effects , Russia , Treatment Outcome , Zidovudine/administration & dosage , Zidovudine/adverse effects
13.
Antimicrob Agents Chemother ; 59(10): 6395-401, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26239974

ABSTRACT

The male genital tract is a potential site of viral persistence. Therefore, adequate concentrations of antiretrovirals are required to eliminate HIV replication in the genital tract. Despite higher zidovudine (ZDV) and lamivudine (3TC) concentrations in seminal plasma (SP) than in blood plasma (BP) (SP/BP drug concentration ratios of 2.3 and 6.7, respectively), we have previously reported lower relative intracellular concentrations of their active metabolites, zidovudine triphosphate (ZDV-TP) and lamivudine triphosphate (3TC-TP), in seminal mononuclear cells (SMCs) than in peripheral blood mononuclear cells (PBMCs) (SMC/PBMC drug concentration ratios of 0.36 and 1.0, respectively). Here, we use population pharmacokinetic (PK) modeling-based methods to simultaneously describe parent and intracellular metabolite PK in blood, semen, and PBMCs and SMCs. From this model, the time to steady state in each matrix was estimated, and the results indicate that the PK of 3TC-TP and ZDV-TP in PBMCs are different from the PK of the two in SMCs and different for the two triphosphates. We found that steady-state conditions in PBMCs were achieved within 2 days for ZDV-TP and 3 days for 3TC-TP. However, steady-state conditions in SMCs were achieved within 2 days for ZDV-TP and 2 weeks for 3TC-TP. Despite this, or perhaps because of it, ZDV-TP in SMCs does not achieve the surrogate 50% inhibitory concentration (IC50) (as established for PBMCs, assuming SMC IC50 = PBMC IC50) at the standard 300-mg twice-daily dosing. Mechanistic studies are needed to understand these differences and to explore intracellular metabolite behavior in SMCs for other nucleoside analogues used in HIV prevention, treatment, and cure.


Subject(s)
Anti-HIV Agents/pharmacokinetics , Cytidine Triphosphate/analogs & derivatives , Dideoxynucleotides/pharmacokinetics , Lamivudine/analogs & derivatives , Leukocytes, Mononuclear/metabolism , Models, Statistical , Semen/metabolism , Thymine Nucleotides/pharmacokinetics , Zidovudine/analogs & derivatives , Adult , Anti-HIV Agents/pharmacology , Biological Availability , Biological Transport , Blood Cells/drug effects , Blood Cells/metabolism , Blood Cells/pathology , Blood Cells/virology , Computer Simulation , Cytidine Triphosphate/pharmacokinetics , Cytidine Triphosphate/pharmacology , Dideoxynucleotides/pharmacology , HIV Infections/drug therapy , HIV Infections/pathology , HIV Infections/virology , HIV-1/drug effects , HIV-1/physiology , Humans , Lamivudine/pharmacokinetics , Lamivudine/pharmacology , Leukocytes, Mononuclear/drug effects , Leukocytes, Mononuclear/pathology , Leukocytes, Mononuclear/virology , Male , Semen/cytology , Semen/drug effects , Semen/virology , Thymine Nucleotides/pharmacology , Time Factors , Zidovudine/pharmacokinetics , Zidovudine/pharmacology
14.
Bioorg Med Chem ; 23(9): 2168-75, 2015 May 01.
Article in English | MEDLINE | ID: mdl-25801161

ABSTRACT

A system for delivery of analogues of AZT-triphosphates (AZT*TP) based on SiO2 nanoparticles was proposed. For this purpose, a simple and versatile method was developed for the preparation of SiO2∼dNTP conjugates using the 'click'-reaction between AZTTP and premodified nanoparticles containing the alkyne groups. The substrate properties of SiO2∼AZT*TP were tested using Klenow fragment and HIV reverse transcriptase. The 3'-triazole derivatives of thymidine triphosphate being a part of the SiO2∼AZT*TP nanocomposites were shown to be incorporated into the growing DNA chain. It was shown by confocal microscopy that the proposed SiO2∼AZT*TP nanocomposites penetrate into cells. These nanocomposites were shown to inhibit the reproduction of POX and Herpes viruses at nontoxic concentrations.


Subject(s)
Dideoxynucleotides/administration & dosage , Dideoxynucleotides/chemistry , Drug Delivery Systems , Nanoparticles/chemistry , Silicon Dioxide/chemistry , Simplexvirus/drug effects , Thymine Nucleotides/administration & dosage , Thymine Nucleotides/chemistry , Triazoles/chemistry , Variola virus/drug effects , Zidovudine/analogs & derivatives , Animals , Cell Survival/drug effects , Cells, Cultured , Chlorocebus aethiops , Click Chemistry , Dideoxynucleotides/pharmacology , Dose-Response Relationship, Drug , Microbial Sensitivity Tests , Molecular Structure , Simplexvirus/growth & development , Structure-Activity Relationship , Thymine Nucleotides/pharmacology , Variola virus/growth & development , Vero Cells , Zidovudine/administration & dosage , Zidovudine/chemistry , Zidovudine/pharmacology
15.
J Labelled Comp Radiopharm ; 57(8): 540-9, 2014 Jun 30.
Article in English | MEDLINE | ID: mdl-24992010

ABSTRACT

The nucleosides zidovudine (AZT), stavudine (d4T), and telbivudine (LdT) are approved for use in the treatment of human immunodeficiency virus (HIV) and hepatitis B virus (HBV) infections. To promote positron emission tomography (PET) imaging studies on their pharmacokinetics, pharmacodynamics, and applications in cancer diagnosis, a convenient one-pot method for Pd(0)-Cu(I) co-mediated rapid C-C coupling of [(11)C]methyl iodide with stannyl precursor was successfully established and applied to synthesize the PET tracers [(11)C]zidovudine, [(11)C]stavudine, and [(11)C]telbivudine. After HPLC purification and radiopharmaceutical formulation, the desired PET tracers were obtained with high radioactivity (6.4-7.0 GBq) and specific radioactivity (74-147 GBq/µmol) and with high chemical (>99%) and radiochemical (>99.5%) purities. This one-pot Pd(0)-Cu(I) co-mediated rapid C-[(11)C]methylation also worked well for syntheses of [methyl-(11)C]thymidine and [methyl-(11)C]4'-thiothymidine, resulting twice the radioactivity of those prepared by a previous two-pot method. The mechanism of one-pot Pd(0)-Cu(I) co-mediated rapid C-[(11)C]methylation was also discussed.


Subject(s)
Radiopharmaceuticals/chemical synthesis , Zidovudine/chemical synthesis , Carbon Radioisotopes/chemistry , Catalysis , Chemistry Techniques, Synthetic/methods , Copper/chemistry , Methylation , Palladium/chemistry , Zidovudine/analogs & derivatives
16.
J Chromatogr Sci ; 62(5): 399-405, 2024 May 31.
Article in English | MEDLINE | ID: mdl-38520317

ABSTRACT

A novel method has been developed for the estimation of N-Nitroso dimethyl amine impurities (NDMA) and N-Nitroso diethyl amine (NDEA) in Zidovudine by using Gas chromatograph Triple Quadrupole Mass with Liquid autosampler (GC-MS/MS) and the method is validated as per International Conference on Harmonization recommendations. Sample analysis was executed for Zidovudine by developed method. Both NDMA and NDEA were detected in below quantitation limit for the Zidovudine batches. Efficient chromatographic separation was achieved on a DB-WAX 30 m length × 0.25 mm internal diameter, 0.5-µm film thickness, Triple quad-8040 GC-MS/MS. Quantification was carried out at Triple quad electron ionization source was at a column flow of 1.5 mL/min at a column oven temperature 50°C. The precision was in the range of 0.9-2.5% for NDMA and 0.8-2.3% for NDEA, and regression analysis shows as r value (correlation coefficient) of is >0.99. This method is capable to detect the NDMA and NDEA impurities in Zidovudine at a level of 0.006 ppm for limit of detection and 0.018 ppm for limit of quantitation with respect to test concentration of 66.66 mg/mL.


Subject(s)
Drug Contamination , Gas Chromatography-Mass Spectrometry , Limit of Detection , Tandem Mass Spectrometry , Zidovudine , Gas Chromatography-Mass Spectrometry/methods , Zidovudine/analysis , Zidovudine/chemistry , Zidovudine/analogs & derivatives , Tandem Mass Spectrometry/methods , Reproducibility of Results , Linear Models
17.
J Virol ; 86(9): 5122-33, 2012 May.
Article in English | MEDLINE | ID: mdl-22379084

ABSTRACT

Resistance to nucleoside reverse transcriptase (RT) inhibitors is conferred on human immunodeficiency virus type 1 through thymidine analogue resistance mutations (TAMs) that increase the ability of RT to excise chain-terminating nucleotides after they have been incorporated. The RT mutation M184V is a potent suppressor of TAMs. In RT containing TAMs, the addition of M184V suppressed the excision of 3'-deoxy-3'-azidothymidine monophosphate (AZTMP) to a greater extent on an RNA template than on a DNA template with the same sequence. The catalytically inactive RNase H mutation E478Q abolished this difference. The reduction in excision activity was similar with either ATP or pyrophosphate as the acceptor substrate. Decreased excision of AZTMP was associated with increased cleavage of the RNA template at position -7 relative to the primer terminus, which led to increased primer-template dissociation. Whether M184V was present or not, RT did not initially bind at the -7 cleavage site. Cleavage at the initial site was followed by RT dissociation and rebinding at the -7 cleavage site, and the dissociation and rebinding were enhanced when the M184V mutation was present. In contrast to the effect of M184V, the K65R mutation suppressed the excision activity of RT to the same extent on either an RNA or a DNA template and did not alter the RNase H cleavage pattern. Based on these results, we propose that enhanced RNase H cleavage near the primer terminus plays a role in M184V suppression of AZT resistance, while K65R suppression occurs through a different mechanism.


Subject(s)
HIV Reverse Transcriptase/genetics , HIV Reverse Transcriptase/metabolism , HIV-1/genetics , HIV-1/metabolism , Mutation , Nucleotides/metabolism , Adenosine Triphosphate/metabolism , DNA Primers/metabolism , Dideoxynucleotides/metabolism , Drug Resistance, Viral/genetics , Humans , RNA, Viral/biosynthesis , Ribonuclease H/metabolism , Thymine Nucleotides/metabolism , Zidovudine/analogs & derivatives , Zidovudine/metabolism
18.
Biomacromolecules ; 14(3): 737-42, 2013 Mar 11.
Article in English | MEDLINE | ID: mdl-23351139

ABSTRACT

The natural nucleotide adenosine triphosphate (ATP) and nucleotide analogues such as azidothymidine triphosphate (AZT-TP) display important pharmacological activities for the treatment of ischemia and HIV infections, respectively. Their clinical use is, however, limited mostly due to their hydrophilicity, which highly restricts their diffusion into the target cells. Few nanocarriers have been proposed to address the challenge of ATP/AZT-TP cellular delivery, but the loading efficiency, preparation complexity, and efficient cellular delivery remain important barriers to their development. In this study, we propose an original, straightforward and versatile design of nucleotide and nucleotide analogue nanocarriers based on the natural polysaccharide chitosan (CS). We show that the drugs ATP and AZT-TP can induce ionotropic gelation of CS, leading to CS/ATP and CS/AZT-TP nanoparticles with high drug entrapment efficiency and loading rate-up to 44%. Such nanocarriers release ATP and AZT-TP in physiological media and allow an efficient in vitro cellular delivery of these molecules down to the cell cytoplasm.


Subject(s)
Adenosine Triphosphate/pharmacology , Dideoxynucleotides/pharmacology , Drug Carriers/chemistry , Drug Delivery Systems/methods , Nanoparticles/chemistry , Nucleotides/chemistry , Zidovudine/analogs & derivatives , Adenosine Triphosphate/chemistry , Animals , Cell Survival , Chitosan/chemistry , Dideoxynucleotides/chemistry , HIV Infections/drug therapy , Macrophages/metabolism , Mice , Zidovudine/chemistry , Zidovudine/pharmacology
19.
PLoS Comput Biol ; 8(1): e1002359, 2012 Jan.
Article in English | MEDLINE | ID: mdl-22275860

ABSTRACT

Nucleoside analogs (NAs) are used to treat numerous viral infections and cancer. They compete with endogenous nucleotides (dNTP/NTP) for incorporation into nascent DNA/RNA and inhibit replication by preventing subsequent primer extension. To date, an integrated mathematical model that could allow the analysis of their mechanism of action, of the various resistance mechanisms, and their effect on viral fitness is still lacking. We present the first mechanistic mathematical model of polymerase inhibition by NAs that takes into account the reversibility of polymerase inhibition. Analytical solutions for the model point out the cellular- and kinetic aspects of inhibition. Our model correctly predicts for HIV-1 that resistance against nucleoside analog reverse transcriptase inhibitors (NRTIs) can be conferred by decreasing their incorporation rate, increasing their excision rate, or decreasing their affinity for the polymerase enzyme. For all analyzed NRTIs and their combinations, model-predicted macroscopic parameters (efficacy, fitness and toxicity) were consistent with observations. NRTI efficacy was found to greatly vary between distinct target cells. Surprisingly, target cells with low dNTP/NTP levels may not confer hyper-susceptibility to inhibition, whereas cells with high dNTP/NTP contents are likely to confer natural resistance. Our model also allows quantification of the selective advantage of mutations by integrating their effects on viral fitness and drug susceptibility. For zidovudine triphosphate (AZT-TP), we predict that this selective advantage, as well as the minimal concentration required to select thymidine-associated mutations (TAMs) are highly cell-dependent. The developed model allows studying various resistance mechanisms, inherent fitness effects, selection forces and epistasis based on microscopic kinetic data. It can readily be embedded in extended models of the complete HIV-1 reverse transcription process, or analogous processes in other viruses and help to guide drug development and improve our understanding of the mechanisms of resistance development during treatment.


Subject(s)
Anti-HIV Agents/pharmacology , HIV-1/drug effects , Nucleosides/pharmacology , Reverse Transcriptase Inhibitors/pharmacology , pol Gene Products, Human Immunodeficiency Virus/antagonists & inhibitors , Base Sequence , DNA, Viral/chemistry , DNA, Viral/metabolism , Dideoxynucleotides/pharmacology , Drug Resistance, Viral , HIV Infections/drug therapy , HIV Infections/virology , HIV Reverse Transcriptase/genetics , HIV Reverse Transcriptase/metabolism , HIV-1/enzymology , HIV-1/genetics , Humans , Kinetics , Models, Molecular , Molecular Sequence Data , Mutation , RNA, Viral/metabolism , Thymine Nucleotides/pharmacology , Zidovudine/analogs & derivatives , Zidovudine/pharmacology
20.
J Org Chem ; 78(12): 5832-42, 2013 Jun 21.
Article in English | MEDLINE | ID: mdl-23713491

ABSTRACT

Additions of lactams, imides, (S)-4-benzyl-1,3-oxazolidin-2-one, 2-pyridone, pyrimidine-2,4-diones (AZT derivatives), or inosines to the electron-deficient triple bonds of methyl propynoate, tert-butyl propynoate, 3-butyn-2-one, N-propynoylmorpholine, or N-methoxy-N-methylpropynamide in the presence of many potential catalysts were examined. DABCO and, second, DMAP appeared to be the best (highest reaction rates and E/Z ratios), while RuCl3, RuClCp*(PPh3)2, AuCl, AuCl(PPh3), CuI, and Cu2(OTf)2 were incapable of catalyzing such additions. The groups incorporated (for example, the 2-(methoxycarbonyl)ethenyl group that we name MocVinyl) serve as protecting groups for the above-mentioned heterocyclic CONH or CONHCO moieties. Deprotections were accomplished via exchange with good nucleophiles: the 1-dodecanethiolate anion turned out to be the most general and efficient reagent, but in some particular cases other nucleophiles also worked (e.g., MocVinyl-inosines can be cleaved with succinimide anion). Some structural and mechanistic details have been accounted for with the help of DFT and MP2 calculations.


Subject(s)
Imides/chemistry , Lactams/chemistry , Nucleosides/chemistry , Alkynes/chemistry , Catalysis , Electrons , Inosine/chemistry , Molecular Structure , Morpholines/chemistry , Piperazines/chemistry , Propionates/chemistry , Pyridines/chemistry , Pyridones/chemistry , Pyrimidines/chemistry , Quantum Theory , Stereoisomerism , Zidovudine/analogs & derivatives , Zidovudine/chemistry
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