A series of modified oligonucleotides containing P=S backbone and a six-membered azasugar (6-AZS) were synthesized and tested for their ability to inhibit human immunodeficiency virus (HIV) in vitro without the aid of any transfecting agents. While P=S oligonucleotides with natural nucleotides had little anti-HIV-1 activity, six-membered azasugar nucleotide (6-AZN)-containing P=S oligonucleotides (AZPSON) potently inhibited the HIV-1/SHIV production and syncytium formation in vitro (EC50 = 0.02~0.2 micro M) without cytotoxicity up to 100 micro M. AZPSONs are enzymatically stable over 6 days in culture supernatant. Phosphodiester (P=O) backbone only or mixed backbone (P=O and P=S) oligonucleotides that contain 6-AZN did not exhibit anti-HIV-1 activity. The anti-HIV-1 capacity of AZPSON seems to depend on the number and/or distribution patterns of 6-AZN in the oligonucleotides. The oligomer 2198, most effective for anti-HIV-1 activity among the AZPSONs, was much more effective than ddI or ddC in anti-HIV activity. Particularly noteworthy is that the anti-HIV-1 activity of AZPSON-2198 was better than AZT in the long-lasting efficacy after a single treatment.
Giant Cells ; HIV ; HIV-1 ; Nucleotides ; Oligonucleotides ; Phosphorothioate Oligonucleotides*

To search the effective antisense oligonucleotide that inhibit the growth of Herpes simplex virus type 1 (HSV-1), six kinds of phosphorothioate oligodeoxynucleotides (S-ODNs) were synthesized and the antiviral activity was assessed by measuring cytopathic effect in Vero cells infected with HSV-1. Of the three dodecamer S-ODNs, cornplernentary to the translation initiation site of IE2 (AS2) and scrambled S-ODN (AS1) showed more significant antiherpetic activity than AS4 complementary to the IE4. Accordingly, the antiviral effect of dodecamer S-ODN was not specific. In contrast to the no inhibitory effect of sense strand S-ODN of ICP8 (AS6), two S-ODNs complementary to the translation initiation site of ICP8 (AS3) and that of IE1 (AS5) showed potent antiherpetic activity assessed in vitro HSV-1 virus yield assay. Antiherpetic effect of AS3 was decreased in proportion to the addition of AS6. The synthesis of viral protein ICPS and IE1 were inhibited in AS3 and AS5 treated HSV-1 infected Vero cells, respectively. These findings suggest that antiherpetic effect of AS3 is specifically mediated by targeting ICPS. S-ODNs had no effect on Vero cell viability. The data suggest that the 19-mer S-ODNs may be effective in antiviral chemotherapy.
Drug Therapy ; Herpes Simplex* ; Herpesvirus 1, Human ; Oligodeoxyribonucleotides ; Phosphorothioate Oligonucleotides* ; Simplexvirus* ; Vero Cells

OBJECTIVETo determine whether 3'phosphorothioate-modified-2 terminal mismatched primers can turn off DNA polymerization mediated by Exo(+) polymerase.

METHODSTwo-directional primer extension was performed using polymerase with and without 3' exonuclease activity. The effects of unmodified primers and 3' phosphorothioate-modified primers on primer extension were evaluated.

RESULTSExo(-) polymerase yielded products from matched and mismatched primers regardless of their modification. However, 3' phosphorothioate-modified primers with a single base mismatch at -2 position worked similarly to the terminal (-1) mismatched primers in triggering the novelly reported "off-switch" of Exo(+) polymerase.

CONCLUSIONThese data suggested that the "off-switch" can be of enormous application in the diagnosis of single gene diseases and in the association studies by single nucleotide polymorphism screening.

DNA Primers ; chemistry ; genetics ; Exonucleases ; metabolism ; Humans ; Phosphorothioate Oligonucleotides ; chemistry ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide

Base Sequence ; Hepacivirus ; drug effects ; genetics ; Hepatitis C ; drug therapy ; virology ; Humans ; Molecular Sequence Data ; Oligonucleotides ; therapeutic use ; Phosphorothioate Oligonucleotides ; therapeutic use

To study the pharmacokinetics of cantide, an antisense oligonucleotide, and its metabolites after iv gtt administration in rhesus monkeys, a dual solid phase extraction pretreatment method coupling with non-gel sieving capillary electrophoresis analysis method was used for determination of cantide and its metabolites in plasma and their pharmacokinetic parameters were calculated. The pharmacokinetic behavior of cantide and its metabolites (M1 and M2) after iv gtt administration (8, 16 and 24 mg kg(-1)) in rhesus monkeys were investigated. After iv gtt administration of cantide to rhesus monkeys, cantide in plasma was eliminated rapidly and the terminal elimination half-life (t1/2) was 57.91-77.97 min, the correlation coefficients (r) to the dose of Cmax AUC(o-inf) and AUC(0-t) of the prototype was 0.9918, 0.9568 and 0.9773, respectively. The metabolites of cantide reached the Cmax following cantide immediately and the Cmax of metabolites were lower than that of the prototype. The CL(S) of cantide and its metabolites (M1 and M2) were 1.60-2.19, 5.92-8.58 and 6.07-8.78 mL min(-1) kg(-1), respectively. So, it is concluded that the Cmax of cantide and its metabolites increased with the dose, which is the same as their AUC(0-inf) and AUC(0-t). The CL(S) of metabolites were higher than that of the prototype. The MRT and t1/2 of metabolites in the high dose group increased obviously.
Animals ; Area Under Curve ; Electrophoresis, Capillary ; methods ; Female ; Half-Life ; Infusions, Intravenous ; Macaca mulatta ; Male ; Oligonucleotides, Antisense ; blood ; metabolism ; pharmacokinetics ; Phosphorothioate Oligonucleotides ; blood ; metabolism ; pharmacokinetics ; Solid Phase Extraction

Novel biologic agents that selectively target specific molecules and pathways have been developed recently for the management of inflammatory bowel disease (IBD). Anti-TNF-alpha, an antibody to TNF-alpha is one of the first newly developed drugs to dramatically improve the symptoms of patients with IBD. Therapy with anti-TNF-alpha demonstrates a new paradigm for management of IBD and early treatment with these drugs has demonstrated increased benefit. However, more than one-third of the patients have lost response to the drug. Also, there is the problem of antibody formation. Therefore, enormous efforts to develop novel drugs as an alternatives to anti-TNF-alpha are underway. Anti CD4+ T cell cytokine including IL-12/23 and IL-17 blockers, selective anti-adhesion molecules known as natalizumab, vedolizumab and alicaforsen, T-cell proliferation inhibitors, anti-inflammatory cytokines, immune stimulators, growth factors and mitogen activated protein kinase (MAPK) inhibitors are among the novel therapeutic agents that are currently being investigated for efficacy and safety in the management of IBD. The aim of this paper is to review current knowledge concerning the mechanism of action, the short and long term efficacy, and the safety of these novel biologic therapies, as well as that of anti-TNF-alpha, in IBD.
Antibodies, Monoclonal, Humanized ; Antibody Formation ; Biological Agents ; Biological Therapy ; Cytokines ; Humans ; Inflammatory Bowel Diseases ; Intercellular Signaling Peptides and Proteins ; Interleukin-17 ; Phosphorothioate Oligonucleotides ; Protein Kinases ; T-Lymphocytes ; Tumor Necrosis Factor-alpha ; Natalizumab

CpG-Oligodeoxynucleotide (ODN) has two backbones. Phosphorothioate backbone (PS) shows a strong immunostimulating effect while phosphodiester (PE) shows little in vivo. 3' hexameric deoxyriboguanosine-run (3' dG6-run) conjugation to PE CpG-ODN has been reported to enhance immunostimulation and to protect against asthma when injected at the time of sensitization in mice. We evaluated the treatment effects of PE and PS CpG-ODN with or without 3' dG6-run on asthma in presensitized mice. BALB/c mice sensitized with ovalbumin and alum were challenged with 1% ovalbumin on three days. CpG-ODNs (100 microgram) or PBS were injected 4 times; 27 hr before challenge and 3 hr before each challenge (CpG-dG6: CpG-ODN with 3' dG6-run, PE*-CpG-dG6: PE-CpG-dG6 with two PS backbones at the 5' terminus). PE-CpG showed no treatment effect. PE-CpG-dG6 only increased ovalbumin-specific IgG2a. PE*-CpG-dG6 increased ovalbumin-specific IgG2a but also reduced BAL fluid eosinophils and airway hyperresponsiveness. PS-CpG increased ovalbumin-specific IgG2a, reduced airway inflammation and airway hyperresponsiveness. PS-CpG-dG6 was less effective than PS-CpG on airway inflammation and airway hyperresponsiveness. In pre-sensitized mice, PE-CpG required not only 3' dG6-run but also the modification of two PS linkages at 5' terminus to inhibit features of asthma. PS-CpG was strong enough to inhibit asthma but PS-CpG-dG6 was less effective.
Animals ; Anti-Asthmatic Agents/*therapeutic use ; Asthma/*drug therapy/physiopathology ; Bronchial Hyperreactivity/drug therapy ; Bronchoalveolar Lavage Fluid/immunology ; Deoxyguanosine/*analogs & derivatives/*chemistry ; Female ; Immunoglobulin G/metabolism ; Interleukin-12/analysis ; Interleukin-4/analysis ; Interleukin-5/analysis ; Lung/pathology ; Mice ; Mice, Inbred BALB C ; Oligodeoxyribonucleotides/*therapeutic use ; Phosphorothioate Oligonucleotides/therapeutic use ; Splenomegaly/pathology