Antiviral azide-containing compounds
An azide and carbohydrate technology, applied in the field of pharmaceutical compositions of salts, can solve problems such as toxic side effects, bone marrow suppression, abnormal liver function and the like
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Embodiment 1
[0187] Example 1: Labeling of HIV with azide-modified biomolecules
[0188] HIV in a T-150 flask NL4-3 Transfect CEMx174 cells and monitor virus production by reverse transcriptase activity until peak virus production is reached (typically 7-9 days after transfection). Prior to transfection, CEMx174 cells were spiked with the following azide-modified biomolecules: 15-azidopentadecanoic acid (50-100 μM), 12-azidododecanoic acid (50-100 μM ), tetraacetylated N-azidoacetylgalactosamine (20-40 μM) and tetraacetylated N-azidoacetyl-D-mannosamine (20-40 μM).
[0189] Infected cells were harvested at 12, 24, 72 hours and 14 days. Harvested cells are detached and lysed. Cell lysates were then mixed with Detection reagent, tetramethylrhodamine (TAMRA) alkyne and Protein Reaction Buffer Kit (Invitrogen, Carlsbad, CA) was mixed together. Cell lysate samples were run on one-dimensional gels to monitor changes in azide-labeled proteins over time ( figure 1 ).
[0190] Labeled vir...
Embodiment 2
[0192] Example 2: Inhibition of HIV infectivity
[0193] To examine the effect of the azide-modified biomolecules on the innate biology of the virus, azide-tagged viruses were isolated from transfected cells and tested in cell infection studies. Unlabeled HIV (concentration 1 / 100 of the test sample) was used as a control. Viral load was normalized to p24 abundance and viruses were incubated for 12 hours on a reporter cell line (TZM / BI). TZM / BI is a genetically engineered HeLa cell line expressing CD4, CXCR4 and CCR5 and containing Tat-inducible luciferase and β-Gal reporter genes. Virus infectivity was determined by measuring cellular luciferase activity with 2 different luciferase reagents. Results using a single-cycle replication system showed that biomolecules modified with the azides (specifically, 12-azidododecanoic acid, and to a lesser extent, 15-azidopentadecanoic acid and tetraacetylated N-azidoacetylgalactosamine)-tagged viruses had profound effects on viral infec...
Embodiment 3
[0194] Example 3: Toxicity Profile
[0195] Little or no toxicity was observed in different eukaryotic cell lines with these azide-modified biomolecules, suggesting that these compounds have a minimal toxicity profile,
[0196] and support their use in therapeutic settings.
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