ABSTRACT
Aminoalkyl and alkyne-tagged PNA HyBeacons have been synthesized, labeled with fluorescein via conventional amide bond or triazole formation (click chemistry) and used to detect single nucleotide polymorphisms (SNPs) implicated in statin-induced myopathy. The PNA HyBeacons gave much better mismatch/mutant discrimination than conventional DNA HyBeacons but smaller fluorescence changes on melting.
Subject(s)
DNA Mutational Analysis/methods , Hydroxymethylglutaryl-CoA Reductase Inhibitors/adverse effects , Muscular Diseases/chemically induced , Muscular Diseases/genetics , Oligonucleotide Probes/chemistry , Peptide Nucleic Acids/chemistry , Click Chemistry , Fluorescent Dyes , Genotyping Techniques/methods , Humans , Mass Spectrometry , Oligonucleotide Probes/chemical synthesis , Polymorphism, Single Nucleotide , Transition TemperatureABSTRACT
Rapid controlled self-assembly makes DNA ideal for building nanostructures. A problem using hybridized intermediates in hierarchic assembly is their thermodynamic lability. We demonstrate a click-fixation technology by which robust hexagonal DNA modules can be made. This principle is applicable to a wide variety of DNA nanoconstructs.
Subject(s)
DNA/chemistry , Nanostructures/chemistry , Electrophoresis, Polyacrylamide Gel , Fluorescence Resonance Energy Transfer , Nucleic Acid Hybridization , Oligonucleotides/chemistryABSTRACT
Structurally diverse, sugar-modified, thymine-containing nucleoside phosphonic acids were evaluated for their ability to inhibit thymidine phosphorylase (TP, EC 2.4.2.4) purified from spontaneous T-cell lymphomas of an inbred Sprague-Dawley rat strain. From a large set of tested compounds, among them a number of pyrrolidine-based derivatives, 10 nucleotide analogues with IC(50) values below 1 microM were selected. Out of them, four compounds strongly inhibited the enzyme with IC(50) values lying in a range of 11-45 nM. These most potent compounds might be bi-substrate analogues.
Subject(s)
Lymphoma, T-Cell/enzymology , Nucleosides/chemistry , Organophosphonates/chemistry , Thymidine Phosphorylase/antagonists & inhibitors , Animals , Dose-Response Relationship, Drug , Humans , Inhibitory Concentration 50 , Nucleosides/pharmacology , Organophosphonates/pharmacology , Rats , Rats, Sprague-Dawley , Structure-Activity Relationship , Thymidine Phosphorylase/metabolismABSTRACT
Two series of thymidine derivatives with additional nucleobases/aromatics attached to either the 5'(S)-C- or the 5-position were prepared by epoxide opening and/or "click chemistry" cycloaddition protocols and introduced into DNA duplexes. Interstrand base-base communication in the minor groove and intrastrand stacking interactions in the major groove were detected.
Subject(s)
DNA/chemistry , Thymidine/analogs & derivatives , Nucleic Acid Conformation , Nucleic Acid Denaturation , Pyrimidine Nucleosides/chemical synthesis , Pyrimidine Nucleosides/chemistry , Temperature , Triazoles/chemistryABSTRACT
A synthetic strategy towards new aromatic nucleoside derivatives introducing additional aromatic functionality placed in the major groove of a modified DNA duplex is presented. The functionalities are introduced using Click Chemistry conditions and found to increase the overall duplex stability.
Subject(s)
DNA/chemistry , Pyrimidine Nucleosides/chemical synthesis , Biochemistry/methods , Models, Molecular , Nucleic Acid Conformation , Pyrimidine Nucleosides/chemistryABSTRACT
Among the structurally diverse nucleoside phosphonic acids, several compounds possessing strong antiviral properties have been found. Our effort in this area was focused to the synthesis of novel compounds - pyrrolidine-based nucleoside phosphonic acids and their derivatives.
Subject(s)
Antiviral Agents/chemical synthesis , Nucleosides/chemical synthesis , Nucleotides/chemical synthesis , Pyrrolidines/chemistry , Antiviral Agents/chemistry , Nucleosides/chemistry , Nucleotides/chemistry , Organophosphonates/chemistryABSTRACT
A number of structurally diverse nucleoside phosphonic acids have been tested against human recombinant thymidine phosphorylase and human platelets supernatant using 2'-deoxy-5-nitrouridine as the substrate. We have selected several inhibitors working at micromolar level as lead structures for further evaluation.
Subject(s)
Enzyme Inhibitors/chemistry , Nucleosides/chemistry , Nucleosides/pharmacology , Organophosphonates/chemistry , Thymidine Phosphorylase/antagonists & inhibitors , Animals , Blood Platelets/enzymology , CHO Cells , Cricetinae , Cricetulus , Enzyme Inhibitors/pharmacology , Humans , Structure-Activity Relationship , Thymidine Phosphorylase/chemistryABSTRACT
Click chemistry has been used to covalently cross-link complementary DNA strands between bases to form very stable duplexes. Several alkyne- and azide-modified uracil monomers were used to evaluate the effect of the linkers on the efficiency of the click reaction. All cross-linked duplexes had much higher thermal stabilities than non-cross-linked ones, with increases in melting temperature of up to 30 degrees C. In some cases, the conversion was near-quantitative, and the reaction was complete in 5 min.
Subject(s)
Cross-Linking Reagents/chemistry , DNA/chemistry , Alkynes/chemistry , Azides/chemistry , Circular Dichroism , Deoxyuridine/chemistry , Electrophoresis, Capillary , Molecular Structure , Oligonucleotides/chemical synthesis , Oligonucleotides/chemistry , Time FactorsABSTRACT
A general protocol for converting alkyl and aryl halides into azides and for converting these in situ into 1,4-disubstituted triazoles was applied with 5-ethynyl-2'-deoxyuridine. This afforded three modified 2'-deoxyuridine analogues with either unsubstituted or 1-phenyl-/1-benzyl-substituted triazoles in their 5-positions. Modelling demonstrates coplanarity of the two heteroaromatic rings, and UV spectroscopy showed the uracil pK(a) values to be almost unchanged. The three nucleosides were introduced into nonamer oligonucleotides by phosphoramidite chemistry. The heteroaromatic triazoles became positioned in the major grooves of the short dsDNA and DNA-RNA duplexes. While single modifications led to decreased duplex stability, the stacking of four consecutive modifications led to enhanced duplex stability, especially for DNA-RNA duplexes. The duplex structures were studied by CD spectroscopy and molecular dynamics simulations, which supported the conjecture that the duplex stabilizing effect is due to efficient stacking of the heteroaromatic triazoles.
Subject(s)
Deoxyuridine/chemical synthesis , Nucleic Acids/chemistry , Triazoles/chemistry , Circular Dichroism , Deoxyuridine/chemistry , Models, Molecular , Molecular Structure , SpectrophotometryABSTRACT
The present work relates to the synthesis of pyrrolidine nucleoside analogs. Starting from malic acid, we have elaborated a high-yield synthesis of racemic and enantiomeric N-protected 3-pyrrolidinols and their O-mesyl derivatives as key compounds for alkylations of purine and pyrimidine nucleobases. On varying base and solvent, we have found conditions providing both satisfactory N-/O-regioisomeric ratio and acceptable yield for pyrimidine compounds.