Oligonucleotides labeled with stable isotopes and a method for detecting the same

Abstract

Antisense oligonucleotide sequences which enable the measurement of the distribution and structure of antisense oligonucleotide drugs in the body, with lapse of time, and a method of detecting these sequences are provided. The antisense chains have a natural or non-natural nucleotide or peptide nucleic acid as a structural unit in which carbon atoms and nitrogen atoms are substituted by 13 C and 15 N, respectively, and the antisense chains can be detected by nuclear magnetic resonance spectroscopy (NMR) such as 15 N— 1 H or 13 C— 1 H hereto nuclear multiple quantum coherence spectroscopy.

1 - 10 . (canceled)  
     
     
         11 . An antisense drug using an oligonucleotide sequence or peptide nucleic acid sequence comprising a single strand RNA or single strand DNA containing at least one nucleotide or peptide nucleic acid as a structural unit, in which at least one structural C atom is substituted by  13 C, and at least one structural N atom is substituted by  15 N, having 10 to 100 bases, said oligonucleotide being 
 i) a phosphodiester oligonucleotide wherein 3′-OH and 5′-OH in ribose or deoxyribose are cross-linked by phosphodiester bonds,    ii) a phosphorothioate oligonucleotide wherein one or two non cross-linked oxygen atoms in the phosphodiester bonds in said phosphodiester oligonucleotide are substituted by sulfur atoms, or    iii) a methylphosphonate oligonucleotide wherein oxygen atoms in the hydroxyl groups in the phosphodiester bonds in said phosphodiester oligonucleotide are substituted by methyl groups, and    said peptide nucleic acid having bases, i.e., purine or pyrimidine, and    said bases being linked together by peptide bonds to form a 2-aminoethylglycine backbone,    wherein more than 90 % of the carbon atoms are substituted by  13 C and more than 90 % of the nitrogen atoms are substituted by  15 N in every structural unit, i.e., nucleotide or peptide nucleic acid.    
     
     
         12 . An antisense drug using an oligonucleotide sequence or peptide nucleic acid sequence according to  claim 11 , wherein all carbon atoms are substituted by  13 C and all nitrogen atoms are substituted by  15 N in every structural unit, i.e., nucleotide or peptide nucleic acid.  
     
     
         13 . A method of detecting an antisense drug using an oligonucleotide sequence containing stable isotopes or a peptide nucleic acid sequence containing stable isotopes, or decomposition products thereof, comprising a step of subjecting a subject animal, to which the antisense drug using the oligonucleotide sequence or peptide nucleic acid sequence according to  claim 11  is administered, to magnetic resonance imaging.  
     
     
         14 . A method of detecting an antisense drug using an oligonucleotide sequence containing stable isotopes or a peptide nucleic acid sequence containing stable isotopes, or decomposition products thereof, comprising a step of subjecting a subject animal, to which the antisense drug using the oligonucleotide sequence or peptide nucleic acid sequence according to  claim 12  is administered, to magnetic resonance imaging.