85405-01-2

Usage

Uses

Used in Biochemistry Research:
(6S,8S)-3-(2-deoxy-alpha-D-erythro-pentofuranosyl)-8-hydroxy-6-methyl-4,6,7,8-tetrahydropyrimido[1,2-a]purin-10(3H)-one is utilized in the study of glyoxal adduct formation in Deoxyguanosine. This application is significant for understanding the chemical reactions and modifications that occur within biological systems, particularly in the context of DNA and RNA structures. The study of such adducts can provide insights into the mechanisms of mutagenesis, DNA repair, and the potential implications for genetic stability and disease development.

Upstream product

• 123-73-9 crotonaldehyde 
• 59435-85-7 α-acetoxy-N-nitrosopyrrolidine 
• 961-07-9 2'-Deoxyguanosine 
• 151900-41-3 6-(2-(trimethylsilyl)ethoxy)-2-fluoropurine-9-(2'-deoxyriboside) 

Relevant academic research and scientific papers

Identification of paraldol-deoxyguanosine adducts in DNA reacted with crotonaldehyde

Crotonaldehyde (1) is a mutagen and carcinogen, but its reactions with DNA have been only partially characterized. In a previous study, we found that substantial amounts of 2-(2hydroxypropyl)-4-hydroxy-6-methyl-1,3-dioxane (paraldol, 7), the dimer of 3-hydroxybutanal (8), were released upon enzymatic or neutral thermal hydrolysis of DNA that had been allowed to react 0with crotonaldehyde. We have now characterized two paraldol-deoxyguanosine adducts in this DNA: N2-[2-(2-hydroxypropyl)-6-methyl-1,3-dioxan-4-yl]deoxyguanosine (N2-paraldoldG, 13) and N2-[2-(2-hydroxypropyl)-6-methyl-1,3-dioxan-4-yl]deoxyguanylyl-(5'-3')-thymi dine [N2-paraldol-dG-(5'-3')-thymidine, 14]. Four diastereomers of N2-paraldol-dG (13) were observed. Their overall structures were determined by 1H NMR, by MS, and by reaction of paraldol with deoxyguanosine and DNA. 1H NMR data showed that two diastereomers had all equatorial substituents in the dioxane ring, while two others had an axial 6-methyl group. Preparation of paraldol with the (R)- or (S)-configuration at the 6-position of the dioxane ring and the carbinol carbon of the 2-(2-hydroxypropyl) group allowed partial assignment of the absolute configurations of N2-paraldol-dG (13). Four diastereomers of N2-paraldol-dG-(5'-3')thymidine (14) were observed. Their overall structure was determined by 1H NMR, MS, and hydrolysis with snake venom or spleen phosphodiesterase. Reactions of nucleosides and nucleotides with paraldol demonstrated that adducts were formed only from deoxyguanosine and its monophosphates. Experiments with DNA that had been reacted with crotonaldehyde indicated that N2-paraldol-dG-containing adducts in DNA are relatively resistant to enzymatic hydrolysis. The results of this study demonstrate that the reaction of crotonaldehyde with DNA is more complex than previously recognized and that stable N2-paraldol-dG adducts are among those that should be considered in assessing mechanisms of crotonaldehyde mutagenicity and carcinogenicity.

Ultrasensitive simultaneous quantification of 1, N 2-etheno-2'- deoxyguanosine and 1, N 2-propano-2'-deoxyguanosine in DNA by an online liquid chromatography-electrospray tandem mass spectrometry assay

Exocyclic DNA adducts produced by exogenous and endogenous compounds are emerging as potential tools to study a variety of human diseases and air pollution exposure. A highly sensitive method involving online reverse-phase high performance liquid chromatography with electrospray tandem mass spectrometry detection in the multiple reaction monitoring mode and employing stable isotope-labeled internal standards was developed for the simultaneous quantification of 1,N2-etheno-2'-deoxyguanosine (1,N 2-εdGuo) and 1,N2-propano-2'-deoxyguanosine (1,N 2-propanodGuo) in DNA. This methodology permits direct online quantification of 2'-deoxyguanosine and ca. 500 amol of adducts in 100 μg of hydrolyzed DNA in the same analysis. Using the newly developed technique, accurate determinations of 1,N2-etheno-2'-deoxyguanosine and 1,N 2-propano-2'-deoxyguanosine levels in DNA extracts of human cultured cells (4.01 ± 0.32 1,N2-εdGuo/108 dGuo and 3.43 ± 0.33 1,N2-propanodGuo/108 dGuo) and rat tissue (liver, 2.47 ± 0.61 1,N2-εdGuo/108 dGuo and 4.61 ± 0.69 1,N2-propanodGuo/108 dGuo; brain, 2.96 ± 1.43 1,N2-εdGuo/108 dGuo and 5.66 ± 3.70 1,N2-propanodGuo/108 dGuo; and lung, 0.87 ± 0.34 1,N2-εdGuo/108 dGuo and 2.25 ± 1.72 1,N2-propanodGuo/108 dGuo) were performed. The method described herein can be used to study the biological significance of exocyclic DNA adducts through the quantification of different adducts in humans and experimental animals with pathological conditions and after air pollution exposure.

Reaction of α-Acetoxy-N-nitrosopyrrolidine with Deoxyguanosine and DNA

We investigated the reactions of α-acetoxy-N-nitrosopyrrolidine (α-acetoxyNPYR) with dGuo and DNA. α-AcetoxyNPYR is a stable precursor to the major proximate carcinogen of NPYR, α-hydroxyNPYR (3). Our goal was to develop appropriate conditions for the analysis of DNA adducts of NPYR formed in vivo. Products of the α-acetoxyNPYR-dGuo reactions were analyzed directly by HPLC or after treatment of the reaction mixtures with NaBH3CN. Products of the α-acetoxyNPYR-DNA reactions were released by enzymatic or neutral thermal hydrolysis of the DNA, then analyzed by HPLC. Alternatively, the DNA was treated with NaBH3CN prior to hydrolysis and HPLC analysis. The reactions of α-acetoxyNPYR with dGuo and DNA were complex. We have identified 13 products of the dGuo reaction - 6 of these were characterized in this reaction for the first time. They were four diastereomers of N2-(3-hydroxybutylidene)-dGuo (20,21), 7-(N-nitrosopyrrolidin-2-yl)Gua (2), and 2-(2-hydroxypyrrolidin-1-yl)deoxyinosine (12). Adducts 20 and 21 were identified by comparison to standards produced in the reaction of 3-hydroxybutanal with dGuo. Adduct 2 was identified by its spectral properties while adduct 12 was characterized by comparison to an independently synthesized standard. With the exception of adduct 2, all products of the dGuo reactions were also observed in the DNA reactions. The major product in both the dGuo and DNA reactions was N2-(tetrahydrofuran-2-yl)dGuo (10), consistent with previous studies. Several other previously identified adducts were also observed in this study. HPLC analysis of reaction mixtures treated with NaBH3CN provided improved conditions for adduct identification, which should be useful for in vivo studies of DNA adduct formation by NPYR.

Stereospecific synthesis of oligonucleotides containing crotonaldehyde adducts of deoxyguanosine

Crotonaldehyde reacts with DNA to form two diastereomeric 1,N2 cyclic adducts of deoxyguanosine. A synthesis of the two diastereomeric deoxynucleosides has been achieved by reaction of mixed diastereomers of 4-amino-1,2-pentanediol with 2-fluoro-O6-(trimethylsilylethyl)-deoxyinosine. The resulting N2-(1-methyl-3,4-dihydroxybutyl)-deoxyguanosine was treated with NaIO4, cleaving the vicinal diol to the aldehyde. Spontaneous cyclization gave the two diastereomers of the crotonaldehyde-adducted nucleoside that were readily separated by HPLC. The absolute configurations were assigned by an enantiospecific synthesis of one diastereomer from (S)-3-aminobutanoic acid. The synthetic strategy has been extended to preparation of a site-specifically adducted oligonucleotide by reaction of the mixed diastereomers of 4-amino-1,2-pentanediol with an 8-mer oligonucleotide containing 2-fluoro-O6-(trimethylsilylethyl)-deoxyinosine. The diastereomeric oligonucleotides were separated by HPLC and absolute configurations of the adducts were established by enzymatic digestion to the adducted nucleosides.