Date of Award

7-1-1989

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Department

Chemistry

Abstract

The use of modified analogs of 2$\sp\prime$-deoxyuridine as probes that incorporate into oligonucleotides are discussed. These probes can be useful in many areas including diagnostics, sequencing and therapeutics, and will be discussed later in more detail.The development of DNA probes is important in the diagnostic area because the use of the probes can diminish dramatically the time needed to diagnose an infection (from weeks to hours). Use of DNA probes for sequencing the genomes of hereditary diseases can help find cures. Fighting infectious disease by blocking replication of the virus using DNA probes is a therapeutic use. These are only some uses of DNA probe technology.The modifications of 2$\sp\prime$-deoxyuridine in this investigation are all on C-5. Originally, the focus was on building side chains that were linked to C-5 through an olefin linkage. As an example, N-methylacrylamide was coupled to mercurated 2$\sp\prime$-deoxyuridine.A considerable amount of effort was spent trying to transform a relatively unreactive terminal olefin, 5-(2-propen-1-yl)-2$\sp\prime$-deoxyuridine, into a more reactive species. Thiolacetic acid was coupled to 5-(2-propen-1-yl)-2$\sp\prime$-deoxyuridine under a number of different catalysts to produce a protected mercaptan, 5-(3-oxo-2-thiabutyl)-2$\sp\prime$-deoxyuridine. This transformation was not successful under visible or ultraviolet light, benzoyl peroxide, azobis(isobutyrylnitrile) (AIBN), or (W(C$\sb5$H$\sb5$)(CO)$\sb3$) $\sb2$.The synthesis and purification of 5-hydroxymethyl-2$\sp\prime$-deoxyuridine was also undertaken. 5-Hydroxymethyl-2$\sp\prime$-deoxyuridine was not purified successfully. Reactions using 5-hydroxymethyl-2$\sp\prime$-deoxyuridine were performed with the reagent laced with 2$\sp\prime$-deoxyuridine and fluoride contaminants. 5-Hydroxymethyl-2$\sp\prime$-deoxyuridine was reacted with thiolacetic acid using various catalytic conditions. Unfortunately, the reactions yielded either too little product to analyze or too many products that were unresolvable.A new reaction was discovered and is an integral part of this dissertation. It involves the coupling of a disulfide to 5-chloromercuri-2$\sp\prime$-deoxyuridine. It is mediated by Pd (II) as is the olefin coupling. The reaction is complete in a few hours and the product is obtained in high yield. The reaction is applicable to a wide range of disulfides. We have successfully linked butyl, phenyl, methyl propionate disulfides as well as trifluoroacetylcystamine to 2$\sp\prime$-deoxyuridine. More work is needed to fully understand the chemistry of this reaction.

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