Preparation of a deoxynucleoside thiophosphoramidite intermediate in the synthesis of nucleoside phosphorodithioates

Farschtschi, Nasser; Gorenstein, David G.

doi:10.1016/s0040-4039(00)88455-5

Cited by 42 publications

(17 citation statements)

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“…With the demonstration that oligonucleotides and various analogues of oligonucleotides exhibit antiviral and anti-oncogene activity, considerable interest has developed around the synthesis, structural characterization and biological activity of various "antisense" oligonucleotide agents (2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20). Oligodeoxyribonucleotides, oligodeoxynucleoside methylphosphonate, phosphorothioate, phosphoroamidate and phosphorodithioate analogues among others have been studied.…”

Section: Introductionmentioning

confidence: 99%

“…Synthesis using phosphoramidite (13)(14)(15) and solid phase chemistry is now well established. Caruthers and coworkers have shown that the dithioate oligodeoxyribonucleotides are nuclease resistant and are 28-600 times more effective in inhibiting HIV reverse transcriptase (ID 50 of 60 nM) than the normal antisense oligonucleotide or the monothio analogue (20).…”

Section: Introductionmentioning

confidence: 99%

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2D¹H and³¹P NMR Spectra and Distorted A-DNA-Like Duplex Structure of a Phosphorodithioate Oligonucleotide

Cho

Zhu

Luxon

et al. 1993

Journal of Biomolecular Structure and Dynamics

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Assignment of the 1H and 31P NMR spectra of a phosphorodithioate modified oligonucleotide decamer duplex, d(CGCTTpS2-AAGCG)2 (10-mer-S; a site of dithioate substitution is designated with the symbols pS2-), was achieved by two-dimensional homonuclear TOCSY, NOESY and 1H-31P Pure Absorption phase Constant time (PAC) heteronuclear correlation spectroscopy. In contrast to the parent palindromic decamer sequence (1) which has been shown to exist entirely in the duplex B-DNA conformation under comparable conditions (100 mM KCl), the dithiophosphate analogue forms a hairpin loop. However, the duplex form of the dithioate oligonucleotide can be stabilized at lower temperatures, higher salt and strand concentration. The solution structure of the decamer duplex was calculated by an iterative hybrid relaxation matrix method (MORASS) combined with 2D NOESY-distance restrained molecular dynamics. These backbone modified compounds, potentially attractive antisense oligonucleotide agents, are often assumed to possess similar structure as the parent nucleic acid complex. Importantly, the refined structure of the phosphorodithioate duplex shows a significant deviation from the parent unmodified, phosphoryl duplex. An overall bend and unwinding in the phosphorodithioate duplex is observed. The structural distortion of the phosphorodithioate duplex was confirmed by comparison of helicoidal parameters and groove dimensions. Especially, the helical twists of the phosphorodithioate decamer deviate significantly from the parent phosphoryl decamer. The minor groove width of phosphorodithioate duplex 10-mer-S varies between 8.4 and 13.3 A which is much wider than those of the parent phosphoryl decamer d(CGCTTAAGCG)2 (4.2 approximately 9.4 A). The larger minor groove width of 10-mer-S duplex contributes to the unwinding of the backbone and indicates that the duplex has an overall A-DNA-like conformation in the region surrounding the dithiophosphate modification.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

2D¹H and³¹P NMR Spectra and Distorted A-DNA-Like Duplex Structure of a Phosphorodithioate Oligonucleotide

Cho

Zhu

Luxon

et al. 1993

Journal of Biomolecular Structure and Dynamics

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“…1) as potential inhibitors of HIV-1 activity. This analog (13)(14)(15)(16)(17)(18)(19)(20) (25,26) was a gift of S. Hughes (National Cancer Institute).…”

Section: Introductionmentioning

confidence: 99%

Inhibition of human immunodeficiency virus activity by phosphorodithioate oligodeoxycytidine.

Marshall

Beaton

Stein

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

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Phosphorothioate oligodeoxynucleotides exert a sequence-independent cytoprotective effect against human immunodeficiency virus type 1 (HIV-1). We now report that phosphorodithioate-containing oligodeoxycytidines are very potent inhibitors of HIV-1 reverse transcriptase in vitro, as they exhibit an increasing inhibitory effect with length and number of phosphorodithioate internucleotide linkages. This inhibitory effect can be at least 30-fold greater with phosphorodithioate oligodeoxycytidine than for the corresponding phosphorothioate analog of similar length. In cell culture, phosphorodithioate oligodeoxycytidines are active inhibitors of syncytia formation and effectively inhibit de novo infection of target cells by HIV-1. Moreover, comparative experiments show that a deoxycytidine phosphorodithioate 14-mer is as effective an inhibitor of de novo infection as a phosphorothioate-containing 28-mer. Such potent inhibition by oligomers of relatively short length makes dithioate analogs an additional class of potential therapeutic agents against acquired immunodeficiency syndrome.Over the past several years, oligodeoxynucleotides and their analogs have been reported to inhibit viral replication and the expression of oncogenes in cell culture (1, 2). Although the pioneering work of Zamecnik and Stephenson (3, 4) used normal oligodeoxynucleotides, recent research has focused on derivatives ( Fig. 1) that have certain desirable features, including nuclease resistance and rapid transport into cells (5-8). Generally these oligomers appear to function by inhibiting translation and splicing, but other inactivation mechanisms have also been found (9).During studies with oligodeoxynucleotides as inhibitors of human immunodeficiency virus type 1 (HIV-1), Matsukura et al. discovered that phosphorothioate oligodeoxycytidines exhibited potent antiviral activity (10); in these oligomers, one of the nonbridging oxygens at each linkage in oligodeoxycytidine, (dC)", has been replaced with sulfur, and individual oligomers are designated S(dC),. Inhibition of HIV-1 reverse transcriptase (RT) (11) or other steps in its life cycle such as binding, fusion, and entry (5, 10) were suggested as possible sites of action. Subsequent studies corroborated these observations and demonstrated that phosphorothioates are more effective than phosphoramidates and methylphosphonates as sequence-nonspecific inhibitors of HIV-1 replication (12).These developments prompted us to examine oligodeoxycytidine analogs bearing one or more phosphorodithioate linkages (Fig. 1) MATERIALS AND METHODSOligodeoxycytidines. , phosphorodithioate-containing (16, 17, 23), and unmodified oligodeoxycytidines (24) were synthesized and characterized by conventional procedures.Inhibition of Purified EHV-1 RT. Twenty-microliter assay mixtures contained 50 mM Tris (pH 8.3); 10 mM MgCl2; 50 mM KCI; 5 mM dithiothreitol; 1.0 ,uM preannealed primed template, 5'-32P-labeled d(GpApTpTpCpApGpCpTpApGpTpCpCpA) primer and d(CpApApApCpTpGpTpGpApTpApCpGpApTpGpGpApCpTpApGpCpTpGpAp...

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“…Since 2-5A is subject to facile degradation by phosphodiesterases (27) including one with 2',5'-phosphodiester bond specificity (28), additional stabilization of internucleotide linkages could be beneficial for the biological activity of 2-5A. Recently the synthesis of phosphorodithioate oligodeoxyribonucleotides by a solid phase methodology has been described (29)(30)(31)(32). Phosphorodithioate internucleotide linkages are isosteric and isoelectronic with natural phosphodiester linkages and are also nuclease resistant (14).…”

Section: \\ \^\mentioning

confidence: 99%

Synthesis and biological activities of a phosphorodithioate analog of 2′, 5′-oligoadenylate

Beigelman¹,

Matulić‐Adamić²,

Haeberli³

et al. 1995

Nucl Acids Res

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Preparation of a deoxynucleoside thiophosphoramidite intermediate in the synthesis of nucleoside phosphorodithioates

Cited by 42 publications

References 10 publications

2D¹H and³¹P NMR Spectra and Distorted A-DNA-Like Duplex Structure of a Phosphorodithioate Oligonucleotide

2D¹H and³¹P NMR Spectra and Distorted A-DNA-Like Duplex Structure of a Phosphorodithioate Oligonucleotide

Inhibition of human immunodeficiency virus activity by phosphorodithioate oligodeoxycytidine.

Synthesis and biological activities of a phosphorodithioate analog of 2′, 5′-oligoadenylate

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Preparation of a deoxynucleoside thiophosphoramidite intermediate in the synthesis of nucleoside phosphorodithioates

Cited by 42 publications

References 10 publications

2D1H and31P NMR Spectra and Distorted A-DNA-Like Duplex Structure of a Phosphorodithioate Oligonucleotide

2D1H and31P NMR Spectra and Distorted A-DNA-Like Duplex Structure of a Phosphorodithioate Oligonucleotide

Inhibition of human immunodeficiency virus activity by phosphorodithioate oligodeoxycytidine.

Synthesis and biological activities of a phosphorodithioate analog of 2′, 5′-oligoadenylate

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2D¹H and³¹P NMR Spectra and Distorted A-DNA-Like Duplex Structure of a Phosphorodithioate Oligonucleotide

2D¹H and³¹P NMR Spectra and Distorted A-DNA-Like Duplex Structure of a Phosphorodithioate Oligonucleotide