A facile solid phase synthesis of 2′- and 3′-aminonucleoside triphosphates

Schoetzau, Thomas; Holletz, Torsten; Cech, Dieter

doi:10.1039/cc9960000387

Cited by 15 publications

(11 citation statements)

References 10 publications

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“…Thus, we synthesized 3'-modified chain terminators in the thymidine series starting from 3'-azido-3'deoxythymidine (AZT; 1) (Scheme 1). Usually polymer-bound triphenylphosphine is suitable for the reduction of azido-nucleosides into the corresponding amino-nucleosides [11] [12]. In this case, the generated phosphine imide is hydrolyzed by concentrated aqueous ammonia.…”

mentioning

confidence: 99%

Synthesis of 3′-Sugar- and Base-Modified Nucleotides and Their Application as Potent Chain Terminators in DNA Sequencing

Stolze¹,

Koert²,

Klingel³

et al. 1999

HCA

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Two 3′‐modified and three base‐modified ddNTPs were synthesized and tested with several DNA polymerases for incorporation activity. Starting from 3′‐azido‐3′‐deoxythymidine (AZT; 1), we were able to produce 3′‐deoxy‐3′‐isocyanato‐thymidine and 3′‐deoxy‐3′‐isothiocyanatothymidine (3 and 4, resp.) in a rapid synthesis based on the solid‐support approach (Scheme 1). These 3′‐functionalities could be used to attach a spacer molecule via urea and thiourea groups, respectively. Since the thus‐obtained tethered nucleotides 7 and 8 can be used to label with fluorescent dyes (cf. Scheme 5), they are convenient building blocks for practical applications in DNA sequencing. Furthermore, we synthesized, via 14 (Scheme 2), 17 (Scheme 3), and 19 (Scheme 4), the N4‐modified dideoxycytidine 5′‐triphosphate dye derivatives 22, 23, and 24, respectively, with different lengths of linkers between the base residue and the dye (Scheme 5). Base‐specific termination for the derivatives 22 and 24 was demonstrated (Fig. 2a and 2b).

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confidence: 99%

Synthesis of 3′-Sugar- and Base-Modified Nucleotides and Their Application as Potent Chain Terminators in DNA Sequencing

Stolze¹,

Koert²,

Klingel³

et al. 1999

HCA

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“…30 It has been reported that insoluble polymeric reagents can be used in large excess to complete this conversion into an alkyl azide. [32][33][34] In Polymer-supported triphenylphosphine has previously been used for the Staudinger reaction, where the conversion of an organoazide (R-N 3 ) by a tertiary phosphine produced an iminophosphorane (R 3 P=NR) that hydrolyzed to give primary amines.…”

Section: Inmentioning

confidence: 99%

“…Polymer-supported triphenylphosphine has previously been used for the Staudinger reaction, where the conversion of an organoazide (R-N 3 ) by a tertiary phosphine produced an iminophosphorane (R 3 P=NR) that hydrolyzed to give primary amines. [32][33][34] In Step E, acetyl hydrazine, in a mixture of glacial acetic acid and methanol, was used to form the acetylhydrazides (6a/b), and the final step (F) was the formation of the triazole ring by thermolysis on the GC injector. The solutions of (6a/b) in acetonitrile could be stored at 4°C -8°C but it was found that the benzodiazepine derivatives were not produced after several months of storage.…”

Section: Inmentioning

confidence: 99%

An approach to shortening the timeframe between the emergence of new compounds on the drugs market and the availability of reference standards: The microscale syntheses of nitrazolam and clonazolam for use as reference materials, utilizing polymer‐supported reagents

Dowling

Kavanagh

Eckhardt³

et al. 2018

Drug Testing and Analysis

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Nitrazolam and clonazolam are 2 designer benzodiazepines available from Internet retailers. There is growing evidence suggesting that such compounds have the potential to cause severe adverse events. Information about tolerability in humans is scarce but typically, low doses can be difficult to administer for users when handling bulk material. Variability of the active ingredient in tablet formulations can also be of a concern. Customs, toxicology and forensic laboratories are increasingly encountering designer benzodiazepines, both in tablet and powdered form. The unavailability of reference standards can impact on the ability to identify these compounds. Therefore, the need arises for exploring in-house approaches to the preparation of new psychoactive substances (NPS) that can be carried out in a timely manner. The present study was triggered when samples of clonazolam were received in powdered and tablet form at a time when reference material for this drug was commercially unavailable. Therefore, microscale syntheses of clonazolam and its deschloro analog nitrazolam were developed utilizing polymer-supported reagents starting from 2-amino-2'-chloro-5-nitrobenzophenone (clonazolam) and 2-amino-5-nitrobenzophenone (nitrazolam). The final reaction step forming the 1,2,4-triazole ring moiety was performed within a gas chromatography-mass spectrometry (GC-MS) injector. A comparison with a preparative scale synthesis of both benzodiazepine derivatives showed that microscale synthesis might be an attractive option for a forensic laboratory in terms of time and cost savings when compared with traditional methods of synthesis and when qualitative identifications are needed to direct forensic casework. The reaction by-product profiles for both the micro and the preparative scale syntheses are also presented.

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“…[2][3][4][5][6][7][8][9] Among these, a liquid-phase synthesis of 5′-TPs by reaction of the oligonucleotide with 2-chloro-4H-1,3,2-benzodioxaphosphorin-4-one (salicyl phosphorochloridite) developed by Ludwig and Eckstein 10) is well known.…”

mentioning

confidence: 99%

“…4,9,11) A solid-phase adaptation of the salicyl phosphorochloridite method of Ludwig and Eckstein 10) has been developed by Schmidt et al,12) who used it to synthesize the 5′-TP of a 25-mer RNA.…”

mentioning

confidence: 99%

Improved Method for the Solid-Phase Synthesis of Oligoribonucleotide 5′-Triphosphates

Nagata

Takagaki

Wada

2012

CHEMICAL & PHARMACEUTICAL BULLETIN

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We have developed an improved solid-phase method for the synthesis of 5′-triphosphates (5′-TPs) of oligoribonucleotides. The method is based on the use of salicyl phosphorochloridite as the phosphitylating reagent and the improvement is characterized by the use of the highly reactive pyrophosphorylating reagent tris(tetra-n-butylammonium) hydrogen pyrophosphate instead of the conventional tri-n-butylammonium salt for the nucleophilic substitution reaction to form the cyclic ester intermediate. The improved method can be used to generate oligoribonucleotide 5′-TPs efficiently and reproducibly.Key words 5′-triphosphate; oligoribonucleotide; solid phase; RNA; triphosphorylation 5′-Triphosphates (5′-TPs) of nucleosides and nucleic acids play an important role in biochemical reactions and have many research and medical applications. Thus, nucleoside 5′-TPs have long been known to be substrates for enzymatic DNA and RNA synthesis. A more recent discovery is the fact that the immune response triggered by the binding of 5′-TPs of RNA oligomers to retinoic acid-induced protein I (RIG-I) works together in synergy with gene silencing mediated by small interfering RNAs to promote apoptosis in tumor cells. 1)As for applications, 5′-TPs of DNA and RNA oligomers are used as substrates for ligation in vitro, while modified nucleoside triphosphates are used as antiviral medicines and diagnostic reagents. The demand for 5′-TPs of DNA and RNA is increasing and will most likely be met by chemical synthetic rather than enzymatic methods because of advantages resulting from the ability to introduce chemically modified nucleosides at any point in the sequence as well as the practicality of scaling up the synthesis. Chemical methods for the synthesis of 5′-TPs of RNA in particular have the additional advantage that they allow the production of highly pure RNA.A number of chemical syntheses of 5′-TPs of DNA and RNA have been reported. 2-9) Among these, a liquid-phase synthesis of 5′-TPs by reaction of the oligonucleotide with 2-chloro-4H-1,3,2-benzodioxaphosphorin-4-one (salicyl phosphorochloridite) developed by Ludwig and Eckstein 10) is well known.11) However, there are few reports of the solid-phase synthesis of DNA and RNA 5′-TPs. 4,9,11) A solid-phase adaptation of the salicyl phosphorochloridite method of Ludwig and Eckstein 10) has been developed by Schmidt et al.,12) who used it to synthesize the 5′-TP of a 25-mer RNA.For triphosphorylation experiments in the present study, we synthesized a 25-mer RNA oligomer by the 2-cyanoethoxymethyl (CEM) method of RNA synthesis developed in our laboratory. [13][14][15] In this method, the CEM group is used as the 2′-hydroxyl protecting group. Because the CEM group is small, it offers minimal steric hindrance and so allows an increased yield at each coupling step. As a result, and also because of the ease of removal of the CEM group under mild conditions after synthesis, RNA can be synthesized more simply and easily by the CEM method than by methods involving conventional 2′-hydroxyl protect...

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A facile solid phase synthesis of 2′- and 3′-aminonucleoside triphosphates

Abstract: A convenient and time-saving solid phase synthesis of different 2'and 3'-amino-functionalized nucleoside-5'-triphosphates using polymer bounded

Cited by 15 publications

References 10 publications

Synthesis of 3′-Sugar- and Base-Modified Nucleotides and Their Application as Potent Chain Terminators in DNA Sequencing

Synthesis of 3′-Sugar- and Base-Modified Nucleotides and Their Application as Potent Chain Terminators in DNA Sequencing

An approach to shortening the timeframe between the emergence of new compounds on the drugs market and the availability of reference standards: The microscale syntheses of nitrazolam and clonazolam for use as reference materials, utilizing polymer‐supported reagents

Improved Method for the Solid-Phase Synthesis of Oligoribonucleotide 5′-Triphosphates

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