Synthesis of 2′‐Aminoalkyl‐Substituted Fluorinated Nucleobases and Their Influence on the Kinetic Properties of Hammerhead Ribozymes
Identifieur interne : 000B76 ( Istex/Checkpoint ); précédent : 000B75; suivant : 000B77Synthesis of 2′‐Aminoalkyl‐Substituted Fluorinated Nucleobases and Their Influence on the Kinetic Properties of Hammerhead Ribozymes
Auteurs : Astrid E. Klöpffer ; Joachim W. Engels [Allemagne]Source :
- ChemBioChem [ 1439-4227 ] ; 2004-05-03.
English descriptors
- Teeft :
- Ach2, Amersham biosciences, Analogue, Anhydrous, Anhydrous pyridine, Aqueous solution, Artificial nucleosides, Base pair, Bch2, Benzimidazole, Black bars, Carom, Catalytic activity, Cdcl3, Chem, Chembiochem, Cleavage, Cleavage rates, Cleavage reactions, Crude product, Dropwise, Dropwise addition, Duplex, Engels, Equiv, Gene expression, Gmbh, Hammerhead, Hammerhead ribozyme, Hammerhead ribozymes, Harom, Hphth, Hydrogen bonds, Kgaa, Kinetic analysis, Kinetic parameters, Kinetic properties, Methyl bromoacetate, Mgso4, Mismatch, Mismatch complexes, Mismatch variants, Mmol, Mutation, Natural nucleosides, Nucleobase, Nucleobase analogues, Nucleobases, Nucleoside, Nucleoside analogue, Nucleoside analogues, Organic extracts, Phosphoroamidite, Point mutations, Possible substrates, Primary alcohol, Pyridine, Reaction mixture, Reaction rates, Ribozyme, Ribozymes, Rna, Room temperature, Sichch3, Sodium hydride, Sugar moiety, Tetrabutylammonium fluoride, Thermodynamic data, Universal nucleobases, Unmodified, Unmodified ribozyme, Verlag, Verlag gmbh, Weinheim, Weinheim chembiochem, White foam.
Abstract
Hammerhead ribozymes are ribonucleic acids that catalyse the hydrolytic cleavage of RNA. They interfere with gene expression in a highly specific manner and recognize the mRNA target through Watson–Crick base pairing. To overcome the problem of point mutations (Watson–Crick “mismatches”) occurring in viral genomes, we developed 2′‐aminoethyl‐substituted fluorinated nucleosides, which are universal nucleobases. The highly efficient synthetic pathway, which features a direct phthaloylamination of a primary alcohol under Mitsunobu conditions, leads to modified phosphoroamidites. The 1′‐deoxy‐1′‐(4,6‐difluoro‐1H‐benzimidazol‐1‐yl)‐2′‐(β‐aminoethyl)‐β‐D‐ribofuranose nucleoside analogue does not differentiate between the four natural nucleosides and leads to a RNA duplex that is as stable as the unmodified parent duplex. Upon incorporation into a ribozyme, the analogue's catalytic activity is equal for all four possible substrates, and the cleavage rates for the modified ribozymes are significantly higher (up to a factor of 13) than for the natural Watson–Crick “mismatch” base pairs. In agreement with the thermodynamic data obtained by measurement of the Tm values of the RNA 12‐mers, the cleavage rates for the 2′‐substituted fluorinated benzimidazole derivative 4 are slightly higher than for the corresponding fluorinated benzene derivative 3.
Fluorinated nucleoside analogues and modified RNAs: The optimised synthesis of 2′‐aminoethyl‐substituted fluorinated benzene and benzimidazole nucleosides (see scheme) and their incorporation into oligoribonucleotides is reported. The influence of these universal bases on the thermodynamic stability of RNA 12‐mer duplexes was investigated by UV/Vis melting studies. The effect of these artificial nucleobases on the reaction rate of ribozyme‐catalysed RNA cleavage was studied by kinetic analyses to evaluate their tolerance against point mutations in biological systems. Owing to their improved properties for RNA recognition and their easy incorporation into hammerhead ribozymes, the nucleosides are applicable to a variety of different mRNA targets.
Url:
DOI: 10.1002/cbic.200300809
Affiliations:
Links toward previous steps (curation, corpus...)
Links to Exploration step
ISTEX:3E9E819D7F9B665CF64963FA2876A5D9D78EA7E6Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Synthesis of 2′‐Aminoalkyl‐Substituted Fluorinated Nucleobases and Their Influence on the Kinetic Properties of Hammerhead Ribozymes</title>
<author><name sortKey="Klopffer, Astrid E" sort="Klopffer, Astrid E" uniqKey="Klopffer A" first="Astrid E." last="Klöpffer">Astrid E. Klöpffer</name>
</author>
<author><name sortKey="Engels, Joachim W" sort="Engels, Joachim W" uniqKey="Engels J" first="Joachim W." last="Engels">Joachim W. Engels</name>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">ISTEX</idno>
<idno type="RBID">ISTEX:3E9E819D7F9B665CF64963FA2876A5D9D78EA7E6</idno>
<date when="2004" year="2004">2004</date>
<idno type="doi">10.1002/cbic.200300809</idno>
<idno type="url">https://api.istex.fr/ark:/67375/WNG-XKWPVQLC-9/fulltext.pdf</idno>
<idno type="wicri:Area/Istex/Corpus">000362</idno>
<idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000362</idno>
<idno type="wicri:Area/Istex/Curation">000362</idno>
<idno type="wicri:Area/Istex/Checkpoint">000B76</idno>
<idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">000B76</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title level="a" type="main">Synthesis of 2′‐Aminoalkyl‐Substituted Fluorinated Nucleobases and Their Influence on the Kinetic Properties of Hammerhead Ribozymes</title>
<author><name sortKey="Klopffer, Astrid E" sort="Klopffer, Astrid E" uniqKey="Klopffer A" first="Astrid E." last="Klöpffer">Astrid E. Klöpffer</name>
<affiliation><wicri:noCountry code="subField">(+49) 69‐798‐29148</wicri:noCountry>
</affiliation>
</author>
<author><name sortKey="Engels, Joachim W" sort="Engels, Joachim W" uniqKey="Engels J" first="Joachim W." last="Engels">Joachim W. Engels</name>
<affiliation></affiliation>
<affiliation wicri:level="1"><country wicri:rule="url">Allemagne</country>
</affiliation>
</author>
</analytic>
<monogr></monogr>
<series><title level="j" type="main">ChemBioChem</title>
<title level="j" type="sub">Fluorine in the Life Sciences</title>
<title level="j" type="alt">CHEMBIOCHEM</title>
<idno type="ISSN">1439-4227</idno>
<idno type="eISSN">1439-7633</idno>
<imprint><biblScope unit="vol">5</biblScope>
<biblScope unit="issue">5</biblScope>
<biblScope unit="page" from="707">707</biblScope>
<biblScope unit="page" to="716">716</biblScope>
<biblScope unit="page-count">10</biblScope>
<publisher>WILEY‐VCH Verlag</publisher>
<pubPlace>Weinheim</pubPlace>
<date type="published" when="2004-05-03">2004-05-03</date>
</imprint>
<idno type="ISSN">1439-4227</idno>
</series>
</biblStruct>
</sourceDesc>
<seriesStmt><idno type="ISSN">1439-4227</idno>
</seriesStmt>
</fileDesc>
<profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Ach2</term>
<term>Amersham biosciences</term>
<term>Analogue</term>
<term>Anhydrous</term>
<term>Anhydrous pyridine</term>
<term>Aqueous solution</term>
<term>Artificial nucleosides</term>
<term>Base pair</term>
<term>Bch2</term>
<term>Benzimidazole</term>
<term>Black bars</term>
<term>Carom</term>
<term>Catalytic activity</term>
<term>Cdcl3</term>
<term>Chem</term>
<term>Chembiochem</term>
<term>Cleavage</term>
<term>Cleavage rates</term>
<term>Cleavage reactions</term>
<term>Crude product</term>
<term>Dropwise</term>
<term>Dropwise addition</term>
<term>Duplex</term>
<term>Engels</term>
<term>Equiv</term>
<term>Gene expression</term>
<term>Gmbh</term>
<term>Hammerhead</term>
<term>Hammerhead ribozyme</term>
<term>Hammerhead ribozymes</term>
<term>Harom</term>
<term>Hphth</term>
<term>Hydrogen bonds</term>
<term>Kgaa</term>
<term>Kinetic analysis</term>
<term>Kinetic parameters</term>
<term>Kinetic properties</term>
<term>Methyl bromoacetate</term>
<term>Mgso4</term>
<term>Mismatch</term>
<term>Mismatch complexes</term>
<term>Mismatch variants</term>
<term>Mmol</term>
<term>Mutation</term>
<term>Natural nucleosides</term>
<term>Nucleobase</term>
<term>Nucleobase analogues</term>
<term>Nucleobases</term>
<term>Nucleoside</term>
<term>Nucleoside analogue</term>
<term>Nucleoside analogues</term>
<term>Organic extracts</term>
<term>Phosphoroamidite</term>
<term>Point mutations</term>
<term>Possible substrates</term>
<term>Primary alcohol</term>
<term>Pyridine</term>
<term>Reaction mixture</term>
<term>Reaction rates</term>
<term>Ribozyme</term>
<term>Ribozymes</term>
<term>Rna</term>
<term>Room temperature</term>
<term>Sichch3</term>
<term>Sodium hydride</term>
<term>Sugar moiety</term>
<term>Tetrabutylammonium fluoride</term>
<term>Thermodynamic data</term>
<term>Universal nucleobases</term>
<term>Unmodified</term>
<term>Unmodified ribozyme</term>
<term>Verlag</term>
<term>Verlag gmbh</term>
<term>Weinheim</term>
<term>Weinheim chembiochem</term>
<term>White foam</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">Hammerhead ribozymes are ribonucleic acids that catalyse the hydrolytic cleavage of RNA. They interfere with gene expression in a highly specific manner and recognize the mRNA target through Watson–Crick base pairing. To overcome the problem of point mutations (Watson–Crick “mismatches”) occurring in viral genomes, we developed 2′‐aminoethyl‐substituted fluorinated nucleosides, which are universal nucleobases. The highly efficient synthetic pathway, which features a direct phthaloylamination of a primary alcohol under Mitsunobu conditions, leads to modified phosphoroamidites. The 1′‐deoxy‐1′‐(4,6‐difluoro‐1H‐benzimidazol‐1‐yl)‐2′‐(β‐aminoethyl)‐β‐D‐ribofuranose nucleoside analogue does not differentiate between the four natural nucleosides and leads to a RNA duplex that is as stable as the unmodified parent duplex. Upon incorporation into a ribozyme, the analogue's catalytic activity is equal for all four possible substrates, and the cleavage rates for the modified ribozymes are significantly higher (up to a factor of 13) than for the natural Watson–Crick “mismatch” base pairs. In agreement with the thermodynamic data obtained by measurement of the Tm values of the RNA 12‐mers, the cleavage rates for the 2′‐substituted fluorinated benzimidazole derivative 4 are slightly higher than for the corresponding fluorinated benzene derivative 3.</div>
<div type="abstract" xml:lang="en">Fluorinated nucleoside analogues and modified RNAs: The optimised synthesis of 2′‐aminoethyl‐substituted fluorinated benzene and benzimidazole nucleosides (see scheme) and their incorporation into oligoribonucleotides is reported. The influence of these universal bases on the thermodynamic stability of RNA 12‐mer duplexes was investigated by UV/Vis melting studies. The effect of these artificial nucleobases on the reaction rate of ribozyme‐catalysed RNA cleavage was studied by kinetic analyses to evaluate their tolerance against point mutations in biological systems. Owing to their improved properties for RNA recognition and their easy incorporation into hammerhead ribozymes, the nucleosides are applicable to a variety of different mRNA targets.</div>
</front>
</TEI>
<affiliations><list><country><li>Allemagne</li>
</country>
</list>
<tree><noCountry><name sortKey="Klopffer, Astrid E" sort="Klopffer, Astrid E" uniqKey="Klopffer A" first="Astrid E." last="Klöpffer">Astrid E. Klöpffer</name>
</noCountry>
<country name="Allemagne"><noRegion><name sortKey="Engels, Joachim W" sort="Engels, Joachim W" uniqKey="Engels J" first="Joachim W." last="Engels">Joachim W. Engels</name>
</noRegion>
</country>
</tree>
</affiliations>
</record>
Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Istex/Checkpoint
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000B76 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Istex/Checkpoint/biblio.hfd -nk 000B76 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien |wiki= Sante |area= MersV1 |flux= Istex |étape= Checkpoint |type= RBID |clé= ISTEX:3E9E819D7F9B665CF64963FA2876A5D9D78EA7E6 |texte= Synthesis of 2′‐Aminoalkyl‐Substituted Fluorinated Nucleobases and Their Influence on the Kinetic Properties of Hammerhead Ribozymes }}
![]() | This area was generated with Dilib version V0.6.33. | ![]() |