US5698687A
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Modified ribozymls
Assignee: Max-Planck-Gesellschaft zur Förderung der Wissenschaften eV
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Filing: 1995-05-04
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Granted: 1997-12-16
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Expires: 2015-05-04
Abstract
Abstract The present invention refers to an RNA molecule with catalytic activity comprising at least one modified nucleoside, wherein the hydroxy group at the 2'-position of the fibose sugar is replaced by a modifier group, selected from halo, sulfhydryl, azido, amino, monosubstituted amino, and disubstituted andno groups, a process for the preparation of modified KNA molecules and the use of modified KNA molecules as therapeutic agents and biocatalysts.
Inventors
- 👤 Fritz Eckstein
- 👤 Wolfgang Pieken
- 👤 Fritz Benseler
- 👤 David B. Olsen
- 👤 David M. Williams
- 👤 Olaf Heindenreich
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 19:35
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/113; C12N15/1131; C12N15/1132; A; A61; A61P; A61P31/00; A61P31/12; A61P31/14; A61P31/18; A61K; A61K38/00; C12N2310/00; C12N2310/10; C12N2310/12; C12N2310/121; C12N2310/122; C12N2310/30; C12N2310/31; C12N2310/315; C12N2310/32; C12N2310/322
Claims Analysis
Total Claims: 38
Claim 1
Independent Claim
58 words
A method for synthesis of an RNA molecule having catalytic activity comprising: incorporating into an RNA chain at least one modified nucleotide, wherein the hydroxy group at the 2' position of the ribose sugar of said nucleotide is replaced by a modifier group selected from the group consisting of halo, azido, amino, monosubstituted amino and disubstituted amino groups.
Claim 2
Independent Claim
58 words
A method for synthesis of an RNA molecule having catalytic activity comprising: incorporating into an RNA chain at least one modified nucleotide, wherein the hydroxy group at the 2' position of the ribose sugar of said nucleotide is replaced by a modifier group selected from the group consisting of halo, azido, amino, monosubstituted amino and disubstituted amino groups.
Claim 3
Dependent Claim
13 words
The method of claim 1 wherein said modifier group is a halo group.
Claim 4
Dependent Claim
13 words
The method of claim 2 wherein said halo group is a fluoro group.
Claim 5
Dependent Claim
13 words
The method of claim 1 wherein said modifier group is an amino group.
Claim 6
Dependent Claim
14 words
The method of claim 1 wherein said modifier group is a monosubstituted amino group.
Claim 7
Dependent Claim
14 words
The method of claim 1 wherein said modifier group is a disubstituted amino group.
Claim 8
Dependent Claim
48 words
The method of claim 2, wherein said synthesis of said RNA chain is carried out by chemical synthesis from nucleotide precursors on a solid support, further comprising the steps of removing said RNA chain from said solid support, and purifying said RNA chain removed from said solid support.
Claim 9
Dependent Claim
33 words
The method of claim 2, wherein said synthesis of said RNA chain is carded out by chemical synthesis from nucleotide precursors in solution and further comprising the step of purifying said RNA chain.
Claim 10
Dependent Claim
19 words
The method of claim 7 or 8, wherein the respeetive phosphoramidites or H-phosphonates are used as said nucleotide precursors.
Claim 11
Dependent Claim
21 words
The method of claim 2, wherein said method further comprises incorporating into said RNA chain at least one modified internucleotidic linkage.
Claim 12
Dependent Claim
14 words
The method of claim 10, wherein said modified intemucleotidic linkage is a phosphorothioate group.
Claim 13
Dependent Claim
27 words
The method of claim 2, wherein said synthesis of said RNA chain is carded out by transcription from a nucleic acid template by a nucleic acid polymerase.
Claim 14
Dependent Claim
25 words
The method of claim 12, wherein said nucleic acid template is a DNA template and said nucleic acid polymerase is a DNA dependent RNA polymerase.
Claim 15
Dependent Claim
23 words
The method of claim 13, wherein said DNA dependent RNA polymerase is selected from the group, consisting of T7, T3 and SP6 polymerase.
Claim 16
Dependent Claim
15 words
The method of claim 14, wherein said DNA dependent RNA polymerase is said T7 polymerase.
Claim 17
Dependent Claim
22 words
The method of claim 13, wherein said synthesis of said RNA chain is carried out in the presence of Mn 2+ ions.
Claim 18
Dependent Claim
48 words
The method of claim 7, wherein said synthesis of said RNA chain is carried out by chemical synthesis from nucleotide precursors on a solid support, further comprising the steps of removing said RNA chain from said solid support, and purifying said RNA chain removed from said solid support.
Claim 19
Dependent Claim
33 words
The method of claim 4, wherein said synthesis of said RNA chain is carried out by chemical synthesis from nucleotide precursors in solution and further comprising the step of purifying said RNA chain.
Claim 20
Dependent Claim
19 words
The method of claim 17 or 18, wherein the respective phosphoramidites or H-phosphonates are used as said nucleotide precursors.
Claim 21
Dependent Claim
21 words
The method of claim 4, wherein said method further comprises incorporating into said RNA chain at least one modified internucleotidic linkage.
Claim 22
Dependent Claim
14 words
The method of claim 20, wherein said modified internucleotidic linkage is a phosphorothioate group.
Claim 23
Dependent Claim
27 words
The method of claim 12, wherein said synthesis of said RNA chain is carried out by transcription from a nucleic acid template by a nucleic acid polymerase.
Claim 24
Dependent Claim
25 words
The method of claim 22, wherein said nucleic acid template is a DNA template and said nucleic acid polymerase is a DNA dependent RNA polymerase.
Claim 25
Dependent Claim
23 words
The method of claim 23, wherein said DNA dependent RNA polymerase is selected from the group, consisting of T7, T3 and SP6 polymerase.
Claim 26
Dependent Claim
14 words
The method of claim 24, wherein said DNA dependent RNA polymerase said T7 polymerase.
Claim 27
Dependent Claim
22 words
The method of claim 23, wherein said synthesis of said RNA chain is carried out in the presence of Mn 2+ ions.
Claim 28
Dependent Claim
39 words
The method of claim 4, 17, 18, or 19, wherein said amino group is incorporated into said RNA chain in the form of a trifluoroacetyl amido group, and said trifluoroacetyl protecting group is subsequently removed by treatment with ammonia.
Claim 29
Dependent Claim
43 words
The method of claim 5 or 6, wherein said synthesis of said RNA chain is carried out by transcription from a DNA template by a DNA dependent RNA polymerase, and wherein said synthesis is carried out in the presence of Mg 2+ ions.
Claim 30
Dependent Claim
48 words
The method of claim 1, wherein said synthesis of said RNA chain is carried out by chemical synthesis from nucleotide precursors on a solid support, further comprising the steps of removing said RNA chain from said solid support, and purifying said RNA chain removed from said solid support.
Claim 31
Dependent Claim
33 words
The method of claim 1, wherein said iynthesis of said RNA chain is carried out by chemical synthesis from nucleotide precursors in solution and further comprising the step of purifying said RNA chain.
Claim 32
Dependent Claim
19 words
The method of claim 29 or 30, wherein the respective phosphoramidites or H-phosphonates are used as said nucleotide precursors.
Claim 33
Dependent Claim
21 words
The method of claim 1, wherein said method further comprises incorporating into said RNA chain at least one modified internucleotidic linkage.
Claim 34
Dependent Claim
14 words
The method of claim 31, wherein said modified intemucleotidic linkage is a phosphorothioate group.
Claim 35
Dependent Claim
27 words
The method of claim 1, wherein said synthesis of said RNA chain is carried out by transcription from a nucleic acid template by a nucleic acid polymerase.
Claim 36
Dependent Claim
13 words
The method of claim 1, wherein said RNA chain is a hairpin RNA.
Claim 37
Dependent Claim
13 words
The method of claim 1, wherein said RNA chain is a hammerhead ribozyme.
Claim 38
Dependent Claim
33 words
The method of claim 36, wherein said hammerhead ribozyme comprises a sequence 5'--C--U a --G--A--U a --G--3', wherein each U a represents a uracil modified at said 2'-position with a said modifier group.
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