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US5660985A
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High affinity nucleic acid ligands containing modified nucleotides
Assignee: Gilead Sciences Inc
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Filing: 1995-04-27
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Granted: 1997-08-26
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Expires: 2015-04-27
Abstract
Abstract Nucleic acid ligands containing modified nucleotides are described as are methods for producing such oligonucleotides. Such ligands enrich the chemical diversity of the candidate mixture for the SELEX process. Specific examples are provided of nucleic acids containing nucleotides modified at the 2'- and 5-position. Specific 2-OH and 2'-NH2 modified RNA ligands to thrombin are described.
Inventors
- 👤 Wolfgang Pieken
- 👤 Diane Tasset
- 👤 Nebojsa Janjic
- 👤 Larry Gold
- 👤 Gary P. Kirschenheuter
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 16:56
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/115; A; A61; A61K; A61K47/00; A61K47/50; A61K47/51; A61K47/54; A61K47/547; A61K47/549; B; B82; B82Y; B82Y5/00; C07; C07H; C07H19/00; C07H19/02; C07H19/04; C07H19/06; C07H19/10; C07H21/00; C12N15/10; C12N15/1034; C12N15/1037; C12N15/1048; C12Q; C12Q1/00; C12Q1/68; C12Q1/6811; C40; C40B; C40B40/00; C40B40/02; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/569; G01N33/56983; G01N33/56988; G01N33/74; G01N33/76; G01N2333/00; G01N2333/005; G01N2333/08; G01N2333/15; G01N2333/155; G01N2333/16; G01N2333/163; G01N2333/435; G01N2333/475; G01N2333/50; G01N2333/503; G01N2333/575; G01N2333/62; G01N2333/81; G01N2333/8107; G01N2333/811; G01N2333/8121; G01N2333/8125; G01N2333/90; G01N2333/914; G01N2333/948; G01N2333/95; G01N2333/964; G01N2333/96425; G01N2333/96427; G01N2333/9643; G01N2333/96433; G01N2333/96441; G01N2333/96455; G01N2333/966; G01N2333/972; G01N2333/9726; G01N2333/974; G01N2333/976
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
123 words
A method for identifying nucleic acid ligands comprising a modified nucleotide to a target molecule comprising: a) preparing a candidate mixture of single-stranded nucleic acids wherein each nucleic acid of said candidate mixture comprises a modified nucleotide selected from the group consisting of a 5-position modified pyrimidine, a 2'-position modified pyrimidine, an 8 -position modified purine, and a 2'-position modified purine; b) contacting the candidate mixture with said target molecule; c) partitioning the nucleic acids having an increased affinity to the target molecule relative to the candidate mixture from the remainder of the candidate mixture; and d) amplifying the increased affinity nucleic acids, in vitro, to yield a ligand-enriched mixture of nucleic acids, whereby nucleic acid ligands of the target molecule are identified.
Claim 2
Independent Claim
123 words
A method for identifying nucleic acid ligands comprising a modified nucleotide to a target molecule comprising: a) preparing a candidate mixture of single-stranded nucleic acids wherein each nucleic acid of said candidate mixture comprises a modified nucleotide selected from the group consisting of a 5-position modified pyrimidine, a 2'-position modified pyrimidine, an 8 -position modified purine, and a 2'-position modified purine; b) contacting the candidate mixture with said target molecule; c) partitioning the nucleic acids having an increased affinity to the target molecule relative to the candidate mixture from the remainder of the candidate mixture; and d) amplifying the increased affinity nucleic acids, in vitro, to yield a ligand-enriched mixture of nucleic acids, whereby nucleic acid ligands of the target molecule are identified.
Claim 3
Dependent Claim
13 words
A method for claim 1 further comprising e) repeating steps c) and d).
Claim 4
Dependent Claim
22 words
The method of claim 1 wherein said single-stranded nucleic acids are selected from the group consisting of single-stranded DNA and single-stranded RNA.
Claim 5
Dependent Claim
25 words
The method of claim 1 wherein said modified nucleotide is selected from the group consisting of a 2'-position modified pyrimidine and a 5-position modified pyrimidine.
Claim 6
Dependent Claim
23 words
The method of claim 1 wherein said modified nucleotide is selected from the group consisting of a 2'-amino pyrimidine and a 2'-amino purine.
Claim 7
Dependent Claim
23 words
The method of claim 1 wherein said modified nueleotide is selected from the group consisting of a 2'-fluoro pyrimidine and a 2'-fluoro purine.
Claim 8
Dependent Claim
32 words
The method of claim 4 wherein said 5-position modified pyrimidine is selected from the group consisting of 5-(3-amino alkyl) uridine triphosphate (5-AA-UTP), 5-fluorescein-12-uridine triphosphate (5-F-12-UTP), 5-digoxygenin-11-uridine triphosphate (5-Dig-11-UTP), and 5-bromouridine triphosphate (5-Br-UTP).
Claim 9
Dependent Claim
14 words
A nucleic acid ligand to thrombin identified according to the method of claim 1.
Claim 10
Dependent Claim
32 words
The nucleic acid ligand of claim 8 wherein said nucleic acid ligand is comprised of 2'-amino pyrimidines, and wherein said ligand is selected from the group consisting of SEQ ID NOS: 20-39.
Claim 11
Dependent Claim
23 words
The method of claim 1 wherein said modified nucleotide is selected from the group consisting of a 2'-methoxy pyrimidine and a 2'-methoxy purine.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2000-08-25 | 2001-02-24 | 2001-08-26 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2004-08-25 | 2005-02-24 | 2005-08-26 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2008-08-25 | 2009-02-24 | 2009-08-26 | $7700.00 | Window Closed / Lapsed |