Upgrade to enjoy subscriber-only features like email alerts and data export. See the Plans and Pricing
|Title:||High affinity nucleic acid ligands containing modified nucleotides|
|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'-NH.sub.2 modified RNA ligands to thrombin are described.|
|Inventor(s):||Pieken; Wolfgang (Longmont, CO), Tasset; Diane (Boulder, CO), Janjic; Nebojsa (Lafayette, CO), Gold; Larry (Boulder, CO), Kirschenheuter; Gary P. (Arvada, CO)|
|Assignee:||NeXstar Pharmaceuticals, Inc. (Boulder, CO)|
|Filing Date:||Apr 27, 1995|
|Claims:||1. 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.
2. A method for claim 1 further comprising
e) repeating steps c) and d).
3. 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.
4. 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.
5. 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.
6. 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.
7. 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).
8. A nucleic acid ligand to thrombin identified according to the method of claim 1.
9. 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.
10. 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.
Drugs may be covered by multiple patents or regulatory protections. All trademarks and applicant names are the property of their respective owners or licensors. Although great care is taken in the proper and correct provision of this service, thinkBiotech LLC does not accept any responsibility for possible consequences of errors or omissions in the provided data. The data presented herein is for information purposes only. There is no warranty that the data contained herein is error free. thinkBiotech performs no independent verifification of facts as provided by public sources nor are attempts made to provide legal or investing advice. Any reliance on data provided herein is done solely at the discretion of the user. Users of this service are advised to seek professional advice and independent confirmation before considering acting on any of the provided information. thinkBiotech LLC reserves the right to amend, extend or withdraw any part or all of the offered service without notice.