Nexstar Pharmaceuticals, Inc.

Cortech, Inc.

Gilead Sciences, Inc.

The Johns Hopkins University

Gary P Kirschenheuter

Larry M Gold

John C Cheronis

Bruce Eaton

Drew Smith

Nebojsa Janjic

Sumedha D Jayasena

Wolfgang A Pieken

Diane Tasset

Greg Biesecker

Lyle W Spruce

James K Blodgett

Howard M Dintzis

Raymond T Cunningham

Renee Z Dintzis

Gregory Biesecker

Jozef Oleksyszyn

Sumeda Jayasena

Ren&eacute;e Z Dintzis

A method is described for generating blended nucleic acid ligands containing non-nucleic acid functional units. Specifically, a SELEX identified RNA ligand to the integrin gpIIbIIIa is conjugated to the peptide Gly-Arg-Gly-Asp-Thr-Pro (SEQ ID NO:1). This blended RNA ligand inhibits the biological activity of gpIIbIIIa with high specificity. Also described is a single-stranded DNA ligand to elastase coupled to N-methoxysuccinyl-Ala-Ala-Pro-Val-chloromethyl ketone (SEQ ID NO:2). This elastase blended nucleic acid ligand inhibits the biological activity of elastase.

Systematic evolution of ligands by exponential enrichment: blended SELEX

Modified nucleosides and methods of making and using the nucleosides are disclosed. The compounds can be prepared by reacting nucleoside starting materials that contain a suitable leaving group at one or more of the carbon atoms in the purine or pyrimidine ring, with a vinylstannane, carbon monoxide, and a palladium catalyst to provide 1-ene-3-one intermediates. These intermediates are then reacted with suitably functionalized primary or secondary amines via a Michael reaction. When the intermediate is a 5-position modified pyrimidine ring, and the amine contains a second hydrogen, it can do a second Michael reaction with the ene-one or the ene-imine in the pyrimidine ring. Appropriate modification of the amine reactant can yield products with various bioactivities. The nucleosides can be used therapeutically as anti-cancer, anti-bacterial or anti-viral drugs. The nucleosides can also be used for diagnostic applications, for example, by incorporating a radiolabel or fluorescent label into the molecule. The nucleosides can be used to prepare oligonucleotides for use in various applications, either alone or in combination with other modified nucleosides and/or naturally occurring nucleosides.

Urea nucleosides as therapeutic and diagnostic agents

Systematic evolution of ligands by exponential enrichment: blended selex

The invention features a non-immunogenic construct for reducing a non-primary antibody response to an epitope of a T-dependent antigen. The construct is free of high molecular weight immunostimulatory molecules and contains copies of the epitope which bind to a B cell membrane immunoglobulin receptor specific for the epitope but fail to form an imnunon.

Therapeutic suppression of specific immune response by administration of oligomeric forms of antigen of controlled chemistry

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&prime;- and 5-position. Specific 2-OH and 2&prime;-NH,modified RNA ligands to thrombin are described.

High affinity nucleic acid ligands containing modified nucleotides

A method is described for generating blended nucleic acid ligands containing non-nucleic acid functional units. Specifically, a SELEX identified RNA ligand to the integrin gpIIbIIIa is conjugated to the peptide Gly-Arg-Gly-Asp-Thr-Pro (SEQ ID NO:1). This blended RNA ligand inhibits the biological activity of gpIIbIIIa with high specificity. Also described is a single-stranded DNA ligand to elastase coupled to N-methoxysuccinyl-Ala-Ala-Pro-Val-chloromethyl (SEQ ID NO:3) ketone. This elastase blended nucleic acid ligand inhibits the biological activity of elastase.

Systematic evolution of ligands exponential enrichment: blended selex

The invention features a method for reducing an undesired antibody response in a mammal by administering to the mammal a non-immunogenic construct which is free of high molecular weight immunostimulatory molecules. The construct which contains at least two copies of a B cell membrane immunoglobulin receptor epitope bound to a pharmaceutically acceptable non-immunogenic carrier directly binds to B cell membrane immunoglobulin receptors but fails to form an immunon.

Therapeutic suppression of specific immune responses by administration

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.

Pyrimidine nucleosides as therapeutic and diagnostic agents

A method is described for generating blended nucleic acid ligands containing non-nucleic acid functional units. Specifically, a SELEX identified RNA ligand to the integrin gpIIbIIIa is conjugated to the peptide Gly-Arg-Gly-Asp-Thr-Pro (SEQ ID NO:1). This blended RNA ligand inhibits the biological activity of gpIIbIIIa with high specificity. Also described is a single-stranded DNA ligand to elastase coupled to N-methoxysuccinyl-Ala-Ala-Pro-Val-chloromethyl (SEQ ID NO:2) ketone. This elastase blended nucleic acid ligand inhibits the biological activity of elastase.

2-Phenylalkanoate esters which are useful as inhibitors of human leukocyte elastase.

Oxidant sensitive and insensitive aromatic esters as inhibitors of human

2-Heteroaromatic alkanoate esters which are useful as inhibitors of human leukocyate or neutrophil elastase.

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Arvada, CO, United States of America

Gary P Kirschenheuter