Abstract: This invention discloses high-affinity oligonucleotide ligands to the thermostable Taq polymerase and Tth polymerase. Specifically, this invention discloses DNA ligands having the ability to bind to the Taq and Tth polymerases and the methods for obtaining such ligands. The ligands are capable of inhibiting polymerases at ambient temperatures.
Abstract: This invention pertains to methods of determining the presence and/or quantity of homocysteine in a sample containing other thiol-containing compounds. The methods of this invention involve modifying the homocysteine to facilitate the separation of homocysteine from cysteine. An assay protocol comprises adjusting the conditions of the sample suspected of containing homocysteine so that homocysteine forms homocysteine thiolactone, separating the homocysteine thiolactone from free thiol-containing compounds present in the sample, including cysteine, reconverting the homocysteine thiolactone to homocysteine, and determining the presence and/or quantity of homocysteine in the sample by conventional means.
Abstract: This invention discloses high-affinity oligonucleotide ligands to complex tissue targets, specifically nucleic acid ligands having the ability to bind to complex tissue targets, and the methods for obtaining such ligands. Tissue targets comprise cells, subcellular components, aggregates or cells, collections of cells, and higher ordered structures. Specifically, nucleic acid ligands to red blood cells ghosts, endothelia of the blood brain and CSF-blood barriers, glioblastomas, and lymphomas are described.
Abstract: This invention discloses a method for preparing a therapeutic or diagnostic complex comprised of a nucleic acid ligand and a lipophilic compound or non-immunogenic, high molecular weight compound by identifying a nucleic acid ligand by SELEX methodology and associating the nucleic acid ligand with a lipophilic compound or a non-immunogenic, high molecular weight compound. The invention further discloses complexes comprising one or more nucleic acid ligands in association with a lipophilic compound or non-immunogenic, high molecular weight compound.
Type:
Grant
Filed:
May 4, 1995
Date of Patent:
January 4, 2000
Assignee:
NeXstar Pharmaceuticals, Inc.
Inventors:
Larry Gold, Paul G. Schmidt, Nebojsa Janjic
Abstract: This invention discloses high-affinity oligonucleotide ligands to lectins, specifically nucleic acid ligands having the ability to bind to the lectins, wheat germ agglutinin, L-selectin, E-selectin and P-selectin. Also disclosed are the methods for obtaining such ligands.
Type:
Grant
Filed:
June 7, 1995
Date of Patent:
December 14, 1999
Assignee:
NeXstar Pharmaceuticals, Inc.
Inventors:
David H. Parma, Brian J. Hicke, Philippe Bridonneau, Larry Gold
Abstract: A method for identifying nucleic acid ligands to target molecules using the SELEX procedure wherein the candidate nucleic acids contain photoreactive groups and nucleic acid ligands identified thereby are claimed. The complexes of increased affinity nucleic acids and target molecules formed in the procedure are crosslinked by irradiation to facilitate separation from unbound nucleic acids. In other methods partitioning of high and low affinity nucleic acids is facilitated by primer extension steps as shown in the figure in which chain termination nucleotides, digestion resistant nucleotides or nucleotides that allow retention of the cDNA product on an affinity matrix arc differentially incorporated into the cDNA products of either the high or low affinity nucleic acids and the cDNA products are treated accordingly to amplification, enzymatic or chemical digestion or by contact with an affinity matrix.
Type:
Grant
Filed:
June 8, 1998
Date of Patent:
December 14, 1999
Assignee:
NeXstar Pharmaceuticals, Inc.
Inventors:
Larry Gold, Michael Willis, Tad Koch, Steven Ringquist, Kirk Jensen, Brent Atkinson
Abstract: This application provides methods for identifying nucleic acid ligands capable of covalently interacting with targets of interest. The nucleic acids can be associated with various functional units. The method also allows for the identification of nucleic acids that have facilitating activities as measured by their ability to facilitate formation of a covalent bond between the nucleic acid, including its associated functional unit, and its target.
Type:
Grant
Filed:
August 21, 1997
Date of Patent:
December 7, 1999
Assignee:
NeXstar Pharmaceuticals, Inc.
Inventors:
Larry Gold, Bruce Eaton, Drew Smith, Matthew Wecker, Kirk Jensen
Abstract: The presence of a target in a test mixture or the concentration of such target can be determined by a method relying on fluorescence emission measurement. Such method utilizes a nucleic acid ligand to the target and a ligand beacon comprising: i) a nucleic acid sequence complimentary to at least a portion of said nucleic acid ligand, ii) a fluorescent group, and iii) a fluorescence-modifying group. The emission profile of the ligand beacon is altered based on the presence and/or concentration change of the target in the test mixture.
Abstract: Methods are described for the identification and preparation of high-affinity nucleic acid ligands to cytokines. Included in the invention are specific nucleic acid ligands to IFN-gamma, IL-4, IL-10, TNF-alpha, and RANTES.
Type:
Grant
Filed:
June 7, 1995
Date of Patent:
October 26, 1999
Assignee:
NeXstar Pharmaceuticals, Inc.
Inventors:
Diane Tasset, Nikos Pagratis, Sumedha Jayasena, Larry Gold
Abstract: A novel composition and method for solubilizing amphiphilic drugs in a small amount of organic solvent for use in improved liposomes is disclosed. A phosphatidylglycerol is acidified in a small amount of organic solvent. The amphiphilic drug, such as Amphotericin B, suspended in organic solvent is then added to the acidified phosphatidylycerol and a soluble complex is formed between the phosphatidylglycerol and the amphiphilic drug. When the liposome composition incorporating the soluble complex is hydrated, the final pH of the hydrating aqueous buffer is carefully controlled. The Amphotericin B liposomes formed have markedly reduced toxicity.
Type:
Grant
Filed:
June 6, 1995
Date of Patent:
October 12, 1999
Assignee:
NeXstar Pharmaceuticals, Inc.
Inventors:
Richard T. Proffitt, Jill Adler-Moore, Su-Ming Chiang
Abstract: This application provides methods for identifying nucleic acid ligands capable of covalently interacting with targets of interest. The nucleic acids can be associated with various functional units. The method also allows for the identification of nucleic acids that have facilitating activities as measured by their ability to facilitate formation of a covalent bond between the nucleic acid, including its associated functional unit, and its target.
Type:
Grant
Filed:
June 5, 1995
Date of Patent:
October 5, 1999
Assignee:
NeXstar Pharmaceuticals, Inc.
Inventors:
Larry Gold, Bruce Eaton, Drew Smith, Matthew Wecker, Kirk Jensen
Abstract: 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.
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.
Type:
Grant
Filed:
August 25, 1997
Date of Patent:
September 28, 1999
Assignee:
NeXstar Pharmaceuticals, Inc.
Inventors:
Wolfgang Pieken, Diane Tasset, Nebojsa Janjic, Larry Gold, Gary P. Kirschenheuter
Abstract: A liposomal aminoglycoside formulation comprising a neutral lipid, a negatively charged lipid and a sterol. The formulation contains unilamellar vesicles having an average size below 100 nm. A process of making liposomes containing an aminoglycoside is provided. A method is also provide for the treatment of drug susceptible and drug resistant bacteria.
Type:
Grant
Filed:
May 21, 1998
Date of Patent:
September 28, 1999
Assignee:
NeXstar Pharmaceuticals, Inc.
Inventors:
Evan M. Hersch, Eskild A. Petersen, Richard T. Proffitt, Kevin R. Bracken, Su-Ming Chiang
Abstract: This invention discloses a method for the preparation modified nucleosides and nucleotides using a palladium catalyst, a nucleophile and carbon monoxide.
Abstract: A liposome formulation containing a Vinca Alkaloid and an anion in an aqueous phase of the liposome. The liposomes also comprise distearoylphosphatidyl choline, cholesterol and disteraroylphosphatidylglycerol. A method for enhancing the efficacy and tumor targeting properties of liposomal vinca alkaloid formulations containing unilamellar vesicles.
Abstract: High-affinity oligonucleotide ligands to the thermostable Taq polymerase and Tth polymerase are disclosed. Specifically disclosed are DNA ligands having the ability to bind to the Taq and Tth polymerases and the methods for obtaining such ligands. The ligands are capable of inhibiting polymerases at ambient temperatures.
Abstract: This invention discloses an improved method for the sequential solution phase synthesis of oligonucleotides and peptides. The method lends itself to automation and is ideally suited for large scale manufacture oligonucleotides with high efficiency.
Abstract: Formulations consisting of phospholipid small unilamellar particles encapsulating polyene antifungal antibiotics and methods for using such compositions to treat systemic fungal infections are described. In a preferred embodiment, the particles are in the form of vesicles which comprise a polyene antifungal antibiotic, preferably amphotericin B and/or nystatin, egg phosphatidylcholine and cholesterol, preferably in the molar ratio of about 0.2 (AMB):2(PL):1(CHOL). These vesicles may also have an amine modified surface. The vesicles are suspended in a low isonic strength saccharide/tris solution at a pH of from about 6.0 to about 8.0 and may be administered to deliver the antifungal antibiotic to treat systemic fungal infections.
Type:
Grant
Filed:
June 6, 1995
Date of Patent:
February 23, 1999
Assignee:
NeXstar Pharmaceuticals, Inc.
Inventors:
Jill Adler-Moore, Ronald C. Gamble, Richard T. Proffitt
Abstract: This invention discloses novel detection methods for determining the presence of a target compound in a substance using nucleic acid ligands as detection molecules. Specifically, the substance is bound to a solid support matrix, such as those used in blot procedures, and detection of the target molecule is accomplished using the affinity and specificity of nucleic acid ligands to the target molecule. The method utilized herein for identifying and preparing said nucleic acid ligands is called SELEX. The method of the present invention is additionally useful to isolate the target compounds from the various substances.
Type:
Grant
Filed:
April 5, 1996
Date of Patent:
February 23, 1999
Assignee:
NeXstar Pharmaceuticals, Inc.
Inventors:
Dan Drolet, Sumedha D. Jayasena, Larry Gold