WO2000012530A1 - CLIVAGE DIRIGE SUR SITE dU DE CONJUGUES COVALENTS - Google Patents

CLIVAGE DIRIGE SUR SITE dU DE CONJUGUES COVALENTS Download PDF

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Publication number
WO2000012530A1
WO2000012530A1 PCT/US1999/018091 US9918091W WO0012530A1 WO 2000012530 A1 WO2000012530 A1 WO 2000012530A1 US 9918091 W US9918091 W US 9918091W WO 0012530 A1 WO0012530 A1 WO 0012530A1
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Prior art keywords
nucleic acid
target
covalent conjugate
ligand
nucleic acids
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PCT/US1999/018091
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English (en)
Inventor
Russ Lehrman
Stephanie Niewlandt
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Nexstar Pharmaceuticals, Inc.
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Publication date
Application filed by Nexstar Pharmaceuticals, Inc. filed Critical Nexstar Pharmaceuticals, Inc.
Priority to AU53462/99A priority Critical patent/AU5346299A/en
Publication of WO2000012530A1 publication Critical patent/WO2000012530A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/115—Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide

Definitions

  • This invention relates to covalent conjugates and prodrug forms of nucleic acids and their use in the purification, pharmaceutical and diagnostic arts. Specifically, this invention
  • This invention describes a method for identifying covalent conjugates comprising a nucleic acid component covalently bonded to a separable unit via a deoxyuridine. This invention further describes a method of analyzing the nucleic acid component of a covalent conjugate by treating the
  • the covalent conjugates of the present invention as therapeutic and diagnostic agents.
  • oligonucleotides are used to bind to certain coding regions in an organism to prevent the expression of proteins or to block various cell functions. Further, the discovery of the
  • nucleic acids had primarily an informational
  • SELEX is a method for the in vitro evolution of
  • nucleic acid molecules with highly specific binding to target molecules and is described in
  • nucleic acid ligands also referred to as nucleic acid ligands
  • aptamers each ligand having a unique sequence, and which has
  • SELEX-identified nucleic acid ligand is a specific ligand of a given target compound or
  • SELEX is based on the unique insight that nucleic acids have sufficient capacity
  • Molecules of any size or composition can serve as
  • the SELEX method involves selection from a mixture of candidate oligonucleotides
  • the SELEX method includes steps of contacting the mixture with the
  • nucleic acids which have bound specifically to target molecules, dissociating the nucleic acids
  • binding binding, partitioning, dissociating and amplifying through as many cycles as desired to yield
  • the SELEX method demonstrates that nucleic acids as chemical compounds can
  • nucleic acids could also be used to identify nucleic acids which can facilitate any chosen reaction in a
  • nucleic acid ligands can be identified for any given target.
  • the SELEX method encompasses the identification of high-affinity nucleic acid
  • modifications include chemical substitutions at the ribose and/or phosphate and/or base
  • SELEX-identified nucleic acid ligands containing modified nucleotides are
  • the SELEX method encompasses combining selected oligonucleotides with other
  • the SELEX method encompasses complexes of oligonucleotides. United States
  • nucleic acid ligand comprised of a nucleic acid ligand and a lipophilic compound or a non-immunogenic, high
  • Antisense oligonucleotides therefore, are
  • the drug to a carrier capable of transporting and/or targeting the drug from the site of
  • linear polymers (2) cells; (3) three dimensional systems (e.g., liposomes); and (4)
  • conjugate is in the target area, the linkage between the drug and the carrier is cleaved
  • Such carriers include proteins, which are
  • glycosidases or vinyl polymers which contain hydrolytically labile ester bonds.
  • al. describe a method for chemically modifying antisense oligonucleotides to enhance entry of the drug into a cell by conjugating an oligonucleotide via a disulfide bond to a transport
  • Biopolymers therefore can be constructed to possess domains that have
  • hybrids have been prepared in which DNA is linked to targeting, cleaving or reporter groups
  • oligonucleotides to lipophilic compounds or non-immunogenic, high molecular weight
  • lipophilic compound covalently attached to an antisense oligonucleotide through a
  • Linear polymers are typically covalently linked to a drug via a hydrolyzable bond
  • ester bonds can be quickly hydrolyzed in biological fluids by
  • ester linkage can increase the instability of such compounds due to hydrolysis of the ester bond, which can occur during
  • DNA glycosylases initiate DNA repair pathways in many organisms by excising
  • Uracil DNA glycosylases (UDG), purified
  • UDG eliminates uracil from
  • PCRs polymerase chain reactions
  • nucleases are known which specifically cleave single-stranded RNA at individual
  • Type II restriction endonucleases have been well characterized.
  • Fokl which is known to specifically recognize a pentanucleotide sequence in
  • the present invention provides covalent conjugates comprising one or more nucleic acids
  • nucleic acid ligand In the preferred embodiment, the nucleic acid ligand is identified according to the SELEX method. In one embodiment, the nucleic acid component is single-stranded
  • the nucleic acid component is RNA. In embodiments in
  • RNA nucleic acid component is a nucleic acid wherein all nucleotides are 2'-OH
  • the covalent conjugate contains one or more DNA residues between the dU and
  • RNA nucleic acid component the RNA nucleic acid component
  • the separable unit is a nucleic acid.
  • the separable unit is a non-immunogenic, high molecular
  • PEG polyethylene glycol
  • the separable unit is a lipophilic compound.
  • compound is cholesterol, a dialkyl glycerol or a diacyl glycerol.
  • the present invention provides means for the site directed
  • the nucleic acid is site-specifically cleaved from the attached separable unit
  • this invention provides a method for targeting a therapeutic
  • the therapeutic or diagnostic agent is administered to the patient.
  • the therapeutic or diagnostic agent is administered to the patient.
  • the covalent bond is the nucleic acid component of the covalent conjugate.
  • the covalent bond is the nucleic acid component of the covalent conjugate.
  • conjugate comprises a nucleic acid component which is inactive when linked to the
  • the present invention further provides a method for analyzing the structure of a
  • nucleic acid component of a covalent conjugate nucleic acid component of a covalent conjugate
  • the present invention further provides means for analyzing a nucleic acid
  • This invention further provides covalent conjugates wherein the nucleic acid
  • component of the covalent conjugate is an intracellular agent and the separable unit is
  • the separable unit is cleaved by the intracellular UDG, effecting the release of the
  • FIG 1 shows the anion exchange HPLC trace of the digestion of 5K-PEG-dU303
  • FIG. 1 shows the anion exchange HPLC trace of the digestion of
  • the present invention provides covalent conjugates comprising one or more nucleic acids
  • the covalent conjugates of the present invention have the formula
  • A-B-C wherein A is RNA, single-stranded DNA or double stranded DNA, B is one or more
  • deoxyuridines and C is a separable unit, wherein both A and C are covalently bonded to B.
  • A is RNA comprising all 2'-OH nucleotides
  • the nucleic acid component of the covalent bond in one embodiment of the invention, the nucleic acid component of the covalent bond
  • the nucleic acid component is N-stranded DNA.
  • the nucleic acid component is a nucleic acid ligand. In another embodiment,
  • the nucleic acid component is a nucleic acid ligand identified by the SELEX
  • the separable unit is bound to a hydroxyl group at the 5' or 3'
  • 2'-modified pyrimidines for example, 2'-F, 2'-NH 2 or 2'-OMe.
  • the separable unit of the covalent conjugate is
  • a lipophilic compound such as cholesterol, a dialkyl glycerol, a diacyl glycerol, or a non-
  • the separable unit is covalently attached to the dU.
  • the separable unit is covalently attached to the dU through a linker.
  • conjugate comprises a linker between the separable unit and the dU
  • the linker comprises a reactive group.
  • the linker comprises a primary
  • nucleic acid ligand as used herein is a nucleic acid having a desirable action on a
  • a desirable action includes, but is not limited to, binding of the target, catalytically
  • the action is specific binding affinity for a target molecule, such target
  • nucleic acid ligand is not a nucleic acid ligand
  • nucleic acid having the known physiological function of being bound by the target
  • the nucleic acid ligand is a non-naturally occurring nucleic
  • nucleic acid ligands are identified by
  • Nucleic acid ligands include nucleic acids that are identified
  • nucleic acid ligand being a ligand of a given
  • the method comprising a) contacting the candidate mixture with the target,
  • mixture may be partitioned from the remainder of the candidate mixture; b) partitioning the
  • Candidate Mixture is a mixture of nucleic acids of differing sequence from which
  • the source of a candidate mixture can be from naturally-reacted ligands.
  • nucleic acids or fragments thereof chemically synthesized nucleic acids
  • each nucleic acid has fixed sequences to
  • Nucleic acid means either DNA, RNA, single-stranded or double-stranded and any
  • nucleic acid ligand as a whole.
  • modifications include, but are not limited to, 2'-
  • Non-immunogenic, high molecular weight compound is a compound of
  • An immunogenic response is one that induces the organism to produce antibody proteins.
  • non-immunogenic, high molecular weight compounds examples include polyethylene
  • PEG glycol
  • polysaccharides such as dextran
  • polypeptides such as albumin
  • the non-immunogenic is selected from the non-immunogenic
  • high molecular weight compound can also be a nucleic acid ligand.
  • Lipophilic compounds are compounds which have the propensity to associate with
  • phospholipids diacyl glycerols and dialkyl glycerols are examples of lipophilic compounds.
  • "SELEX" methodology involves the combination of selection of nucleic acid ligands
  • the SELEX methodology can be employed to obtain a
  • nucleic acid ligand to a desirable target.
  • Target means any compound of interest for which a ligand is desired.
  • molecule can be a protein, peptide, carbohydrate, polysaccharide, glycoprotein, hormone,
  • receptor antigen, antibody, virus, substrate, metabolite, cell, tissue, transition state analog
  • cofactor cofactor, inhibitor, drug, dye, nutrient, growth factor, etc., without limitation.
  • Covalent conjugate as used herein describes the molecular entity having the
  • A is RNA, single-stranded DNA or double stranded DNA
  • B is one
  • deoxyuridines and C is a separable unit, wherein both A and C are covalently
  • A is RNA comprising all 2'-OH nucleotides
  • Synthetic unit includes oligonucleotides, lipophilic compounds,
  • non-immunogenic, high molecular weight compounds such as nucleic acids, proteins, carbohydrates, organic compounds that chelate metals, detectable moieties such as
  • Uracil DNA glycosylase (UDG), a term of art, is a DNA glycosylase which
  • UDG eliminates uracil from DNA by cleaving the N-glycosidic bond between the base
  • Deoxyuridine is a nucleoside residue which can arise as a result of either
  • Linker is a molecular entity that connects two or more molecular entities through a
  • linkers include an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound that can be attached to an organic compound
  • oligonucleotide and has a chemically reactive group such as a free amino, sulfhydryl, diene,
  • Covalent bond is the chemical bond formed by the sharing of electrons.
  • Therapeutic Agent means a compound which is used in the treatment of diseases
  • covalent conjugate can be for targeting purposes and the separable unit is the therapeutic
  • nucleic acid component of the covalent conjugate is the nucleic acid component of the covalent conjugate
  • Diagnostic Agent means a covalent conjugate which can be used for detecting the
  • target molecule is mediated by its binding to a nucleic acid component of a covalent
  • the covalent conjugate can be labeled, for
  • radiolabeled to allow qualitative or quantitative detection.
  • the present invention provides covalent conjugates comprising one or more nucleic acids
  • nucleic acid component and the separable unit provides means for site-directed cleavage
  • nucleic acid component to a separable unit through a dU imparts improved stability to the
  • the present invention allows for the utilization of base-sensitive RNA as the nucleic acid component of the covalent conjugate. Therefore, the separable unit enhances certain
  • the present invention provides methods for the isolation of the nucleic acid cleaved
  • nucleic acid conjugate comprising a nucleic acid component coupled to a PEG, the nucleic acid is
  • conjugate and then the covalent conjugate product is purified, usually by extraction into an organic solvent
  • a nucleic acid can be any nucleic acid following purification.
  • a nucleic acid can be any nucleic acid following purification.
  • the covalent conjugate can then be easily purified, and
  • nucleic acid component of the covalent conjugate can be cleaved
  • nucleic acid can be analyzed.
  • the method of the present invention further provides covalent conjugates
  • the covalent conjugate is administered to a patient and the nucleic acid
  • nucleic acid component of a covalent conjugate is active
  • the separable unit when in a hairpin or stem-loop or pseudoknot conformation, and the separable unit is a
  • nucleic acid sequence which is complementary to a portion of the nucleic acid and thus
  • covalent conjugates of the present invention may be used as therapeutic or
  • the present invention provides means for treating a disease using
  • conjugate is administered to a patient for diagnostic or therapeutic purposes, the covalent
  • conjugate comprises a nucleic acid component which is inactive while covalently bonded to
  • nucleic acid component of the covalent conjugate is an
  • the separable unit is a carrier capable of transporting the nucleic
  • nucleic acid component activates upon cleavage from the covalent conjugate by the
  • a covalent conjugate is to be used as a therapeutic agent, it may be desirable to
  • covalent conjugates described herein are particularly useful as in vivo or in vitro
  • the covalent conjugates of the present invention are used.
  • diagnostic agents for detecting the presence or absence of a target molecule in a sample.
  • the covalent conjugate comprises a nucleic acid ligand to a desired
  • the separable unit comprises a detectable moiety such as a biotin, an enzyme or a
  • binding or interaction can be
  • This method can be useful as a diagnostic tool, for example
  • conjugate comprising a specific oligonucleotide that selectively binds to or interacts with
  • oligonucleotides in the phosphodiester form can be quickly degraded in body
  • ligand can be made to increase the in vivo stability of the nucleic acid ligand or to enhance
  • nucleic acid ligand Modification of the nucleic acid
  • ligands contemplated in this invention include, but are not limited to, those which provide
  • nucleic acid ligands are derived by the SELEX method, the modifications
  • Pre-SELEX modifications yield nucleic acid
  • nucleic acid ligands of the invention are 5' and 3' phosphorothioate
  • nucleotides is preferred.
  • nucleic acid ligands of the present invention are N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl
  • the SELEX process provides a class of products which are nucleic acid molecules
  • Target molecules are preferably proteins, but can also include among others carbohydrates, peptidoglycans and a variety of small molecules.
  • SELEX methodology can also be used to target biological structures, such as cell surfaces or
  • the SELEX process may be defined by the following series of
  • a candidate mixture of nucleic acids of differing sequence is prepared.
  • candidate mixture generally includes regions of fixed sequences (i.e., each of the members
  • the fixed sequence regions are selected either: (a) to assist in the
  • the randomized sequences can be totally randomized (i.e., the
  • the probability of finding a base at any location can be selected at any level between 0
  • mixture can be considered as forming nucleic acid-target pairs between the target and those
  • nucleic acids having the strongest affinity for the target having the strongest affinity for the target.
  • affinity for the target are then amplified to create a new candidate mixture that is enriched in
  • nucleic acids having a relatively higher affinity for the target are provided.
  • candidate mixture contains fewer and fewer unique sequences, and the average degree of
  • nucleic acids representing those nucleic acids from the original candidate mixture having
  • nucleic acid ligands containing photoreactive groups capable of binding and/or
  • nucleic acid ligands able to discriminate between closely related molecules termed
  • the SELEX method encompasses the identification of high-affinity nucleic acid
  • SELEX-identified nucleic acid ligands containing modified nucleotides are
  • nucleotide derivatives chemically modified at the 5- and 2'-positions of
  • the SELEX method encompasses combining selected oligonucleotides with other
  • SELEX identifies nucleic acid ligands that are able to bind targets with high affinity
  • the nucleic acid ligand (1) binds to the target in a manner capable of achieving
  • the desired effect on the target (2) be as small as possible to obtain the desired effect; (3) be
  • the nucleic acid ligand has the highest possible affinity to the target.
  • compositions of the covalent conjugates may be administered
  • iontophoresis or suppositories are also envisioned.
  • One preferred carrier is physiological
  • such a carrier may be either aqueous or non-aqueous in nature.
  • the carrier may be either aqueous or non-aqueous in nature.
  • the carrier may be either aqueous or non-aqueous in nature.
  • the carrier may be either aqueous or non-aqueous in nature.
  • the carrier may be either aqueous or non-aqueous in nature.
  • the carrier may be either aqueous or non-aqueous in nature.
  • the carrier may
  • the carrier may contain still other pharmacologically-acceptable
  • the therapeutic composition may be stored in sterile
  • vials as a solution, suspension, gel, emulsion, solid, or dehydrated or lyophilized powder.
  • Such formulations may be stored either in a ready to use form or requiring reconstitution
  • covalent conjugates for systemic delivery may be via subcutaneous, intramuscular,
  • UDG may be via subcutaneous, intramuscular or intravenous injection.
  • Oligonucleotides containing dU were synthesized using standard phosphoramidite
  • glycol was conjugated to several oligonucleotides via an amine linker and the resulting
  • conjugated oligonucleotides were purified by organic extraction and liquid chromatographic
  • Example 1 describes the conjugation of
  • NX303 is an L-Selectin ligand
  • AdTdGdG3'3'dTdT-3' (SEQ ID NO:4).
  • Example 3 was performed to investigate the reactivity of UDG on an oligonucleotide
  • Example 5 describes the digestion with UDG of an RNA oligonucleotide having a
  • Example 6 describes the digestion with UDG of an RNA oligonucleotide having a
  • Example 7 describes the digestion with UDG of an L-selectin specific nucleic acid
  • ligand 21918 (NH 2 -C6-dUdTrGrGrArGfUfCfUfUrArGrGfCrArGfCrGfUfUfUfUfCr
  • NX 11702 is described in PCT Application WO 96/40703 , which is
  • Example 8 describes the digestion with UDG of an anti-VEGF nucleic acid ligand
  • This oligonucleotide has been modified from an original nucleic acid ligand
  • VEGF VEGF Nucleic Acid Ligand Complexes
  • NX22022 from plasma compared with the monoexponential clearance of NX31838.
  • phase t 1/2 (283 minutes) for the dU-containing ligand was found to be similar to the overall
  • Uracil DNA Glycosylase was obtained from Giboco Life Technologies (Cat. No.
  • UDG Buffer pH 8.4 20 mM Tris, 50 mM KC1, 5 mM MgCl 2 at pH 8.4.
  • dithiothreitol 1 mM EDTA, 10 mM NaCl.
  • Oligodeoxyribonucleotides were obtained from Operan Technologies (Alameda,
  • Methoxy SPA 5K Peg at the 5' end as follows: Solution (A) was prepared by combining 33
  • Solution (B) was prepared by adding 3.675 mg Methoxy SPA 5,000 MW
  • Chloroform 150 ⁇ L was added, followed by
  • the PEGylated material was purified by reverse phase chromatography using a
  • NXdU303 (NXdU303, NeXstar) (SEQ ID NO:3). NXdU303 was conjugated with Methoxy SPA 5K
  • Solution (A) was prepared by combining 40 ⁇ L (5 ⁇ g/ ⁇ L) of 5'-(5-amino-
  • TdT-3' (5KPEG-dU303) was dried on a speed vacuum overnight.
  • the PEGylated material was purified by reverse phase chromatography using a
  • Acetonitrile gradient 0-10 min: 95% (A)/5% (B) to 80% (A)/20% (B); 10-50 min: 80%
  • oligonucleotide concentration at 90°C.
  • concentration of oligonucleotide in solution at 90°C.
  • GdG3'3'dTdT-3' (SEQ ID NO:4) were 16.15 minutes and 19.63 minutes, respectively
  • oligonucleotide in solution was kept constant at 0.1 ⁇ g/ ⁇ L in UDG buffer (pH 8.4).
  • rArCrGrArCrGrA were 11.5 minutes and 10.9 minutes, respectively.
  • RNA oligonucleotide model with an additional dTdUdT on the 5' end,
  • dTdUdTrArCrGrArCrGrA (Oligo Etc. Inc.) (SEQ ID NO:8) was digested in UDG buffer pH
  • UDG was combined with 5 ⁇ g oligonucleotide and 242 ⁇ L UDG buffer pH 7.8 (Tris) in the
  • RNA oligonucleotide model with dUdT on the 5' end dUdTrArCrGrArCrGrA
  • dTrArCrGrArCrGrA (SEQ ID NO: 11 ) were 18.2 minutes and 18.5 minutes, respectively
  • NX21918 was digested in UDG buffer pH
  • NeXstar was digested with UDG buffer pH 8.4 at 90°C and at 45°C.

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Abstract

Cette invention porte sur un procédé d'identification et de préparation de conjugués covalents comprenant un composant d'acide nucléique lié de manière covalente à une unité séparable par l'intermédiaire d'une désoxyuridine. Cette invention porte également sur des conjugués covalents comprenant un composant d'acide nucléique lié de manière covalente à une unité séparable par l'intermédiaire d'une désoxyuridine. L'invention porte en outre sur un procédé d'analyse d'un composant d'acide nucléique d'un conjugué covalent comprenant un composant d'acide nucléique lié de manière covalente à une unité séparée par l'intermédiaire d'une désoxyuridine.
PCT/US1999/018091 1998-08-26 1999-08-10 CLIVAGE DIRIGE SUR SITE dU DE CONJUGUES COVALENTS WO2000012530A1 (fr)

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Application Number Priority Date Filing Date Title
AU53462/99A AU5346299A (en) 1998-08-26 1999-08-10 Du site-directed cleavage of covalent conjugates

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US14009998A 1998-08-26 1998-08-26
US09/140,099 1998-08-26

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1620450A2 (fr) * 2003-04-13 2006-02-01 Enzon Pharmaceuticals, Inc. Promedicaments oligonucleotidiques polymeres
US9506897B2 (en) 2012-11-16 2016-11-29 Agilent Technologies, Inc. Methods and compositions for improved ion-exchange chromatography
CN108676130A (zh) * 2018-06-06 2018-10-19 河南师范大学 用于药物载体的还原响应型核交联聚合物MPEG-NSASS-Chol胶束的制备

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991014696A1 (fr) * 1990-03-29 1991-10-03 Gilead Sciences, Inc. Conjugues a base de disulfure d'oligonucleotide et d'un agent de transport
WO1993020694A1 (fr) * 1992-04-14 1993-10-28 Signal Pharmaceuticals, Inc. Procede et composition pour l'inhibition de la replication virale
US5660985A (en) * 1990-06-11 1997-08-26 Nexstar Pharmaceuticals, Inc. High affinity nucleic acid ligands containing modified nucleotides
US5723594A (en) * 1995-06-07 1998-03-03 Nexstar Pharmaceuticals, Inc. High affinity PDGF nucleic acid ligands

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991014696A1 (fr) * 1990-03-29 1991-10-03 Gilead Sciences, Inc. Conjugues a base de disulfure d'oligonucleotide et d'un agent de transport
US5660985A (en) * 1990-06-11 1997-08-26 Nexstar Pharmaceuticals, Inc. High affinity nucleic acid ligands containing modified nucleotides
WO1993020694A1 (fr) * 1992-04-14 1993-10-28 Signal Pharmaceuticals, Inc. Procede et composition pour l'inhibition de la replication virale
US5723594A (en) * 1995-06-07 1998-03-03 Nexstar Pharmaceuticals, Inc. High affinity PDGF nucleic acid ligands

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1620450A2 (fr) * 2003-04-13 2006-02-01 Enzon Pharmaceuticals, Inc. Promedicaments oligonucleotidiques polymeres
EP1620450A4 (fr) * 2003-04-13 2011-01-19 Enzon Pharmaceuticals Inc Promedicaments oligonucleotidiques polymeres
US8168605B2 (en) 2003-04-13 2012-05-01 Enzon Pharmaceuticals, Inc. Polymeric oligonucleotide prodrugs
US9506897B2 (en) 2012-11-16 2016-11-29 Agilent Technologies, Inc. Methods and compositions for improved ion-exchange chromatography
CN108676130A (zh) * 2018-06-06 2018-10-19 河南师范大学 用于药物载体的还原响应型核交联聚合物MPEG-NSASS-Chol胶束的制备
CN108676130B (zh) * 2018-06-06 2020-08-14 河南师范大学 用于药物载体的还原响应型核交联聚合物MPEG-NSASS-Chol胶束的制备

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