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WO2016030863A1 - Composés et méthodes de traitement des infections virales - Google Patents

Composés et méthodes de traitement des infections virales Download PDF

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Publication number
WO2016030863A1
WO2016030863A1 PCT/IB2015/056539 IB2015056539W WO2016030863A1 WO 2016030863 A1 WO2016030863 A1 WO 2016030863A1 IB 2015056539 W IB2015056539 W IB 2015056539W WO 2016030863 A1 WO2016030863 A1 WO 2016030863A1
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Prior art keywords
oligonucleotide
hepatitis
independently
pharmaceutically acceptable
infection
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Inventor
Zhi Hong
Robert K. Hamatake
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GlaxoSmithKline Intellectual Property Development Ltd
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GlaxoSmithKline Intellectual Property Development Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19—Cytokines; Lymphokines; Interferons
    • A61K38/21—Interferons [IFN]
    • A61K38/212—IFN-alpha
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/7105—Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19—Cytokines; Lymphokines; Interferons
    • A61K38/21—Interferons [IFN]
    • A61K38/215—IFN-beta
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19—Cytokines; Lymphokines; Interferons
    • A61K38/21—Interferons [IFN]
    • A61K38/217—IFN-gamma
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/2292—Thymosin; Related peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/549—Sugars, nucleosides, nucleotides or nucleic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A61K47/6425—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent the peptide or protein in the drug conjugate being a receptor, e.g. CD4, a cell surface antigen, i.e. not a peptide ligand targeting the antigen, or a cell surface determinant, i.e. a part of the surface of a cell

Definitions

  • HBV is a small enveloped DNA virus belonging to the hepadnavirus family.
  • the virus replicates through an RNA intermediate form by reverse transcription, which in practice relates them to retroviruses, like HIV. Although replication takes place in the liver, the virus spreads to the blood where viral proteins and antibodies against them l are found in infected people. HBV is many times more infectious than HIV due to the greater concentrations of HBV virus found in the bloodstream at any given time.
  • nucleic acid polymers are broad- spectrum antiviral compounds that use the sequence-independent properties of NAP.
  • oligonucleotide compositions and therapeutic agents have been described and can be made as disclosed in U.S. Patents, e.g. US8,598,139, US8,507,455, US8,450,467, US8,344, 125 and US8, 106,022 and US Patent Publications e.g. US20130338210 and US20140099666;. the contents of which are hereby incorporated by reference herein in their entirety.
  • the present invention provides a method for the treatment of a hepatitis B infection or hepatitis B/hepatitis D co-infection, the method comprising administering to a subject in need of such treatment a sequence independent oligonucleotide conjugated with one or more GalNAc carbohydrate residues.
  • olinonucleotide corresponding to any one of SEQ ID NOs: 1-10 conjugated with one or more GalNAc carbohydrate residues.
  • the administration of the sequence independent oligonucleotide conjugated with one or more GalNAc carbohydrate residues inhibits the release of hepatitis B surface antigen (HBsAg) from infected hepatocytes.
  • the administration of the sequence independent oligonucleotide conjugated with one or more GalNAc carbohydrate residues comprises the administration of the compound depicted in Scheme 1 .
  • heteroaryl refers to an aromatic 5-8 membered monocyclic, 8-12 membered bicyclic, or 1 1 -14 membered tricyclic ring system having 1 -3 heteroatoms if monocyclic, 1 -6 heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms selected from O, N, or S (e.g., carbon atoms and 1 -3, 1-6, or 1 -9 heteroatoms of N, O, or S if monocyclic, bicyclic, or tricyclic, respectively), wherein 0, 1 , 2, 3, or 4 atoms of each ring may be substituted by a substituent.
  • oxo refers to an oxygen atom, which forms a carbonyl when attached to carbon, an N-oxide when attached to nitrogen, and a sulfoxide or sulfone when attached to sulfur.
  • substituted refers to the replacement of one or more hydrogen radicals in a given structure with the radical of a specified substituent including, but not limited to: halo, alkyl, alkenyl, alkynyl, aryl, heterocyclyl, thiol, alkylthio, arylthio,
  • HBsAg may also function to directly block activation of adaptive and innate immune responses to HBV infection the presence of this functionality in human HBV infection and HBV/HDV co-infection and its impact on the activity of immunotherapeutic agents has not been investigated or
  • HBeAg and HBcAg have also been shown to have immunoinhibitory properties.
  • a treatment against HBV infection and HBV/HDV co- infection which consists of a first pharmaceutically acceptable oligonucleotide conjugate agent capable of removing HBsAg from the blood and an immunotherapeutic agent which stimulates immune function.
  • a combination treatment allows circulating anti-HBsAg antibodies to directly attack the circulating virus and virus producing cells and, in the absence of the immuno-inhibitory properties of HBsAg, is exected to lead to an
  • phosphorodithioate linkages and/or methylphosphonate linkages. While modified linkages are useful, the ONs can include phosphodiester linkages. Additional useful modifications include, without restriction, modifications at the 2'-position of the sugar including 2'-0-alkyl modifications such as 2'-0-methyl modifications, 2' O-methoxyethyl (2' MOE), 2'-amino modifications, 2'-halo modifications such as 2'-fluoro; acyclic nucleotide analogs. Other 2' modifications are also known in the art and can be used such as locked nucleic acids.
  • the ON has modified linkages throughout or has every linkage modified, e.g., phosphorothioate; has a 3'- and/or 5'-cap; includes a terminal 3'-5' linkage; the ON is or includes a concatemer consisting of two or more ON sequences joined by a linker(s).
  • Base modifications can include 5'methylation of the cytosine base (5' methylcytosine or in the context of a nucleotide, 5' methylcytidine) and/or 4'thioation of the uracil base (4'thiouracil or in the context of a nucleotide, 4'thiouridine).
  • a phosphorothioated oligonucleotide from 20-120 nucleotides in length comprising repeats of the sequence CA;
  • a phosphorothioated oligonucleotide from 20-120 nucleotides in length comprising repeats of the sequence GT;
  • a phosphorothioated oligonucleotide from 20-120 nucleotides in length comprising repeats of the sequence UG;
  • the antisense strand (or guide strand) is incorporated into the RISC(RNA-induced silencing complex) which uses guide-strand targeted hybridization with the complementary region on a target mRNA to effect its cleavage by the catylytic component of the RISC called Argonaute.
  • RISC RNA-induced silencing complex
  • antiviral ON refers to any antisense ON, siRNA, shRNA or NAP or any ON conjugate, siRNA conjute, shRNA conjugate or NAP conjugate, which by virtue of its specific biochemical activity (whether sequence dependent or sequence independent) has the ability to directly or indirectly inhibit some aspect of viral replication or to directly or indirectly enhance the host's ability to clear the viral infection by immunological or other mechanisms.
  • ON chelate complex or “ONC chelate complex” refers to a complex of two or more ONs or two or more ONCs in solution linked intermolecularly by a divalent metal cation as described in International application publication no. WO 2012/021985 and U.S. application publication no. 2012/0046348, which are incorporated herein by reference in their entirety.
  • ON or ONC chelate complexes can be formed with antisense ONs or ON conjugatess, siRNAs or siRNA conjugates, or NAPs or NAP conjugates.
  • Examples of ONs which can be useful for preparation of oligonucleotide conjugates in the current disclosure include oligonucleotides comprising SEQ ID NOs 1 -10.
  • interferon-based therapy Another important limitation of interferon-based therapy is that it elicits only a moderate level ( ⁇ 50 mlU/ml) of anti-HBs production in a very small proportion of patients on treatment (Reijnders et al., 201 1 , J. Hepatol., 54: 449-454; Harayiannis et al., 1990, J. Hepatol., 10: 350-352) after 48 weeks of exposure.
  • These important limitations are likely critical factors underlying the achievement of a sustained virologic response only in a limited number of patients after immunotherapy.
  • Pegylation is also known to increase the water solubility of the polypeptide in question, improving its ease of formulation.
  • pegylated polypeptides include: Mircera.TM. a pegylated form of erythropoietin; NeulastaTM, a pegylated form of human granulocyte colony-stimulating factor; PegasysTM a pegylated form of human interferon a-2a; Peg- IntronTM, a pegylated form of human interferon a-2b; and pegylated interferon ⁇ 1 (which is currently in clinical development).
  • immunotherapeutic agents which have not been previously shown to have useful immunotherapeutic activity in the presence of HBV proteins (e.g. in infected patients, chimpanzees or cellular models) may now be shown to have useful immunotherapeutic activity with the removal of HBsAg from the blood and may be further useful in the treatment of HBV in combination with any agent which removes HBsAg from the blood.
  • interferon v-1 b Interferon ⁇ 1 or A2 or A3;
  • Toll-like receptor agonist such as GS-9620, and ANA-773
  • any hormone shown to have antiviral activity or immunostimulatory activity such as DHEA or its metabolites.
  • any amount of HBsAg removal may provide a synergistic improvement in the activity of an immunotherapy and a fractional dose of a particular immunotherapeutic agent may result in comparable or even superior
  • a and B are each independently for each occurrence O, N(R ), S or absent;
  • alkynylheteroarylalkyl alkynylheteroarylalkenyl, alkynylheteroarylalkynyl,
  • the Linker is -[(P-Q"-R) q -X-(P'-Q"'-R') q .] q .-T-, wherein:
  • q, q' and q" are each independently for each occurrence 0-20 and wherein the repeating unit can be the same or different;
  • a cleavable linking group is one which is sufficiently stable outside the cell, but which upon entry into a target cell is cleaved to release the two parts the linker is holding together.
  • the cleavable linking group is cleaved at least 10 times or more, preferably at least 100 times faster in the target cell or under a first reference condition (which can, e.g., be selected to mimic or represent intracellular conditions) than in the blood of a subject, or under a second reference condition (which can, e.g., be selected to mimic or represent conditions found in the blood or serum).
  • a cleavable linkage group such as a disulfide bond can be susceptible to pH.
  • the pH of human serum is 7.4, while the average intracellular pH is slightly lower, ranging from about 7.1 -7.3.
  • Endosomes have a more acidic pH, in the range of 5.5-6.0, and lysosomes have an even more acidic pH at around 5.0.
  • Some linkers will have a cleavable linking group that is cleaved at a preferred pH, thereby releasing the cationic lipid from the ligand inside the cell, or into the desired compartment of the cell.
  • Preferred embodiments are » 0 » P(0)(OH) » 0 » , ⁇ 0 ⁇ P(S)(OH) ⁇ 0 ⁇ , ⁇ 0 ⁇ P(S)(SH) ⁇ 0 ⁇ , ⁇ S ⁇ P(0)(OH) ⁇ 0 ⁇ , ⁇ 0 ⁇ P(0)(OH) ⁇ S ⁇ , ⁇ S ⁇ P(0)(OH) ⁇ S-, ⁇ 0 ⁇ P(S)(OH) ⁇ S ⁇ , ⁇ S ⁇ P(S)(OH) ⁇ 0 ⁇ , ⁇ 0 ⁇ P(0)(H) ⁇ 0 ⁇ , ⁇ 0 ⁇ P(S)(H) ⁇ 0 ⁇ , -S- P(0)(H) ⁇ 0 ⁇ , ⁇ S ⁇ P(S)(H) ⁇ 0 ⁇ , ⁇ S ⁇ P(0)(H) ⁇ S ⁇ , ⁇ 0 ⁇ P(S)(H) ⁇ S ⁇ .
  • a preferred embodiments are » 0 » P(0)(OH) » 0 » , ⁇ 0 ⁇ P(S)(OH) ⁇ 0 ⁇ , ⁇ 0 ⁇ P(S)(SH) ⁇ 0 ⁇
  • Ester-based cleavable linking groups are cleaved by enzymes such as esterases and amidases in cells.
  • ester-based cleavable linking groups include but are not limited to esters of alkylene, alkenylene and alkynylene groups.
  • Ester cleavable linking groups have the general formula ⁇ C(0)0 ⁇ , or -OC(O)-. These candidates can be evaluated using methods analogous to those described above.
  • the peptide based cleavage group is generally limited to the peptide bond (i.e., the amide bond) formed between amino acids yielding peptides and proteins and does not include the entire amide functional group.
  • Peptide-based cleavable linking groups have the general formula— NHCHR A C(0)NHCHR B C(0)—, where R A and R B are the R groups of the two adjacent amino acids. These candidates can be evaluated using methods analogous to those described above.
  • the oligonucleotide conjugate compound has at least one subunit of formula (1 ): Formula (I) wherein:
  • a and B are each independently for each occurrence O, N(R ), S or absent;
  • R is
  • R is
  • monomer of formula (I) has the structure
  • R is
  • formula (I) has the structure
  • formula (I) has the structure
  • R is O or S.
  • the invention features, an oligonucleotide conjugate agent comprising at least one monomer of formula (I).
  • the oligonucleotide conjugate agent will comprise 1 , 2, 3, 4 or 5 monomers of formula (I), more preferably 1 , 2 or 3 monomers of formula (I), more preferably 1 or 2 monomers of formula (I), even more preferably only one monomer of formula (I).
  • all the monomers of formula (I) are on the same strand of the oligonucleotide conjugate agent.
  • all monomers of formula (I) in an oligonucleotide conjugate agent are the same.
  • the monomers of formula (I) in an oligonucleotide conjugate agent are all different.
  • the monomers of formula (I) will be next to each other in the oligonucleotide conjugate agent.
  • the monomer of formula (I) will be on the 5'- end, 3'-end, at an internal position, both the 3'- and the 5'-end, both 5'-end and an internal position, both 3'-end and internal position, and at all three positions (5'-end, 3'-end and an internal position) of the oligonucleotide conjugate agent.
  • the oligonucleotide conjugate agent described herein does not contain homology to the target gene, but rather comprises a random or repeating AC or CA sequence and may be referred to as a randomer.
  • the oligonucleotide conjugate or randomer conjugate may be of sufficient length in terms of nucleotides, such that the oligonucleotide conjugate agent, or a fragment thereof, can modulate expression of the target gene.
  • nucleotide or ribonucleotide is sometimes used herein in reference to one or more monomeric subunits of an oligonucleotide conjugate agent. It will be understood herein that the usage of the term "ribonucleotide" or
  • formula (I) has the structure:
  • formula (I) has the structure:
  • formula (I) has the structure:
  • formula (I) has the structure:
  • R is OH or NHCOOH.
  • monomer of formula (I) is linked to the oligonucleotid
  • formula (I) has the structure
  • formula (I) has the structure
  • the oligonucleotide X or Y is connected to he at least one subunit of formula (I) at the 3'-end of the oligonucleotide.
  • Oligonucleotide conjugate agents discussed herein include unmodified oligonucleotides as well as oligonucleotides which have been modified, e.g., to improve efficacy, and polymers of nucleoside surrogates.
  • Unmodified oligonucleotide refers to a molecule in which the components of the nucleic acid, namely sugars, bases, and phosphate moieties, are the same or essentially the same as that which occur in nature, for example as occur naturally in the human body.
  • Modified oligonucleotide refers to a molecule in which one or more of the components of the nucleic acid, namely sugars, bases, and phosphate moieties, are different from that which occur in nature, for example, different from that which occurs in the human body. While they are referred to as modified
  • deoxyribophosphate or ribophosphate backbone is replaced with a non-deoxyribophosphate or nonribophosphate construct e.g., non-charged mimics of the ribophosphate or deoxyribophsophate backbone. Examples of all of the above are discussed herein.
  • nucleic acids are polymers of subunits, many of the modifications described below occur at a position which is repeated within a nucleic acid, e.g., a modification of a base, or a phosphate moiety, or the a non-linking O of a phosphate moiety.
  • the modification will occur at all of the subject positions in the nucleic acid but in many cases it will not.
  • a modification may only occur at a 3' or 5' terminal position, may only occur in a terminal regions, e.g., at a position on a terminal nucleotide or in the last 2, 3, 4, 5, or 10 nucleotides of a strand.
  • a modification may occur, e.g., a phosphorothioate modification, at a non-linking O position and may only occur at one or both termini, may only occur in a terminal regions, e.g., at a position on a terminal nucleotide or may occure in the last 2, 3, 4, 5, or 10 nucleotides of a strand, or may occur in all positions of the oligonucleotide.
  • the 5' end or ends can be phosphorylated.
  • cytidine will be modified, e.g., with a modification described herein.
  • Modifications can include, e.g., 5-methyl cytidine, and may include the use of modifications at the 2' OH group of the ribose sugar, e.g., the use of 2'-0-methyl ribose sugars.
  • Other modifications may include deoxythymidine, modifications in the phosphate group, e.g., phosphorothioate modifications.
  • nucleotide scaffole of the nucleic acid polymer has the structure of Formula (B):
  • R is H or OH or OR'
  • the scaffold presented above in Formula B represents a portion of an oligonucleic acid.
  • the basic components are the ribose or deoxyribose sugar, the base, the terminal phosphates, and phosphate internucleotide linkers.
  • the bases are naturally occurring bases, e.g., adenine, uracil, guanine or cytosine
  • the sugars are the unmodified 2' hydroxyl ribose sugar or 2'-H
  • W, X, Y, and Z are all O
  • Formula B represents a naturally occurring unmodified oligonucleotide.
  • Modified nucleic acids and nucleotide surrogates can include one or more of: (i) alteration, e.g., replacement, of one or both of the non-linking (X and Y) phosphate oxygens and/or of one or more of the linking (W and Z) phosphate oxygens (When the phosphate is in the terminal position, one of the positions W or Z will not link the phosphate to an additional element in a naturally occurring nucleic acid.
  • the W position at the 5' end of a nucleic acid and the terminal Z position at the 3' end of a nucleic acid are within the term "linking phosphate oxygens" as used herein); (ii) alteration, e.g., replacement, of a constituent of the ribose or deoxyribose sugar, e.g., of the 2' hydroxyl on the ribose sugar; (iii) wholesale replacement of the phosphate moiety (bracket I) with "dephospho" linkers; (iv) modification or replacement of a naturally occurring base; (v) replacement or modification of the sugar-phosphate backbone (bracket II); (vi) modification of the 3' end or 5' end of the oligonucleotide, e.g., removal, modification or replacement of a terminal phosphate group or conjugation of a moiety, e.g., a fluorescently labeled moiety, to either the 3'
  • polyethyleneglycols PEG
  • LNA locked nucleic acids
  • AMINE NH 2 ; alkylamino, dialkylamino, heterocyclyl, arylamino, diaryl amino, heteroaryl amino, or diheteroaryl amino, ethylene diamine, polyamino
  • AMINE NH 2 ; alkylamino, dialkylamino, heterocyclyl, arylamino, diaryl amino, heteroaryl amino, or diheteroaryl amino, ethylene diamine, polyamino
  • AMINE NH 2 ; alkylamino, dialkylamino, heterocyclyl, arylamino, diaryl amino, heteroaryl amino, or diheteroaryl amino, ethylene diamine, polyamino
  • MOE methoxyethyl group
  • the sugar group can also contain one or more carbons that possess the opposite stereochemical configuration than that of the corresponding carbon in ribose.
  • a modified oligonucleotide can include nucleotides containing e.g., arabinose, as the sugar.
  • Modified oligonucleotides can also include "abasic" sugars, which lack a nucleobase at C-1 '. These abasic sugars can also be further contain modifications at one or more of the constituent sugar atoms.
  • the 2' modifications can be used in combination with one or more phosphate linker modifications (e.g., phosphorothioate).
  • phosphate linker modifications e.g., phosphorothioate
  • chimeric oligonucleotides are those that contain two or more different modifications.
  • the phosphate group can be replaced by non-phosphorus containing connectors (cf. Bracket I in Formula B above). While not wishing to be bound by theory, it is believed that since the charged phosphodiester group is the reaction center in nucleolytic degradation, its replacement with neutral structural mimics should impart enhanced nuclease stability. Again, while not wishing to be bound by theory, it can be desirable, in some embodiment, to introduce alterations in which the charged phosphate group is replaced by a neutral moiety.
  • moieties which can replace the phosphate group include siloxane, carbonate, carboxymethyl, carbamate, amide, thioether, ethylene oxide linker, sulfonate, sulfonamide, thioformacetal, formacetal, oxime, methyleneimino,
  • replacements may include the methylenecarbonylamino and methylenemethylimino groups.
  • Oligonucleotide-mimicking scaffolds can also be constructed wherein the phosphate linker and ribose sugar are replaced by nuclease resistant nucleoside or nucleotide surrogates (see Bracket II of Formula B above). While not wishing to be bound by theory, it is believed that the absence of a repetitively charged backbone diminishes binding to proteins that recognize polyanions (e.g., nucleases). Again, while not wishing to be bound by theory, it can be desirable in some embodiment, to introduce alterations in which the bases are tethered by a neutral surrogate backbone.
  • Examples include the mophilino, cyclobutyl, pyrrolidine and peptide nucleic acid (PNA) nucleoside surrogates.
  • PNA peptide nucleic acid
  • terminal modifications include dyes, intercalating agents (e.g., acridines), cross-linkers (e.g., psoralene, mitomycin C), porphyrins (TPPC4, texaphyrin, Sapphyrin), polycyclic aromatic hydrocarbons (e.g., phenazine, dihydrophenazine), artificial endonucleases (e.g., EDTA), lipophilic carriers (e.g., cholesterol, cholic acid, adamantane acetic acid, 1-pyrene butyric acid, dihydrotestosterone, 1 ,3-Bis-O(hexadecyl)glycerol, geranyloxyhexyl group,
  • absorption facilitators e.g., aspirin, vitamin E, folic acid
  • synthetic ribonucleases e.g., imidazole, bisimidazole, histamine, imidazole clusters, acridine-imidazole conjugates, Eu 3+ complexes of tetraazamacrocycles.
  • Terminal modifications can be added for a number of reasons, including as discussed elsewhere herein to modulate activity or to modulate resistance to degradation.
  • Terminal modifications useful for modulating activity include modification of the 5' end with phosphate or phosphate analogs.
  • oligonucleotide conjugate agents are 5' phosphorylated or include a phosphoryl analog at the 5' prime terminus.
  • Suitable modifications include: 5'-monophosphate ((HO) 2 (0)P-0-5'); 5'- diphosphate ((HO) 2 (0)P-0-P(HO)(0)-0-5'); 5'-triphosphate ((HO) 2 (0)P-0-(HO)(0)P-0- P(HO)(0)-0-5'); 5'-guanosine cap (7-methylated or non-methylated) (7m-G-0-5'-(HO)(0)P- -0-(HO)(0)P-0-P(HO)(0)-0-5'); 5'-adenosine cap (Appp), and any modified or unmodified nucleotide cap structure (N-0-5'-(HO)(0)P-0-(HO)(0)P-0-P(HO)(0)-0-5'); 5'-monothiophosphate (phosphorothioate; (HO) 2 (S)P-0-5'); 5'-monodithiophosphate (phosphorodithioate; (HO)(HS)(S)P
  • Terminal modifications can also be useful for increasing resistance to degradation.
  • Terminal modifications can also be useful for monitoring distribution, and in such cases the groups to be added may include fluorophores, e.g., fluorscein or an Alexa dye, e.g., Alexa 488. Terminal modifications can also be useful for enhancing uptake, useful modifications for this include cholesterol. Terminal modifications can also be useful for cross-linking an RNA agent to another moiety; modifications useful for this include mitomycin C.
  • Adenine, guanine, cytosine and uracil are the most common bases found in RNA. These bases can be modified or replaced to provide RNAs having improved properties.
  • nuclease resistant oligoribonucleotides can be prepared with these bases or with synthetic and natural nucleobases (e.g., inosine, thymine, xanthine, hypoxanthine, nubularine, isoguanisine, or tubercidine) and any one of the above modifications.
  • substituted or modified analogs of any of the above bases and "universal bases” can be employed.
  • Examples include 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 5-halouracil, 5-(2-aminopropyl)uracil, 5- amino allyl uracil, 8-halo, amino, thiol, thioalkyl, hydroxyl and other 8-substituted adenines and guanines, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7- methylguanine, 5-substituted pyrimidines
  • base changes are not used for promoting stability, but they can be useful for other reasons, e.g., some, e.g., 2,6-diaminopurine and 2 amino purine, are fluorescent. Modified bases can reduce target specificity. This may be taken into
  • carbohydrate refers to a compound which is either a carbohydrate per se made up of one or more monosaccharide units having at least 6 carbon atoms (which may be linear, branched or cyclic) with an oxygen, nitrogen or sulfur atom bonded to each carbon atom; or a compound having as a part thereof a carbohydrate moiety made up of one or more monosaccharide units each having at least six carbon atoms (which may be linear, branched or cyclic), with an oxygen, nitrogen or sulfur atom bonded to each carbon atom.
  • Representative carbohydrates include the sugars (mono-, di-, tri- and oligosaccharides containing from about 4-9 monosaccharide units), and
  • polysaccharides such as starches, glycogen, cellulose and polysaccharide gums.
  • Specific monosaccharides include C 5 and above (preferably C 5 -C 8 ) sugars; di- and trisaccharides include sugars having two or three monosaccharide units (preferably C 5 -C 8 ).
  • the term "monosaccharide” embraces radicals of allose, altrose, arabinose, cladinose, erythrose, erythrulose, fructose, D-fucitol, L-fucitol, fucosamine, fucose, fuculose, galactosamine, D-galactosaminitol, N-acetyl-galactosamine, galactose, glucosamine, N-acetyl-glucosamine, glucosaminitol, glucose, glucose-6-phosphate, gulose glyceraldehyde, L-glycero-D-mannos-heptose, glycerol, glycerone, gulose, idose, lyxose, mannosamine, mannose, mannose-6-phosphate, psicose, quinovose, quinovosamine, rhamnitol, rhamnos
  • the monosaccharide can be in D- or L configuration.
  • the terms "disaccharide”, “trisaccharide” and “polysaccharide” embrace radicals of abequose, acrabose, anucetose, amylopectin, amylose, apiose, arcanose, ascarylose, ascorbic acid, boivinose, cellobiose, cellobiose, cellulose, chacotriose, chalcose, chitin, colitose, cyclodextrin, cymarose, dextrin, 2-deoxyribose, 2deoxyglucose, diginose, digitalose, digitoxose, evalose, evemitrose, fructooligosachharide, galto-oligosaccharide, gentianose, gentiobiose, glucan, glucogen, glycogen, hamamelose, heparin, inulin, isole
  • Disaccharide also includes amino sugars and their derivatives, particularly, a mycaminose, derivatized at the C-4' position or a 4 deoxy-3- amino-glucose derivatized at the C-6' position.
  • the invention provides a sequence independent pharmaceutically acceptable oligonucleotide that is conjugated with at least one
  • carbohydrate ligand e.g., monosaccharide, disaccharide, trisaccharide, tetrasaccharide, oligosaccharide, polysaccharide.
  • these carbohydrate-conjugated sequence independent oligonucelotide target, in particular, the parenchymal cells of the liver.
  • the carbohydrate-conjugated sequence independent oligonucelotide includes more than one carbohydrate ligand, preferably two or three.
  • the carbohydrate-conjugated sequence independent oligonucelotide comprises one or more galactose moiety.
  • the carbohydrate- conjugated sequence independent oligonucelotide includes at least one (e.g., two or three or more) lactose molecules (lactose is a glucose coupled to a galactose).
  • the carbohydrate-conjugated sequence independent oligonucelotide includes at least one (e.g., two or three or more) N-Acetyl-Galactosamine (GalNAc) or N-Ac- Glucosamine (GluNAc).
  • the carbohydrate-conjugated sequence independent oligonucelotide includes at least one (e.g., two or three or more) N-Acetyl- Galactosamine (GalNAc).
  • the carbohydrate-conjugated sequence independent oligonucelotide includes at least three N-Acetyl-Galactosamine (GalNAc).
  • Carbohydrate and lipid conjugates of oligonucleotide compositions and therapeutic agents are described and can be made as disclosed in U.S. Patents, e.g. US8,598,139, US8,507,455, US8,450,467, US8,344, 125 and US8, 106,022 and US Patent Publications e.g. US20130338210 and US20140099666,. the contents of which are hereby incorporated by reference herein in their entirety.
  • the presently claimed invention involves coupling certain conjugates to the aforementioned sequence independent oligonucleotides for delivery, particularly adding carbohydrate conjugates, and more particularly coupling GalNAc conjugates, to the non-specific oligonucleotides disclosed in the above-cited patents and applications.
  • carbohydrate conjugates can be coupled via linkers as described in the cited US patents and patent publications above.
  • the resulting non-sequence specific oligonucleotide antiviral agent conjugates are envisioned to achieve greater efficacy, to exhibit longer half-life inside a patient after administration, and to improve the therapeutic index for the non-sequence specific oligonucleotide antiviral agent.
  • the invention provides an oligonucleotide conjugate agent that is conjugated with at least one carbohydrate ligand, e.g., a monosaccharide, a disaccharide, a trisaccharide, a tetrasaccharide, a oligosaccharide, a polysaccharide.
  • carbohydrate ligand e.g., a monosaccharide, a disaccharide, a trisaccharide, a tetrasaccharide, a oligosaccharide, a polysaccharide.
  • the invention features an oligonucleotide conjugate agent comprising a carbohydrate ligand, and the presence of the carbohydrate ligand can increase delivery of the oligonucleotide conjugate agent to the liver.
  • an oligonucleotide conjugate agent comprising a carbohydrate ligand can be useful for targeting a gene for which expression is undesired in the liver.
  • a human who is a candidate for treatment with a carbohydrate-conjugated oligonucleotide conjugate agent e.g., an oligonucleotide conjugate agent that targets a gene of HCV
  • can present symptoms indicative of HCV infection such as jaundice, abdominal pain, liver enlargement and fatigue.
  • a carbohydrate-conjugated oligonucleotide conjugate agent targets the 5' core region of HCV. This region lies just downstream of the ribosomal toe-print straddling the initiator methionine.
  • an oligonucleotide conjugate agent targets any one of the nonstructural proteins of HCV, such as NS3, NS4A, NS4B, NS5A, or NS5B.
  • an oligonucleotide conjugate agent targets the E1 , E2, or C gene of HCV.
  • Carbohydrate-conjugated oligonucleotide conjugate agents can also be used to treat other liver disorders, including disorders characterized by unwanted cell proliferation, hematological disorders, metabolic disorders, and disorders characterized by inflammation.
  • a proliferation disorder of the liver can be, for example, a benign or malignant disorder, e.g., a cancer, e.g., a hepatocellular carcinoma (HCC), hepatic metastasis, or hepatoblastoma.
  • a hepatic hematology or inflammation disorder can be a disorder involving clotting factors, a complement-mediated inflammation or a fibrosis, for example.
  • Metabolic diseases of the liver include dyslipidemias and irregularities in glucose regulation.
  • a liver disorder is treated by administering one or more oligonucleotide conjugate agents that have a sequence that is substantially identical to a sequence in a gene involved in the liver disorder.
  • a carbohydrate-conjugated oligonucleotide conjugate agent targets a nucleic acid expressed in the liver, such as an ApoB RNA, c-jun RNA, beta-catenin RNA, or glucose-6-phosphatase mRNA.
  • An oligonucleotide conjugate that targets glucose-6-phosphatase can be administered to a subject to inhibit hepatic glucose production, e.g., for the treatment of glucose-metabolism-related disorders, such as diabetes, e.g., type-2-diabetes mellitus.
  • the oligonucleotide conjugate agent can be administered to an individual at risk for the disorder to delay onset of the disorder or a symptom of the disorder.
  • the present invention provides a method for the treatment of a hepatitis B infection or hepatitis B/hepatitis D co-infection, the method comprising administering to a subject in need of such treatment a sequence independent oligonucleotide conjugated with one or more GalNAc carbohydrate residues.
  • the present invention provides a method for the treatment of a hepatitis B infection or hepatitis B/hepatitis D co-infection, the method comprising administering to a subject in need of such treatment a first pharmaceutically acceptable agent comprising a sequence independent oligonucleotide conjugated with one or more GalNAc carbohydrate residues in combination with a second pharmaceutically acceptable agent that stimulates immune function.
  • the present invention provides a method for the treatment of a hepatitis B infection or hepatitis B/hepatitis D co-infection, the method comprising administering to a subject in need of such treatment a first pharmaceutically acceptable agent comprising a sequence independent oligonucleotide conjugated with one or more GalNAc carbohydrate residues in combination with a second pharmaceutically acceptable agent that stimulates immune function and a third pharmaceutically acceptable agent comprising an antiviral compound.
  • the administration of the sequence independent oligonucleotide conjugated with one or more GalNAc carbohydrate residues inhibits the release of hepatitis B surface antigen (HBsAg) from infected hepatocytes.
  • HBsAg hepatitis B surface antigen
  • oligonucleotide conjugate is an oligonucleotide- carbohydrate conjugate
  • oligonucleotide conjugate is an oligonucleotide-carbohydrate conjugate.
  • a method for the treatment of HBV infection or HBV/HDV co-infection in a subject requiring such treatment comprising the administration of an effective dosing regimen of a first pharmaceutically acceptable agent that is an oligonucleotide conjucate which inhibits the release of HBV subviral particles from infected cells, and an effective dosing regimen of a second pharmaceutically acceptable immunotherapeutic agent which stimulates immune function, wherein the oligonucleotide conjugate is an oligonucleotide-carbohydrate conjugate.
  • a method for the treatment of HBV infection or HBV/HDV co-infection in a subject requiring such treatment comprising the administration of a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which inhibits the formation of HBV subviral particles in infected cells, and a second pharmaceutically acceptable immunotherapeutic agent which stimulates immune function, wherein the oligonucleotide conjugate is an oligonucleotide-carbohydrate conjugate.
  • a method for the treatment of HBV infection or HBV/HDV co-infection in a subject requiring such treatment comprising the administration of the first pharmaceutically acceptable agent that is an oligonucleotide conjudate and second pharmaceutically acceptable agent as described above, in combination in a single pharmaceutical composition or in two different pharmaceutical compositions given by the same route of administration, wherein the oligonucleotide conjugate is an oligonucleotide-carbohydrate conjugate.
  • a method for the treatment of HBV infection or HBV/HDV co-infection in a subject requiring such treatment comprising the administration of the first pharmaceutically acceptable agent that is an oligonucleotide conjugate and second pharmaceutical agent as described above, simultaneously in a patient, whether given by the same or different routes of administration.
  • a first pharmaceutically acceptable agent that is an oligonucleotide which removes hepatitis B surface antigen from the blood in combination with a second pharmaceutically acceptable immunotherapeutic agent that stimulates immune function for the treatment of hepatitis B infection or hepatitis B/hepatitis D co-infection, wherein the oligonucleotide conjugate is an oligonucleotide-carbohydrate conjugate.
  • a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which inhibits the release of HBsAg from infected cells
  • a second pharmaceutically acceptable immunotherapeutic agent which stimulates immune function for the treatment of hepatitis B infection or hepatitis B/hepatitis D co- infection
  • the oligonucleotide conjugate is an oligonucleotide-carbohydrate conjugate.
  • a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which inhibits the formation of HBV subviral particles in infected cells and a second pharmaceutically acceptable immunotherapeutic agent which stimulates immune function for the treatment of hepatitis B infection or hepatitis B/hepatitis D co-infection, wherein the oligonucleotide conjugate is an oligonucleotide- carbohydrate conjugate.
  • a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which removes hepatitis B surface antigen from the blood in combination with a second pharmaceutically acceptable immunotherapeutic agent that stimulates immune function in the manufacture of a medicament for the treatment of hepatitis B infection or hepatitis B/hepatitis D co-infection, wherein the oligonucleotide conjugate is an oligonucleotide-carbohydrate conjugate.
  • a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which removes hepatitis B surface antigen from the blood in combination with a second pharmaceutically acceptable immunotherapeutic agent that stimulates immune function in the manufacture of a medicament for the treatment of hepatitis B infection or hepatitis B/hepatitis D co-infection, wherein the oligonucleotide- carbohydrate conjugate has an oligonucleotide sequence of any one of SEQ ID NOs: 1 , 2, 3, 4, 5, 6, 7, 8, 9 or 10.
  • a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which removes hepatitis B surface antigen from the blood in combination with a second pharmaceutically acceptable immunotherapeutic agent that stimulates immune function in the manufacture of a medicament for the treatment of hepatitis B infection or hepatitis B/hepatitis D co-infection, wherein the oligonucleotide- carbohydrate conjugate has the structure depicted in Scheme 1 .
  • compositions for the treatment of hepatitis B infection or hepatitis B/hepatitis D co-infection comprising an effective dose of a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which removes hepatitis B surface antigen from the blood and an effective dosing regimen of a second pharmaceutically acceptable
  • oligonucleotide conjugate is an oligonucleotide-carbohydrate conjugate.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood inhibits the formation of HBV subviral particles.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood inhibits the intracellular transit of HBV subviral particles.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood inhibits the release of HBV subviral particles into the blood.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood inhibits the release of hepatitis B surface antigen from the infected cell.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood inhibits the synthesis of HBsAg and or another viral protein.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood inhibits the synthesis or function of apolipoprotein H.
  • phosphorothioated oligonucleotide from 20-120 nucleotides in length comprising repeats of the sequence AC.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood is nucleic acid polymer comprising a
  • phosphorothioated oligonucleotide from 20-120 nucleotides in length comprising the repeats of the sequence CA.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood is a nucleic acid polymer comprising a
  • phosphorothioated oligonucleotide from 20-120 nucleotides in length comprising the repeats of the sequence TG.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood is a nucleic acid polymer comprising a
  • phosphorothioated oligonucleotide from 20-120 nucleotides in length comprising the repeats of the sequence GT.
  • nucleic acid polymer of the present invention is 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
  • oligonucleotide conjugate agent further comprises at least one 2' ribose modification.
  • oligonucleotide conjugate agent further comprises all riboses having a 2' modification.
  • nucleic acid polymer of the present invention is 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
  • oligonucleotide conjugate agent further comprises at least one 2' O methyl ribose modification.
  • nucleic acid polymer of the present invention is 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
  • oligonucleotide conjugate agent further comprises all riboses having the 2' O methyl modification.
  • nucleic acid polymer of the present invention is 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
  • oligonucleotide conjugate agent further comprises at least one 5'methylcytosine.
  • nucleic acid polymer of the present invention is 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
  • oligonucleotide conjugate agent further comprises at least one 2' ribose modification and at least one 5' methylcytosine.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood comprises an oligonucleotide selected from the group consisting of SEQ ID NOs: 1-10.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood is an oligonucleotide conjugate chelate complex comprising an oligonucleotide selected from the group consisting of SEQ ID NOs: 1 -10.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood comprises an oligonucleotide consisting of SEQ ID NO: 2.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood comprises an oligonucleotide conjugate chelate complex comprising SEQ ID NO: 2.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood is an oligonucleotide conjugate chelate complex comprising an oligonucleotide conjugate comprising a sequence selected from the group consisting of SEQ ID NO: 10.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood comprises an antisense oligonucleotide conjugate which targets any portion of any HBV mRNA.
  • the oligonucleotide conjugate is further formulated as an oligonucleotide conjugate chelate complex.
  • the oligonucleotide conjugate agent removing hepatitis B surface antigen from the blood comprises an oligonucleotide aptamer or Speigelmer targeting the hepatitis B surface antigen.
  • composition comprising an
  • immunostimulatory oligonucleotide including IMO-2055 or IMO-2125;
  • a small molecule Tolllike receptor (TLR) agonist including GS-9620 or ANA-773; and Any antiviral or
  • immunostimulatory hormone including DHEA or its metabolites.
  • the immunotherapeutic agent stimulating immune function comprises one or more compounds selected from the group consisting of: A non-CpG immunostimulatory oligonucleotide including IMO-2055 or IMO-2125; A small molecule Toll-like receptor (TLR) agonist including GS-9620 or ANA-773; and any antiviral or immunostimulatory hormone including DHEA or its metabolites.
  • a non-CpG immunostimulatory oligonucleotide including IMO-2055 or IMO-2125
  • a small molecule Toll-like receptor (TLR) agonist including GS-9620 or ANA-773
  • any antiviral or immunostimulatory hormone including DHEA or its metabolites.
  • first and second pharmaceutically acceptable agents are formulated within the same pharmaceutical composition.
  • first and second agents are formulated within separate pharmaceutical compositions.
  • first and second agents are formulated for a simultaneous administration.
  • first and second agents are formulated for an administration by a different route.
  • the first and second agents are formulated for an administration using one or more of the following: oral ingestion, aerosol inhalation, subcutaneous injection, intramuscular injection, intraperitoneal injection, intravenous injection and intravenous infusion.
  • composition comprising an oligonucleotide conjugate agent removing HBsAg from the blood further comprises one or more molecules selected from the group consisting of:
  • nucleic acid polymer selected from the following:
  • oligonucleotide from 20-120 nucleotides in length
  • a phosphorothioated oligonucleotide from 20-120 nucleotides in length
  • oligonucleotide from 20-120 nucleotides in length
  • oligonucleotide from 20-120 nucleotides in length comprising repeats of the sequence GT wherein the nucleic acid polymer further comprises one or more GalNAc residues;
  • a second immunotherapeutic agent comprising one or more molecules from the group consisting of:
  • any polypeptide shown to have antiviral activity or immunostimulatory activity v. an immunostimulatory oligonucleotide including IMO-2055 or IMO-2125; w. a small molecule Toll-like receptor (TLR) agonist including GS-9620 or ANA- 773; and x. any antiviral or immunostimulatory hormone including DHEA or its metabolites.
  • TLR Toll-like receptor
  • oligonucleotide can be further formulated as an oligonucleotide conjugate chelate complex:
  • nucleic acid polymer selected from the following:
  • a phosphorothioated oligonucleotide from 20-120 nucleotides in length
  • oligonucleotide from 20-120 nucleotides in length
  • oligonucleotide from 20-120 nucleotides in length
  • oligonucleotide from 20-120 nucleotides in length
  • nucleic acid polymer of of a., b., c, and/ or d. further comprises one or more GalNAc residues; and e. the compound depicted in Scheme 1 .
  • the uses or method of treatments described above further comprise administering or using concurrently a third pharmaceutically acceptable agent selected from the following:
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises SEQ ID NO: 2 and a second pharmaceutically acceptable agent which comprises pegylated interferon a-2a.
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises an oligonucleotide chelate conjugate complex comprising SEQ ID NO: 3 and a second pharmaceutically acceptable agent which comprises pegylated interferon a-2a.
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises SEQ ID NO: 2 and a second pharmaceutically acceptable agent which comprises thymosin c .
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent which comprises an oligonucleotide chelate conjugate complex comprising SEQ ID NO: 2 and a second pharmaceutically acceptable agent which comprises thymosin a1 .
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises SEQ ID NO: 3 and a second pharmaceutically acceptable agent which comprises thymosin .alpha.1 .
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises SEQ ID NO: 10 and a second pharmaceutically acceptable agent which comprises thymosin a1 a.
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises an oligonucleotide chelate conjugate complex comprising SEQ ID NO: 10 and a second pharmaceutically acceptable agent which comprises thymosin a1 .
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises SEQ ID NO: 2 and a second pharmaceutically acceptable agent which comprises pegylated interferon a-2b.
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises an oligonucleotide chelate conjugate complex comprising SEQ ID NO: 2 and a second pharmaceutically acceptable agent which comprises pegylated interferon a-2b.
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises SEQ ID NO: 3 and a second pharmaceutically acceptable agent which comprises pegylated interferon a-2b.
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises SEQ ID NO: 10 and a second pharmaceutically acceptable agent which comprises pegylated interferon a-2b.
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises an oligonucleotide chelate conjugate complex comprising SEQ ID NO: 10 and a second pharmaceutically acceptable agent which comprises pegylated interferon a-2b.
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises SEQ ID NO: 2 and a second pharmaceutically acceptable agent which comprises pegylated interferon ⁇ 1 .
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises an oligonucleotide chelate conjugate complex comprising SEQ ID NO: 2 and a second pharmaceutically acceptable agent which comprises pegylated interferon ⁇ 1 .
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises SEQ ID NO: 3 and a second pharmaceutically acceptable agent which comprises pegylated interferon ⁇ 1 .
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises an oligonucleotide chelate conjugate complex comprising SEQ ID NO: 3 and a second pharmaceutically acceptable agent which comprises pegylated interferon ⁇ 1 .
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises SEQ ID NO: 10 and a second pharmaceutically acceptable agent which comprises pegylated interferon ⁇ 1 .
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises an oligonucleotide chelate conjugate complex comprising SEQ ID NO: 10 and a second pharmaceutically acceptable agent which comprises pegylated interferon ⁇ 1 .
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises SEQ ID NO: 2 and a second pharmaceutically acceptable agent which comprises GS-9620.
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises an oligonucleotide chelate conjugate complex comprising SEQ ID NO: 2 and a second pharmaceutically acceptable agent which comprises GS-9620.
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises an oligonucleotide chelate conjugate complex comprising SEQ ID NO: 3 and a second pharmaceutically acceptable agent which comprises GS-9620.
  • a method for the treatment of or the use of the following in the treatment of hepatitis B infection or of hepatitis B/hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises SEQ ID NO: 10 and a second pharmaceutically acceptable agent which comprises GS- 9620.
  • a first pharmaceutically acceptable agent that is an oligonucleotide conjugate which comprises an oligonucleotide chelate conjugate complex comprising SEQ ID NO: 10 and a second pharmaceutically acceptable agent which comprises GS-9620.
  • a method for the treatment of Hepatitis B infection or of Hepatitis D co-infection comprising administering to a patient in need of such treatment a first pharmaceutically acceptable agent that is an oligonucleotide conjug :
  • a and B are each independently for each occurrence O, N(R N ), S or absent;
  • R is independently for each occurrence H or C C 6 alkyl
  • X and Y are each independently for each occurrunce H, OH, a hydroxyl protecting group, a phosphate group, a phosphodiester group, an activated phosphate group, an activated phosphite group, a phosphoramidite, a solid support, - P(Z')(Z")0-nucleoside, -P(Z')(Z")0-oligonucleotide, a lipid, a PEG, a steroid, a polymer, a nucleotide, a nucleoside, -P(Z')(Z")0-Linker-OP(Z"')(Z"")0-oligonucleotide, an
  • oligonucleotide ⁇ P(Z')(Z")-formula (I), -P(Z')(Z")-, -Linker-R, or absent;
  • L G is independently for each occurrence a ligand including a carbohydrate, a monosaccharide, a disaccharide, a trisaccharide, a tetrasaccharide, a polysaccharide; and Z', Z", Z'" and Z"" are each independently for each occurrence O or S; and a second therapeutically active agent.
  • n 19-59.
  • a method for the treatment of Hepatitis B infection or of Hepatitis D co-infection as described comprising administering to a patient in need of such treatment the oligonucleotide conjugate therapeutic composition as described, further wherein only X or only Y is present in the oligonucleotide conjugate but not both, wherein X or Y is independently an oligonucleotide or absent, and wherein the oligonucleotide X or Y comprises any of SEQ ID NOs. 1 -10.
  • a method for the treatment of Hepatitis B infection or of Hepatitis D co-infection as described comprising administering to a patient in need of such treatment the oligonucleotide conjugate therapeutic composition as described, further wherein only X or only Y is present in the oligonucleotide conjugate but not both, wherein X or Y is independently an oligonucleotide or absent, and wherein the oligonucleotide X or Y has the following structure:
  • n 19-60;
  • R 2 is H or CH 3 .
  • a method for the treatment of Hepatitis B infection or of Hepatitis D co-infection as described comprising administering to a patient in need of such treatment an oligonucleotide conjugate therapeutic composition wherein the oligonucleotide conjugate has a structure as depicted in Scheme 1 .
  • a method for the treatment of Hepatitis B infection or of Hepatitis D co-infection as described comprising administering to a patient in need of such treatment the oligonucleotide conjugate therapeutic composition as described, further wherein only X or only Y is present in the oligonucleotide conjugate but not both, wherein X or Y is independently an oligonucleotide or absent, and wherein the oligonucleotide X or Y has the structure
  • oligonucleotide conjugate therapeutic composition further comprises a second therapeutically active agent.
  • an oligonucleotide conjugate comprising at least one subunit of formula (I):
  • a and B are each independently for each occurrence O, N(R N ), S or absent;
  • R N is independently for each occurrence H or C C 6 alkyl
  • X and Y are each independently for each occorrence H, OH, a hydroxyl protecting group, a phosphate group, a phosphodiester group, an activated phosphate group, an activated phosphite group, a phosphoramidite, a solid support, -P(Z')(Z")0- nucleoside, -P(Z')(Z”)0-oligonucleotide, a lipid, a PEG, a steroid, a polymer, a nucleotide, a nucleoside, -P(Z')(Z")0-Linker-OP(Z"')(Z"")0-oligonucleotide, an oligonucleotide, ⁇ P(Z')(Z")-formula (I), -P(Z')(Z")-, -Linker-R or is absent;
  • R is L G or has the structure shown below:
  • L G is independently for each occurrence a ligand including a carbohydrate, a monosaccharide, a disaccharide, a trisaccharide, a tetrasaccharide, a
  • Another embodiment provides an oligonucleotide conjugate as described herein, wherein X or Y is independently an oligonucleotide or absent and wherein only X or only Y is present in the oligonucleotide conjugate but not both, further wherein X or Y wherein the oligonucleotide X or Y comprises a sequence selected from:
  • n 19-59.
  • n 19-60;
  • R is H or CH 3 .
  • the oligonucleotide X or Y is connected to the at least one subunit of formula (I) at an internal phosphate of the oligonucleotide.
  • R is O or S.
  • the linker is -[(P-Q"-R)q-X-(P'-Q'"-R')q.] q .-T-, wherein:
  • Q" and Q'" are each independently for each occurrence absent, ⁇ (CH 2 ) n ⁇ , -
  • R a is H or an amino acid side chain
  • R 1 and R 2 are each independently for each occurrence H, CH 3 , OH, SH or N(R N ) 2 ;
  • R N is independently for each occurrence H, methyl, ethyl, propyl, isopropyl, butyl or benzyl;
  • q, q' and q" are each independently for each occurrence 0-20 and wherein the repeating unit can be the same or different;
  • n is independently for each occurrence 1-20;
  • n is independently for each occurrence 0-50.
  • the Linker comprises at least one cleavable linking group.
  • the Linker is a branched Linker.
  • composition for treatment of Hepatitis B infection or of Hepatitis D co-infection for administration to a patient in need of such treatment, wherein the composition comprises a first pharmaceutically acceptable agent that is an oligonucleotide conjugate comprising at least one subunit of formula (I):
  • a and B are each independently for each occurrence O, N(R N ), S or absent;
  • R N is independently for each occurrence H or C C 6 alkyl
  • X and Y are each independently for each occorreunce H, OH, a hydroxyll protecting group, a phosphate group, a phosphodiester group, an activated phosphate group, an activated phosphite group, a phosphoramidite, a solid support, -P(Z')(Z")0- nucleoside, -P(Z')(Z")0-oligonucleotide, a lipid, a PEG, a steroid, a polymer, a nucleotide, a nucleoside, -P(Z')(Z")0-Linker-OP(Z"')(Z"")0-oligonucleotide, an oligonucleotide, ⁇ P(Z')(Z")-formula (I), -P(Z')(Z")-, -Linker-R or is absent;
  • R is L G or has the structure shown below:
  • L G is independently for each occurrence a ligand including a carbohydrate, a monosaccharide, a disaccharide, a trisaccharide, a tetrasaccharide, a
  • Z', Z", Z'" and Z"" are each independently O or S for each occurrence.
  • the therapeutic composition as described herein wherein only X or only Y is present in the oligonucleotide conjugate but not both, wherein X or Y is independently an oligonucleotide or absent, and wherein the oligonucleotide X or Y comprises a sequence selected from:
  • n 19-59.
  • the therapeutic composition as described herein wherein only X or only Y is present in the oligonucleotide conjugate but not both, wherein X or Y is independently an oligonucleotide or absent, and wherein the oligonucleotide X or Y comprises any of SEQ ID NOs. 1 -10.
  • a therapeutic composition as described herein wherein only X or only Y is present in the oligonucleotide conjugate but not both, wherein X or Y is independently an oligonucleotide or absent, and wherein the oligonucleoti
  • R 2 is H or CH 3 .
  • the therapeutic composition as described wherein only X or only Y is present in the oligonucleotide conjugate but not both, wherein X or Y is independently an oligonucleotide or absent, and wherein the oligonucleotide X or Y has the structure
  • the therapeutic composition further comprises a second therapeutically active agent.
  • the therapeutic composition as described herein, wherein the second therapeutically active agent is selected from:
  • interferon v-1 b i. ilnterferon ⁇ 1 or A2 or A3
  • n an immunostimulatory oligonucleotide including IMO-2055 or IMO-2125; o. a small molecule Toll-like receptor (TLR) agonist including GS-9620 or ANA- 773; and
  • any antiviral or immunostimulatory hormone including DHEA or its metabolites.
  • the second therapeutic agent is selected from: ribavirin; an HBV RNA replication inhibitor; an antisense oligomer; an HBV therapeutic vaccine; an HBV prophylactic vaccine; lamivudine (3TC); entecavir; tenofovir; telbivudine (LdT); adefovir; and an HBV antibody therapy (monoclonal or polyclonal).
  • Polymers are known to affect the entry of virions in cells by, in some cases, binding or adsorbing to the virion itself. This characteristic of antiviral polymers can be useful in competing with ONCs for the binding, or adsorption to the virion, the result being an increased intracellular activity of the ONCs compared to its extracellular activity.
  • HBV polymerase inhibitor such as (but are not restricted to): tenofovir disoproxil fumarate, entecavir, telbuvidine, adefovir dipivoxil or lamivudine.
  • tenofovir disoproxil fumarate e.g., tenofovir disoproxil fumarate, entecavir, telbuvidine, adefovir dipivoxil or lamivudine.
  • Such antiviral drugs can prevent the replication of the double stranded viral genome in HBV and lower the concentration of HBV virus in the blood.
  • compositions described herein may be administered by any suitable means, for example, orally, such as in the form of tablets, capsules, granules or powders; sublingually; buccally; parenterally, such as by subcutaneous, intravenous, intramuscular or injection or infusion techniques (e.g., as sterile injectable aqueous or nonaqueous solutions or suspensions); by inhalation; topically, such as in the form of a cream or ointment; or rectally such as in the form of suppositories or enema; in dosage unit formulations containing non-toxic, pharmaceutically acceptable vehicles or diluents.
  • suitable means for example, orally, such as in the form of tablets, capsules, granules or powders; sublingually; buccally; parenterally, such as by subcutaneous, intravenous, intramuscular or injection or infusion techniques (e.g., as sterile injectable aqueous or nonaqueous solutions or suspensions); by in
  • compositions suitable for administration can be incorporated into pharmaceutical compositions suitable for administration.
  • Such compositions typically include one or more species of oligonucleotide and a pharmaceutically acceptable carrier.
  • pharmaceutically acceptable carrier is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration.
  • the use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions.
  • compositions of the present invention may be administered in a number of ways depending upon whether local or systemic treatment is desired and upon the area to be treated. Administration may be topical (including ophthalmic, vaginal, rectal, intranasal, transdermal), oral or parenteral. Parenteral administration includes intravenous drip, subcutaneous, intraperitoneal or intramuscular injection, or intrathecal or intraventricular administration.
  • the route and site of administration may be chosen to enhance targeting.
  • intramuscular injection into the muscles of interest would be a logical choice.
  • Lung cells might be targeted by administering the oligonucleotide conjugate in aerosol form.
  • the vascular endothelial cells could be targeted by coating a balloon catheter with the oligonucleotide and mechanically introducing the DNA.
  • Formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders.
  • Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.
  • Coated condoms, gloves and the like may also be useful.
  • compositions for oral administration include powders or granules, suspensions or solutions in water, syrups, elixirs or non-aqueous media, tablets, capsules, lozenges, or troches.
  • carriers that can be used include lactose, sodium citrate and salts of phosphoric acid.
  • Various disintegrants such as starch, and lubricating agents such as magnesium stearate, sodium lauryl sulfate and talc, are commonly used in tablets.
  • useful diluents are lactose and high molecular weight polyethylene glycols.
  • the nucleic acid compositions can be combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring agents can be added.
  • Formulations for parenteral administration may include sterile aqueous solutions which may also contain buffers, diluents and other suitable additives.
  • Intraventricular injection may be facilitated by an intraventricular catheter, for example, attached to a reservoir.
  • the total concentration of solutes may be controlled to render the preparation isotonic.
  • Topical administration refers to the delivery to a subject by contacting the formulation directly to a surface of the subject.
  • the most common form of topical delivery is to the skin, but a composition disclosed herein can also be directly applied to other surfaces of the body, e.g., to the eye, a mucous membrane, to surfaces of a body cavity or to an internal surface.
  • topical delivery is to the skin.
  • the term encompasses several routes of administration including, but not limited to, topical and transdermal. These modes of administration typically include penetration of the skin's permeability barrier and efficient delivery to the target tissue or stratum.
  • Topical administration can be used as a means to penetrate the epidermis and dermis and ultimately achieve systemic delivery of the composition. Topical administration can also be used as a means to selectively deliver oligonucleotides to the epidermis or dermis of a subject, or to specific strata thereof, or to an underlying tissue.
  • skin refers to the epidermis and/or dermis of an animal. Mammalian skin consists of two major, distinct layers. The outer layer of the skin is called the epidermis. The epidermis is comprised of the stratum corneum, the stratum granulosum, the stratum spinosum, and the stratum basale, with the stratum corneum being at the surface of the skin and the stratum basale being the deepest portion of the epidermis. The epidermis is between 50 .mu.m and 0.2 mm thick, depending on its location on the body.
  • Beneath the epidermis is the dermis, which is significantly thicker than the epidermis.
  • the dermis is primarily composed of collagen in the form of fibrous bundles.
  • the collagenous bundles provide support for, inter alia, blood vessels, lymph capillaries, glands, nerve endings and immunologically active cells.
  • the permeability barrier provided by the skin is such that it is largely impermeable to molecules having molecular weight greater than about 750 Da.
  • mechanisms other than normal osmosis must be used.
  • Transdermal delivery is a valuable route for the administration of lipid soluble therapeutics.
  • the dermis is more permeable than the epidermis and therefore absorption is much more rapid through abraded, burned or denuded skin.
  • Inflammation and other physiologic conditions that increase blood flow to the skin also enhance transdermal adsorption. Absorption via this route may be enhanced by the use of an oily vehicle
  • transdermal route provides a potentially effective means to deliver a composition disclosed herein for systemic and/or local therapy.
  • Therapeutic Drug Carrier Systems, 1991 , p. 163), phonophoresis or sonophoresis (use of ultrasound to enhance the absorption of various therapeutic agents across biological membranes, notably the skin and the cornea) (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991 , p. 166), and optimization of vehicle characteristics relative to dose position and retention at the site of administration (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991 , p. 168) may be useful methods for enhancing the transport of topically applied compositions across skin and mucosal sites.
  • compositions and methods provided may also be used to examine the function of various proteins and genes in vitro in cultured or preserved dermal tissues and in animals.
  • the invention can be thus applied to examine the function of any gene.
  • the methods of the invention can also be used therapeutically or prophylactically.
  • diseases such as psoriasis, lichen planus, toxic epidermal necrolysis, ertythema multiforme, basal cell carcinoma, squamous cell carcinoma, malignant melanoma, Paget's disease, Kaposi's sarcoma, pulmonary fibrosis, Lyme disease and viral, fungal and bacterial infections of the skin.
  • Pulmonary delivery compositions can be delivered by inhalation by the patient of a dispersion so that the composition, for example, oligonucleotide, within the dispersion can reach the lung where it can be readily absorbed through the alveolar region directly into blood circulation.
  • a dispersion so that the composition, for example, oligonucleotide, within the dispersion can reach the lung where it can be readily absorbed through the alveolar region directly into blood circulation.
  • Pulmonary delivery can be effective both for systemic delivery and for localized delivery to treat diseases of the lungs.
  • Pulmonary delivery can be achieved by different approaches, including the use of nebulized, aerosolized, micellular and dry powder-based formulations. Delivery can be achieved with liquid nebulizers, aerosol-based inhalers, and dry powder dispersion devices. Metered-dose devices are may be used. One of the benefits of using an atomizer or inhaler is that the potential for contamination is minimized because the devices are self contained. Dry powder dispersion devices, for example, deliver drugs that may be readily formulated as dry powders. An oligonucleotide composition may be stably stored as lyophilized or spray-dried powders by itself or in combination with suitable powder carriers.
  • the delivery of a composition for inhalation can be mediated by a dosing timing element which can include a timer, a dose counter, time measuring device, or a time indicator which when incorporated into the device enables dose tracking, compliance monitoring, and/or dose triggering to a patient during administration of the aerosol medicament.
  • a dosing timing element which can include a timer, a dose counter, time measuring device, or a time indicator which when incorporated into the device enables dose tracking, compliance monitoring, and/or dose triggering to a patient during administration of the aerosol medicament.
  • the term "powder” means a composition that consists of finely dispersed solid particles that are free flowing and capable of being readily dispersed in an inhalation device and subsequently inhaled by a subject so that the particles reach the lungs to permit penetration into the alveoli.
  • the powder is said to be "respirable.”
  • the average particle size is less than about 10 .mu.m in diameter with a relatively uniform spheroidal shape distribution. In some embodiments, the diameter is less than about 7.5 .mu.m and in some embodiments less than about 5.0 .mu.m.
  • the particle size distribution is between about 0.1 .mu.m and about 5 .mu.m in diameter, sometimes about 0.3 .mu.m to about 5 .mu.m.
  • dry means that the composition has a moisture content below about 10% by weight (% w) water, usually below about 5% w and in some cases less it than about 3% w.
  • a dry composition can be such that the particles are readily dispersible in an inhalation device to form an aerosol.
  • physiologically effective amount is that amount delivered to a subject to give the desired palliative or curative effect.
  • pharmaceutically acceptable carrier means that the carrier can be taken into the lungs with no significant adverse toxicological effects on the lungs.
  • the types of pharmaceutical excipients that are useful as carrier include stabilizers such as human serum albumin (HSA), bulking agents such as carbohydrates, amino acids and polypeptides; pH adjusters or buffers; salts such as sodium chloride; and the like. These carriers may be in a crystalline or amorphous form or may be a mixture of the two.
  • HSA human serum albumin
  • bulking agents such as carbohydrates, amino acids and polypeptides
  • pH adjusters or buffers such as sodium chloride
  • salts such as sodium chloride
  • Bulking agents that are particularly valuable include compatible carbohydrates, polypeptides, amino acids or combinations thereof.
  • Suitable carbohydrates include monosaccharides such as galactose, D-mannose, sorbose, and the like;
  • disaccharides such as lactose, trehalose, and the like
  • cyclodextrins such as 2- hydroxypropyl-.beta.-cyclodextrin
  • polysaccharides such as raffinose, maltodextrins, dextrans, and the like
  • alditols such as mannitol, xylitol, and the like.
  • carbohydrates may includes lactose, threhalose, raffinose maltodextrins, and mannitol.
  • Suitable polypeptides include aspartame.
  • Amino acids include alanine and glycine, with glycine being used in some embodiments.
  • Additives which are minor components of the composition of this invention, may be included for conformational stability during spray drying and for improving dispersibility of the powder.
  • additives include hydrophobic amino acids such as tryptophan, tyrosine, leucine, phenylalanine, and the like.
  • Suitable pH adjusters or buffers include organic salts prepared from organic acids and bases, such as sodium citrate, sodium ascorbate, and the like; sodium citrate may be used in some embodiments.
  • Pulmonary administration of a micellar oligonucleotide conjugate formulation may be achieved through metered dose spray devices with propellants such as tetrafluoroethane, heptafluoroethane, dimethylfluoropropane, tetrafluoropropane, butane, isobutane, dimethyl ether and other non-CFC and CFC propellants.
  • propellants such as tetrafluoroethane, heptafluoroethane, dimethylfluoropropane, tetrafluoropropane, butane, isobutane, dimethyl ether and other non-CFC and CFC propellants.
  • compositions can be targeted to a surface of the oral cavity, e.g., to sublingual mucosa which includes the membrane of ventral surface of the tongue and the floor of the mouth or the buccal mucosa which constitutes the lining of the cheek.
  • the sublingual mucosa is relatively permeable thus giving rapid absorption and acceptable bioavailability of many drugs. Further, the sublingual mucosa is convenient, acceptable and easily accessible.
  • a pharmaceutical composition of oligonucleotide may also be administered to the buccal cavity of a human being by spraying into the cavity, without inhalation, from a metered dose spray dispenser, a mixed micellar pharmaceutical formulation as described above and a propellant.
  • the dispenser is first shaken prior to spraying the pharmaceutical formulation and propellant into the buccal cavity.
  • oligonucleotide agent can be disposed on or in a device, e.g., a device which implanted or otherwise placed in a subject.
  • Exemplary devices include devices which are introduced into the vasculature, e.g. , devices inserted into the lumen of a vascular tissue, or which devices themselves form a part of the vasculature, including stents, catheters, heart valves, and other vascular devices. These devices, e.g., catheters or stents, can be placed in the vasculature of the lung, heart, or leg.
  • Other devices include non-vascular devices, e.g. , devices implanted in the peritoneum, or in organ or glandular tissue, e.g. , artificial organs.
  • the device can release a therapeutic substance in addition to a oligonucleotide, e.g. , a device can release insulin.
  • Other devices include artificial joints, e.g. , hip joints, and other orthopedic implants.
  • unit doses or measured doses of a composition that includes oligonucleotide are dispensed by an implanted device.
  • the device can include a sensor that monitors a parameter within a subject.
  • the device can include pump, e.g., and, optionally, associated electronics.
  • Tissue e.g., cells or organs can be treated with An oligonucleotide agentex vivo and then administered or implanted in a subject.
  • the tissue can be autologous, allogeneic, or xenogeneic tissue.
  • tissue can be treated to reduce graft v. host disease.
  • the tissue is allogeneic and the tissue is treated to treat a disorder characterized by unwanted gene expression in that tissue.
  • tissue e.g., hematopoietic cells, e.g., bone marrow hematopoietic cells, can be treated to inhibit unwanted cell proliferation.
  • the oligonucleotide treated cells are insulated from other cells, e.g. , by a semi-permeable porous barrier that prevents the cells from leaving the implant, but enables molecules from the body to reach the cells and no molecules produced by the cells to enter the body.
  • the porous barrier is formed from alginate.
  • the invention features a method of administering an oligonucleotide agent.
  • the unit dose is less than 1 .4 mg per kg of bodyweight, or less than 10, 5, 2, 1 , 0.5, 0.1 , 0.05, 0.01 , 0.005, 0.001 , 0.0005, 0.0001 , 0.00005 or 0.00001 mg per kg of bodyweight, and less than 200 nmole of oligonucleotide agent (e.g., about 4.4. times.10.
  • sup.16 copies per kg of bodyweight, or less than 1500, 750, 300, 150, 75, 15, 7.5, 1 .5, 0.75, 0.15, 0.075, 0.015, 0.0075, 0.0015, 0.00075, 0.00015 nmole of RNA agent per kg of bodyweight.
  • the defined amount can be an amount effective to treat or prevent a disease or disorder, e.g., a disease or disorder associated with the target oligonucleotide.
  • the unit dose for example, can be administered by injection (e.g., intravenous or intramuscular), an inhaled dose, or a topical application. In some embodiments dosages may be less than 2, 1 , or 0.1 mg/kg of body weight.
  • the unit dose is administered less frequently than once a day, e.g., less than every 2, 4, 8 or 30 days.
  • the unit dose is not administered with a frequency (e.g., not a regular frequency).
  • the unit dose may be administered a single time.
  • the effective dose is administered with other traditional therapeutic modalities.
  • the subject has a viral infection and the modality is an antiviral agent other than an oligonucleotide agent.
  • the modality is an antiviral agent other than an oligonucleotide agent.
  • the patient can be monitored for changes in his condition and for alleviation of the symptoms of the disease state.
  • the dosage of the compound may either be increased in the event the patient does not respond significantly to current dosage levels, or the dose may be decreased if an alleviation of the symptoms of the disease state is observed, if the disease state has been ablated, or if undesired side-effects are observed.
  • intracapsular intracapsular
  • reservoir may be advisable.
  • oligonucleotide agent in conjunction with a known antiviral agent (e.g., a protease inhibitor or reverse transcriptase inhibitor).
  • a patient being treated for cancer may be administered an oligonucleotide agent in conjunction with a chemotherapy.
  • the concentration of the oligonucleotide agent composition is an amount sufficient to be effective in treating or preventing a disorder or to regulate a physiological condition in humans.
  • concentration or amount of oligonucleotide agent administered will depend on the parameters determined for the agent and the method of administration, e.g., nasal, buccal, pulmonary.
  • nasal formulations tend to require much lower concentrations of some ingredients in order to avoid irritation or burning of the nasal passages. It is sometimes desirable to dilute an oral formulation up to 10-100 times in order to provide a suitable nasal formulation.
  • Certain factors may influence the dosage required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the general health and/or age of the subject, and other diseases present.
  • treatment of a subject with a therapeutically effective amount of an oligonucleotide agent can include a single treatment or, for example, can include a series of treatments.
  • the effective dosage of a oligonucleotide agent such as a oligonucleotide agent used for treatment may increase or decrease over the course of a particular treatment. Changes in dosage may result and become apparent from the results of diagnostic assays as described herein.
  • the subject can be monitored after administering a oligonucleotide agent composition. Based on information from the monitoring, an additional amount of the oligonucleotide agent composition can be administered.
  • Dosing is dependent on severity and responsiveness of the disease condition to be treated, with the course of treatment lasting from several days to several months, or until a cure is effected or a diminution of disease state is achieved.
  • Optimal dosing schedules can be calculated from measurements of drug accumulation in the body of the patient. Persons of ordinary skill can easily determine optimum dosages, dosing methodologies and repetition rates. Optimum dosages may vary depending on the relative potency of individual compounds, and can generally be estimated based on EC50s found to be effective in in vitro and in vivo animal models.
  • oligonucleotide agents described herein can be administered to mammals, particularly large mammals such as nonhuman primates or humans in a number of ways.
  • the administration of the oligonucleotide agent intradermal, intraperitoneal, intramuscular, intrathecal, intraventricular, intracranial, subcutaneous, transmucosal, buccal, sublingual, endoscopic, rectal, oral, vaginal, topical, pulmonary, intranasal, urethral or ocular.
  • Administration can be provided by the subject or by another person, e.g., a health care provider.
  • the medication can be provided in measured doses or in a dispenser which delivers a metered dose. Selected modes of delivery are discussed in more detail below.
  • composition administered orally, e.g., in the form of tablets, capsules, gel capsules, lozenges, troches or liquid syrups. Further, the composition can be applied topically to a surface of the oral cavity.
  • oligonucleotide agents described herein can be any of the oligonucleotide agents described herein.
  • the medication can be sprayed into the buccal cavity or applied directly, e.g., in a liquid, solid, or gel form to a surface in the buccal cavity.
  • This administration is particularly desirable for the treatment of inflammations of the buccal cavity, e.g., the gums or tongue, e.g., in one embodiment, the buccal administration is by spraying into the cavity, e.g., without inhalation, from a dispenser, e.g., a metered dose spray dispenser that dispenses the pharmaceutical composition and a propellant.
  • the medications can be applied to the surface of the eye or nearby tissue, e.g., the inside of the eyelid. They can be applied topically, e.g., by spraying, in drops, as an eyewash, or an ointment. Administration can be provided by the subject or by another person, e.g., a health care provider.
  • the medication can be provided in measured doses or in a dispenser which delivers a metered dose.
  • the medication can also be administered to the interior of the eye, and can be introduced by a needle or other delivery device which can introduce it to a selected area or structure. Ocular treatment is particularly desirable for treating inflammation of the eye or nearby tissue.
  • the medication can be applied topically or delivered in a layer of the skin, e.g., by the use of a microneedle or a battery of
  • oligonucleotide agents described herein can be any of the oligonucleotide agents described herein.
  • Pulmonary administration can be achieved by inhalation or by the introduction of a delivery device into the pulmonary system, e.g., by introducing a delivery device which can dispense the medication.
  • Certain embodiments may use a method of pulmonary delivery by inhalation.
  • the medication can be provided in a dispenser which delivers the medication, e.g., wet or dry, in a form sufficiently small such that it can be inhaled.
  • the device can deliver a metered dose of medication.
  • the subject, or another person, can administer the medication.
  • Oligonucleotide agents can be formulated as a liquid or nonliquid, e.g., a powder, crystal, or aerosol for pulmonary delivery.
  • Nasal administration can be achieved by introduction of a delivery device into the nose, e.g., by introducing a delivery device which can dispense the medication.
  • Methods of nasal delivery include spray, aerosol, liquid, e.g., by drops, or by topical administration to a surface of the nasal cavity.
  • the medication can be provided in a dispenser with delivery of the medication, e.g., wet or dry, in a form sufficiently small such that it can be inhaled.
  • the device can deliver a metered dose of medication.
  • the subject, or another person, can administer the medication.
  • Nasal delivery is effective not only for disorders which directly affect nasal tissue, but also for disorders which affect other tissue
  • Oligonucleotide agents can be formulated as a liquid or nonliquid, e.g., a powder, crystal, or for nasal delivery.
  • An oligonucleotide agent can be packaged in a viral natural capsid or in a chemically or enzymatically produced artificial capsid or structure derived therefrom.
  • the dosage of a pharmaceutical composition including a
  • HBsAg has been shown to block many aspects of the immune response to HBV infection. Therefore, elimination of circulating HBsAg may be a critical factor in allowing the restoration of immunocompetence in patients with chronic hepatitis B infection.
  • An efficient method for eliminating HBsAg in the circulation is to prevent the formation and or release of subviral particles (SVPs) from infected cells (SVPs are the major carrier of HBsAg to the blood).
  • SVPs subviral particles
  • the morphogenesis and intracellular transit of SVPs can be modeled in vitro in BHK-21 cells by expressing the small form of the HBsAg protein
  • sHBsAg which is the form specifically enriched in SVPs.
  • This model system is considered to be a surrogate model for the morphogenesis and transit of HBV SVPs. Owing to the critical role of serum HBsAg in allowing chronicity of HBV infection, the efficacy of compounds in this model demonstrates their antiviral activity against HBV.
  • oligonucleotide conjugates can be introduced into BHK-21 cells at the same time as the template RNA for sHBsAg expression using electroporation. Activity in the BHK model system can be assessed by visualizing the location of HBsAg protein inside the BHK-21 cells by immunofluorescence microscopy. Formation of SVPs in the perinuclear space can be visualized by transmission electron microscopy. Oligonucleotide conjugates will be judged to be active if HBsAg is restricted to the perinuclear space and is prevented from transiting to the periphery of the cell (secretion) or if the formation of SVPs is prevented.
  • oligonucleotide conjugates Treatment of sHBsAg expressing BHK-21 cells with oligonucleotide conjugates as described herein is expected to demonstrate the ability of oligonucleotide conjugates to block formation of SVPs and the transit of sHBsAg in a sequence independent fashion.
  • the activity of oligonucleotide conjugates is expected to be dependent on the presence of phosphorothioation. Moreover, this ability is predicted to be retained in the presence of 2' ribose modification (such as for oligonucleotide conjugates that can be prepared with oligonucleotides comprising e.g. SEQ ID NOs 5, 6, 9 and/or 10) and base modification (5' methylcytosine, such as ONCs that can be prepared with oligonucleotides comprising e.g. SEQ ID nos. 3, 4, 9 and 10).
  • 2' ribose modification such as for oligonucleotide conjugates that can be
  • NAP conjugatess will be expected to be able to block the formation of and intracellular transit and secretion of SVPs from infected cells at oligonucleotide lengths from 20-120 nucleotides according to the sequence independent properties of NAPs as described in U.S. Pat. Nos. 8,008,269B2, 8,008,270B2 and 8,067,385B2.
  • Oligonucleotide-10-Ca is a calcium chelate complex of the an oligonucleotide coprising SEQ ID NO: 10 and can be prepared in normal saline using a ratio of 30 mg of CaCI 2 for every 100 mg of oligonucleotide present (see US20140065102 US20140065102).
  • the preparation of ON-1 as a calcium chelate complex can be used to improve the tolerability of ON administration (see International application publication no. WO2012/021985 and U.S. application publication no. 2012/0046348), and does not interfere with specific antiviral activity.
  • ON-10-Ca (which can be administered with weekly 500 mg doses) can clear HBsAg from the blood (and subsequently HBeAg) and HBV virions (HBV DNA) in HBV infected patients similarly to other ONs that do not comprise 2'-ribose modifications or modified bases (see Tables 3, 4 and 6 versus 7, 8 and 9 respectively of US20140065102).
  • the results disclosed in US20140065102 demonstrate that NAPs containing both 2' ribose modifications and modified bases (e.g. the ON referred to as REP 2139 of US20140065102) can act to reduce HBsAg in the blood and that ONs prepared as chelate complexes (e.g. the ON-chelate referred to as REP 2139-Ca in US20140065102) can be used to reduce or clear HBsAg from the blood.
  • ONs prepared as chelate complexes e.g. the ON-chelate referred to as REP 2139-Ca in US20140065
  • US20140065102 shows the effect of monotherapty with the ON- chelate referred to as REP 2139-Ca on serum HBsAg in patients with chronic HBV infection, (see Table 7 of US20140065102).
  • Interferon-based monotherapy typically results in only a moderate level ( ⁇ 50 mlU/ml) of anti-HBsAg antibody in the blood in a very small proportion of patients ( ⁇ 10%) after 48 weeks of therapy (Reijnders et al., 201 1 , J. Hepatol., 54: 449-454;
  • Harayiannis et al., 1990, J. Hepatol., 10: 350-352) and the antiviral effects of thymosin .alpha.1 are similarily limited (Yang et al., 2008, Antiviral Res. 77: 136-141 ).
  • Example III of US20140065102 also showed that the effects on the restoration of immune function occurred in all the patients tested, such that all who were tested achieved anti-HBsAg levels in the blood much more rapidly, and at much greater levels than observed with immunotherapy alone and in cases exceeding the levels of anti- HBsAg antibodies typically observed in healthy, non-infected HBsAg vaccinated individuals.
  • HBV transgenic mice (Guidotti LG, Matzke B, Schaller H, Chisari FV. High-level hepatitis B virus replication in transgenic mice. J Virol 1995 Oct 1 ; 69(10): 6158- 6169) were obtained from the Scripps Research Institute and housed at Utah State University where the treatment and sample collections were performed.
  • Blood was collected prior to dosing and on days 5, 9, and 14.
  • Serum samples were stored at -20°C and sent to GlaxoSmithKline, RTP for analysis. Serum HBsAg levels were determined using HBsAg One ELISA (International Immuno Diagnostics, Foster City, CA) according to the manufacturer's instructions.
  • HBsAg levels were normalized to the Day 1 (D1) value for each mouse and graphed as averages for the treatment group and for individual mice in each treatment group.
  • Figure 1 shows the average-fold change in Mean+SE levels in the HBV transgenic mice for the the saline vehicle along (black bars), the non-GalNAc -(AC)20 oligonucleotide (transparent bars) and the GalNac-conjugated -(AC)20 oligonucleotide (grey bars).
  • the average-fold change for the mean+SE level at D1 was the same for the saline vehicle, the non-GalNAc -(AC)20 oligonucleotide (SEQ ID NO: 10) and the GalNac -conjugated (AC)20 oligonucleotide (depicted in Scheme 1 ).
  • Figures 2A, 2B and 2C show the variability in fold change observed for individual HBV transgenic mice in the study for the saline vehicle alone (2A), the non-GalNac -(AC)20 oligonucleotide (2B) and the GalNac -cobjugated (AC)20 oligonucleotide (2C), respectively.
  • the HBsAg level changes for the vehicle group are typical for this transgenic mouse model.

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Abstract

L'invention concerne des conjugués oligonucléotidiques comprenant des conjugués oligonucléotidiques indépendants de séquences, comportant au moins une sous-unité de formule (I), ainsi que des chélates des conjugués oligonucléotidiques. L'invention concerne également des méthodes d'utilisation de ces conjugués oligonucléotidiques et chélates de conjugués oligonucléotidiques pour le traitement d'infections virales, notamment les infections par VHB ou les co-infections VHB/VHD, lesdites méthodes consistant à administrer au sujet un premier agent pharmaceutiquement acceptable qui est un conjugué oligonucléotidique ou son chélate, un éventuel second agent pharmaceutiquement acceptable qui stimule la fonction immunitaire et un éventuel troisième agent pharmaceutiquement acceptable comprenant un composé antiviral.
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CN111372593A (zh) * 2017-10-20 2020-07-03 戴瑟纳制药公司 治疗乙型肝炎感染的方法
US11166976B2 (en) 2018-11-08 2021-11-09 Aligos Therapeutics, Inc. S-antigen transport inhibiting oligonucleotide polymers and methods
US20220160748A1 (en) * 2020-11-20 2022-05-26 Aligos Therapeutics, Inc. Conjugates of s-antigen transport inhibiting oligonucleotide polymers having enhanced liver targeting
WO2024061157A1 (fr) * 2022-09-19 2024-03-28 Kylonova (Xiamen) Biopharma Co., Ltd. Conjugués glucide-oligonucléotide, compositions pharmaceutiques et applications thérapeutiques
EP4483906A1 (fr) * 2023-06-29 2025-01-01 SeeCure Taiwan Co., Ltd. Conjugué médicamenteux, composition pharmaceutique et procédé de traitement de l'hépatite
RU2853934C1 (ru) * 2023-06-29 2025-12-26 СиКьюэ Тайвань Ко., Лтд. Конъюгат лекарственного средства, фармацевтическая композиция и способ лечения гепатита

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US11166976B2 (en) 2018-11-08 2021-11-09 Aligos Therapeutics, Inc. S-antigen transport inhibiting oligonucleotide polymers and methods
US20220160748A1 (en) * 2020-11-20 2022-05-26 Aligos Therapeutics, Inc. Conjugates of s-antigen transport inhibiting oligonucleotide polymers having enhanced liver targeting
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