BR112013027046B1 - MODIFIED OLIGONUCLEOTIDE COMPOUNDS, THEIR USES IN MODULATING HEPATITIS B VIRUS (HBV) EXPRESSION AND PHARMACEUTICAL COMPOSITION THEREOF - Google Patents

Abstract

COMPOUND FOR MODULATION OF HEPATITIS B VIRUS (HBV) EXPRESSION, PHARMACEUTICAL COMPOSITION AND ITS USES IN THE TREATMENT OF INFECTION BY THE SAID VIRUS OR FOR REDUCING HBsAg LEVELS. The present invention relates to compounds comprising modified single-stranded oligonucleotide for modulating hepatitis B virus (HBV) expression. Furthermore, the present invention relates to a pharmaceutical composition comprising said compounds and the use of said compounds and said pharmaceutical composition in treating infection by said virus or for reducing HBsAg levels.

Description

Sequence Listing

[0001] This application is being deposited together with the Sequence Listing in electronic format. The sequence listing is provided as a file titled BIOL0175WOSEQ.txt created on April 18, 2012, which is approximately 256 KB in size. The information in electronic format of the Sequence Listing is incorporated herein by reference in its entirety.

Field

[0002] In certain embodiments, methods, compounds, and compositions are provided for inhibiting the expression of hepatitis B virus (HBV) mRNA and protein in an animal. Such methods, compounds and compositions are useful for treating, preventing or alleviating HBV related diseases and disorders.

Fundamentals

[0003] Hepatitis B is a viral disease transmitted parenterally through contaminated material, such as blood and blood products, contaminated needles, sexually and vertically from infected mothers or carriers to their offspring. It is estimated by the World Health Organization that more than 2 billion people are infected worldwide, with about 4 million acute cases per year, 1 million deaths per year, and 350-400 million chronic carriers (World Health Organization: Geographic Prevalence of Hepatitis B Prevalence, 2004. http://www.who.int/vaccines-surveillance/graphics/htmls/hepbprev.htm).

[0004] The virus, HBV is a double-stranded hepatotropic virus, which only infects humans and non-human primates. Viral replication occurs predominantly in the liver and, to a lesser extent, in the kidneys, pancreas, bone marrow and spleen ( Hepatitis B virus biology. Microbiol Mol Biol Rev. 64: 2000; 51-68 .). Viral and immune markers are detectable in the blood and characteristic antigen-antibody patterns evolve over time. The first detectable viral marker is HBsAg, followed by hepatitis B and antigen (HBeAg) and HBV DNA. Titers may be high during the incubation period, but HBV and HBeAg DNA levels begin to fall early in the illness and may be undetectable at the time of peak clinical illness (Hepatitis B Virus Infection—natural history and clinical consequences. N Engl J Med.. 350: 2004; 1118-1129). HBeAg is a viral marker detectable in blood and correlates with active viral replication and therefore high viral load and infectivity (Hepatitis B and antigen—the dangerous end game of hepatitis B. N Engl J Med. 347: 2002; 208-210). The presence of anti-HBsAb and anti-HBcAb (IgG) indicates recovery and immunity in a previously infected individual.

[0005] Currently, recommended therapies for chronic HBV infection from the American Association for the Study of Liver Diseases (AASLD) and the European Association for the Study of the Liver (EASL) include interferon alpha (INFα), pegylated interferonalpha-2a (Peg-IFN2a), entecavir, and tenofovir. Nucleoside and nucleotide therapies, entecavir and tenofovir, are successful in reducing viral load, but rates of HBeAg seroconversion and HBsAg loss are even lower than those obtained using IFNα therapy. Other similar therapies, including lamivudine (3TC), telbivudine (LdT), and adefovir are also used, but for nucleoside/nucleotide therapies in general, emergence of resistance limits therapeutic efficacy.

[0006] Thus, there is a need in the art to discover and develop new anti-viral therapies. Furthermore, there is a need for new anti-HBV therapies that can increase HBeAg and HBsAg seroconversion rates. Recent clinical research has found a correlation between seroconversion and reductions in HBeAg (Fried et al (2008) Hepatology 47:428) and reductions in HBsAg (Moucari et al (2009) Hepatology 49:1151). Reductions in antigen levels may have allowed immunological control of HBV infection, due to elevated levels of antigens believed to induce immune tolerance. Current nucleoside therapies for HBV are capable of dramatic reductions in serum HBV levels, but have little impact on HBsAg and HBeAg levels.

[0007] Antisense technology is emerging as an effective means to reduce the expression of specific gene products and therefore prove to be exceptionally useful in a number of therapeutic, diagnostic and research applications for modulating HBV expression (See US Patent Publication 2008/0039418 and 2007/0299027). Antisense therapy differs from nucleoside therapy in that it can directly target transcripts for HBV antigens and thereby reduce serum levels of HBeAg and HBsAg. Due to the multiple overlapping transcripts produced upon HBV infection, there is also an opportunity for a single antisense oligomer to reduce HBV DNA in addition to both HBeAg and HBsAg. Therefore, antisense technology is emerging as an effective means of reducing the expression of certain gene products and therefore may prove to be exceptionally useful in a number of therapeutic, diagnostic and research applications for HBV modulation.

summary

[0008] Provided herein are methods, compounds, and compositions for modulating HBV mRNA and protein expression. In certain embodiments, compounds useful for modulating HBV mRNA and protein expression are antisense compounds. In certain embodiments, the antisense compounds are antisense oligonucleotides.

[0009] In certain embodiments, the modulation may occur in a cell or tissue. In certain embodiments, the cell or tissue is from an animal. In certain embodiments, the animal is a human. In certain embodiments, HBV mRNA levels are reduced. In certain embodiments, levels of HBV DNA are reduced. In certain embodiments, HBV protein levels are reduced. In certain embodiments, HBV antigen levels are reduced. In certain embodiments, HBV antigen (HBsAg) levels are reduced. In certain embodiments, antigen and HBV (HBeAg) levels are reduced. This reduction can occur in a time-dependent manner, or in a dose-dependent manner.

[00010] Also provided are methods, compounds and compositions useful for the prevention, treatment and alleviation of diseases, disorders and conditions. In certain embodiments, these HBV-related disorders and conditions are liver diseases. In certain embodiments, these liver diseases, disorders, and conditions include jaundice, liver cancer, liver inflammation, liver fibrosis, inflammation, liver cirrhosis, liver failure, diffuse hepatocellular inflammatory disease, hemaphagocytic syndrome, serum hepatitis, HBV viremia, and transplant-related liver disease. In certain embodiments, these HBV-related diseases, disorders and conditions are hyperproliferative diseases, disorders and conditions. In certain embodiments such hyperproliferative diseases, disorders and conditions include cancer, as well as associated malignancies and metastases. In certain embodiments, such cancers include liver cancer and hepatocellular cancer (HCC).

[00011] Such diseases, disorders, and conditions may have one or more common risk factors, causes, or outcomes. Some risk factors and causes for developing liver disease or hyperproliferative disease include aging, tobacco use, exposure to sunlight and ionizing radiation; contact with certain chemicals; infection with certain viruses and bacteria; certain hormone therapies, family history of cancer; alcohol use, and certain lifestyle choices, including poor diet, lack of physical activity, and/or being overweight. Certain symptoms and outcomes associated with the development of liver disease or a hyperproliferative disease include, but are not limited to: flu-like illness, weakness, pain, headache, fever, loss of appetite, diarrhea, jaundice, nausea and vomiting, pain over the liver area of the body, clay-colored or gray stools, itching all over the body, and dark-colored urine.

[00012] In certain embodiments, the treatment methods include administering an HBV antisense compound to a subject in need thereof. In certain embodiments, methods of treatment include administering an HBV antisense oligonucleotide to a subject in need thereof.

Detailed Description

[00013] It should be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention as claimed. Here, the use of the singular includes the plural, unless otherwise noted. As used herein, the use of "or" means "and/or" unless otherwise noted. Furthermore, the use of the term "including" as well as other forms such as "include" and "includes" is not limiting. Furthermore, terms such as "element" or "component" encompass both elements and components comprising one unit and elements and components comprising more than one subunit, unless otherwise indicated.

[00014] Section titles used herein are for organizational purposes only and should not be construed as limiting the subject matter described. All documents, or portions of documents cited in this application, including, but not limited to, patents, patent applications, articles, books and treaties, are hereby expressly incorporated by reference to the portions of the document discussed herein, as well as in their entirety.

Definitions

[00015] Unless specific definitions are provided, the nomenclature used in connection with, and the procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well known and commonly used in the art. Standard techniques can be used for chemical synthesis and chemical analysis. Where permitted, all patents, applications, published applications and other publications, GENBANK Accession numbers and associated sequence information obtained through databases such as the National Center for Biotechnology Information (NCBI) and other data referred to throughout the disclosure herein are incorporated by reference to the portions of the document discussed herein, as well as in their entirety.

[00016] Unless otherwise indicated, the following terms have the following meanings:

[00017] "2'-O-methoxyethyl" (also 2'-MOE and 2'-O(CH2)2-OCH3) refers to an O-methoxy-ethyl modification at the 2' position of a furanose ring. A modified 2'-O-methoxyethyl sugar is a modified sugar.

[00018] The "2'-MOE nucleoside" (also 2'-O-methoxyethyl nucleoside) means a nucleoside comprising a modified 2'-MOE sugar moiety.

[00019] The "2'-substituted nucleoside" means a nucleoside comprising a substituent at the 2'-position of the furanosyl ring other than H or OH. In certain embodiments, 2'-substituted nucleosides include nucleosides with bicyclic sugar modifications.

[00020] The "3' target site" refers to the nucleotide of a target nucleic acid that is complementary to the 3'-maximal nucleotide of a particular antisense compound.

[00021] The "5' target site" refers to the nucleotide of a target nucleic acid that is complementary to the 5'-maximal nucleotide of a particular antisense compound.

[00022] "5-methylcytosine" means a cytosine modified with a methyl group attached to the 5 position. 5-Methylcytosine is a modified nucleobase.

[00023] "About" means within ±7% of a value. For example, if it is stated, "compounds effected at least about 70% inhibition of HBV", it is implied that HBV levels are inhibited within a range of 63% and 77%.

[00024] "Acceptable safety profile" means a pattern of side effects that is within clinically acceptable limits.

[00025] "Active pharmaceutical agent" means the substance or substances in a pharmaceutical composition that provide a therapeutic benefit when administered to an individual. For example, in certain embodiments a target antisense oligonucleotide for HBV is an active pharmaceutical agent.

[00026] "Active target region" means a target region to which one or more active antisense compounds is targeted. "Active antisense compounds" means antisense compounds that reduce target nucleic acid levels or protein levels.

[00027] "Acute hepatitis B infection" results when a person exposed to the hepatitis B virus begins to develop the signs and symptoms of viral hepatitis. This period of time, called the incubation period, averages 90 days, but can be as short as 45 days or as long as 6 months. For most people this infection will cause mild to moderate discomfort, but it will go away on its own as the body's immune response is able to fight off the virus. However, some people, particularly those with compromised immune systems such as people suffering from AIDS, undergoing chemotherapy, taking immunosuppressant drugs, or taking steroids, have many serious problems as a result of acute HBV infection and progress to more severe conditions such as fulminant liver failure.

[00028] "Concurrently administered" refers to the co-administration of two agents in any form in which the pharmacological effects of both are manifested in the patient at the same time. Co-administration does not require that both agents be administered in a single pharmaceutical composition, in the same dosage form, or by the same route of administration. The effects of both agents need not manifest at the same time. Effects only need to overlap for a period of time and need not be coextensive.

[00029] "Administer" means providing a pharmaceutical agent to an individual, and includes, but is not limited to, administration by a medical professional and self-administration.

[00030] "Agent" means an active substance that can provide a therapeutic benefit when administered to an animal. "First agent" means a therapeutic compound described herein. For example, a first agent can be an antisense oligonucleotide targeting HBV. "Second agent" means a second therapeutic compound described herein (e.g., a second HBV-targeted antisense oligonucleotide) and/or a non-HBV therapeutic compound.

[00031] "Improvement" refers to a decrease in at least one indicator of the severity of a condition or disease. The severity of the indicators can be determined by subjective or objective measures, which are known to those skilled in the art.

[00032] "Animal" refers to a human or non-human animal, including, but not limited to, mice, rats, rabbits, dogs, cats, pigs, and non-human primates, including, but not limited to, monkeys and chimpanzees.

[00033] "Antibody" refers to a molecule characterized by reacting specifically with an antigen in some way, where antibody and antigen are each defined in terms of the other. Antibody can refer to a complete antibody molecule or any fragment or region thereof, such as the heavy chain, light chain, Fab region, and Fc region.

[00034] "Antisense activity" means any detectable or measurable activity attributable to the hybridization of an antisense compound to its target nucleic acid. In certain embodiments, antisense activity is a decrease in the amount or expression of a target nucleic acid or protein encoded by this target nucleic acid.

[00035] "Antisense compound" means an oligomeric compound that is capable of undergoing hybridization with a target nucleic acid through hydrogen bonding. Examples of antisense compounds include single-stranded and double-stranded compounds such as antisense oligonucleotides, siRNAs, shRNAs, snoRNAs, miRNAs, and satellite repeats.

[00036] "Antisense inhibition" means the reduction of target nucleic acid levels in the presence of an antisense compound complementary to a target nucleic acid compared to target nucleic acid levels in the absence of the antisense compound.

[00037] "Antisense mechanisms" are all those mechanisms involving the hybridization of a compound to the target nucleic acid, where the result or effect of the hybridization is either target degradation or target occupation concomitant with the jamming of cellular machinery involving, for example, transcription or splicing.

[00038] "Antisense oligonucleotide" means a single-stranded oligonucleotide having a nucleobase sequence that allows hybridization to a corresponding region or segment of a target nucleic acid.

[00039] "Base complementarity" refers to the ability of precise base pairing of nucleobases of an antisense oligonucleotide with corresponding nucleobases in a target nucleic acid (i.e., hybridization), and is mediated by Watson-Crick, Hoogsteen, or inverted Hoogsteen hydrogen bonding between corresponding nucleobases.

[00040] "Bicyclic sugar" means a furanose ring modified by the bridge of two non-geminal carbon atoms. A bicyclic sugar is a modified sugar.

[00041] "Body weight" refers to the entire body weight of an animal, including all tissues, including adipose tissue.

[00042] "Caping structure" or "terminal capping portion" means chemical modifications, which have been incorporated into any terminal of an antisense compound.

[00043] "cEt" or "restricted ethyl" means a bicyclic sugar molecule comprising a bridge connecting the 4'-carbon and the 2'-carbon, wherein the bridge has the formula: 4'-CH(CH3)-O-2'.

[00044] "Restricted ethyl nucleoside" (also cEt nucleoside) means a nucleoside comprising a bicyclic sugar moiety comprising a 4'-CH(CH3)-O-2' bridge.

[00045] "Chemically distinct region" refers to a region of an antisense compound that is in some way chemically different from another region of the same antisense compound. For example, a region having 2'-O-methoxyethyl nucleotides is chemically distinct from a region having unmodified 2'-O-methoxyethyl nucleotides.

[00046] "Chimeric antisense compounds" means antisense compounds that have at least two chemically distinct regions, each position having a plurality of subunits.

[00047] "Chronic hepatitis B infection" occurs when a person initially suffers from an acute infection but is unable to fight off the infection. Whether the disease becomes chronic or resolves completely depends mainly on the age of the infected individual. About 90% of babies infected at birth will progress to chronic disease. However, as a person ages, the risk of chronic infection decreases such that between 20% to 50% of children and less than 10% of older children or adults will progress from acute to chronic infection. Chronic HBV infections are the primary treatment goal of the embodiments of the present invention, although the ASO compositions of the present invention are also capable of treating HBV-related conditions, such as inflammation, fibrosis, cirrhosis, liver cancer, serum hepatitis, and more.

[00048] "Coadministration" means the administration of two or more pharmaceutical agents to an individual. The two or more pharmaceutical agents can be in a single pharmaceutical composition, or they can be in separate pharmaceutical compositions. Each of the two or more pharmaceutical agents can be administered by the same or different routes of administration. Co-administration encompasses administration in parallel or sequentially.

[00049] "Complementarity" means the ability to pair between the nucleobases of a first nucleic acid and a second nucleic acid.

[00050] "Satisfy" means adherence to a therapy recommended by an individual.

[00051] "Comprises", "comprises" and "comprising" shall be understood to imply the inclusion of an indicated element or step, or group of steps or elements, but not the exclusion of any other element or step or group of steps or elements.

[00052] "Contiguous nucleobases" means nucleobases immediately adjacent to each other.

[00053] "Cure" means a method or course that restores health or a prescribed treatment for an illness.

[00054] "Deoxyribonucleotide" means a nucleotide having a hydrogen at the 2' position of the sugar portion of the nucleotide. Deoxyribonucleotides can be modified with any of a variety of substituents.

[00055] "Designing" or "Designing" refers to the process of designing an oligomeric compound that specifically hybridizes with a selected nucleic acid molecule.

[00056] "Diluent" means an ingredient in a composition that has no pharmacological activity, but is pharmaceutically necessary or desirable. For example, in drugs that are injected, the diluent can be a liquid, for example saline.

[00057] "Dosage unit" means a form in which a pharmaceutical agent is provided, for example, pill, tablet or other dosage unit known in the art.

[00058] "Dose" means a specified amount of a pharmaceutical agent delivered in a single administration, or over a specified period of time. In certain embodiments, a dose can be administered as two or more boluses, pills, or injections. For example, in certain embodiments, when subcutaneous administration is desired, the desired dose requires a volume that is not easily accommodated by a single injection. In such embodiments, two or more injections can be used to achieve the desired dose. In certain embodiments, a dose can be administered in two or more injections to minimize injection site reaction in an individual. In other embodiments, the pharmaceutical agent is administered by infusion over an extended period of time or continuously. Doses can be indicated as the amount of pharmaceutical agent per hour, day, week or month.

[00059] "Dosage regimen" is a combination of doses designed to achieve one or more desired effects.

[00060] "Duration" means the period of time during which an activity or event continues. In certain embodiments, the duration of treatment is the period of time over which doses of a pharmaceutical agent are administered.

[00061] "Effective amount", in the context of modulating an activity or for the treatment or prevention of a condition means the administration of that amount of active ingredient to a subject in need of such modulation, treatment or prophylaxis, either in a single dose or as part of a series, which is effective for modulating the effect, or for the treatment or prophylaxis or amelioration of such condition. The effective amount will vary depending on the physical condition and health of the subject being treated, the taxonomic group of subjects being treated, formulation of the composition, assessment of the medical situation, and other relevant factors.

[00062] "Efficacy" means the ability to produce the desired effect.

[00063] "Expression" includes all functions by which the encoded information of a gene is converted into structures present and operating in a cell. These structures include, but are not limited to, transcription and translation products.

[00064] "Fully complementary" or "100% complementary" means that each nucleobase of a first nucleic acid has a complementary nucleobase in a second nucleic acid. In certain embodiments, a first nucleic acid is an antisense compound and a target nucleic acid is a second nucleic acid.

[00065] "Fully modified motif" refers to an antisense compound comprising a contiguous sequence of nucleotides, wherein essentially each nucleoside is a sugar-modified nucleoside having uniform modification.

[00066] "Gapmer" means a chimeric antisense compound in which an inner region having a plurality of nucleosides that support RNase H cleavage is positioned between outer regions having one or more nucleosides wherein the nucleosides comprising the inner region are chemically distinct from the nucleoside or nucleosides comprising the outer regions. The inner region can be referred to as the "gap" and the outer regions can be referred to as the "wings".

[00067] "Amplified gap" means an antisense compound having a gap segment of 12 or more contiguous 2' deoxyribonucleotides positioned between the 5' and 3' wing segments having from one to six nucleotides having modified sugar moieties.

[00068] "HBV" means mammalian hepatitis B virus including human hepatitis B virus. The term encompasses geographic genotypes of hepatitis B virus, particularly human hepatitis B virus, as well as variant strains of geographic genotypes of hepatitis B virus.

[00069] "HBV antigen" means any hepatitis B virus antigen or protein, including core proteins such as "hepatitis B core antigen" or "HBcAg" and "hepatitis B E antigen" or "HBeAG" and envelope proteins such as "HBV surface antigen" or "HBsAg" or "HBsAG".

[00070] "Hepatitis B E antigen" or "HBeAg" or "HBeAG" is a non-particulate, secreted form of HBV core protein. HBV HBeAg and HBcAg antigens share primary amino acid sequences, so they show cross-reactivity at the T-cell level. HBeAg is not required for viral assembly or replication, although studies suggest that the establishment of chronic infection may be necessary. Neonatal infection with HBeAg-negative mutants often results in acute fulminant HBV infection rather than chronic HBV infection (Terezawa et al (1991) Pediatr. Res. 29:5), whereas infection of young woodchucks with WHeAg-negative mutants results in a much lower rate of chronic WHV infection (Cote et al (2000) Hepatology 31:190). HBeAg may eventually function as a tolerogen by inactivating specific T cells through nuclear deletion or clonal anergy ( Milich et al (1998) J. Immunol. 160:8102 ). There is a positive correlation between a reduction in HBV viral load and antigens and a reduction in T cell expression of inhibitory receptor programmed death-1 (PD-1, also known as PDCD1), a negative regulator of activated T cells, upon antiviral therapy and HBeAg seroconversion (Evans et al (2008) Hepatology 48:759).

[00071] "HBV mRNA" means any messenger RNA expressed by the hepatitis B virus.

[00072] "HBV nucleic acid" or "HBV DNA" means any nucleic acid encoding HBV. For example, in certain embodiments, an HBV nucleic acid includes, without limitation, any viral DNA sequence encoding an HBV genome or a portion thereof, any RNA sequence transcribed from a viral DNA including any mRNA sequence encoding an HBV protein.

[00073] "HBV protein" means any protein secreted by the hepatitis B virus. The term encompasses various HBV antigens, including core proteins such as "hepatitis E antigen, "HBeAg" or "HBeAG", and envelope proteins such as "HBV surface antigen" or "HBsAg".

[00074] "HBV surface antigen", or "HBsAg", or "HBsAG" is the envelope protein of infectious HBV viral particles, but it is also secreted as a non-infectious particle, with serum levels 1000 times higher than HBV viral particles. Serum levels of HBsAg in an infected person or animal can be as high as 1000 µg/mL mL (Kann and Gehrlich (1998) Topley & Wilson's Microbiology and Microbial Infections, 9th ed. 745). In acute HBV infections, the half-life of serum HBsAg, or serum t^, is 8.3 days ( Chulanov et al (2003) J. Med. Virol. 69: 313 ). The uptake of HBsAg by myeloid dendritic cells inhibits the upregulation of costimulatory molecules (i.e., B7) and inhibits the stimulatory capacity of T cells (den Brouw et al (2008) Immunology 126:280), and dendritic cells from chronically infected patients also show deficits in the expression of costimulatory molecules, IL-12 secretion, and T cell stimulation in the presence of HB sAg ( Zheng et al (2004) J. Viral Hepatitis 11:217 ). HBsAg-specific CD8 cells from CHB patients show altered tetramer binding. These CD8 cells are not anergic, but may have TCR topology that confers tolerance or partial ignorance (Reignat et al (2002) J. Exp. Med. 195:1089). In addition, reduction in serum HBsAg > log 1 at week 24 has a high predictive value (92%) for sustained virological response (SVR - defined as HBV DNA undetectable by PCR at 1 year after treatment) during Peg therapy -IFNα2a ( Moucari et al (2009) Hepatology 49:1151 ).

[00075] "Hepatitis B-related condition" or "HBV-related condition" means any disease, biological condition, medical condition, or event that is exacerbated, caused by, related to, associated with, or traceable to, a hepatitis B infection, exposure, or illness. Diffuse hepatocellular inflammatory disease, hemophagocytic syndrome, serum hepatitis, HBV viremia, transplantation-related liver disease, and conditions that have symptoms that may include one or all of the following: flu-like illness, weakness, aches, headaches, fever, loss of appetite, diarrhea, nausea and vomiting, pain along the liver area of the body, clay-colored or gray stools, itching all over the body, and dark-colored urine, when coupled with a positive test for the presence of a hepatitis B virus, an antigen hepatitis B viral infection, or a positive test for the presence of an antibody specific to a viral hepatitis B antigen.

[00076] "Hybridization" means the annealing of complementary nucleic acid molecules. In certain embodiments, complementary nucleic acid molecules include, but are not limited to, an antisense compound and a target nucleic acid. In certain embodiments, the complementary nucleic acid molecules include, but are not limited to, an antisense oligonucleotide and a target nucleic acid.

[00077] "Identifying an animal having an HBV infection" means identifying an animal having been diagnosed with HBV; or, identification of an animal having any symptom of an HBV infection, including, but not limited to, chronic HBV infection, inflammation, fibrosis, cirrhosis, liver cancer, serum hepatitis, jaundice, liver cancer, liver inflammation, liver fibrosis, liver cirrhosis, liver failure, diffuse hepatocellular inflammatory disease, hematophagocytic syndrome, serum hepatitis, HBV viremia, transplantation-related liver disease, and conditions that have symptoms that may include any or all of the following: disease flu-like weakness, aches, headaches, fever, loss of appetite, diarrhea, nausea and vomiting, pain over the liver area of the body, clay-colored or gray stools, itching all over the body, and dark-colored urine, when coupled with a positive test for the presence of a hepatitis B virus, a hepatitis B viral antigen, or a positive test for the presence of an antibody specific to a hepatitis B viral antigen, when coupled with a positive test for the presence of a hepatitis B virus, a hepatitis B viral antigen, hepatitis B, or a positive test for the presence of an antibody specific for a hepatitis B viral antigen.

[00078] "Immediately adjacent" means that there are no intervening elements between immediately adjacent elements.

[00079] "Subject" means a human or non-human animal selected for treatment or therapy.

[00080] "Individual adherence" means adherence by an individual to a recommended or prescribed treatment.

[00081] "Induce", "inhibit", "potentiate", "elevate", "increase", "decrease" or similar, generally denote quantitative differences between the two states. These terms can refer to a statistically significant difference between the two states. For example, "an amount effective to inhibit HBV activity or expression" means that the level of HBV activity or expression in a treated sample will differ quantitatively, and may be statistically significant, from the level of HBV activity or expression in untreated cells. Such terms are applied to, for example, levels of expression and levels of activity.

[00082] "HBV inhibition" means reducing the level or expression of an HBV mRNA, DNA and/or protein. In certain embodiments, HBV is inhibited in the presence of an HBV target antisense compound, including an HBV target antisense oligonucleotide, as compared to expression levels of HBV mRNA, DNA, and/or proteins in the absence of an HBV antisense compound, such as an antisense oligonucleotide.

[00083] "Inhibition of expression or activity" refers to a reduction, blockage of expression or activity and does not necessarily indicate a complete elimination of expression or activity.

[00084] "Injection site reaction" means abnormal inflammation or redness of the skin at the site of an injection in an individual.

[00085] "Internucleoside linkage" refers to the chemical linkage between nucleosides.

[00086] "Intraperitoneal administration" means administration by infusion or injection into the peritoneum.

[00087] "Intravenous administration" means administration into a vein.

[00088] "Extended" antisense oligonucleotides are those that have one or more additional nucleosides relative to an antisense oligonucleotide described herein.

[00089] "Linked deoxynucleoside" means a nucleic acid base (A, G, C, T, U) substituted for deoxyribose linked with a phosphate ester to form a nucleotide.

[00090] "Linked nucleosides" means adjacent nucleosides linked together by an internucleoside bond.

[00091] "Locked nucleic acid" or "LNA" or "LNA nucleosides" means nucleic acid monomers with a connecting bridge between two carbon atoms between the 4' and 2' position of the nucleoside sugar unit, thus forming a bicyclic sugar. Examples of this type of bicyclic sugar include, but are not limited to: A) α-L-Methyleneoxy(4'-CH2-O-2') LNA, (B) β-D-Methyleneoxy(4'-CH2-O-2') LNA, (C) Ethyleneoxy(4'-(CH2)2-O-2') LNA, (D) Aminooxy(4'-CH2-ON(R)-2') LNA, and (E) Oxyamino (4'-CH2-N(R)-O-2') LNA, as depicted below.

[00092] As used herein, LNA compounds include, but are not limited to, compounds having at least one bridge between the 4' and 2' position of the sugar wherein each bridge independently comprises 1 or 2 to 4 independently linked groups selected from -[C(R1)(R2)]n-, -C(R1)=C(R2)-, -C(R1)=N-, -C(=NR1)-, -C(=O)- , -C(=S)-, -O-, -Si(R1)2-, -S(=O)x- and -N(R1)-; where: x is 0, 1, or 2; n is 1, 2, 3, or 4; each of R1 and R2 is, independently, H, a protecting group, hydroxyl, C1-C12 alkyl, substituted C1-C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkylkynyl, C5-C20 aryl, C5-C20 substituted aryl, a heterocyclic radical, a substituted heterocyclic radical, hetero aryl, substituted heteroaryl, C5-C7 acyclic radical, C5-C7 substituted acyclic radical, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)2-J1), or sulfoxyl (S(=O)-J1); and each of J1 and J2 is independently H, C1-C12 alkyl, substituted C1-C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkyl, C5-C20 aryl, C5-C20 substituted aryl, acyl (C(=O)-H), substituted acyl, a heterocyclic radical, a substituted heterocyclic, C1-C12 aminoalkyl, substituted C1-C12 aminoalkyl radical or a protecting group.

[00093] Examples of 4'-2' bridging groups encompassed within the definition of LNA include, but are not limited to, one of the formulas: -[C(Ri)(R2)]n-, -[C(Ri)(R2)]n-O-, -C(RIR2)-N(RI)-0- or —C(R1R2)-O-N(R1)-. In addition, other bridge groups encompassed by the LNA definition are 4'-CH2-2', 4'-(CH2)2-2', 4'-(CH2)3-2', 4'-CH2-O-2', 4'-(CH2)2-O-2', 4'-CH2-O-N(R1)-2', and 4'-CH2-N(R1)-O-2'- bridges, where each R 1 and R2 is independently H, a protecting group or C1-C12 alkyl.

[00094] Also included within the definition of LNA according to the invention are LNAs in which the 2'-hydroxyl group of the ribosyl sugar ring is connected to the 4' carbon atom of the sugar ring, thereby forming a methylene-oxy bridge (4'-CH2-O-2') to form the bicyclic sugar moiety. The bridge can also be a methylene (-CH2-) group connecting the 2' oxygen atom and the 4' carbon atom, for which the term methylene-oxy (4'-CH2-O-2') LNA is used. Furthermore; in the case of the bicyclic sugar moiety having an ethylene bridging group at this position, the term ethyleneoxy(4'-CH2CH2-O-2') LNA is used. α -L-methylene-oxy (4'-CH2-O-2'), an isomer of methylene-oxy (4'-CH2-O-2') LNA is also encompassed within the definition of LNA, as used herein.

[00095] "Mismatch" or "non-complementary nucleobase" refers to the case where a nucleobase of a first nucleic acid is not capable of pairing with the corresponding nucleobase of a second or target nucleic acid.

[00096] "Modified internucleoside linkage" refers to a replacement or any alteration of a naturally occurring internucleoside linkage (i.e., a phosphodiester internucleoside linkage).

[00097] "Modified nucleobase" means any nucleobase except adenine, cytosine, guanine, thymidine or uracil. An "unmodified nucleobase" means the purine bases adenine (A) and guanine (G) and pyrimidine bases thymine (T), cytosine (C) and uracil (U).

[00098] "Modified nucleoside" means a nucleoside independently having a modified sugar and/or modified nucleobase moiety.

[00099] "Modified nucleotide" means a nucleotide independently having a modified sugar moiety, modified internucleoside linkage, or modified nucleobase.

[000100] "Modified oligonucleotide" means an oligonucleotide comprising at least one modified internucleoside linkage, a modified sugar, and/or a modified nucleobase.

[000101] "Modified sugar" means the replacement and/or any alteration of a portion of natural sugar.

[000102] "Monomer" refers to a single unit of an oligomer. Monomers include, but are not limited to, nucleosides and nucleotides, whether naturally occurring or modified.

[000103] "Motif" means the pattern of unmodified and modified nucleosides in an antisense compound.

[000104] "Natural sugar moiety" means a sugar molecule found in DNA (2'-H) or RNA (2'-OH).

[000105] "Natural-occurring internucleoside linkage" means a 3' to 5' phosphodiester linkage.

[000106] "Non-complementary nucleobase" refers to a pair of nucleobases that do not form hydrogen bonds with each other or otherwise support hybridization.

[000107] "Nucleic acid" refers to molecules composed of monomeric nucleotides. A nucleic acid includes, but is not limited to, ribonucleic acids (RNA), deoxyribonucleic acids (DNA), single-stranded nucleic acids, double-stranded nucleic acids, small interfering ribonucleic acids (siRNA), and microRNAs (miRNA).

[000108] "Nucleobase" means a heterocyclic radical capable of pairing with a base another nucleic acid.

[000109] "Nucleobase complementarity" refers to a nucleobase that is capable of base pairing with another nucleobase. For example, in DNA, adenine (A) is complementary to thymine (T). For example, in RNA, adenine (A) is complementary to uracil (U). In certain embodiments, complementary nucleobases refer to a nucleobase of an antisense compound that is capable of base pairing with a nucleobase of its target nucleic acid. For example, if a nucleobase at a particular position of an antisense compound is capable of hydrogen bonding with a nitrogenous base at a particular position of a target nucleic acid, then the position of hydrogen bonding between the oligonucleotide and the target nucleic acid is considered complementary at that nucleobase pair.

[000110] "Nucleobase sequence" means the end of contiguous nucleobases independent of any sugar, linkage, and/or nucleobase modification.

[000111] "Nucleoside" means a nucleobase attached to a sugar.

[000112] "Nucleoside mimetic" includes those structures used to replace the sugar or the sugar and the base, and not necessarily binding to one or more positions of an oligomeric compound, such as, for example, nucleoside mimetics with morpholino, cyclohexenyl, cyclohexyl, tetrahydropyranyl, bicycle or tricycle sugar mimetics, for example, non-furanose sugar units. Mimetic nucleotides include those structures used to replace the nucleoside and linkage at one or more positions of an oligomeric compound, for example, peptide nucleic acids or Morpholinos (morpholinos linked by -N(H)-C(=O)-O- or other non-phosphodiester linkages). Sugar substitute overlaps with nucleoside mimetic in the somewhat broader term, but is intended to indicate replacement of the sugar unit (furanose ring) only. The tetrahydropyranyl rings provided herein are illustrative of an example of a sugar substitute where the furanose sugar group has been replaced with a tetrahydropyranyl ring system. "Mimetic" refers to groups that are substituted for a sugar, a nucleobase, and/or internucleoside linkage. Generally, a mimetic is used in place of the sugar or internucleoside-sugar linkage combination, and the nucleobase is retained for hybridization with a selected target.

[000113] "Nucleotide" means a nucleoside having a phosphate group covalently attached to the sugar moiety of the nucleoside.

[000114] "Off-target effect" refers to an undesirable or harmful biological effect related to the modulation of RNA or protein expression of a gene other than the intended target nucleic acid.

[000115] "Oligomeric compound" means a polymer of linked monomeric subunits that is capable of hybridizing to at least one region of a nucleic acid molecule.

[000116] "Oligonucleoside" means an oligonucleotide in which the internucleoside linkages do not contain a phosphorus atom.

[000117] "Oligonucleotide" means a polymer of linked nucleotides, each of which may or may not be modified, independently of each other.

[000118] "Parenteral administration" means administration by injection (eg, bolus injection) or infusion. Parenteral administration includes subcutaneous administration, intravenous administration, intramuscular administration, intraarterial administration, intraperitoneal administration, or intracranial administration, for example, intracerebroventricular or intrathecal administration.

[000119] "Peptide" means a molecule formed by linking at least two amino acids by amide bonds. Without limitation, as used herein, "peptide" refers to both polypeptides and proteins.

[000120] "Pharmaceutically acceptable carrier" means a medium or diluent that does not interfere with the structure of the oligonucleotide. Some of these vehicles allow pharmaceutical compositions to be formulated as, for example, tablets, pills, dragees, capsules, liquids, gels, syrups, pastes, suspensions and lozenges for oral ingestion by a subject.

[000121] "Pharmaceutically acceptable derivative" includes pharmaceutically acceptable salts, prodrugs, and conjugates or isomers of the compounds described herein.

[000122] "Pharmaceutically acceptable salts" means physiologically and pharmaceutically acceptable salts of antisense compounds, that is, salts that retain the desired biological activity of the source oligonucleotide and do not confer undesirable toxicological effects on it.

[000123] "Pharmaceutical agent" means a substance that provides a therapeutic benefit when administered to an individual. For example, in certain embodiments, an HBV-targeted antisense oligonucleotide is a pharmaceutical agent.

[000124] "Pharmaceutical composition" means a mixture of substances suitable for administration to a subject. For example, a pharmaceutical composition can comprise an antisense oligonucleotide and a sterile aqueous solution. In certain embodiments, a pharmaceutical composition exhibits activity in the free uptake assay in certain cell lines.

[000125] "Phosphorothioate linkage" means a linkage between nucleosides in which the phosphodiester linkage is modified by replacing one of the non-bridge oxygen atoms with a sulfur atom. A phosphorothioate linkage is a modified internucleoside linkage.

[000126] "Portion" means a defined number of contiguous (i.e. linked) nucleobases of a nucleic acid. In certain embodiments, a moiety is a defined number of contiguous nucleobases of a target nucleic acid. In certain embodiments, a moiety is a defined number of contiguous nucleobases of an antisense compound.

[000127] "Prevention" or "preventing" refers to preventing or delaying the onset or development of a condition or disease over a period of time from hours to days, preferably weeks to months.

[000128] "Prodrug" means a therapeutic agent that is prepared in an inactive form that is converted into an active form (eg drug) within the body or cells thereof by the action of endogenous enzymes or other chemicals and/or conditions.

[000129] "Prophylactically effective amount" refers to an amount of a pharmaceutical agent, which provides a prophylactic or preventive benefit to an animal.

[000130] "Recommended therapy" means a therapeutic regimen recommended by a physician for the treatment, alleviation, or prevention of a disease.

[000131] "Region" is defined as a portion of the target nucleic acid with at least one identifiable structure, function or feature.

[000132] "Ribonucleotide" means a nucleotide having a hydroxy group at the 2' position of the sugar portion of the nucleotide. Ribonucleotides can be modified with any of a variety of substituents.

[000133] "Salts" means physiologically and pharmaceutically acceptable salts of the antisense compounds, that is, salts that retain the desired biological activity of the original oligonucleotide and do not confer undesirable toxicological effects on it.

[000134] "Segments" are defined as smaller portions or subportions of regions within a target nucleic acid.

[000135] "Seroconversion" is defined as the absence of serum HbeAg plus the presence of serum HBeAb, if it controls HBeAg as a determinant for seroconversion, or defined as the absence of serum HBsAg, if it controls HBsAg as a determinant for seroconversion, as determined by limits of currently available commercial ELISA detection systems.

[000136] "Shortened" or "truncated" versions of antisense oligonucleotides taught here have one, two, or more nucleosides deleted.

[000137] "Side Effects" means physiological responses attributable to a treatment other than the desired effects. In certain embodiments, side effects include, without limitation, injection site reactions, liver function test abnormalities, kidney function abnormalities, liver toxicity, renal toxicity, central nervous system abnormalities, and myopathies. For example, increased serum aminotransferase levels may indicate liver toxicity or abnormal liver function. For example, increased bilirubin may indicate liver toxicity or abnormal liver function.

[000138] "Significant", as used herein, means measurable or observable, for example, a significant result, such as a significant improvement or significant reduction, generally refers to a measurable or observable result, such as a measurable or observable improvement or reduction.

[000139] "Sites", as used herein, are defined as unique nucleobase positions within a target nucleic acid.

[000140] "Delays progression" means a decrease in the development of said disease.

[000141] "Specifically hybridizable" refers to an antisense compound having a sufficient degree of complementarity between an antisense oligonucleotide and a target nucleic acid to induce a desired effect, while exhibiting minimal or no effect on non-target nucleic acids, under conditions where specific binding is desired, i.e., under physiological conditions in the case of in vivo assays and therapeutic treatments. "Stringent hybridization conditions" or "stringent conditions" refer to conditions under which an oligomeric compound will hybridize to its target sequence, but to a minimal number of other sequences.

[000142] "Statistically significant" as used herein means a measurable or observable parameter that is unlikely to occur by chance.

[000143] "Subcutaneous administration" means administration just under the skin.

[000144] "Subject" means a human or non-human animal selected for treatment or therapy.

[000145] "Target" refers to a protein, the modulation of which is desired.

[000146] "Target gene" refers to a gene that encodes a target.

[000147] "Targeting" means the process of designing and selecting an antisense compound that will specifically hybridize with a target nucleic acid and induce a desired effect.

[000148] "Target nucleic acid", "target RNA", "target RNA transcript" and "target nucleic acid" all mean a nucleic acid capable of being targeted by antisense compounds.

[000149] "Target region" means a portion of a target nucleic acid where one or more antisense compounds are targeted.

[000150] "Target segment" means the nucleotide sequence of a target nucleic acid in which an antisense compound is targeted. "5' target site" refers to the 5'-maximum nucleotide of a target segment. "3' target site" refers to the 3'-maximum nucleotide of a target segment.

[000151] "Therapeutically effective amount" means an amount of a pharmaceutical agent that provides a therapeutic benefit to an individual.

[000152] "Treatment" refers to the administration of a composition to effect a change or amelioration of the disease or condition.

[000153] "Unmodified" nucleobases means the purine bases adenine (A) and guanine (G) and pyrimidine bases thymine (T), cytosine (C) and uracil (U).

[000154] "Unmodified nucleotide" means a nucleotide composed of naturally occurring nucleobases, sugar moieties and internucleoside linkages. In certain embodiments, an unmodified nucleotide is an RNA nucleotide (i.e., β-D-ribonucleosides) or a DNA nucleotide (i.e., D-β-deoxyribonucleosides).

[000155] "Validated target segment" is defined as at least a portion of 8 nucleobases (ie, 8 consecutive nucleobases) of a target region to which an oligomeric active compound is targeted.

[000156] "Wing segment" means a plurality of nucleosides modified to impart to an oligonucleotide properties, such as enhanced inhibitory activity, increased binding affinity for a target nucleic acid, or resistance to degradation by nucleases in vivo.

Certain Modalities

[000157] Certain embodiments provide methods, compounds, and compositions for inhibiting HBV mRNA expression.

[000158] Certain embodiments deliver target antisense compounds to an HBV nucleic acid. In certain embodiments, the HBV nucleic acid is the sequence set forth in GENBANK Accession number U95551.1 (incorporated herein as SEQ ID NO: 1).

[000159] In certain embodiments, the compounds provided herein are or comprise a modified oligonucleotide. In certain embodiments, the compounds comprise a modified oligonucleotide and a conjugate, as described herein. In certain embodiments, the modified oligonucleotide is a pharmaceutically acceptable derivative.

[000160] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide with 10 to 30 linked nucleosides in length targeted to HBV. The HBV target may have a sequence referred to in SEQ ID NO: 1 or a portion thereof, or a variant thereof.

[000161] In certain embodiments, the compounds or modified oligonucleotides provided herein are 10 to 30 linked nucleosides in length and are targeted to HBV. In certain embodiments, the HBV target has the sequence referred to in SEQ ID NO: 1. In certain embodiments, these target compounds or oligonucleotides are one of the following HBV nucleotide regions: CCTGCTGGTGGCCTCCAGTTC (SEQ ID NO: 1273); AGAGTCTAGACTCGTGGTGGACTTCCTTCA (SEQ ID NO: 1354); CATCCTGCTGGCTATGCCTCATCTTCTT (SEQ ID NO: 1276); CAAGGTATGTTGCCCGT (SEQ ID NO: 1277); CCTATGGGAGTGGGCCTCAG (SEQ ID NO: 1279; TGGCTCAGTTTACTAGTGCCATTTGTTCAGTGGTTCG (SEQ ID NO: 1287); TATATGGATGATGATGTGGT (SEQ ID NO:1359); TGCCAAGTGTTTGCTGA (SEQ ID NO:1360); TGCCGATCCATACTGCGGAACTCCT (SEQ ID NO: 1361); C CGTGTGCACTTCGCTTCACCTCTGCACGT (SEQ ID NO:1352); GGAGGCTGTAGGCATAAATTGGT (SEQ ID NO:1353); CTTTTTCACCTCTGCCTA (SEQ ID NO:1362); TTCAAGCCTCCAAGCTGTGCCTTGG (SEQ ID NO:1363); AGAGTCTAGACTCGTGGTGGACTTCCTTCAATTTTCTAGGGG (SEQ ID NO: 1274); TGGATGTGTCTGCGGCGTTTTATCAT (SEQ ID NO: 1275); TGTATTCCCATCCCATC (SEQ ID NO: 1278); TGGCTCAGTTTACTAGTGC (SEQ ID NO: 1280); GGGCTTTCCCCCACTGT (SEQ ID NO: 1281); TCCTCTGCCGATCCATACTGCGGAACTCCT (SEQ ID NO: 1) 282); CGCACCCTCTCTTTACGCGG (SEQ ID NO: 1283); GGAGTGTGGATTCGCAC (SEQ ID NO: 1284); or GAAGAAGAACTCCCTCGCCT (SEQ ID NO: 1285). In certain embodiments, such compounds or oligonucleotides have a gap segment of 9, 10 or more deoxynucleoside bonds. gap of 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 linked nucleosides. In certain embodiments, such a gap segment is between two wing segments that independently have 1, 2, 3, 4, 5, 6, 7 or 8, linked modified nucleosides. In certain embodiments, one or more modified nucleosides in the wing segment have a modified sugar. In certain embodiments, the modified sugar is a bicyclic sugar. In certain embodiments, the modified nucleoside is an LNA nucleoside. In certain embodiments, the modified nucleoside is a 2'-substituted nucleoside. In certain embodiments, the 2'-substituted nucleosides include nucleosides with bicyclic sugar modifications. In certain embodiments, the modified nucleoside is a 2'-MOE nucleoside. In certain embodiments, the modified nucleoside is a restricted ethyl (cEt) nucleoside.

[000162] In certain embodiments, the compounds or compositions comprise modified oligonucleotides consisting of 10 to 30 linked nucleosides and having a sequence of nucleobases comprising a portion with at least 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 contiguous nucleobases complementary to an equal length portion of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 12 84, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1. In certain embodiments, such oligonucleotides have a gap segment of 9, 10, or more deoxynucleosides attached. In certain embodiments, such a gap segment is between two wing segments that independently have 1, 2, 3, 4, 5, 6, 7, or 8 modified nucleosides attached. In certain embodiments, one or more modified nucleosides in the wing segment have a modified sugar. In certain embodiments, the modified sugar is a bicyclic sugar. In certain embodiments, the modified nucleoside is an LNA nucleoside. In certain embodiments, the modified nucleoside is a 2'-substituted nucleoside. In certain embodiments, 2'-substituted nucleosides include nucleosides with bicyclic sugar modifications. In certain embodiments, the modified nucleoside is a 2'-MOE nucleoside. In certain embodiments, the modified nucleoside is a restricted ethyl (cEt) nucleoside. In certain embodiments, each modified nucleoside in each wing segment is independently a 2'-MOE nucleoside or a nucleoside with a bicyclic sugar modification such as a restricted ethyl (cEt) nucleoside or LNA nucleoside.

[000163] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 10 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobases of any one of SEQ ID NOs: 531 0, 321-802, 804-1272, 1288-1350, 1364-1372, 1375, 1376, or 1379. In certain embodiments, such oligonucleotides have a gap segment of 9, 10, or more deoxynucleosides attached. In certain embodiments, such a gap segment is between two wing segments that independently have 1-5, 1-4, 1-3, 2-5, 2-4 or 2-3 modified nucleosides attached. In certain embodiments, such a gap segment is between two wing segments that independently have 1, 2, 3, 4, 5, 6, 7, or 8 modified nucleosides attached. In certain embodiments, one or more modified nucleosides in the wing segment have a modified sugar. In certain embodiments, the modified sugar is a bicyclic sugar. In certain embodiments, the modified nucleoside is an LNA nucleoside. In certain embodiments, the modified nucleoside is a 2'-substituted nucleoside. In certain embodiments, 2'-substituted nucleosides include nucleosides with bicyclic sugar modifications. In certain embodiments, the modified nucleoside is a 2'-MOE nucleoside. In certain embodiments, the modified nucleoside is a restricted ethyl (cEt) nucleoside.

[000164] In certain embodiments, the modified oligonucleotide is 16 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, or 4 modified sugar nucleosides. In certain embodiments, each sugar-modified nucleoside is independently a 2'-MOE nucleoside or a nucleoside with a bicyclic sugar moiety such as a restricted ethyl (cEt) nucleoside or LNA nucleoside. In certain embodiments, the modified oligonucleotide is 16 nucleosides in length and has a gap segment of 9 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, 4, or 5 modified sugar nucleosides. In certain embodiments, each sugar-modified nucleoside is independently a 2'-MOE nucleoside or a bicyclic nucleoside such as a restricted ethyl (cEt) nucleoside or LNA nucleoside.

[000165] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 10 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobases of any one of SEQ ID NOs: 531 0, 321-802, 804-1272, 1288-1350, 1364-1372, 1375, 1376, or 1379. In certain embodiments, such oligonucleotides have a gap segment of 10 or more deoxynucleosides attached. In certain embodiments, such a gap segment is between two wing segments that independently have 1-5, 1-4, 1-3, 2-5, 2-4 or 2-3 modified nucleosides attached. In certain embodiments, such a gap segment is between two wing segments that independently have 2, 3, 4, 5, 6, 7, or 8 modified nucleosides attached. In certain embodiments, one or more modified nucleosides in the wing segment have a modified sugar. In certain embodiments, the modified sugar is a bicyclic sugar. In certain embodiments, the modified nucleoside is an LNA nucleoside. In certain embodiments, the modified nucleoside is a 2'-substituted nucleoside. In certain embodiments, 2'-substituted nucleosides include nucleosides with bicyclic sugar modifications. In certain embodiments, the modified nucleoside is a 2'-MOE nucleoside.

[000166] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 10 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobases of any one of SEQ ID NOs: 5, 15, 16, 33, 39-95, 123-135, 163-175, 180-310, 321-406, 413-455, 461802, or 804-1272. In certain embodiments, such oligonucleotides have a gap segment of 10 or more deoxynucleosides attached. In certain embodiments, such a gap segment is between two wing segments that independently have 1-5, 1-4, 1-3, 2-5, 2-4 or 2-3 modified nucleosides attached. In certain embodiments, one or more modified nucleosides in the wing segment have a modified sugar. In certain embodiments, the modified sugar is a bicyclic sugar. In certain embodiments, the modified nucleoside is an LNA nucleoside. In certain embodiments, the modified nucleoside is a 2'-substituted nucleoside. In certain embodiments, 2'-substituted nucleosides include nucleosides with bicyclic sugar modifications. In certain embodiments, the modified nucleoside is a 2'-MOE nucleoside.

[000167] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 10 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobases of any one of SEQ ID NOs: 6-1 4, 17-32, 34-38, 96-122, 136-162, 176-179, 407-412, 456-462, 523-538. In certain embodiments, such oligonucleotides have a gap segment of 10 or more deoxynucleosides attached. In certain embodiments, such a gap segment is between two wing segments that independently have 1-5, 1-4, 1-3, 2-5, 2-4 or 2-3 modified nucleosides attached. In certain embodiments, one or more modified nucleosides in the wing segment have a modified sugar. In certain embodiments, the modified nucleoside is a 2'-substituted nucleoside. In certain embodiments, the modified nucleoside is a 2'-MOE nucleoside. In certain embodiments, the modified oligonucleotide is 14 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-3 or 2 modified sugar nucleosides.

[000168] In certain embodiments, the modified oligonucleotide is 16 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, 2-4 or 3 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 16 nucleosides in length and has a gap segment of 9 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, 4, or 5 modified sugar nucleosides, such as 2'-MOE nucleosides.

[000169] In certain embodiments, the modified oligonucleotide is 17 nucleosides in length and has a gap segment of 9 or 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, 2-4 or 3-4 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, 4, 5, or 6 modified sugar nucleosides, such as 2'-MOE nucleosides.

[000170] In certain embodiments, the modified oligonucleotide is 18 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, 3-5, or 4 modified sugar nucleosides.

[000171] In certain embodiments, the modified oligonucleotide is 20 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, or 5 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, 4, 5, 6, 7, or 8 modified sugar nucleosides, such as 2'-MOE nucleosides.

[000172] In certain embodiments, the compounds or compositions comprise a salt of the modified oligonucleotide.

[000173] In certain embodiments, the compounds or compositions further comprise a pharmaceutically acceptable carrier or diluent.

[000174] In certain embodiments, the nucleobase sequence of the modified oligonucleotide is at least 70%, 75%, 80%, 85%, 90%, 95% or 100% complementary to SEQ ID NO: 1, as measured over the entire modified oligonucleotide.

[000175] In certain embodiments, the nucleobase sequence of the modified oligonucleotide is at least 70%, 75%, 80%, 85%, 90%, 95% or 100% complementary to any one of SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 1279, 128 0, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, as measured over the entire modified oligonucleotide.

[000176] In certain embodiments, the compound or modified oligonucleotide is single-stranded.

[000177] In certain embodiments, the modified oligonucleotide consists of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 linked nucleosides. In certain embodiments, the modified oligonucleotide consists of 20 linked nucleosides. In certain embodiments, the modified oligonucleotide consists of 18 linked nucleosides. In certain embodiments, the modified oligonucleotide consists of 17 linked nucleosides. In certain embodiments, the modified oligonucleotide consists of 16 linked nucleosides. In certain embodiments, the modified oligonucleotide consists of 14 linked nucleosides.

[000178] In certain embodiments, at least one internucleoside linkage of the modified oligonucleotide is a modified internucleoside linkage. In certain embodiments, each internucleoside linkage is a phosphorothioate internucleoside linkage.

[000179] In certain embodiments, at least one nucleoside of the modified oligonucleotide comprises a modified sugar. In certain embodiments, the at least one modified sugar comprises a 2'-O-methoxyethyl (2'-O(CH2)2-OCH3) group. In certain embodiments, the modified sugar comprises a 2'-O-CH3 group.

[000180] In certain embodiments, at least one modified sugar is a bicyclic sugar. In certain embodiments, at least one modified sugar, the bicyclic sugar, comprises a 4'-(CH 2 -O-2' bridge, where n is 1 or 2. In certain embodiments, the bicyclic sugar comprises a 4'-CH 2 -O-2' bridge. In certain embodiments, the bicyclic sugar comprises a 4'-CH(CH3 )-O-2' bridge.

[000181] In certain embodiments, at least one nucleoside of said modified oligonucleotide comprises a modified nucleobase. In certain embodiments, the modified nucleobase is a 5-methylcytosine.

[000182] In certain embodiments, the modified oligonucleotide consists of a single-stranded modified oligonucleotide.

[000183] In certain embodiments, the modified oligonucleotide comprises: a) a gap segment consisting of linked deoxynucleosides; b) a 5' wing segment consisting of linked nucleosides; and c) a 3' wing segment consisting of linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment and each nucleoside of each wing segment comprises a modified sugar.

[000184] In certain embodiments, the modified oligonucleotide consists of 16 linked nucleosides, the gap segment consisting of 10 linked deoxynucleosides, the 5' wing segment consisting of three linked nucleosides, the 3' wing segment consisting of three linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar and/or an ethyl sugar (cEt) restrict do, each internucleoside linkage is a phosphorothioate linkage and each cytosine is a 5-methylcytosine. In some aspects, each of the three linked nucleosides of the 5' wing segment is a 2'-O-methoxyethyl sugar and each of the three linked nucleosides of the 3' wing segment is a restricted ethyl (cEt) sugar. In other aspects, the three linked nucleosides of the 5'-wing segment are a 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, and an ethyl (cEt) sugar restricted in the 5' to 3' direction, and the three linked nucleosides of the 3' wing segment are a restricted ethyl (cEt) sugar, a restricted ethyl (cEt) sugar, and a 2'-O sugar - methoxyethyl in the 5' to 3' direction. In other aspects, the three linked nucleosides of the 5' wing segment are a restricted ethyl (cEt) sugar, a 2'-O-methoxyethyl sugar, and an ethyl (cEt) sugar restricted in the 5' to 3' direction, and the three linked nucleosides of the 3' wing segment are a restricted ethyl (cEt) sugar, a 2'-O-methoxyethyl sugar, and an ethyl sugar (c Et) constrained in the 5' to 3' direction.

[000185] In certain embodiments, the modified oligonucleotide consists of 16 linked nucleosides, the gap segment consisting of 10 linked deoxynucleosides, the 5' wing segment consisting of two linked nucleosides, the 3' wing segment consisting of four linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar and a restricted ethyl (cEt) sugar, each internucleoside linkage is a phosphorothioate linkage and each cytosine is a 5-methylcytosine. In some aspects, the two linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar and an ethyl (cEt) sugar restricted in the 5' to 3' direction, and the four linked nucleosides of the 3' wing segment are a restricted ethyl (cEt) sugar, a 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, and a 2'-O sugar -methoxyethyl in the 5' to 3' direction.

[000186] In certain embodiments, the modified oligonucleotide consists of 16 linked nucleosides, the gap segment consisting of 9 linked deoxynucleosides, the 5' wing segment consisting of five linked nucleosides, the 3' wing segment consisting of two linked nucleosides; the five linked nucleosides of the 5' wing segment are a restricted ethyl (cEt) sugar, a 2'-deoxynucleoside, a restricted ethyl (cEt) sugar, a 2'-deoxynucleoside, and a 5' to 3' restricted ethyl (cEt) sugar; the two linked nucleosides of the 3' wing segment are a 2'-O-methoxyethyl sugar and a 2'-O-methoxyethyl sugar in the 5' to 3' direction; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000187] In certain embodiments, the modified oligonucleotide consists of 16 linked nucleosides, the gap segment consisting of 8 linked deoxynucleosides, the 5' wing segment consisting of three linked nucleosides, the 3' wing segment consisting of five linked nucleosides; the three linked nucleosides of the 5' wing segment are an ethyl (cEt) sugar restricted, a 2'-O-methoxyethyl sugar, and an ethyl (cEt) sugar restricted in the 5' to 3' direction; each of the five linked nucleosides of the 3' wing segment is a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000188] In certain embodiments, the modified oligonucleotide consists of 16 linked nucleosides, the gap segment consisting of 8 linked deoxynucleosides, the 5' wing segment consisting of three linked nucleosides, the 3' wing segment consisting of five linked nucleosides; each of the three linked nucleosides of the 5' wing segment is a restricted ethyl (cEt) sugar; each of the five linked nucleosides of the 3' wing segment is a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000189] In certain embodiments, the modified oligonucleotide consists of 17 linked nucleosides, the gap segment consisting of 9 linked deoxynucleosides, the 5' wing segment consisting of four linked nucleosides, the 3' wing segment consisting of four linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar and a restricted ethyl sugar (cEt), each internucleoside linkage is a phosphorothioate linkage and each cytosine is a 5-methylcytosine. In some aspects, the four linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, and an ethyl (cEt) sugar restricted in the 5' to 3' direction, and the four linked nucleosides of the 3' wing segment are a 2'-O-methoxyethyl sugar, ethyl sugar (c Et) restricted, a 2'-O-methoxyethyl sugar, and a 2'-O-methoxyethyl sugar in the 5' to 3' direction.

[000190] In certain embodiments, the modified oligonucleotide consists of 17 linked nucleosides, the gap segment consisting of 9 linked deoxynucleosides, the 5' wing segment consisting of four linked nucleosides, the 3' wing segment consisting of four linked nucleosides; the four linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, and an ethyl (cEt) sugar restricted in the 5' to 3' direction; the four linked nucleosides of the 3' wing segment are a restricted ethyl (cEt) sugar, a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, and a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000191] In certain embodiments, the modified oligonucleotide consists of 17 linked nucleosides, the gap segment consisting of 9 linked deoxynucleosides, the 5' wing segment consisting of four linked nucleosides, the 3' wing segment consisting of four linked nucleosides; the four linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, a 2'-O-methoxyethyl sugar, and an ethyl (cEt) sugar restricted in the 5' to 3' direction; each of the four linked nucleosides of the 3' wing segment is a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000192] In certain embodiments, the modified oligonucleotide consists of 17 linked nucleosides, the gap segment consisting of 8 linked deoxynucleosides, the 5' wing segment consisting of five linked nucleosides, the 3' wing segment consisting of four linked nucleosides; the five linked nucleosides of the 5' wing segment are a restricted ethyl (cEt) sugar, a 2'-deoxynucleoside, a restricted ethyl (cEt) sugar, a 2'-deoxynucleoside, and a 5' to 3' restricted ethyl (cEt) sugar; each of the four linked nucleosides of the 3' wing segment is a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000193] In certain embodiments, the modified oligonucleotide consists of 17 linked nucleosides, the gap segment consisting of 8 linked deoxynucleosides, the 5' wing segment consisting of five linked nucleosides, the 3' wing segment consisting of four linked nucleosides; the five linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, and a 5' to 3' restricted ethyl (cEt) sugar; each of the four linked nucleosides of the 3' wing segment is a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000194] In certain embodiments, the modified oligonucleotide consists of 17 linked nucleosides, the gap segment consisting of 7 linked deoxynucleosides, the 5' wing segment consisting of five linked nucleosides, the 3' wing segment consisting of five linked nucleosides; the five linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, and a 5' to 3' restricted ethyl (cEt) sugar; the five linked nucleosides of the 3' wing segment are a restricted ethyl (cEt) sugar, a restricted ethyl (cEt) sugar, a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, and a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000195] In certain embodiments, the modified oligonucleotide consists of 17 linked nucleosides, the gap segment consisting of 7 linked deoxynucleosides, the 5' wing segment consisting of six linked nucleosides, the 3' wing segment consisting of four linked nucleosides; the six linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, and a 5' to 3' restricted ethyl (cEt) sugar; each of the four linked nucleosides of the 3' wing segment is a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000196] In certain embodiments, the modified oligonucleotide consists of 17 linked nucleosides, the gap segment consisting of 7 linked deoxynucleosides, the 5' wing segment consisting of six linked nucleosides, the 3' wing segment consisting of four linked nucleosides; the six linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, a 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, a 2'-deoxynucleoside, and a 5' to 3' restricted ethyl (cEt) sugar; each of the four linked nucleosides of the 3' wing segment is a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000197] In certain embodiments, the modified oligonucleotide consists of 20 linked nucleosides, the gap segment consisting of 10 linked deoxynucleosides, the 5' wing segment consisting of five linked nucleosides, the 3' wing segment consisting of five linked nucleosides, each nucleoside of each wing segment comprises a restricted ethyl sugar (cEt), each internucleoside linkage is a phosphorothioate linkage and each cytosine is a 5-methylcytosine.

[000198] In certain embodiments, the modified oligonucleotide consists of 20 linked nucleosides, the gap segment consisting of 10 linked deoxynucleosides, the 5' wing segment consisting of five linked nucleosides, the 3' wing segment consisting of five linked nucleosides, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine, in which the five linked nucleosides of the 5' wing are a restricted ethyl (cEt) sugar, a 2'-deoxynucleoside, a restricted ethyl (cEt) sugar, a 2'-deoxynucleoside, and an ethyl (cEt) sugar restricted in the 5' to 3' direction, and each of the five linked nucleosides of the 3' wing is a 2'-O-methoxyethyl sugar.

[000199] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 16 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 12 79, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of three linked nucleosides; and c) a 3' wing segment consisting of three linked nucleosides. In some aspects, the gap segment is positioned between the 5' wing segment and the 3' wing segment; each of the three linked nucleosides of the 5' wing segment is a 2'-O-methoxyethyl sugar and each of the three linked nucleosides of the 3' wing segment is a restricted ethyl (cEt) sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine residue is a 5-methylcytosine. In other respects, the gap segment is positioned between the 5' wing segment and the 3' wing segment; the three linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, and an ethyl (cEt) sugar restricted in the 5' to 3' direction; the three linked nucleosides of the 3' wing segment are a restricted ethyl (cEt) sugar, a restricted ethyl (cEt) sugar, and a 2'-O-methoxyethyl sugar in the 5' to 3' direction; each internucleoside linkage is a phosphorothioate linkage; and each cytosine residue is a 5-methylcytosine. In other aspects, the three linked nucleosides of the 5' wing segment are a restricted ethyl (cEt) sugar, a 2'-O-methoxyethyl sugar, and an ethyl (cEt) sugar restricted in the 5' to 3' direction, and the three linked nucleosides of the 3' wing segment are a restricted ethyl (cEt) sugar, a 2'-O-methoxyethyl sugar, and an ethyl sugar (c Et) constrained in the 5' to 3' direction.

[000200] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 16 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 12 79, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises the ) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of two linked nucleosides; and c) a 3' wing segment consisting of four linked nucleosides. In some aspects, the gap segment is positioned between the 5' wing segment and the 3' wing segment; the two linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar and an ethyl (cEt) sugar restricted in the 5' to 3' direction; the four linked nucleosides of the 3' wing segment are a restricted ethyl (cEt) sugar, 2'-O-methoxyethyl sugar, restricted ethyl (cEt) sugar, and 2'-O-methoxyethyl sugar in the 5' to 3' direction; each internucleoside linkage is a phosphorothioate linkage; and each cytosine residue is a 5-methylcytosine.

[000201] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 16 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 12 79, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of 9 linked deoxynucleosides; b) a 5' wing segment consisting of five linked nucleosides; and c) a 3' wing segment consisting of two linked nucleosides, wherein the five linked nucleosides of the 5' wing segment are a restricted ethyl (cEt) sugar, a 2'-deoxynucleoside, a restricted ethyl (cEt) sugar, a 2'-deoxynucleoside, and a 5' to 3' restricted ethyl (cEt) sugar; the two linked nucleosides of the 3' wing segment are a 2'-O-methoxyethyl sugar and a 2'-O-methoxyethyl sugar in the 5' to 3' direction; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000202] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 16 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 12 79, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of 8 linked deoxynucleosides; b) a 5' wing segment consisting of three linked nucleosides; and c) a 3' wing segment consisting of five linked nucleosides, wherein the three linked nucleosides of the 5' wing segment are an ethyl (cEt) sugar restricted, a 2'-O-methoxyethyl sugar, and an ethyl (cEt) sugar restricted in the 5' to 3' direction; each of the five linked nucleosides of the 3' wing segment is a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000203] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 16 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 12 79, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of 8 linked deoxynucleosides; b) a 5' wing segment consisting of three linked nucleosides; and c) a 3' wing segment consisting of five linked nucleosides, wherein each of the three linked nucleosides of the 5' wing segment is a restricted ethyl (cEt) sugar; each of the five linked nucleosides of the 3' wing segment is a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000204] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 17 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 12 79, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of 9 linked deoxynucleosides; b) a 5' wing segment consisting of four linked nucleosides; and c) a 3' wing segment consisting of four linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar and a restricted ethyl (cEt) sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine. In some aspects, the four linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, and an ethyl (cEt) sugar restricted in the 5' to 3' direction, and the four linked nucleosides of the 3' wing segment are a 2'-O-methoxyethyl sugar, ethyl sugar (c Et) restricted, a 2'-O-methoxyethyl sugar, and a 2'-O-methoxyethyl sugar in the 5' to 3' direction.

[000205] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 17 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 12 79, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of 9 linked deoxynucleosides; b) a 5' wing segment consisting of four linked nucleosides; and c) a 3' wing segment consisting of four linked nucleosides, wherein the four linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, and an ethyl (cEt) sugar restricted in the 5' to 3' direction; the four linked nucleosides of the 3' wing segment are a restricted ethyl (cEt) sugar, a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, and a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000206] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 17 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 12 79, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of 9 linked deoxynucleosides; b) a 5' wing segment consisting of four linked nucleosides; and c) the 3' wing segment consisting of four linked nucleosides, wherein the four linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, a 2'-O-methoxyethyl sugar, and a 5' to 3' restricted ethyl (cEt) sugar; each of the four linked nucleosides of the 3' wing segment is a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000207] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 17 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 12 79, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of 8 linked deoxynucleosides; b) a 5' wing segment consisting of five linked nucleosides; and c) a 3' wing segment consisting of four linked nucleosides, wherein the five linked nucleosides of the 5' wing segment are a restricted ethyl (cEt) sugar, a 2'-deoxynucleoside, a restricted ethyl (cEt) sugar, a 2'-deoxynucleoside, and a 5' to 3' restricted ethyl (cEt) sugar; each of the four linked nucleosides of the 3' wing segment is a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000208] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 17 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 12 79, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of 8 linked deoxynucleosides; b) a 5' wing segment consisting of five linked nucleosides; and c) a 3' wing segment consisting of four linked nucleosides, wherein the five linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, and a 5' to 3' restricted ethyl (cEt) sugar; each of the four linked nucleosides of the 3' wing segment is a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000209] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 17 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 12 79, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of 7 linked deoxynucleosides; b) a 5' wing segment consisting of five linked nucleosides; and c) a 3' wing segment consisting of five linked nucleosides, wherein the five linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, and a 5' to 3' restricted ethyl (cEt) sugar; the five linked nucleosides of the 3' wing segment are a restricted ethyl (cEt) sugar, a restricted ethyl (cEt) sugar, a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, and a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000210] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 17 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 12 79, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of 7 linked deoxynucleosides; b) a 5' wing segment consisting of six linked nucleosides; and c) a 3' wing segment consisting of four linked nucleosides, wherein the six linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, a 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, and a restricted ethyl (cEt) sugar in the direction 5' to 3'; each of the four linked nucleosides of the 3' wing segment is a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000211] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 17 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 12 79, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of 7 linked deoxynucleosides; b) a 5' wing segment consisting of six linked nucleosides; and c) a 3' wing segment consisting of four linked nucleosides, wherein the six linked nucleosides of the 5' wing segment are a 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, a 2'-O-methoxyethyl sugar, a restricted ethyl (cEt) sugar, a 2'-deoxynucleoside, and a 5'-restricted ethyl (cEt) sugar to 3'; each of the four linked nucleosides of the 3' wing segment is a 2'-O-methoxyethyl sugar; each internucleoside linkage is a phosphorothioate linkage; and each cytosine is a 5-methylcytosine.

[000212] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 20 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 12 79, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of 10 linked deoxynucleosides; b) a 5' wing segment consisting of five linked nucleosides; and c) a 3' wing segment consisting of five linked nucleosides, each nucleoside of each wing segment comprises a restricted ethyl (cEt) sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000213] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 20 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1, 1273, 1274, 1275, 1276, 1277, 1278, 12 79, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of 10 linked deoxynucleosides; b) a 5' wing segment consisting of five linked nucleosides; and c) a 3' wing segment consisting of five linked nucleosides, each internucleoside linkage is a phosphorothioate linkage and each cytosine is a 5-methylcytosine, wherein the five linked nucleosides of the 5' wing are a restricted ethyl (cEt) sugar, a 2'-deoxynucleoside, a restricted ethyl (cEt) sugar, a 2'-deoxynucleoside, and an ethyl (c) sugar Et) restricted in the 5' to 3' direction, and each of the five linked nucleosides of the 3' wing is a 2'-O-methoxyethyl sugar.

[000214] In certain embodiments, the modified oligonucleotide consists of 20 linked nucleosides, the gap segment consisting of ten linked deoxynucleosides, the 5' wing segment consisting of five linked nucleosides, the 3' wing segment consisting of five linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000215] In certain embodiments, the modified oligonucleotide consists of 20 linked nucleosides, the gap segment consisting of ten linked deoxynucleosides, the 5' wing segment consisting of two linked nucleosides, the 3' wing segment consisting of eight linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000216] In certain embodiments, the modified oligonucleotide consists of 20 linked nucleosides, the gap segment consisting of ten linked deoxynucleosides, the 5' wing segment consisting of eight linked nucleosides, the 3' wing segment consisting of two linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000217] In certain embodiments, the modified oligonucleotide consists of 20 linked nucleosides, the gap segment consisting of ten linked deoxynucleosides, the 5' wing segment consisting of three linked nucleosides, the 3' wing segment consisting of seven linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000218] In certain embodiments, the modified oligonucleotide consists of 20 linked nucleosides, the gap segment consisting of ten linked deoxynucleosides, the 5' wing segment consisting of seven linked nucleosides, the 3' wing segment consisting of three linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000219] In certain embodiments, the modified oligonucleotide consists of 20 linked nucleosides, the gap segment consisting of ten linked deoxynucleosides, the 5' wing segment consisting of four linked nucleosides, the 3' wing segment consisting of six linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000220] In certain embodiments, the modified oligonucleotide consists of 20 linked nucleosides, the gap segment consisting of ten linked deoxynucleosides, the 5' wing segment consisting of six linked nucleosides, the 3' wing segment consisting of four linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000221] In certain embodiments, the modified oligonucleotide consists of 18 linked nucleosides, the gap segment consisting of ten linked deoxynucleosides, the 5' wing segment consisting of four linked nucleosides, the 3' wing segment consisting of four linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000222] In certain embodiments, the modified oligonucleotide consists of 17 linked nucleosides, the gap segment consisting of ten linked deoxynucleosides, the 5' wing segment consisting of three linked nucleosides, the 3' wing segment consisting of four linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000223] In certain embodiments, the modified oligonucleotide consists of 17 linked nucleosides, the gap segment consisting of nine linked deoxynucleosides, the 5' wing segment consisting of two linked nucleosides, the 3' wing segment consisting of six linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000224] In certain embodiments, the modified oligonucleotide consists of 17 linked nucleosides, the gap segment consisting of nine linked deoxynucleosides, the 5' wing segment consisting of six linked nucleosides, the 3' wing segment consisting of two linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000225] In certain embodiments, the modified oligonucleotide consists of 17 linked nucleosides, the gap segment consisting of nine linked deoxynucleosides, the 5' wing segment consisting of four linked nucleosides, the 3' wing segment consisting of four linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000226] In certain embodiments, the modified oligonucleotide consists of 17 linked nucleosides, the gap segment consisting of nine linked deoxynucleosides, the 5' wing segment consisting of five linked nucleosides, the 3' wing segment consisting of three linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000227] In certain embodiments, the modified oligonucleotide consists of 17 linked nucleosides, the gap segment consisting of nine linked deoxynucleosides, the 5' wing segment consisting of three linked nucleosides, the 3' wing segment consisting of five linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000228] In certain embodiments, the modified oligonucleotide consists of 16 linked nucleosides, the gap segment consisting of ten linked deoxynucleosides, the 5' wing segment consisting of three linked nucleosides, the 3' wing segment consisting of three linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000229] In certain embodiments, the modified oligonucleotide consists of 16 linked nucleosides, the gap segment consisting of nine linked deoxynucleosides, the 5' wing segment consisting of five linked nucleosides, the 3' wing segment consisting of two linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000230] In certain embodiments, the modified oligonucleotide consists of 16 linked nucleosides, the gap segment consisting of nine linked deoxynucleosides, the 5' wing segment consisting of two linked nucleosides, the 3' wing segment consisting of five linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000231] In certain embodiments, the modified oligonucleotide consists of 16 linked nucleosides, the gap segment consisting of nine linked deoxynucleosides, the 5' wing segment consisting of four linked nucleosides, the 3' wing segment consisting of three linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000232] In certain embodiments, the modified oligonucleotide consists of 16 linked nucleosides, the gap segment consisting of nine linked deoxynucleosides, the 5' wing segment consisting of three linked nucleosides, the 3' wing segment consisting of four linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000233] In certain embodiments, the modified oligonucleotide consists of 14 linked nucleosides, the gap segment consisting of ten linked deoxynucleosides, the 5' wing segment consisting of two linked nucleosides, the 3' wing segment consisting of two linked nucleosides, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside linkage is a phosphorothioate linkage, and each cytosine is a 5-methylcytosine.

[000234] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 20 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279 , 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of five linked nucleosides; and c) a 3' wing segment consisting of five linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000235] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 20 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of two linked nucleosides; and c) a 3' wing segment consisting of eight linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000236] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 20 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of eight linked nucleosides; and c) a 3' wing segment consisting of two linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000237] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 20 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of three linked nucleosides; and c) a 3' wing segment consisting of seven linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000238] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 20 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of seven linked nucleosides; and c) a 3' wing segment consisting of three linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000239] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 20 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of four linked nucleosides; and c) a 3' wing segment consisting of six linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000240] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 20 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of six linked nucleosides; and c) a 3' wing segment consisting of four linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000241] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 18 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of four linked nucleosides; and c) a 3' wing segment consisting of four linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000242] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 17 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of three linked nucleosides; and c) a 3' wing segment consisting of four linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000243] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 17 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of nine linked deoxynucleosides; b) a 5' wing segment consisting of two linked nucleosides; and c) a 3' wing segment consisting of six linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000244] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 17 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of nine linked deoxynucleosides; b) a 5' wing segment consisting of six linked nucleosides; and c) a 3' wing segment consisting of two linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000245] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 17 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of nine linked deoxynucleosides; b) a 5' wing segment consisting of four linked nucleosides; and c) a 3' wing segment consisting of four linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000246] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 17 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of nine linked deoxynucleosides; b) a 5' wing segment consisting of five linked nucleosides; and c) a 3' wing segment consisting of three linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000247] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 17 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of nine linked deoxynucleosides; b) a 5' wing segment consisting of three linked nucleosides; and c) a 3' wing segment consisting of five linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000248] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 16 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of three linked nucleosides; and c) a 3' wing segment consisting of three linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000249] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 16 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of nine linked deoxynucleosides; b) a 5' wing segment consisting of five linked nucleosides; and c) a 3' wing segment consisting of two linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000250] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 16 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of nine linked deoxynucleosides; b) a 5' wing segment consisting of two linked nucleosides; and c) a 3' wing segment consisting of five linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000251] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 16 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of nine linked deoxynucleosides; b) a 5' wing segment consisting of four linked nucleosides; and c) a 3' wing segment consisting of three linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000252] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 16 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of nine linked deoxynucleosides; b) a 5' wing segment consisting of three linked nucleosides; and c) a 3' wing segment consisting of four linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000253] In certain embodiments, the compounds or compositions comprise a modified oligonucleotide consisting of 14 linked nucleosides having a sequence of nucleobases comprising a portion of at least 8 contiguous nucleobases complementary to a portion of equal length of any of the nucleobases set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1354, 1359, 1360, 1361, 1362, and 1363, wherein the nucleobase sequence is complementary to SEQ ID NO: 1 and wherein the modified oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of two linked nucleosides; and c) a 3' wing segment consisting of two linked nucleosides. The gap segment is positioned between the 5' wing segment and the 3' wing segment, each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine residue is a 5-methylcytosine.

[000254] In certain embodiments, the methods, compounds, and compositions provided inhibit HBV mRNA expression and/or DNA levels and/or protein levels and/or antigen levels.

[000255] Another embodiment provides a method for treating diseases, disorders and conditions related to HBV in a mammal, the method comprising administering a therapeutically effective amount of any pharmaceutical composition as described above to a mammal in need thereof, in order to treat the diseases, disorders and conditions related to HBV. In related embodiments, the mammal is a human and the HBV-related disease, disorder and condition is hepatitis B virus infection from a human hepatitis B virus. More particularly, the human hepatitis B virus can be any of the human geographic genotypes: A (Northwestern Europe, North America, Central America); B (Indonesia, China, Vietnam); C (East Asia, Korea, China, Japan, Polynesia, Vietnam); D (Mediterranean Area, Middle East, India); E (Africa); F (Native Americas, Polynesia); G (United States, France); or H (Central America).

[000256] In certain embodiments, an antisense compound or target oligonucleotide for an HBV nucleic acid is complementary within the following nucleotide regions of SEQ ID NO: 1: 1-20, 10-29, 10-56, 13-38, 13-35, 19-38, 25-47, 25-50, 25-56, 43-68, 4 3-63, 55-74, 58-73, 58-74, 58-77, 58-79, 58-80, 58-84, 59-74, 59-75, 59-80, 60-75, 60-76, 60-79, 61-76, 61-77, 61-80, 62-77 . 196-218, 199-228, 199-218, 199224, 200-224, 205-224, 206-228, 218-237, 224-243, 233-264, 242-263, 243-262, 244-263, 245-27 4; 245-260, 245-264, 246-266, 247-266, 247269, 247-270, 245-267, 251-267, 245-266, 250-269, 251-266, 251-268, 251-269, 245-2 69, 245-266, 245-261, 250-265, 250-266, 250-267, 250268, 250-269, 251-270, 252-267, 253-268, 253-269, 253-272, 253-274, 254 -269, 254-270, 254-274, 255-270, 255-271, 255-274, 255-401, 255400, 255-274, 255-273, 255-272, 255-271, 256-271, 256-275, 2 55-276, 256-272, 256-276, 253-275, 256-279, 257-276, 258-273, 259-274, 260279, 262-281, 262-321, 262-315, 262-312, 265-312, 266-288, 266-291, 266-285, 281-321, 281-303, 290-321, 290-312, 292-311, 290-312, 293312, 293-315, 293-321, 296-321, 302-32 1, 324-343, 339-361, 339-367, 348-367, 342-367, 358-392, 358-378, 360-392, 360-383, 360-388, 360385, 362-381, 366-388, 369- 41 1-426, 411-427, 411-430, 411-431, 411-437, 412-431, 411-426, 411-427, 412-428, 412-431, 412-427, 413-433, 413-432, 413-428 ,413-429,413-432,413-433,414-427,415-427,414-429,414-430,414-433,415-428,415-429,415-430,415-431,415-434,416- 431, 416432, 416-429, 416-435, 417-432, 417-433, 417-436, 418-433, 418-435, 418-434, 418-437, 419-435, 419-434, 420-435, 41 9-432, 419-434, 421-436, 422-437, 422-441, 423-436, 425-465, 454-473, 454-472, 457-476, 457-472, 457-473, 454-476, 455-472, 457-485, 458-485, 458-483, 458477, 458-473, 459-485, 460-485, 463-498, 463-485, 466-485, 463-482, 457-491, 458-491, 459-491 460-491 463-491 466-491 472-491 472493 473-492 475-491 459-494 460-494 463-494 466-494 467-498 472-494 475-4 94, 457-473, 457-472, 458-494, 454-494, 457-494, 457473, 485-513, 470-493, 476-519, 485-519, 500-519, 512-534, 512-550, 524 -546, 536-559, 548-567, 548-570, 550-570, 548-594, 554-573, 548576, 560-594, 584-606, 611-645, 617-363, 623-642, 617-645, 6 39-754, 639-658, 639-654, 641-656, 642-657, 643-658, 642-754, 653-672, 662 685, 665-685,665-689, 668-687, 670-754, 670-706, 670-685, 670-686, 670-689, 671-690, 671-691, 671-686, 671-687, 672-693, 672-697, 672707, 672-687, 672-688, 673-688, 674-69 3, 678-693, 679-694, 679-707, 679-698, 679-701, 679-702, 679-707, 680-695, 680-699, 679-699, 681706, 681-696, 682-697, 682- 706, 682-707, 682-702, 682-701, 683-698, 684-699, 685-700, 686-701, 687-754, 688-704, 689-709, 689-710, 690705, 679-705, 67 9-710, 679-706, 690-710, 691-710, 690-754, 690-706, 684-703, 687-705, 687-702, 687-703, 687-706, 688-703, 688-704, 688705, 689-704, 689-705, 689-708, 690-705, 690-706, 690-709, 691-706, 692-711, 693-716, 693-712, 695-715, 697-716, 697-716, 690-71 6, 724746, 724-752, 724-754, 724-758, 733-752, 738-754, 738-753, 739-758, 739-754, 739-775, 739-754, 740-754, 742-785, 742- 773, 757-776, 757785, 790-815, 793-812, 811-833, 811-844, 814-833, 811-906, 820-839, 822-844, 822-867, 823-842, 845-864, 84 5-867, 854-906, 845-909, 845906, 854-876, 863-882, 863-885, 878-900, 887-906, 899-918, 899-933, 899-958, 905-927, 905-933, 914-933, 936-958, 936-955, 945-964, 951-970, 951-985, 951-1044, 951-1024, 951-1056, 951-997, 960-985, 9631044, 963-1024, 9 63-997, 972-1015, 1025-1044, 1031-1056, 10371056, 1046-1083, 1049-1068, 1070-1089, 1070-1095, 1082-1101, 1081-1134, 1081-11 43, 1082-1101, 1088-1107, 1088-1134, 10941119, 1097-1119, 1112-1134, 1118-1143, 1118-1146, 1088-1146, 1121-1140, 1127-1146, 1127-1193, 1150-1193, 1156-1187, 11651187, 1170-1192, 1171-1191, 1172-1191, 1176-1192, 1176-1285, 1177-1192, 1176-1191, 12 03-1297, 1206-1228, 1206-1255, 1209-1228, 1215-1255, 1245-1265, 1251-1280, 1262-1285, 1251-1285, 1259-1296, 1259-1290, 1259- 1287, 1261-1296, 1261-1285, 1261-1276, 1261-1277, 1261-1280, 1262-1296, 1262-1277, 1262-1278, 1262-1281, 1263-1278, 1263-127 9, 1263-1282, 1264-1279, 1264-1280, 1264-1283, 1264-1297, 1265-1280, 1265-1281, 1265-1284, 1266-1281, 1266-1282, 1266-1285, 1267-1282, 1267-1283, 1268-1283, 1268-1284, 1268-1296, 1269-1284, 1269-1285, 1269-1288, 1270-1285, 1271-1290, 1271-1296, 127 7-1296, 1261-1290, 12621290, 1268-1290, 1263-1305, 1259-1305, 1259-1305, 1266-1305, 1259-1302, 1275-1294, 1281-1306, 1281-1 324, 1281-1336, 12821301, 1286-1306, 1290-1324, 1293-1318, 1290-1324, 1293-1315, 1296-1315, 1311-1336, 1311-1333, 1326-1345 . 515-1602, 1515-1540, 1515-1535, 1518-1605, 1518-1602, 1518-1537, 1521-1563, 1521-1540, 1550-1655, 1550-1563, 1550-1569, 1553 -1578, 1553-1599, 1553-1590, 1565-1584, 1571-1595, 1577-1605, 1577-1606, 1577-1592, 1577-1593, 1577-1596, 1578-1593, 1578-15 94, 1578-1597, 1578-1598, 1571-1598, 1579-1594, 1579-1594, 1579-1598, 1580-1595, 1580-1596, 1580-1599, 1580-1605, 1580-1602, 15811596, 1581-1597, 1581-1600, 1582-1597, 1582-1598, 1582-1601, 1582-1602, 1553-1655, 1583-1598, 1583-1599, 1583-1602, 15 84-1599, 1584-1600, 1584-1603, 1585-1600, 1585-1601, 1585-1604, 1586-1601, 1586-1602, 1586-1605, 1587-1602, 1587-1603, 158 71606, 1588-1603, 1588-1604, 1589-1604, 1589-1605, 1590-1605, 1590-1606, 1591-1606, 1586-1652, 1642-1664, 1651-1720, 165116 73, 1655-1679, 1695-1720, 1716-1738, 1743-1763, 1743-1768, 1764-1783, 1773-1792, 1777-1796, 1777-1800, 1778-1800, 1778-1793, 1778-1794, 1778-1797, 1779-1798, 1779-1797, 1779-1796, 1779-1795, 1779-1794, 1780-1799, 1780-1796, 1780-1795, 17811797, 17 81-1796, 1781-1796, 1781-1800, 1781-1797, 1782-1799, 1782-1797, 1782-1798, 1783-1798, 1783-1799, 1784-1800, 1784-1799, 1779- 1799, 1778-1889, 1778-1794, 1779-1795, 1780-1799, 1785-1800, 1794-1813, 1806-1837, 1806-1828, 1806-1825, 18091828, 1812-184 3, 1812-1837, 1812-1831, 1815-1843, 1815-1844, 1815-1840, 1815-1834, 1818-1837, 1821-1840, 1821-1844, 1821-1837, 1822-1843, 1822-1839, 1822-1837, 1823-1843, 1823-1838, 1824-1839, 1827-1846, 1861-1884, 1861-1880, 1865-1885, 1866-1881, 1867-1882, 186 7-1886, 1868-1883, 1869-1885, 1869-1884, 1870-1885, 1871-1886, 1872-1887, 1874-1889, 1876-1895, 18881914, 1888-1908, 1891-1 910, 1891-1914, 1895-1938, 1895-1935, 1913-1935, 1898-1920, 1907-1929, 1913-1935, 1918-1934, 1919-1938, 1919-1934, 1921-1934 2 368-2396, 2420-2439, 2458-2476, 2459-2478, 2819-2838, 2818-2838, 2873-2892, and 3161-3182.

[000257] In certain embodiments, an antisense compound or oligonucleotide targeting an HBV nucleic acid targets the following nucleotide regions of SEQ ID NO: 1: 1-20, 10-29, 1056, 13-38, 13-35, 19-38, 25-47, 25-50, 25-56, 43-68, 43-6 3, 55-74, 5873, 58-74, 58-77, 58-79, 58-80, 58-84, 59-74, 59-75, 59-80, 60-75, 6076, 60-79, 61-76, 61-77, 61-80, 62-77, 63- 196 -218, 199-228, 199-218, 199-224, 200224, 205-224, 206-228, 218-237, 224-243, 233-264, 242-263, 243-262, 244-263, 245-274; 245-260, 245-264, 246-266, 247-266, 247-269, 247270, 245-267, 251-267, 245-266, 250-269, 251-266, 251-268, 251-269, 245-2 69, 245-266, 245-261, 250-265, 250-266, 250-267, 250-268, 250269, 251-270, 252-267, 253-268, 253-269, 253-272, 253-274, 254 -269, 254-270, 254-274, 255-270, 255-271, 255-274, 255-401, 255-400, 255274, 255-273, 255-272, 255-271, 256-271, 256-275, 2 55-276, 256-272, 256-276, 253-275, 256-279, 257-276, 258-273, 259-274, 260-279, 262281, 262-321, 262-315, 262-312, 265-312, 266-288, 266-291, 266-285, 281-321, 281-303, 290-321, 290-312, 292-311, 290-312, 293-312, 293315, 293-321, 296-321, 302-32 1, 324-343, 339-361, 339-367, 348-367, 342-367, 358-392, 358-378, 360-392, 360-383, 360-388, 360-385, 362381, 366-388, 369- 41 1-426, 411-427, 411-430, 411-431, 411-437, 412-431, 411-426, 411-427, 412-428, 412-431, 412-427, 413-433, 413-432, 413-428, 413429, 413-432, 413-433, 414-427, 415-427, 414-429, 414-430, 414-433, 415-428, 415-429, 415-430, 415-431, 415-434, 416-43 1, 416-432, 416-429, 416-435, 417-432, 417-433, 417-436, 418-433, 418-435, 418-434, 418-437, 419-435, 419-434, 420-435, 419- 432, 419-434, 421-436, 422437, 422-441, 423-436, 425-465, 454-473, 454-472, 457-476, 457-472, 457-473, 454-476, 455-472, 45 7-485, 458-485, 458-483, 458-477, 458473, 459-485, 460-485, 463-498, 463-485, 466-485, 463-482, 457-491, 458-491, 459-491, 460-491, 463-491, 466-491, 472-491, 472-493, 473492, 475-491, 459-494, 460-494, 463-494, 466-494, 467-498, 472-494, 475-494 , 457-473, 457-472, 458-494, 454-494, 457-494, 457-473, 485513, 470-493, 476-519, 485-519, 500-519, 512-534, 512-550, 524-5 46, 536-559, 548-567, 548-570, 550-570, 548-594, 554-573, 548-576, 560-594, 584-606, 611-645, 617-363, 623-642, 617-645, 6 39-754, 639-658, 639-654, 641-656, 642-657, 643-658, 642-754, 653-672, 662-685, 665685,665-689, 668-687, 670-754, 670-706, 670-685, 670-686, 670-689, 671-690, 671-691, 671-686, 671-687, 672-693, 672-697, 672-707, 672687, 672-688, 673-688, 674-693 , 678-693, 679-694, 679-707, 679-698, 679-701, 679-702, 679-707, 680-695, 680-699, 679-699, 681-706, 681696, 682-697, 682-7 06, 682-707, 682-702, 682-701, 683-698, 684-699, 685-700, 686-701, 687-754, 688-704, 689-709, 689-710, 690-705, 679705, 679 -710, 679-706, 690-710, 691-710, 690-754, 690-706, 684-703, 687-705, 687-702, 687-703, 687-706, 688-703, 688-704, 688-705, 689704, 689-705, 689-708, 690-705, 690-706, 690-709, 691-706, 692-711, 693-716, 693-712, 695-715, 697-716, 697-716, 690-716 . 73, 757-776, 757-785, 790815, 793-812, 811-833, 811-844, 814-833, 811-906, 820-839, 822-844, 822-867, 823-842, 845-864, 845 -867, 854-906, 845-909, 845-906, 854876, 863-882, 863-885, 878-900, 887-906, 899-918, 899-933, 899-958, 905-927, 905-933, 9 14-933, 936-958, 936-955, 945-964, 951-970, 951985, 951-1044, 951-1024, 951-1056, 951-997, 960-985, 963-1044, 963-1024, 963 -997, 972-1015, 1025-1044, 1031-1056, 1037-1056, 1046-1083, 1049-1068, 1070-1089, 1070-1095, 1082-1101, 10811134, 1081-1143 1082-1101 1088-1107 1088-1134 1094-1119 1097-1119 1112-1134 1118-1143 1118-1146 1088-1146 11211140 1127-1146 1 127-1193, 1150-1193, 1156-1187, 1165-1187, 1170-1192, 1171-1191, 1172-1191, 1176-1192, 1176-1285, 11771192, 1176-1191, 1203 -1297, 1206-1228, 1206-1255, 1209-1228, 1215-1255, 1245-1265, 1251-1280, 1262-1285, 1251-1285, 1259-1296, 1259-1290, 1259-12 87, 1261-1296, 1261-1285, 1261-1276, 1261-1277, 1261-1280, 1262-1296, 1262-1277, 1262-1278, 12621281, 1263-1278, 1263-1279, 1263-1282, 1264-1279, 1264-1280, 1264-1283, 1264-1297, 1265-1280, 1265-1281, 1265-1284, 12661281, 1266-1282, 1266-1285, 12 67-1282, 1267-1283, 1268-1283, 1268-1284, 1268-1296, 1269-1284, 1269-1285, 1269-1288, 12701285, 1271-1290, 1271-1296, 1277- 1296, 1261-1290, 1262-1290, 1268-1290, 1263-1305, 1259-1305, 1259-1305, 1266-1305, 12591302, 1275-1294, 1281-1306, 1281-132 4, 1281-1336, 1282-1301, 1286-1306, 1290-1324, 1293-1318, 1290-1324, 1293-1315, 12961315, 1311-1336, 1311-1333, 1326-1345, 1353-1381, 1359-1378, 1395-1414, 1498-1532, 1498-1523, 1498-1535, 1510-1529, 1515-1535, 1515-1563, 1515-1596, 1515-1605, 151 5-1602, 1515-1540, 1515-1535, 1518-1605, 1518-1602, 1518-1537, 1521-1563, 15211540, 1550-1655, 1550-1563, 1550-1569, 1553-1 578, 1553-1599, 1553-1590, 1565-1584, 1571-1595, 1577-1605, 1577-1606, 1577-1592, 1577-1593, 1577-1596, 1578-1593, 1578-1594 , 1578-1597, 1578-1598, 1571-1598, 1579-1594, 1579-1594, 1579-1598, 1580-1595, 1580-1596, 1580-1599, 1580-1605, 1580-1602, 1 581-1596, 1581-1597, 1581-1600, 1582-1597, 1582-1598, 1582-1601, 1582-1602, 1553-1655, 1583-1598, 1583-1599, 1583-1602, 1584 -1599, 1584-1600, 1584-1603, 1585-1600, 1585-1601, 1585-1604, 1586-1601, 1586-1602, 1586-1605, 1587-1602, 1587-1603, 1587-16 06, 1588-1603, 1588-1604, 1589-1604, 1589-1605, 1590-1605, 15901606, 1591-1606, 1586-1652, 1642-1664, 1651-1720, 1651-1673, 1655-1679, 1695-1720, 1716-1738, 1743-1763, 1743-1768, 1764-1783, 1773-1792, 1777-1796, 1777-1800, 1778-1800, 1778-1793, 17 78-1794, 1778-1797, 1779-1798, 1779-1797, 1779-1796, 1779-1795, 1779-1794, 1780-1799, 1780-1796, 1780-1795, 1781-1797, 1781- 1796, 1781-1796, 1781-1800, 1781-1797, 1782-1799, 1782-1797, 1782-1798, 1783-1798, 1783-1799, 1784-1800, 1784-1799, 1779-179 9, 1778-1889, 1778-1794, 1779-1795, 1780-1799, 1785-1800, 1794-1813, 1806-1837, 1806-1828, 1806-1825, 1809-1828, 1812-1843, 1812-1837, 1812-1831, 1815-1843, 1815-1844, 1815-1840, 1815-1834, 1818-1837, 1821-1840, 1821-1844, 1821-1837, 1822-1843, 182 2-1839, 1822-1837, 1823-1843, 1823-1838, 1824-1839, 1827-1846, 1861-1884, 1861-1880, 1865-1885, 1866-1881, 1867-1882, 1867-1 886, 1868-1883, 1869-1885, 1869-1884, 1870-1885, 1871-1886, 1872-1887, 1874-1889, 1876-1895, 1888-1914, 1888-1908, 1891-1910 , 1891-1914, 1895-1938, 1895-1935, 1913-1935, 1898-1920, 1907-1929, 1913-1935, 1918-1934, 1919-1938, 1919-1934, 1921-1934, 1 928-1956, 1957-1976, 2035-2057, 20832141, 2230-2261, 2368-2393, 2381-2397, 2368-2394, 2379-2394, 2381-2396, 2368-2397, 2368 -2396, 2420-2439, 2458-2476, 2459 2478, 2819-2838, 2818-2838, 2873-2892, and 3161-3182.

[000258] In certain embodiments, an antisense compound or oligonucleotide target to an HBV nucleic acid is complementary within the gene region of the second HBV pre-S1 portion corresponding to nucleotide region 1-1932 of SEQ ID NO: 1. In certain embodiments, an antisense compound or oligonucleotide target to an HBV nucleic acid is complementary within the gene region of the first HBV pre-S1 portion corresponding to nucleotide region 2 831-3182 of SEQ ID NO: 1.

[000259] In certain embodiments, an antisense compound or oligonucleotide target for an HBV nucleic acid targets the gene region of the second HBV pre-S1 portion corresponding to the nucleotide region 1-1932 of SEQ ID NO: 1. 8313182 of SEQ ID NO: 1.

[000260] In certain embodiments, antisense compounds or oligonucleotides target a region of an HBV nucleic acid. In certain embodiments, such compounds or oligonucleotides targeting a region of an HBV nucleic acid have a contiguous nucleobase portion that is complementary to a nucleobase portion of equal length as the region. For example, the moiety can be at least a moiety of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 contiguous nucleobases complementary to a moiety equal in length to a region referred to herein. In certain embodiments, such compounds or oligonucleotides target the following nucleotide regions of SEQ ID NO: 1: 1-20, 1029, 10-56, 13-38, 13-35, 19-38, 25-47, 25-50, 25-56, 43-68, 43-63, 5574, 58-73, 58- 74, 58-77, 58-79, 58-80, 58-84, 59-74, 59-75, 59-80, 6075, 60-76, 60-79, 61-76, 61-77, 61-80, 62-77, 63-84, 68-114, 101-123 2 18, 199-224, 200-224, 205-224, 206-228, 218-237, 224-243, 233-264, 242-263, 243262, 244-263, 245-274; 245-260, 245-264, 246-266, 247-266, 247-269, 247-270, 245-267, 251-267, 245-266, 250-269, 251-266, 251-268, 251269, 245-2 69 245-266 245-261 250-265 250-266 250-267 250-268 250-269 251-270 252-267 253-268 253-269 253-272 253-274 4269, 254-270, 254-274, 255-270, 255-271, 255-274, 255-401, 255-400, 255-274, 255-273, 255-272, 255-271, 256-271, 256-275, 255-276, 256272, 256-276, 253-275, 256-279, 257-276, 258-273, 259-274, 260-279, 262-281, 262-321, 262-315, 262-312, 265-312 . 21, 324-343, 339-361, 339-367, 348367, 342-367, 358-392, 358-378, 360-392, 360-383, 360-388, 360-385, 362-381, 366-388, 369 4 11-426, 411-427, 411-430, 411-431, 411-437, 412-431, 411426, 411-427, 412-428, 412-431, 412-427, 413-433, 413-432, 413-428, 413-429, 413-432, 413-433, 414-427, 415-427, 414-429, 414-430, 414433, 415-428, 415-429, 415-430, 415-431, 415-434, 416-43 1, 416-432, 416-429, 416-435, 417-432, 417-433, 417-436, 418-433, 418-435, 418434, 418-437, 419-435, 419-434, 420-435, 419- 432, 419-434, 421-436, 422-437, 422-441, 423-436, 425-465, 454-473, 454-472, 457-476, 457472, 457-473, 454-476, 455-472, 45 7-485, 458-485, 458-483, 458-477, 458-473, 459-485, 460-485, 463-498, 463-485, 466-485, 463-482, 457491, 458-491, 459-491, 460-491, 463-491, 466-491, 472-491, 472-493, 473-492, 475-491, 459-494, 460-494, 463-494, 466-494, 467-498, 472494, 475-494 ,457-473,457-472,458-494,454-494,457-494,457-473,485-513,470-493,476-519,485-519,500-519,512-534,512-550,5245 46, 536-559, 548-567, 548-570, 550-570, 548-594, 554-573, 548-576, 560-594, 584-606, 611-645, 617-363, 623-642, 617-645, 63 9-754, 639658, 639-654, 641-656, 642-657, 643-658, 642-754, 653-672, 662-685, 665-685,665-689, 668-687, 670-754, 670-706, 6 70-685, 670-686, 670689, 671-690, 671-691, 671-686, 671-687, 672-693, 672-697, 672-707, 672-687, 672-688, 673-688, 674-693, 678-693, 679-694, 679-707, 679698, 679-701, 679-702, 679-707, 680-695, 680-699, 679-699, 681-706, 681-696, 682-697, 682-70 6, 682-707, 682-702, 682-701, 683-698, 684699, 685-700, 686-701, 687-754, 688-704, 689-709, 689-710, 690-705, 679-705, 679- 710, 679-706, 690-710, 691-710, 690-754, 690-706, 684703, 687-705, 687-702, 687-703, 687-706, 688-703, 688-704, 688-705, 68 9-704, 689-705, 689-708, 690-705, 690-706, 690-709, 691-706, 692711, 693-716, 693-712, 695-715, 697-716, 697-716, 690-716, 724-746, 724-752, 724-754, 724-758, 733-752, 738-754, 738-753, 739-758, 739754, 739-775, 739-754, 740-754, 742-785, 742-773 , 757-776, 757-785, 790-815, 793-812, 811-833, 811-844, 814-833, 811-906, 820-839, 822844, 822-867, 823-842, 845-864, 845-8 67, 854-906, 845-909, 845-906, 854-876, 863-882, 863-885, 878-900, 887-906, 899-918, 899-933, 899958, 905-927, 905-933, 914 -933, 936-958, 936-955, 945-964, 951-970, 951-985, 951-1044, 951-1024, 951-1056, 951-997, 960-985, 963-1044, 963-1024, 963- 997, 972-1015, 1025-1044, 1031-1056, 1037-1056, 1046-1083, 1049-1068, 1070-1089, 1070-1095, 1082-1101, 10811134, 1081-1143, 1082-1101, 1088-1107, 1088-1134, 1094-1119, 1097-1119, 1112-1134, 1118-1143, 1118-1146, 1088-1146, 11211140, 1127-1146, 11 27-1193, 1150-1193, 1156-1187, 1165-1187, 1170-1192, 1171-1191, 1172-1191, 1176-1192, 1176-1285, 11771192, 1176-1191, 1203- 1297, 1206-1228, 1206-1255, 1209-1228, 1215-1255, 1245-1265, 1251-1280, 1262-1285, 1251-1285, 12591296, 1259-1290, 1259-128 7, 1261-1296, 1261-1285, 1261-1276, 1261-1277, 1261-1280, 1262-1296, 1262-1277, 1262-1278, 12621281, 1263-1278, 1263-1279, 1263-1282, 1264-1279, 1264-1280, 1264-1283, 1264-1297, 1265-1280, 1265-1281, 1265-1284, 12661281, 1266-1282, 1266-1285, 126 7-1282, 1267-1283, 1268-1283, 1268-1284, 1268-1296, 1269-1284, 1269-1285, 1269-1288, 12701285, 1271-1290, 1271-1296, 1277-1 296, 1261-1290, 1262-1290, 1268-1290, 1263-1305, 1259-1305, 1259-1305, 1266-1305, 12591302, 1275-1294, 1281-1306, 1281-1324 . 353-1381, 1359-1378, 1395-1414, 1498-1532, 1498-1523, 1498-1535, 1510-1529, 1515-1535, 1515-1563, 1515-1596, 1515-1605, 1515 -1602, 1515-1540, 1515-1535, 1518-1605, 1518-1602, 1518-1537, 1521-1563, 15211540, 1550-1655, 1550-1563, 1550-1569, 1553-15 78, 1553-1599, 1553-1590, 1565-1584, 1571-1595, 1577-1605, 1577-1606, 1577-1592, 1577-1593, 1577-1596, 1578-1593, 1578-1594, 1578-1597, 1578-1598, 1571-1598, 1579-1594, 1579-1594, 1579-1598, 1580-1595, 1580-1596, 1580-1599, 1580-1605, 1580-1602, 15 81-1596, 1581-1597, 1581-1600, 1582-1597, 1582-1598, 1582-1601, 1582-1602, 1553-1655, 1583-1598, 1583-1599, 1583-1602, 1584- 1599, 1584-1600, 1584-1603, 1585-1600, 1585-1601, 1585-1604, 1586-1601, 1586-1602, 1586-1605, 1587-1602, 1587-1603, 1587-16 06, 1588-1603, 1588-1604, 1589-1604, 1589-1605, 1590-1605, 1590-1606, 1591-1606, 1586-1652, 1642-1664, 1651-1720, 1651-1673 , 1655-1679, 1695-1720, 1716-1738, 1743-1763, 1743-1768, 1764-1783, 1773-1792, 1777-1796, 1777-1800, 1778-1800, 1778-1793, 1778-1794, 1778-1797, 1779-1798, 1779-1797, 1779-1796, 1779-1795, 1779-1794, 1780-1799, 1780-1796, 1780-1795, 1781-1797, 17 81-1796, 1781-1796, 1781-1800, 1781-1797, 1782-1799, 1782-1797, 1782-1798, 1783-1798, 1783-1799, 1784-1800, 1784-1799, 1779 -1799, 1778-1889, 1778-1794, 1779-1795, 1780-1799, 1785-1800, 1794-1813, 1806-1837, 1806-1828, 1806-1825, 1809-1828, 1812-1 843, 1812-1837, 1812-1831, 1815-1843, 1815-1844, 1815-1840, 1815-1834, 1818-1837, 1821-1840, 1821-1844, 1821-1837, 1822-184 3, 1822-1839, 1822-1837, 1823-1843, 1823-1838, 1824-1839, 1827-1846, 1861-1884, 1861-1880, 1865-1885, 1866-1881, 1867-1882, 1867-1886, 1868-1883, 1869-1885, 1869-1884, 1870-1885, 1871-1886, 1872-1887, 1874-1889, 1876-1895, 1888-1914, 1888-1908, 1 891-1910, 1891-1914, 1895-1938, 1895-1935, 1913-1935, 1898-1920, 1907-1929, 1913-1935, 1918-1934, 1919-1938, 1919-1934, 192 1-1934, 1928-1956, 1957-1976, 2035-2057, 2083-2141, 2230-2261, 2368-2393, 2381-2397, 2368-2394, 2379-2394, 2381-2396, 2368- 2397, 2368-2396, 2420-2439, 2458-2476, 2459 2478, 2819-2838, 2818-2838, 2873-2892, and 3161-3182.

[000261] In certain embodiments, antisense compounds or oligonucleotides target a region of an HBV nucleic acid. In certain embodiments, such compounds or oligonucleotides targeting a region of an HBV nucleic acid have a contiguous nucleobase portion that is complementary to a nucleobase portion of equal length as the region. For example, the moiety can be at least a moiety of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobases complementary to a moiety equal in length to a region referred to herein. In certain embodiments, such compounds or oligonucleotide target the following nucleotide regions of SEQ ID NO: 1: 58-73, 5874, 58-77, 59-74, 60-75, 61-76, 62-77, 245-274; 245-260, 250-265, 251-266, 252-267, 253-268, 254-269, 255-270, 256-271, 256-272, 258273, 259-274, 380-399, 382-401, 411-4 37, 411-427, 411-426, 412-427, 413-428, 413-432, 414-429, 415-430, 416-431, 417-432, 418-433, 419434, 420-435, 421-436, 422 -437, 457-472, 458-473, 639-754, 639-658, 639-654, 641-656, 642-657, 643-658, 670-754, 670-706, 670-685, 671686, 672-687, 6 73-688, 678-693, 679-694, 680-695, 681-706, 681-696, 682-697, 683-698, 684-699, 685-700, 686-701, 687-702, 688-703, 689704, 1 264-1279, 1265-1280, 1266-1281, 1267-1282, 1268-1283, 1269-1284, 1270-1285, 1577-1606, 1577-1592, 1578-1593, 1579-1594, 1580 -1595, 1581-1596, 1582-1597, 1583-1598, 1584-1599, 1585-1600, 1586-1601, 1587-1602, 1588-1603, 1589-1604, 1590-1605, 1591-16 06, 1778-1889, 1778-1800, 1778-1793, 1779-1794, 1780-1799, 1780-1796, 1780-1795, 1781-1796, 1782-1797, 1783-1798, 1784-1799, 1785-1800, 1822-1839, 1822-1837, 1823-1838, 1824-1839, 1866-1881, 1867-1882, 1868-1883, 1869-1884, 1870-1885, 1871-1886, 18 72-1887, or 1874-1889, and wherein at least one nucleoside of the compound or modified oligonucleotide comprises at least one 2'-O-methoxyethyl or restricted ethyl (cEt) sugar.

[000262] In certain embodiments, an antisense compound or target oligonucleotide for an HBV nucleic acid is complementary within the following nucleotide regions of SEQ ID NO: 1: 58-73, 58-74, 58-77, 59-74, 59-75, 60-75, 60-76, 61-76, 61-77, 62-77, 253-272, 253-269, 254-270, 255-271, 256-272, 411-437, 411-426, 411-427, 411-430, 412-427, 412-428, 412-431, 413-428, 413-4 29, 413-432, 414429, 414-430, 414-433, 415-430, 415-431, 415-434, 416-431, 416-432, 416-435, 417-432, 417-433, 417-436, 418 -433, 418-434, 418-437, 457472, 457-473, 458-473, 670-706, 670-685, 670-686, 671-686, 671-687, 672-687, 672-688, 673-688, 6 87-702, 687-703, 687-706, 688-703, 688704, 689-704, 689-705, 690-705, 690-706, 691-706, 1261-1285, 12611276, 1261-1277, 126 1-1280, 1262-1277, 1262-1278, 1262-1281, 1263-1278, 1263-1279, 1263-1282, 1264-1279, 1264-1280, 12641283, 1265-1280, 1265-1 281, 1265-1284, 1266-1281, 1266-1282, 1266-1285, 1267-1282, 1267-1283, 1268-1283, 1268-1284, 1269-1284, 1269-1285, 1270-1285 . 580-1596, 1580-1599, 1581-1596, 1581-1597, 1581-1600, 1582-1597, 1582-1598, 1582-1601, 1583-1598, 1583-1599, 1583-1602, 1584 -1599, 1584-1600, 1584-1603, 1585-1600, 1585-1601, 1585-1604, 1586-1601, 1586-1602, 1586-1605, 1587-1602, 1587-1603, 1587-16 06, 1588-1603, 1588-1604, 1589-1604, 1589-1605, 1590-1605, 1590-1606, 1591-1606, 1778-1800, 1778-1793, 1778-1794, 1778-1797, 1779-1794, 1779-1795, 1779-1798, 1780-1795, 1780-1796, 1780-1799, 1781-1796, 17811797, 1781-1800, 1782-1797, 1782-1798, 17 83-1798, 1783-1799, 1784-1799, and 1784-1800.

[000263] In certain embodiments, an antisense compound or oligonucleotide targeting an HBV nucleic acid targets the following nucleotide regions of SEQ ID NO: 1: 58-73, 58-74, 5877, 59-74, 59-75, 60-75, 60-76, 61-76, 61-77, 62-77, 253 -272, 253269, 254-270, 255-271, 256-272, 411-437, 411-426, 411-427, 411-430, 412-427, 412-428, 412-431, 413-428, 413-429, 4 13-432, 414-429, 414430, 414-433, 415-430, 415-431, 415-434, 416-431, 416-432, 416-435, 417-432, 417-433, 417-436, 418-433, 418-434, 418-437, 457-472, 457473, 458-473, 670-706, 670-685, 670-686, 671-686, 671-687, 672-687, 672-688, 673-688, 687-70 2, 687-703, 687-706, 688-703, 688-704, 689704, 689-705, 690-705, 690-706, 691-706, 1261-1285, 1261-1276, 1261-1277, 1261-12 80, 1262-1277, 1262-1278, 1262-1281, 1263-1278, 1263-1279, 1263-1282, 1264-1279, 1264-1280, 1264-1283, 1265-1280, 1265-1281, 1265-1284, 1266-1281, 1266-1282, 1266-1285, 1267-1282, 1267-1283, 1268-1283, 1268-1284, 1269-1284, 1269-1285, 1270-1285, 15 77-1606, 1577-1592, 1577-1593, 1577-1596, 1578-1593, 1578-1594, 1578-1597, 1579-1594, 1579-1594, 1579-1598, 1580-1595, 1580- 1596, 1580-1599, 1581-1596, 1581-1597, 1581-1600, 1582-1597, 1582-1598, 1582-1601, 1583-1598, 1583-1599, 1583-1602, 1584-159 9, 1584-1600, 1584-1603, 1585-1600, 1585-1601, 1585-1604, 1586-1601, 1586-1602, 1586-1605, 1587-1602, 1587-1603, 1587-1606, 1588-1603, 1588-1604, 1589-1604, 1589-1605, 1590-1605, 1590-1606, 1591-1606, 1778-1800, 1778-1793, 1778-1794, 1778-1797, 177 9-1794, 1779-1795, 1779-1798, 1780-1795, 1780-1796, 1780-1799, 1781-1796, 1781-1797, 1781-1800, 1782-1797, 1782-1798, 1783-1 798, 1783-1799, 1784 1799, and 1784-1800.

[000264] In certain embodiments, antisense compounds or oligonucleotides target a region of an HBV nucleic acid. In certain embodiments, such compounds or oligonucleotides targeting a region of an HBV nucleic acid have a contiguous nucleobase portion that is complementary to a nucleobase portion of equal length as the region. For example, the moiety can be at least a moiety of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobases complementary to a moiety equal in length to a region referred to herein. In certain embodiments, such compounds or oligonucleotides target the following nucleotide regions of SEQ ID NO: 1: 1-20, 1029, 10-56, 13-38, 13-35, 19-38, 25-47, 25-50, 25-56, 43-68, 43-63, 5574, 58-73, 58- 74, 58-77, 58-79, 58-80, 58-84, 59-74, 59-75, 59-80, 6075, 60-76, 60-79, 61-76, 61-77, 61-80, 62-77, 63-84, 68-114, 101-123 2 18, 199-224, 200-224, 205-224, 206-228, 218-237, 224-243, 233-264, 242-263, 243262, 244-263, 245-264, 246-266, 247-266, 247 -269, 247-270, 245-267, 251-267, 245-266, 250-269, 251-268, 251-269, 245-269, 245-266, 245261, 250-266, 250-267, 250-268, 2 50-269, 251-270, 253-269, 253-272, 253-274, 254-270, 254-274, 255-271, 255-274, 255-401, 255-400, 255274, 255-273, 255-272, 255-271, 256-275, 255-276, 256-272, 256-276, 253-275, 256-279, 257-276, 260-279, 262-281, 262-321, 262-315, 262312, 265-31 2, 266-288, 266-291, 266-285, 281-321, 281-303, 290-321, 290-312, 292-311, 290-312, 293-312, 293-315, 293-321, 296-321, 302 321, 324-343, 339-361, 339-367, 348-367, 342-367, 358-392, 358-378, 360-392, 360-383, 360-388, 360-385, 362-381, 366-388, 3 69-388, 366385, 366-392, 370-389, 370-392, 384-433, 384-400, 384-401, 385-401, 405-424, 409-428, 405-428, 411-426, 411-427, 411-430, 411-431, 411437, 412-431, 411-426, 411-427, 412-428, 412-431, 412-427, 413-433, 413-432, 413-428, 413-429, 413-43 2, 413-433, 414-427, 415-427, 414429, 414-430, 414-433, 415-428, 415-429, 415-430, 415-431, 415-434, 416-431, 416-432, 416- 429, 416-435, 417-432, 417-433, 417-436, 418433, 418-435, 418-434, 418-437, 419-435, 419-434, 420-435, 419-432, 419-434, 42 2-441, 423-436, 425-465, 454-473, 454-472, 457-476, 457472, 457-473, 454-476, 455-472, 457-485, 458-485, 458-483, 458-477, 458-473, 459-485, 460-485, 463-498, 463-485, 466-485, 463-482, 457491, 458-491, 459-491, 460-491, 463-491, 466-491, 472-491 ,472-493,473-492,475-491,459-494,460-494,463-494,466-494,467-498,472494,475-494,457-473,457-472,458-494,454-4 94, 457-494, 457-473, 485-513, 470-493, 476-519, 485-519, 500-519, 512-534, 512-550, 524546, 536-559, 548-567, 548-570, 550 -570, 548-594, 554-573, 548-576, 560-594, 584-606, 611-645, 617-363, 623-642, 617-645, 642-754, 653672, 662-685, 665-685, 6 65-689, 668-687, 670-706, 670-685, 670-686, 670-689, 671-690, 671-691, 671-686, 671-687, 672-693, 672-697, 672707, 672-687, 672-688, 673-688, 674-693, 679-707, 679-698, 679-701, 679-702, 679-707, 680-699, 679-699, 682-706, 682-707, 682-702, 68270 1, 687-754, 688-704, 689-709, 689-710, 690-705, 679-705, 679-710, 679-706, 690-710, 691-710, 690-754, 690-706, 684-703, 687 -705, 687702, 687-703, 687-706, 688-703, 688-704, 688-705, 689-704, 689-705, 689-708, 690-705, 690-706, 690-709, 691-706, 6 92-711, 693-716, 693712, 695-715, 697-716, 697-716, 690-716, 724-746, 724-752, 724-754, 724-758, 733-752, 738-754, 739-758, 739-754, 739-775, 739-754, 740754, 742-785, 742-773, 757-776, 757-785, 790-815, 793-812, 811-833, 811-844, 814-833, 811-90 6, 820-839, 822-844, 822-867, 823-842, 845864, 845-867, 854-906, 845-909, 845-906, 854-876, 863-882, 863-885, 878-900, 887- 906, 899-918, 899-933, 899-958, 905-927, 905-933, 914 933, 936-958, 936-955, 945-964, 951-970, 951-985, 951-1044, 9511024, 1 070-1089, 1070-1095, 1082-1101, 1081-1134, 1081-1143, 10821101, 1088-1107, 1088-1134, 1094-1119, 1097-1119, 1112-1134, 1118 -1143, 1118-1146, 1088-1146, 1121-1140, 1127-1146, 1127-1193, 1150-1193, 1156-1187, 1165-1187, 1170-1192, 1171-1191, 1172-11 91, 1176-1192, 1177-1192, 1176-1191, 1203-1297, 1206-1228, 1206-1255, 1209-1228, 1215-1255, 1245-1265, 1251-1280, 1262-1285, 1251-1285, 1259-1296, 1259-1290, 1259-1287, 12611296, 1261-1285, 1261-1276, 1261-1277, 1261-1280, 1262-1296, 1262-1277, 12 62-1278, 1262-1281, 1263-1278, 1263-1279, 1263-1282, 1264-1279, 1264-1280, 1264-1283, 1264-1297, 1265-1280, 1265-1281, 1265- 1284, 1266-1281, 1266-1282, 1266-1285, 1267-1282, 1267-1283, 1268-1283, 1268-1284, 1268-1296, 1269-1284, 1269-1285, 1269-128 8, 1270-1285, 1271-1290, 1271-1296, 12771296, 1261-1290, 1262-1290, 1268-1290, 1263-1305, 1259-1305, 1259-1305, 1266-1305, 1259-1302, 1275-1294, 1281-1306, 12811324, 1281-1336, 1282-1301, 1286-1306, 1290-1324, 1293-1318, 1290-1324, 1293-1315, 129 6-1315, 1311-1336, 1311-1333, 13261345, 1353-1381, 1359-1378, 1395-1414, 1498-1532, 1498-1523, 1498-1535, 1510-1529, 1515-1 535, 1515-1563, 1515-1596, 1515-1605, 1515-1602, 1515-1540, 1515-1535, 1518-1605, 1518-1602, 1518-1537, 1521-1563, 1521-1540 1550-1655, 1550-1563, 1550-1569, 1553-1578, 1553-1599, 1553-1590, 1565-1584, 1571-1595, 1577-1605, 1577-1606, 1577-1592 577-1593, 1577-1596, 1578-1593, 1578-1594, 1578-1597, 1578-1598, 1571-1598, 1579-1594, 1579-1594, 1579-1598, 1580-1595, 1580 -1596, 1580-1599, 1580-1605, 1580-1602, 1581-1596, 1581-1597, 1581-1600, 1582-1597, 1582-1598, 1582-1601, 1582-1602, 1553-16 55, 1583-1598, 1583-1599, 1583-1602, 1584-1599, 1584-1600, 1584-1603, 1585-1600, 1585-1601, 1585-1604, 1586-1601, 1586-1602, 1586-1605, 1587-1602, 1587-1603, 1587-1606, 1588-1603, 1588-1604, 1589-1604, 1589-1605, 1590-1605, 1590-1606, 1591-1606, 15 86-1652, 1642-1664, 1651-1720, 1651-1673, 1655-1679, 1695-1720, 1716-1738, 1743-1763, 1743-1768, 1764-1783, 1773-1792, 1777- 1796, 1777-1800, 1778-1800, 1778-1793, 1778-1794, 1778-1797, 1779-1798, 1779-1797, 1779-1796, 1779-1795, 1779-1794, 1780-179 9, 1780-1796, 1780-1795, 1781-1797, 1781-1796, 1781-1796, 1781-1800, 1781-1797, 1782-1799, 1782-1797, 1782-1798, 1783-1798, 1783-1799, 1784-1800, 1784-1799, 1779-1799, 1778-1794, 1779-1795, 1780-1799, 1794-1813, 1806-1837, 1806-1828, 1806-1825, 180 91828, 1812-1843, 1812-1837, 1812-1831, 1815-1843, 1815-1844, 1815-1840, 1815-1834, 1818-1837, 1821-1840, 1821-1844, 182118 37, 1822-1843, 1822-1839, 1823-1843, 1827-1846, 1861-1884, 1861-1880, 1865-1885, 1867-1886, 1869-1885, 1876-1895, 18881914, 1888-1908, 1891-1910, 1891-1914, 1895-1938, 1895-1935, 1913-1935, 1898-1920, 1907-1929, 1913-1935, 1918-1934, 19191938, 19 19-1934, 1921-1934, 1928-1956, 1957-1976, 2035-2057, 2083-2141, 2230-2261, 2368-2393, 2381-2397, 2368-2394, 23792394, 2381- 2396, 2368-2397, 2368-2396, 2420-2439, 2458-2476, 2459-2478, 2819-2838, 2818-2838, 2873-2892, and 3161-3182.

[000265] In certain embodiments, antisense compounds or oligonucleotides target a region of an HBV nucleic acid. In certain embodiments, such compounds or oligonucleotides targeting a region of an HBV nucleic acid have a contiguous nucleobase portion that is complementary to a nucleobase portion of equal length as the region. For example, the moiety can be at least a moiety of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobases complementary to a moiety equal in length to a region referred to herein. In certain embodiments, such compounds or oligonucleotides target the following nucleotide regions of SEQ ID NO: 1: 233-264, 242-263, 243-262, 244-263, 245-264, 246-266, 247-266, 247-269, 247270, 245-267, 25 1-267, 245-266, 250-269, 251-268, 251-269, 245-269, 245-266, 245-261, 250-266, 250-267, 250-268, 250-269, 251-270, 253272, 253-274, 254-274, 255-274, 255-401, 255-400, 255-274, 255-273, 255-272, 255-271, 256-275, 255-276, 256-276, 253-275, 256-27 9, 257276, 260-279, 262-281, 262-321, 262-315, 262-312, 265-312, 266-288, 266-291, 266-285, 281-321, 281-303, 405-424, 409- 428, 405-428, 411-430, 411-431, 411-431, 412-431, 411-426, 411-427, 412-428, 412-431, 412-427, 413-433, 413-432, 413-428, 41 3-433, 411-427, 414-427, 415427, 415-428, 415-429, 416-432, 416-429, 418-435, 418-434, 419-435, 419-434, 420-435, 419-432, 419-434, 422-441, 423-436, 425-465, 584606, 611-645, 617-363, 623-642, 617-645, 642-754, 653-672, and wherein at least one nucleoside of the compound or modified oligonucleotide comprises at least one 2' sugar -O-methoxyethyl.

[000266] In certain embodiments, the following nucleotide regions of SEQ ID NO: 1, when targeted by antisense compounds or oligonucleotides, exhibit at least 50% inhibition: 1-20, 10-29, 10-56, 13-38, 13-35, 19-38, 25-47, 25-50, 25-56, 43- 68, 43-63, 55-74, 58-77, 58-74, 58-73, 58-79, 58-80, 58-84, 59-74, 59-75, 59-80, 60-79, 60-75, 60-76, 61-80, 61-76, 61-77, 62-77, 63-84, 68-114, 101-123, 98-123, 113-138, 116-138, 131-150, 137-162, 152-186, 158177, 167-186, 191-215, 196-224, 196- 215, 196-218, 199-228, 199-218, 199-224, 200-224, 205-224, 206-228, 218-237, 224-243, 233-264, 242263, 243-262, 244-263, 24 5-264, 246-266, 247-266, 247-269, 247-270, 245-267, 251-267, 245-266, 250-269, 251-268, 251-269, 245-269, 245266, 245-261, 250-265, 250-266, 250-267, 250-268, 250-269, 251-266, 251-270, 252-267, 253-268, 253-269, 253-272, 253-274, 254-269, 254270 ,254-274,255-274,255-401,255-400,255-274,255-273,255-272,255-271,255-270,256-271,256-272,256-275,255-276,256- 276, 253275, 256-279, 257-276, 258-273, 259-274, 260-279, 262-281, 262-321, 262-315, 262-312, 265-312, 266-288, 266-291, 26 6-285, 281-321, 281303, 290-321, 290-312, 292-311, 290-312, 293-312, 293-315, 293-321, 296-321, 302-321, 324-343, 339-361, 339-367, 348-367, 342-367, 358392, 358-378, 360-392, 360-383, 360-388, 360-385, 362-381, 366-388, 369-388, 366-385, 366-392 , 370-389, 370-392, 380-399, 382-401, 384433, 384-400, 384-401, 385-401, 405-424, 409-428, 405-428, 411-430, 411-431, 411-4 31, 412-431, 411-426, 411-427, 411-430, 411-437, 412428, 412-431, 412-427, 413-432, 413-428, 413-429, 413-433, 411-427, 414 -427, 414-429, 414-430, 414-433, 415-427, 415-428, 415-429, 415430, 415-431, 415-434, 416-435, 416-432, 416-431, 416-429, 4 17-432, 417-433, 417-436, 418-437, 418-435, 418-434, 418-433, 419-435, 419434, 420-435, 419-432, 419-434, 421-436, 422-441, 422-437, 423-436, 425-465, 454-473, 454-472, 457-476, 454-476, 455-472, 457-485, 457473, 457-472, 458-485, 458-483, 458-47 7, 458-473, 459-485, 460-485, 463-498, 463-485, 466-485, 463-482, 457-491, 458-491, 459-491, 460491, 463-491, 466-491, 472- 491, 472-493, 473-492, 475-491, 459-494, 460-494, 463-494, 466-494, 467-498, 472-494, 475-494, 457-473, 457472, 458-494, 45 4-494, 457-494, 457-473, 485-513, 470-493, 476-519, 485-519, 500-519, 512-534, 512-550, 524-546, 536-559, 548-567, 548570, 550-570, 548-594, 554-573, 548-576, 560-594, 584-606, 611-645, 617-363, 623-642, 617-645, 639-654, 641-656, 642-657, 642-75 4, 643 658, 653-672, 662-685, 665-685,665-689, 668-687, 670-689, 670-706, 670-685, 670-686, 671-686, 671-687, 671-690, 671- 691, 672-687, 672688, 672-693, 672-697, 672-707, 673-688, 674-693, 678-693, 679-707, 679-694, 679-698, 679-701, 679-702, 67 9-707, 680-699, 679-699, 680695, 681-696, 682-706, 682-707, 682-702, 682-701, 682-697, 683-698, 684-699, 685-700, 686-701, 687-702, 687-703, 687-706, 687-754, 688703, 688-704, 689-704, 689-705, 689-709, 689-710, 690-705, 690-706, 691-706, 679-705 . 11 693-716 693-712 695-715 697-716 697-716 690-716 724746 724-752 724-754 724-758 733-752 738-753 738-754 739 -758, 739-754, 739-775, 739-754, 740-754, 742-785, 742-773, 757-776, 757785, 790-815, 793-812, 811-833, 811-844, 814-833, 8 11-906, 820-839, 822-844, 822-867, 823-842, 845-864, 845-867, 854-906, 845-909, 845906, 854-876, 854-873, 863-882, 863-885, 878-900, 878-897, 887-906, 899-918, 899-933, 899-958, 905-927, 905-933, 914-933, 936-958, 936 955, 945-964, 951-970, 951-9 85, 951-1044, 951-1024, 951-1056, 951997, 960-985, 963-1044, 963-1024, 963-997, 966-985, 972-1015, 978997, 1025-1044, 1031- 1056, 1037-1056, 1046-1083, 1049-1068, 1070-1089, 1070-1095, 1082-1101, 1081-1134, 1081-1143, 1082-1101, 1088-1107, 1088-1 134, 1094-1119, 1097-1119, 1112-1134, 1118-1143, 1118-1146, 1088-1146, 1121-1140, 1127-1146, 1127-1193, 1150-1193, 1156-1187 1165-1187 1170-1192 1171-1191 11721191 1176-1192 1177-1192 1176-1191 1203-1297 1206-1228 1206-1255 1209-1228 1 215-1255, 1215-1234, 1218-1237, 12211240, 1224-1243, 1227-1246, 1230-1249, 1233-1252, 1236-1255, 1245-1265, 1251-1280, 1251 -1285, 1251-1270, 1254-1273, 1254-1279, 1257-1276, 1258-1277, 1259-1296, 1259-1290, 1259-1287, 1260-1279, 1261-1285, 1261-12 76, 1261-1277, 1261-1280, 12611296, 1262-1277, 1262-1278, 1262-1281, 1262-1285, 1262-1296, 1263-1278, 1263-1279, 1263-1282, 1264-1279, 1264-1280, 1264-1283, 1264-1297, 1265-1280, 1265-1281, 1265-1284, 1266-1281, 1266-1282, 1266-1285, 1267-1282, 12 67-1283, 1268-1283, 1268-1284, 1268-1296, 1269-1284, 1269-1285, 1269-1288, 1270-1285, 1271-1290, 1271-1296, 1277-1296, 1261- 1290, 1262-1290, 1268-1290, 1263-1305, 1259-1305, 1259-1305, 1266-1305, 1259-1302, 1275-1294, 1281-1306, 1281-1324, 1281-133 6, 1782-1797, 1282-1301, 1286-1306, 1290-1324, 1293-1318, 1290-1324, 1293-1315, 1296-1315, 1311-1336, 1311-1333, 1326-1345, 1353-1381, 13591378, 1395-1414, 1498-1532, 1498-1523, 1498-1535, 1510-1529, 1515-1535, 1515-1563, 1515-1596, 1515-1605, 151 5-1602, 1515-1540, 1515-1535, 1518-1605, 1518-1602, 1518-1537, 1521-1563, 1521-1540, 1550-1655, 1550-1563, 1550-1569, 1553-1 578, 1553-1599, 1553-1590, 1565-1584, 1571-1595, 1577-1606, 1577-1605, 1577-1596, 1577-1592, 1577-1593, 1578-1593, 1578-1594 , 1578-1597, 1578-1598, 1579-1594, 1579-1595, 1579-1598, 1571-1598, 1580-1605, 1580-1602, 1580-1595, 1580-1596, 1580-1599, 1 5811596, 1581-1597, 1581-1600, 1582-1597, 1582-1598, 1582-1601, 1582-1602, 1553-1655, 1583-1598, 1583-1599, 1583-1602, 1584 - 1599, 1584-1600, 1584-1603, 1585-1600, 1585-1601, 1585-1604, 1586-1601, 1586-1602, 1586-1605, 1586-1652, 1587-1602, 15871 603, 1587-1606, 1588-1603, 1588-1604, 1589-1604, 1589-1605, 1590-1605, 1590-1606, 1591-1606, 1642-1664, 1651-1720, 16511673 1655-1679, 1695-1720, 1716-1738, 1743-1763, 1743-1768, 1764-1783, 1773-1792, 1777-1796, 1777-1800, 1778-1800, 17781793 778-1794, 1778-1797, 1779-1798, 1779-1797, 1779-1796, 1779-1795, 1779-1794, 1780-1796, 1781-1797, 1781-1796, 1781-1800, 1781 -1797, 1782-1799, 1784-1800, 1779-1799, 1778-1794, 1779-1795, 1780-1799, 1794-1813, 1780-1795, 1780-1796, 1780-1799, 1781-17 96, 1781-1797, 1781-1800, 1782-1798, 1783-1799, 1784-1799, 1784-1800, 1785-1800, 1806-1837, 1806-1828, 1806-1825, 1809-1828, 1812-1843, 1812-1837, 1812-1831, 1815-1843, 1815-1844, 1815-1840, 1815-1834, 1818-1837, 1821-1840, 1821-1844, 1821-1837, 18 22-1843, 1822-1839, 1823-1843, 1827-1846, 1861-1884, 1861-1880, 1865-1885, 1867-1886, 1869-1885, 1876-1895, 1888-1914, 1888- 1908, 1891-1910, 1891-1914, 1895-1938, 1895-1935, 1913-1935, 1898-1920, 1907-1929, 1913-1935, 1918-1934, 1919-1938, 1919-193 4, 1921-1934, 1928-1956, 1957-1976, 2035-2057, 2083-2141, 2230-2261, 2278-2297, 2281-2300, 22842303, 2368-2393, 2381-2397, 2368-2394, 2379-2394, 2381-2396, 2368-2397, 2368-2396, 2420-2439, 2458-2476, 2459-2478, and 3 161-3182.

[000267] In certain embodiments, the following nucleotide regions of SEQ ID NO: 1, when targeted by antisense compounds or oligonucleotides, exhibit at least 60% inhibition: 1-20, 10-29, 10-53, 13-38, 25-50, 43-68, 55-74, 58-84, 58-77, 58- 74, 58-73, 58-79, 59-80, 59-74, 59-75, 60-75, 60-76, 61-77, 61-76, 61-80, 62-77, 68-114, 98-123, 101-123, 113-138, 116-138, 131-150, 137-162, 152-186, 191-215, 196-224, 196-215, 199-228, 199-218, 200-223, 199-218, 205-224, 206-228, 218-237, 224- 243, 233-263, 244-263, 245-264, 247-266, 250-265, 251-266, 252-267, 253-272, 253-269, 251-267, 253274, 254-270, 255-276, 25 6-279, 256-276, 256-274, 256-272, 256-271, 258-273, 259-274, 265-388, 265-284, 266-291, 266-288, 260-279, 281321, 281-303, 290-321, 290-312, 293-312, 296-315, 302-321, 324-343, 339-367, 339-361, 342-367, 348-367, 358-392, 358-378, 360-392, 360379 ,366-392,366-385,369-388,370-392,382-401,405-428,405-424,409-428,411-436,411-433,411-431,411-426,411-430,411- 427, 412431, 412-428, 412-427, 413-428, 413-429, 413-433, 414-433, 414-430, 414-429, 414-433, 415-430, 415-431, 415-434, 41 5-435, 415-436, 416-429, 416-434, 416-431, 416-432, 416-436, 416-435, 417-436, 417-433, 417-432, 418-434, 418-433, 418-437, 419-434, 420-435, 421-436, 422437, 423-436, 425-465, 454-472, 455-472, 457-476, 457-472, 457-473, 458-485, 458-473, 458-483 . 76, 560-594, 584-606, 608-648, 639-654, 640-656, 641656, 642-657, 642-658, 643-658, 653-672, 662-685, 665-685, 670-706, 670 -689, 670-685, 670-686, 671-690, 671-686, 671-687, 672-707, 672697, 672-693, 672-687, 672-688, 673-688, 679-707, 679-698, 6 79-694, 680-695, 681-696, 682-697, 682-701, 683-698, 684-699, 685-700, 686701, 687-754, 687-702, 687-705, 687-703, 687-706, 688-704, 688-703, 688-704, 688-705, 688-707, 689-710, 689-709, 689-705, 689-704, 690754, 690-705, 690-706, 691-706, 691-71 0, 692-711, 697-716, 724-758, 724-754, 724-752, 724-746, 738-754, 738-753, 739-754, 742-785, 757785, 790-815, 811-906, 811- 844, 811-833, 822-867, 822-844, 823-842, 845-867, 854-906, 854-873, 878-897, 899-958, 899-933, 936-958, 945964, 951-1044, 9 51-1024, 951-985, 951-997, 963-1044, 963-1024, 963-997, 966-985, 978-997, 1031-1056, 1046-1083, 1070-1095, 10811143, 1081-1 134, 1082-1101, 1088-1146, 1088-1134, 1118-1146, 1118-1143, 1127-1193, 1170-1189, 1176-1192, 1176-1191, 1177-1192, 1203-12 97, 1206-1255, 1209-1228, 1215-1234, 1218-1237, 1221-1240, 1224-1243, 1227-1246, 1230-1249, 1233-1252, 12361255, 1251-1270, 1251-1285, 1254-1273, 1254-1279, 1257-1276, 1258-1277, 1259-1278, 1260-1279, 1261-1276, 1261-1285, 12611276, 1261-1277, 12 61-1280, 1262-1281, 1262-1277, 1262-1278, 1262-1281, 1263-1278, 1263-1279, 1263-1282, 1264-1297, 1264-1279, 1264-1280, 1264- 1283, 1265-1280, 1265-1281, 1265-1284, 1266-1281, 1266-1282, 1266-1285, 1267-1282, 1267-1283, 12681283, 1268-1284, 1268-128 7, 1269-1284, 1269-1285, 1270-1285, 1281-1336, 1281-1324, 1281-1306, 1286-1305, 1290-1324, 13111336, 1326-1345, 1353-1381, 1395-1414, 1498-1535, 1498-1532, 1515-1535, 1515-1534, 1521-1540, 1550-1655, 1553-1599, 1553-1590, 1577-1606, 1577-1592, 157 7-1593, 1577-1596, 1578-1593, 1578-1594, 1578-1597, 1579-1594, 1579-1595, 1579-1598, 1580-1595, 1580-1596, 1580-1599, 1581-1 596, 1581-1597, 1581-1600, 1582-1597, 1582-1598, 1582-1601, 1583-1598, 1583-1599, 1583-1602, 1584-1599, 1584-1600, 1584-1603 . 589-1604, 1589-1605, 1590-1605, 1590-1606, 1591-1606, 1642-1664, 1651-1720, 1716-1738, 1743-1763, 1764-1783, 1773-1792, 1777 -1800, 1777-1797, 1655-1674, 1778-1794, 1778-1800, 1781-1800, 1781-1797, 1784-1800, 1779-1799, 1778-1794, 1778-1797, 1779-17 95, 1779-1798, 1780-1795, 1780-1796, 1780-1799, 1781-1796, 1781-1797, 17811800, 1782-1797, 1794-1813, 1806-1837, 1806-1825, 1812-1837, 1812-1831, 1815-1844, 1815-1834, 1818-1837, 1821-1837, 1822-1838, 1827-1846, 1861-1884, 1821-1840, 1866-1885, 18 67-1886, 1888-1914, 1888-1907, 1891-1914, 1895-1938, 1895-1935, 1919-1938, 1928-1956, 1957-1976, 2035-2057, 2083-2141, 2230- 2261, 2278-2297, 2281-2300, 2284-2303, 2368-2397, 2368-2396, 2368-2394, 2368-2393, 2379-2394, 2381-2396, 2420-2439, 2458-2 476, 2819-2838, 2873-2892, and 3161-3182.

[000268] In certain embodiments, the following nucleotide regions of SEQ ID NO: 1, when targeted by antisense compounds or oligonucleotides, exhibit at least 65% inhibition: 1-20, 10-29, 10-53, 13-38, 25-50, 43-68, 55-74, 58-84, 58-79, 58- 74, 58-73, 58-77, 59-75, 59-80, 58-77, 60-75, 60-76, 61-77, 61-76, 61-80, 62-77, 68-114, 98-123, 101-123, 113-138, 116-138, 131-150, 137-162, 152-186, 191-215, 196-215, 199-228, 199-218, 200-223, 199-218, 205224, 206-228, 218-237, 224-243, 233-263 ,244-263,245-264,250-265,251-266,253-269,253-274,255-276,256-279,256-276,256-274,256272,256-271,247-266,253-2 72, 258-273, 266-291, 266-288, 260-279, 281-321, 281-303, 290-321, 290-312, 296-315, 293-312, 302-321, 324343, 339-367, 339 -361, 342-367, 348-367, 358-392, 358-378, 360-392, 360-379, 366-392, 366-385, 369-388, 370-392, 382-401, 405-428, 405424, 4 09-428, 411-433, 411-431, 411-430, 411-427, 411-426, 412-431, 412-428, 412-427, 413-433, 413-428, 413-429, 413-432, 414-433 . 33, 417-432, 418-433, 418-434, 418-437, 420-435, 422-437, 423-436, 425-465, 454-472, 455-472, 457-472, 458-485, 458-483, 458 -473, 463-498, 467-498, 457476, 470-493, 472-491, 485-519, 485-513, 500-519, 512-534, 524-546, 536-558, 548-567, 554-573, 5 48-576, 560-594, 584-606, 608-648, 639654, 640-656, 641-656, 642-657, 642-658, 643-658, 653-672, 662-685, 665-685, 670-685, 670-706, 670-689, 670-686, 670-685, 671-686, 671687, 671-690, 672-688, 672-687, 672-707, 672-697, 672-693, 673-688, 679-69 8, 680-695, 681-696, 682-697, 682-701, 683-698, 684-699, 685700, 686-701, 687-702, 688-703, 688-707, 687-754, 690-754, 690- 706, 690-705, 687-705, 687-703, 687-706, 687-702, 688-705, 688-703, 688704, 689-705, 691-706, 692-711, 697-716, 724-758, 72 4-754, 724-752, 724-746, 738-754, 739-754, 742-785, 757-785, 790-815, 811-906, 811-844, 811-833, 822-867, 822-844, 823-842, 845-867, 854-906, 854-873, 878-897, 899-958, 899-933, 936-958, 945-964, 951-1044, 951-1024, 951-985, 951-997, 963-1044, 963 -1024, 963-997, 966-985, 978-997, 1031-1056, 1046-1083, 1070-1095, 1081-1143, 1081-1134, 10821101, 1088-1146, 1088-1134, 11 18-1146, 1118-1143, 1127-1193, 1170-1189, 1176-1192, 1177-1192, 1203-1297, 1206-1255, 12091228, 1215-1234, 1218-1237, 1221- 1240, 1224-1243, 1227-1246, 1230-1249, 1233-1252, 1236-1255, 1251-1270, 1251-1285, 12541273, 1254-1279, 1257-1276, 1258-127 7, 1259-1278, 1260-1279, 1261-1285, 1261-1276, 1261-1277, 1261-1280, 1262-1281, 1262-1277, 1262-1278, 1263-1278, 1263-1279, 1263-1282, 1264-1297, 1264-1279, 1264-1280, 1264-1283, 1265-1280, 1265-1281, 1265-1284, 1266-1281, 1266-1282, 1266-1285, 126 7-1282, 1267-1283, 1268-1283, 1268-1284, 1268-1287, 1269-1284, 1269-1285, 12701285, 1281-1336, 1281-1324, 1281-1306, 1290-1 324, 1311-1336, 1326-1345, 1353-1381, 1395-1414, 1498-1535, 1498-1532, 1515-1535, 1515-1534, 1550-1655, 1553-1599, 1553-1590 1577-1606, 1577-1592, 1577-1593, 1577-1596, 1578-1593, 1578-1594, 15781597, 1579-1594, 1579-1595, 1579-1598, 1580-1595 580-1596, 1580-1599, 1581-1596, 1581-1597, 1581-1600, 1582-1597, 1582-1598, 1582-1601, 1583-1598, 1583-1599, 1583-1602, 1584 -1599, 1584-1600, 1584-1603, 1585-1600, 1585-1601, 1585-1604, 1586-1601, 1586-1602, 1586-1605, 1587-1602, 1587-1603, 1587-16 06, 1588-1603, 1588-1604, 1589-1604, 1589-1605, 1590-1605, 15901606, 1591-1606, 1642-1664, 1651-1720, 1655-1674, 1716-1738, 1743-1763, 1764-1783, 1773-1792, 1777-1800, 1777-1797, 1778-1800, 1778-1797, 1779-1799, 1778-1794, 1779-1794, 1779-1795, 17 79-1798, 1780-1796, 1780-1799, 1780-1795, 1781-1796, 17811797, 1781-1800, 1782-1797, 1794-1813, 1806-1837, 1806-1825, 1812- 1837, 1812-1831, 1815-1844, 1815-1834, 1818-1837, 18211837, 1822-1838, 1827-1846, 1861-1884, 1866-1885, 1867-1886, 1888-191 4, 1888-1907, 1891-1914, 1895-1938, 1895-1935, 1919-1938, 1928-1956, 1957-1976, 2035-2057, 2083-2141, 2230-2261, 2278-2297, 2281-2300, 2284-2303, 2368-2397, 2368-2396, 23682394, 2368-2393, 2379-2394, 2381-2396, 2420-2439, 2458-2476, 2819-2838, 287 3-2892, and 3161-3182.

[000269] In certain modalities, the following regions of nucleotide of seq ID no: 1, when they are targets of anti-Sensite or oligonucleotides compounds, display at least 70% of inhibition: 1-20, 10-29, 10-53, 13-38, 25-50, 43-68, 58-84, 58-74, 58-74, 58-75 , 59-80, 58-77, 60-75, 60-76, 61-77, 68-114, 98-123, 101-123, 113-138, 116-138, 131-150, 132-186, 191-215, 199-228, 199218, 200-223, 205-224, 206-228, 218-237, 224 -243, 233-263, 244-263, 245-264, 253-269, 253-274, 255-276, 256-279, 256-276, 256-274, 256272, 247-266, 250-265, 251-266, 253-272, 256-271, 266-291, 266-288, 260-279, 281-321, 281-303, 290-321, 290-312, 293-312, 302-321, 324343, 339-367, 339-361, 342-367, 358-392, 358-378, 360-392, 360-379, 366-39 2, 366-385, 370-392, 382-401, 405-428, 405-424, 409428, 411-433, 411-431, 411-430, 411-426, 412-431, 412-427, 413-428, 413-42 9, 413-432, 414-433, 414-430, 414-429, 415430, 414-433, 415-434, 415-435, 415-436, 416-434, 416-436, 416-432, 417-436, 417- 433, 418-433, 418-437, 423-436, 425465, 454-472, 455-472, 457-472, 457-476, 458-485, 458-483, 463-498, 467-498, 457-476, 470-493 470-493, 472-491, 485-519, 485513, 485-519, 485-513, 500-519, 512-534, 524-546, 536-558, 548-567, 554-573, 548-576, 560-594, 584-606, 608-648, 639-654, 640-656, 641656, 642-657, 642-658, 643-658, 653-672, 662-685, 665-685, 670-706, 670-689, 670-685, 671-690, 671-686, 671- 687, 672-687, 672688, 672-707, 672-697, 672-693, 673-688, 679-698, 681-696, 682-697, 682-701, 683-698, 684-699, 686-701, 687-702, 687-754, 687-702, 688703, 690-754, 690-706, 687-705, 687-703, 687-706, 692-711, 697-716, 724-758, 724-754, 724-752, 738-754, 739-754, 738- 754, 742785, 757-785, 790-815, 811-906, 811-844, 811-833, 822-867, 822-844, 845-867, 854-906, 854-873, 878-897, 899-958, 899-933, 936-958, 945964, 951-1044, 951-1024, 951-985, 951-997, 963-1044, 963-1024, 963-997, 966-985, 978-997, 1031-1056, 1046-1083, 1070-1095, 1081-1143, 1081-1134, 1082-1 101, 1088-1146, 1088-1134, 1118-1146, 1118-1143, 1127-1193, 1170-1189, 1176-1192, 1177-1192, 12031297, 1206-1255, 1209-1228, 1215-1234, 1218-1237, 1221-1240, 1224-1240 , 1227-1246, 1230-1249, 1233-1252, 1236-1255, 12511285, 1251-1270, 1254-1273, 1254-1279, 1257-1276, 1258-1277, 1259-1278, 1261-1285, 1261-1276, 1261-1277, 12611 280, 1262-1281, 1262-1277, 1262-1278, 1263-1278, 1263-1279, 1263-1282, 1264-1297, 1264-1279, 1264-1280, 1264-1283, 1265-1284, 1266-1282, 1266-1285, 1267-1282 , 1267-1283, 1268-1283, 1268-1284, 1268-1287, 1269-1284, 1269-1285, 12701285, 1281-1336, 1281-1306, 1290-1324, 1311-1336, 1353-1381, 1395-1414, 1498 -1535, 1498-1532, 15151535, 1550-1655, 1553-1599, 1553-1590, 1577-1606, 1577-1592, 1577-1593, 1578-1593, 1578-1594, 1578-1597, 1579-1595, 1579-1598, 1579-1598, 1580-1595, 1580-1596, 1580-1599, 1581-1596, 1581-1597, 1581-1600, 1582-1597, 1582-1598, 15821601, 1583-1598, 1583-1599, 1583-1602, 1584-1600, 1584-1603, 158-1600, 15 85-1601, 1585-1604, 1586-1602, 15861605, 1587-1602, 1587-1603, 1587-1606, 1588-1603, 1588-1604, 1589-1604, 1590-1605, 1591-1606, 1591-1606, 1651-1720, 1716-1738, 1743 -1763, 1764-1783, 1773-1792, 1777-1800, 1777-1797, 1778-1800, 1778-1797, 1779-1799, 17781794, 1779-1795, 1779-1798, 1780-1795, 1780-1796, 1780-1799, 1782-1797 94-1813, 1806-1837, 1806-1825, 18121837, 1812-1831, 1815-1844, 1815-1834, 1818-1837, 1821-1837, 1822-1838, 1827-1846, 1861-1884, 1867-1886, 1881914, 1888-1 907, 1891-1914, 1895-1938, 1895-1935, 1919-1938, 1928-1956, 1957-1976, 2035-2057, 2083-2141, 2230-2261, 22782297, 2281-2300, 2284-2303, 2368-2397, 2368-2396, 2368- 2394, 2368-2393, 2379-2394, 2381-2396, 2420-2439, 2458-2476, 2819 2838, 2873-2892 and 3161-3182.

[000270] In certain embodiments, the following nucleotide regions of SEQ ID NO: 1, when targeted by antisense compounds or oligonucleotides, exhibit at least 75% inhibition: 13-32, 16-35, 19-38, 25-44, 28-47, 31-50, 43-62, 46-65, 49-68, 55 1 40-159, 143-162, 158-177, 161-180, 164-183, 167-186, 200-219, 203-226, 209-228, 218-237, 233-252, 236-255, 239-258, 242-264 , 247-266, 251-266, 253-272, 255-276, 266-285, 269-288, 281-300, 284-303, 290-313, 298-317, 302321, 324-343, 339-358, 342-3 61, 348-367, 358-381, 364-383, 366-386, 370-389, 373-392, 382-401, 405-424, 409-428, 411-430, 411-426, 411427, 412-427, 412 -431, 413-428, 413-429, 413-432, 414-436, 414-430, 414-429, 415-430, 416-431, 416-432, 417-433, 418-437, 422-441, 425444, 4 28-447, 434-453, 440-459, 443-462, 446-465, 456-477, 458-473, 464-483, 470-493, 476-495, 479-498, 488-507, 491-510, 494-513 ,500519,512-531,515-534,524-543,527-546,536-555,539-558,560-579,566-585,569-588,572-591,575-594,584-603,587-6 06, 608-627, 614633, 617-636, 620-639, 623-642, 626-645, 629-648, 639-654, 641-656, 642-657, 643-658, 653-672, 665-684, 668 -688, 670-706, 670-686, 670685, 671-691, 671-687, 671-686, 672-688, 673-688, 679-703, 681-696, 682-697, 686-701, 686-706, 6 87-702, 687-703, 688-703, 689-708, 693712, 695-714, 696-715, 697-716, 727-746, 739-754, 742-761, 748-767, 751-770, 754-773, 757-776, 760-779, 763-782, 766-785, 790-809, 793812, 796-815, 811-830, 814-833, 817-836, 820-839, 822-844, 845-864, 854-87 3, 857-876, 863-882, 866-885, 872-891, 875-894, 878-897, 881900, 884-903, 887-906, 899-918, 902-921, 905-924, 908-927, 911- 930, 914-933, 936-955, 939-958, 951-970, 954-973, 957-976, 960-979, 963 982, 966-985, 969-988, 972-991, 975-994, 978-997, 9 96-1015, 10021021, 1025-1044, 1031-1050, 1034-1053, 1037-1056, 1046-1065, 1049-1068, 1052-1071, 1055-1074, 1058-1077, 1061- 1080, 10641083, 1070-1089, 1073-1092, 1076-1095, 1082-1101, 1088-1107, 1094-1113, 1097-1116, 1100-1119, 1103-1122, 1106-112 5, 11091128, 1112-1131, 1115-1134, 1121-1140, 1127-1146, 1153-1172, 1156-1175, 1159-1178, 1162-1181, 1165-1184, 1168-1191, 11741193, 1206-1225, 1209-1228, 1212-1231, 1215-1234, 1218-1237, 1221-1240, 1224-1243, 1227-1246, 1230-1249, 1233-1252, 123 61255, 1239-1258, 1242-1261, 1245-1264, 1251-1270, 1254-1273, 1254-1279, 1257-1283, 1257-1276, 1258-1277, 1260-1279, 126112 85, 1261-1276, 1261-1277, 1261-1280, 1262-1277, 1262-1278, 1263-1278, 1263-1279, 1263-1282, 1264-1279, 1264-1280, 12641283, 1265-1281, 1265-1284, 1266-1281, 1266-1282, 1266-1285, 1267-1282, 1267-1283, 1268-1283, 1268-1284, 1268-1287, 12691284, 12 69-1285, 1270-1285, 1272-1291, 1275-1294, 1282-1303, 1286-1306, 1290-1309, 1293-1312, 1296-1315, 1299-1318, 13051324, 1311- 1330, 1314-1333, 1317-1336, 1353-1381, 1356-1375, 1359-1378, 1498-1517, 1501-1520, 1504-1523, 1510-1529, 15531572, 1556-157 5, 1559-1578, 1562-1581, 1565-1584, 1571-1590, 1574-1599, 1577-1606, 1577-1592, 1577-1593, 1577-1596, 1578-1593, 1578-1594, 1578-1597, 1579-1594, 1579-1595, 1579-1598, 1580-1595, 1580-1596, 1581-1596, 1581-1597, 1581-1600, 1582-1597, 1582-1598, 158 2-1601, 1582-1602, 1583-1598, 1583-1599, 1583-1602, 1584-1599, 1584-1600, 1584-1603, 1585-1600, 1585-1601, 1585-1604, 1586 -1601, 1586-1605, 1586-1602, 1587-1602, 1587-1603, 1587-1606, 1588-1603, 1588-1604, 1589-1604, 1589-1605, 1590-1605, 1590-16 06, 1591-1606, 1604-1623, 1607-1626, 1630-1649, 1633-1652, 1645-1664, 1651-1670, 1654-1674, 16571676, 1660-1679, 1663-1682, 1666-1685, 1689-1708, 1695-1714, 1698-1717, 1701-1720, 1716-1735, 1778-1797, 1778-1794, 1778-1797, 1779-1795, 1779-1798, 17 80-1795, 1780-1796, 1780-1799, 1781-1800, 1794-1813, 1895-1914, 1898-1917, 1901-1920, 1907-1926, 1910-1929, 1913-1932, 1916- 1935, 1919-1938, 2278-2297, 2281-2300 and 2284-2303.

[000271] In certain embodiments, the following nucleotide regions of SEQ ID NO: 1, when targeted by antisense compounds or oligonucleotides, exhibit at least 80% inhibition: 13-32, 16-35, 19-38, 25-44, 28-47, 46-65, 49-68, 58-77, 59-80, 63 -82, 98-120, 116-135, 137-159, 158-177, 167-186, 203-224, 205-224, 209228, 218-237, 233-252, 236-263, 245-264, 253-272, 256 -275, 257-276, 266-288, 281-300, 290-312, 293-312, 324-343, 339-358, 348-367, 358-378, 360-379, 361-383, 366-385, 373-392, 382-401, 405-424, 411-431, 411-426, 411-427, 411-430, 413-428, 414-433, 414-434, 415-430, 415434, 416-431, 416-435, 417-436 ,418-437,422-441,425-444,434-453,456-476,458-473,458-477,464-483,471-493,488-507,494-513,512531,524-543,527-5 46, 536-558, 560-579, 566-585, 572-591, 575-594, 584-603, 587-606, 608-627, 614-633, 617-636, 620-639, 623-642, 626645, 629 -648, 639-654, 641-656, 642-657, 643-658, 665-688, 670-687, 670-686, 671-686, 671-687, 671-691, 673-688, 679-699, 682-697, 682706, 686-701, 687-702, 687-706, 687-703, 693-715, 727-746, 742-761, 748-767, 757-776, 766-785, 790-815, 814-833, 820-839 . 015, 978-997, 1002-1021, 1025-1044, 1031-1056, 1049-1074, 1061-1083, 1070-1089, 1082-1101, 1088-11107, 1094-1119, 1109-113 4, 1121-1140, 1127-1146, 1159-1187, 1171-1191, 1206-1228, 1209-1228, 1215-1255, 12151234, 1218-1237, 1221-1240, 1224-1243, 1227-1246, 1230-1249, 1233-1252, 1236-1255, 1245-1264, 1251-1279, 1251-1270, 12541273, 1254-1279, 1257-1276, 1258-1277, 125 9-1278, 1260-1279, 1261-1285, 1261-1276, 1261-1277, 1261-1280, 1262-1277, 12621278, 1262-1281, 1263-1278, 1263-1282, 1264-1 279, 1264-1283, 1265-1284, 1266-1285, 1267-1282, 1267-1283, 1268-1283, 12681284, 1269-1284, 1269-1285, 1269-1288, 1270-1285 . 577-1592, 1577-1596, 1577-1593, 1577-1596, 1578-1593, 1578-1594, 1578-1597, 1579-1595, 1579-1598, 1580-1596, 1580-1599, 1581 -1596, 1581-1597, 1581-1600, 1582-1597, 1582-1601, 1582-1602, 1583-1598, 1583-1599, 1583-1602, 1584-1599, 1584-1603, 1585-16 01, 1585-1604, 1586-1605, 1587-1602, 1587-1606, 1588-1603, 15891604, 1589-1605, 1657-1679, 1780-1795, 1780-1796, 1780-1799, 1913-1935, 2278-2297, 2281-2300 and 2284-2303.

[000272] In certain embodiments, the following nucleotide regions of SEQ ID NO: 1, when targeted by antisense compounds or oligonucleotides, exhibit at least 85% inhibition: 13-32, 16-35, 19-38, 25-44, 46-65, 59-80, 101-120, 140-159, 158-1 77, 167-186, 200-219, 205-224, 209-228, 233-252, 242-263, 253-272, 266285, 281-300, 290-311, 293-312, 359-379, 361-381, 370 -389, 382-401, 411-426, 411-430, 411-427, 413-428, 414-433, 415-430, 416-435, 417-436, 422-441, 456-476, 458-473, 470-493, 512-531, 524-543, 536-558, 566-585, 575-594, 587-606, 608-627, 614-636, 623-645, 639-654, 665687, 671-686, 671-687, 680-699 , 682-703, 687-706, 687-703, 727-746, 742-761, 757-776, 793-812, 822-843, 854-876, 854-873, 863-885, 878900, 878-897, 887-9 06 899-918 905-927 914-933 936-955 951-985 966-985 972-1015 978-997 1002-1021 1025-1044 1037-1056 1049-1074 10 64-1083, 1070-1089, 1088-1107, 1094-1119, 11091128, 1121-1140, 1156-1175, 1162-1187, 1172-1191, 1206-1228, 1209-1228, 1215- 1255, 1215-1234, 1218-1237, 1221-1240, 12241243, 1227-1246, 1230-1249, 1233-1252, 1236-1255, 1245-1264, 1251-1279, 1251-127 0. 1262-1278, 1262-1281, 1263-1278, 1263-1282, 1264-1279, 1264-1283, 1265-1284, 1266-1285, 1267-1282, 1267-1283, 1268-1284, 126 9-1284, 1269-1285, 1269-1288, 1270-1285, 1275-1294, 1282-1301, 1293-1315, 1311-1330, 1359-1378, 1574-1593, 1577-1592, 1577-1 593, 1577-1596, 1577-1606, 1578-1593, 1578-1594, 1578-1597, 1579-1598, 1580-1596, 1580-1599, 1581-1597, 1581-1600, 1582-1601 , 1583-1598, 1583-1602, 15841603, 1585-1601, 1585-1604, 1586-1605, 1587-1602, 1588-1603, 1780-1799, 1780-1796, and 2278-2297, 2281-2300 and 2284-2303.

[000273] In certain embodiments, the following nucleotide regions of SEQ ID NO: 1, when targeted by antisense compounds or oligonucleotides, exhibit at least 90% inhibition: 13-32, 16-35, 60-80, 140-159, 158-177, 167-186, 242-261, 292-311, 362-381, 370-389, 382-401, 411-427, 411-426, 413-428, 415-430, 416435, 422-441, 473-492, 617-636, 623-642, 639-654, 668-68 7, 680-699, 682-701, 684-703, 687-706, 727-746, 757-776, 824-843, 854-873, 854-876, 863-882, 878-897, 878-900, 887-906, 899 -918, 905-927, 914-933, 936-955, 951-970, 960-985, 966-985, 972-1015, 978-997, 1025-1044, 1037-1056, 1070-1089, 1097-1119, 1109-1128, 1121-1140, 1165-1187, 1172-1191, 1206-1228, 1209-1228, 1215-1234, 1215-1234, 1215-1255, 1218-1237, 1221-1240, 122 4-1243, 1227-1246, 1230-1249, 1233-1252, 1236-1255, 1245-1264, 1251-1279, 1251-1270, 1254-1273, 1254-1279, 1257-1276, 1258- 1277, 1259-1278, 1260-1279, 1261-1285, 1261-1280, 1262-1278, 1261-1276, 1262-1281, 1262-1277, 1263-1282, 1263-1278, 1264-1 283, 1265-1284, 1266-1285, 1268-1284, 1269-1284, 1269-1285, 1269-1288, 1296-1315, 1577-1605, 1577-1596, 1577-1593, 1577-1592 2 278-2297, 2281-2300 and 2284-2303.

[000274] In certain embodiments, the following nucleotide regions of SEQ ID NO: 1, when targeted by antisense compounds or oligonucleotides, exhibit at least 95% inhibition: 411-426, 411-427, 413-428, 617-636, 623-642, 668-687, 680-699, 68 2701, 854-873, 878-897, 887-906, 914-933, 966-985, 978-997, 12091228, 1215-1234, 1218-1237, 1221-1240, 1224-1243, 1227-1246 . 261-1285, 1261-1280, 1262-1281, 1263-1282, 1263-1278, 12641283, 1265-1284, 1266-1285, 1268-1284, 1269-1288, 1577-1592, 1577 -1596, 1577-1601, 1583-1598, 1585-1601, 1588-1603, 1780 1799, 2278-2297, 2281-2300 and 2284-2303.

[000275] In certain embodiments, the following antisense compounds or oligonucleotides target a region of an HBV nucleic acid and effect at least 50% inhibition of an HBV mRNA, ISIS IDs: 510088, 510089, 510090, 510092, 510096, 510097, 510098, 5100 99, 510100, 510101, 510102, 505330, 509928, 510104, 509929, 510105, 509930, 510106, 510107, 510108, 510111, 510115, 509931, 510116, 510117, 510118, 510119, 510120, 510121, 509932, 510122, 509933, 510123, 509934, 510124, 509935, 510125, 510126, 51 0127, 510128, 510140, 146779, 505314, 505315, 505316, 505317, 146821, 505318, 509922, 505319, 509925, 505320, 509952, 50532 1, 505322, 505323, 505324, 505325, 505326, 505327, 505328, 505329, 509956, 509957, 509927, 509958, 510038, 505330, 509959, 510039, 509960, 510040, 509961, 510041, 509962, 509963, 505331, 505332, 509968, 509969, 510050, 510052, 505333, 505334, 505 335, 505336, 509972, 146823, 509974, 505338, 505339, 509975, 505340, 509978, 505341, 509979, 510058, 505342, 509981, 510061 ,505344,505345,509983,505346,509984,505347,505348,505350,505352,505353,505354,505355,505356,146786,505357,5 05358, 505359, 505360, 509985, 509986, 509987, 509988, 505363, 505364, 505365, 505366, 146787, 510079, 524410, 524411, 5244 13, 524414, 524415, 524416, 524417, 524418, 524419, 524420, 524421, 524422, 524424, 524425, 524426, 524427, 524428, 524429, 524431, 524432, 524433, 524434, 524435, 524436, 524439, 524440, 524442, 524444, 524446, 524447, 524448, 524450, 524451, 52 4452, 524453, 524454, 524455, 524456, 524457, 524458, 524459, 524460, 524461, 524462, 524464, 524466, 524467, 524468, 52446 9, 524470, 524471, 524472, 524473, 524474, 524475, 524477, 524478, 524479, 524480, 524481, 524482, 524483, 524484, 524485, 524486, 524487, 524489, 524490, 524491, 524492, 524493, 524494, 524495, 524496, 524498, 524499, 524500, 524501, 524502, 524 503, 524504, 524506, 524507, 524508, 524509, 524510, 524511, 524512, 524513, 524514, 524515, 524516, 524517, 524518, 524519 524520 524521 524522 524523 524524 524525 524526 524527 524528 524529 524530 524531 524532 524533 524534 5 24535, 524536, 524537, 524538, 524539, 524540, 524541, 524543, 524544, 524546, 524547, 524548, 524549, 524550, 524551, 5245 52, 524553, 524554, 524555, 524556, 524557, 524558, 524559, 524560, 524561, 524562, 524563, 524564, 524565, 524568, 524569, 524570, 524571, 524572, 524573, 524574, 524575, 524576, 524577, 524578, 524579, 524580, 524581, 524582, 524584, 524585, 52 4586, 524587, 524588, 524589, 524590, 524591, 524592, 524593, 524594, 524595, 524598, 524599, 524600, 524601, 524602, 52460 3, 524604, 524605, 524606, 524607, 524608, 524609, 524610, 524611, 524614, 524615, 524616, 524617, 524618, 524619, 524620, 524621, 524622, 524623, 524624, 524625, 524626, 524627, 524629, 524632, 524633, 524634, 524635, 524636, 524637, 524638, 524 639, 524640, 524641, 524642, 524643, 524644, 524646, 524647, 524648, 524649, 524650, 524651, 524652, 524654, 524656, 524657 ,524658,524659,524660,524661,524662,524663,524664,524665,524666,524667,524668,524669,524670,524672,524673,5 24675, 524676, 524678, 524679, 524680, 524682, 524683, 524684, 524685, 524686, 524687, 524688, 524689, 524690, 524691, 5246 92, 524693, 524694, 524695, 524696, 524697, 524698, 524699, 524700, 524701, 524702, 524703, 524704, 524705, 524706, 524707, 524708, 524709, 524710, 524712, 524713, 524714, 524715, 524716, 524717, 524718, 524719, 524721, 524722, 524723, 524724, 52 4726, 524727, 524728, 524729, 524730, 524731, 524732, 524733, 524734, 524735, 524736, 524737, 524738, 524739, 524740, 52474 1, 524742, 524743, 524744, 524745, 524746, 524747, 524748, 524749, 524750, 524751, 524752, 524753, 524754, 524755, 524756, 524757, 524758, 524759, 524760, 524761, 524762, 524763, 524764, 524765, 524766, 524767, 524768, 524769, 524770, 524771, 524 772, 524773, 524774, 524775, 524776, 524777, 524778, 524779, 524780, 524781, 524782, 524783, 524784, 524785, 524786, 524787 ,524788,524789,524790,524791,524792,524793,524794,524795,524796,524797,524798,524799,524800,524801,524802,5 24803, 524804, 524805, 524806, 524807, 524808, 524809, 524810, 524811, 524812, 524813, 524814, 524815, 524816, 524817, 5248 18, 524819, 524820, 524821, 524822, 524823, 524824, 524825, 524826, 524827, 524828, 524829, 524830, 524831, 524632, 524833, 524834, 524835, 524842, 524843, 524844, 524845, 524847, 524848, 524856, 524857, 524861, 524866, 524867, 524868, 524869, 52 4870, 524871, 524872, 524873, 524875, 524876, 524877, 524878, 524879, 524880, 524881, 524882, 524883, 524884, 524885, 52488 6, 524887, 524888, 524889, 524890, 524891, 524892, 524893, 524894, 524895, 524896, 524897, 524898, 524899, 524900, 524901, 524902, 524903, 524904, 524905, 524906, 524907, 524908, 524909, 524910, 524911, 524912, 524913, 524914, 524915, 524916, 524 917, 524918, 524919, 524921, 524922, 524923, 524924, 524925, 524926, 524927, 524928, 524929, 524930, 524931, 524932, 524933 ,524934,524935,524936,524937,524938,524939,524940,524941,524942,524943,524944,524945,524946,524947,524948,5 24949, 524950, 524951, 524952, 524953, 524954, 524955, 524956, 524957, 524958, 524959, 524960, 524961, 524962, 524964, 5249 65, 524976, 524977, 524978, 524979, 524980, 524981, 524982, 524983, 524984, 524985, 524986, 524987, 524988, 524989, 524991, 524992, 524993, 524994, 524997, 524998, 525021, 525022, 525037, 525039, 525043, 525050, 525052, 525086, 525090, 525100, 55 1909, 551910, 551911, 551912, 551913, 551916, 551917, 551918, 551919, 551920, 551921, 551922, 551923, 551924, 551925, 55192 6, 551927, 551928, 551929, 551930, 551932, 551933, 551934, 551935, 551936, 551937, 551939, 551940, 551941, 551942, 551943, 551944, 551945, 551946, 551947, 551948, 551949, 551950, 551951, 551952, 551953, 551954, 551955, 551956, 551957, 551958, 551 959, 551960, 551962, 551963, 551964, 551965, 551966, 551967, 551968, 551971, 551972, 551973, 551974, 551975, 551976, 551977 ,551978,551979,551980,551981,551982,551983,551984,551985,551986,551987,551988,551989,551990,551992,551993,5 51994, 551995, 551996, 551997, 551998, 551999, 552000, 552001, 552002, 552003, 552004, 552005, 552006, 552007, 552009, 5520 10, 552011, 552012, 552013, 552014, 552015, 552016, 552017, 552018, 552019, 552020, 552021, 552022, 552023, 552024, 552025, 552026, 552027, 552028, 552029, 552030, 552031, 552032, 552033, 552034, 552035, 552036, 552037, 552038, 552039, 552040, 55 2041, 552042, 552043, 552044, 552045, 552046, 552047, 552048, 552049, 552050, 552051, 552052, 552053, 552054, 552055, 55205 6, 552057, 552058, 552059, 552060, 552061, 552062, 552063, 552064, 552065, 552067, 552068, 552069, 552070, 552071, 552072, 552073, 552074, 552075, 552076, 552077, 552078, 552079, 552080, 552081, 552082, 552083, 552084, 552085, 552086, 552087, 552 088, 552089, 552090, 552091, 552092, 552093, 552094, 552095, 552096, 552097, 552098, 552099, 552100, 552101, 552102, 552114 ,552115,552116,552117,552118,552119,552122,552123,552124,552125,552126,552127,552128,552129,552131,552132,5 52133, 552134, 552135, 552136, 552137, 552138, 552139, 552140, 552141, 552142, 552143, 552144, 552145, 552146, 552147, 5521 48, 552149, 552150, 552151, 552152, 552153, 552154, 552155, 552158, 552159, 552160, 552161, 552162, 552163, 552164, 552165, 552167, 552168, 552169, 552170, 552171, 552175, 552176, 552177, 552178, 552179, 552180, 552181, 552182, 552183, 552185, 55 2186, 552187, 552188, 552189, 552191, 552192, 552193, 552194, 552195, 552196, 552197, 552198, 552199, 552200, 552201, 55220 2, 552203, 552204, 552205, 552206, 552207, 552208, 552209, 552210, 552211, 552212, 552213, 552214, 552215, 552216, 552217, 552218, 552220, 552222, 552224, 552225, 552230, 552239, 552240, 552241, 552242, 552243, 552246, 552247, 552248, 552249, 552 250, 552251, 552252, 552253, 552254, 552255, 552256, 552257, 552258, 552259, 552260, 552261, 552262, 552263, 552264, 552265 ,552266,552267,552268,552269,552270,552271,552279,552285,552288,552293,552294,552295,552296,552297,552300,5 52301, 552302, 552303, 552304, 552305, 552306, 552307, 552308, 552309, 552310, 552312, 552313, 552314, 552315, 552316, 5523 17, 552318, 552319, 552320, 552321, 552322, 552323, 552325, 552326, 552330, 552331, 552332, 552333, 552337, 552338, 552339, 552340, 552341, 552342, 552343, 552344, 552345, 552347, 552348, 552349, 552350, 552351, 552352, 552354, 552355, 552356, 55 2357, 552358, 552359, 552360, 552361, 552362, 552363, 552364, 552365, 552366, 552367, 552368, 552369, 552370, 552371, 55237 2, 552373, 552374, 552375, 552376, 552377, 552378, 552379, 552380, 552385, 552386, 552390, 552391, 552393, 552394, 552395, 552396, 552397, 552398, 552399, 552400, 552401, 552402, 552403, 552408, 552409, 552410, 552411, 552412, 552413, 552414, 552 415, 552416, 552417, 552418, 552419, 552420, 552421, 552422, 552423, 552424, 552425, 552428, 552430, 552431, 552432, 552433 552440 552442 552443 552444 552445 552446 552447 552448 552449 552450 552452 552453 552455 552456 552458 5 52459, 552464, 552465, 552466, 552467, 552468, 552469, 552470, 552471, 552472, 552473, 552474, 552475, 552476, 552477, 5524 78, 552479, 552480, 552481, 552482, 552484, 552485, 552486, 552487, 552488, 552490, 552491, 552493, 552497, 552499, 552500, 552501, 552502, 552503, 552504, 552505, 552506, 552508, 552509, 552510, 552511, 552512, 552513, 552514, 552515, 552516, 55 2517, 552520, 552521, 552522, 552523, 552525, 552526, 552527, 552528, 552529, 552530, 552531, 552532, 552533, 552534, 55253 5, 552538, 552539, 552540, 552541, 552542, 552544, 552547, 552548, 552553, 552554, 552555, 552557, 552558, 552559, 552561, 552562, 552565, 552566, 552567, 552568, 552569, 552570, 552571, 552572, 552576, 552577, 552578, 552579, 552580, 552581, 552 582, 552583, 552584, 552585, 552586, 552587, 552588, 552589, 552590, 552591, 552592, 552594, 552595, 552596, 552597, 552598 552600 552606 552608 552787 552788 552789 552790 552791 552794 552795 552796 552797 552798 552799 552800 5 52801, 552802, 552803, 552804, 552805, 552806, 552807, 552808, 552809, 552810, 552811, 552812, 552813, 552814, 552815, 5528 16, 552817, 552818, 552819, 552820, 552821, 552822, 552823, 552824, 552825, 552826, 552827, 552828, 552829, 552830, 552831, 552832, 552833, 552834, 552835, 552836, 552837, 552838, 552839, 552840, 552841, 552842, 552843, 552844, 552845, 552846, 55 2847, 552848, 552849, 552850, 552851, 552852, 552853, 552854, 552855, 552856, 552857, 552858, 552859, 552860, 552861, 55286 2, 552863, 552864, 552865, 552866, 552868, 552870, 552871, 552872, 552876, 552889, 552890, 552891, 552892, 552893, 552894, 552895, 552896, 552898, 552899, 552901, 552902, 552903, 552904, 552905, 552907, 552908, 552909, 552910, 552911, 552912, 552 913, 552914, 552915, 552916, 552917, 552918, 552919, 552922, 552923, 552925, 552926, 552927, 552928, 552929, 552930, 552931 552932 552933 552934 552935 552936 552937 552938 552939 552940 552941 552942 552943 552944 552945 552946 5 52947, 552948, 552950, 552951, 552953, 552954, 552955, 552956, 552957, 552958, 552959, 552960, 552961, 552965, 552966, 5529 69, 552970, 552971, 552972, 552973, 552974, 552975, 552976, 552977, 552979, 552980, 552981, 552982, 552983, 552984, 552987, 552988, 552989, 552990, 552991, 552992, 552993, 552994, 552995, 552996, 552997, 552998, 552999, 553000, 553001, 553002, 55 3003, 553004, 553005, 553006, 553007, 553008, 553009, 553010, 553011, 553012, 553014, 553015, 553016, 566828, 566829, 56683 0, 566831, 566832, 577120, 577121, 577122, 577123, 577124, 577125, 577126, 577127, 577128, 577129, 577130, 577131, 577132, 577133, 577134, 577135, 577136, 582665 and 582666.

[000276] In certain embodiments, the following antisense compounds or oligonucleotides target a region of an HBV nucleic acid and effect at least 50% inhibition of an HBV mRNA, SEQ ID NOs: 5, 6, 7, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 33, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 74, 83, 85 ,86,87,88,89,92,96,98,99,100,102,103,104,106,108,109,111,112,115,117,121,122,123,124,125,126,127, 128, 136, 137, 139, 140, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 153, 155, 157, 159, 161, 165, 166, 167, 168, 16 9, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 186, 187, 188, 189, 190, 191, 192, 193, 194, 197, 198, 199, 201, 203, 206, 207, 208, 209, 210, 211, 212, 213, 215, 217, 218, 220, 221, 222, 224, 225, 226, 227, 228, 230, 231, 232, 233, 234 ,235,236,237,240,241,242,243,244,250,283,321,322,324,325,326,327,328,329,330,331,332,333,335,336,3 37, 338, 339, 340, 342, 343, 344, 345, 346, 347, 350, 351, 353, 355, 357, 358, 359, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384, 387, 388, 389, 390, 391, 392, 393, 394, 395, 39 6, 397, 399, 400, 401, 402, 403, 404, 405, 406, 408, 409, 410, 411, 412, 413, 414, 416, 417, 418, 419, 420 421, 422, 423, 4 24, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 453, 454, 456, 457, 458, 459, 460, 461, 462, 463, 464, 465, 466, 467, 468, 469, 470, 471, 473, 474, 475, 47 8, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 494, 495, 496, 497, 498, 499, 500, 501, 502, 503, 504, 505, 508, 509, 510, 511, 512, 513, 514, 515, 516, 517, 518, 519, 520, 521, 524, 525, 526, 527, 528, 529, 530, 531, 532 ,533,534,535,536,537,539,542,543,544,545,546,547,548,549,550,551,552,553,554,555,556,557,558,559,5 60, 561, 562, 564, 566, 567, 568, 569, 570, 571, 572, 573, 574, 575, 576, 577, 578, 579, 580, 582, 583, 585, 586, 588, 589, 590, 592, 593, 594, 595, 596, 597, 598, 599, 600, 601, 602, 603, 604, 605, 606, 607, 608, 609, 610, 611, 612, 613, 614, 61 5, 616, 617, 618, 619, 620, 622, 623, 624, 625, 626, 627, 628, 629, 631, 632, 633, 634, 636, 637, 638, 639, 640, 641, 642, 643, 644, 645, 646, 647, 648, 649, 650, 651, 652, 653, 654, 655, 656, 657, 658, 659, 660, 661, 662, 663, 664, 665, 666, 667 ,668,669,670,671,672,673,674,675,676,677,678,679,680,681,682,683,684,685,686,687,688,689,690,691,6 92, 693, 694, 695, 696, 697, 698, 699, 700, 701, 702, 703, 704, 705, 706, 707, 708, 709, 710, 711, 712, 713, 714, 715, 716, 717, 718, 719, 720, 721, 722, 723, 724, 725, 726, 727, 728, 729, 730, 731, 732, 733, 734, 735, 736, 737, 738, 739, 740, 74 1, 742, 743, 744, 745, 746, 747, 754, 755, 756, 757, 759, 760, 768, 769, 773, 777, 778, 779, 780, 781, 782, 783, 784, 786, 787, 788, 789, 790, 791, 792, 793, 794, 795, 796, 797, 798, 799, 800, 801, 802, 803, 804, 805, 806, 807, 808, 809, 810, 811 ,812,813,814,815,816,817,818,819,820,821,822,823,824,825,826,827,828,829,830,831,833,834,835,836,8 37, 838, 839, 840, 841, 842, 843, 844, 845, 846, 847, 848, 849, 850, 851, 852, 853, 854, 855, 856, 857, 858, 859, 860, 861, 862, 863, 864, 865, 866, 867, 868, 869, 870, 871, 872, 873, 874, 876, 877, 888, 889, 890, 891, 892, 893, 894, 895, 896, 89 7, 898, 899, 900, 901, 903, 904, 905, 906, 909, 910, 933, 934, 949, 951, 955, 962, 964, 998, 1002, 1013, 1052, 1267, 1271, 1272, 1288, 1289, 1290, 1291, 1292, 1293, 1294, 1295, 1296, 1297, 1298, 1299, 1300, 1301, 1302, 1303, 1304, 1305, 1306, 130 7, 1308, 1309, 1310, 1311, 1312, 1313, 1314, 1315, 1316, 1317, 1318, 1319, 1320, 1321, 1322, 1323, 1324, 1325, 1326, 1327, 1328, 1329, 1330, 1331, 1332, 1333, 1334, 1335, 1336, 1337, 1338, 1339, 1340, 1341, 1342, 1343, 1344, 1345, 1346, 1347, 134 8, 1349, 1350, 1364, 1365, 1366, 1367, 1368, 1369, 1370, 1371, 1372, 1375 and 1376.

[000277] In certain embodiments, the following antisense compounds or oligonucleotides target a region of an HBV nucleic acid and effect at least 60% inhibition of an HBV mRNA, ISIS IDs: 510090, 510100, 510102, 505330, 509928, 510104, 509929, 5101 05, 509930, 510106, 510107, 510111, 509931, 510116, 510117, 510118, 510119, 510120, 510121, 509932, 510122, 509933, 510123, 509934, 510124, 509935, 510125, 510128, 146779, 505314, 505315, 505316, 505317, 146821, 505318, 505319, 505322, 505323, 50 5324, 505325, 505326, 505327, 505328, 505329, 509956, 509957, 509958, 505330, 509959, 510041, 505332, 509968, 505333, 50533 5, 146823, 509974, 505338, 505339, 509975, 505340, 505341, 509979, 505342, 509981, 505344, 505345, 509983, 505346, 509984, 505347, 505348, 505353, 505354, 505356, 146786, 505357, 505358, 505359, 505360, 509985, 509986, 505363, 505366, 524410, 524 413, 524414, 524415, 524416, 524417, 524418, 524419, 524420, 524421, 524422, 524424, 524425, 524426, 524428, 524431, 524432 ,524433,524434,524435,524439,524440,524446,524447,524448,524451,524452,524453,524454,524455,524456,524457,5 24459, 524460, 524461, 524464, 524466, 524467, 524468, 524469, 524471, 524472, 524473, 524474, 524475, 524477, 524478, 5244 79, 524480, 524481, 524482, 524485, 524486, 524487, 524489, 524490, 524491, 524492, 524493, 524494, 524495, 524496, 524499, 524500, 524501, 524502, 524503, 524504, 524506, 524507, 524508, 524509, 524510, 524511, 524512, 524513, 524514, 524515, 52 4516, 524517, 524519, 524520, 524521, 524523, 524525, 524526, 524527, 524528, 524529, 524532, 524533, 524534, 524535, 52453 6, 524537, 524538, 524539, 524540, 524541, 524543, 524546, 524547, 524549, 524550, 524552, 524553, 524554, 524555, 524556, 524557, 524558, 524559, 524560, 524561, 524562, 524563, 524564, 524565, 524568, 524569, 524570, 524571, 524572, 524573, 524 574, 524575, 524576, 524577, 524578, 524579, 524580, 524581, 524582, 524585, 524586, 524587, 524588, 524589, 524590, 524591 ,524593,524594,524595,524598,524599,524600,524602,524603,524604,524605,524606,524607,524610,524611,524614,5 24615, 524616, 524617, 524618, 524619, 524620, 524621, 524622, 524623, 524625, 524627, 524629, 524632, 524633, 524634, 5246 35, 524636, 524637, 524638, 524639, 524640, 524641, 524642, 524643, 524644, 524646, 524647, 524648, 524649, 524650, 524651, 524654, 524656, 524657, 524658, 524659, 524661, 524662, 524663, 524664, 524665, 524666, 524667, 524668, 524669, 524670, 52 4673, 524675, 524676, 524678, 524679, 524680, 524683, 524684, 524685, 524686, 524687, 524688, 524689, 524690, 524691, 52469 2, 524694, 524695, 524696, 524697, 524698, 524699, 524700, 524701, 524702, 524703, 524704, 524705, 524706, 524707, 524708, 524709, 524710, 524713, 524714, 524715, 524716, 524717, 524718, 524719, 524721, 524722, 524724, 524726, 524727, 524728, 524 729, 524730, 524731, 524732, 524733, 524734, 524735, 524736, 524737, 524738, 524739, 524741, 524742, 524743, 524744, 524746 ,524747,524748,524749,524750,524751,524752,524753,524754,524755,524756,524757,524758,524759,524760,524761,5 24762, 524763, 524764, 524765, 524766, 524767, 524768, 524769, 524770, 524771, 524772, 524773, 524774, 524775, 524776, 5247 77, 524778, 524779, 524780, 524781, 524782, 524783, 524784, 524785, 524787, 524788, 524789, 524790, 524791, 524792, 524793, 524794, 524795, 524796, 524797, 524798, 524799, 524800, 524801, 524802, 524803, 524804, 524805, 524806, 524807, 524808, 52 4809, 524810, 524811, 524812, 524813, 524814, 524815, 524816, 524817, 524818, 524819, 524820, 524821, 524822, 524823, 52482 4, 524825, 524826, 524827, 524828, 524829, 524830, 524632, 524833, 524842, 524843, 524844, 524845, 524847, 524856, 524866, 524867, 524868, 524869, 524870, 524871, 524872, 524873, 524876, 524878, 524879, 524880, 524881, 524882, 524883, 524884, 524 885, 524886, 524887, 524888, 524889, 524890, 524891, 524892, 524893, 524894, 524895, 524896, 524897, 524898, 524899, 524900 ,524901,524902,524903,524904,524905,524906,524907,524908,524909,524910,524911,524912,524913,524914,524915,5 24916, 524921, 524922, 524923, 524924, 524925, 524926, 524928, 524929, 524930, 524931, 524932, 524933, 524936, 524937, 5249 38, 524939, 524940, 524941, 524942, 524944, 524946, 524947, 524948, 524949, 524950, 524952, 524953, 524954, 524955, 524961, 524977, 524978, 524979, 524980, 524981, 524982, 524983, 524984, 524985, 524986, 524987, 524988, 524991, 524992, 524993, 52 4994, 525037, 525052, 551909, 551911, 551919, 551920, 551921, 551922, 551924, 551925, 551926, 551927, 551928, 551932, 55193 3, 551934, 551935, 551936, 551941, 551943, 551944, 551948, 551949, 551950, 551951, 551952, 551953, 551954, 551955, 551956, 551957, 551958, 551959, 551960, 551962, 551963, 551965, 551966, 551967, 551968, 551973, 551975, 551979, 551981, 551982, 551 983, 551984, 551985, 551986, 551987, 551989, 551990, 551992, 551993, 551994, 551995, 551996, 551997, 551998, 551999, 552000 552001, 552002, 552003, 552005, 552006, 552007, 552009, 552010, 552012, 552013, 552014, 552015, 552016, 552017, 552018, 5 52019, 552020, 552021, 552022, 552023, 552024, 552025, 552026, 552027, 552028, 552029, 552030, 552031, 552032, 552033, 5520 34, 552035, 552036, 552038, 552039, 552041, 552042, 552044, 552045, 552046, 552047, 552048, 552049, 552050, 552051, 552052, 552053, 552054, 552055, 552056, 552057, 552058, 552059, 552060, 552061, 552062, 552063, 552064, 552065, 552068, 552069, 55 2070, 552071, 552073, 552074, 552075, 552076, 552077, 552078, 552079, 552080, 552081, 552082, 552083, 552084, 552085, 55208 6, 552087, 552088, 552089, 552090, 552091, 552092, 552093, 552094, 552095, 552096, 552097, 552098, 552099, 552100, 552101, 552102, 552114, 552115, 552116, 552117, 552118, 552119, 552123, 552124, 552125, 552126, 552127, 552128, 552129, 552131, 552 132, 552133, 552134, 552135, 552136, 552138, 552139, 552140, 552141, 552143, 552144, 552145, 552146, 552147, 552148, 552149 552150 552151 552152 552153 552155 552158 552159 552160 552162 552163 552168 552169 552170 552171 552176 5 52178, 552179, 552180, 552182, 552183, 552185, 552187, 552188, 552191, 552192, 552193, 552194, 552195, 552196, 552197, 5521 98, 552199, 552200, 552201, 552202, 552203, 552204, 552205, 552206, 552207, 552208, 552209, 552210, 552211, 552212, 552213, 552214, 552215, 552216, 552222, 552224, 552225, 552239, 552240, 552242, 552246, 552247, 552248, 552252, 552253, 552254, 55 2255, 552256, 552257, 552258, 552259, 552261, 552263, 552265, 552266, 552268, 552285, 552293, 552294, 552295, 552296, 55230 1, 552302, 552303, 552306, 552307, 552308, 552309, 552310, 552312, 552313, 552314, 552315, 552316, 552317, 552318, 552320, 552321, 552322, 552323, 552325, 552326, 552331, 552332, 552337, 552338, 552339, 552340, 552343, 552345, 552347, 552348, 552 349, 552351, 552354, 552355, 552356, 552358, 552359, 552360, 552361, 552362, 552363, 552364, 552365, 552366, 552367, 552368 552369 552370 552371 552372 552373 552374 552375 552376 552377 552378 552379 552396 552397 552398 552403 5 52408, 552409, 552410, 552411, 552412, 552414, 552416, 552418, 552419, 552420, 552421, 552422, 552423, 552424, 552431, 5524 42, 552445, 552449, 552455, 552456, 552459, 552464, 552465, 552466, 552467, 552469, 552472, 552473, 552474, 552475, 552477, 552478, 552479, 552480, 552484, 552487, 552497, 552508, 552509, 552511, 552512, 552515, 552516, 552520, 552521, 552522, 55 2523, 552526, 552527, 552528, 552529, 552530, 552531, 552534, 552540, 552541, 552542, 552559, 552567, 552568, 552569, 55257 0, 552572, 552576, 552577, 552578, 552579, 552582, 552583, 552584, 552585, 552586, 552587, 552588, 552590, 552595, 552596, 552597, 552788, 552789, 552790, 552791, 552796, 552800, 552801, 552803, 552804, 552805, 552806, 552807, 552808, 552809, 552 811, 552812, 552813, 552814, 552815, 552816, 552817, 552818, 552819, 552820, 552821, 552822, 552823, 552824, 552826, 552827 ,552828,552829,552830,552831,552832,552833,552834,552835,552836,552837,552838,552839,552841,552842,552843,5 52844, 552845, 552846, 552847, 552848, 552849, 552850, 552851, 552852, 552853, 552854, 552855, 552856, 552857, 552858, 5528 59, 552860, 552861, 552862, 552863, 552864, 552865, 552866, 552872, 552891, 552892, 552893, 552894, 552902, 552903, 552904, 552905, 552907, 552908, 552909, 552910, 552911, 552912, 552913, 552914, 552915, 552916, 552917, 552918, 552922, 552923, 55 2925, 552927, 552928, 552929, 552930, 552931, 552932, 552933, 552934, 552935, 552936, 552937, 552938, 552939, 552940, 55294 1, 552942, 552943, 552944, 552945, 552946, 552951, 552955, 552956, 552957, 552958, 552960, 552961, 552966, 552969, 552971, 552972, 552973, 552974, 552975, 552976, 552977, 552979, 552980, 552981, 552982, 552983, 552984, 552988, 552989, 552990, 552 991, 552992, 552993, 552994, 552995, 552996, 552998, 552999, 553000, 553001, 553002, 553003, 553004, 553005, 553006, 553007 553008 553009 553010 553011 553012 553016 566828 566829 566830 566831 566832 577120 577121 577122 577123 5 77124, 577125, 577126, 577127, 577128, 577129, 577130, 577131, 577132, 577133, 577134, 577135, 577136 and 582666.

[000278] In certain embodiments, the following antisense compounds or oligonucleotides target a region of an HBV nucleic acid and effect at least 60% inhibition of an HBV mRNA, SEQ ID NOs: 7, 9, 10, 12, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 2 8, 29, 33, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 56, 83, 85, 86, 87, 88, 89, 92, 96, 98, 100, 102, 10 3, 112, 115, 117, 122, 123, 124, 125, 126, 127, 128, 136, 137, 139, 140, 142, 143, 145, 147, 149, 150, 151, 153, 155, 157, 159, 161, 166, 167, 168, 172, 174, 176, 177, 178, 179, 180, 181, 186, 187, 188, 189, 190, 191, 192, 193, 194, 198, 199, 201 ,206,207,208,209,210,211,212,213,218,220,222,224,225,226,227,228,230,231,232,233,234,240,243,321,3 24, 325, 326, 327, 328, 329, 330, 331, 332, 333, 335, 336, 337, 339, 342, 343, 344, 345, 346, 350, 351, 357, 358, 359, 362, 363, 364, 365, 366, 367, 368, 370, 371, 372, 375, 376, 377, 378, 379, 381, 382, 383, 384, 387, 388, 389, 390, 391, 392, 39 5, 396, 397, 399, 400, 401, 402, 403, 404, 405, 406, 409, 410, 411, 412, 413, 414, 416, 417, 418, 419, 420 421, 422, 423, 4 24, 425, 426, 427, 429, 430, 431, 433, 435, 436, 437, 438, 439, 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 453, 456, 457, 459, 460, 462, 463, 464, 465, 466, 467, 468, 469, 470, 471, 473, 474, 475, 478, 479, 480, 481, 482, 483, 484, 485, 48 6, 487, 488, 489, 490, 491, 492, 495, 496, 497, 498, 499, 500, 501, 503, 504, 505, 508, 509, 510, 512, 513, 514, 515, 516, 517, 520, 521, 524, 525, 526, 527, 528, 529, 530, 531, 532, 533, 535, 537, 539, 542, 543, 544, 545, 546, 547, 548, 549, 550 ,551,552,553,554,555,556,557,558,559,560,561,564,566,567,568,569,571,572,573,574,575,576,577,578,5 79, 580, 583, 585, 586, 588, 589, 590, 593, 594, 595, 596, 597, 598, 599, 600, 601, 602, 604, 605, 606, 607, 608, 609, 610, 611, 612, 613, 614, 615, 616, 617, 618, 619, 620, 623, 624, 625, 626, 627, 628, 629, 631, 632, 634, 636, 637, 638, 639, 64 0, 641, 642, 643, 644, 645, 646, 647, 648, 649, 650, 652, 653, 654, 655, 657, 658, 659, 660, 661, 662, 663, 664, 665, 666, 667, 668, 669, 670, 671, 672, 673, 674, 675, 676, 677, 678, 679, 680, 681, 682, 683, 684, 685, 686, 687, 688, 689, 690, 691 ,692,693,694,695,696,698,699,700,701,702,703,704,705,706,707,708,709,710,711,712,713,714,715,716,7 17, 718, 719, 720, 721, 722, 723, 724, 725, 726, 727, 728, 729, 730, 731, 732, 733, 734, 735, 736, 737, 738, 740, 741, 742, 744, 745, 754, 755, 756, 757, 759, 768, 777, 778, 779, 780, 781, 782, 783, 784, 787, 789, 790, 791, 792, 793, 794, 795, 79 6, 797, 798, 799, 800, 801, 802, 803, 804, 805, 806, 807, 808, 809, 810, 811, 812, 813, 814, 815, 816, 817, 818, 819, 820, 821, 822, 823, 824, 825, 826, 827, 828, 833, 834, 835, 836, 837, 838, 840, 841, 842, 843, 844, 845, 848, 849, 850, 851, 852 ,853,854,856,858,859,860,861,862,864,865,866,867,873,889,890,891,892,893,894,895,896,897,898,899,9 00, 903, 904, 905, 906, 949, 964, 1271, 1288, 1289, 1290, 1291, 1292, 1293, 1294, 1295, 1296, 1297, 1298, 1299, 1300, 1301, 1302, 1303, 1304, 1305, 1306, 1307, 1308, 1309, 1310, 1311, 1312, 1313, 1314, 1315, 1316, 1317, 1318, 1319, 1320, 1321, 13 22, 1323, 1324, 1325, 1326, 1327, 1328, 1329, 1330, 1331, 1332, 1333, 1334, 1335, 1336, 1337, 1338, 1339, 1340, 1341, 1342, 1343, 1344, 1345, 1346, 1347, 1348, 1349, 1350, 1365, 1366, 1367, 1368, 1369, 1370, 1371, 1372 and 1376.

[000279] In certain embodiments, the following antisense compounds or oligonucleotides target a region of an HBV nucleic acid and effect at least 70% inhibition of an HBV mRNA, ISIS IDs: 510100, 510106, 509931, 510116, 510119, 509933, 510123, 5099 34, 510124, 505318, 505319, 505323, 505325, 505332, 505335, 509974, 505338, 505345, 505346, 505347, 505348, 505363, 524410, 524413, 524414, 524420, 524421, 524424, 524425, 524433, 524434, 524435, 524446, 524457, 524459, 524460, 524461, 524469, 52 4472, 524473, 524474, 524480, 524481, 524482, 524485, 524493, 524494, 524495, 524499, 524508, 524510, 524511, 524512, 52451 7, 524520, 524525, 524526, 524535, 524536, 524537, 524538, 524549, 524552, 524553, 524554, 524559, 524560, 524561, 524563, 524570, 524571, 524572, 524573, 524579, 524580, 524582, 524586, 524595, 524598, 524600, 524602, 524607, 524610, 524611, 524 614, 524619, 524620, 524621, 524629, 524637, 524638, 524641, 524642, 505330, 509928, 509929, 509930, 510120, 510121, 509932 ,510122,509935,146779,505317,146821,505326,505327,509957,505330,505339,509975,505342,509981,146786,505357,5 05358, 505359, 524415, 524416, 524418, 524419, 524426, 524428, 524431, 524432, 524447, 524448, 524452, 524453, 524464, 5244 66, 524467, 524468, 524475, 524477, 524478, 524479, 524487, 524490, 524491, 524492, 524500, 524502, 524503, 524507, 524513, 524514, 524515, 524516, 524528, 524532, 524533, 524534, 524539, 524540, 524541, 524547, 524555, 524556, 524557, 524558, 52 4564, 524565, 524568, 524569, 524574, 524575, 524577, 524578, 524587, 524590, 524591, 524594, 524603, 524604, 524605, 52460 6, 524615, 524616, 524617, 524618, 524633, 524634, 524635, 524636, 524643, 524644, 524646, 524647, 524648, 524649, 524650, 524651, 524656, 524657, 524659, 524661, 524662, 524663, 524664, 524665, 524666, 524667, 524668, 524669, 524670, 524678, 524 679, 524680, 524685, 524686, 524687, 524688, 524689, 524690, 524691, 524692, 524695, 524696, 524698, 524699, 524700, 524701 ,524702,524703,524704,524705,524706,524707,524708,524709,524713,524714,524715,524716,524717,524718,524721,5 24722, 524724, 524726, 524727, 524728, 524729, 524730, 524731, 524732, 524733, 524734, 524735, 524736, 524737, 524738, 5247 39, 524741, 524742, 524743, 524746, 524747, 524748, 524749, 524750, 524751, 524752, 524754, 524755, 524756, 524758, 524760, 524761, 524762, 524763, 524764, 524765, 524766, 524767, 524768, 524769, 524771, 524773, 524775, 524776, 524777, 524778, 52 4779, 524780, 524781, 524782, 524783, 524784, 524785, 524787, 524788, 524789, 524790, 524791, 524792, 524793, 524794, 52479 5, 524796, 524797, 524798, 524799, 524800, 524801, 524802, 524803, 524804, 524805, 524806, 524807, 524808, 524809, 524810, 524811, 524812, 524813, 524814, 524815, 524816, 524817, 524818, 524819, 524821, 524822, 524823, 524824, 524825, 524826, 524 827, 524828, 524829, 524830, 524833, 524842, 524843, 524844, 524845, 524856, 524866, 524867, 524868, 524869, 524870, 524871 ,524873,524879,524880,524881,524882,524883,524884,524885,524886,524887,524888,524889,524890,524891,524892,5 24893, 524894, 524895, 524896, 524897, 524898, 524899, 524900, 524902, 524903, 524905, 524906, 524907, 524908, 524909, 5249 10, 524911, 524912, 524913, 524914, 524915, 524916, 524921, 524922, 524930, 524931, 524932, 524937, 524940, 524942, 524948, 524980, 524981, 524982, 524983, 524984, 524985, 524986, 524987, 524988, 551919, 551921, 551922, 551924, 551925, 551926, 55 1933, 551941, 551950, 551951, 551952, 551953, 551955, 551956, 551957, 551958, 551966, 551983, 551984, 551985, 551986, 55198 7, 551989, 551990, 551992, 551993, 551994, 551995, 551996, 551997, 551998, 551999, 552000, 552005, 552006, 552009, 552012, 552013, 552014, 552015, 552017, 552018, 552019, 552020, 552021, 552022, 552023, 552024, 552025, 552026, 552027, 552028, 552 029, 552030, 552031, 552032, 552033, 552034, 552038, 552039, 552041, 552044, 552046, 552047, 552049, 552050, 552051, 552052 ,552053,552054,552055,552056,552057,552058,552059,552060,552061,552062,552063,552064,552065,552068,552069,5 52070, 552071, 552073, 552074, 552075, 552076, 552077, 552078, 552079, 552080, 552081, 552082, 552083, 552084, 552085, 5520 86, 552087, 552088, 552089, 552090, 552091, 552092, 552093, 552094, 552095, 552096, 552097, 552098, 552099, 552100, 552101, 552115, 552117, 552123, 552125, 552127, 552128, 552129, 552132, 552133, 552138, 552139, 552140, 552141, 552143, 552144, 55 2145, 552146, 552147, 552148, 552149, 552150, 552151, 552152, 552158, 552159, 552160, 552163, 552168, 552179, 552187, 55218 8, 552192, 552193, 552195, 552199, 552200, 552201, 552202, 552203, 552204, 552205, 552206, 552207, 552208, 552210, 552211, 552213, 552214, 552222, 552246, 552247, 552248, 552253, 552254, 552255, 552258, 552294, 552301, 552302, 552306, 552307, 552 308, 552309, 552310, 552312, 552314, 552315, 552317, 552318, 552321, 552322, 552323, 552325, 552332, 552337, 552339, 552347 ,552348,552349,552354,552355,552358,552359,552360,552361,552362,552363,552364,552365,552366,552367,552368,5 52369, 552371, 552373, 552374, 552375, 552376, 552377, 552378, 552379, 552403, 552408, 552409, 552411, 552418, 552419, 5524 20, 552424, 552442, 552464, 552465, 552466, 552467, 552472, 552474, 552475, 552477, 552478, 552521, 552522, 552523, 552527, 552528, 552529, 552530, 552534, 552567, 552578, 552579, 552584, 552586, 552587, 552588, 552590, 552789, 552803, 552804, 55 2805, 552808, 552816, 552817, 552818, 552819, 552820, 552821, 552822, 552823, 552824, 552828, 552829, 552830, 552833, 55283 4, 552835, 552842, 552843, 552844, 552846, 552848, 552849, 552850, 552851, 552852, 552853, 552854, 552855, 552856, 552857, 552858, 552859, 552860, 552861, 552863, 552864, 552865, 552872, 552894, 552903, 552904, 552907, 552909, 552910, 552911, 552 913, 552914, 552915, 552916, 552917, 552918, 552922, 552923, 552925, 552927, 552928, 552929, 552930, 552931, 552932, 552933 ,552934,552935,552936,552937,552938,552939,552940,552941,552942,552943,552944,552945,552946,552957,552961,5 52966, 552969, 552971, 552972, 552974, 552976, 552979, 552980, 552981, 552983, 552984, 552988, 552989, 552990, 552991, 5529 95, 552996, 552998, 552999, 553001, 553002, 553003, 553004, 553006, 553008, 553009, 553010, 553011, 553012, 566828, 566829, 566830, 566831, 566832, 577120, 577121, 577122, 577123, 577124, 577125, 577126, 577127, 577128, 577129, 577130, 577131, 57 7132, 577133, 577134, 577135, 577136 and 582666.

[000280] In certain embodiments, the following antisense compounds or oligonucleotides target a region of an HBV nucleic acid and effect at least 70% inhibition of an HBV mRNA, SEQ ID NOs: 12, 17, 18, 20, 21, 22, 24, 25, 26, 27, 28, 29, 39, 40, 43, 1 45, 147, 149, 151, 153, 166, 167, 168, 174, 176, 177, 178, 179, 181, 186, 187, 188, 190, 198, 201, 207, 209, 210, 211, 212, 213, 224, 225, 226, 227, 232, 234, 240, 321, 324, 325, 326, 327, 329, 330, 331, 332, 335, 336, 337, 339, 342, 343, 344, 34 5, 346, 357, 358, 359, 363, 364, 368, 370, 371, 372, 375, 376, 377, 378, 379, 382, 383, 384, 387, 388, 389, 390, 391, 392, 395, 397, 400, 401, 402, 403, 404, 405, 409, 410, 412, 413, 417, 418, 420 421, 422, 423, 424, 425, 426, 427, 430, 435, 436, 438, 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 457, 459, 462, 463, 464, 465, 466, 467, 468, 469, 470, 473, 474, 47 5, 478, 479, 480, 481, 482, 483, 484, 485, 487, 488, 489, 490, 492, 496, 497, 500, 501, 504, 505, 508, 510, 512, 513, 514, 515, 516, 517, 520, 521, 524, 525, 526, 527, 528, 529, 530, 531, 539, 543, 544, 545, 546, 547, 548, 551, 552, 553, 554, 555 ,556,557,558,559,560,561,566,567,569,571,572,573,574,575,576,577,578,579,580,588,589,590,595,596,5 97, 598, 599, 600, 601, 602, 605, 606, 608, 609, 610, 611, 612, 613, 614, 615, 616, 617, 618, 619, 623, 624, 625, 626, 627, 628, 631, 632, 634, 636, 637, 638, 639, 640, 641, 642, 643, 644, 645, 646, 647, 648, 649, 650, 652, 653, 654, 657, 658, 65 9, 660, 661, 662, 663, 665, 666, 667, 669, 671, 672, 673, 674, 675, 676, 677, 678, 679, 680, 682, 684, 686, 687, 688, 689, 690, 691, 692, 693, 694, 695, 696, 698, 699, 700, 701, 702, 703, 704, 705, 706, 707, 708, 709, 710, 711, 712, 713, 714, 715 ,716,717,718,719,720,721,722,723,724,725,726,727,728,729,730,731,733,734,735,736,737,738,740,741,7 42, 745, 754, 755, 756, 757, 768, 777, 778, 779, 780, 781, 782, 784, 790, 791, 792, 793, 794, 795, 796, 797, 798, 799, 800, 801, 802, 803, 804, 805, 806, 807, 808, 809, 810, 811, 812, 814, 815, 817, 818, 819, 820, 821, 822, 823, 824, 825, 826, 82 7,828,833,834,842,843,844,849,852,854,860,892,893,894,895,896,897,898,899,900,1288,1289,1290,1291,1 292, 1293, 1294, 1295, 1296, 1297, 1298, 1299, 1300, 1301, 1302, 1303, 1304, 1305, 1306, 1307, 1308, 1309, 1310, 1311, 1312 ,1313,1314,1315,1316,1317,1320,1322,1323,1324,1325,1326,1327,1329,1330,1331,1332,1333,1334,1335,1336,1 337, 1338, 1340, 1341, 1342, 1343, 1344, 1345, 1346, 1347, 1348, 1349, and 1350, 1367, 1368, 1369, 1370, 1372 and 1376.

[000281] In certain embodiments, the following antisense compounds or oligonucleotides target a region of an HBV nucleic acid and effect at least 80% inhibition of an HBV mRNA, ISIS IDs: 510100, 509931, 510116, 505317, 505319, 505323, 505326, 5053 27, 505330, 505339, 505346, 505347, 505358, 509934, 146786, 524414, 524415, 524416, 524418, 524419, 524425, 524426, 524431, 524432, 524434, 524446, 524447, 524452, 524459, 524460, 524466, 524469, 524475, 524477, 524478, 524479, 524482, 524485, 52 4490, 524491, 524492, 524493, 524494, 524495, 524499, 524502, 524503, 524507, 524510, 524511, 524512, 524520, 524525, 52452 8, 524532, 524533, 524534, 524535, 524536, 524540, 524541, 524547, 524552, 524553, 524556, 524561, 524564, 524565, 524568, 524570, 524571, 524572, 524573, 524578, 524580, 524586, 524590, 524591, 524594, 524595, 524602, 524604, 524606, 524607, 524 610, 524611, 524614, 524616, 524617, 524618, 524619, 524620, 524621, 524633, 524634, 524635, 524636, 524637, 524641, 524643 ,524644,524646,524649,524650,524651,524657,524662,524664,524667,524670,524678,524679,524680,524686,524688,5 24690, 524691, 524692, 524695, 524698, 524699, 524701, 524702, 524704, 524705, 524706, 524707, 524708, 524709, 524713, 5247 15, 524716, 524717, 524718, 524721, 524726, 524727, 524728, 524729, 524730, 524731, 524733, 524734, 524735, 524737, 524739, 524741, 524742, 524743, 524747, 524748, 524749, 524751, 524752, 524754, 524758, 524760, 524762, 524763, 524764, 524767, 52 4768, 524769, 524771, 524773, 524777, 524778, 524779, 524780, 524781, 524783, 524784, 524788, 524789, 524791, 524792, 52479 3, 524794, 524795, 524796, 524797, 524798, 524801, 524803, 524804, 524805, 524806, 524807, 524808, 524809, 524810, 524811, 524813, 524816, 524819, 524822, 524823, 524824, 524827, 524828, 524829, 524833, 524842, 524844, 524880, 524881, 524882, 524 884, 524886, 524887, 524888, 524889, 524890, 524891, 524893, 524907, 524908, 524980, 524986, 524987, 551921, 551924, 551925 ,551953,551956,551957,551984,551986,551987,551989,551990,551993,551994,551995,551996,551997,551998,551999,5 52000, 552005, 552006, 552018, 552019, 552020, 552021, 552022, 552023, 552024, 552025, 552026, 552027, 552028, 552029, 5520 30, 552031, 552032, 552033, 552034, 552039, 552044, 552046, 552050, 552051, 552052, 552053, 552054, 552055, 552056, 552057, 552058, 552059, 552060, 552061, 552062, 552063, 552064, 552065, 552073, 552077, 552078, 552079, 552080, 552082, 552083, 55 2084, 552085, 552086, 552087, 552088, 552089, 552090, 552091, 552092, 552093, 552094, 552095, 552096, 552097, 552098, 55213 8, 552139, 552145, 552146, 552147, 552149, 552192, 552193, 552199, 552200, 552201, 552207, 552246, 552247, 552253, 552301, 552307, 552308, 552310, 552317, 552347, 552348, 552354, 552355, 552360, 552361, 552362, 552363, 552364, 552365, 552366, 552 367, 552371, 552375, 552464, 552465, 552521, 552808, 552816, 552817, 552818, 552819, 552820, 552822, 552824, 552834, 552844 552849 552850 552851 552852 552853 552854 552916 552922 552923 552925 552930 552931 552932 552933 552936 5 52937, 552938, 552939, 552942, 552943, 552944, 552980, 552988, 552989, 552996, 552998, 553002, 553003, 566828, 566829, 5668 30, 566831, 566832, 577120, 577121, 577122, 577123, 577124, 577125, 577126, 577127, 577128, 577130, 577131, 577132, 577133, 577134, 577135, 577136 and 582666.

[000282] In certain embodiments, the following antisense compounds or oligonucleotides target a region of an HBV nucleic acid and effect at least 80% inhibition of an HBV mRNA, SEQ ID NOs: 17, 20, 22, 24, 26, 28, 39, 40, 50, 51, 83, 89, 103, 123, 1 26, 127, 136, 137, 143, 147, 149, 168, 176, 177, 178, 179, 187, 188, 210, 211, 212, 224, 225, 226, 227, 232, 325, 326, 327, 329, 330, 336, 337, 342, 343, 345, 357, 358, 363, 370, 371, 376, 379, 387, 388, 389, 392, 395, 400, 401, 402, 403, 404, 40 5, 409, 412, 413, 417, 420 421, 422, 430, 435, 438, 442, 443, 444, 445, 446, 450, 451, 457, 462, 463, 466, 474, 475, 478, 4 80, 481, 482, 483, 488, 490, 496, 500, 501, 504, 505, 512, 514, 516, 517, 520, 521, 524, 526, 527, 528, 529, 530, 531, 543, 544, 545, 546, 547, 551, 553, 554, 555, 559, 560, 561, 567, 572, 574, 577, 580, 588, 589, 590, 596, 598, 600, 601, 602, 60 5, 608, 609, 611, 612, 614, 615, 616, 617, 618, 619, 623, 625, 626, 627, 628, 631, 636, 637, 638, 639, 640, 641, 643, 644, 645, 646, 648, 650, 652, 653, 654, 658, 659, 660, 662, 663, 665, 669, 671, 673, 674, 675, 678, 679, 680, 682, 684, 688, 689 ,690,691,692,694,695,699,700,702,703,704,705,706,707,708,709,712,714,715,716,717,718,719,720,721,7 22, 723, 725, 728, 731, 734, 735, 736, 740, 741, 745, 756, 791, 792, 793, 795, 797, 798, 799, 800, 801, 802, 804, 805, 806, 807, 819, 820, 892, 898, 899, 1292, 1293, 1295, 1296, 1301, 1302, 1303, 1304, 1305, 1306, 1307, 1308, 1310, 1312, 1316, 13 and 1376.

[000283] In certain embodiments, the following antisense compounds or oligonucleotides target a region of an HBV nucleic acid and effect at least 90% inhibition of an HBV mRNA, ISIS IDs: 524414, 524415, 524432, 524460, 524466, 524469, 524475, 5244 77, 524493, 524512, 524535, 524540, 524552, 524561, 524572, 524617, 524619, 524634, 524641, 524644, 524657, 524667, 524691, 524698, 524699, 524701, 524706, 524707, 524709, 524713, 524715, 524716, 524718, 524721, 524726, 524729, 524730, 524731, 52 4733, 524734, 524735, 524739, 524743, 524754, 524763, 524764, 524767, 524771, 524780, 524781, 524784, 524788, 524789, 52479 1, 524792, 524793, 524794, 524795, 524796, 524797, 524798, 524801, 524803, 524804, 524805, 524806, 524807, 524808, 524809, 524810, 524811, 524822, 524827, 524842, 551986, 551987, 551989, 552005, 552018, 552019, 552020, 552021, 552022, 552023, 552 025, 552046, 552050, 552051, 552052, 552053, 552054, 552055, 552057, 552082, 552083, 552084, 552085, 552086, 552087, 552088 ,552089,552092,552093,552096,552097,552307,552317,552355,552361,552362,552363,552817,552851,552922,552923,5 66828, 566829, 566830, 566831, 566832, 577120, 577121, 577122, 577123, 577124, 577125, 577126, 577127, 577128, 577130, 5771 31, 577132, 577134, 577135, 577136 and 582666.

[000284] In certain embodiments, the following antisense compounds or oligonucleotides target a region of an HBV nucleic acid and effect at least 90% inhibition of an HBV mRNA, SEQ ID NOs: 17, 24, 50, 51, 137, 143, 147, 176, 211, 212, 224, 226, 227 ,325,326,343,371,376,379,403,422,445,450,462,482,527,529,544,551,554,567,577,601,608,609,611,616,6 17, 619, 623, 625, 626, 628, 631, 636, 639, 640, 641, 643, 644, 645, 646, 650, 654, 665, 674, 675, 678, 682, 691, 692, 695, 699, 700, 702, 703, 704, 705, 706, 707, 708, 709, 712, 714, 715, 716, 717, 718, 719, 720, 721, 722, 723, 735, 801, 804, 80 5, 807, 1296, 1302, 1303, 1304, 1312, 1325, 1326, 1332, 1334, 1340, 1345, 1349 and 1376.

[000285] In certain embodiments, the following antisense compounds or oligonucleotides target a region of an HBV nucleic acid and effect at least 95% inhibition of an HBV mRNA, ISIS IDs: 524619, 524634, 524641, 505339, 524698, 524709, 524718, 5247 31, 524734, 524789, 524791, 524792, 524793, 524794, 524795, 524796, 524797, 524798, 524801, 524803, 524804, 524805, 524806, 505346, 146785, 524807, 505347, 524808, 524809, 524810, 524811, 146786, 525101, 525102, 525103, 525107, 525108, 525109, 52 5110, 525111, 525112, 525113, 525114, 525115, 525116, 525117, 525118, 525119, 525120, 552018, 552050, 552019, 552051, 55202 0, 552052, 551987, 552021, 552053, 552005, 552022, 552054, 551989, 552023, 552055, 552084, 552085, 552086, 552087, 552088, 552361, 552317, 566831, 577123, 577124, 566830, 566828, 566829, 577127, 577135, 577132, 577136, 566832 and 577122.

[000286] In certain embodiments, the following antisense compounds or oligonucleotides target a region of an HBV nucleic acid and effect at least 95% inhibition of an HBV mRNA, SEQ ID NOs: 17, 50, 137, 143, 187, 210, 212, 224, 529, 544, 551, 608, 6 19, 628, 641, 645, 700, 702, 703, 704, 705, 706, 707, 708, 709, 712, 715, 716, 717, 718, 719, 720, 721, 722,723, 1014, 1015 ,1016,1020,1021,1022,1023,1024,1025,1026,1027,1028,1029,1030,1031,1032,1033,1236,1302,1312,1334,1340,1 345, 1349.

[000287] Certain embodiments provide methods of treating an HBV-related disease, disorder, or condition in an animal, comprising administering to an animal in need thereof a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide consisting of 10 to 30 linked nucleosides and having a nucleobase sequence comprising at least 10 contiguous nucleobases from any of the amino acid sequences of SEQ ID NOs: 5-310, 321-802, 804-1272, 1288-1350, 1364-1372, 1375, 1376, and 1379.

[000288] Certain embodiments provide methods of treating an HBV-related disease, disorder, or condition in an animal, comprising administering to an animal in need thereof a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide consisting of 10 to 30 linked nucleosides and having a nucleobase sequence comprising at least 10 contiguous nucleobases from any of the amino acid sequences of SEQ ID NOs: 17, 51, 86, 93, 95, 98, 100, 102, 104, 106, 109, 112, 115, 117, 137, 140, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 167, 168, 176, 177- 179, 181, 188, 190- 192, 194, 199 . , 1376, and 1379, wherein at least one nucleoside of the modified oligonucleotide comprises at least one 2'-O-methoxyethyl sugar and/or restricted ethyl (cEt) sugar. In certain embodiments, the modified oligonucleotide is 16 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, or 4 modified sugar nucleosides.

[000289] Certain embodiments provide a method of reducing HBV expression in an animal comprising administering to the animal a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to HBV and having a nucleobase sequence comprising at least 10 contiguous nucleobases from any one of the amino acid sequences of SEQ ID NOs: 5310, 321-802, 804-1272, 1288-1350, 1364-1372, 137 5, 1376, and 1379.

[000290] Certain embodiments provide a method of reducing HBV expression in an animal comprising administering to the animal a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to HBV and having a nucleobase sequence comprising at least 10 contiguous nucleobases from any one of the amino acid sequences of SEQ ID NOs: 17, 51, 86, 93, 95, 98, 100, 102, 104, 106, 109, 1 12, 115, 117, 137, 140, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 167, 168, 176, 177179, 181, 188, 190-192, 194, 1 13 75, and 1376, wherein at least one nucleoside of the modified oligonucleotide comprises at least one 2'-O-methoxyethyl sugar and/or restricted ethyl (cEt) sugar. In certain embodiments, the modified oligonucleotide is 16 nucleosides in length and has a gap segment of 9 or 10 linked nucleosides. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, 4, or 5 modified sugar nucleosides.

[000291] Certain embodiments provide a method for preventing, alleviating or treating an HBV-related disease, disorder or condition in an animal comprising administering to the animal a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to HBV and having a nucleobase sequence comprising at least 10 contiguous nucleobases from any one of the amino acid sequences of SEQ ID NOs: 17, 51, 86, 93, 95, 98, 100, 102, 104, 106, 109, 1 12, 115, 117, 137, 140, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 167, 168, 176, 177- 179, 181, 188, 190- 192, 194, 199, 201, 208, 209, 211, 226, 230-237, 244, 245, 247, 252, 254, 256, 258, 260, 262, 264, 266, 271, 1318-1347, 1364-1372, 1375, and 1376, wherein at least one nucleoside of the modified oligonucleotide comprises at least one 2'-O-methoxyethyl sugar and/or restricted ethyl (cEt) sugar. In certain embodiments, the modified oligonucleotide is 16 nucleosides in length and has a gap segment of 9 or 10 linked nucleosides. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, 4, or 5 modified sugar nucleosides.

[000292] Certain embodiments provide methods of treating an HBV-related disease, disorder, or condition in an animal, comprising administering to an animal in need thereof a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide consisting of 10 to 30 linked nucleosides and having a nucleobase sequence comprising at least 10 contiguous nucleobases of any one of the amino acid sequences of SEQ ID NOs: 5, 15, 16, 33, 39-95, 123-135, 163-175, 180-310, 321-4 06, 413-455, 461-802, or 804-1272.

[000293] Certain embodiments provide methods of treating an HBV-related disease, disorder, or condition in an animal, comprising administering to an animal in need thereof a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide consisting of 10 to 30 linked nucleosides and having a nucleobase sequence comprising at least 10 contiguous nucleobases from any of the amino acid sequences of SEQ ID NOs: 6-14, 17-32, 34-38, 96-122, 136-162, 176-179, 407-412, 456462, 523-538 wherein at least one nucleoside of the modified oligonucleotide comprises at least one 2'-O-methoxyethyl sugar.

[000294] In certain embodiments, the modified oligonucleotide is 14 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-3 or 2 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 16 nucleosides in length and has a gap segment of 9 or 10 linked nucleosides. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, 2-4 or 3 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, 4, or 5 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 17 nucleosides in length and has a gap segment of 9 or 10 linked nucleosides. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, 2-4 or 3-4 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, 4, 5, or 6 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 18 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, 3-5, or 4 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 20 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, or 5 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, 4, 5, 6, 7, or 8 modified sugar nucleosides.

[000295] Certain embodiments provide a method of reducing HBV expression in an animal comprising administering to the animal a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to HBV and having a nucleobase sequence comprising at least 10 contiguous nucleobases from any of the amino acid sequences of SEQ ID NOs: 5310, 321-802, 804-1272, or 1288-1350.

[000296] Certain embodiments provide a method of reducing HBV expression in an animal comprising administering to the animal a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to HBV and having a nucleobase sequence comprising at least 10 contiguous nucleobases from any one of the amino acid sequences of SEQ ID NOs: 5, 15, 16, 33, 39-95, 123-135, 163-175, 180-310, 3 21-406, 413-455, 461802, or 804-1272.

[000297] Certain embodiments provide a method of reducing HBV expression in an animal comprising administering to the animal a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to HBV and having a nucleobase sequence comprising at least 10 contiguous nucleobases from any one of the amino acid sequences of SEQ ID NOs: 614, 17-32, 34-38, 96-122, 136-162, 176-179, 407-4 12, 456-462, 523538 wherein at least one nucleoside of the modified oligonucleotide comprises at least one 2'-O-methoxyethyl sugar.

[000298] In certain embodiments, the modified oligonucleotide is 14 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-3 or 2 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 16 nucleosides in length and has a gap segment of 9 or 10 linked nucleosides. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, 4, or 5 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 16 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, 2-4 or 3 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 17 nucleosides in length and has a gap segment of 9 or 10 linked nucleosides. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, 4, 5, or 6 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 17 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, 2-4 or 3-4 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 18 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, 3-5, or 4 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 20 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, 4, 5, 6, 7, or 8 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 20 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, or 5 modified sugar nucleosides.

[000299] Certain embodiments provide a method for preventing, alleviating or treating an HBV-related disease, disorder or condition in an animal comprising administering to the animal a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to HBV. In certain embodiments, the compound or composition comprises a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to HBV and having a nucleobase sequence comprising at least 10 contiguous nucleobases from any one of the amino acid sequences of SEQ ID NOs: 5310, 321-802, 804-1272, or 1288-1350, wherein at least one nucleoside of the modified oligonucleotide comprises at least one 2'-O-methoxyethyl sugar. In certain embodiments, the modified oligonucleotide has a nucleobase sequence comprising at least 10 contiguous nucleobases from any of the amino acid sequences of SEQ ID NOs: 5-310, 321-802, or 804-1272. In certain embodiments, the modified oligonucleotide has a nucleobase sequence comprising at least 10 contiguous nucleobases from any of the amino acid sequences of SEQ ID NOs: 5, 15, 16, 33, 39-95, 123-135, 163-175, 180-310, 321-406, 413-455, 461-802, or 804-1272. In certain embodiments, the modified oligonucleotide has a nucleobase sequence comprising at least 10 contiguous nucleobases from any of the amino acid sequences of SEQ ID NOs: 614, 17-32, 34-38, 96-122, 136-162, 176-179, 407-412, 456-462, 523538 wherein at least one nucleobase The modified oligonucleotide side comprises at least one 2'-O-methoxyethyl sugar. In certain embodiments, the modified oligonucleotide is 14 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-3 or 2 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 16 nucleosides in length and has a gap segment of 9 or 10 linked nucleosides. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, 4, or 5 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 16 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, 2-4 or 3 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 17 nucleosides in length and has a gap segment of 9 or 10 linked nucleosides. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, 4, 5, or 6 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 17 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, 2-4 or 3-4 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 18 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, 3-5, or 4 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 20 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 2, 3, 4, 5, 6, 7, or 8 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 20 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, or 5 modified sugar nucleosides.

[000300] Examples of HBV-related diseases, disorders, or conditions include, but are not limited to, chronic HBV infection, jaundice, liver cancer, liver inflammation, liver fibrosis, liver cirrhosis, liver failure, diffuse hepatocellular inflammatory disease, hemophagocytic syndrome, serum hepatitis, HBV viremia, and conditions having symptoms that may include any or all of the following: flu-like illness, weakness, aches, headaches, fever, loss of appetite, diarrhea, nausea, and vomiting , pain over the liver area of the body, gray or clay colored stools, itching all over the body, and dark colored urine, when coupled with a positive test for the presence of a hepatitis B virus, a hepatitis B viral antigen, or a positive test for the presence of an antibody specific to a hepatitis B viral antigen.

[000301] Certain embodiments provide a method for reducing HBV mRNA expression in an animal comprising administering to the animal a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to HBV. In certain embodiments, reducing HBV mRNA expression in an animal prevents, alleviates or treats an HBV-related disease, disorder or condition. In certain embodiments, reducing HBV mRNA expression in an animal prevents, alleviates, or treats liver disease. In certain embodiments, HBV mRNA expression is reduced by at least 5%, 10%, 20%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%.

[000302] Certain embodiments provide a method for reducing HBV protein levels in an animal comprising administering to the animal a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to HBV. In certain embodiments, reducing HBV protein levels in an animal prevents, alleviates or treats an HBV-related disease, disorder or condition. In certain embodiments, reducing HBV protein levels in an animal prevents, alleviates, or treats liver disease. In certain embodiments, the HBV protein level is reduced by at least 5%, 10%, 20%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%.

[000303] Certain embodiments provide a method for reducing HBV DNA levels in an animal comprising administering to the animal a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to HBV. In certain embodiments, reducing levels of HBV DNA in an animal prevents, alleviates or treats an HBV-related disease, disorder or condition. In certain embodiments, the mammal can be human, and the hepatitis B virus can be a human hepatitis B virus. More particularly, the human hepatitis B virus can be any of the human geographic genotypes: A (Northwestern Europe, North America, Central America); B (Indonesia, China, Vietnam); C (East Asia, Korea, China, Japan, Polynesia, Vietnam); D (Mediterranean Area, Middle East, India); E (Africa); F (Native Americas, Polynesia); G (United States, France); or H (Central America). In certain embodiments, reducing levels of HBV DNA in an animal prevents, alleviates, or treats liver disease. In certain embodiments, the HBV DNA level is reduced by at least 5%, 10%, 20%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%.

[000304] Certain embodiments provide a method for reducing HBV antigen levels in an animal comprising administering to the animal a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to HBV. In certain embodiments, the antigen is HBsAG or HBeAG. In certain embodiments, reducing HBV antigen levels in an animal prevents, alleviates, or treats an HBV-related disease, disorder, or condition. In certain embodiments, reducing HBV antigen levels in an animal prevents, alleviates, or treats liver disease. In certain embodiments, HBV antigen levels are reduced by at least 5%, 10%, 20%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%.

[000305] Certain embodiments provide a method for reducing HBV and HBV DNA antigen in an animal infected with a hepatitis B virus, comprising administering to the animal a compound or composition described herein. In certain embodiments, the compound or composition comprises a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to HBV. In certain embodiments, the antigen is HBsAG or HBeAG. In certain embodiments, the amount of HBV antigen may be sufficiently reduced to result in seroconversion, defined as absence of serum HBeAg plus presence of HBeAb if monitoring HBeAg as a determinant for seroconversion, or defined as absence of serum HBsAg if monitoring HBsAg as a determinant for seroconversion, as determined by currently available detection limits of commercial ELISA systems.

[000306] Certain embodiments provide a method for treating an animal with an HBV-related disease, disorder or condition comprising: a) identifying said animal with the HBV-related disease, disorder or condition, and b) administering to said animal a therapeutically effective amount of a compound or composition comprising a modified oligonucleotide consisting of 14 to 20 linked nucleosides and having a nucleobase sequence at least 90% complementary to any one of SEQ ID NOs: 1273 ,1274,1275,1276,1277,1278,1279,1280,1281,1282,1283,1284,1285,1287,1352,1353,1354,1359,1360,1361,1362, and 1363, as measured over the entirety of said modified oligonucleotide. In certain embodiments, the therapeutically effective amount of the compound or composition administered to the animal treats or reduces the HBV-related disease, disorder or condition, or a symptom thereof, in the animal. In certain embodiments, the HBV-related disease, disorder or condition is liver disease. In certain embodiments, the related disease, disorder, or condition is chronic HBV infection, jaundice, liver cancer, liver inflammation, liver fibrosis, cirrhosis of the liver, liver failure, diffuse hepatocellular inflammatory disease, hemophagocytic syndrome, serum hepatitis, HBV viremia, or transplant-related liver disease.

[000307] Certain embodiments provide a method for treating an animal with an HBV-related disease, disorder or condition comprising: a) identifying said animal with the HBV-related disease, disorder or condition, and b) administering to said animal a therapeutically effective amount of a compound or composition comprising a modified oligonucleotide consisting of 14 to 20 linked nucleosides and having a nucleobase sequence at least 90% complementary to SEQ ID NO: 1, as measured over the entirety of the said modified oligonucleotide. In certain embodiments, the therapeutically effective amount of the compound or composition administered to the animal treats or reduces the HBV-related disease, disorder or condition, or a symptom thereof, in the animal. In certain embodiments, the HBV-related disease, disorder or condition is liver disease. In certain embodiments, the related disease, disorder, or condition is chronic HBV infection, jaundice, liver cancer, liver inflammation, liver fibrosis, cirrhosis of the liver, liver failure, diffuse hepatocellular inflammatory disease, hemophagocytic syndrome, serum hepatitis, HBV viremia, or transplant-related liver disease.

[000308] In certain embodiments, HBV has the sequence as set forth in GenBank Accession NumbersU95551.1 (incorporated herein as SEQ ID NO: 1) or any variant or fragment thereof. In certain embodiments, HBV has truncated portions of the human sequences set forth in SEQ ID NOs: 1273, 1274, 1275, 1276, 1277, 1278, 1279, 1280, 1281, 1282, 1283, 1284, 1285, 1287, 1352, 1353, 1 354, 1359, 1360, 1361, 1362, and 1363.

[000309] In certain embodiments, the animal is a human being.

[000310] In certain embodiments, the compounds or compositions are designated as a first agent. In certain embodiments, the methods comprise administering a first agent and one or more second agents. In certain embodiments, the methods comprise administering a first agent and one or more second agents. In certain embodiments, the first agent and one or more second agents are co-administered. In certain embodiments the first agent and one or more second agents are co-administered sequentially or concomitantly.

[000311] In certain embodiments, the one or more second agent is also a compound or composition described herein. In certain embodiments, the one or more second agents are other than a compound or composition described herein. Examples of one or more second agents include, but are not limited to, an anti-inflammatory agent, chemotherapeutic agent or anti-infective agent.

[000312] In other related embodiments, the additional therapeutic agent may be an HBV agent, an HCV agent, a chemotherapeutic agent, an antibiotic, an analgesic, a non-steroidal anti-inflammatory agent (NSAID), an antifungal agent, an antiparasitic agent, an anti-nausea agent, an anti-diarrheal agent, or an immunosuppressive agent.

[000313] In certain embodiments, the one or more second agent is an HBV agent. In certain embodiments the HBV agent may include, but is limited to, interferon alfa-2b, interferon alfa-2a, and interferon alfacon-1 (pegylated and non-pegylated), ribavirin; an inhibitor of HBV RNA replication; a second antisense oligomer; a therapeutic HBV vaccine; a prophylactic HBV vaccine; lamivudine (3TC); entecavir (ETV); tenofovir diisoproxil fumarate (TDF); telbivudine (LdT); adefovir; or an HBV antibody therapy (monoclonal or polyclonal).

[000314] In certain embodiments, the one or more second agent is an HCV agent. In certain embodiments the HBV agent may include, but is limited to, interferon alfa-2b, interferon alfa-2a, and interferon alfacon-1 (pegylated and non-pegylated); ribavirin; an HCV RNA replicon inhibitor (e.g. ViroPharma VP50406 series); an HCV antisense agent; a therapeutic HCV vaccine; an HCV protease inhibitor; an HCV helicase inhibitor; or an HCV monoclonal or polyclonal antibody therapy.

[000315] In certain embodiments, the one or more second agents is an anti-inflammatory agent (i.e., an inflammation-reducing therapy). In certain modalities, inflammation-reducing therapy may include, but is limited to, a therapeutic lifestyle change, a steroid, an NSAID, or a DMARD. The steroid may be a corticosteroid. The NSAID can be an aspirin, acetaminophen, ibuprofen, naproxen, the COX inhibitors, indomethacin and the like. The DMARD can be a TNF inhibitor, a purine synthesis inhibitor, calcineurin inhibitor, a pyrimidine synthesis inhibitor, a sulfasalazine, methotrexate and the like.

[000316] In certain embodiments, the one or more second agents is a chemotherapeutic agent (ie, an agent for treating cancer). Chemotherapeutic agents may include, but are not limited to, daunorubicin, daunomycin, dactinomycin, doxorubicin, epirubicin, idarubicin, esorubicin, bleomycin, mafosfamide, ifosfamide, cytosine arabinoside, bis-chloroethylnitrosurea, busulfan, mitomycin C, actinomycin D, mithramycin, prednisone, hydroxypro gesterone, testosterone, tamoxifen, dacarbazine, procarbazine, hexamethylmelamine, pentamethylmelamine, mitoxantrone, amsacrine, chlorambucil, methylcyclohexylnitrosurea, nitrogen mustards, melphalan, cyclophosphamide, 6-mercaptopurine, 6-thioguanine, cytarabine (CA), 5-azacytidine, hydroxyurea, deoxycoformycin, 4-hydroxy peroxycyclophosphoramide, 5-fluorouracil (5-FU), 5-fluordeoxyuridine (5-FUdR), methotrexate (MTX), colchicine, taxol, vincristine, vinblastine, etoposide, trimetrexate, teniposide, cisplatin and gemcitabine diethylstilbestrol (DES).

[000317] In certain embodiments, the one or more second agent is an anti-infective agent. Examples of anti-infective agents include, but are not limited to, antibiotics, antifungal drugs, and antiviral drugs.

[000318] In certain embodiments, administration comprises parenteral administration.

[000319] A certain embodiment provides a method for reducing an amount of HBV mRNA, DNA, protein and/or an amount of HBV antigen in a mammal infected with a hepatitis B virus, the method comprising administering a therapeutically effective amount of a pharmaceutical composition as described above to a mammal in need thereof in order to reduce hepatitis B virus infection and hepatitis B antigen, compared to the amount of HBV mRNA, protein and an amount of HBV antigen BV in the mammal before treatment. In some embodiments, the mammal can be human, and the hepatitis B virus can be a human hepatitis B virus. More particularly, the human hepatitis B virus can be any of the human geographic genotypes: A (Northwest Europe, North America, Central America); B (Indonesia, China, Vietnam); C (East Asia, Korea, China, Japan, Polynesia, Vietnam); D (Mediterranean Area, Middle East, India); E (Africa); F (Native Americas, Polynesia); G (United States, France); or H (Central America).

[000320] In certain embodiments, a method is provided for reducing an amount of HBV mRNA, DNA, protein and/or an amount of HBV antigen in a mammal infected with a hepatitis B virus, the method comprising administering a therapeutically effective amount of a pharmaceutical composition as described above to a mammal in need thereof in order to reduce hepatitis B virus infection and hepatitis B antigen, compared to the amount of HBV mRNA, protein and an amount of HBV antigen HBV in the mammal prior to treatment, wherein the amount of mRNA is reduced by at least 70% compared to the amount prior to administration of the modified antisense oligonucleotide. In certain embodiments, a method is provided for reducing an amount of HBV mRNA, DNA, protein and/or an amount of HBV antigen in a mammal infected with a hepatitis B virus, the method comprising administering a therapeutically effective amount of a pharmaceutical composition as described above to a mammal in need thereof in order to reduce hepatitis B virus infection and hepatitis B antigen, compared to the amount of HBV mRNA, protein and an amount of HBV antigen in the mammal prior to treatment , wherein the amount of mRNA is reduced by at least 75% compared to the amount before administration of the modified antisense oligonucleotide. In certain embodiments, a method is provided for reducing an amount of HBV mRNA, DNA, protein and/or an amount of HBV antigen in a mammal infected with a hepatitis B virus, the method comprising administering a therapeutically effective amount of a pharmaceutical composition as described above to a mammal in need thereof in order to reduce hepatitis B virus infection and hepatitis B antigen, compared to the amount of HBV mRNA, protein and an amount of HBV antigen in the mammal prior to treatment , wherein the amount of mRNA is reduced by at least 80% compared to the amount before administration of the modified antisense oligonucleotide. In certain embodiments, a method is provided for reducing an amount of HBV mRNA, DNA, protein and/or an amount of HBV antigen in a mammal infected with a hepatitis B virus, the method comprising administering a therapeutically effective amount of a pharmaceutical composition as described above to a mammal in need thereof in order to reduce hepatitis B virus infection and hepatitis B antigen, compared to the amount of HBV mRNA, protein and an amount of HBV antigen in the mammal prior to treatment , wherein the amount of mRNA is reduced by at least 85% compared to the amount before administration of the modified antisense oligonucleotide. In certain embodiments, a method is provided for reducing an amount of HBV mRNA, DNA, protein and/or an amount of HBV antigen in a mammal infected with a hepatitis B virus, the method comprising administering a therapeutically effective amount of a pharmaceutical composition as described above to a mammal in need thereof in order to reduce hepatitis B virus infection and hepatitis B antigen, compared to the amount of HBV mRNA, protein and an amount of HBV antigen in the mammal prior to treatment , wherein the amount of mRNA is reduced by at least 90% compared to the amount before administration of the modified antisense oligonucleotide. In certain embodiments, a method is provided for reducing an amount of HBV mRNA, DNA, protein and/or an amount of HBV antigen in a mammal infected with a hepatitis B virus, the method comprising administering a therapeutically effective amount of a pharmaceutical composition as described above to a mammal in need thereof in order to reduce hepatitis B virus infection and hepatitis B antigen, compared to the amount of HBV mRNA, protein and an amount of HBV antigen in the mammal prior to treatment , wherein the amount of mRNA is reduced by at least 95% compared to the amount before administration of the modified antisense oligonucleotide. In related methods, the HBV antigen may be HBsAg or it may be HBeAg, and more particularly, the amount of HBV antigen may be sufficiently reduced to result in seroconversion, defined as absence of serum HBeAg plus presence of HBeAb if monitoring HBeAg as a determinant for seroconversion, or defined as absence of serum HBsAg if monitoring HBsAg as a determinant for seroconversion, as determined by currently available detection limits of systems commercial ELISA.

[000321] A certain embodiment provides a method for promoting seroconversion of a hepatitis B virus in a mammal infected with HBV, the method comprising administering a therapeutically effective amount of a pharmaceutical composition as described above to a mammal infected with hepatitis B; monitoring the presence of HBeAg plus HBeAb in a mammalian serum sample, or monitoring the presence of HBsAg in a mammalian serum sample, such that the absence of HBeAg plus the presence of HBeAb in the serum sample if monitoring HBeAg as a determinant of seroconversion, or or the absence of HBsAg in the serum sample if monitoring HBsAg as a determinant of seroconversion, as determined by current detection limits of commercial ELISA systems, is an indication of seroconversion in the mammal.

[000322] Certain embodiments provide the use of a compound or composition as described herein for preventing, alleviating or treating liver disease, or symptom thereof, in an animal. In certain embodiments, the compound or composition comprises a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to HBV. In certain embodiments, the modified oligonucleotide has a nucleobase sequence comprising at least 10 contiguous nucleobases from any of the amino acid sequences of SEQ ID NOs: 17, 51, 86, 93, 95, 98, 100, 102, 104, 106, 109, 112, 115, 117, 137, 140, 1 43, 145, 147, 149, 151, 153, 155, 157, 159, 161, 167, 168, 176, 177-179, 181, 188, 190-192, 194, 199, 201, 208, 209, 211, 226, 230-237, 244, 245, 247, 252, 254, 256, 258, 260, 262, 264, 266, 271, 1318-1347, 1364 1372, 1375, 1376 and 1379.

[000323] In certain embodiments, compounds or compositions as described herein are effective by virtue of having at least one of an in vitro IC50 of less than 250 nM, less than 200 nM, less than 150 nM, less than 100 nM, less than 90 nM, less than 80 nM, less than 70 nM, less than 65 nM, less than 60 nM, less than 55 nM, less than 50 nM, less than 49 nM, less than 47 nM, less than 46 nM, when delivered to HepG2.2.1 cells. In certain embodiments inhibition is measured with the RTS3370 primer probe set, as described herein.

[000324] In certain embodiments, compounds or compositions as described herein are effective by virtue of having at least one of an in vitro IC50 of less than 250 nM, less than 200 nM, less than 100 nM, less than 90 nM, less than 80 nM, less than 70 nM, less than 60 nM, less than 50 nM, less than 40 nM, less than 35 nM, less than 34 nM, less than 33 nM, less than 32 nM, less than 31 nM, when delivered to HepG2.2.1 cells. In certain embodiments inhibition is measured with the RTS3371 primer probe set, as described herein.

[000325] In certain embodiments, compounds or compositions as described herein are effective by virtue of having at least one of an in vitro IC50 of less than 20 µM, less than 10 µM, less than 9.5 µM, less than 9.0 µM, less than 8.5 µM, less than 8.0 µM, less than 7.5 µM, less than 7.0 µM, less than 6.5 µM M, less than 6.0 μM, less than 5.5 μM, less than 5.0 μM, less than 4.5 μM, less than 4.0 μM, less than 3.5 μM, less than 3.0 μM, less than 2.5 μM, when distributed to HepG2.2.1 cells as described herein.

[000326] In certain embodiments, compounds or compositions as described herein are highly tolerable as demonstrated by having at least one of an increase in an ALT or AST value of no more than 4-fold, 3-fold, or 2-fold over saline-treated animals or an increase in liver, spleen, or kidney weight of no more than 30%, 20%, 15%, 12%, 10%, 5%, or 2%. In certain embodiments, compounds or compositions as described herein are highly tolerable as demonstrated by having no increase in ALT or AST over saline-treated subjects. In certain embodiments, compounds or compositions as described herein are highly tolerable as demonstrated by having no increase in liver, spleen, or kidney weights over saline-treated animals. In certain embodiments, these compounds or compositions include ISIS 146779, ISIS 146786, ISIS 505317, ISIS 505329, ISIS 505332, ISIS 505346, ISIS 505347, ISIS 505358, ISIS 509926, ISIS 509927, ISIS 509932, ISIS In certain embodiments, such compounds or compositions include compounds comprising the nucleobase sequence of any one of SEQ ID NOs: 5-310, 321-802, or 804-1272. In certain embodiments, such compounds or compositions include compounds comprising the nucleobase sequence of any one of SEQ ID NOs: 5, 15, 16, 33, 39-95, 123-135, 163-175, 180-310, 321-406, 413-455, 461-802, or 804-1272. In certain embodiments, such compounds or compositions include compounds comprising the nucleobase sequence of any one of SEQ ID NOs: 6-14, 17-32, 34-38, 96-122, 136-162, 176-179, 407-412, 456-462, 523-538 (updated SEQ ID NOs) wherein at least one oligonucleotide nucleoside modified comprises at least one 2'-O-methoxyethyl sugar. In certain embodiments, the modified oligonucleotide is 14 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-3 or 2 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 16 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, 2-4 or 3 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 17 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, 2-4 or 3-4 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 18 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, 3-5, or 4 modified sugar nucleosides. In certain embodiments, the modified oligonucleotide is 20 nucleosides in length and has a gap segment of 10 nucleosides attached. In certain embodiments, the modified oligonucleotide has a wing segment at the 5' end and 3' end of the gap, each independently having 1-5, or 5 modified sugar nucleosides.

[000327] Certain embodiments provide the use of a compound or composition as described herein in the manufacture of a medicament for treating, alleviating, delaying, or preventing an HBV-related disease, disorder, or condition in an animal.

[000328] Certain embodiments provide the use of a compound or composition as described herein in the manufacture of a medicament for treating, alleviating, delaying or preventing liver disease in an animal.

[000329] Certain embodiments provide a kit for treating, preventing, or preventing an HBV-related disease, disorder, or condition, or a symptom thereof, as described herein, wherein the kit comprises: a) a compound or composition as described herein; and optionally b) an additional agent or therapy as described herein. The kit may further include instructions or a label for using the kit to treat, prevent, or ameliorate the HBV-related disease, condition, or disorder.

Antisense Compounds

[000330] Oligomeric compounds include, but are not limited to, oligonucleotides, oligonucleotides, oligonucleotide analogs, oligonucleotide mimetics, antisense compounds, antisense oligonucleotides, and siRNAs. An oligomeric compound can be "antisense" to a target nucleic acid, meaning that it is capable of undergoing hybridization to a target nucleic acid through hydrogen bonding.

[000331] In certain embodiments, an antisense compound has a nucleobase sequence that, when written in the 5' to 3' direction, comprises the reverse complement of the target segment of a target nucleic acid to which it is directed. In certain of these embodiments, an antisense oligonucleotide has a nucleobase sequence that, when written in the 5' to 3' direction, comprises the reverse complement of the target segment of a target nucleic acid to which it is directed.

[000332] In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 10-30 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 12 to 30 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 12 to 22 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 14 to 30 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 14 to 20 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 15 to 30 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 15 to 20 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 16 to 30 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 16 to 20 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 17 to 30 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 17 to 20 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 18 to 30 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 18 to 21 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 18 to 20 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 20 to 30 subunits in length. In other words, such antisense compounds are 12 to 30 linked subunits, 14 to 30 linked subunits, 14 to 20 subunits, 15 to 30 subunits, 15 to 20 subunits, 16 to 30 subunits, 16 to 20 subunits, 17 to 30 subunits, 17 to 20 subunits, 18 to 30 subunits s, 18 to 20 subunits, 18 to 21 subunits, 20 to 30 subunits, or 12 to 22 linked subunits, respectively. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 14 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 16 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 17 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 18 subunits in length. In certain embodiments, an antisense compound targeted to an HBV nucleic acid is 20 subunits in length. In other embodiments, the antisense compound is 8 to 80, 12 to 50, 13 to 30, 13 to 50, 14 to 30, 14 to 50, 15 to 30, 15 to 50, 16 to 30, 16 to 50, 17 to 30, 17 to 50, 18 to 22, 18 to 24 , 18 to 30, 18 to 50, 19 to 22, 19 to 30, 19 to 50, or 20 to 30 linked subunits. In certain of these embodiments, the antisense compounds are 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66 , 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, or 80 linked subunits in length, or a range defined by any two values above. In some embodiments the antisense compound is an antisense oligonucleotide, and the linked subunits are nucleotides.

[000333] In certain embodiments, antisense oligonucleotides targeted to an HBV nucleic acid can be shortened or truncated. For example, a single subunit can be deleted from the 5' end (5' truncation), or alternatively from the 3' end (3' truncation). A shortened or truncated antisense compound targeted to an HBV nucleic acid can have two subunits deleted from the 5' end, or alternatively can have two subunits deleted from the 3' end, of the antisense compound. Alternatively, the deleted nucleosides can be dispersed throughout the antisense compound, for example, in an antisense compound having a 5'-end deleted nucleoside and a 3'-end deleted nucleoside.

[000334] When a single additional subunit is present in an elongated antisense compound, the additional subunit may be located at the 5' or 3' end of the antisense compound. When two or more additional subunits are present, adjacent subunits may be adjacent to each other, for example in an antisense compound having two subunits added to the 5' end (5' addition), or alternatively to the 3' end (3' addition), of the antisense compound. Alternatively, adjacent subunits can be dispersed throughout the antisense compound, for example, in an antisense compound having a subunit added to the 5' end and a subunit added to the 3' end.

[000335] It is possible to increase or decrease the length of an antisense compound, such as an antisense oligonucleotide, and/or introduce mismatched bases without eliminating the activity. For example, in Woolf et al. (Proc. Natl. Acad. Sci. USA 89:7305-7309, 1992), a series of antisense oligonucleotides 13-25 nucleobases in length were tested for their ability to induce cleavage of a target RNA in an oocyte injection model. Antisense oligonucleotides 25 nucleobases long with 8 or 11 base mismatches near the ends of the antisense oligonucleotides were able to direct specific cleavage of the target mRNA, although to a lesser extent than antisense oligonucleotides that contained no mismatches. Likewise, target-specific cleavage was achieved using 13 nucleobase antisense oligonucleotides, including those with one or three mismatches.

[000336] Gautschi et al. (J. Natl. Cancer Inst. 93:463-471, March 2001), demonstrated the ability of an oligonucleotide having 100% complementarity to the bcl-2 mRNA and having 3 mismatches with the bcl-xL mRNA to reduce the expression of both bcl-2 and bcl-xL, in vitro and in vivo. Furthermore, this oligonucleotide demonstrated potent antitumor activity in vivo.

[000337] Maher and Dolnick (Nuc. Acid. Res. 16:3341-3358,1988) tested a series of 14 nucleobase antisense oligonucleotides in tandem, and a 28 and 42 nucleobase antisense oligonucleotides composed of the sequence of two or three of the tandem antisense oligonucleotides, respectively, for their ability to halt translation of Human DHFR in a rabbit reticulocyte assay. Each of the three 14-nucleotide antisense oligonucleotides alone was able to inhibit translation, albeit to a more modest extent than the 28- or 42-nucleotide antisense oligonucleotides.

Antisense Compound Motifs

[000338] In certain embodiments, antisense compounds targeted to an HBV nucleic acid have chemically modified subunits arranged in a pattern, or motifs, to impart to the antisense compounds properties such as enhanced inhibitory activity, increased binding affinity for a target nucleic acid, or resistance to degradation by nucleases in vivo.

[000339] Antisense chimeric compounds typically contain at least one region modified so as to confer increased resistance to nuclease degradation, increased cellular uptake, increased binding affinity for the target nucleic acid, and/or increased inhibitory activity. A second region of a chimeric antisense compound can optionally serve as a substrate for the cellular endonuclease RNase H, which cleaves the RNA strand of an RNA:DNA duplex.

[000340] Antisense compounds having a gapmer motif are considered chimeric antisense compounds. In a gapmer an inner region having a plurality of nucleotides that support RNaseH cleavage is positioned between outer regions having a plurality of nucleotides that are chemically distinct from the nucleosides of the inner region. In the case of an antisense oligonucleotide having a gapmer motif, the gap segment generally serves as a substrate for cleavage with endonucleases, while the wing segments comprise modified nucleosides. In certain embodiments, regions of a gapmer are differentiated by the types of sugar moieties that comprise each distinct region. The types of sugar moieties that are used to distinguish regions of a gapmer can, in some embodiments, include β-D-ribonucleosides, β-D-deoxyribonucleosides, 2'-modified nucleosides (such 2'-modified nucleosides can include 2'-MOE and 2'-O-CH3, among others), and bicyclic sugar-modified nucleosides (such sugar-modified nucleosides bicyclics may include those that have a restricted ethyl). In certain embodiments, the nucleosides in the wings can include various modified sugar moieties, including, for example, 2'-MOE and bicyclic sugar moieties, such as restricted ethyl or LNA. In certain embodiments, the wings may include various modified sugar moieties. In certain embodiments, the wings can include various combinations of 2'-MOE nucleosides, bicyclic sugar moieties, such as ethyl restricted nucleosides or LNA nucleosides, and 2'-deoxynucleosides.

[000341] Each distinct region may comprise uniform sugar moieties, variants, or alternating sugar moieties. The wing-gap-wing motif is often described as "X-Y-Z", where "X" represents wing length-5', "Y" represents gap length, and "Z" represents wing length-3'. "X" and "Z" can comprise uniform, variant, or alternating sugar moieties. In certain embodiments, "X" and "Y" can include one or more 2'-deoxynucleosides. "Y" can comprise 2'-deoxynucleosides. As used here, a gapmer described as "X-Y-Z" has a configuration such that the gap is immediately adjacent to each of the 5'-wing and the 3'-wing. Thus, no intervening nucleotide exists between the 5'-wing and the gap, or the gap and the 3'-wing. Any of the antisense compounds described here can have a gapmer motif. In certain embodiments, "X" and "Z" are the same; in other modalities they are different. In certain embodiments, "Y" is between 8 and 15 nucleosides. X, Y, or Z can be any one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or more nucleosides.

[000342] In certain embodiments, the gapmers provided herein include, for example, 11-mers having a motif of 1-9-1.

[000343] In certain embodiments, the gapmers provided herein include, for example, 12-mers having a motif of 1-9-2, 2-9-1, or 1-10-1.

[000344] In certain embodiments, the gapmers provided herein include, for example, 13-mers having a motif of 1-9-3, 2-9-2, 3-91, 1-10-2, or 2-10-1.

[000345] In certain embodiments, the gapmers provided herein include, for example, 14-mers having a motif of 1-9-4, 2-9-3, 3-92, 4-9-1, 1-10-3, 2-10-2, or 3-10-1.

[000346] In certain embodiments, the gapmers provided herein include, for example, 15-mers having a motif of 1-9-5, 2-9-4, 3-93, 4-9-2, 5-9-1, 1-10-4, 2-10-3, 3-10-2, or 4-10-1.

[000347] In certain embodiments, the gapmers provided herein include, for example, 16-mers having a motif of 4-8-4, 2-9-5, 3-94, 4-9-3, 5-9-2, 1-10-5, 2-10-4, 3-10-3, 4-10-2, 3-8-5, or 5-10-1.

[000348] In certain embodiments, the gapmers provided herein include, for example, 17-mers having a motif of 3-9-5, 3-10-4, 49-4, 5-9-3, 2-10-5, 3-10-4, 4-10-3, 5-10-2, 2-9-6, 5-8-4, 5-7-5, 6-7-4, or 6-9-2.

[000349] In certain embodiments, the gapmers provided herein include, for example, 18-mers having a motif of 4-9-5, 5-9-4, 310-5, 4-10-4, or 5-10-3.

[000350] In certain embodiments, the gapmers provided herein include, for example, 19-mers having a motif of 5-9-5, 4-10-5, or 5-10-4.

[000351] In certain embodiments, the gapmers provided herein include, for example, 20-mers having a motif of 5-10-5, 2-10-8, 810-2, 3-10-7, 7-10-3, 4-10-6, or 6-10-4.

[000352] In certain embodiments, the antisense compound has a "wingmer" motif, having a wing-gap or gap-wing configuration, i.e. an X-Y or Y-Z configuration as described above for the gapmer configuration. Thus, wingmer configurations provided here include, but are not limited to, for example, 5-10, 8-4, 4-12, 12-4, 3-14, 16-2, 18-1, 10-3, 2-10, 1-10, 8-2, 2-13, 5-13, 5-8, or 6-8.

[000353] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 2-10-2 gapmer motif.

[000354] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 3-10-3 gapmer motif.

[000355] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 4-10-4 gapmer motif.

[000356] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 5-10-5 gapmer motif.

[000357] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 3-10-4 gapmer motif.

[000358] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 2-10-4 gapmer motif.

[000359] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 2-10-8 gapmer motif.

[000360] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has an 8-10-2 gapmer motif.

[000361] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 3-10-7 gapmer motif.

[000362] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 7-10-3 gapmer motif.

[000363] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 4-10-6 gapmer motif.

[000364] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 6-10-4 gapmer motif.

[000365] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 2-9-6 gapmer motif.

[000366] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 6-9-2 gapmer motif.

[000367] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 4-9-4 gapmer motif.

[000368] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 5-9-3 gapmer motif.

[000369] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 3-9-5 gapmer motif.

[000370] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 5-9-2 gapmer motif.

[000371] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 2-9-5 gapmer motif.

[000372] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 4-9-3 gapmer motif.

[000373] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a 3-9-4 gapmer motif.

[000374] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a gap-enlarged motif.

[000375] In certain embodiments, the antisense compound targeted to an HBV nucleic acid has a gapmer motif wherein the gap consists of 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 linked nucleosides.

[000376] In certain embodiments, antisense compounds targeting an HBV nucleic acid have any of the following sugar motifs: k-d(10)-k e-d(10)-k k-d(10)-e k-k-d(10)-k-k k-k-d(10)-e-e e-e-d(10)-k-k k-k-k-d(10)-k-k-k e- e-e-d(10)-k-k-k k-k-k-d(10)-e-e-e k-k-k-d(10)-k-k-k e-k-k-d(10)-k-k-e e-e-k-d(10)-k-k-e e-d-k-d(10)-k-k-e e-k-d(10)-k-e-k-e k-d(10) )-k-e-k-e-e e-e-k-d(10)-k-e-k-e e-d-d-k-d(9)-k-k-e e-e-e-e-d(9)-k-k-e e-e-e-e-e-d(10)-e-e-e-e-e k-d-k-d-k-d(9)-e-e e-e-k-k-d(9)-e -k-e-e k-d-k-d-k-d(10)-e-e-e-e-e k-e-k-d(10)-k-e-k e-e-e-k-k-d(8)-e-e-e-e e-e-e-k-k-d(7)-k-k-e-e-e e-e-e-k-d(9)-k-e-e-e e-e-e-k-k -d(7)-k-k-e-e-e e-e-e-e-k-k-d(7)-e-e-e-e e-k-e-k-d(9)-e-e-e-e e-k-e-k-d-k-d(7)-e-e-e-e e-e-e-k-k-d(7)-k-k-e-e-e k-d-k-d-k-d(8)-e -e-e-e-e where, k is an ethyl restricted nucleoside, e is a 2'-MOE substituted nucleoside, and d is a 2'-deoxynucleoside.

[000377] In certain embodiments, the antisense oligonucleotide has a sugar motif described by Formula A as follows: (J)m-(B)n- (J)p-(B)r-(A)t-(D)g-(A)v-(B)w-(J)x-(B)y-(J)z wherein: each A is independently a 2'-substituted nucleoside; each B is independently a bicyclic nucleoside; each J is independently either a 2'-substituted nucleoside or a 2'-deoxynucleoside; each D is a 2'-deoxynucleoside; m is 0-4; n is 0-2; p is 0-2; r is 0-2; t is 0-2; v is 0-2; w is 0-4; x is 0-2; y is 0-2; z is 0-4; g is 6-14; provided that: at least one of m, n, and r is different from 0; at least one of w and y is different from 0; the sum of m, n, p, r, and t is from 2 to 5; and the sum of v, w, x, y, and z is from 2 to 5.

Target Nucleic Acids, Target Regions and Nucleotide Sequences

[000378] Nucleotide sequences encoding HBV include, without limitation, the following: GENBANK Accession U95551.1 (incorporated herein as SEQ ID NO: 1).

[000379] It is understood that the sequence set forth in each SEQ ID NO in the Examples contained herein is independent of any modification to a sugar moiety, an internucleoside linkage, or a nucleobase. As such, antisense compounds defined by SEQ ID NO may independently comprise one or more modifications to a sugar moiety, an internucleoside linkage, or a nucleobase. Antisense compounds described by Isis Number (Isis No.) indicate a combination of motif and nucleobase sequence.

[000380] In certain embodiments, a target region is a structurally defined region of the target nucleic acid. For example, a target region can encompass a 3' UTR, a 5' UTR, an exon, an intron, an exon/intron junction, a coding region, a translation initiation region, translation termination region, or other defined nucleic acid region. Structurally defined regions for HBV can be obtained by accession numbers from sequence databases, such as NCBI, and such information is incorporated herein by reference. In certain embodiments, a target region can span the sequence from a 5' target site of one target segment within the target region to a 3' target site of another target segment within the same target region.

[000381] Targeting includes determining at least one target segment to which an antisense compound hybridizes, such that the desired effect occurs. In certain embodiments, the desired effect is a decrease in mRNA target nucleic acid levels. In certain embodiments, the desired effect is a reduction in levels of the protein encoded by the target nucleic acid or a phenotypic change associated with the target nucleic acid.

[000382] A target region can contain one or more target segments. Multiple target segments within a target region can be superimposed. Alternatively, they may not overlap. In certain embodiments, target segments within a target region are separated by no more than about 300 nucleotides. In certain embodiments, target segments within a target region are separated by a number of nucleotides that is, is about, is not more than, is not more than about, 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 nucleotides in the target nucleic acid, or is a range defined by any of the preceding two values. In certain embodiments, target segments within a target region are separated by no more than, or no more than about, 5 nucleotides in the target nucleic acid. In certain embodiments, the target segments are contiguous. Contemplated are target regions defined by a band having a nucleic acid starting point that is any one of the 5' target sites or 3' target sites indicated herein.

[000383] Suitable target segments can be found within a 5' UTR, a coding region, a 3' UTR, an intron, an exon, or an exon/intron junction. Target segments containing a start codon or a stop codon are also suitable target segments. A suitable target segment specifically excludes a certain structurally defined region as the start codon or stop codon.

[000384] Determining suitable target segments may include a comparison of the sequence of a target nucleic acid with other sequences throughout the genome. For example, the BLAST algorithm can be used to identify regions of similarity between different nucleic acids. This comparison may preclude the selection of antisense compound sequences that can hybridize in a non-specific manner to different sequences of a selected target nucleic acid (i.e., non-target sequences or off-target sequences).

[000385] There may be variations in the activity (eg, as defined by percent reduction in target nucleic acid levels) of antisense compounds within an active target region. In certain embodiments, reduction in HBV mRNA levels is indicative of inhibition of HBV expression. Reductions in levels of an HBV protein are also indicative of inhibition of target mRNA expression. Furthermore, the phenotypic changes are indicative of inhibition of HBV expression. In certain embodiments, reduction of fatigue, reduction of flu-like symptoms, increased appetite, reduction of nausea, reduction of joint pain, reduction of jaundice, reduction of pain in the abdomen, reduction of weakness, reduction of weight loss, reduction of breast swelling in men, reduction of rash on the palms of the hands, reduction of difficulty with blood clotting, reduction of cirrhosis, reduction of spider-like blood vessels in the skin, increased absorption of vitamins A and D, reduction of tumor growth, reduction of tumor volume, reduction of pain headache, reduction of fever, reduction of diarrhea, reduction of pain over the liver area of the body, reduction of clay or gray colored stools, reduction of itching, reduction of dark colored urine, and reduction of nausea and vomiting may be indicative of inhibition of HBV expression. In certain embodiments, amelioration of symptoms associated with HBV-related conditions, diseases and disorders can be indicative of inhibition of HBV expression. In certain embodiments, reduction in cirrhosis is indicative of inhibition of HBV expression. In certain embodiments, the reduction of liver cancer markers may be indicative of inhibition of HBV expression.

Hybridization

[000386] In some embodiments, hybridization occurs between an antisense compound described herein and an HBV nucleic acid. The most common mechanism of hybridization involves hydrogen bonding (eg, Watson-Crick, Hoogsteen, or reversed Hoogsteen hydrogen bonding) between the complementary nucleobases of nucleic acid molecules.

[000387] Hybridization can occur under varying conditions. Stringent conditions are sequence-dependent and are determined by the nature and composition of the nucleic acid molecules to be hybridized.

[000388] Methods for determining whether a sequence is specifically hybridizable to a target nucleic acid are well known in the art. In certain embodiments, the antisense compounds provided herein are specifically hybridized to an HBV nucleic acid.

Complementarity

[000389] An antisense compound and a target nucleic acid are complementary to each other when a sufficient number of nucleobases of the antisense compound can hydrogen bond with the corresponding nucleobases of the target nucleic acid, such that the desired effect occurs (for example, antisense inhibition of a target nucleic acid, such as an HBV nucleic acid).

[000390] Non-complementary nucleobases between an antisense compound and an HBV nucleic acid can be tolerated as long as the antisense compound remains capable of specifically hybridizing to a target nucleic acid. Furthermore, an antisense compound can hybridize over one or more segments of HBV nucleic acid such that adjacent or interfering segments are not involved in the hybridization event (e.g., a loop structure, hairpin structure, or mismatch).

[000391] In certain embodiments, the antisense compounds provided herein, or a portion of information thereof, are, or are at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to an HBV nucleic acid, a target region, target segment, or a specific portion thereof. The percentage complementarity of an antisense compound with a target nucleic acid can be determined using routine methods.

[000392] For example, an antisense compound in which 18 out of 20 nucleobases of the antisense compound are complementary to a target region, and therefore specifically hybridize, would represent 90 percent complementarity. In this example, the remaining non-complementary nucleobases can be clustered or interspersed with complementary nucleobases and need not be contiguous with each other or the complementary nucleobases. As such, an antisense compound that is 18 nucleobases in length having four non-complementary nucleobases that are flanked by two regions of complete complementarity with the target nucleic acid would have 77.8% total complementarity with the target nucleic acid and would therefore fall within the scope of the present invention. The percentage complementarity of an antisense compound with a region of a target nucleic acid can be routinely determined using BLAST programs (basic local alignment search tools) and PowerBLAST programs known in the art (Altschul et al., J. Mol. Biol., 1990, 215, 403-410; Zhang and Madden, Genome Res., 1997, 7, 649-656). percent homology, sequence identity, or complementarity can be determined, for example, by the Gap program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, Madison Wis.), using default settings, which utilize the algorithm of Smith and Waterman (Adv. Appl. Math., 1981, 2, 482-489).

[000393] In certain embodiments, the antisense compounds provided herein, or specific portions thereof, are completely complementary (i.e., 100% complementary) to a target nucleic acid, or specified portion thereof. For example, an antisense compound can be completely complementary to an HBV nucleic acid or target region, or target segment or target sequence thereof. As used herein, "fully complementary" means each nucleobase of an antisense compound that is capable of precise base pairing with the corresponding nucleobases of a target nucleic acid. For example, a 20 nucleobase antisense compound is completely complementary to a target sequence that is 400 nucleobases in length, provided there is a corresponding 20 nucleobase portion of the target nucleic acid that is completely complementary to the antisense compound. Completely complementary can also be used in reference to a specific portion of the first and/or second nucleic acid. For example, a 20 nucleobase portion of a 30 nucleobase antisense compound can be "completely complementary" to a target sequence that is 400 nucleobases in length. The 20 nucleobase portion of the 30 nucleobase oligonucleotide is completely complementary to the target sequence if the target sequence has a corresponding 20 nucleobase portion, where each nucleobase is complementary to the 20 nucleobase portion of the antisense compound. At the same time, the full 30 nucleobase antisense compound may or may not be perfectly complementary to the target sequence, depending on whether the remaining 10 nucleobases of the antisense compound are also complementary to the target sequence.

[000394] The location of a non-complementary nucleobase can be at the 5' end or 3' end of the antisense compound. Alternatively, the non-complementary nucleobase or nucleobases can be at an internal position of the antisense compound. When two or more non-complementary nucleobases are present, they can be contiguous (i.e. linked) or non-contiguous. In one embodiment, a non-complementary nucleobase is located on the wing segment of a gapmer antisense oligonucleotide.

[000395] In certain embodiments, antisense compounds that are, or are up to 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleobases in length comprise no more than 4, no more than 3, more than 2, or no more than 1 nucleobase(s) not complementary to a target nucleic acid, such as a HBV nucleic acid, or a specific portion thereof.

[000396] In certain embodiments, antisense compounds that are or are up to 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases in length comprise not more than 6, not more than 5, not more than 4, not more than 3, not more than 2, or more than 1 nucleobase(s) not complementary to a target nucleic acid, such as an HBV nucleic acid, or a specific portion thereof.

[000397] Antisense compounds provided also include those that are complementary to a portion of a target nucleic acid. As used herein, "portion" refers to a defined number of adjacent (i.e., linked) nucleobases within a target nucleic acid region or segment. A "portion" can also refer to a defined number of contiguous nucleobases of an antisense compound. In certain embodiments, antisense compounds are complementary to at least a nucleobase portion of a target segment. In certain embodiments, the antisense compounds are complementary to at least a nucleobase portion of a target segment. In certain embodiments, the antisense compounds are complementary to at least a 10-nucleotide portion of the target segment. In certain embodiments, the antisense compounds are complementary to at least an 11-nucleobase portion of the target segment. In certain embodiments, the antisense compounds are complementary to at least a 12-nucleotide portion of the target segment. In certain embodiments, the antisense compounds are complementary to at least a 13-nucleotide portion of the target segment. In certain embodiments, the antisense compounds are complementary to at least a 14-nucleotide portion of the target segment. In certain embodiments, the antisense compounds are complementary to at least a 15-nucleotide portion of the target segment. Also contemplated are antisense compounds that are complementary to at least a 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more nucleobase portion of a target segment, or a range defined by any two of these values.

Identity

[000398] Antisense compounds provided herein may also have a percent identity to a particular nucleotide sequence, SEQ ID NO, or compound represented by a specific ISIS number, or portion thereof. As used herein, an antisense compound is identical to the sequence disclosed herein if it has the same nucleobase pairing capability. For example, an RNA that contains uracil in place of thymidine in a disclosed DNA sequence could be considered identical to the DNA sequence since both uracil and thymidine pair with adenine. Shortened and lengthened versions of the antisense compounds described herein as well as compounds having non-identical bases relative to the antisense compounds provided herein are also contemplated. Non-identical bases can be adjacent to each other or dispersed throughout the antisense compound. The percentage of identities of an antisense compound is calculated according to the number of bases that have identical base pairing with respect to the sequence to which it is being compared.

[000399] In certain embodiments, the antisense compounds, or portions thereof, are at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to one or more of the antisense compounds or SEQ ID NOs, or a portion thereof, disclosed herein.

[000400] In certain embodiments, a portion of the antisense compound is compared to an equal length portion of the target nucleic acid. In certain embodiments, an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleobase portion is compared to an equal length portion of the target nucleic acid.

[000401] In certain embodiments, a portion of the antisense oligonucleotide is compared to an equal-length portion of the target nucleic acid. In certain embodiments, an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleobase portion is compared to an equal length portion of the target nucleic acid.

Modifications

[000402] A nucleoside is a base-sugar combination. The nucleobase (also known as the base) portion of the nucleoside is usually a heterocyclic base portion. Nucleotides are nucleosides that further include a phosphate group covalently attached to the sugar portion of the nucleoside. For nucleosides that include a pentofuranosyl sugar, the phosphate group can be attached to the 2', 3', or 5' hydroxyl portion of the sugar. Oligonucleotides are formed by covalently linking adjacent nucleosides together to form a linear polymeric oligonucleotide. Within the structure of the oligonucleotide, the phosphate groups are commonly referred to as forming the internucleoside linkages of the oligonucleotide.

[000403] Modifications to antisense compounds include substitutions or changes to internucleoside linkages, sugar moieties, or nucleobases. Modified antisense compounds are often preferred over native forms because of desirable properties, such as, for example, enhanced cellular uptake, enhanced affinity for the target nucleic acid, increased stability in the presence of nucleases, or increased inhibitory activity.

[000404] Chemically modified nucleosides can also be employed to increase the binding affinity of a shortened or truncated antisense oligonucleotide for its target nucleic acid. Therefore, comparable results can often be obtained with shorter antisense compounds that have such chemically modified nucleosides.

Modified Internucleoside Linkages

[000405] The naturally occurring internucleoside linkage of RNA and DNA is a 3' to 5' phosphodiester. Antisense compounds having one or more modified internucleoside linkages, i.e., non-naturally occurring internucleoside linkages, are often chosen over antisense compounds having naturally occurring internucleoside linkages because of desirable properties, such as, for example, enhanced cellular uptake, enhanced affinity for target nucleic acids, and increased stability in the presence of nucleases.

[000406] Oligonucleotides having modified internucleoside linkages include internucleoside linkages that retain a phosphorus atom, as well as internucleoside linkages that lack a phosphorus atom. Representative phosphors containing internucleoside linkages include, but are not limited to, phosphodiesters, phosphotriesters, methylphosphonates, phosphoramidate, and phosphorothioates. Methods of preparing phosphorus-containing and non-phosphorus-containing linkages are well known.

[000407] In certain embodiments, antisense compounds targeted to an HBV nucleic acid comprise one or more modified internucleoside linkages. In certain embodiments, the modified internucleoside linkages are phosphorothioate linkages. In certain embodiments, each internucleoside linkage of an antisense compound is a phosphorothioate internucleoside linkage.

Modified Sugar Servings

[000408] The antisense compounds provided herein may optionally contain one or more nucleosides in which the sugar group has been modified. Such sugar-modified nucleosides can confer enhanced nuclease stability, increased binding affinity, or any other beneficial biological property to antisense compounds. In certain embodiments, the nucleosides comprise a chemically modified ribofuranose ring moiety. Examples of chemically modified ribofuranose rings include, without limitation, the addition of substituent groups (including 5' and 2' substituent groups), linkage of non-geminal ring atoms to form bicyclic nucleic acids (BNA); replacement of the ribosyl ring oxygen atom with S, N(R), or C(R1)(R)2 (R = H, C1-C12 alkyl or a protecting group); and combinations thereof. Examples of chemically modified sugars include, 2'-F-5'-methyl substituted nucleoside (see, PCT International Application WO 2008/101157, published 8/21/08 for other disclosed 5', 2'-bis substituted nucleosides), substitution of the ribosyl ring oxygen atom with S with additional substitution at the 2'-position (see, Published US Patent Application US2005/0130923, published June 16, 2005), or, alternatively, 5'-substitution of a BNA (see, PCT International Application WO 2007/134181, published 11/22/07, where LNA is substituted with, for example, a 5'-methyl group or a 5'-vinyl group).

[000409] Examples of nucleosides having modified sugar moieties include, without limitation, nucleosides comprising 5'-vinyl, 5'-methyl (R or S), 4'-S, 2'-F, 2'-OCH3, and 2'-O(CH2)2OCH3 substituent groups. The substituent in position 2 'can also be selected from alila, amino, pounded, uncle, o-alila, o-c1-c10 alkyl, ocf3, o (CH2) 2sch3, o (CH2) 2-o-n (rm), and o-ch2-c (= o) -n (rm) (rn), where each NM and RN is independently, h or c1-c10 or not replaced.

[000410] As used herein, "bicyclic nucleosides" refer to modified nucleosides comprising a bicyclic sugar moiety. Examples of bicyclic nucleosides include, without limitation, nucleosides comprising a bridge between the 4' and 2' ribosyl ring atoms. In certain embodiments, antisense compounds provided herein include one or more bicyclic nucleosides wherein the bridge comprises a 4' to 2' bicyclic nucleoside. Examples of such 4' to 2' bicyclic nucleosides include, but are not limited to, one of the formulas: 4'-(CH2)-O-2' (LNA); 4'-(CH2)-S-2'; 4'-(CH2)2-O-2' (ENA); 4'-CH(CH3)-O-2' (cEt) and 4'-CH(CH2OCH3)-O-2', and analogues thereof (see, US Patent 7,399,845, Issued July 15, 2008); 4'-C(CH3)(CH3)-O-2', and analogues thereof (see, Published International Application PCT WO2009/006478, published January 8, 2009); 4'-CH2-N(OCH3)-2', and analogues thereof (see, Published International Application PCT WO2008/150729, published December 11, 2008); 4'-CH2-O-N(CH3)-2' (vide, published US Patent Application US2004/0171570, published September 2, 2004); 4'-CH2-N(R)-O-2', where R is H, C1-C12 alkyl, or a protecting group (see, US Patent 7,427,672, Issued September 23, 2008); 4'-CH2-C(H)(CH3)-2' (see, Chattopadhyaya, et al., J. Org. Chem., 2009, 74, 118-134 ); and 4'-CH2-C(=CH2)-2', and analogues thereof (see, Published International Application PCT WO 2008/154401, published December 8, 2008). See also, for example: Singh et al., Chem. Commun., 1998, 4, 455-456; Koshkin et al., Tetrahedron, 1998, 54, 3607-3630; Wahlestedt et al., Proc. Natl. academic Sci. U.S.A., 2000, 97, 5633-5638; Kumar et al., Bioorg. Med. chem. Lett., 1998, 8, 2219-2222; Singh et al., J. Org. Chem., 1998, 63, 10035-10039; Srivastava et al., J. Am. chem. Soc., 129(26) 8362-8379 (Jul. 4, 2007); Elayadi et al., Curr. Opinion Invens. Drugs, 2001, 2, 558561; Braasch et al., Chem. Biol., 2001, 8, 1-7; Orum et al., Curr. Opinion Mol. Ther., 2001, 3, 239-243; US Patent 6,670,461, 7,053,207, 6,268,490, 6,770,748, 6,794,499, 7,034,133, 6,525,191, 7,399,845; Published International Application PCTs WO 2004/106356, WO 94/14226, WO 2005/021570, and WO 2007/134181; US Patent Publications US2004/0171570, US2007/0287831, and US2008/0039618; and US Patents 12/129,154, 60/989,574, 61/026,995, 61/026,998, 61/056,564, 61/086,231, 61/097,787, and 61/099,844; and PCT International Application Nos. PCT/US2008/064591, PCT/US2008/066154, and PCT/US2008/068922. Each of the foregoing bicyclic nucleosides can be prepared having one or more sugar stereochemical configurations including, for example, α-L-ribofuranose and β-D-ribofuranose (see, PCT International Application PCT/DK98/00393, published March 25, 1999 as WO 99/14226).

[000411] In certain embodiments, bicyclic sugar moieties of BNA nucleosides include, but are not limited to, compounds having at least one bridge between the 4' and 2' position of the pentofuranosyl sugar moiety wherein such bridges independently comprise 1 or 2 to 4 independently linked groups selected from -[C(Ra)(Rb)]n-, -C(Ra)=C(Rb)-, -C(Ra)=N- , -C(=NRa)-, -C(=O)-, -C(=S)-, -O-, -Si(Ra)2-, -S(=O)x-, and -N(Ra)-; where: x is 0, 1, or 2; n is 1, 2, 3, or 4; each Ra and Rb is independently H, a protecting group, hydroxyl, C1-C12 alkyl, substituted C1-C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkylkynyl, C5-C20 aryl, substituted C5-C20 aryl, heterocycle radical, substituted heterocycle radical, heteroaryl, substituted heteroaryl , C5C7 acyclic radical, C5-C7 substituted acyclic radical, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)2-J1), or sulfoxyl (S(=O)-J1); and each of J1 and J2 is, independently, H, C1-C12 alkyl, substituted C1C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkynyl, C5-C20 aryl, substituted C5-C20 aryl, acyl (C(=O)-H), substituted acyl, a heterocyclic radical, a substituted heterocyclic radical, C1-C12 aminoalkyl, substituted C1-C12 aminoalkyl, or a protecting group.

[000412] In certain embodiments, the bridge of a bicyclic sugar moiety is, -[C(Ra)(Rb)]n-, -[C(Ra)(Rb)]n-O-, -C(RaRb)-N(R)-O-, or, —C(RaRb)-O-N(R)-. In certain embodiments, the bridge is 4'-CH2-2', 4'-(CH2)2-2', 4'-(CH2)3-2', 4'-CH2-O-2', 4'-(CH2)2-O-2', 4'-CH2-O-N(R)-2', and 4'-CH2-N(R)-O-2'-, where each R is independently H, a protecting group, or C1-C12 alkyl.

[000413] In certain embodiments, bicyclic nucleosides are further defined by isomeric configuration. For example, the nucleoside comprising a 4'-2' methylene-oxy bridge can be in either the α-L n configuration or the β-D configuration. Previously, α-L-methylene-oxy (4'-CH2-O-2') BNA was incorporated into antisense oligonucleotides that showed antisense activity ( Frieden et al., Nucleic Acids Research, 2003, 21, 6365-6372 ).

[000414] In certain embodiments, bicyclic nucleosides include, but are not limited to, (A) α-Methylene-oxy BNA (4'-CH2-O-2'), (B) β-D-Methylene-Oxy BNA (4'-CH2-O-2'), (C) Ethylene-oxy BNA (4'-(CH2)2-O-2'), (D) Aminooxy BNA (4'-CH2-ON(R)-2'), (E) Oxyamino BNA (4'-CH2-N(R)-O-2'), (F) Methyl(methylene-oxy) (4'-CH(CH3)-O-2') BNA), (G) Methylene-thio BNA (4'-CH2-S-2'), (H) Methylene-amino BNA (4'-CH2-N(R)-2'), (I) BNA of carbocyclic methyl (4'-CH2-CH(CH3)-2'), and (J) carbocyclic propylene (4'-(CH2)3-2') BNA as depicted below. where Bx is the base moiety and R is independently H, a protecting group, or C1-C12 alkyl.

[000415] In certain embodiments, the bicyclic nucleoside having Formula I: wherein: Bx is a heterocyclic base moiety; -Qa-Qb-Qc- is -CH2-N(Rc)-CH2-, -C(=O)-N(Rc)-CH2-, -CH2-O-N(Rc)-, -CH2-N(Rc)-O-, or -N(Rc)-O-CH2; Rc is C1-C12 alkyl or an amino protecting group; and Ta and Tb are each, independently, H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety, or a covalent bond to a support medium.

[000416] In certain embodiments, the bicyclic nucleoside having Formula II: wherein: Bx is a heterocyclic base moiety; Ta and Tb are each independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety, or a covalent bond to a support means; Za is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, substituted C1-C6 alkyl, substituted C2-C6 alkenyl, substituted C2-C6 alkynyl, acyl, substituted acyl, substituted amide, thiol, or substituted thio.

[000417] In one embodiment, each of the substituted groups is independently mono- or polysubstituted with substituent groups independently selected from halogen, oxo, hydroxyl, OJc, NJcJd, SJc, N3, OC(=X)Jc, and NJeC(=X)NJcJd, wherein each Jc, Jd, and Je is independently H, C1-C6 alkyl, or C1-C6 alkyl replaced it and X is O or NJc.

[000418] In certain embodiments, the bicyclic nucleoside having Formula III: wherein: Bx is a heterocyclic base moiety; Ta and Tb are each independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety, or a covalent bond to a support means; Zb is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, substituted C1-C6 alkyl, substituted C2-C6 alkenyl, substituted C2-C6 alkynyl or substituted acyl (C(=O)-).

[000419] In certain embodiments, the bicyclic nucleoside having Formula IV: wherein: Bx is a heterocyclic base moiety; Ta and Tb are each independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety, or a covalent bond to a support means; Rd is C1-C6 alkyl, substituted C1-C6 alkyl, C2-C6 alkenyl, substituted C2-C6 alkenyl, C2-C6 alkynyl, or substituted C2-C6 alkynyl; each qa, qb, qc, and qd is, independently, H, halogen, C1C6 alkyl, substituted C1-C6 alkyl, C2-C6 alkenyl, substituted C2-C6 alkenyl, C2-C6 alkynyl, or substituted C2-C6 alkyl, alkoxy, substituted C1-C6 alkoxy, acyl, substituted acyl, aminoalkyl, or substituted C1-C6 aminoalkyl ;

[000420] In certain embodiments, the bicyclic nucleoside Formula V: wherein: Bx is a heterocyclic base moiety; Ta and Tb are each independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety, or a covalent bond to a support means; qa, qb, qe and qf are each independently hydrogen, halogen, C1-C12 alkyl, substituted C1-C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkyl, C1-C12 alkoxy, C1-C12 substituted alkoxy, OJj, SJj, SOJj, SO2 Jj, NJjJk, N3, CN, C(=O)OJj, C(=O)NJjJk, C(=O)Jj, OC(=O)NJjJk, N(H)C(=NH)NJjJk, N(H)C(=O)NJjJk or N(H)C(=S)NJjJk; or qe and qf together are =C(qg)(qh); qg and qh are each independently H, halogen, C1-C12 alkyl, or substituted C1-C12 alkyl.

[000421] The synthesis and preparation of BNA monomers of methylene-oxy (4'-CH2-O-2') adenine, cytosine, guanine, 5-methylcytosine, thymine, and uracil, along with their oligomerization properties, and nucleic acid recognition have been described (see, for example, Koshkin et al., Tetrahedron, 1998, 54, 3607-3 630). BNAs and preparation thereof are also described in WO 98/39352 and WO 99/14226.

[000422] Methylene-oxy (4'-CH2-O-2') BNA analogs, methylene-oxy (4'-CH2-O-2') BNAs, and 2'-thio-BNAs have also been prepared (see, for example, Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222). The preparation of blocked nucleoside analogues comprising oligodeoxyribonucleotide duplexes as substrates for nucleic acid polymerases has also been described (see, for example, Wengel et al., WO 99/14226). In addition, the synthesis of 2'-amino-BNA, a new behaviorally restricted high-affinity oligonucleotide analogue, has been described in the art (see, for example, Singh et al., J. Org. Chem., 1998, 63, 10035-10039 ). Furthermore, 2'-amino- and 2'-methylamino BNAs were prepared and the thermal stability of their duplexes with RNA and complementary DNA strands were previously reported.

[000423] In certain embodiments, the bicyclic nucleoside having Formula VI: wherein: Bx is a heterocyclic base moiety; Ta and Tb are each independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety, or a covalent bond to a support means; each qi, qj, qk and ql is, independently, H, halogen, C1C12 alkyl, substituted C1-C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkyl, C1C12 alkoxy, substituted C1-C12 alkoxy, OJj, SJj, SOJj, SO2 Jj, NJjJk, N3, CN, C(=O)OJj, C(=O)NJjJk, C(=O)Jj, OC(=O)NJjJk, N(H)C(=NH)NJjJk, N(H)C(=O)NJjJk, or N(H)C(=S)NJjJk; and qi and qj or ql and qk together are =C(qg)(qh), where qg and qh are each independently H, halogen, C1-C12 alkyl, or substituted C1-C12 alkyl.

[000424] A carbocyclic bicyclic nucleoside having a 4'-(CH2)3-2' bridge and the alkenyl manologue, 4'-CH=CH-CH2-2' bridge, has been described (see, for example, Freier et al., Nucleic Acids Research, 1997, 25(22), 4429-4443 and Albaek et al., J. Org. Chem., 2006, 71, 7731-7740). The synthesis and preparation of carbocyclic bicyclic nucleosides along with their oligomerization and biochemical studies have also been described (see, for example, Srivastava et al., J. Am. Chem. Soc. 2007, 129(26), 8362-8379 ).

[000425] As used herein, "bicyclic nucleoside" refers to a nucleoside comprising a bridge connecting two sugar ring carbon atoms, thereby forming a bicyclic sugar moiety. In certain embodiments, a bridge connects the 2' carbon and another carbon on the sugar ring.

[000426] As used herein, "4'-2' bicyclic nucleoside" or "4' to 2' bicyclic nucleoside" refers to a bicyclic nucleoside comprising a furanose ring comprising a bridge connecting the 2' carbon atom and the 4' carbon atom.

[000427] As used herein, "monocyclic nucleosides" refer to nucleosides comprising modified sugar moieties that are not bicyclic sugar moieties. In certain embodiments, the sugar moiety, or sugar moiety analogue, of a nucleoside can be modified or substituted at any position.

[000428] As used herein, "modified 2' sugar" means a furanosyl sugar modified at the 2' position. In certain embodiments, such modifications include substituents selected from: a halide, including, but not limited to, substituted and unsubstituted alkoxy, substituted and unsubstituted thioalkyl, substituted and unsubstituted amino alkyl, substituted and unsubstituted alkyl, substituted and unsubstituted allyl, and substituted and unsubstituted alkynyl. In certain embodiments, the 2' modifications are selected from substituents including, but not limited to: O[(CH2)nO]mCH3, O(CH2)nNH2, O(CH2)nCH3, O(CH2)nONH2, OCH2C(=O)N(H)CH3, and O(CH2)nON[(CH2)nCH3]2, where n and m are from 1 to about 10. Other 2'-substituent groups may also be selected from: C1-C12 alkyl; substituted alkyl; alkenyl; alkynyl; alkaryl; aralkyl; O-alkaryl or O-aralkyl; SH; SCH3; OCN; Cl; Br; CN; CF3; OCF3; SOCH3; SO2CH3; ONO2; NO2; N3; NH2; heterocycloalkyl; heterocycloalkaryl; aminoalkylamino; polyalkylamino; substituted silyl; an RNA cleavage group; a reporter group; an interleaver; a group for improving pharmacokinetic properties; and a group for improving the pharmacokinetic properties of an antisense compound, and other substituents having similar properties. In certain embodiments, the modified nucleosides comprise a 2'-MOE side chain (see, for example, Baker et al., J. Biol. Chem., 1997, 272, 11944-12000). Such a 2'-MOE substitution has been reported to have improved binding affinity compared to non-modified nucleosides and to other modified nucleosides, such as 2'-O-methyl, O-propyl, and O-aminopropyl. Oligonucleotides bearing the 2'-MOE substituent have also been shown to be antisense inhibitors of gene expression with promising characteristics for in vivo use (see, for example, Martin, P., Helv. Chim. Acta, 1995, 78, 486-504; Altmann et al., Chimia, 1996, 50, 168176; Altmann et al., Biochem. Soc. Trans. , 1996, 24, 630-637 and Altmann et al., Nucleosides Nucleotides, 1997, 16, 917-926).

[000429] As used herein, a "modified tetrahydropyran nucleoside" or "modified THP nucleoside" means a nucleoside having a six-membered tetrahydropyran "sugar" replaced by the pentofuranosyl residue in normal nucleosides (a sugar substitute). Modified THP nucleosides include, but are not limited to, what are referred to in the art as hexitol nucleic acid (HNA), anitol nucleic acid (ANA), mannitol nucleic acid (MNA) (see, Leumann, CJ. Bioorg. & Med. Chem. (2002) 10:841-854), fluorine HNA (F-HNA), or those compounds having Formula X: Formula X: wherein independently for each of the at least one tetrahydropyran nucleoside analogue of Formula X: Bx is a heterocyclic base moiety; T3 and T4 are each independently an internucleoside linker linking the tetrahydropyran nucleoside analogue to the antisense compound or one of T3 and T4 is an internucleoside linker linking the tetrahydropyran nucleoside analogue to the antisense compound and the other of T3 and T4 is H, a hydroxyl protecting group, a linked conjugate group, or a 5' or 3' linked terminal group; q1, q2, q3, q4, q5, q6, and q7 are each independently H, C1-C6 alkyl, substituted C1-C6 alkyl, C2-C6 alkenyl, substituted C2-C6 alkenyl, C2-C6 alkynyl, or substituted C2-C6 alkynyl; and one of R1 and R2 is hydrogen and the other is selected from halogen, substituted or unsubstituted alkoxy, NJ1J2, SJ1, N3, OC(=X)J1, OC(=X)NJ1J2, NJ3C(=X)NJ1J2, and CN, where X is O, S, or NJ1, and each J1, J2, and J3 is, independently, H or C1-C6 alkyl.

[000430] In certain embodiments, modified THP nucleosides of Formula X are provided wherein qm, qn, qp, qr, qs, qt, and qu are each H. In certain embodiments, at least one of qm, qn, qp, qr, qs, qt, and qu is other than H. In certain embodiments, at least one of qm, qn, qp, qr, qs, qt, and qu is methyl. In certain embodiments, THP nucleosides of Formula X are provided wherein one of R1 and R2 is F. In certain embodiments, R1 is fluoro and R2 is H, R1 is methoxy and R2 is H, and R1 is methoxyethoxy and R2 is H.

[000431] As used herein, "2'-modified nucleoside" or "2'-substituted nucleoside" refers to a nucleoside comprising a sugar comprising a substituent at the 2' position of a furanose ring other than H or OH. 2'-modified nucleosides include, but are not limited to, bicyclic nucleosides in which the bridge connecting two sugar ring carbon atoms connects the 2' carbon and one other sugar ring carbon, and nucleosides with non-bridge 2' substituents, such as allyl, amino, azido, thio, O-allyl, O-C1-C10 alkyl, -OCF3, O-(CH2)2-O-CH3, 2' -O(CH2)2SCH3, O-(CH2)2-O-N(Rm)(Rn), or O-CH2-C(=O)-N(Rm)(Rn), where each Rm and Rn is independently H or substituted or unsubstituted C1C10 alkyl. 2'-Modified nucleosides may comprise other modifications, for example at other sugar positions and/or at the nucleobase.

[000432] As used herein, "2'-F" refers to a sugar comprising a fluoro group at the 2' position.

[000433] As used herein, "2'-OMe" or "2'-OCH3" or "2'-O-methyl", each, refers to a nucleoside comprising a sugar comprising a -OCH3 group at the 2' position of the sugar ring.

[000434] As used herein, "oligonucleotide" refers to a compound comprising a plurality of linked nucleosides. In certain embodiments, one or more of the plurality of nucleosides is modified. In certain embodiments, an oligonucleotide comprises one or more ribonucleosides (RNA) and/or deoxyribonucleosides (DNA).

[000435] Many other bicyclic and tricyclic sugar substitute ring systems are also known in the art which can be used to modify nucleosides for incorporation into antisense compounds (see, for example, review article: Leumann, J.C., Bioorganic & Medicinal Chemistry, 2002, 10, 841-854). Such ring systems can undergo several additional substitutions to enhance activity.

[000436] Methods for preparing modified sugars are well known to those skilled in the art.

[000437] In nucleotides having modified sugar moieties, the nucleobase moieties (natural, modified, or a combination thereof) are retained for hybridization with an appropriate target nucleic acid.

[000438] In certain embodiments, antisense compounds comprise one or more nucleotides having modified sugar moieties. In certain embodiments, the modified sugar moiety is 2'-MOE. In certain embodiments, the modified 2'-MOE nucleotides are arranged in a gapmer motif. In certain embodiments, the modified sugar moiety is a cEt. In certain embodiments, the modified cEt nucleotides are arrayed through wings of a gapmer motif.

Compositions and Methods for Formulating Pharmaceutical Compositions

[000439] Antisense oligonucleotides can be mixed with pharmaceutically acceptable active or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for formulating pharmaceutical compositions are dependent on a number of criteria, including, but not limited to, the route of administration, the extent of the disease, or what dose is to be administered.

[000440] An antisense compound targeted to an HBV nucleic acid can be used in pharmaceutical compositions by combining the antisense compound with a suitable pharmaceutically acceptable diluent or carrier. A pharmaceutically acceptable diluent includes phosphate buffered saline (PBS). PBS is a suitable diluent for use in compositions to be delivered parenterally. Thus, in one embodiment, employed in the methods described herein is a pharmaceutical composition comprising an antisense compound targeting an HBV nucleic acid and a pharmaceutically acceptable diluent. In certain embodiments, the pharmaceutically acceptable diluent is PBS. In certain embodiments, the antisense compound is an antisense oligonucleotide.

[000441] The pharmaceutical compositions comprising the antisense compounds encompass any pharmaceutically acceptable salts, esters, or salts of such esters, or any other oligonucleotides that, after administration to an animal, including a human being, are able to provide (directly or indirectly) the biologically active metabolite or residue thereof. Thus, for example, the disclosure is also directed to pharmaceutically acceptable salts of antisense compounds, prodrugs, pharmaceutically acceptable salts of such prodrugs, and other bioequivalents. Pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.

[000442] A prodrug may include the incorporation of additional nucleosides at one or both ends of an antisense compound, which are cleaved by endogenous nucleases within the body, to form the active antisense compound.

Conjugated Antisense Compounds

[000443] The antisense compounds can be covalently linked to one or more moieties or conjugates that enhance the activity, cellular distribution or cellular uptake of the resulting antisense oligonucleotides. Typical conjugated groups include cholesterol moieties and lipid moieties. Additional conjugated groups include carbohydrates, phospholipids, biotin, phenazine, folate, phenanthridine, anthraquinone, acridine, fluoresceins, rhodamines, coumarins, and dyes.

[000444] Antisense compounds can also be modified to have one or more stabilizing groups that are usually associated with one or both termini of antisense compounds to enhance properties such as, for example, nuclease stability. Included in stabilization groups are capping (levelling) structures. Such terminal modifications protect the antisense compound having terminal nucleic acid from exonuclease degradation, and may aid in distribution and/or localization within a cell. The capping can be present at the 5'-terminus (5'-capping), or at the 3'-terminus (3'-capping), or it can be present at both ends. Cap structures are well known in the art and include, for example, inverted deoxy abasic caps. Other 3' and 5' stabilizing groups that can be used to flatten one or both ends of an antisense compound to impart nuclease stability include those disclosed in WO 03/004602 published January 16, 2003.

Cell Culture Treatments and Antisense Compounds

[000445] The effects of antisense compounds on the level, activity or expression of HBV nucleic acids can be tested in vitro, in a variety of cell types. Cell types used for such analyzes are available from commercial suppliers (e.g., American Type Culture Collection, Manassus, VA; Zen-Bio, Inc., Research Triangle Park, NC; Clonetics Corporation, Walkersville, MD) and were cultured according to supplier instructions using commercially available reagents (e.g., Invitrogen Life Technologies, Carlsbad, CA). Illustrative cell types include, but are not limited to, cells, HuVEC cells, b.END cells, HepG2 cells, Hep3B cells, and primary hepatocytes.

In vitro test of antisense oligonucleotides

[000446] Described herein are methods for treating cells with antisense oligonucleotides that may be appropriately modified for treatment with other antisense compounds.

[000447] Cells can be treated with antisense oligonucleotides when the cells reach about 60-80% confluency in culture.

[000448] A reagent commonly used to introduce antisense oligonucleotides into cultured cells includes LIPOFECTIN cationic lipid transfection reagent (Invitrogen, Carlsbad, CA). Antisense oligonucleotides can be mixed with LIPOFECTIN in OPTI-MEM 1 (Invitrogen, Carlsbad, CA) to achieve the desired final concentration of antisense oligonucleotide and a LIPOFECTIN concentration that can range from 2 to 12 ug/ml per 100 nM of antisense oligonucleotide.

[000449] Another reagent used to introduce antisense oligonucleotides into cultured cells includes LIPOFECTAMINE (Invitrogen, Carlsbad, CA). The antisense oligonucleotide is mixed with LIPOFECTAMINE in OPTI-MEM 1 reduced serum medium (Invitrogen, Carlsbad, CA) to obtain the desired concentration of antisense oligonucleotide and a concentration of LIPOFECTAMINE that can vary between 2 and 12 ug/ml per 100 nM of antisense oligonucleotide.

[000450] Another technique used to introduce antisense oligonucleotides into cultured cells includes electroporation.

[000451] Cells are treated with antisense oligonucleotides by routine methods. Cells can be harvested 16-24 hours after antisense oligonucleotide treatment, at which time RNA or target nucleic acid protein levels are measured by methods known in the art and described herein. In general, when treatments are performed in multiple replicates, data are presented as the mean of replicate treatments.

[000452] The concentration of antisense oligonucleotide used varies from cell line to cell line. Methods for determining the optimal concentration of antisense oligonucleotides for a particular cell line are well known in the art. Antisense oligonucleotides are normally used at concentrations ranging from 1 to 300 nM when transfected with LIPOFECTAMINE. Antisense oligonucleotides are used at higher concentrations ranging from 625 to 20,000 nM when transfected using electroporation.

RNA isolation

[000453] RNA analysis can be performed on total cellular RNA or poly(A)+ mRNA. RNA isolation methods are well known in the art. RNA is prepared using methods well known in the art, for example, using TRIZOL reagent (Invitrogen, Carlsbad, CA) according to the manufacturer's recommended protocols.

Inhibition analysis of expressions or target levels

[000454] Inhibition of HBV expression or nucleic acid levels can be assessed in a variety of ways known in the art. For example, target nucleic acid levels can be quantified, for example, by Northern blot analysis, competitive polymerase chain reaction (PCR) or real-time quantitative PCR. RNA analysis can be performed on total cellular RNA or poly(A) + mRNA. RNA isolation methods are well known in the art. Northern blot analysis is also routine in the art. Quantitative real-time PCR can be conveniently performed using the ABI PRISM 7600, 7700, or 7900 Sequence Detection System, commercially available from PE-Applied Biosystems, Foster City, CA and used according to the manufacturer's instructions.

Real Time Quantitative PCR Analysis of Target RNA Levels

[000455] Quantitation of target RNA levels can be performed by real-time quantitative PCR using an ABI Prism 7600, 7700, or 7900 Sequence Detection System (PE-Applied Biosystems, Foster City, CA) according to the manufacturer's instructions. Quantitative real-time PCR methods are well known in the art.

[000456] Prior to real-time PCR, the isolated RNA is subjected to a reverse transcriptase (RT) reaction, which produces complementary DNA (cDNA), which is then used as a substrate for real-time PCR. Real-time PCR and RT reactions are performed sequentially on the same sample. Real-time PCR and RT reactions can be obtained from Invitrogen (Carlsbad, CA). RT real-time PCR reactions are performed by methods well known to those skilled in the art.

[000457] The amounts of target gene (or RNA) obtained by real-time PCR are normalized using the expression level of a gene whose expression is constant, such as cyclophilin A or by quantification of total RNA using RIBOGREEN (Invitrogen, Inc., Carlsbad, CA). Cyclophilin A expression is quantified by real-time PCR, running simultaneously with the target, multiplexing, or separately. Total RNA was quantitated using RIBOGREEN RNA Quantitation Reagent (Invetrogen, Inc., Eugene, OR). RNA quantification methods by RIBOGREEN RNA are taught in Jones, L.J., et al, (Analytical Biochemistry, 1998, 265, 368-374). A CYTOFLUOR 4000 instrument (PE Applied Biosystems) is used to measure RIBOGREEN fluorescence.

[000458] Probes and primers are designed to hybridize with an HBV nucleic acid. Methods for designing real-time PCR probes and primers are well known in the art and may include the use of software such as PRIMER EXPRESS Software (Applied Biosystems, Foster City, CA).

Real Time Quantitative PCR Analysis of Target DNA Levels

[000459] Quantitation of target DNA levels can be performed by real-time quantitative PCR using the ABI PRISM 7600, 7700, or 7900 Sequence Detection System (PE-Applied Biosystems, Foster City, CA) according to the manufacturer's instructions. Quantitative real-time PCR methods are well known in the art.

[000460] Target gene (or DNA) amounts obtained by real-time PCR are normalized using the expression level of a gene whose expression is constant, such as cyclophilin A, or by quantification of total DNA using RIBOGREEN (Invitrogen, Inc., Carlsbad, CA). Cyclophilin A expression is quantified by real-time PCR, running simultaneously with the target, multiplexing, or separately. Total DNA is quantitated using RIBOGREEN RNA Quantitation Reagent (Invetrogen, Inc., Eugene, OR). RIBOGREEN DNA quantification methods are taught in Jones, L.J., et al, (Analytical Biochemistry, 1998, 265, 368-374). A CYTOFLUOR 4000 instrument (PE Applied Biosystems) is used to measure RIBOGREEN fluorescence.

[000461] Probes and primers are designed to hybridize with an HBV nucleic acid. Methods for designing real-time PCR probes and primers are well known in the art and may include the use of software such as PRIMER EXPRESS Software (Applied Biosystems, Foster City, CA).

Protein Level Analysis

[000462] Antisense inhibition of HBV nucleic acids can be determined by measuring HBV protein levels. HBV protein levels can be assessed or quantitated in a variety of ways well known in the art, such as immunoprecipitation, Western blot analysis (immunoblotting), enzyme-linked immunosorbent assay (ELISA), quantitative protein assays, protein activity assays (e.g., caspase activity assays), immunohistochemistry, immunocytochemistry, or fluorescence-activated cell sorting (FACS). Targeted antibodies can be identified and obtained from a variety of sources, such as the MSRS antibody catalog (Aerie Corporation, Birmingham, MI), or can be prepared by conventional methods of generating monoclonal or polyclonal antibodies well known in the art.

In vivo testing of antisense compounds

[000463] Antisense compounds, for example, antisense oligonucleotides, are tested in animals to assess their ability to inhibit HBV expression and produce phenotypic changes. The tests can be performed in normal animals, or in experimental models of the disease. For administration to animals, the antisense oligonucleotides are formulated in a pharmaceutically acceptable diluent, such as phosphate buffered saline. Administration includes parenteral routes of administration, such as intraperitoneal, intravenous, subcutaneous, intrathecal, and intracerebroventricular. Calculation of antisense oligonucleotide dosing and dosing frequency is well within the capabilities of those skilled in the art, and is dependent on factors such as the route of administration and the body weight of the animal. After a period of treatment with antisense oligonucleotides, RNA is isolated from liver tissue and changes in HBV nucleic acid expression are measured. Changes in HBV DNA levels are also measured. Changes in HBV protein levels are also measured. Changes in HBV HBeAg levels are also measured. Changes in HBV HBsAg levels are also measured.

Certain Indications

[000464] In certain embodiments, methods, compounds, and compositions for treating a subject comprising administering one or more pharmaceutical compositions provided herein are provided herein. In certain embodiments, the subject has an HBV-related condition. In certain embodiments, chronic HBV infection, inflammation, fibrosis, cirrhosis, liver cancer, serum hepatitis, jaundice, liver cancer, liver inflammation, liver fibrosis, liver cirrhosis, liver failure, diffuse hepatocellular inflammatory disease, hemophagocytic syndrome, serum hepatitis, and HBV viremia. In certain embodiments, the HBV-related condition may have what may include any or all of the following: flu-like illness, weakness, aches and pains, headache, fever, loss of appetite, diarrhea, jaundice, nausea and vomiting, pain over the liver area of the body, clay-colored or gray stools, itching all over the body, and dark-colored urine, when combined with a positive test for the presence of a hepatitis B virus, a hepatitis B viral antigen, or a positive test for the presence of an antibody specific for a hepatitis B viral antigen. In certain embodiments, the subject is at risk for an HBV-related condition. This includes people with one or more risk factors for developing an HBV-related condition, including sexual exposure to an individual infected with the hepatitis B virus, living in the same household as an individual with a lifetime infection with the hepatitis B virus, exposure to human blood infected with the hepatitis B virus, injecting illicit drugs, being a person who has hemophilia, and visiting an area where hepatitis B is common. In certain embodiments, the individual has been identified as in need of treatment for an HBV-related condition. In certain embodiments methods for prophylactically reducing HBV expression in an individual are provided herein. Certain embodiments include treating a subject in need thereof by administering to the subject a therapeutically effective amount of an antisense compound targeted to an HBV nucleic acid.

[000465] Due to overlapping transmission routes, many people have been exposed to both hepatitis B virus (HBV) and hepatitis C virus (HCV), and a smaller proportion are chronically infected with both viruses, especially in regions such as Asia where HBV is endemic. Estimates suggest that up to 10% of people with HCV may also have HBV, while perhaps 20% of people with HBV are co-infected with HCV. However, the treatment of hepatitis B or hepatitis B in individuals co-infected with HBV-HCV has not been well studied. Treatment is complicated by the fact that HCV and HBV appear to inhibit each other's replication (although not all studies have observed this interaction). Thus, treatment that completely suppresses HBV could potentially allow HCV to re-emerge, or vice versa. Therefore, the compounds and compositions described herein can be advantageously used for the treatment of patients infected with HBV and HCV. Examples of treatment options for hepatitis C (HCV) include interferons, eg interferon alfa-2b, interferon alfa-2a and interferon alfacon-1. Less frequent interferon dosing can be achieved using pegylated interferon (interferon bound to a polyethylene glycol moiety which improves its pharmacokinetic profile). Combination therapy with interferon alfa-2b (pegylated and non-pegylated) and ribavirin has also been shown to be effective for some patient populations. Other agents currently being developed include inhibitors of HCV RNA replication (eg, ViroPharma's VP50406 series), HCV antisense agents, HCV therapeutic vaccines, HCV protease inhibitors, HCV helicase inhibitors, and HCV antibody therapy (monoclonal or polyclonal).

[000466] In certain embodiments, treatment with the methods, compounds, and compositions described herein are useful for preventing an HBV-related condition associated with the presence of the hepatitis B virus. In certain embodiments, treatment with the methods, compounds, and compositions described herein are useful for preventing an HBV-related condition.

[000467] In one embodiment, administering a therapeutically effective amount of an antisense compound targeted to an HBV nucleic acid is accompanied by monitoring HBV mRNA levels in a subject's serum to determine a subject's response to administration of the antisense compound. In certain embodiments, administration of a therapeutically effective amount of an antisense compound targeted to an HBV nucleic acid is accompanied by monitoring levels of HBV DNA in a subject's serum to determine the subject's response to administration of the antisense compound. In certain embodiments, administration of a therapeutically effective amount of an antisense compound targeted to an HBV nucleic acid is accompanied by monitoring levels of HBV protein in a subject's serum to determine the subject's response to administration of the antisense compound. In certain embodiments, administration of a therapeutically effective amount of an antisense compound targeted to an HBV nucleic acid is accompanied by monitoring levels of HBV S antigens (HBsAg) in a subject's serum to determine the subject's response to administration of the antisense compound. In certain embodiments, administration of a therapeutically effective amount of an antisense compound targeted to an HBV nucleic acid is accompanied by monitoring levels of HBV E antigen (HBeAg) in a subject's serum to determine the subject's response to administration of the antisense compound. An individual's response to administration of the antisense compound is used by a physician to determine the amount and duration of therapeutic intervention.

[000468] In certain embodiments, administration of an antisense compound targeted to an HBV nucleic acid results in the reduction of HBV expression by at least 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 99%, or a range defined by two of these values. In certain embodiments, administration of an antisense compound targeted to an HBV nucleic acid results in reduction of symptoms associated with the HBV-related condition and reduction of HBV-related markers in the blood. In certain embodiments, administration of an HBV antisense compound decreases HBV RNA levels, HBV DNA levels, HBV protein levels, HBsAg levels, or HBeAg levels by at least 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85 , 90, 95 or 99%, or a range defined by two of these values.

[000469] In certain embodiments, pharmaceutical compositions comprising an antisense compound targeted to HBV are used for the preparation of a medicament to treat a patient suffering from, or susceptible to, an HBV-related condition.

Certain Combination Therapies

[000470] In certain embodiments, one or more pharmaceutical compositions provided herein are co-administered with one or more other pharmaceutical agents. In certain embodiments, such one or more other pharmaceutical agents are intended to treat the same disease, disorder or condition as one or more pharmaceutical compositions provided herein. In certain embodiments, such one or more other pharmaceutical agents are intended to treat a different disease, disorder or condition as one or more pharmaceutical compositions provided herein. In certain embodiments, such one or more other pharmaceutical agents are intended to treat an undesirable side effect of one or more pharmaceutical compositions provided herein. In certain embodiments, one or more pharmaceutical compositions provided herein are co-administered with another pharmaceutical agent to treat an undesired effect of another pharmaceutical agent. In certain embodiments, one or more pharmaceutical compositions provided herein are co-administered with another pharmaceutical agent to produce a combinatorial effect. In certain embodiments, one or more pharmaceutical compositions provided herein are co-administered with another pharmaceutical agent to produce a synergistic effect.

[000471] In certain embodiments, one or more pharmaceutical compositions provided herein and one or more other pharmaceutical agents are administered at the same time. In certain embodiments, one or more pharmaceutical compositions provided herein and one or more other pharmaceutical agents are administered at different times. In certain embodiments, one or more pharmaceutical compositions provided herein and one or more other pharmaceutical agents are prepared together in a single formulation. In certain embodiments, one or more pharmaceutical compositions provided herein and one or more other pharmaceutical agents are prepared separately. In certain embodiments the disclosed antisense oligonucleotides are administered in combination with an HCV agent. In further embodiments, the HCV compound is administered simultaneously with the antisense compound, in other embodiments, the HCV compound can be administered separately, such that a dose of each of the HCV agent and the antisense compounds overlaps, over time, within the patient's body. In related embodiments, the HCV agent can be selected from interferon alfa-2b, interferon alfa-2a, and interferon alfacon-1 (pegylated and non-pegylated), ribavirin, an inhibitor of HCV RNA replication (e.g., ViroPharma's VP50406 series), an HCV antisense agent, an HCV therapeutic vaccine, an HCV protease inhibitor, an HCV helicase inhibitor, and an HCV antibody therapy. CV (monoclonal or polyclonal).

[000472] In other embodiments, an HBV antisense compound of the present invention can be administered to an HBV-infected patient in combination with one or more HBV therapeutic agents, wherein the one or more HBV therapeutic agents can be administered in the same drug formulation as the HBV ASO compound, or can be administered in a separate formulation. The one or more HBV therapeutic agents can be administered simultaneously with the ASO HBV compound, or they can be administered separately, such that a dose of each of the ASO HBV compound and HBV therapeutic agent overlaps, over time, within the patient's body. In related embodiments, the one or more HBV therapeutic agents can be selected from interferon alfa-2b, interferon alfa-2a and interferon alfacon-1 (pegylated and non-pegylated), ribavirin, an inhibitor of HBV RNA replication, a second HBV antisense compound; a therapeutic HBV vaccine; a prophylactic HBV vaccine; lamivudine (3TC); entecavir, tenofovir; telbivudine (LdT); adefovir, and an HBV antibody therapy (monoclonal or polyclonal).

EXAMPLES Non-limiting disclosure and incorporation by reference

[000473] While certain compounds, compositions, and methods described herein have been specifically described in accordance with certain embodiments, the examples that follow serve only to illustrate the compounds described herein and are not intended to be limiting thereof. Each of the references referred to in the present application is hereby incorporated by reference in its entirety.

Example 1: Antisense Inhibition of HBV viral mRNA in HepG2.2.15 cells by MOE gapmers

[000474] Antisense oligonucleotides were designed to target an HBV viral nucleic acid and were tested for their effects on HBV mRNA in vitro. HepG2.2.15 cells grown at a density of 25,000 cells per well were transfected using electroporation with 15,000 nM antisense oligonucleotide. After a period of approximately 24 hours of treatment, RNA was isolated from the cells and HBV mRNA levels were measured by quantitative real-time PCR. The defined viral primer probe RTS3370 (forward sequence CTTGGTCATGGGCCATCAG, designated here as SEQ ID NO: 2; reverse sequence CGGCTAGGAGTTCCGCAGTA, designated here as SEQ ID NO: 3; probe sequence TGCGTGGAACCTTTTCGGCTCC, designated here as SEQ ID NO: 4) was used to measure mRNA levels. HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. Results are presented as percentage of HBV inhibition, relative to untreated control cells.

[000475] The novel chimeric antisense oligonucleotides designated in Table 1 were designated as either 5-10-5 MOE gapmers or 3-10-4 MOE gapmers. 5-10-5 MOE gapmers are 20 nucleosides long, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising five nucleosides each. 3-10-4 MOE gapmers are 17 nucleosides long, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising three and 4 nucleosides respectively. Each nucleoside in the 5' wing segment and each nucleoside in the 3' wing segment has an MOE sugar modification. Each nucleoside in the center gap segment has a deoxy sugar modification. The internucleoside linkages across each gapmer are phosphorothioate linkages (P=S). All cytosine residues across each gapmer are 5-methylcytosines.

[000476] "Viral target starting site" indicates the maximum 5' nucleotide to which the gapmer is targeted in the viral gene sequence. "Viral target stop site" indicates the 3'-maximum nucleotide at which the gapmer is targeted in the viral gene sequence. The 'Reason' column indicates the gap and wing structure of each gapmer. Each gapmer listed in Table 1 is a target for the viral genomic sequence, designated here as SEQ ID NO: 1 (GENBANK Accession No. U95551.1). Table 1 Inhibition of HBV viral mRNA levels by MOE gapmers targeted to SEQ ID NO: 1

Example 2: Antisense inhibition of HBV viral mRNA in HepG2.2.15 cells by MOE gapmers

[000477] Additional antisense oligonucleotides were designed targeting an HBV viral nucleic acid and were tested for their effects on HBV mRNA in vitro. HepG2.2.15 cells grown at a density of 25000 cells per well were transfected using electroporation with 15000 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from cells and HBV mRNA levels were measured by quantitative real-time PCR. The RTS3370 primer viral probe setup was used to measure mRNA levels. RTS3370 detects full-length mRNA and second portions of pre-S1, pre-S2, and pre-C mRNA transcripts. The gapmers were also probed with additional primer probe configurations. The RTS3371 viral probe setup (forward sequence CCAAACCTTCGGACGGAAA, designated here as SEQ ID NO: 311; reverse sequence TGAGGCCCACTCCCATAGG, designated here as SEQ ID NO: 312; probe sequence CCCATCATCCTGGGCTTTCGGAAAAT, designated here as SEQ ID NO: 313) was also used to measure mRNA levels. RTS3371 detects full length mRNA and second portions of pre-S1, pre-S2 and pre-C mRNA transcripts, similar to RTS3370 but in different regions. The RTS3372 viral probe setup (forward sequence ATCCTATCAACACTTCCGGAAACT, designated here as SEQ ID NO: 314; reverse sequence CGACGCGGCGATTGAG, designated here as SEQ ID NO: 315; probe sequence AAGAACTCCCTCGCCTCGCAGACG, designated here as SEQ ID NO: 316) was used to measure mRNA levels. RTS3372 detects the full-length genomic sequence. The RTS3373MGB viral probe setup (forward sequence CCGACCTTGAGGCATACTTCA, designated here as SEQ ID NO: 317; reverse sequence AATTTATGCCTACAGCCTCCTAGTACA, designated here as SEQ ID NO: 318; probe sequence TTAAAGACTGGGAGGAGTTG, designated here as SEQ ID NO: 319) was used to measure mRNA levels. RTS3373MGB detects full-length mRNA and second portions of pre-S1, pre-S2, pre-C, and pre-X mRNA transcripts.

[000478] HBV mRNA levels were adjusted according to total RNA content as measured by RIBOGREEN®. Results are presented as percentage of HBV inhibition, relative to untreated control cells.

[000479] The novel chimeric antisense oligonucleotides designated in Table 2 were designated as either 5-10-5 MOE gapmers, 3-10-3 MOE gapmers, or 2-10-2 MOE gapmers. 510-5 MOE gapmers are 20 nucleosides long, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising five nucleosides each. 3-10-3 MOE gapmers are 16 nucleosides long, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising three nucleosides each. The 2-10-2 MOE gapmers are 14 nucleosides in length, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising two nucleosides each. Each nucleoside in the 5' wing segment and each nucleoside in the 3' wing segment has an MOE sugar modification. Each nucleoside in the center gap segment has a deoxy sugar modification. The internucleoside linkages across each gapmer are phosphorothioate linkages (P=S). All cytosine residues across each gapmer are 5'-methylcytosines.

[000480] "Start site" indicates the 5'-maximum nucleotide at which the gapmer is targeted in the viral gene sequence. "Stop site" indicates the maximum 3'-nucleotide at which the gapmer is targeted in the viral gene sequence. The 'Reason' column indicates the gap and wing structure of each gapmer. Each gapmer listed in Table 2 is a target for the viral genomic sequence, designated here as SEQ ID NO: 1 (GENBANK Accession No. U95551.1). Table 2 Inhibition of HBV viral mRNA levels by MOE gapmers targeted to SEQ ID NO: 1 (detected by RTS3370, RTS3371, RTS3372 and RTS3373MGB)

Example 3: Antisense inhibition of HBV viral mRNA in HepAD38 (Tet-HBV) cells by MOE gapmers

[000481] Certain antisense oligonucleotides selected from the study described in Example 2 were tested for their effects on HBV mRNA in another cell line, HepAD38 human hepatoma cells, in which HBV production is under the control of a tetracycline-regulated promoter. HepAD38 (Tet-HBV) cells grown at a density of 45,000 cells per well were transfected using electroporation with 15,000 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from cells and HBV mRNA levels were measured by quantitative real-time PCR. RTS3372 and RTS3373MGB viral primer probe configurations were used individually to measure mRNA levels. HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. The results are shown in Table 3 as percent inhibition of HBV, relative to untreated control cells. Table 3 Inhibition of HBV viral mRNA levels by MOE gapmers in HepAD38 (Tet-HBV) cells (detected by RTS3372 and RTS3373MGB)

Example 4: Antisense inhibition of HBV viral mRNA in HepAD38 (Tet-HBV) cells by MOE gapmers

[000482] Certain antisense oligonucleotides from the study described in Examples 1 and 2 were tested for their effects on HBV mRNA in vitro. HepAD38 (Tet-HBV) cells grown at a density of 45,000 cells per well were transfected using electroporation with 15,000 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from cells and HBV mRNA levels were measured by quantitative real-time PCR. The RTS3372 primer viral probe setup was used to measure mRNA levels. mRNA levels were also measured using the RTS3373MGB primer setup. HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. The results are shown in Table 4 as percent inhibition of HBV, relative to untreated control cells. Table 4 Inhibition of HBV viral mRNA levels by MOE gapmers (RTS3372 and RTS3373MGB)

Example 5: Dose-dependent inhibition of viral HBV RNA in HepG2.2.15 cells by MOE gapmers

[000483] Certain gapmers from the study described in Examples 3 and 4 were tested at various doses on human HepG2.2.15 cells. Cells were seeded at a density of 25,000 cells per well and transfected using electroporation with concentrations of 2.5 µM, 5.0 µM, 10.0 µM, and 20.0 µM antisense oligonucleotide, as specified in Table 5. After the approximately 16 hour treatment period, RNA was isolated from the cells and HBV mRNA levels were measured by quantitative real-time PCR. The RTS3370 primer viral probe setup was used to measure mRNA levels. HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. Results are presented as percentage of HBV inhibition, relative to untreated control cells.

[000484] The half maximal inhibitory concentration (IC50) of each oligonucleotide is also shown in Table 5. As illustrated in Table 5, HBV mRNA levels were significantly reduced in a dose-dependent manner in cells treated with antisense oligonucleotide. Table 5 Dose-dependent antisense inhibition of HBV RNA in HepG2.2.15 cells using RTS3370

Example 6: Dose dependent inhibition of viral HBV RNA in HepG2.2.15 cells by MOE gapmers

[000485] Additional Gapmers from the study described in Examples 3 and 4 were further tested at various doses on human HepG2.2.15 cells. Cells were seeded at a density of 28,000 cells per well and transfected using LipofectAMINE 2000® reagent with concentrations of 15.625 nM, 31.25 nM, 62.5 nM, 125.0 nM, and 250.0 nM of antisense oligonucleotide, as specified in Table 6. After the treatment period of approximately 16 hours, RNA was isolated from the cells and the HBV mRNA levels were measured by quantitative real-time PCR. The RTS3370 primer viral probe setup was used to measure mRNA levels. HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. Results are presented as percentage of HBV inhibition, relative to untreated control cells.

[000486] The half maximal inhibitory concentration (IC50) of each oligonucleotide is also shown in Table 6. As illustrated in Table 6, HBV mRNA levels were significantly reduced in a dose-dependent manner in some cells treated with antisense oligonucleotide. Table 6 Dose-dependent antisense inhibition of HBV RNA in cells

[000487] mRNA levels were also measured with RTS3371 primer probe setup. The results are shown in Table 7. Table 7 Dose-dependent antisense inhibition of HBV RNA in HepG2.2.15 cells using RTS3371

Example 7: Tolerability of MOE gapmers targeting HBV in BALB/c mice

[000488] BALB/c mice (Charles River, MA) are a multipurpose mouse model often used for safety and efficacy testing. Mice were treated with ISIS antisense oligonucleotides selected from the studies described above and evaluated for changes in levels of various metabolic markers.

Study 1

[000489] Groups of four BALB/c mice each were injected subcutaneously twice a week for 3 weeks with 50 mg/kg of ISIS 146779, ISIS 146786, ISIS 505317, ISIS 505319, ISIS 505330, ISIS 505332, ISIS 505339, ISIS 50 A group of four mice s BALB/c was injected subcutaneously twice a week for 3 weeks with 50 mg/kg of ISIS 141923 (CCTTCCCTGAAGGTTCCTCC (SEQ ID NO: 320)), a 5-10-5 MOE gapmer with no known homology to any human or mouse sequence. Another group of 4 BALB/c mice were injected subcutaneously twice a week for 3 weeks with PBS. This control group served as the control group. Three days after the last dose at each time point, body weights were taken, mice were euthanized, and organs and plasma were harvested for further analysis.

Organ and body weights

[000490] The body weights of the mice were measured pre-dose and at the end of each treatment period. Body weights are shown in Table 8, and are expressed as a percentage change from the weight taken before starting treatment. Liver, spleen, and kidney weights were measured at the end of the study, and are shown in Table 9, as a percentage difference from the respective organ weights of the PBS control. The results indicate that most ISIS oligonucleotides do not cause any adverse effects on organ and body weights. Table 8 Changes in body weights of BALB/c mice after treatment with antisense oligonucleotide (%) Table 9 Changes in organ weights of BALB/c mice after treatment with antisense oligonucleotide (%)

Liver Function

[000491] To assess the effect of ISIS oligonucleotides on liver function, plasma transaminase concentrations were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). Plasma ALT (alanine transaminase) and AST (aspartate transaminase) concentrations were measured and the results are shown in Table 10 expressed as IU/L. Plasma cholesterol and triglyceride levels were also measured using the same clinical chemistry analyzer and the results are also shown in Table 10. Table 10 Effect of antisense oligonucleotide treatment on metabolic markers in the liver of BALB/c mice

Kidney function

[000492] To assess the effect of ISIS oligonucleotides on kidney function, plasma concentrations of blood urea nitrogen (BUN) were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are presented in Table 11, expressed in mg/dL. Table 11 Effect of antisense oligonucleotide treatment on kidney markers in BALB/c mice

Study 2

[000493] Groups of four BALB/c mice each were injected subcutaneously twice a week for 3 weeks with 50 mg/kg of ISIS 505329, ISIS 509926, ISIS 509927, ISIS 509958, ISIS 509959, ISIS 509960, ISIS 509974, ISIS 50 9975, ISIS 510038, ISIS 510039, ISIS 510040, ISIS 510041, and ISIS 510050. A group of 4 BALB/c mice were injected subcutaneously twice a week for 3 weeks with PBS. This control group served as the control group. Three days after the last dose at each time point, body weights were taken, mice were euthanized, and organs and plasma were harvested for further analysis.

Organ Weights

[000494] Liver, spleen, and kidney weights were measured at the end of the study, and are also shown in Table 12 as a percentage change over the respective organ weights from the PBS control. Table 12 Changes in organ weights of BALB/c mice after antisense oligonucleotide treatment (%)

Liver Function

[000495] To assess the effect of ISIS oligonucleotides on liver function, plasma concentrations of transaminases were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). Plasma ALT (alanine transaminase) and AST (aspartate transaminase) concentrations were measured and the results are shown in Table 13 expressed as IU/L. Table 13 Effect of antisense oligonucleotide treatment on transaminases (IU/L) in the liver of BALB/c mice

Kidney function

[000496] To assess the effect of ISIS oligonucleotides on kidney function, plasma concentrations of blood urea nitrogen (BUN) were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are presented in Table 14, expressed in mg/dL. Table 14 Effect of antisense oligonucleotide treatment on kidney markers in BALB/c mice

Example 8: Dose-responsive confirmation of MOE gapmers targeting HBV in HepG2.2.15 cells

[000497] Gapmers were chosen on the basis of sequence conservation, activity, and tolerability, as measured in the study described in Examples 7 and 8, and and tested at various doses in HepG2.2.15 cells. Cells were seeded at a density of 28000 cells per well and transfected using LipofectAMINE 2000 reagent with concentrations of 15.625 nM, 31.25 nM, 62.5 nM, 125.0 nM and 250.0 nM antisense oligonucleotide. Two days after transfection, the medium was replaced with fresh medium. Samples were taken 4 days after transfection. DNA, RNA, HBsAg and HBeAg levels were measured in the supernatant.

[000498] HBV mRNA levels were measured by quantitative real-time PCR. RTS3370 HBV primer probe setup was used to measure mRNA levels. HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. Results are presented as percentage of HBV inhibition, relative to untreated control cells. As illustrated in Table 15, HBV mRNA levels were reduced in a dose-dependent manner in most cells treated with antisense oligonucleotide.

[000499] The HBV antigens in the supernatants were detected using the ELISA technique. HBs antigen (HBsAg) levels were detected by ELISA from Abazyme LLC, MA. As shown in Table 16, treatment with ISIS oligonucleotides 146779, 146786, 505329, 505330, 505339, 505347, 505358, 509927, 509934, 509958, 509959, 509960, 509974, 5100038, 510039, 510040, 510041, 510100, 510106, and 510116 caused significant reduction in HBsAg levels. HBe antigen (HBeAg) levels were detected by ELISA from International Immuno-diagnostics, CA. As shown in Table 17, treatment with ISIS oligonucleotides 146779, 146786, 505329, 505330, 505339, 505347, 505358, 509927, 509934, 509958, 509959, 509960, 509974, 5100038, 510039, 510040, 510041, 510100, 510106, and 510116 caused significant reduction in HBeAg levels. HBV DNA levels were measured using RTS3370 primer setup. As shown in Table 18, treatment with ISIS oligonucleotides 146779, 146786, 505329, 505330, 505339, 505347, 505358, 509927, 509934, 509958, 509959, 509960, 509974, 5100038, 510039, 510040, 510041, 510100, 510106, and 510116 caused significant reduction in HBV DNA levels. Total protein in the supernatants as measured by the DC protein assay (BioRad), as shown in Table 19. Table 15 Dose-dependent antisense inhibition of HBV RNA in HepG2.2.15 cells Table 16 Dose-dependent reduction of S antigen in HepG2.2.15 cell supernatant Table 17 Dose-dependent reduction of E antigen in HepG2.2.15 cell supernatant Table 18 Dose-dependent antisense inhibition of HBV DNA in HepG2.2.15 cells Table 19 Total protein levels in HepG2.2.15 cell supernatant

Example 9: In vivo inhibition of HBV mRNA by MOE gapmers in HBV transgenic mice

[000500] ISIS 146786, a 5-10-5 MOE gapmer, and ISIS 510100, a 3-10-4 MOE gapmer, both demonstrating significant inhibition of HBV mRNA, were tested in transgenic mice containing the HBV gene (Chisari 1.3.32 lineage) (Guidotti, L. G. et al., J. Virol. 1 995, 69, 6158-6169) and the effectiveness of gapmers was evaluated.

Treatment

[000501] Two groups of 10-11 male and female HBV transgenic mice each were administered subcutaneously twice a week for four weeks with 25 mg/kg of ISIS 146786 or ISIS 510100. Another group of 14 male and female HBV transgenic mice were administered entecavir, an oral antiviral drug used to treat the infection for hepatitis B, at 1 mg/kg per day for two weeks. Another group of 10 male and female HBV transgenic mice were injected subcutaneously with PBS twice a week for four weeks. Mice injected with PBS served as the control group. Liver DNA and HBV mRNA levels, plasma ALT, and body and organ weights were measured.

RNA analysis

[000502] RNA was extracted from liver tissue for real-time PCR analysis of HBV using primer probe configurations RTS3370, RTS3371, and RTS3372. Results are presented as percentage inhibition of HBV mRNA relative to PBS control. As shown in Table 20, treatment with ISIS antisense oligonucleotides resulted in significant reduction of HBV mRNA compared to PBS control, independent of the primer probe configuration used for measurement. Entecavir did not decrease HBV mRNA expression. Table 20 Inhibition of HBV mRNA in liver HBV transgenic mice versus PBS control

DNA analysis

[000503] DNA was extracted from liver tissue for HBV real-time PCR analysis using RTS3370 and RTS3371 primer probe configurations. Levels were normalized to RIBOGREEN®. Results are presented as percentage inhibition of HBV DNA, relative to the PBS control. As shown in Table 21, treatment with ISIS antisense oligonucleotides resulted in significant reduction of HBV DNA compared to the PBS control, independent of the primer probe configuration used for measurement. Entecavir treatment also reduced DNA levels, as expected. Table 21 Inhibition of HBV DNA in liver HBV transgenic mice versus PBS control

Liver Function

[000504] To assess the effect of ISIS oligonucleotides on liver function, plasma transaminase concentrations were measured using a manual clinical chemistry analyzer (Teco Diagnostics, Anaheim, CA). Plasma ALT (alanine transaminase) concentrations were measured and the results are shown in Table 22, expressed as IU/L. The results indicate that antisense inhibition of HBV had no adverse effects on liver function in mice. Table 22 Effect of antisense oligonucleotide treatment on liver ALT of transgenic mice

[000505] Study data indicated that both ISIS 146786 and ISIS 510100 caused robust reduction in HBV RNA and liver DNA and treatment with these oligonucleotides were well tolerated in the transgenic mice.

Example 10: Antisense Inhibition of HBV viral mRNA in HepG2.2.15 Cells by MOE gapmers

[000506] Additional antisense oligonucleotides were designed targeting an HBV viral nucleic acid and were tested for their effects on HBV mRNA in vitro. Several of the antisense oligonucleotides from the studies described above were also included in the assay. HepG2.2.15 cells grown at a density of 28000 cells per well were transfected using LipofectAMINE 2000® reagent with 100 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from cells and HBV mRNA levels were measured by quantitative real-time PCR. The RTS3370 primer viral probe setup was used to measure mRNA levels. HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. Results are presented as percentage of HBV inhibition, relative to untreated control cells.

[000507] The new chimeric antisense oligonucleotides designated in Table 23 were designated as 5-10-5 MOE gapmers. Gappers are 20 nucleosides long, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising five nucleosides each. Each nucleoside in the 5' wing segment and each nucleoside in the 3' wing segment has an MOE sugar modification. Each nucleoside in the center gap segment has a deoxy sugar modification. The internucleoside linkages across each gapmer are phosphorothioate linkages (P=S). All cytosine residues across each gapmer are 5-methylcytosines.

[000508] "Viral target start site" indicates the 5'-maximum nucleotide to which the gapmer is targeted in the viral gene sequence. "Viral target stop site" indicates the 3'-maximum nucleotide at which the gapmer is targeted in the viral gene sequence. Each gapmer listed in Table 23 is a target for the viral genomic sequence, designated herein as SEQ ID NO: 1 (GENBANK Accession No. U95551.1). Table 23 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides targeted to SEQ ID NO: 1 (RTS3370)

Example 11: Antisense Inhibition of HBV viral mRNA in HepG2.2.15 Cells by MOE gapmers

[000509] Additional antisense oligonucleotides were designed targeting an HBV viral nucleic acid and were tested for their effects on HBV mRNA in vitro. Several of the antisense oligonucleotides from the studies described above were also included in the assay. HepG2.2.15 cells grown at a density of 28,000 cells per well were transfected using LipofectAMINE 2000® reagent with 70 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from cells and HBV mRNA levels were measured by quantitative real-time PCR. The RTS3370 primer viral probe setup was used to measure mRNA levels. The mRNA levels of some of the gapmers were also measured using RTS3372. HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. Results are presented as percentage of HBV inhibition, relative to untreated control cells.

[000510] The new chimeric antisense oligonucleotides designated in Tables 24 and 25 were designated as gapmers 510-5 MOE. Gappers are 20 nucleosides long, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising five nucleosides each. Each nucleoside in the 5' wing segment and each nucleoside in the 3' wing segment has an MOE sugar modification. Each nucleoside in the center gap segment has a deoxy sugar modification. The internucleoside linkages across each gapmer are phosphorothioate linkages (P=S). All cytosine residues across each gapmer are 5-methylcytosines.

[000511] "Viral target starting site" indicates the 5'-maximum nucleotide to which the gapmer is targeted in the viral gene sequence. "Viral target stop site" indicates the 3'-maximum nucleotide at which the gapmer is targeted in the viral gene sequence. Each gapmer listed in Table 24 is a target for the viral genomic sequence, designated herein as SEQ ID NO: 1 (GENBANK Accession No. U95551.1). Each gapmer listed in Table 25 is a target for the viral genomic sequence, designated here as SEQ ID NO: 1286 (a swapped version of GENBANK Accession No. U95551.1). 'n/a' indicates that the inhibition data for the particular gapmer was not measured with the particular primer probe configuration. Table 24 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides targeted to SEQ ID NO: 1 (RTS3370 and RTS3372) Table 25 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides targeted to SEQ ID NO: 1286 (RTS3370 and RTS3372)

Example 12: Dose dependent inhibition of HBV viral RNA in HepG2.2.15 Cells by MOE gapmers

[000512] Certain gapmers from the study described in Examples 11 and 12 were tested at various doses on human HepG2.2.15 cells. Cells were seeded at a density of 28,000 cells per well and transfected using LipofectAMINE 2000® reagent with concentrations of 5.56 nM, 16.67 nM, 50.0 nM, and 150.0 nM antisense oligonucleotide, as specified in Table 26. After the treatment period of approximately 16 hours, RNA was isolated from cells and HBV mRNA levels were measured by quantitative real-time PCR. The RTS3370 primer viral probe setup was used to measure mRNA levels. HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. Results are presented as percentage of HBV inhibition, relative to untreated control cells.

[000513] The half maximal inhibitory concentration (IC50) of each oligonucleotide is also shown in Table 26. As illustrated in Table 26, HBV mRNA levels were reduced in a dose-dependent manner in cells treated with antisense oligonucleotide. Table 26 Dose-dependent antisense inhibition of HBV RNA in HepG2.2.15 Cells using RTS3370

[000514] Some of the ISIS-oligonucleotides were also tested using the RTS3372 primer probe setup. The results are shown in Table 27. Table 27 Dose-dependent antisense inhibition of HBV RNA in HepG2.2.15 Cells using RTS3372

Example 13: Dose dependent inhibition of viral HBV RNA in HepG2.2.15 Cells by MOE gapmers

[000515] Certain gapmers from the studies described above were tested at various doses on human HepG2.2.15 cells. Cells were seeded at a density of 30,000 cells per well and transfected using LipofectAMINE 2000® reagent with concentrations of 7.8125 nM, 15.625 nM, 31.25 nM, 62.5 nM, 125.0 nM and 250.0 nM of antisense oligonucleotide, as specified in Table 28. After the treatment period of approximately 1 At 6 hours, RNA was isolated from the cells and HBV mRNA levels were measured by quantitative real-time PCR. The RTS3370 primer viral probe setup was used to measure mRNA levels. HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. Results are presented as percentage of HBV inhibition, relative to untreated control cells.

[000516] The half maximal inhibitory concentration (IC50) of each oligonucleotide is also shown in Table 28. As illustrated in Table 28, HBV mRNA levels were reduced in a dose-dependent manner in several antisense oligonucleotide treated cells. Table 28 Dose-dependent antisense inhibition of HBV RNA in HepG2.2.15 Cells using RTS3370

Example 14: Antisense inhibition of HBV viral mRNA in HepG2 cells by MOE gapmers

[000517] Antisense oligonucleotides were designed to target an HBV viral nucleic acid and were tested for their effects on HBV mRNA in vitro. The antisense oligonucleotides were tested in a series of experiments that had similar culture conditions. Results for each experiment are presented in separate tables. ISIS 146786, 509934, ISIS 509959, and ISIS 510100, from the studies described above, were also included. HepG2 cells grown at a density of 28,000 cells per well were transfected using LipofectAMINE2000® with 70 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from cells and HBV mRNA levels were measured by quantitative real-time PCR. The RTS3370 viral probe setup (forward sequence CTTGGTCATGGGCCATCAG, designated here as SEQ ID NO: 2; reverse sequence CGGCTAGGAGTTCCGCAGTA, designated here as SEQ ID NO: 3; probe sequence TGCGTGGAACCTTTTCGGCTCC, designated here as SEQ ID NO: 4) was used to measure mRNA levels. Levels were also measured using the RTS3371 primer setup (forward sequence CCAAACCTTCGGACGGAAA, designated here as SEQ ID NO: 311; reverse sequence TGAGGCCCACTCCCATAGG, designated here as SEQ ID NO: 312; probe sequence CCCATCATCCTGGGCTTTCGGAAAAT, designated here as SEQ ID NO: 313). HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. Results are presented as percentage of HBV inhibition, relative to untreated control cells. In some assays shown in Tables 32, 35, 42, 45, and 46, the potency of ISIS 146786 was measured in two wells in a single plate. In these cases, values of inhibition levels in both wells were presented.

[000518] The novel chimeric antisense oligonucleotides designated in the Tables below were designated either as gapmers 2-9-5 MOE, gapmers 2-9-6 MOE, gapmers 2-10-8 MOE, gapmers 3-94 MOE, gapmers 3-9-5 MOE, gapmers 3-10-3 MOE, gapmers 3-10-4 MOE, gapmers 3-10-7 MOE, 4-9-3 MOE gapmers, 4-9-4 MOE gapmers, 4-10-6 MOE gapmers, 5-9-2 MOE gapmers, 5-9-3 MOE gapmers, 5-10-5 MOE gapmers, 6-9-2 MOE gapmers, 6-10-4 MOE gapmers, 7-10-3 MOE gapmers, or 8-10-2 MOE gapmers. The 2-9-5 MOE gapmers are 16 nucleosides in length, where the central gap segment comprises nine 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising two and five nucleosides respectively. The 2-9-6 MOE gapmers are 17 nucleosides long, where the central gap segment comprises nine 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising two and six nucleosides respectively. The 2-10-8 MOE gapmers are 20 nucleosides long, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising two and eight nucleosides respectively. The 3-9-4 MOE gapmers are 16 nucleosides in length, where the central gap segment comprises nine 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising three and four nucleosides respectively. The 3-9-5 MOE gapmers are 17 nucleosides long, where the central gap segment comprises nine 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising three and five nucleosides respectively. The 3-10-3 MOE gapmers are 16 nucleosides in length, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising three nucleosides each. The 3-10-4 MOE gapmers are 17 nucleosides long, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising three and four nucleosides respectively. The 3-10-7 MOE gapmers are 20 nucleosides long, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising three and seven nucleosides respectively. The 4-9-3 MOE gapmers are 16 nucleosides in length, where the central gap segment comprises nine 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising four and three nucleosides respectively. 4-9-4 MOE gapmers are 17 nucleosides long, where the central gap segment comprises nine 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising four nucleosides each. The 4-10-6 MOE gapmers are 20 nucleosides long, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising four and six nucleosides respectively. The 5-9-2 MOE gapmers are 16 nucleosides in length, where the central gap segment comprises nine 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising five and two nucleosides respectively. The 5-9-3 MOE gapmers are 17 nucleosides long, where the central gap segment comprises nine 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising five and three nucleosides respectively. 5-10-5 MOE gapmers are 20 nucleosides long, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising five nucleosides each. The 6-9-2 MOE gapmers are 17 nucleosides long, where the central gap segment comprises nine 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising six and two nucleosides respectively. The 6-10-4 MOE gapmers are 20 nucleosides long, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising six and four nucleosides respectively. 7-10-3 MOE gapmers are 20 nucleosides long, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising seven and three nucleosides respectively. 8-10-2 MOE gapmers are 20 nucleosides long, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising eight and two nucleosides respectively. Each nucleoside in the 5' wing segment and each nucleoside in the 3' wing segment has an MOE sugar modification. The 'Motif' column indicates the motif with the number of nucleosides in the wings and the gap segment of each of the oligonucleotides. The internucleoside linkages across each gapmer are phosphorothioate linkages (P=S). All cytosine residues throughout each oligonucleotide are 5-methylcytosines.

[000519] "Viral target starting site" indicates the 5'-maximum nucleotide to which the gapmer is targeted in the viral gene sequence. "Viral target stop site" indicates the 3'-maximum nucleotide at which the gapmer is targeted in the viral gene sequence. The 'Motive' column indicates the gap and wing structure of each gapmer. Each gapmer listed in the Tables is a target for the viral genomic sequence, designated here as SEQ ID NO: 1 (GENBANK Accession No. U95551.1). The potency of the newly designated oligonucleotides was compared to ISIS 146786, 509934, ISIS 509959, and ISIS 510100, information of which was placed at the top of each table. Table 29 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3370 Table 30 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3370 Table 31 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3370 Table 32 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3370 Table 33 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3370 Table 34 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3370 Table 35 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3370 Table 36 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3370 Table 37 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3370 Table 38 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3371 Table 39 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3371 Table 40 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3371 Table 41 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3371 Table 42 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3371 Table 43 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3371 Table 44 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3371 Table 45 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3371 Table 46 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3371

Example 15: Antisense inhibition of HBV viral mRNA in HepG2 cells by deoxy gapmers, MOE and (S)-cEt

[000520] Antisense oligonucleotides were designed to target an HBV viral nucleic acid and were tested for their effects on HBV mRNA in vitro. The antisense oligonucleotides were tested in a series of experiments that had similar culture conditions. Results for each experiment are presented in separate tables shown below. ISIS 146786 and ISIS 509934, which were described in a previous application (Interim Application US 61/478040 filed April 21, 2011), were also included in these comparison studies. HepG2 cells grown at a density of 28,000 cells per well were transfected using LipofectAMINE2000® with 70 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from cells and HBV mRNA levels were measured by quantitative real-time PCR. The RTS3370 viral probe setup (forward sequence CTTGGTCATGGGCCATCAG, designated here as SEQ ID NO: 1; reverse sequence CGGCTAGGAGTTCCGCAGTA, designated here as SEQ ID NO: 2; probe sequence TGCGTGGAACCTTTTCGGCTCC, designated here as SEQ ID NO: 3) was used to measure mRNA levels. Levels were also measured using primer probe setup RTS3371 (forward sequence CCAAACCTTCGGACGGAAA, designated here as SEQ ID NO: 311; reverse sequence TGAGGCCCACTCCCATAGG, designated here as SEQ ID NO: 312; probe sequence CCCATCATCCTGGGCTTTCGGAAAAT, designated here as SEQ ID NO: 313). HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. Results are presented as percentage of HBV inhibition, relative to untreated control cells.

[000521] The new chimeric antisense oligonucleotides designated in the Tables below were designated as deoxy gapmers, MOE and (S)-cEt. The gapmers are 16 nucleosides in length where the nucleoside has either an MOE sugar modification, an (S)-cEt sugar modification, or a deoxy modification. The 'Chemistry' column describes the sugar modifications of each oligonucleotide. 'k' indicates a sugar modification of (S)-cEt; the number indicates the number of deoxynucleosides; and 'e' indicates a MOE modification. The internucleoside linkages across each gapmer are phosphorothioate linkages (P=S). All cytosine residues throughout each oligonucleotide are 5-methylcytosines.

[000522] "Viral target start site" indicates the 5'-maximum nucleotide to which the gapmer is targeted in the viral gene sequence. "Viral target stop site" indicates the 3'-maximum nucleotide at which the gapmer is targeted in the viral gene sequence. Each gapmer listed in the Tables is a target for the viral genomic sequence, designated herein as SEQ ID NO: 1 (GENBANK Accession No. U95551.1). The potency of the newly designed oligonucleotides was compared to ISIS 146786, 509934, ISIS 509959, and ISIS 510100. Table 47 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3370 Table 48 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3371 Table 49 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3371 Table 50 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3371 Table 51 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3370 Table 52 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3370

Example 16: Dose-dependent antisense inhibition of HBV mRNA in HepG2 cells by MOE gapmers

[000523] Antisense oligonucleotides from the study described in Example 14 exhibiting inhibition of HBV mRNA in vitro were selected and tested at various doses in HepG2 cells. Cells were seeded at a density of 28,000 cells per well and transfected using LipofectAMINE2000® with concentrations of 9.26 nM, 27.78 nM, 83.33 nM, and 250.00 nM antisense oligonucleotide, as specified in Table 53. After the approximately 16 hour treatment period, RNA was isolated from the cells and HBV mRNA levels were measured by Quantitative real-time PCR. The RTS3371 HBV primer probe setup was used to measure mRNA levels. HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. Results are presented as percentage of HBV inhibition, relative to untreated control cells.

[000524] As illustrated in Table 53, HBV mRNA levels were reduced in a dose-dependent manner in cells treated with antisense oligonucleotide. 'n/a' indicates that data for each strength is not available. Table 53 Dose-dependent antisense inhibition of human HBV in HepG2 cells

Example 17: Dose-dependent antisense inhibition of HBV mRNA in HepG2 cells by deoxy gapmers, MOE and (S)-cEt

[000525] Antisense oligonucleotides from the study described in Example 15 exhibiting inhibition of HBV mRNA in vitro were selected and tested at various doses in HepG2 cells. Cells were seeded at a density of 28,000 cells per well and transfected using LipofectAMINE2000® with concentrations of 9.26 nM, 27.78 nM, 83.33 nM, and 250.00 nM antisense oligonucleotide, as specified in Table 54. After the approximately 16 hour treatment period, RNA was isolated from cells and HBV mRNA levels were measured by Quantitative real-time PCR. The RTS3371 HBV primer probe setup was used to measure mRNA levels. HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. Results are presented as percentage of HBV inhibition, relative to untreated control cells.

[000526] As illustrated in Table 54, HBV mRNA levels were reduced in a dose-dependent manner in cells treated with antisense oligonucleotide. Table 54 Dose-dependent antisense inhibition of human HBV in HepG2 cells

Example 18: Antisense inhibition of HB V viral mRNA in HepG2 cells by deoxy gapmers, MOE and (S)-cEt

[000527] Additional antisense oligonucleotides were designed targeting an HBV viral nucleic acid and were tested for their effects on HBV mRNA in vitro. ISIS 5808 and ISIS 9591, disclosed in US5985662, as well as ISIS 146781, ISIS 146786, 524518, ISIS 552859, and ISIS 552870 were also included in these studies for comparison and are distinguished with an asterisk. HepG2 cells grown at a density of 28000 cells per well were transfected using LipofectAMINE2000® with 100 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from cells and HBV mRNA levels were measured by quantitative real-time PCR. RTS3370 and RTS3371 primer viral probe configurations were used to separately measure mRNA levels. HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. Results are presented as percentage of HBV inhibition, relative to untreated control cells.

[000528] The new chimeric antisense oligonucleotides designated in the Table below were designated as MOE gapmers or deoxy gapmers, MOE and (S)-cEt. 5-10-5 MOE gapmers are 20 nucleosides long, where the central gap segment comprises ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3' directions) by wings comprising five nucleosides each. The deoxy, MOE, and (S)-cEt gapmers are 16 nucleosides in length where the nucleoside has either a MOE sugar modification, an (S)-cEt sugar modification, or a deoxy modification. The 'Chemistry' column describes the sugar modifications of each oligonucleotide. 'k' indicates a sugar modification of (S)-cEt; the number indicates the number of deoxynucleosides; otherwise, 'd' indicates a deoxynucleoside; and 'e' indicates a MOE modification. The internucleoside linkages across each gapmer are phosphorothioate linkages (P=S). All cytosine residues throughout each oligonucleotide are 5-methylcytosines.

[000529] "Viral target starting site" indicates the 5'-maximum nucleotide to which the gapmer is targeted in the viral gene sequence. "Viral target stop site" indicates the 3'-maximum nucleotide at which the gapmer is targeted in the viral gene sequence. Each gapmer listed in Table 55 is a target for the viral genomic sequence, designated herein as SEQ ID NO: 1 (GENBANK Accession No. U95551.1). Table 55 Inhibition of HBV viral mRNA levels by chimeric antisense oligonucleotides measured with RTS3370 or RTS3371

Example 19: Efficacy of gapmers that target HBV in transgenic mice

[000530] Transgenic mice were treated with ISIS antisense oligonucleotides in a series of studies to evaluate the effectiveness of gapmers. HBV DNA and RNA levels were assessed.

Study 1

[000531] Groups of 12 mice each were injected subcutaneously twice a week for 4 weeks with 50 mg/kg of ISIS 510106, ISIS 510116, ISIS 505347, or ISIS 509934. A control group of 12 mice was injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and livers were harvested for further analysis.

DNA and RNA analysis

[000532] RNA was extracted from liver tissue for real-time PCR analysis of HBV DNA, using primer probe configurations RTS3370, RTS3371, and RTS3372. DNA levels were normalized to picogreen. HBV RNA samples were also assayed with RTS3370 and RTS3371 primer configurations after RT-PCR analysis. mRNA levels were normalized for RIBOGREEN®. Data are presented in Table 56, expressed as percentage inhibition compared to the control group. As shown in Table 56, most of the antisense oligonucleotides achieved reduction of HBV DNA and RNA under the PBS control. Results are presented as percent inhibition of HBV mRNA or DNA, relative to control. The "Chemistry" column indicates the gap-wing motive of each gapmer. Table 56 Percentage inhibition of HBV RNA and DNA in the liver of transgenic mice

Study 2

[000533] Groups of 6 mice each were injected subcutaneously twice a week for 4 weeks with 50 mg/kg of ISIS 146779, ISIS 505358, ISIS 146786, ISIS 509974, ISIS 509958, or ISIS 509959. A control group of 10 mice was injected sub cutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and livers were harvested for further analysis.

DNA and RNA analysis

[000534] RNA was extracted from liver tissue for real-time PCR analysis of HBV DNA using RTS3370 primer probe settings. DNA levels were normalized to picogreen. HBV RNA samples were also assayed with RTS3370 primer setups after RT-PCR analysis. mRNA levels were normalized for RIBOGREEN®. Data are shown in Table 57, expressed as percentage inhibition compared to the control group. As shown in Table 57, most of the antisense oligonucleotides achieved reduction of HBV DNA and RNA under the PBS control. Results are presented as percent inhibition of HBV mRNA or DNA, relative to control. The "Chemistry" column indicates the gap-wing motive of each gapmer. Table 57 Percent inhibition of HBV RNA and DNA in the liver of transgenic mice

Study 3

[000535] Groups of 6 mice each were injected subcutaneously twice a week for 4 weeks with 50 mg/kg of ISIS 509960, ISIS 505329, ISIS 146786, ISIS 505339, or ISIS 509927. Another group of 6 mice was administered Entecavir, an oral antiviral drug used to treat Hepatitis B infection, at 1 mg/kg daily for two weeks. A control group of 10 mice was injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and livers were harvested for further analysis.

DNA and RNA analysis

[000536] RNA was extracted from liver tissue for real-time PCR analysis of HBV DNA using RTS3371 primer probe settings. DNA levels were normalized to picogreen. HBV RNA samples were also assayed with RTS3371 primer setups after RT-PCR analysis. mRNA levels were normalized for RIBOGREEN®. Data are presented in Table 58, expressed as percentage inhibition compared to the control group. As shown in Table 58, most of the antisense oligonucleotides achieved reduction of HBV DNA and RNA under the PBS control. Results are presented as percent inhibition of HBV mRNA or DNA, relative to control. The "Chemistry" column indicates the gap-wing motive of each gapmer. Table 58 Percentage inhibition of HBV RNA and DNA in the liver of transgenic mice

Study 4

[000537] Groups of 6 mice each were injected subcutaneously twice a week for 4 weeks with 25 mg/kg of ISIS 146786, ISIS 552176, and ISIS 552073. One group of 10 mice was injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and organs and plasma were harvested for further analysis.

DNA and RNA analysis

[000538] RNA was extracted from liver tissue for real-time PCR analysis of HBV DNA, using RTS3371 primer probe setup. DNA levels were normalized to picogreen. HBV RNA samples were also assayed with the RTS3371 primer configuration after RT-PCR analysis. mRNA levels were normalized for RIBOGREEN®. The data are shown in Table 59. As shown in Table 59, the antisense oligonucleotides achieved reduction of HBV DNA and RNA over the PBS control. Results are presented as percent inhibition of HBV mRNA or DNA, relative to control. The "Chemistry" column indicates the gap-wing motive of each gapmer. Table 59 Percentage inhibition of HBV RNA and DNA in transgenic mice

Liver Function

[000539] To assess the effect of ISIS oligonucleotides on liver function, plasma ALT concentrations were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY) ( Nyblom, H. et al., Alcohol & Alcoholism 39: 336-339, 2004; Tietz NW (Ed): Clinical Guide to Laboratory Tests, 3rd ed. WB Saunders , Philadelphia, PA, 1995). The results are shown in Table 60 expressed in IU/L. Both ISIS oligonucleotides were considered tolerable in mice, as demonstrated by their liver transaminase profile. Table 60 ALT levels (IU/L) of transgenic mice

Study 5

[000540] Groups of 6 mice each were injected subcutaneously twice a week for 4 weeks with 25 mg/kg of ISIS 146786, ISIS 552056, ISIS 552088, and ISIS 552309. One group of 10 mice was injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and organs and plasma were harvested for further analysis.

DNA and RNA analysis

[000541] RNA was extracted from liver tissue for real-time PCR analysis of HBV DNA, using RTS3371 primer probe setup. DNA levels were normalized to picogreen. HBV RNA samples were also assayed with the RTS3371 primer configuration after RT-PCR analysis. mRNA levels were normalized for RIBOGREEN®. As shown in Table 61, the antisense oligonucleotides achieved reduction of HBV DNA and RNA over the PBS control. Results are presented as percent inhibition of HBV mRNA or DNA, relative to control. The "Chemistry" column indicates the gap-wing motive of each gapmer. Table 61 Percentage inhibition of HBV DNA and RNA in transgenic mice

Study 6

[000542] Groups of 6 mice each were injected subcutaneously twice a week for 4 weeks with 25 mg/kg of ISIS 146786, ISIS 505330, ISIS 509932, ISIS 552032, ISIS 552057, ISIS 552075, ISIS 552092, and ISIS 5522 55. A group of 10 mice were injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and organs and plasma were harvested for further analysis.

DNA and RNA analysis

[000543] RNA was extracted from liver tissue for real-time PCR analysis of HBV DNA, using RTS3371 primer probe setup. DNA levels were normalized to picogreen. HBV RNA samples were also assayed with the RTS3371 primer configuration after RT-PCR analysis. mRNA levels were normalized for RIBOGREEN®. As shown in Table 62, the antisense oligonucleotides achieved reduction of HBV DNA and RNA over the PBS control. Results are presented as percent inhibition of HBV mRNA or DNA, relative to control. The "Chemistry" column indicates the gap-wing motive of each gapmer. Table 62 Percentage inhibition of HBV DNA and RNA in transgenic mice

Study 7

[000544] Groups of 6 mice each were injected subcutaneously twice a week for 4 weeks with 20 mg/kg of ISIS 552859, ISIS 577121, ISIS 577122, ISIS 577123, ISIS 577132, ISIS 577133, and ISIS 577134. soxy, MOE and (S)-cEt. A group of 10 mice were injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and organs and plasma were harvested for further analysis.

DNA and RNA analysis

[000545] RNA was extracted from liver tissue for real-time PCR analysis of HBV DNA, using RTS3371 primer probe setup. DNA levels were normalized to picogreen. HBV RNA samples were also assayed with the RTS3371 primer configuration after RT-PCR analysis. mRNA levels were normalized for RIBOGREEN®. As shown in Table 63, the antisense oligonucleotides achieved reduction of HBV DNA and RNA over the PBS control. Results are presented as percent inhibition of HBV mRNA or DNA, relative to control. The 'Chemistry' column describes the sugar modifications of each oligonucleotide. 'k' indicates a sugar modification of (S)-cEt; the number indicates the number of deoxynucleosides; otherwise, 'd' indicates a deoxynucleoside; and 'e' indicates a MOE modification. Table 63 Percentage inhibition of HBV DNA and RNA in transgenic mice

Study 8

[000546] A group of 6 mice was injected subcutaneously twice a week for 4 weeks with 25 mg/kg of ISIS 146786, the gapmer 5-10-5 MOE. Groups of 6 mice each were injected subcutaneously twice a week for 4 weeks with 10 mg/kg of ISIS 552803, ISIS 552903, ISIS 552817, ISIS 552822, and ISIS 552907. These gapmers all had deoxy, MOE, and (S)-cEt chemistry. A group of 10 mice were injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and organs and plasma were harvested for further analysis.

DNA and RNA analysis

[000547] RNA was extracted from liver tissue for real-time PCR analysis of HBV DNA, using RTS3371 primer probe setup. DNA levels were normalized to picogreen. HBV RNA samples were also assayed with the RTS3371 primer configuration after RT-PCR analysis. mRNA levels were normalized for RIBOGREEN®. The data are shown in Table 64. As shown in Table 64, the antisense oligonucleotides achieved reduction of HBV DNA and RNA over the PBS control. The 'Chemistry' column describes the sugar modifications of each oligonucleotide. 'k' indicates a sugar modification of (S)-cEt; the number indicates the number of deoxynucleosides; otherwise, 'd' indicates a deoxynucleoside; and 'e' indicates a MOE modification; in the case of MOE gapmers, the "Chemistry" column defines the gap-wing structure. Table 64 Percentage inhibition of HBV RNA and DNA in transgenic mice

Study 9

[000548] A group of 6 mice were injected subcutaneously twice a week for 4 weeks with 25 mg/kg of ISIS 146786. Groups of 6 mice each were injected subcutaneously twice a week for 4 weeks with 10 mg/kg of ISIS 552853, ISIS 552854, ISIS 552932, and ISIS 5 52938. A group of 10 mice were injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and organs and plasma were harvested for further analysis.

DNA and RNA analysis

[000549] RNA was extracted from liver tissue for real-time PCR analysis of HBV DNA, using RTS3371 primer probe setup. DNA levels were normalized to picogreen. HBV RNA samples were also assayed with the RTS3371 primer configuration after RT-PCR analysis. mRNA levels were normalized for RIBOGREEN®. As shown in Table 65, the antisense oligonucleotides achieved reduction of HBV DNA and RNA over the PBS control. Results are presented as percent inhibition of HBV mRNA or DNA, relative to control. The 'Chemistry' column describes the sugar modifications of each oligonucleotide. 'k' indicates a sugar modification of (S)-cEt; the number indicates the number of deoxynucleosides; otherwise, 'd' indicates a deoxynucleoside; and 'e' indicates a MOE modification; in the case of MOE gapmers, the "Chemistry" column defines the gap-wing structure. Table 65 Percentage inhibition of HBV DNA and RNA in transgenic mice

Study 10

[000550] A group of 6 mice were injected subcutaneously twice a week for 4 weeks with 25 mg/kg of ISIS 146786. Groups of 6 mice each were injected subcutaneously twice a week for 4 weeks with 10 mg/kg of ISIS 552922, ISIS 552923, ISIS 552942, ISIS 55 2872, ISIS 552925, ISIS 552937, and ISIS 552939. A group of 10 mice were injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and organs and plasma were harvested for further analysis.

DNA and RNA analysis

[000551] RNA was extracted from liver tissue for real-time PCR analysis of HBV DNA, using RTS3371 primer probe setup. DNA levels were normalized to picogreen. HBV RNA samples were also assayed with the RTS3371 primer configuration after RT-PCR analysis. mRNA levels were normalized for RIBOGREEN®. As shown in Table 66, the antisense oligonucleotides achieved reduction of HBV DNA and RNA over the PBS control. Results are presented as percent inhibition of HBV mRNA or DNA, relative to control. The 'Chemistry' column describes the sugar modifications of each oligonucleotide. 'k' indicates a sugar modification of (S)-cEt; the number indicates the number of deoxynucleosides; otherwise, 'd' indicates a deoxynucleoside; and 'e' indicates a MOE modification; in the case of MOE gapmers, the "Chemistry" column defines the gap-wing structure. Table 66

Example 20: Efficacy of gapmers that target HBV in transgenic mice

[000552] Mice carrying the HBV gene fragment (Guidotti, L.G. et al., J. Virol. 1995, 69, 6158-6169) were used. Mice were treated with ISIS antisense oligonucleotides selected from the studies described above and evaluated for their effectiveness in this model. HBV DNA, RNA, and antigen levels were assessed.

[000553] Groups of 10 mice each were injected subcutaneously twice a week during the first with 50 mg/kg and subsequently twice a week for the next 3 weeks with 25 mg/kg of ISIS 146786 or ISIS 510100. Control groups of 10 mice each were treated in a similar manner with ISIS 141923 (CCTTCCCTGAA GGTTCCCTCC, SEQ ID NO: 320; gapmer 5-10-5 MOE with no known murine target) or ISIS 459024 (CGGTCCTTGGAGGATGC, SEQ ID NO: 1351; gapmer 3-10-4 MOE with no known murine target). Mice were euthanized 48 hours after the last dose, and organs and serum were harvested for further analysis.

DNA and RNA analysis

[000554] RNA was extracted from liver tissue for real-time PCR analysis of HBV DNA, using primer probe settings RTS3370, RTS3371, or RTS3372 (direct sequence ATCCTATCAACACTTCCGGAAACT, designated SEQ ID NO: 314; reverse sequence CGACGCGGCGATTGAG, designated SEQ ID NO: 315; probe sequence AAGAACTCCCTCGCC TCGCAGACG, designated SEQ ID NO: 316). DNA levels were normalized to picogreen. HBV RNA samples were also assayed with RTS3370 and RTS3371 primer configurations after RT-PCR analysis. mRNA levels were normalized for RIBOGREEN®. Data are presented in Table 67. Serum DNA samples were analyzed after the study period. The data are presented in Table 68, they were expressed in relation to the levels measured in the control group. As shown in Tables 67 and 68, the antisense oligonucleotides achieved reduction of HBV DNA and RNA over the PBS control. Results are presented as percent inhibition of HBV mRNA or DNA, relative to control. The "Chemistry" column defines the gap-wing structure of each gapmer. Table 67 Percentage inhibition of HBV RNA and DNA in the liver of transgenic mice Table 68 Percentage of inhibition of HBV DNA in serum from transgenic mice

HBV antigen analysis

[000555] The HBV antigens in the supernatants were detected using the ELISA technique. HBs antigen (HBsAg) levels were detected by ELISA from Abazyme LLC, MA. As shown in Table 57, treatment with oligonucleotides ISIS 146786 or 510100 caused a reduction in HBsAg levels. HBe antigen (HBeAg) levels were detected by ELISA from International Immuno-diagnostics, CA. As shown in Table 69, treatment with oligonucleotides ISIS 146786 or 510100 also caused a reduction in HBeAg levels. Table 69 HBV antigen levels (PEI U/mL) in transgenic mice

Example 21: Antisense inhibition of HBV viral mRNA in HepG2 cells by deoxy gapmers, MOE and (S)-cEt

[000556] Additional antisense oligonucleotides were designed targeting an HBV viral nucleic acid and were tested for their effects on HBV mRNA in vitro. ISIS 146786, ISIS 505358, ISIS 509932, and ISIS 510100, disclosed in the U.S. Provisional Application No. 61/478,040 filed on April 21, 2011; ISIS 552859 disclosed in the U.S. No. 61/596,692 filed on February 8, 2012; ISIS 577121, ISIS 577122, ISIS 577123, ISIS 577132, ISIS 577133, and ISIS 577134, disclosed in the study described above, were also included in the trial. HepG2 cells grown at a density of 28,000 cells per well were transfected using Cytofectin with 9.375 nM, 18.75 nM, 37.50 nM, 75.00 nM, 150.00 nM, or 300.00 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from cells and HBV mRNA levels were measured by quantitative real-time PCR. The RTS3371 primer viral probe setup was used to measure mRNA levels. HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. Results are presented as percentage of HBV inhibition, relative to untreated control cells.

[000557] The novel chimeric antisense oligonucleotides designated in the Tables below were designated as deoxy gapmers, MOE and (S)-cEt. The deoxy, MOE, and (S)-cEt gapmers are 16, 17, or 18 nucleosides in length where the nucleosides have either a MOE sugar modification, an (S)-cEt sugar modification, or a deoxy modification. The 'Chemistry' column describes the sugar modifications of each oligonucleotide. 'k' indicates a sugar modification of (S)-cEt; the number indicates the number of deoxynucleosides; otherwise, 'd' indicates a deoxynucleoside; and 'e' indicates a MOE modification. The internucleoside linkages across each gapmer are phosphorothioate linkages (P=S). All cytosine residues throughout each oligonucleotide are 5-methylcytosines.

[000558] "Viral target starting site" indicates the 5'-maximum nucleotide to which the gapmer is targeted in the viral gene sequence. "Viral target stop site" indicates the 3'-maximum nucleotide at which the gapmer is targeted in the viral gene sequence. Each gapmer listed in Table 70 is a target for the viral genomic sequence, designated herein as SEQ ID NO: 1 (GENBANK Accession No. U95551.1). Table 70 Chimeric Antisense Oligonucleotides Targeting SEQ ID NO: 1 Table 71 Dose-dependent inhibition of HBV mRNA levels by chimeric antisense oligonucleotides Table 72 Dose-dependent inhibition of HBV mRNA levels by chimeric antisense oligonucleotides

Example 22: Analysis of the potency of uniform deoxyoligonucleotides in inhibiting HBV mRNA in HepG2 cells

[000559] Additional antisense oligonucleotides were tested for their effects on HBV mRNA in vitro. ISIS 5808 and ISIS 9591, disclosed in US5985662 were also included in the test. ISIS 146786 was included in the test as the benchmark. HepG2 cells grown at a density of 28,000 cells per well were transfected using LipofectAMINE2000® with 18.75 nM, 37.50 nM, 75.00 nM, 150.00 nM, or 300.00 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from cells and HBV mRNA levels were measured by quantitative real-time PCR. The RTS3371 primer viral probe setup was used to measure mRNA and DNA levels. HBV mRNA levels were adjusted for total RNA content as measured by RIBOGREEN®. S antigen and E antigen levels were also measured by ELISA. Results are presented as percent inhibition versus untreated control cells.

[000560] The tested antisense oligonucleotides, ISIS 582699, ISIS 582700, and ISIS 582701, were named according to the Sequences and chemistries disclosed in Korba and Gerin, Antiviral Research, 1995, Vol. 28, 225-242; the corresponding names for the oligonucleotides in the reference are S1, C1, and L2c, respectively. The antisense oligonucleotides in the Tables below have been designated as uniform deoxy oligonucleotides, 16 or 21 nucleosides in length where the nucleosides have deoxy modifications. "Viral target starting site" indicates the 5'-maximum nucleotide to which the oligonucleotide is targeted in the viral gene sequence. "Viral target stop site" indicates the 3'-maximum nucleotide at which the oligonucleotide is targeted in the viral gene sequence. Each oligonucleotide listed in Table 73 targets the viral genomic sequence, designated herein as SEQ ID NO: 1 (GENBANK Accession No. U95551.1). The results indicate that deoxy oligonucleotides had negligible effect on HBV mRNA and HBV DNA antigen expression levels. Table 73 Uniform Deoxy Oligonucleotides Targeting SEQ ID NO: 1 Table 74 Dose-dependent inhibition of HBV mRNA levels after treatment with oligonucleotides Table 75 Dose-dependent inhibition of HBV DNA levels in HepG2 cells after treatment with oligonucleotides Table 76 HBV S antigen levels after treatment with oligonucleotides (arbitrary units) Table 77 HBV E antigen levels after treatment with oligonucleotides (arbitrary units)

Claims (14)

1. Compound, characterized in that it comprises a modified single-stranded oligonucleotide consisting of 20 linked nucleosides and having a nucleobase sequence consisting of any of the nucleobase sequences of SEQ ID NOs: 226, 224, and 807, wherein at least one modified single-stranded oligonucleotide nucleoside comprises a modified sugar and the modified single-stranded oligonucleotide is 100% complementary to SEQ ID NO:1, being that the modified sugar comprises a 2'-O-methoxyethyl group or a bicyclic sugar. 2. Compound according to claim 1, characterized in that the modified single-stranded oligonucleotide has the nucleobase sequence of SEQ ID NO: 226. 3. Compound according to claim 1 or 2, characterized in that the modified single-stranded oligonucleotide comprises at least one phosphorothioate internucleoside bond. 4. Compound according to claim 3, characterized in that each internucleoside bond is a phosphorothioate internucleoside bond. 5. Compound according to any one of claims 1 to 4, characterized in that the at least one modified sugar comprises a 2'-O-methoxyethyl group. 6. Compound according to any one of claims 1 to 4, characterized in that the at least one modified sugar comprises a bicyclic sugar. 7. Compound according to any one of claims 1 to 6, characterized in that the at least one nucleoside comprises a modified nucleobase. 8. Compound according to claim 7, characterized in that the modified nucleobase is a 5-methylcytosine. 9. Compound according to any one of claims 1 to 8, characterized in that the modified oligonucleotide comprises: a gap segment consisting of linked deoxynucleosides; a 5' wing segment consisting of linked nucleosides; and a 3' wing segment consisting of linked nucleosides; wherein the gap segment is positioned between the 5' wing segment and the 3' wing segment, and wherein each nucleoside of each wing segment comprises a modified sugar. 10. Compound according to claim 1, characterized in that the modified single-stranded oligonucleotide consists of 20 linked nucleosides having the nucleobase sequence of SEQ ID NO: 226 and comprises: a gap segment consisting of 10 linked deoxynucleosides; a 5' wing segment consisting of 5 linked nucleosides; and a 3' wing segment consisting of 5 linked nucleosides; wherein the gap segment is positioned between the 5' wing segment and the 3' wing segment, wherein each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar, wherein each internucleoside bond is a phosphorothioate bond, and wherein each cytosine is a 5-methylcytosine. 11. Compound according to any one of claims 1 to 10, characterized in that the compound consists of a modified single-stranded oligonucleotide. 12. Pharmaceutical composition, characterized in that it comprises the compound, as defined in any one of claims 1 to 11, or a pharmaceutically acceptable derivative thereof, and at least one of a pharmaceutically acceptable vehicle or diluent. 13. Use of the compound, as defined in any one of claims 1 to 11, or of a composition, as defined in claim 12, characterized in that it is for the preparation of a medicament for the treatment of a disease, a disorder or a condition related to HBV in an individual, in which the disease, disorder or condition is chronic HBV infection, liver cancer, and diffuse hepatocellular inflammatory disease. 14. Use of the compound, as defined in any one of claims 1 to 11, or of a composition, as defined in claim 12, characterized in that it is for the preparation of a medicine for the treatment of chronic HBV infection.