FITZGERALD MEGAN ELIZABETH (US)
MONTERO SAUL MARTINEZ (US)
BHATTACHARYA ANEERBAN (US)
WO2017157899A1 | 2017-09-21 | |||
WO2017100587A1 | 2017-06-15 | |||
WO2019004939A1 | 2019-01-03 | |||
WO2018065589A1 | 2018-04-12 |
SCOLES, D. ET AL.: "Antisense oligonucleotides", NEUROLOGY:GENETICS, vol. 5, April 2019 (2019-04-01), XP055712472, DOI: 10.1212/NXG.0000000000000323
HUANG, Y.: "Preclinical and Clinical advances of GalNAc-Decorated Nucleic Acid Therapeutics", MOLECULAR THERAPY:NUCLEIC ACIDS, vol. 6, March 2017 (2017-03-01), pages 116 - 132, XP055485332, DOI: 10.1016/j.omtn.2016.12.003
WHAT IS CLAIMED IS: 1. An antisense oligonucleotide (ASO) that targets human CD274 mRNA, wherein the ASO comprises 14 to 20 nucleotides selected from the group consisting of unmodified nucleotides and modified nucleosides, wherein each of the modified nucleosides (a) contains a modified sugar, (b) contains a modified nucleobase or is abasic, or (c) both contains a modified sugar and contains a modified nucleobase or is abasic; wherein each linkage between the nucleosides is a phosphorothioate, phosphodiester, phosphoramidate, thiophosphoramidate, methylphosphate, methylphosphonate, phosphonoacetate, boranophosphate, or any combination thereof; and wherein the ASO is at least 85% complementary to a fragment of human CD274 mRNA. 2. The ASO of claim 1, wherein the ASO comprises: (a) zero nucleotide mismatches if the ASO is 14 or 15 nucleotides in length; (b) up to one nucleotide mismatch if the ASO is m nucleotides in length at position 1, 2, 3, m-2, m-1, or m, wherein m is 16 or 17; (c) up to two nucleotide mismatches if the ASO is m nucleotides in length at position 1, 2, 3, 4, m-3, m-2, m-1, or m, wherein m is 18 or 19; or (d) up to two nucleotide mismatches if the ASO is 20 nucleotides in length at position 1, 2, 3, 4, 5, 16, 17, 18, 19, or 20. 3. The ASO of claim 1 or 2, wherein the ASO has a sequence as set forth in any one of SEQ ID NOs: 2-301. 4. The ASO of claim 1 or 2, wherein the ASO has 14 nt. 5. The ASO of claim 1 or 2, wherein the ASO has 15 nt. 6. The ASO of claim 1 or 2, wherein the ASO has 16 nt. 7. The ASO of claim 1 or 2, wherein the ASO has 17 nt. 8. The ASO of claim 1 or 2, wherein the ASO has 18 nt. 9. The ASO of claim 1 or 2, wherein the ASO has 19 nt. 10. The ASO of claim 1 or 2, wherein the ASO has 20 nt. 11. The ASO of any one of claims 1-10, wherein the modified sugar is selected from the group consisting of 2’-OMe, 2’-F, 2’-MOE, 2’-araF, 2’-araOH, 2’-OEt, 2’-O-alkyl, LNA, scpBNA, AmNA, cEt, and ENA. 12. The ASO of any one of claims 1-11, wherein the modified nucleobase is selected from the group consisting of 5-OH-C, 2S-T, 8-NH2-A, 8-NH2-G, and 5-methyl-C. 13. The ASO of any one of claims 1-12, further comprising a targeting or lipophilic moiety. 14. The ASO of claim 13, wherein the targeting or lipophilic moiety is conjugated to the ASO at the 5’ end, 3’ end, or both. 15. The ASO of any one of claims 13-14, wherein the targeting or lipophilic moiety is GalNAc, folic acid, cholesterol, tocopherol, palmitate, or a long chain fatty acid. 16. The ASO according to any one of claims 1-15, wherein the ASO is a gapmer, mixmer, or blockmer. 17. The ASO of any one of claim 1-16, wherein the modified nucleosides are selected from the group consisting of: , , , , wherein R1 is hydrogen or C1-7 alkyl. 18. The ASO of claim 17, wherein the Base is selected from the group consisting of adenine, guanine, cytosine, thymine, uracil, pseudouracil, 2-thio-uracil, dihydrouracil, 5- bromo-uracil, 5-iodo-uracil, 5’-methyl-cytosine, 7-deazapurine, 2,6-diaminopurine, inosine, phenoxazine . 19. The ASO of claim 18, wherein the modified nucleobase is selected from the group consisting of pseudouracil, 2-thio-uracil, dihydrouracil, 5-bromo-uracil, 5-iodo-uracil, 5-methyl-cytosine, 7-deazapurine, 2,6-diaminopurine, inosine, phenoxazine and . 20. A pharmaceutical composition comprising an effective amount of the ASO according to any one of claims 1-19 and a pharmaceutically acceptable carrier, diluent, excipient, or combination thereof. 21. An ASO of any one of claims 1-19 for use in treating hepatitis B. 22. An ASO of any one of claims 1-19 for use in treating hepatocellular carcinoma (HCC). 23. The ASO of any one of claims 21-22, wherein the ASO is used in combination with surgery, radiation therapy, chemotherapy, targeted therapy, immunotherapy, hormonal therapy, or antiviral therapy. 24. The ASO of claim 23, wherein the ASO comprises an ASO against PD-L1 and an ASO or an siRNA against hepatitis B virus (HBV). 25. A method for treating hepatitis B in a subject comprising administering to the subject in need thereof an effective amount of an ASO of any one of claims 1-19 or administering to the subject in need thereof an effective amount of a pharmaceutical composition of claim 20. 26. A method for treating hepatocellular carcinoma (HCC) in a subject comprising administering to the subject in need thereof an effective amount of an ASO of any one of claims 1-19 or administering to the subject in need thereof an effective amount of a pharmaceutical composition of claim 20. 27. The method of any one of claims 25-26, further comprising administering surgery, radiation therapy, chemotherapy, targeted therapy, immunotherapy, hormonal therapy, or antiviral therapy. |
wherein R is OH or SH, and wherein n is any integer. In some embodiments, the deoxycytosine nucleotide shown in this structure linking the ASO to the GalNAc moiety is optional, and can be omitted. In some embodiments, n ranges from 0 to 10, such as 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. For example, for GalNAc4, n=1; and for GalNAc6, n=2. However, it is to be understood that n may equal any integer and may be selected based on the desired characteristic of the targeting moiety. [0069] As used herein, the term “gapmer” refers to an ASO that comprises both deoxyribose (DNA) and ribose (RNA)-based nucleotides wherein the DNA nucleotides are grouped together in the middle of the ASO sequence and flanked by the RNA nucleotides on both 5’ and 3’ ends. The DNA nucleotides are involved in RNase H-mediated cleavage of the target mRNA. Each of the flanking RNA nucleotides can be unmodified nucleotides or modified nucleotides to enhance ASO properties such as stability, resistance to nuclease degradation, base pairing efficiency, solubility, conformation/flexibility of the oligonucleotide, or melting temperature. The DNA nucleotides can also be modified nucleotides if the modifications do not affect base pairing to the target mRNA or RNase H activity to the desired threshold. The gapmer can also comprise base pair mismatches to the target sequence. In some embodiments, the base pair mismatches are in the outer RNA nucleotides, while the center DNA or RNA nucleotides of the gapmer are conserved. [0070] As used herein, the term “mixmer” refers to an ASO that comprises both unmodified and modified DNA or RNA nucleotides wherein the unmodified nucleotides and modified nucleotides are distributed throughout the ASO sequence. In some embodiments, the unmodified nucleotides and modified nucleotides are alternating in the ASO sequence. In some embodiments, two, three, four, five, or six unmodified nucleotides, or two, three, four, five, or six modified nucleotides are grouped together in a sequence that otherwise contains alternating unmodified and modified nucleotides. Groups of contiguous unmodified nucleotides or contiguous modified nucleotides can be spaced across the ASO at regular intervals. [0071] As used herein, the term “blockmer” refers to an ASO that comprises both unmodified and modified DNA or RNA nucleotides wherein the unmodified nucleotides are grouped together, and the modified nucleotides are grouped together. In some embodiments, the blockmer can comprise 1, 2, 3, 4, 5, 6, 7, or 8 unmodified nucleotides on the 5’ end of the ASO. In some embodiments, the blockmer can comprise 1, 2, 3, 4, 5, 6, 7, or 8 modified nucleotides on the 5’ end of the ASO. In some embodiments, the blockmer can comprise 1, 2, 3, 4, 5, 6, 7, or 8 unmodified nucleotides on the 3’ end of the ASO. In some embodiments, the blockmer can comprise 1, 2, 3, 4, 5, 6, 7, or 8 modified nucleotides on the 3’ end of the ASO. [0072] As used herein, the term “Xmer” refers to an oligonucleotide or nucleic acid polymer that is “X” nucleotides long. For example, a 14mer is an oligonucleotide or nucleic acid polymer that is 14 nucleotides long, and a 20mer is an oligonucleotide or nucleic acid polymer that is 20 nucleotides long. In some embodiments, the “X” refers to the total number of nucleotides. In other embodiments, the “X” refers to the number of nucleotides involved in binding to the target, while the oligonucleotide or nucleic acid polymer may have additional nucleotides or components that are not involved in binding to the target. [0073] In some embodiments, at least one ASO is used to treat liver disease. In some embodiments, the liver disease includes but is not limited to liver cancer, hepatocellular carcinoma (HCC), cholangiocarcinoma, hepatitis, hepatitis A, hepatitis B, hepatitis C, hepatitis D, or any combination thereof. In some embodiments, the at least one ASO is used to silence expression of a gene involved in a liver disease. In some embodiments, the gene is CD274. In some embodiments, the at least one ASO results in at least 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% reduction in the disease or symptoms thereof. [0074] The term “isolated” as used herein refers to material that is substantially or essentially free from components that normally accompany it in its native state. For example, an “isolated cell,” as used herein, includes a cell that has been purified from the milieu or organisms in its naturally occurring state, a cell that has been removed from a subject or from a culture, for example, it is not significantly associated with in vivo or in vitro substances. [0075] As used herein, the abbreviations for any protective groups and other compounds are used, unless indicated otherwise, in accord with their common usage. [0076] It is to be understood that where compounds disclosed herein have unfilled valencies, then the valencies are to be filled with hydrogen or isotopes thereof, e.g., hydrogen-1 (protium) and hydrogen-2 (deuterium). [0077] It is understood that the compounds described herein can be labeled isotopically. Substitution with isotopes such as deuterium may afford certain therapeutic advantages resulting from greater metabolic stability, such as, for example, increased in vivo half-life or reduced dosage requirements. Each chemical element as represented in a compound structure may include any isotope of said element. For example, in a compound structure a hydrogen atom may be explicitly disclosed or understood to be present in the compound. At any position of the compound that a hydrogen atom may be present, the hydrogen atom can be any isotope of hydrogen, including but not limited to hydrogen-1 (protium) and hydrogen-2 (deuterium). Thus, reference herein to a compound encompasses all potential isotopic forms unless the context clearly dictates otherwise. [0078] Where a range of values is provided, it is understood that the upper and lower limit, and each intervening value between the upper and lower limit of the range is encompassed within the embodiments. Oligonucleotide Synthesis [0079] Each of 2’-OMe, 2’-MOE, and LNA phosphoramidite monomers were procured from commercially-available sources. All the monomers were dried in vacuum desiccator with desiccants (P2O5, RT 24h). Universal solid supports (CPG) attached were obtained from ChemGenes. The chemicals and solvents for synthesis workflow were purchased from VWR/Sigma commercially-available sources and used without any purification or treatment. Solvent (Acetonitrile) and solutions (amidite and activator) were stored over molecular sieves during synthesis. [0080] The control and target oligonucleotide sequences were synthesized on an Expedite 8909 synthesizer using the standard cycle written by the manufacturer with modifications as needed to wait steps and coupling steps. The solid support was controlled pore glass and the monomers contained standard protecting groups. Each chimeric oligonucleotide was individually synthesized using commercially available 5'-O-(4,4'- dimethoxytrityl)-3'-O-(2-cyanoethyl-N, N-diisopropyl) DNA, 2’-OMe, 2’-MOE and or LNA phosphoramidite monomers of 6-N-benzoyladenosine (A Bz ), 4-N-acetylcytidine (C Ac ), 2-N- isobutyrylguanosine (G iBu ), and Uridine (U) or Thymidine (T), according to standard solid phase phosphoramidite synthesis protocols. The 2’-O-Me-2,6, diaminopurine phosphoramidite was purchased from Glen Research. The phosphoramidites were prepared as 0.1 M solutions in anhydrous acetonitrile. 5-Ethylthiotetrazole was used as activator, 3% Dichloroacetic acid in dichloromethane was used to detritylate, acetic anhydride in THF and 16% N-methylimidazole in THF were used to cap, and DDTT ((dimethylamino-methylidene) amino)-3H-1,2,4-dithiazaoline-3-thione was used as the sulfur-transfer agent for the synthesis of oligoribonucleotide phosphorothioates. An extended coupling of 0.1M solution of phosphoramidite in CH 3 CN in the presence of 5-(ethylthio)-1H-tetrazole activator to a solid bound oligonucleotide followed by extended capping, oxidation and deprotection to afford the modified oligonucleotides. The stepwise coupling efficiency of all modified phosphoramidites was more than 98.5%. [0081] Deprotection and cleavage from the solid support was achieved with mixture of ammonia methylamine (1:1, AMA) for 15 min at 65°C, when the universal linker was used, the deprotection was left for 90 min at 65°C or solid supports were heated with aqueous ammonia (28%) solution at 55°C for 8 h to deprotect the base labile protecting groups. After filtering to remove the solid support, the deprotection solution was removed under vacuum in a GeneVac centrifugal evaporator. Tables 1-3 depicts exemplary structures of 2’-OMe, 2’-MOE, and LNA phosphoramidite monomers Table 1: 2’-OMe Phosphoramidite Monomers Table 2: 2’-MOE Phosphoramidite Monomers Table 3: 2’-LNA Phosphoramidite Monomers
[0082] The AmNA and Scp-BNA phosphoramidite monomers of 6-N- benzoyladenosine (A Bz ), 4-N-acetylcytidine (C Ac ), 2-N-isobutyrylguanosine (G iBu ), and Thymidine (T) received from LUXNA Technologies. All the monomers were dried in a vacuum desiccator with desiccants (P 2 O 5 , at room temperature for 24 hours). For the AmNA- PS-DNA-PS and scp-BNA-PS-DNA-PS modifications, the synthesis was carried out on a 1 μM scale in a 3’ to 5’ direction with the phosphoramidite monomers diluted to a concentration of 0.12 M in anhydrous CH3CN in the presence of 0.3 M 5-(benzylthio)-1H- tetrazole activator (coupling time 16-20 min) to a solid bound oligonucleotide followed by modified capping, oxidation and deprotection to afford the modified oligonucleotides. The stepwise coupling efficiency of all modified phosphoramidites was more than 97%. The DDTT (dimethylamino-methylidene) amino)-3H-1, 2, 4-dithiazaoline-3-thione was used as the sulfur-transfer agent for the synthesis of the oligoribonucleotide phosphorothioates. Oligonucleotide-bearing solid supports were washed with 20% DEA solution in acetonitrile for 15 min then the column was washed thoroughly with AcCN. The support was heated at 65 o C with Diisopropylamine:water:Methanol (1:1:2) for 5 h in heat block to cleave from the support and deprotect the base labile protecting groups. Tables 4 and 5 depicts exemplary structures of the AmNA and Scp-BNA phosphoramidite monomers. Table 4: am-NCH 3 Phosphoramidite Monomers Table 5: Scp-BNA Phosphoramidite Monomers [0083] The cholesterol, tocopherol phosphoramidite, and solid supports were received from ChemGenes. The cholesterol and Tocopherol conjugated oligonucleotides were obtained by starting solid phase synthesis on cholesterol and Tocopherol supports attached on TEG linker for 3’-conjugation while final coupling of the phosphoramidite provided the 5’-conjugated oligonucleotides. Quantitation of Crude Oligomer or Raw Analysis [0084] Samples were dissolved in deionized water (1.0mL) and quantified as follows: Blanking was first performed with water alone (1.0 mL), then 20 μL of sample and 980 μL of water were mixed well in a microfuge tube, transferred to cuvette and absorbance reading obtained at 260 nm. The crude material was dried and stored at -20°C. Crude HPLC/LC-MS analysis [0085] The 0.1 OD of the crude samples were used for crude MS analysis. After confirming the crude LC-MS data, the purification step was performed. HPLC Purification [0086] The Phosphodiester (PO), Phosphorothioate (PS) and chimeric modified oligonucleotides were purified by anion-exchange HPLC. The buffers were 20 mM sodium phosphate in 10 % CH 3 CN, pH 8.5 (buffer A) and 20 mM sodium phosphate in 10% CH 3 CN, 1.8 M NaBr, pH 8.5 (buffer B). Fractions containing full-length oligonucleotides were pooled, desalted and lyophilized. [0087] The conjugated oligonucleotides were purified by an in-house packed RPC-Source15 reverse-phase column. The buffers were 20 mM sodium acetate in 10% CH3CN, (buffer A) and CH3CN (buffer B). Fractions containing full-length oligonucleotides were pooled, desalted and lyophilized. Desalting of Purified Oligomer [0088] The purified dry oligomer was then desalted using Sephadex G-25 M (Amersham Biosciences). The cartridge was conditioned with 10 mL of deionized water thrice. The purified oligonucleotide dissolved thoroughly in 2.5 mL deionized water was applied to the cartridge with very slow drop wise elution. The salt free oligomer was eluted with 3.5 mL deionized water directly into a screw cap vial. Final HPLC and Electrospray LC/MS Analysis [0089] Approximately 0.10 OD of oligomer is dissolved in water and then pipetted in special vials for IEX-HPLC and LC/MS analysis. Analytical HPLC and ES LC- MS established the integrity of the chimeric oligonucleotides. [0090] The cholesterol and tocopherol conjugated sequences were analyzed by high-performance liquid chromatography (HPLC) on a Luna C8 reverse-phase column. The buffers were 20 mM NaOAc in 10 % CH 3 CN (buffer A) and 20 mM NaOAc in 70% CH 3 CN (buffer B). Analytical HPLC and ES LC-MS established the integrity of the conjugated oligonucleotides Post Synthesis Conjugation: [0091] 5’-Folate conjugated ASOs.: To a solution of 5’-hexylamino ASO in 0.1 M sodium tetraborate buffer, pH 8.5 (2 mM) a solution of Folate-NHS ester (3 mole equivalent) dissolved in DMSO (40 mM) was added, and the reaction mixture was stirred at room temperature for 3 h. The Reaction mixture concentrated under reduced pressure. The residue was dissolved in water and purified by HPLC on a strong anion exchange column (GE Healthcare Bioscience, Source 30Q, 30 μm, 2.54 x 8 cm, A = 100 mM ammonium acetate in 30% aqueous CH3CN, B = 1.8 M NaBr in A, 0-60% of B in 60 min, flow 10 mL/min). The residue was desalted by in house packed Sephadex G-25 column to yield the 5’-Folate conjugated ASOs in an isolated yield of 62-80%. The folate conjugated ASOs were characterized by IEX-HPLC and Thermo Fischer ESI-LC-MS system. Table 6 depicts exemplary nucleic acids and structures. Table 6: Abbreviations for nucleic acid structures
[0092] Any of the structures shown in Table 6 can be combined with any base, thereby generating various combinations of structures. For example, using the abbreviations and structures from Table 6, one skilled in the art understands that the abbreviation “AmG”
represents “ .” Furthermore, additional structures not depicted in the tables, but described elsewhere throughout the application may be used and combined with any base described in the tables or elsewhere throughout the application. Pharmaceutical Compositions [0093] Some embodiments described herein relate to pharmaceutical compositions that comprise, consist essentially of, or consist of an effective amount of an oligonucleotide described herein and a pharmaceutically acceptable carrier, excipient, or combination thereof. A pharmaceutical composition described herein is suitable for human and/or veterinary applications. [0094] The terms “function” and “functional” as used herein refer to a biological, enzymatic, or therapeutic function. [0095] The terms “effective amount” or “effective dose” is used to indicate an amount of an active compound, or pharmaceutical agent, that elicits the biological or medicinal response indicated. For example, an effective amount of compound can be the amount needed to alleviate or ameliorate symptoms of disease or prolong the survival of the subject being treated This response may occur in a tissue, system, animal or human and includes alleviation of the signs or symptoms of the disease being treated. Determination of an effective amount is well within the capability of those skilled in the art, in view of the disclosure provided herein. The effective amount of the compounds disclosed herein required as a dose will depend on the route of administration, the type of animal, including human, being treated, and the physical characteristics of the specific animal under consideration. The dose can be tailored to achieve a desired effect, but will depend on such factors as weight, diet, concurrent medication and other factors which those skilled in the medical arts will recognize. [0096] The term “pharmaceutically acceptable salts” includes relatively non- toxic, inorganic and organic acid, or base addition salts of compositions, including without limitation, analgesic agents, therapeutic agents, other materials, and the like. Examples of pharmaceutically acceptable salts include those derived from mineral acids, such as hydrochloric acid and sulfuric acid, and those derived from organic acids, such as ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and the like. Examples of suitable inorganic bases for the formation of salts include the hydroxides, carbonates, and bicarbonates of ammonia, sodium, lithium, potassium, calcium, magnesium, aluminum, zinc, and the like. Salts may also be formed with suitable organic bases, including those that are non-toxic and strong enough to form such salts. For example, the class of such organic bases may include but are not limited to mono-, di-, and trialkylamines, including methylamine, dimethylamine, and triethylamine; mono-, di-, or trihydroxyalkylamines including mono-, di- , and triethanolamine; amino acids, including glycine, arginine and lysine; guanidine; N- methylglucosamine; N-methylglucamine; L-glutamine; N-methylpiperazine; morpholine; ethylenediamine; N-benzylphenethylamine; trihydroxymethyl aminoethane. [0097] “Formulation”, “pharmaceutical composition”, and “composition” as used interchangeably herein are equivalent terms referring to a composition of matter for administration to a subject. [0098] The term “pharmaceutically acceptable” means compatible with the treatment of a subject, and in particular, a human. [0099] The terms “agent” refers to an active agent that has biological activity and may be used in a therapy. Also, an “agent” can be synonymous with “at least one agent,” “compound,” or “at least one compound,” and can refer to any form of the agent, such as a derivative, analog, salt or a prodrug thereof. The agent can be present in various forms, components of molecular complexes, and pharmaceutically acceptable salts (e.g., hydrochlorides, hydrobromides, sulfates, phosphates, nitrates, borates, acetates, maleates, tartrates, and salicylates). The term “agent” can also refer to any pharmaceutical molecules or compounds, therapeutic molecules or compounds, matrix forming molecules or compounds, polymers, synthetic molecules and compounds, natural molecules and compounds, and any combination thereof. [0100] The term “subject” as used herein has its ordinary meaning as understood in light of the specification and refers to an animal that is the object of treatment, inhibition, or amelioration, observation or experiment. “Animal” has its ordinary meaning as understood in light of the specification and includes cold- and warm-blooded vertebrates and/or invertebrates such as fish, shellfish, or reptiles and, in particular, mammals. “Mammal” has its ordinary meaning as understood in light of the specification, and includes but is not limited to mice, rats, rabbits, guinea pigs, dogs, cats, sheep, goats, cows, horses, primates, such as humans, monkeys, chimpanzees, or apes. In some embodiments, the subject is human. [0101] Proper formulation is dependent upon the route of administration chosen. Techniques for formulation and administration of the compounds described herein are known to those skilled in the art. Multiple techniques of administering a compound exist in the art including, but not limited to, enteral, oral, rectal, topical, sublingual, buccal, intraaural, epidural, epicutaneous, aerosol, parenteral delivery, including intramuscular, subcutaneous, intra-arterial, intravenous, intraportal, intra-articular, intradermal, peritoneal, intramedullary injections, intrathecal, direct intraventricular, intraperitoneal, intranasal or intraocular injections. Pharmaceutical compositions will generally be tailored to the specific intended route of administration. Pharmaceutical compositions can also be administered to isolated cells from a patient or individual, such as T cells, Natural Killer cells, B cells, macrophages, lymphocytes, stem cells, bone marrow cells, or hematopoietic stem cells. [0102] The pharmaceutical compound can also be administered in a local rather than systemic manner, for example, via injection of the compound directly into an organ, tissue, cancer, tumor or infected area, often in a depot or sustained release formulation. Furthermore, one may administer the compound in a targeted drug delivery system, for example, in a liposome coated with a tissue specific antibody. The liposomes may be targeted to and taken up selectively by the organ, tissue, cancer, tumor, or infected area. [0103] The pharmaceutical compositions disclosed herein may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or tableting processes. As described herein, compounds used in a pharmaceutical composition may be provided as salts with pharmaceutically compatible counterions. [0104] As used herein, a “carrier” refers to a compound, particle, solid, semi- solid, liquid, or diluent that facilitates the passage, delivery and/or incorporation of a compound to cells, tissues and/or bodily organs. For example, without limitation, a lipid nanoparticle (LNP) is a type of carrier that can encapsulate an oligonucleotide to thereby protect the oligonucleotide from degradation during passage through the bloodstream and/or to facilitate delivery to a desired organ, such as to the liver. [0105] As used herein, a “diluent” refers to an ingredient in a pharmaceutical composition that lacks pharmacological activity but may be pharmaceutically necessary or desirable. For example, a diluent may be used to increase the bulk of a potent drug whose mass is too small for manufacture and/or administration. It may also be a liquid for the dissolution of a drug to be administered by injection, ingestion or inhalation. A common form of diluent in the art is a buffered aqueous solution such as, without limitation, phosphate buffered saline that mimics the composition of human blood. [0106] The term “excipient” has its ordinary meaning as understood in light of the specification, and refers to inert substances, compounds, or materials added to a pharmaceutical composition to provide, without limitation, bulk, consistency, stability, binding ability, lubrication, disintegrating ability etc., to the composition. Excipients with desirable properties include but are not limited to preservatives, adjuvants, stabilizers, solvents, buffers, diluents, solubilizing agents, detergents, surfactants, chelating agents, antioxidants, alcohols, ketones, aldehydes, ethylenediaminetetraacetic acid (EDTA), citric acid, salts, sodium chloride, sodium bicarbonate, sodium phosphate, sodium borate, sodium citrate, potassium chloride, potassium phosphate, magnesium sulfate sugars, dextrose, fructose, mannose, lactose, galactose, sucrose, sorbitol, cellulose, serum, amino acids, polysorbate 20, polysorbate 80, sodium deoxycholate, sodium taurodeoxycholate, magnesium stearate, octylphenol ethoxylate, benzethonium chloride, thimerosal, gelatin, esters, ethers, 2-phenoxyethanol, urea, or vitamins, or any combination thereof. The amount of the excipient may be found in a pharmaceutical composition at a percentage of 0%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 100% w/w or any percentage by weight in a range defined by any two of the aforementioned numbers. [0107] The term “adjuvant” as used herein refers to a substance, compound, or material that stimulates the immune response and increase the efficacy of protective immunity and is administered in conjunction with an immunogenic antigen, epitope, or composition. Adjuvants serve to improve immune responses by enabling a continual release of antigen, up-regulation of cytokines and chemokines, cellular recruitment at the site of administration, increased antigen uptake and presentation in antigen presenting cells, or activation of antigen presenting cells and inflammasomes. Commonly used adjuvants include but are not limited to alum, aluminum salts, aluminum sulfate, aluminum hydroxide, aluminum phosphate, calcium phosphate hydroxide, potassium aluminum sulfate, oils, mineral oil, paraffin oil, oil-in-water emulsions, detergents, MF59®, squalene, AS03, Į- tocopherol, polysorbate 80, AS04, monophosphoryl lipid A, virosomes, nucleic acids, polyinosinic:polycytidylic acid, saponins, QS-21, proteins, flagellin, cytokines, chemokines, IL-1, IL-2, IL-12, IL-15, IL-21, imidazoquinolines, CpG oligonucleotides, lipids, phospholipids, dioleoyl phosphatidylcholine (DOPC), trehalose dimycolate, peptidoglycans, bacterial extracts, lipopolysaccharides, or Freund’s Adjuvant, or any combination thereof. [0108] The term “purity” of any given substance, compound, or material as used herein refers to the actual abundance of the substance, compound, or material relative to the expected abundance. For example, the substance, compound, or material may be at least 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% pure, including all decimals in between. Purity may be affected by unwanted impurities, including but not limited to side products, isomers, enantiomers, degradation products, solvent, carrier, vehicle, or contaminants, or any combination thereof. Purity can be measured technologies including but not limited to chromatography, liquid chromatography, gas chromatography, spectroscopy, UV-visible spectrometry, infrared spectrometry, mass spectrometry, nuclear magnetic resonance, gravimetry, or titration, or any combination thereof. Methods of Use [0109] Some embodiments disclosed herein related to selecting a subject or patient in need. In some embodiments, a patient is selected who is in need of treatment, inhibition, amelioration, prevention or slowing of diseases or conditions associated with PD- L1 dysregulation. In some embodiments, such diseases or conditions associated with PD-L1 dysregulation may include, for example, cancer, HCC, viral infections, or HBV. In some embodiments, a patient is selected who has previously been treated for the disease or disorder described herein. In some embodiments, a patient is selected who has previously been treated for being at risk for the disease or disorder described herein. In some embodiments, a patient is selected who has developed a recurrence of the disease or disorder described herein. In some embodiments, a patient is selected who has developed resistance to therapies for the disease or disorder described herein. In some embodiments, a patient is selected who may have any combination of the aforementioned selection criteria. [0110] Compounds disclosed herein can be evaluated for efficacy and toxicity using known methods. A non-limiting list of potential advantages of an oligonucleotide described herein include improved stability, increased safety profile, increased efficacy, increased binding to the target, increased specificity for the target (for example, a cancer cell or virally infected cell). [0111] The terms “treating,” “treatment,” “therapeutic,” or “therapy” as used herein has its ordinary meaning as understood in light of the specification, and do not necessarily mean total cure or abolition of the disease or condition. The term “treating” or “treatment” as used herein (and as well understood in the art) also means an approach for obtaining beneficial or desired results in a subject's condition, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of the extent of a disease, stabilizing (i.e., not worsening) the state of disease, prevention of a disease's transmission or spread, delaying or slowing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission, whether partial or total and whether detectable or undetectable. “Treating” and “treatment” as used herein also include prophylactic treatment. Treatment methods comprise administering to a subject a therapeutically effective amount of an active agent. The administering step may consist of a single administration or may comprise a series of administrations. The compositions are administered to the subject in an amount and for a duration sufficient to treat the patient. The length of the treatment period depends on a variety of factors, such as the severity of the condition, the age and genetic profile of the patient, the concentration of active agent, the activity of the compositions used in the treatment, or a combination thereof. It will also be appreciated that the effective dosage of an agent used for the treatment or prophylaxis may increase or decrease over the course of a particular treatment or prophylaxis regime. Changes in dosage may result and become apparent by standard diagnostic assays known in the art. In some instances, chronic administration may be required. [0112] Some embodiments described herein relate to a method of treating, inhibiting, ameliorating, preventing, or slowing the disease or disorder described herein. In some embodiments, the methods include administering to a subject identified as suffering from the disease or disorder described herein an effective amount of an ASO described herein, or a pharmaceutical composition that includes an effective amount of an ASO as described herein. Other embodiments described herein relate to using an ASO as described herein in the manufacture of a medicament for treating, inhibiting ameliorating, preventing, or slowing the disease or disorder described herein. Still other embodiments described herein relate to the use of an ASO as described herein or a pharmaceutical composition that includes an effective amount of an ASO as described herein for treating, inhibiting ameliorating, preventing, or slowing the disease or disorder described herein. [0113] Some embodiments described herein relate to a method for inhibiting replication of a cancer cell or a virus that can include contacting the cell or virus or administering to a subject identified as suffering from a cancer or a viral infection with an effective amount of an ASO described herein, or a pharmaceutical composition that includes an effective amount of an ASO described herein. Other embodiments described herein relate to the use of an effective amount of an ASO described herein, or a pharmaceutical composition that includes an effective amount of an ASO described herein in the manufacture of a medicament for inhibiting replication of a cancer cell or virus. Still other embodiments described herein relate to an effective amount of an ASO described herein, or a pharmaceutical composition that includes an effective amount of an ASO described herein for inhibiting replication of a cancer cell or virus. In some embodiments, the cancer cell is an HCC cell. In some embodiments, the virus is hepatitis B. [0114] Some embodiments described herein relate to a method for inhibiting cell proliferation, such as inhibiting cell proliferation of a cancer cell or cell infected with a virus, that can include administering to a subject identified as suffering from a disease wherein inhibiting cell proliferation is desirable with an effective amount of an ASO described herein, or a pharmaceutical composition that includes effective amount of an ASO described herein. Other embodiments described herein relate to the use of an effective amount of an oligonucleotide described herein, or a pharmaceutical composition that includes an effective amount of an ASO described herein in the manufacture of a medicament for inhibiting cell proliferation, such as inhibiting cell proliferation of a cancer cell or cell infected with a virus. Still other embodiments described herein relate to an effective amount of an ASO described herein, or a pharmaceutical composition that includes an effective amount of an ASO described herein for inhibiting cell proliferation, such as inhibiting cell proliferation of a cancer cell or cell infected with a virus. In some embodiments, the cancer cell is an HCC cell. In some embodiments, the cell infected with a virus is infected with hepatitis B virus. [0115] Some embodiments described herein relate to a method of inducing apoptosis of a cell (for example, a cancer cell or cell infected with a virus) that can include contacting the cell with an effective amount of an ASO described herein, or a pharmaceutical composition that includes an effective amount of an ASO as described herein. Other embodiments described herein relate to using an effective amount of an ASO as described herein or a pharmaceutical composition that includes an effective amount of an ASO in the manufacture of a medicament for inducing apoptosis of a cell, such as a cancer cell or cell infected with a virus. Still other embodiments described herein relate to the use of an effective amount of an ASO as described herein or a pharmaceutical composition that includes an effective amount of an ASO as described herein for inducing apoptosis of a cell, such as a cancer cell or cell infected with a virus. In some embodiments, the cancer cell is an HCC cell. In some embodiments, the cell infected with a virus is infected with hepatitis B virus. [0116] Some embodiments described herein relate to a method of decreasing the viability of a cell (for example, a cancer cell or cell infected with a virus) that can include contacting the cell with an effective amount of an ASO described herein, or a pharmaceutical composition that includes an effective amount of an ASO as described herein. Other embodiments described herein relate to using an ASO as described herein in the manufacture of a medicament for decreasing the viability of a cell, such as a cancer cell or cell infected with a virus. Still other embodiments described herein relate to the use of an effective amount of an ASO as described herein or a pharmaceutical composition that includes an effective amount of an ASO as described herein for decreasing the viability of a cell, such as a cancer cell or cell infected with a virus. In some embodiments, the cancer cell is an HCC cell. In some embodiments, the cell infected with a virus is infected with hepatitis B virus. [0117] In some embodiments, the effective amount of an ASO for a human subject is 1, 10, 50, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000 ^g, or 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 300, 400, 500, 600, 700, 800, 900, 1000 mg or any amount within the range defined by any two aforementioned amounts. In some embodiments, the effective amount of an ASO for a human subject is 1, 10, 50, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000 ng/kg, or 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 300, 400, 500, 600, 700, 800, 900, 1000 ^g/kg or any amount within the range defined by any two aforementioned amounts. In some embodiments, the effective amount of an ASO is dosed more than one time. In some embodiments, the ASO dose is administered every 1, 2, 3, 4, 5, 6, 7 days, or 1, 2, 3, 4 weeks, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months, or 1, 2, 3, 4, 5 years, or any period or combination thereof within the range defined by any two aforementioned times. In some embodiments, at least one loading dose and at least one maintenance dose is administered to the subject, where the at least one loading dose is a higher dose of the ASO than the at least one maintenance dose. [0118] As used herein, the term “combination therapy” is intended to define therapies which comprise the use of a combination of two or more pharmaceutical compounds/agents or therapies. Thus, references to “combination therapy”, “combinations” and the use of compounds/agents “in combination” in this application may refer to compounds/agents that are administered as part of the same overall treatment regimen. As such, the dosage or timing of each of the two or more compounds/agents may differ: each may be administered at the same time or at different times. Accordingly, the compounds/agents of the combination may be administered sequentially (e.g. before or after) or simultaneously, either in the same pharmaceutical formulation (i.e. together), or in different pharmaceutical formulations (i.e. separately). Each of the two or more compounds/agents in a combination therapy may also differ with respect to the route of administration. [0119] The term “inhibitor”, as used herein, refers to an enzyme inhibitor or receptor inhibitor which is a molecule that binds to an enzyme or receptor, and decreases and/or blocks its activity. The term may relate to a reversible or an irreversible inhibitor. [0120] Cancer may be treated with surgery, radiation therapy, chemotherapy, targeted therapies, immunotherapy or hormonal therapies. Any of these mentioned therapies may be used in conjunction with another therapy as a combination therapy. Chemotherapeutic compounds include but are not limited to alemtuzumab, altretamine, azacitidine, bendamustine, bleomycin, bortezomib, busulfan, cabazitaxel, capecitabine, carboplatin, carmofur, carmustine, chlorambucil, chlormethine, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin, decitabine, denosumab, docetaxel, doxorubicin, epirubicin, estramustine, etoposide, everolimus, floxuridine, fludarabine, fluorouracil, fotemustine, gemcitabine, gemtuzumab, hydroxycarbamide, ibritumomab, idarubicin, ifosfamide, irinotecan, ixabepilone, lomustine, melphalan, mercaptopurine, methotrexate, mitomycin, mitoxantrone, nedaplatin, nelarabine, ofatumumab, oxaliplatin, paclitaxel, pemetrexed, pentostatin, pertuzumab, procarbazine, raltitrexed, streptozotocin, tegafur, temozolomide, temsirolimus, teniposide, tioguanine, topotecan, tositumomab, valrubicin, vinblastine, vincristine, vindesine, vinflunine, or vinorelbine, or any combination thereof. [0121] As used herein, the term “protein kinase inhibitor” refers to inhibitors of protein kinases, serine/threonine kinases, tyrosine kinases, or dual-specificity kinases for the treatment of cancer or other illness. In some embodiments, the protein kinase inhibitor is a small molecule, compound, polysaccharide, lipid, peptide, polypeptide, protein, antibody, nucleoside, nucleoside analog, nucleotide, nucleotide analog, nucleic acid, or oligonucleotide. In some embodiments, the protein kinase inhibitor includes but is not limited to acalabrutinib, adavosertib, afatinib, alectinib, axitinib, binimetinib, bosutinib, brigatinib, cediranib, ceritinib, cetuximab, cobimetinib, crizotinib, cabozantinib, dacomitinib, dasatinib, entrectinib, erdafitinib, erlotinib, fostamatinib, gefitinib, ibrutinib, imatinib, lapatinib, lenvatinib, lestaurtinib, lortatinib, masitinib, momelotinib, mubritinib, neratinib, nilotinib, nintedanib, olmutinib, osimertinib, pacritinib, panitumumab, pazopanib, pegaptanib, ponatinib, radotinib, regorafenib, rociletinib, ruxolitinib, selumetinib, semaxanib, sorafenib, sunitinib, SU6656, tivozanib, toceranib, trametinib, trastuzumab, vandetanib, or vemurafenib, or any combination thereof. [0122] As used herein, the term “checkpoint inhibitor” refers to an immunotherapy that targets immune checkpoints to stimulate immune function. In some embodiments, the checkpoint inhibitor is a small molecule, compound, polysaccharide, lipid, peptide, polypeptide, protein, antibody, nucleoside, nucleoside analog, nucleotide, nucleotide analog, nucleic acid, or oligonucleotide. In some embodiments, the immune checkpoint is the PD-1/PD-L1 checkpoint. In some embodiments, the PD-1 checkpoint includes but is not limited to nivolumab, pembrolizumab, spartalizumab, cemiplimab, camrelizumab, sintilimab, tislelizumab, toripalimab, AMP-224 or AMP-514, or any combination thereof. In some embodiments, the PD-L1 checkpoint inhibitor includes but is not limited to atezolizumab, avelumab, durvalumab, KN035, AUNP12, CA-170, or BMS-986189, or any combination thereof. In some embodiments, the immune checkpoint is the CTLA-4 checkpoint. In some embodiments, the CTLA-4 checkpoint inhibitor includes but is not limited to ipilimumab or tremilimumab, or any combination thereof. [0123] As used herein, the term “VEGF inhibitor” refers to inhibitors of vascular endothelial growth factor (VEGF) or a VEGF receptor (VEGFR). In some embodiments, the VEGF inhibitor is a small molecule, compound, polysaccharide, lipid, peptide, polypeptide, protein, antibody, nucleoside, nucleoside analog, nucleotide, nucleotide analog, nucleic acid, or oligonucleotide. In some embodiments, the VEGF inhibitor includes but is not limited to aflibercept, axitinib, bevacizumab, brivanib, cabozantinib, cediranib, lenvatinib, linifinib, nintedanib, pazopanib, ponatinib, ramucirumab, regorafenib, semaxanib, sorafenib, sunitinib, tivozanib, toceranib, or vandetanib, or any combination thereof. [0124] As used herein, the term “antiviral medication” refers to a pharmaceutical composition administered to treat a viral infection. In some embodiments, the viral infection is caused by adenovirus, Ebola virus, coronavirus, Epstein-Barr virus (EBV), Friend virus, hantavirus, hepatitis B virus (HBV), hepatitis C virus (HCV), herpes simplex virus, human immunodeficiency virus (HIV), human metapneumovirus, human papillomavirus (HPV), influenza virus, Japanese encephalitis virus, Kaposi’s sarcoma-associated herpesvirus, lymphocytic choriomeningitis virus, parainfluenza virus, rabies virus, respiratory syncytial virus, rhinovirus, varicella zoster virus. In some embodiments, the antiviral medication is a small molecule, compound, polysaccharide, lipid, peptide, polypeptide, protein, antibody, nucleoside, nucleoside analog, nucleotide, nucleotide analog, nucleic acid, or oligonucleotide. In some embodiments, the antiviral medication is an interferon, a capsid assembly modulator, a sequence specific oligonucleotide, an entry inhibitor, or a small molecule immunomodulatory. In some embodiments, the antiviral medication includes but is not limited to AB-423, AB-506, ABI-H2158, ABI-HO731, acyclovir, adapromine, adefovir, alafenamide, amantadine, asunaprevir, baloxavir marboxil, beclabuvir, boceprevir, brivudine, cidofovir, ciluprevir, clevudine, cytarabine, daclatasvir, danoprevir, dasabuvir, deleobuvir, dipivoxil, edoxudine, elbasvir, entecavir, faldaprevir, famciclovir, favipiravir, filibuvir, fomivirsen, foscarnet, galidesivir, ganciclovir, glecaprevir, GLS4, grazoprevir, idoxuridine, imiquimod, IFN-Į, interferon alfa 2b, JNJ-440, JNJ-6379, lamivudine, laninamivir, ledipasvir, mericitabine, methisazone, MK-608, moroxydine, narlaprevir, NITD008, NZ-4, odalasvir, ombitasvir, oseltamivir, paritaprevir, peginterferon alfa-2a, penciclovir, peramivir, pibrentasvir, pimodivir, pleconaril, podophyllotoxin, presatovir, radalbuvir, ravidasvir, remdesivir, REP 2139, REP 2165, resiquimod, RG7907, ribavirin, rifampicin, rimantadine, ruzasvir, samatasvir, setrobuvir, simeprevir, sofosbuvir, sorivudine, sovaprevir, taribavirin, telaprevir, telbivudine, tenofovir, tenofovir disoproxil, triazavirin, trifluridine, tromantadine, umifenovir, uprifosbuvir, valaciclovir, valgancicovir, vaniprevir, vedroprevir, velpatasvir, vidarabine, voxilaprevir, or zanamivir, or any combination thereof. [0125] The term “% w/w” or “% wt/wt” as used herein has its ordinary meaning as understood in light of the specification and refers to a percentage expressed in terms of the weight of the ingredient or agent over the total weight of the composition multiplied by 100. The term “% v/v” or “% vol/vol” as used herein has its ordinary meaning as understood in the light of the specification and refers to a percentage expressed in terms of the liquid volume of the compound, substance, ingredient, or agent over the total liquid volume of the composition multiplied by 100. [0126] The invention is generally disclosed herein using affirmative language to describe the numerous embodiments. The invention also includes embodiments in which subject matter is excluded, in full or in part, such as substances or materials, method steps and conditions, protocols, or procedures. EXAMPLES [0127] Some aspects of the embodiments discussed above are disclosed in further detail in the following examples, which are not in any way intended to limit the scope of the present disclosure. Those in the art will appreciate that many other embodiments also fall within the scope of the invention, as it is described herein above and in the claims. Example 1: Antisense oligonucleotide (ASO) design [0128] ASOs were selected having 14-20 nucleotides in length. To account for optimal gapmer design, mismatches were allowed only in the outer flank regions of the ASO, as shown in Table 7. The typical DNA nucleotide central gap was 8, 9 or 10 nucleotides in length. Therefore, the central 10 or 11 nucleotides were kept fully conserved. Table 7 summarizes the allowed mismatch position for ASOs of different lengths. ASOs of 14 or 15 nucleotides in length were not allowed to have mismatches. ASOs of 16 or 17 nucleotides in length included 1 or fewer mismatches within the three outer RNA nucleotides on either 5’ or 3’ end. ASOs of 18 or 19 nucleotides in length included 2 or fewer mismatches within the four outer RNA nucleotides on either 5’ or 3’ end. ASOs of 20 nucleotides in length included 2 or fewer mismatches within the five outer RNA nucleotides on either 5’ or 3’ end. Table 7: Allowed mismatch positions on ASOs Example 2: ASOs targeting the human CD274 gene (PD-L1) [0129] ASOs were designed using the human CD274 mRNA transcript (NCBI accession number NM_014143.4, 3634 nt in length, SEQ ID NO: 1) as the template.14-mers are depicted in Table 8 (SEQ ID NOs: 2-28). 15-mers are depicted in Table 9 (SEQ ID NOs: 29-46). 16-mers are depicted in Table 10 (SEQ ID NOs: 47-86 and 240-301). 17-mers are depicted in Table 11 (SEQ ID NOs: 87-114). 18-mers are depicted in Table 12 (SEQ ID NOs: 115-165). 19-mers are depicted in Table 13 (SEQ ID NOs: 166-204). 20-mers are depicted in Table 14 (SEQ ID NO: 205-239). [0130] Any of the ASOs listed here, and the individual nucleobases, sugars, linkages, nucleosides, nucleotides and additional moieties thereof, can be constructed and used with any of the modifications described herein. The sequences listed in Tables 8-14 and SEQ ID NOs: 2-301 represent the unmodified oligonucleotide sequence prior to application of modifications. Table 8: CD274 ASOs – 14-mers
Table 9: CD274 ASOs – 15mers Table 10: CD274 ASOs – 16mers
Table 11: CD274 ASOs – 17mers
Table 12: CD274 ASOs – 18mers Table 13: CD274 ASOs – 19mers
Table 14: CD274 ASOs – 20mers
Example 3: Treatment of cancer using CD274 ASOs [0131] A human patient presents with a cancer, such as a hepatocellular carcinoma (HCC). The cancer is a non-metastatic or metastatic cancer. In the case of HCC, the patient may also have another liver condition, such as fibrosis, cirrhosis, non-alcoholic liver disease, hepatitis, hepatitis B, or hepatitis C. An effective amount of a CD274 ASO or a pharmaceutical composition comprising an effective amount of a CD274 ASO is administered to the patient parenterally. The CD274 ASO is selected from the group consisting of SEQ ID NOs: 2-301. The CD274 ASO can optionally have any of the modifications to individual nucleobases, sugars, linkages, nucleosides, or nucleotides as described herein. The CD274 ASO can also optionally have a covalently conjugated targeting moiety to improve selectivity to tumor and/or liver tissue. The CD274 ASO can be constructed of deoxyribose sugars (DNA nucleotides), ribose sugars (RNA nucleotides) or any combination thereof The CD274 ASO can be constructed of unmodified nucleotides or modified nucleotides or any combination thereof and optionally can be a gapmer, mixmer, or blockmer. The CD274 ASO or pharmaceutical composition comprising the CD274 ASO can optionally be administered as a combination therapy with another anti-neoplastic compound or therapy. [0132] Following administration of an effective amount of the CD274 ASO or the pharmaceutical composition comprising an effective amount of the CD274 ASO, the cancer is reduced or eliminated. Example 4: Treatment of hepatitis B using CD274 ASOs [0133] A human patient presents with a hepatitis B infection. The hepatitis B infection is acute or chronic. The hepatitis B infection may also be coincidental with a hepatitis D infection. The patient may also have another liver conditions, such as fibrosis, cirrhosis, non-alcoholic liver disease, or HCC. An effective amount of a CD274 ASO or a pharmaceutical composition comprising an effective amount of a CD274 ASO is administered to the patient parenterally. The CD274 ASO is selected from the group consisting of SEQ ID NOs: 2-301. The CD274 ASO can optionally have any of the modifications to individual nucleobases, sugars, linkages, nucleosides, or nucleotides as described herein. The CD274 ASO can also optionally have a covalently conjugated targeting moiety to improve selectivity to liver tissue. The CD274 ASO can be constructed of deoxyribose sugars (DNA nucleotides), ribose sugars (RNA nucleotides) or any combination thereof The CD274 ASO can be constructed of unmodified nucleotides or modified nucleotides or any combination thereof and optionally can be a gapmer, mixmer, or blockmer. The CD274 ASO or pharmaceutical composition comprising the CD274 ASO can optionally be administered as a combination therapy with another antiviral medication. [0134] Following administration of an effective amount of the CD274 ASO or the pharmaceutical composition comprising an effective amount of the CD274 ASO, the hepatitis B infection (and optionally, hepatitis D infection) is reduced or eliminated. Example 5: Treatment of hepatocellular carcinoma cells using ASOs [0135] Human hepatocellular carcinoma cells (SNU-387) were seeded at 30,000 cells/well in a 96-well plate. The ASOs, including any of SEQ ID NOs: 2-301, were transfected with Lipofectamine RNAiMax (Life Technologies) in the seeded SNU-387 cells. The ASOs included any of the modifications described herein, including modification of individual nucleobases, sugars, linkages, nucleosides, or nucleotides. ASOs were screened at two concentrations, 100 nM and 1 nM or 50 nM and 1 nm. Active ASOs were further screened to obtain EC50 values via dose response curves. Selected ASO subsets were selected having greater than 50% K.D. at 100 nM, and limited toxicity. [0136] For dose response curves, a 3-fold dilution series of ASO (top dose 50 nM; six or eight concentrations tested total) was tested. The cells were harvested 48 hours after transfection, and RNA was extracted with RNeasy Kit (Qiagen). RT-qPCR was performed to asses PD-L1 gene knockdown. Cell viability was assessed at 48 hours post transfection using CCK8 assay. Data was fit with fitting software using a four-parameter dose response equation. Table 15 provides representative EC50 and CC50 values for selected ASOs, each of which have a LNA-DNA-LNA (3-10-3) modification with each linkage between the nucleosides a phosphorothioate (a PS backbone), and with all cytosines a (5m)C. Tables 16 and 17 depict percent reduction of PD-L1 gene for select ASOs, each of which have a LNA-DNA-LNA (3-10-3) modification. Figure 1 depicts the percent PD-L1 knockdown as a function of log concentration of ASO in nM. Figure 2 depicts the fraction of PD-L1 mRNA remaining for three exemplary ASOs as a function of log concentration of ASO in nM. Table 15: EC50 and CC50 for select ASOs A ≤ 1 nM; B > 1-5 nM; C > 5-10 nM X > 50 nM; Y ≤ 50 nM Table 16: Percent Reduction of PD-L1 Gene with select ASOs A > 75%-100%; B > 50%-75%; C > 25%-50%; D = 0%-25% X > 100 nM; Y ≤100 nM Table 17: Percent Reduction of PD-L1 Gene with select ASOs A > 75%-100%; B > 50%-75%; C > 25%-50%; D = 0-25% X > 50 nM; Y ≤ 50 nM [0137] The example ASOs, including the example sequences and example modifications as described in the examples, are intended as exemplary sequences and modifications. However, it is to be understood that the disclosure relates to any ASO sequence as set forth herein, having any modification or combination of modifications as set forth herein may be implemented in the examples. [0138] In at least some of the previously described embodiments, one or more elements used in an embodiment can interchangeably be used in another embodiment unless such a replacement is not technically feasible. It will be appreciated by those skilled in the art that various other omissions, additions and modifications may be made to the methods and structures described above without departing from the scope of the claimed subject matter. All such modifications and changes are intended to fall within the scope of the subject matter, as defined by the appended claims. [0139] With respect to the use of substantially any plural and/or singular terms herein, those having skill in the art can translate from the plural to the singular and/or from the singular to the plural as is appropriate to the context and/or application. The various singular/plural permutations may be expressly set forth herein for sake of clarity. [0140] It will be understood by those within the art that, in general, terms used herein, and especially in the appended claims (e.g., bodies of the appended claims) are generally intended as “open” terms (e.g., the term “including” should be interpreted as “including but not limited to,” the term “having” should be interpreted as “having at least,” the term “includes” should be interpreted as “includes but is not limited to,” etc.). It will be further understood by those within the art that if a specific number of an introduced claim recitation is intended, such an intent will be explicitly recited in the claim, and in the absence of such recitation no such intent is present. For example, as an aid to understanding, the following appended claims may contain usage of the introductory phrases “at least one” and “one or more” to introduce claim recitations. However, the use of such phrases should not be construed to imply that the introduction of a claim recitation by the indefinite articles “a” or “an” limits any particular claim containing such introduced claim recitation to embodiments containing only one such recitation, even when the same claim includes the introductory phrases “one or more” or “at least one” and indefinite articles such as “a” or “an” (e.g., “a” and/or “an” should be interpreted to mean “at least one” or “one or more”); the same holds true for the use of definite articles used to introduce claim recitations. In addition, even if a specific number of an introduced claim recitation is explicitly recited, those skilled in the art will recognize that such recitation should be interpreted to mean at least the recited number (e.g., the bare recitation of “two recitations,” without other modifiers, means at least two recitations, or two or more recitations). Furthermore, in those instances where a convention analogous to “at least one of A, B, and C, etc.” is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., “a system having at least one of A, B, and C” would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and/or A, B, and C together, etc.). In those instances where a convention analogous to “at least one of A, B, or C, etc.” is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., “ a system having at least one of A, B, or C” would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and/or A, B, and C together, etc.). It will be further understood by those within the art that virtually any disjunctive word and/or phrase presenting two or more alternative terms, whether in the description or claims, should be understood to contemplate the possibilities of including one of the terms, either of the terms, or both terms. For example, the phrase “A or B” will be understood to include the possibilities of “A” or “B” or “A and B.” [0141] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group. [0142] As will be understood by one skilled in the art, for any and all purposes, such as in terms of providing a written description, all ranges disclosed herein also encompass any and all possible sub-ranges and combinations of sub-ranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non- limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like include the number recited and refer to ranges which can be subsequently broken down into sub-ranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 articles refers to groups having 1, 2, or 3 articles. Similarly, a group having 1-5 articles refers to groups having 1, 2, 3, 4, or 5 articles, and so forth. [0143] While various aspects and embodiments have been disclosed herein, other aspects and embodiments will be apparent to those skilled in the art. The various aspects and embodiments disclosed herein are for purposes of illustration and are not intended to be limiting, with the true scope and spirit being indicated by the following claims. [0144] All references cited herein, including but not limited to published and unpublished applications, patents, and literature references, are incorporated herein by reference in their entirety and are hereby made a part of this specification. To the extent publications and patents or patent applications incorporated by reference contradict the disclosure contained in the specification, the specification is intended to supersede and/or take precedence over any such contradictory material. References [0145] 1. U.S. 2017/0283496 [0146] 2. Akinleye, A & Rasool Z. Immune Checkpoint Inhibitors of PD-L1 as Cancer Therapeutics. J. Hematol. Oncol. (2019) 12(1):92. [0147] 3. Wu, Y et al. PD-L1 Distribution and Perspective for Cancer Immunotherapy – Blockade, Knockdown, or Inhibition. Front. Immunol. (2019) 10:2022. [0148] 4. Sun, C et al. Regulation and Function of the PD-L1 Checkpoint. Immunity. (2018) 48(3):434-452. [0149] 5. Schönrich, G & Raferty MJ. The PD-1/PD-L1 Axis and Virus Infections: A Delicate Balance. Front. Cell. Infect. Microbiol. (2019) 9:207 [0150] 6. Østergaard, ME et al. Fluorinated Nucleotide Modifications Modulate Allele Selectivity of SNP-Targeting Antisense Oligonucleotides. Mol. Ther. Nucleic Acids. (2017) 7:20-30. [0151] 7. Di Fusco, D et al. Antisense Oligonucleotide: Basic Concepts and Therapeutic Application in Inflammatory Bowel Disease. Front Pharmacol. (2019) 10:305. [0152] 8. Wurster, CD & Ludolph AC. Antisense Oligonucleotides in Neurological Disorders. Ther. Adv. Neurol. Disord. (2018) 11:1-19. [0153] 9. Balsitis S et al. Safety and Efficacy of Anti-PD-L1 Therapy in the Woodchuck Model of HBV Infection. (2018) 13(2):1-23. [0154] Although the foregoing has been described in some detail by way of illustrations and examples for purposes of clarity and understanding, it will be understood by those of skill in the art that numerous and various modifications can be made without departing from the spirit of the present disclosure. Therefore, it should be clearly understood that the forms disclosed herein are illustrative only and are not intended to limit the scope of the present disclosure, but rather to also cover all modification and alternatives coming with the true scope and spirit of the invention.
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