Modified short interfering nucleic acid (SINA) molecules and uses thereof

Abstract

Disclosed herein are short interfering nucleic acid (siNA) molecules containing modified nucleotides and their uses. The siNA molecules can be double-stranded and can contain modified nucleotides selected from 2'-O-methyl nucleotides and 2'-fluoro nucleotides. Further disclosed herein are siNA molecules containing additional modifications, including phosphorylation blockers, conjugate moieties, or 5'-stabilized end caps. The siNA molecules can reduce or inhibit the production of hydroxysteroid dehydrogenase.

Description

[Technical Field]

[0001] This application claims priority to U.S. Provisional Patent Application No. 63 / 241,940, filed September 8, 2021, entitled "MODIFIED SHORT INTERFERING NUCLEIC ACID (SINA) MOLECULES AND USES THEREOF," which is hereby incorporated by reference in its entirety for all purposes.

[0002] Described are short interfering nucleic acid (siNA) molecules containing modified nucleotides, compositions and their uses. [Background technology]

[0003] RNA interference (RNAi) is a biological response to double-stranded RNA that mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids and regulates the expression of protein-coding genes. Short interfering nucleic acids (siNAs), such as siRNAs, have been developed for RNAi therapy to treat various diseases. For example, RNAi therapy has been proposed for the treatment of metabolic diseases, neurodegenerative diseases, cancer, and pathogenic infections (see, e.g., Rondindone, Biotechniques, 2018, 40(4S), doi.org / 10.2144 / 000112163; Boudreau and Davidson, Curr Top Dev Biol, 2006, 75:73-92; Chalbatani et al., Int J Nanomedicine, 2019, 14:3111-3128; Arbuthnot, Drug News Perspect, 2010, 23(6):341-50; and Chernikov et. al., Front. Pharmacol., 2019, doi.org / 10.3389 / fphar.2019.00444; each of which is incorporated by reference in its entirety). However, the main limitations of RNAi therapy are the ability to effectively deliver siRNA to target cells and siRNA degradation.

[0004] Nonalcoholic fatty liver disease (NAFLD) is an emerging global health problem and a potential risk factor for type 2 diabetes, cardiovascular disease, and chronic kidney disease. Nonalcoholic steatohepatitis (NASH), an advanced form of NAFLD, predisposes to the development of liver cirrhosis and hepatocellular carcinoma. The increasing prevalence of NASH highlights the need for novel therapeutic approaches. 17β-hydroxysteroid dehydrogenase type 13, also known as 17β-HSD13 (or HSD17B13), is an enzyme enriched in hepatocytes and localized in intracellular lipid droplets. HSD17B13 is significantly upregulated in the liver of patients with NAFLD and NASH and promotes lipogenesis. The role of HSD17B13 in lipogenesis appears to be mediated by its retinoid dehydrogenase activity. Reduction of HSD17B13 protein levels may result in reduction of ALT and AST levels, and may result in improvement of liver histology in NAFLD and NASH. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Rondindone, Biotechniques, 2018, 40(4S), doi.org / 10.2144 / 000112163 [Non-patent document 2] Boudreau and Davidson, Curr Top Dev Biol, 2006, 75:73-92 [Non-patent document 3] Chalbatani et al., Int J Nanomedicine, 2019, 14:3111-3128 [Non-patent document 4] Arbuthnot, Drug News Perspect, 2010, 23(6):341-50 [Non-Patent Document 5] Chernikov et. al., Front. Pharmacol., 2019, doi.org / 10.3389 / fphar.2019.00444 Summary of the Invention [Problem to be solved by the invention]

[0006] The present disclosure provides siNA molecules that target HSD17B13 to reduce or inhibit the production of hydroxysteroid dehydrogenase, and the siNA molecules contain optimized combinations and numbers of modified nucleotides, nucleotide lengths, designs (e.g., blunt ends or overhangs, internucleoside linkages, conjugates), and modification patterns that exhibit improved delivery and stability. [Means for solving the problem]

[0007] One aspect of the present disclosure is a sense strand comprising a nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638, and / or a sense strand comprising a nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 60 and an antisense strand comprising a nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% identical to the nucleotide sequence of any one of 4-637 or 639-644, wherein the siNA molecule downregulates the expression of the hydroxysteroid 17-beta dehydrogenase 13 (HSD17B13) gene.

[0008] Another aspect of the present disclosure relates to a double-stranded short interfering nucleic acid (siNA) molecule comprising a sense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603 or 638, and / or an antisense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637 or 639-644, wherein the siNA molecule downregulates expression of the hydroxysteroid 17-beta dehydrogenase 13 (HSD17B13) gene.

[0009] Another aspect of the present disclosure relates to a double-stranded short interfering nucleic acid (siNA) molecule selected from any one of siNA duplexes ID Nos. D1-D178 or MD1-MD178.

[0010] Another aspect of the present disclosure relates to a pharmaceutical composition comprising any of the siNA molecules according to the present disclosure and a pharmaceutically acceptable carrier.

[0011] Another aspect of the present disclosure relates to a method for treating an HSD17B13-related disease in a subject in need thereof, comprising administering to the subject an amount of any of the siNA molecules or pharmaceutical compositions of the present disclosure, thereby treating the subject. For example, the liver disease may be NAFLD, hepatocellular carcinoma (HCC) and / or NASH and / or fatty liver.

[0012] Another aspect of the present disclosure relates to a method for treating liver disease in a subject in need thereof, comprising administering to the subject an amount of any of the siNA molecules or pharmaceutical compositions of the present disclosure, thereby treating the subject. For example, the liver disease can be NAFLD, HCC, and / or NASH, and / or fatty liver.

[0013] Another aspect of the present disclosure relates to a method for treating liver disease in a subject in need of such treatment, wherein the method comprises administering to the subject a specific amount of either a siNA molecule or pharmaceutical composition according to the present disclosure, and further comprises administering to the subject at least one additional active agent, thereby treating the subject, wherein the at least one additional active agent is a liver disease therapeutic agent.

[0014] Another aspect of the present disclosure relates to a method for reducing the expression level of HSD17B13 in a patient in need thereof, the method comprising administering to the patient a specific amount of either an siRNA molecule or a pharmaceutical composition according to the present disclosure, thereby reducing the expression level of HSD17B13 in the patient.

[0015] The technology provides a method for preparing a sense strand comprising: (a) a sense strand comprising a first nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to an RNA corresponding to a target gene, wherein the first nucleotide sequence (i) is 15 to 30 nucleotides in length; and (ii) comprises 15 or more modified nucleotides independently selected from 2'-O-methyl nucleotides and 2'-fluoro nucleotides, wherein at least one modified nucleotide is a 2'-O-methyl nucleotide, and the nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. and (b) an antisense strand comprising a second nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% complementary to RNA corresponding to the target gene, wherein the second nucleotide sequence (i) is 15 to 30 nucleotides in length; and (ii) comprises 15 or more modified nucleotides independently selected from 2'-O-methyl nucleotides and 2'-fluoro nucleotides, wherein at least one modified nucleotide is a 2'-O-methyl nucleotide and at least one modified nucleotide is a 2'-fluoro nucleotide.

[0016] The present technology further provides a compound represented by formula (VIII) [ka] wherein the top strand is a sense strand comprising a first nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to an RNA corresponding to a target gene (wherein the first nucleotide sequence comprises 15-30 nucleotides); the bottom strand is an antisense strand comprising a second nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% complementary to an RNA corresponding to a target gene (wherein the second nucleotide sequence comprises 15-30 nucleotides); each A is independently a 2'-O-methyl nucleotide, or a nucleotide comprising a 5'-stabilizing end cap or a phosphorylation blocker; B is a 2'-fluoro nucleotide; C represents an overhanging nucleotide and is a 2'-O-methyl nucleotide, a deoxynucleotide, or uracil; 1 = 1-6 nucleotides in length; each n 2 , n 6 , n 8 , q 3 , q 5 , q 7 , q 9 , q 11 and q 12 are independently 0-1 nucleotides in length; each n 3 and n 4 are independently 1-3 nucleotides in length; n 5 is 1-10 nucleotides in length; n 7 is 0-4 nucleotides in length; each n 9 , q 1 and q 2 are independently 0-2 nucleotides in length; q 4 is 0-3 nucleotides in length; q 6 is 0-5 nucleotides in length; q 8 is 2-7 nucleotides in length; and q 10 are 2-11 nucleotides in length.

[0017] In any embodiment, the first nucleotide sequence comprises the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314, or 315, and / or the second nucleotide sequence comprises the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, or 288-313. In any embodiment, the first nucleotide sequence comprises the nucleotide sequence of any one of SEQ ID NOs: 316-445, 576-603, or 638, and / or the second nucleotide sequence comprises the nucleotide sequence of any one of SEQ ID NOs: 446-575, 604-637, or 639-644.

[0018] In some embodiments, the siNA reduces or inhibits the production of hydroxysteroid dehydrogenase. In some embodiments, the siNA reduces the expression or activity of HSD17B13.

[0019] In any embodiment, the sense strand and / or antisense strand disclosed herein may further comprise a TT sequence adjacent to the first and / or second nucleotide sequence. In any embodiment, the sense strand and / or antisense strand disclosed herein may further comprise phosphorothioate internucleoside linkage(s), mesyl phosphoramidate internucleoside linkage(s), 5'-stabilizing end cap(s), phosphorylation blocker(s), galactosamine(s), conjugate(s) disclosed herein, destabilizing nucleotide(s) disclosed herein, modified nucleotide(s) disclosed herein, thermally destabilizing nucleotide(s), or a combination of two or more thereof. In some embodiments, the 5' stabilized end cap(s), the phosphorylation blocker(s), the conjugate moiety(s) disclosed herein, the galactosamine(s), the destabilized nucleotide(s) disclosed herein, the modified nucleotide(s) disclosed herein, the thermally destabilized nucleotide(s), or a combination of two or more thereof, are attached to the sense strand and / or the antisense strand via one or more linkers independently selected from a phosphodiester linker, a phosphorothioate linker, or a phosphorodithioate linker.

[0020] Further disclosed herein are compositions and medicaments comprising any of the siNAs disclosed herein.

[0021] In any embodiment, the siNA molecules, compositions and / or medicaments disclosed herein can be used in the treatment of a disease, such as a liver disease, including non-alcoholic fatty liver disease (NAFLD), hepatocellular carcinoma (HCC), or non-alcoholic steatohepatitis (NASH). [Brief explanation of the drawings]

[0022] [Figure 1] FIG. 1 shows an exemplary siNA molecule. [Figure 2] FIG. 2 shows an exemplary siNA molecule. [Figures 3A-3H] 3A-3H show exemplary double-stranded siNA molecules. [Figure 4] Figures 4 and 5 show knockdown of HSD17B13 mRNA by modified siNA duplexes of the present disclosure at 7 days post-administration. [Figure 5] Figures 4 and 5 show knockdown of HSD17B13 mRNA by modified siNA duplexes of the present disclosure at 7 days post-administration. [Figure 6] FIG. 6 shows knockdown of HSD17B13 mRNA by modified siNA duplexes of the present disclosure at 1.5 mpk at day 7 post-dose and at day 7 post-dose. [Figure 7] FIG. 7 shows knockdown of HSD17B13 mRNA by modified siNA duplexes of the present disclosure. [Figure 8] FIG. 8 shows knockdown of HSD17B13 mRNA by modified siNA duplexes of the present disclosure. [Figure 9] FIG. 9 shows knockdown of HSD17B13 mRNA by modified siNA duplexes of the present disclosure. [Figure 10] FIG. 10 shows knockdown of HSD17B13 mRNA by modified siNA duplexes of the present disclosure. [Figure 11] FIG. 11 shows a Western blot demonstrating knockdown of HSD17B13 protein by ds-siNA 137, ds-siNA 144, ds-siNA 148, and ds-siNA 151 at 7 days post-administration. [Figure 12] FIG. 12 shows the quantification of the Western blot from FIG. [Figure 13]FIG. 13 shows knockdown of HSD17B13 mRNA by ds-siNA 137, ds-siNA 144, ds-siNA 148, and ds-siNA 151 at 7 days after administration. [Figure 14] FIG. 14 shows a Western blot demonstrating knockdown of HSD17B13 protein by ds-siNA 137, ds-siNA 144, ds-siNA 148, and ds-siNA 151 at 14 days post-administration. [Figure 15] FIG. 15 shows the quantification of the Western blot from FIG. [Figure 16] FIG. 16 shows knockdown of HSD17B13 mRNA by ds-siNA 137, ds-siNA 144, ds-siNA 148, and ds-siNA 151 at 14 days post-administration. [Figure 17] FIG. 17 shows knockdown of HSD17B13 mRNA by modified siNA duplexes of the present disclosure. [Figure 18] FIG. 18 shows knockdown of HSD17B13 mRNA by modified siNA duplexes of the present disclosure. [Figure 19] FIG. 19 shows knockdown of HSD17B13 mRNA by modified siNA duplexes of the present disclosure. [Figure 20] FIG. 20 shows knockdown of HSD17B13 mRNA by modified siNA duplexes of the present disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0023] This section provides a detailed description of many different aspects and embodiments representative of the present disclosure. This description is by way of several representative examples with varying details and specificity. Other features and advantages of these embodiments will be apparent from the additional description provided herein, including the various examples. The provided examples illustrate different components and methodologies useful in implementing various embodiments of the present disclosure. The examples are not intended to limit the claimed disclosure. Based on this disclosure, one skilled in the art will be able to identify and employ other components and methodologies useful in implementing the present disclosure.

[0024] The present disclosure will be better understood with reference to the following definitions.

[0025] definition Unless otherwise defined, all technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0026] As used herein, the terms "a" and "an" mean "one or more" and include the plural unless the context is inappropriate.

[0027] As used herein, the term "about" when referring to measurable values ​​(e.g., weight, time, and dosage) is intended to encompass variations such as ±10%, ±5%, ±1%, or ±0.1% of the specified value.

[0028] Unless otherwise indicated, all numerical values ​​expressing quantities of ingredients, reaction conditions, and the like used in the specification and claims should be understood to be modified in all instances by the term "about," whether or not the term "about" precedes the numerical value. Accordingly, unless otherwise indicated, the numerical parameters set forth in the following specification and appended claims are approximations that may vary depending upon the desired properties sought to be obtained by the present disclosure. At the very least, each numerical parameter should be construed in light of the number of significant digits and ordinary rounding practices, so as not to be viewed as an attempt to limit the application of the doctrine of equivalents to the scope of the claims.

[0029] Furthermore, the disclosure of a numerical range within this specification is considered to be a disclosure of all numerical values ​​and ranges within that range. For example, if a range is from about 1 to about 50, it is considered to encompass, for example, 1, 50, 7, 34, 46.1, 23.7, or any other value or range within that range. Furthermore, as used herein, the term "at least" is inclusive of the recited number, for example, "at least 50" includes 50.

[0030] As a general matter, compositions specifying percentages specify percent by weight unless otherwise specified. Further, if a variable is not accompanied by a definition, the previous definition for that variable is followed.

[0031] The term "including" is used herein to mean, and is used interchangeably with, the phrase "including, but not limited to."

[0032] As used herein, the terms "siRNA" and "siRNA molecule" and "siNA" and "siNA molecule" are used interchangeably and refer to short (or small) interfering ribonucleic acid (RNA), including chemically modified RNA, which can be single-stranded or double-stranded. As used herein, siRNA can include modified nucleotides (which include modifications in the sugar, nucleobase, and / or phosphodiester backbone (internucleoside linkage)) and nucleoside analogs, as well as conjugates or ligands. As used herein, the term "siNA duplex" or "siRNA duplex" refers to a double-stranded ("ds") siRNA or "dsRNA" or "ds-NA" having a sense strand and an antisense strand.

[0033] As used herein, the term "backbone" refers to the polymeric sugar backbone of naturally occurring nucleic acids, as well as modified counterparts and mimetics to which the nucleobases that define the base sequence of a particular nucleic acid molecule are covalently attached. In some embodiments, the backbone comprises phosphodiester internucleoside linkages (in which case it is referred to as a "phosphodiester backbone"). In some embodiments, in addition to phosphodiester internucleoside linkages, the backbone comprises one or more non-phosphodiester internucleoside linkages (e.g., phosphorothioate internucleoside linkages), as described herein. In some embodiments, a phosphodiester internucleoside linkage connects the 3'-position of the sugar moiety (e.g., ribose) of a preceding nucleoside to the 5'-position of the sugar moiety of a succeeding nucleoside (3'-5' phosphodiester linkage). In some embodiments, a phosphodiester internucleoside linkage links the 2' position of the sugar moiety (e.g., ribose) of a preceding nucleoside to the 5' position of the sugar moiety of a succeeding nucleoside (2'-5' phosphodiester linkage). Similarly, a non-phosphodiester internucleoside linkage (e.g., phosphorothioate internucleoside linkage) can link the 3' position of the sugar moiety (e.g., ribose) of a preceding nucleoside to the 5' position of the sugar moiety of a succeeding nucleoside (3'-5' phosphorothioate linkage), or can link the 2' position of the sugar moiety (e.g., ribose) of a preceding nucleoside to the 5' position of the sugar moiety of a succeeding nucleoside (2'-5' phosphorothioate linkage). In some embodiments, the siRNA exclusively comprises 3'-5' internucleoside linkages. In some embodiments, the siRNA comprises exclusively 2'-5' internucleoside linkages. In some embodiments, the siRNA comprises a mixture of 3'-5' and 2'-5' internucleoside linkages.

[0034] As used herein, the term "antisense strand" or "guide strand" refers to the strand of an siRNA molecule that includes a region that is substantially complementary to a target sequence (eg, HSD17B13 mRNA).

[0035] As used herein, the term "sense strand" or "passenger strand" refers to the strand of an siRNA molecule that includes a region that is substantially complementary to a region of the antisense strand, as defined herein.

[0036] As used herein, the term "modified nucleotide" refers to a nucleotide having a modification independently in the sugar, nucleobase, and / or phosphodiester backbone (internucleoside linkage), as well as a nucleoside analog. Thus, the term "modified nucleotide" encompasses the substitution, addition, or removal of, for example, a functional group or atom, to the internucleoside linkage, sugar moiety, or nucleobase. Modifications suitable for use in the siRNA of the present disclosure include all types of modifications disclosed herein or known in the art. Any such modifications used in siNA molecules or siRNA molecules are encompassed by "siRNA" and "siRNA molecule" and "siRNA duplex" and "siNA" and "siNA molecule" and "siNA duplex" for the purposes of this specification and claims. It will be understood that the term "nucleotide" can also refer to a modified nucleotide, as further detailed herein.

[0037] As used herein, the term "nucleobase" refers to naturally occurring nucleobases and analogs thereof. Examples of naturally occurring nucleobases or analogs thereof include, but are not limited to, thymine, uracil, adenine, cytosine, guanine, aryl, heteroaryl, and analogs or derivatives thereof.

[0038] As used herein, the term "nucleotide overhang" or "overhang" refers to at least one unpaired nucleotide extending beyond the duplex structure of a double-stranded RNA (e.g., an siRNA duplex or a dsRNA). For example, a nucleotide overhang exists when the 3' end of one strand of a dsRNA extends beyond the 5' end of the other strand, or vice versa. The overhang may be present on the sense strand, the antisense strand, or a combination thereof. Furthermore, the overhanging nucleotides may be present at the 5' end, the 3' end, or both ends of the antisense and / or sense strands of the dsRNA and may comprise modified nucleotides. Generally, when a nucleotide overhang, as defined herein, is present, the sequence of such an overhang is not considered when determining the degree of complementarity between two sequences, and such an overhang is not considered a mismatch in determining complementarity. By way of example, if a sense strand 21 nucleotides in length and an antisense strand 21 nucleotides in length hybridize to form a 19 base pair duplex region with a 2 nucleotide overhang at the 3' end of each strand, the sense and antisense strands are considered to be fully complementary, as that term is used herein.

[0039] As used herein, the term "blunt end" refers to an end of a dsRNA that has no unpaired nucleotides, i.e., no overhanging nucleotides. In some embodiments, the blunt end can be present at one or both ends of the dsRNA.

[0040] As used herein, the terms "complementary," "fully complementary," and "substantially complementary" can be used to refer to base pairing between the sense and antisense strands of a double-stranded siRNA or dsRNA, or between the antisense strand of an siRNA and a target sequence, as understood from the context of their use. As used herein, a first sequence is "complementary" to a second sequence if a polynucleotide comprising the first sequence can hybridize to a polynucleotide comprising the second sequence under specific conditions, such as physiological conditions, to form a duplex region. Other such conditions include moderate or stringent hybridization conditions known to those skilled in the art. A first sequence is considered to be fully complementary (100% complementary) to a second sequence if the polynucleotide comprising the first sequence forms base pairs with the polynucleotide comprising the second sequence without mismatches along the entire length of one or both nucleotide sequences. In some embodiments, a particular sequence is "substantially complementary" to a target sequence if it is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to the target sequence. The percentage of complementarity can be calculated, for example, by dividing the number of bases in a first sequence that are complementary to bases at corresponding positions in a second sequence or target sequence by the total length of the first sequence. Such calculations are within the capabilities of those skilled in the art. Furthermore, a particular sequence can be said to be substantially complementary to another sequence if, when the two sequences are hybridized, there are no more than five, more than four, more than three, more than two, or more than one mismatch over a duplex region of, for example, 30 base pairs. "Complementary" sequences, as used herein, may also include, or be formed entirely from, non-Watson-Crick base pairs and / or base pairs formed from modified nucleotides, so long as the above requirements regarding the ability to hybridize are met.

[0041] The use of percent identity (i.e., "identical") is a general method for defining the number of nucleobase differences between two nucleic acid sequences. For example, if the first sequence is ACGT, the second sequence of ACGA will be considered a "non-identical" sequence with one difference. Percent identity can be calculated over the entire length of the sequence, or over a portion of the sequence. Percent identity can be calculated according to the number of nucleobases that have identical base pairs corresponding to the sequences being compared. Non-identical nucleobases can be adjacent to each other, can be dispersed throughout the sequence, or both. Such calculations are within the capabilities of those skilled in the art.

[0042] As used herein, a "missense mutation" refers to a case where a single base pair change results in the substitution of a different amino acid in the resulting protein.

[0043] As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount of an siRNA of the present disclosure sufficient to produce a beneficial or desired result, e.g., an amount that elicits a biological or medical response in a tissue, system, animal, or human that is sought by a researcher, veterinarian, physician, or other clinician. A therapeutically effective amount can be administered in one or more administrations, applications, or doses, and is not intended to be limited to a particular formulation or route of administration. In some embodiments, a "therapeutically effective amount" refers to an amount that alleviates at least one clinical symptom in a human patient (e.g., at least one symptom of an HSD17B13-associated disease or liver disease).

[0044] As used herein, the terms "patient" and "subject" refer to an organism that uses the siRNA molecules of the present disclosure for the prevention or treatment of a medical condition (including the prevention or treatment in the methods of the present disclosure). Such organisms are preferably mammals, and more preferably humans. As used herein, a subject "in need" of treatment or prophylactic treatment of an existing condition encompasses both a determination of need by a medical professional and the patient's desire for such treatment. Administration of a compound (e.g., a siNA or siRNA of the present disclosure) to the subject encompasses both self-administration and administration to the patient by another.

[0045] As used herein, the terms "active agent" or "active ingredient" or "therapeutic agent" refer to a component that has a pharmacological effect, such as a therapeutic effect, at an appropriate dose. This includes siRNA molecules according to the present disclosure.

[0046] As used herein, a "liver disease therapeutic agent" is an active agent that can be used to treat liver disease, alone or in combination with another active agent, other than an siRNA of the present disclosure.

[0047] As used herein, the term "pharmaceutical composition" refers to a combination of at least one active agent with an inert or active carrier, which carrier makes the composition particularly suitable for in vivo or ex vivo diagnostic or therapeutic use. In some embodiments, the term "pharmaceutical composition" refers to a composition comprising the siRNA molecules described herein and at least one additional component, which is selected from pharmaceutically acceptable carriers, diluents, adjuvants, excipients, or vehicles, such as preservatives, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavorings, perfumes, antibacterial agents, antifungal agents, lubricants, and dispensing agents, depending on the mode and dosage form used.

[0048] As used herein, the term "pharmaceutically acceptable carrier" refers to any pharmaceutical carrier, diluent, adjuvant, excipient, or vehicle, including those described herein, such as solvents, buffers, solutions (e.g., phosphate-buffered saline), water, emulsions (e.g., oil / water or water / oil emulsions), various types of wetting agents, stabilizers, preservatives, antibacterial and antifungal agents, dispersion media, coatings, isotonic and absorption delaying agents, etc., that are acceptable for use in formulating a medicament (e.g., a medicament suitable for administration to humans). For examples of carriers, see, for example, Martin, Remington's Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, PA

[1975] .

[0049] As used herein, the terms "treat," "treating," and "treatment" include any effect, e.g., alleviation, reduction, modulation, amelioration, or elimination, that results in the improvement of a condition, disease, disorder, or the like; or amelioration of one or more symptoms associated with a condition, disease, or disorder; or amelioration of the cause of a condition, disease, or disorder. For example, with respect to HSD17B13-associated diseases, the terms "treat," "treating," and "treatment" include, but are not limited to, alleviating or ameliorating one or more symptoms associated with HSD17B13 gene expression and / or HSD17B13 protein production (e.g., fatty liver (steatosis)), non-alcoholic steatohepatitis (NASH), cirrhosis, accumulation of fat in the liver, liver inflammation, hepatocellular necrosis, liver fibrosis, obesity, hepatocellular carcinoma (HCC), or non-alcoholic fatty liver disease (NAFLD)). "Treatment" can also mean prolonging survival as compared to expected survival in the absence of treatment.

[0050] As used herein, the terms "alleviate" and "alleviating" refer to reducing the severity of a condition and / or its symptoms, e.g., reducing the severity by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 95%.

[0051] As used herein, the terms "downregulate" or "downregulating" are used interchangeably with "reduce," "inhibit," or "suppress," or other similar terms, and encompass any level of downregulation.

[0052] As used herein, the term "HSD17B13 gene" refers to the hydroxysteroid 17-beta dehydrogenase 13 gene and includes variants thereof. HSD17B13 has the nucleotide sequence set forth in SEQ ID NO:261, which corresponds to the nucleotide sequence of the coding sequence of GenBank Accession No. NM_178135.5 (nucleotides 42-944), which is incorporated by reference in its entirety. Additional examples of HSD17B13 gene sequences, including those of other mammalian genes, are readily available using public databases (e.g., NCBI RefSeq, GenBank, UniProt, and OMIM).

[0053] Throughout this specification, when compositions are described as having, including, or comprising particular ingredients, or when processes and methods are described as having, including, or comprising particular steps, it is further intended that compositions of the disclosure exist that consist essentially of or consist of the listed ingredients, and that processes and methods of the disclosure exist that consist essentially of or consist of the listed process steps.

[0054] Unless otherwise defined, all technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this disclosure belongs. The following references provide those skilled in the art with general definitions of many of the terms used in this invention: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et al., (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). As used herein, the following terms have the following meanings unless otherwise specified. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the disclosure.

[0055] siRNA molecule Disclosed herein are double-stranded short (or small) interfering RNA (siRNA) molecules that specifically downregulate the expression of the hydroxysteroid 17-beta dehydrogenase 13 (HDS17B13) gene.

[0056] In some embodiments, the double-stranded siRNA molecule comprises: (a) a sense strand comprising a nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638; and / or (b) an antisense strand comprising a nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644.

[0057] In some embodiments, the double-stranded siRNA molecule comprises: (a) a sense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638; and / or (b) an antisense strand consisting of the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644.

[0058] In some embodiments, the double-stranded siRNA molecule comprises a sense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 1-100 or 201-230. In some embodiments, the siRNA molecule comprises an antisense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 101-200 or 231-260. In some embodiments, the siRNA molecule comprises (a) a sense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 1-100 or 201-230, and (b) an antisense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 101-200 or 231-260.

[0059] In some embodiments, the double-stranded siRNA molecule comprises a sense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 316-445. In some embodiments, the siRNA molecule comprises an antisense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 446-575. In some embodiments, the siRNA molecule comprises (a) a sense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 316-445, and (b) an antisense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 446-575.

[0060] In some embodiments, the double-stranded siRNA molecule comprises a sense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 262-287, 314, or 315. In some embodiments, the siRNA molecule comprises an antisense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 288-313. In some embodiments, the siRNA molecule comprises (a) a sense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 262-287, 314, or 315, and (b) an antisense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 288-313.

[0061] In some embodiments, the double-stranded siRNA molecule comprises a sense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 576-603 or 638. In some embodiments, the siRNA molecule comprises an antisense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 604-637 or 639-644. In some embodiments, the siRNA molecule comprises (a) a sense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 576-603 or 638, and (b) an antisense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 604-637 or 639-644.

[0062] In some embodiments, the double-stranded siRNA molecule comprises: (a) a sense strand comprising at least about 15, 16, 17, 18, 19, 20, or 21 consecutive nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638; and / or (b) an antisense strand comprising at least about 15, 16, 17, 18, 19, 20, 21, 22, or 23 consecutive nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644.

[0063] In some embodiments, the double-stranded siRNA molecule comprises: (a) a sense strand comprising a nucleotide sequence having at least about 15, 16, 17, 18, 19, 20, or 21 consecutive nucleotides of any one of the nucleotide sequences of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638; and / or (b) an antisense strand comprising a nucleotide sequence having at least about 15, 16, 17, 18, 19, 20, 21, 22, or 23 consecutive nucleotides of any one of the nucleotide sequences of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644.

[0064] In some embodiments, at least one end of the double-stranded siRNA molecule is blunt-ended.In some embodiments, both ends of the double-stranded siRNA molecule are blunt-ended.In some embodiments, one end of the double-stranded siRNA molecule comprises a blunt end, and one end of the double-stranded siRNA molecule comprises an overhang.

[0065] In some embodiments, at least one end of the siRNA molecule comprises an overhang, wherein the overhang comprises at least one unpaired nucleotide. In some embodiments, at least one end of the siRNA molecule comprises an overhang, wherein the overhang comprises at least two unpaired nucleotides. In some embodiments, both ends of the siRNA molecule comprise an overhang, wherein the overhang comprises at least one unpaired nucleotide. In some embodiments, both ends of the siRNA molecule comprise an overhang, wherein the overhang comprises at least two unpaired nucleotides. In some embodiments, the siRNA molecule comprises an overhang of two unpaired nucleotides at the 3'-end of the sense strand. In some embodiments, the siRNA molecule comprises an overhang of two unpaired nucleotides at the 3'-end of the antisense strand. In some embodiments, the siRNA molecule comprises an overhang of two unpaired nucleotides at the 3'-end of the sense strand and the 3'-end of the antisense strand.

[0066] In some embodiments, the double-stranded siRNA molecule is selected from any one of siNA duplex ID numbers ds-siNA D1-D178 or mds-siNA MD1-MD178. In some embodiments, the double-stranded siRNA molecule is selected from any one of siRNA duplex ID numbers ds-siNA D1-D178. In some embodiments, the double-stranded siRNA molecule is selected from any one of siRNA duplex ID numbers mds-siNA MD1-MD178.

[0067] In some embodiments, the double-stranded siRNA molecule is selected from any one of the siRNA duplexes in Table 8 or Table 9 or Table 10 or Table 11 or Table 12. In some embodiments, the double-stranded siRNA molecule is selected from any one of the siRNA duplexes in Table 8. In some embodiments, the double-stranded siRNA molecule is selected from any one of the siRNA duplexes in Table 9. In some embodiments, the double-stranded siRNA molecule is selected from any one of the siRNA duplexes in Table 10. In some embodiments, the double-stranded siRNA molecule is selected from any one of the siRNA duplexes in Table 11. In some embodiments, the double-stranded siRNA molecule is selected from any one of the siRNA duplexes in Table 12.

[0068] In some embodiments, the double-stranded siRNA molecule is about 17 to about 29 base pairs, or 19-23 base pairs, or 19-21 base pairs in length, one strand of which is complementary to a target mRNA and causes degradation of the target mRNA when added to a cell harboring the target mRNA or when produced in vivo within the cell.

[0069] In some embodiments, the siRNA molecule of the present disclosure comprises a nucleotide sequence complementary to the nucleotide sequence of the target gene. In some embodiments, the siRNA molecule of the present disclosure interacts with the nucleotide sequence of the target gene to cause inhibition of the expression of the target gene.

[0070] The siRNA molecule can be obtained using any one of many techniques known to those skilled in the art. In some embodiments, the siRNA molecule can be synthesized as two separate complementary nucleic acid molecules or as a single nucleic acid molecule having two complementary regions. For example, the siRNA of the present disclosure can be chemically synthesized using appropriately protected ribonucleoside phosphoramidites and a conventional RNA synthesizer or other well-known methods. Furthermore, the siRNA can be produced by commercial suppliers (e.g., Dharmacon / Horizon (Lafayette, Colo., USA), Glen Research (Sterling, Va., USA), ChemGenes (Ashland, Mass., USA), and Cruachem (Glasgow, UK)). In some embodiments, the siRNA molecule can be encoded by a plasmid.

[0071] Sense strand Any of the siRNA molecules described herein can comprise a sense strand. In some embodiments, the sense strand comprises about 15 to about 50 nucleotides. In some embodiments, the sense strand comprises about 15 to about 45 nucleotides. In some embodiments, the sense strand comprises about 15 to about 40 nucleotides. In some embodiments, the sense strand comprises about 15 to about 35 nucleotides. In some embodiments, the sense strand comprises about 15 to about 30 nucleotides. In some embodiments, the sense strand comprises about 15 to about 25 nucleotides. In some embodiments, the sense strand comprises about 17 to about 23 nucleotides. In some embodiments, the sense strand comprises about 17 to about 22 nucleotides. In some embodiments, the sense strand comprises about 17 to about 21 nucleotides. In some embodiments, the sense strand comprises about 18 to about 23 nucleotides. In some embodiments, the sense strand comprises about 18 to about 22 nucleotides. In some embodiments, the sense strand comprises about 18 to about 21 nucleotides. In some embodiments, the sense strand comprises about 19 to about 23 nucleotides. In some embodiments, the sense strand comprises about 19 to about 22 nucleotides. In some embodiments, the sense strand comprises about 19 to about 21 nucleotides.

[0072] In some embodiments, the sense strand comprises at least about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more nucleotides. In some embodiments, the sense strand comprises at least about 15 nucleotides. In some embodiments, the sense strand comprises at least about 16 nucleotides. In some embodiments, the sense strand comprises at least about 17 nucleotides. In some embodiments, the sense strand comprises at least about 18 nucleotides. In some embodiments, the sense strand comprises at least about 19 nucleotides. In some embodiments, the sense strand comprises at least about 20 nucleotides. In some embodiments, the sense strand comprises at least about 21 nucleotides. In some embodiments, the sense strand comprises at least about 22 nucleotides. In some embodiments, the sense strand comprises at least about 23 nucleotides.

[0073] In some embodiments, the sense strand contains no more than about 50, 45, 40, 35, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, or 19 nucleotides. In some embodiments, the sense strand contains fewer than about 30 nucleotides. In some embodiments, the sense strand contains fewer than about 25 nucleotides. In some embodiments, the sense strand contains fewer than about 24 nucleotides. In some embodiments, the sense strand contains fewer than about 23 nucleotides. In some embodiments, the sense strand contains fewer than about 22 nucleotides. In some embodiments, the sense strand contains fewer than about 21 nucleotides. In some embodiments, the sense strand contains fewer than about 20 nucleotides. In some embodiments, the sense strand contains fewer than about 19 nucleotides.

[0074] In some embodiments, the sense strand comprises a sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to a fragment of the HSD17B13 gene over the entire length of the sense strand. In some embodiments, the sense strand comprises a sequence that is at least about 70% identical to a fragment of the HSD17B13 gene over the entire length of the sense strand. In some embodiments, the sense strand comprises a sequence that is at least about 75% identical to a fragment of the HSD17B13 gene over the entire length of the sense strand. In some embodiments, the sense strand comprises a sequence that is at least about 80% identical to a fragment of the HSD17B13 gene over the entire length of the sense strand. In some embodiments, the sense strand comprises a sequence that is at least about 85% identical to a fragment of the HSD17B13 gene over the entire length of the sense strand. In some embodiments, the sense strand comprises a sequence that is at least about 90% identical to a fragment of the HSD17B13 gene over the entire length of the sense strand. In some embodiments, the sense strand comprises a sequence that is at least about 95% identical to a fragment of the HSD17B13 gene over the entire length of the sense strand. In some embodiments, the sense strand comprises a sequence that is about 100% identical to a fragment of the HSD17B13 gene over the entire length of the sense strand. In some embodiments, the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 15 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 16 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 17 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 18 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 19 contiguous nucleotides of the HSD17B13 gene.In some embodiments, the fragment of the HSD17B13 gene consists of about 20 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 21 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 22 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 23 contiguous nucleotides of the HSD17B13 gene.

[0075] In some embodiments, the sense strand comprises a sequence of about 15 to about 50 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence of about 15 to about 45 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence of about 15 to about 40 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence of about 15 to about 35 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence of about 15 to about 30 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence of about 15 to about 25 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises about 17 to about 23 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises about 17 to about 22 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises about 17 to about 21 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises about 18 to about 23 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises about 18 to about 22 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises about 18 to about 21 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises about 19 to about 23 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises about 19 to about 22 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises about 19 to about 21 contiguous nucleotides of a fragment of the HSD17B13 gene.In some embodiments, the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 15 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 16 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 17 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 18 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 19 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 20 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 21 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 22 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 23 contiguous nucleotides of the HSD17B13 gene.

[0076] In some embodiments, the sense strand comprises a sequence having at least about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having at least about 15 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having at least about 16 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having at least about 17 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having at least about 18 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having at least about 19 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence of at least about 20 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence of at least about 21 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence of at least about 22 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence of at least about 23 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 15 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 16 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 17 contiguous nucleotides of the HSD17B13 gene.In some embodiments, the fragment of the HSD17B13 gene consists of about 18 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 19 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 20 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 21 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 22 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 23 contiguous nucleotides of the HSD17B13 gene.

[0077] In some embodiments, the sense strand comprises a sequence having no more than about 50, 45, 40, 35, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, or 19 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having fewer than about 35 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having fewer than about 30 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having fewer than about 25 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having fewer than about 24 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having fewer than about 23 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having fewer than about 22 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having fewer than about 21 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having fewer than about 20 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having fewer than about 19 contiguous nucleotides of a fragment of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 15 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 16 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 17 contiguous nucleotides of the HSD17B13 gene.In some embodiments, the fragment of the HSD17B13 gene consists of about 18 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 19 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 20 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 21 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 22 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 23 contiguous nucleotides of the HSD17B13 gene.

[0078] In some embodiments, the sense strand comprises a sequence having no more than 5, 4, 3, 2, or 1 nucleobase difference from a fragment of the HSD17B13 gene over the entire length of the sense strand, wherein the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having no more than 5 nucleobase difference from a fragment of the HSD17B13 gene over the entire length of the sense strand, wherein the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having no more than 4 nucleobase differences from a fragment of the HSD17B13 gene over the entire length of the sense strand, wherein the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having no more than 3 nucleobase differences from a fragment of the HSD17B13 gene over the entire length of the sense strand, wherein the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having no more than two nucleobase differences from a fragment of the HSD17B13 gene over the entire length of the sense strand, wherein the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 consecutive nucleotides of the HSD17B13 gene.In some embodiments, the sense strand comprises a sequence having no more than 1 nucleobase difference from a fragment of the HSD17B13 gene along the entire length of the sense strand, wherein the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the sense strand comprises a sequence having 0 nucleobase difference from a fragment of the HSD17B13 gene along the entire length of the sense strand, wherein the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 15 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 16 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 17 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 18 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 19 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 20 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 21 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 22 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 23 contiguous nucleotides of the HSD17B13 gene.

[0079] In some embodiments, the sense strand comprises the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the sense strand comprises a nucleotide sequence over the entire length of the sense strand that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the sense strand comprises a nucleotide sequence over the entire length of the sense strand that is at least about 70% identical to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the sense strand comprises a nucleotide sequence over the entire length of the sense strand that is at least about 75% identical to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the sense strand comprises a nucleotide sequence over the entire length of the sense strand that is at least about 80% identical to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the sense strand comprises a nucleotide sequence over the entire length of the sense strand that is at least about 85% identical to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the sense strand comprises a nucleotide sequence over the entire length of the sense strand that is at least about 90% identical to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the sense strand comprises a nucleotide sequence over the entire length of the sense strand that is at least about 95% identical to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638.In some embodiments, the sense strand comprises a nucleotide sequence that is about 100% identical to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638 over the entire length of the sense strand.

[0080] In some embodiments, the sense strand comprises at least about 15, 16, 17, 18, 19, 20, or 21 contiguous nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the sense strand comprises at least about 17 contiguous nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the sense strand comprises at least about 18 contiguous nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the sense strand comprises at least about 19 contiguous nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the sense strand comprises at least about 20 contiguous nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the sense strand comprises at least about 21 contiguous nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638.

[0081] In some embodiments, the sense strand comprises a nucleotide sequence that has no more than 5, 4, 3, 2, or 1 mismatches to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644 over the entire length of the sense strand. In some embodiments, the sense strand comprises a nucleotide sequence that has no more than 5 mismatches to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644 over the entire length of the sense strand. In some embodiments, the sense strand comprises a nucleotide sequence that has no more than 4 mismatches to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644 over the entire length of the sense strand. In some embodiments, the sense strand comprises a nucleotide sequence that has three or fewer mismatches to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644 over the entire length of the sense strand. In some embodiments, the sense strand comprises a nucleotide sequence that has two or fewer mismatches to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644 over the entire length of the sense strand. In some embodiments, the sense strand comprises a nucleotide sequence that has one or fewer mismatches to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644 over the entire length of the sense strand. In some embodiments, the sense strand comprises a nucleotide sequence that has zero mismatches to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644 over the entire length of the sense strand.

[0082] In some embodiments, the sense strand comprises the nucleotide sequence of any of the sense strands listed in Table 8 or Table 9 or Table 10 or Table 11 or Table 12. In some embodiments, the sense strand comprises the nucleotide sequence of any of the sense strands listed in Table 8. In some embodiments, the sense strand comprises the nucleotide sequence of any of the sense strands listed in Table 9. In some embodiments, the sense strand comprises the nucleotide sequence of any of the sense strands listed in Table 10. In some embodiments, the sense strand comprises the nucleotide sequence of any of the sense strands listed in Table 11. In some embodiments, the sense strand comprises the nucleotide sequence of any of the sense strands listed in Table 12.

[0083] In some embodiments, the sense strand may comprise an overhang sequence. In some embodiments, the overhang sequence comprises at least about 1, 2, 3, 4, or 5 or more nucleotides. In some embodiments, the overhang sequence comprises at least about 1 nucleotide. In some embodiments, the overhang sequence comprises at least about 2 nucleotides. In some embodiments, the overhang sequence comprises at least about 3 nucleotides. In some embodiments, the overhang sequence comprises at least about 4 nucleotides. In some embodiments, the overhang sequence comprises at least about 5 nucleotides.

[0084] In some embodiments, the sense strand may comprise at least 1, 2, 3, or 4 phosphorothioate internucleoside linkages. In some embodiments, at least one phosphorothioate internucleoside linkage is present between the 1st and 2nd nucleotides from the 5'-end of the sense strand. In some embodiments, at least one phosphorothioate internucleoside linkage is present between the 2nd and 3rd nucleotides from the 5'-end of the sense strand. In some embodiments, at least one phosphorothioate internucleoside linkage is present between the 1st and 2nd nucleotides from the 3'-end of the sense strand. In some embodiments, at least one phosphorothioate internucleoside linkage is present between the 2nd and 3rd nucleotides from the 3'-end of the sense strand.

[0085] In some embodiments, the sense strand may comprise a nucleotide sequence comprising 2'-fluoro nucleotides at positions 3, 7-9, 12, and 17. In some embodiments, the sense strand may comprise a nucleotide sequence comprising 2'-fluoro nucleotides at positions 3, 7, 8, and 17. In some embodiments, the sense strand may comprise a nucleotide sequence comprising 2'-fluoro nucleotides at positions 5 and 7-9 from the 5' end of the nucleotide sequence. In some embodiments, the sense strand may comprise a nucleotide sequence comprising 2'-fluoro nucleotides at positions 7 and 9-11 from the 5' end of the nucleotide sequence. In some embodiments, the sense strand may comprise a nucleotide sequence comprising 2'-fluoro nucleotides at positions 5, 9-11, 14, and 19 from the 5' end of the nucleotide sequence. In some embodiments, the sense strand may comprise a nucleotide sequence consisting of 19-23 or 19-21 nucleotides, wherein 2'-fluoro nucleotides are present at positions 5 and 7-9 from the 5' end of the nucleotide sequence. In some embodiments, the sense strand can comprise a nucleotide sequence of 19-23 or 19-21 nucleotides, wherein 2'-fluoro nucleotides are present at positions 7 and 9-11 from the 5' end of the nucleotide sequence. In some embodiments, the sense strand can comprise a nucleotide sequence of 19-23 or 19-21 nucleotides, wherein 2'-fluoro nucleotides are present at positions 5, 9-11, 14, and 19 from the 5' end of the nucleotide sequence. In some embodiments, the nucleotides at positions 5, 9, 10, and / or 11 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides.

[0086] antisense strand Any of the siRNA molecules described herein can comprise an antisense strand. In some embodiments, the antisense strand comprises about 15 to about 50 nucleotides. In some embodiments, the antisense strand comprises about 15 to about 45 nucleotides. In some embodiments, the antisense strand comprises about 15 to about 40 nucleotides. In some embodiments, the antisense strand comprises about 15 to about 35 nucleotides. In some embodiments, the antisense strand comprises about 15 to about 30 nucleotides. In some embodiments, the antisense strand comprises about 15 to about 25 nucleotides. In some embodiments, the antisense strand comprises about 17 to about 23 nucleotides. In some embodiments, the antisense strand comprises about 17 to about 22 nucleotides. In some embodiments, the antisense strand comprises about 17 to about 21 nucleotides. In some embodiments, the antisense strand comprises about 18 to about 23 nucleotides. In some embodiments, the antisense strand comprises about 18 to about 22 nucleotides. In some embodiments, the antisense strand comprises about 18 to about 21 nucleotides. In some embodiments, the antisense strand comprises about 19 to about 23 nucleotides. In some embodiments, the antisense strand comprises about 19 to about 22 nucleotides. In some embodiments, the antisense strand comprises about 19 to about 21 nucleotides.

[0087] In some embodiments, the antisense strand comprises at least about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more nucleotides. In some embodiments, the antisense strand comprises at least about 15 nucleotides. In some embodiments, the antisense strand comprises at least about 16 nucleotides. In some embodiments, the antisense strand comprises at least about 17 nucleotides. In some embodiments, the antisense strand comprises at least about 18 nucleotides. In some embodiments, the antisense strand comprises at least about 19 nucleotides. In some embodiments, the antisense strand comprises at least about 20 nucleotides. In some embodiments, the antisense strand comprises at least about 21 nucleotides. In some embodiments, the antisense strand comprises at least about 22 nucleotides. In some embodiments, the antisense strand comprises at least about 23 nucleotides.

[0088] In some embodiments, the antisense strand contains fewer than about 50, 45, 40, 35, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, or 19 nucleotides. In some embodiments, the antisense strand contains fewer than about 30 nucleotides. In some embodiments, the antisense strand contains fewer than about 25 nucleotides. In some embodiments, the antisense strand contains fewer than about 24 nucleotides. In some embodiments, the antisense strand contains fewer than about 23 nucleotides. In some embodiments, the antisense strand contains fewer than about 22 nucleotides. In some embodiments, the antisense strand contains fewer than about 21 nucleotides. In some embodiments, the antisense strand contains fewer than about 20 nucleotides. In some embodiments, the antisense strand contains fewer than about 19 nucleotides.

[0089] In some embodiments, the antisense strand comprises a sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% complementary to a fragment of the HSD17B13 gene over the entire length of the antisense strand. In some embodiments, the antisense strand comprises a sequence that is at least about 70% complementary to a fragment of the HSD17B13 gene over the entire length of the antisense strand. In some embodiments, the antisense strand comprises a sequence that is at least about 75% complementary to a fragment of the HSD17B13 gene over the entire length of the antisense strand. In some embodiments, the antisense strand comprises a sequence that is at least about 80% complementary to a fragment of the HSD17B13 gene over the entire length of the antisense strand. In some embodiments, the antisense strand comprises a sequence that is at least about 85% complementary to a fragment of the HSD17B13 gene over the entire length of the antisense strand. In some embodiments, the antisense strand comprises a sequence that is at least about 90% complementary to a fragment of the HSD17B13 gene over the entire length of the antisense strand. In some embodiments, the antisense strand comprises a sequence that is at least about 95% complementary to a fragment of the HSD17B13 gene over the entire length of the antisense strand. In some embodiments, the antisense strand comprises a sequence that is about 100% complementary to a fragment of the HSD17B13 gene over the entire length of the antisense strand. In some embodiments, the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 consecutive nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 15 consecutive nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 16 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 17 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 18 contiguous nucleotides of the HSD17B13 gene.In some embodiments, the fragment of the HSD17B13 gene consists of about 19 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 20 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 21 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 22 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 23 contiguous nucleotides of the HSD17B13 gene.

[0090] In some embodiments, the antisense strand comprises a sequence having about 15 to about 50 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having about 15 to about 45 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having about 15 to about 40 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having about 15 to about 35 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having about 15 to about 30 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having about 15 to about 25 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises about 17 to about 23 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises about 17 to about 22 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises about 17 to about 21 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises about 18 to about 23 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises about 18 to about 22 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises about 18 to about 21 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises about 19 to about 23 contiguous nucleotides complementary to a fragment of the HSD17B13 gene, hi some embodiments, the antisense strand comprises about 19 to about 22 contiguous nucleotides complementary to a fragment of the HSD17B13 gene.In some embodiments, the antisense strand comprises about 19 to about 21 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 15 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 16 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 17 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 18 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 19 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 20 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 21 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 22 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 23 contiguous nucleotides of the HSD17B13 gene.

[0091] In some embodiments, the antisense strand comprises a sequence having at least about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having at least about 15 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having at least about 16 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having at least about 17 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having at least about 18 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having at least about 19 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having at least about 20 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having at least about 21 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having at least about 22 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having at least about 23 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 15 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 16 contiguous nucleotides of the HSD17B13 gene.In some embodiments, the fragment of the HSD17B13 gene consists of about 17 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 18 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 19 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 20 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 21 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 22 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 23 contiguous nucleotides of the HSD17B13 gene.

[0092] In some embodiments, the antisense strand comprises a sequence having no more than about 50, 45, 40, 35, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, or 19 contiguous nucleotides that are complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having fewer than about 35 contiguous nucleotides that are complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having fewer than about 30 contiguous nucleotides that are complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having fewer than about 25 contiguous nucleotides that are complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having fewer than about 24 contiguous nucleotides that are complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having fewer than about 23 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having fewer than about 22 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having fewer than about 21 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having fewer than about 20 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having fewer than about 19 contiguous nucleotides complementary to a fragment of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 15 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 16 contiguous nucleotides of the HSD17B13 gene.In some embodiments, the fragment of the HSD17B13 gene consists of about 17 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 18 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 19 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 20 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 21 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 22 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 23 contiguous nucleotides of the HSD17B13 gene.

[0093] In some embodiments, the antisense strand comprises a sequence having no more than 5, 4, 3, 2, or 1 mismatch to a fragment of the HSD17B13 gene over the entire length of the antisense strand, wherein the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 consecutive nucleotides of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having no more than 5 mismatches to a fragment of the HSD17B13 gene over the entire length of the antisense strand, wherein the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 consecutive nucleotides of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having no more than 4 mismatches to a fragment of the HSD17B13 gene along the entire length of the antisense strand, wherein the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 consecutive nucleotides of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having no more than 3 mismatches to a fragment of the HSD17B13 gene along the entire length of the antisense strand, wherein the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 consecutive nucleotides of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having no more than two mismatches to a fragment of the HSD17B13 gene over the entire length of the antisense strand, wherein the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 consecutive nucleotides of the HSD17B13 gene.In some embodiments, the antisense strand comprises a sequence having no more than 1 mismatch to a fragment of the HSD17B13 gene along the entire length of the antisense strand, wherein the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 consecutive nucleotides of the HSD17B13 gene. In some embodiments, the antisense strand comprises a sequence having 0 mismatches to a fragment of the HSD17B13 gene along the entire length of the antisense strand, wherein the fragment of the HSD17B13 gene consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 consecutive nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 15 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 16 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 17 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 18 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 19 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 20 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 21 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 22 contiguous nucleotides of the HSD17B13 gene. In some embodiments, the fragment of the HSD17B13 gene consists of about 23 contiguous nucleotides of the HSD17B13 gene.

[0094] In some embodiments, the antisense strand comprises the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644. In some embodiments, the antisense strand comprises a nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644 over the entire length of the antisense strand. In some embodiments, the antisense strand comprises a nucleotide sequence that is at least about 70% identical to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644 over the entire length of the antisense strand. In some embodiments, the antisense strand comprises a nucleotide sequence that is at least about 75% identical to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644 over the entire length of the antisense strand. In some embodiments, the antisense strand comprises a nucleotide sequence that is at least about 80% identical to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644 over the entire length of the antisense strand. In some embodiments, the antisense strand comprises a nucleotide sequence that is at least about 85% identical to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644 over the entire length of the antisense strand. In some embodiments, the antisense strand comprises a nucleotide sequence that is at least about 90% identical to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644 over the entire length of the antisense strand.In some embodiments, the antisense strand comprises a nucleotide sequence that is at least about 95% identical to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644 over the entire length of the antisense strand. In some embodiments, the antisense strand comprises a nucleotide sequence that is about 100% identical to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644 over the entire length of the antisense strand.

[0095] In some embodiments, the antisense strand comprises at least about 15, 16, 17, 18, 19, 20, 21, 22, or 23 consecutive nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644. In some embodiments, the antisense strand comprises at least about 17 consecutive nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644. In some embodiments, the antisense strand comprises at least about 18 contiguous nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644. In some embodiments, the antisense strand comprises at least about 19 contiguous nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644. In some embodiments, the antisense strand comprises at least about 20 contiguous nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644. In some embodiments, the antisense strand comprises at least about 21 contiguous nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644. In some embodiments, the antisense strand comprises at least about 22 contiguous nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644. In some embodiments, the antisense strand comprises at least about 23 contiguous nucleotides of the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644.

[0096] In some embodiments, the antisense strand comprises a nucleotide sequence that has 5, 4, 3, 2, or 1 or less mismatches over the entire length of the antisense strand to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the antisense strand comprises a nucleotide sequence that has 5 or less mismatches over the entire length of the antisense strand to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the antisense strand comprises a nucleotide sequence that has 4 or less mismatches over the entire length of the antisense strand to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the antisense strand comprises a nucleotide sequence that has three or fewer mismatches over the entire length of the antisense strand to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the antisense strand comprises a nucleotide sequence that has two or fewer mismatches over the entire length of the antisense strand to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the antisense strand comprises a nucleotide sequence that has one or fewer mismatches over the entire length of the antisense strand to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638. In some embodiments, the antisense strand comprises a nucleotide sequence having zero mismatches to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638 over the entire length of the antisense strand.

[0097] In some embodiments, the antisense strand comprises the nucleotide sequence of any of the antisense strands listed in Table 8 or Table 9 or Table 10 or Table 11 or Table 12. In some embodiments, the antisense strand comprises the nucleotide sequence of any of the antisense strands listed in Table 8. In some embodiments, the antisense strand comprises the nucleotide sequence of any of the antisense strands listed in Table 9. In some embodiments, the antisense strand comprises the nucleotide sequence of any of the antisense strands listed in Table 10. In some embodiments, the antisense strand comprises the nucleotide sequence of any of the antisense strands listed in Table 11. In some embodiments, the antisense strand comprises the nucleotide sequence of any of the antisense strands listed in Table 12.

[0098] In some embodiments, the antisense strand may comprise an overhang sequence at either the 3' or 5' end. In some embodiments, the overhang sequence comprises at least about 1, 2, 3, 4, or 5 or more nucleotides. In some embodiments, the overhang sequence comprises at least about 1 nucleotide. In some embodiments, the overhang sequence comprises at least about 2 nucleotides. In some embodiments, the overhang sequence comprises at least about 3 nucleotides. In some embodiments, the overhang sequence comprises at least about 4 nucleotides. In some embodiments, the overhang sequence comprises at least about 5 nucleotides. In some embodiments, the overhang sequence comprises a UU sequence.

[0099] In some embodiments, the antisense strand may comprise at least 1, 2, 3, or 4 phosphorothioate internucleoside linkages. In some embodiments, at least one phosphorothioate internucleoside linkage is present between the 1st and 2nd nucleotides from the 5'-end of the antisense strand. In some embodiments, at least one phosphorothioate internucleoside linkage is present between the 2nd and 3rd nucleotides from the 5'-end of the antisense strand. In some embodiments, at least one phosphorothioate internucleoside linkage is present between the 1st and 2nd nucleotides from the 3'-end of the antisense strand. In some embodiments, at least one phosphorothioate internucleoside linkage is present between the 2nd and 3rd nucleotides from the 3'-end of the antisense strand.

[0100] In some embodiments, the antisense strand may comprise a nucleotide sequence comprising 2'-fluoro nucleotides at positions 2, 6, 14, and 16 from the 5' end of the nucleotide sequence. In some embodiments, the antisense strand may comprise a nucleotide sequence comprising 2'-fluoro nucleotides at positions 2 and 14 from the 5' end of the nucleotide sequence. In some embodiments, the antisense strand may comprise a nucleotide sequence comprising 2'-fluoro nucleotides at positions 2, 5, 8, 14, and 17 from the 5' end of the nucleotide sequence.

[0101] In some embodiments, the antisense strand can comprise a nucleotide sequence of 17-23 or 19-23 nucleotides, wherein 2'-fluoro nucleotides are present at positions 2, 6, 14, and 16 from the 5' end of the nucleotide sequence. In some embodiments, the antisense strand can comprise a nucleotide sequence of 17-23 or 19-23 nucleotides, wherein 2'-fluoro nucleotides are present at positions 2 and 14 from the 5' end of the nucleotide sequence. In some embodiments, the antisense strand can comprise a nucleotide sequence of 17-23 or 19-23 nucleotides, wherein 2'-fluoro nucleotides are present at positions 2, 5, 8, 14, and 17 from the 5' end of the nucleotide sequence. In some embodiments, the antisense strand can comprise a nucleotide sequence of 17-23 or 19-23 nucleotides, wherein 2'-fluoro nucleotides are present at positions 2, 6, 10, 14, and 18 from the 5' end of the nucleotide sequence.

[0102] Modified siRNA In some embodiments, the siRNA molecules disclosed herein can be chemically modified. In some embodiments, the siRNA molecules can be modified, for example, to improve stability and / or bioavailability, and / or to provide other beneficial properties in vitro, in vivo, and / or ex vivo. For example, the siRNA molecules can be modified so that the two strands (sense and antisense) maintain the ability to hybridize with each other and / or the siRNA molecules maintain the ability to hybridize with the target sequence. Examples of siRNA modifications include modifications to the ribose sugar, nucleobase, and / or phosphodiester backbone, including, but not limited to, modifications described herein. Non-limiting examples of siRNA modifications are described, for example, in: WO2020 / 243490; WO2020 / 097342; WO2021 / 119325; PCT / US2021 / 019629; PCT / US2021 / 019628; PCT / US2021 / 021199; Sig. Transduct. Target Ther.-5 (101), 1-25, 2020; and J. Am. Chem. Soc. 136 (49), 16958-16961, 2014; the contents of each of which are incorporated herein by reference in their entirety.

[0103] In some embodiments, the siRNA molecules disclosed herein comprise modified nucleotides having modifications of the ribose sugar. These sugar modifications can include modifications at the 2' and / or 5' positions of the pentose ring and bicyclic sugar modifications. A 2'-modified nucleotide refers to a nucleotide having a pentose ring with a substituent other than H or OH at the 2' position. Such 2' modifications include, but are not limited to, 2'-OH, 2'-S-alkyl, 2'-N-alkyl, 2'-O-alkyl, 2'-S-alkenyl, 2'-N-alkenyl, 2'-O-alkenyl, 2'-S-alkynyl, 2'-N-alkynyl, 2'-O-alkynyl, 2'-O-allyl, 2'-C-allyl, 2'-fluoro, 2'-O-methyl (OMe or OCH), 2'-O-methoxyethyl, 2'-ara-F, 2'-OCF, 2'-O(CH)SCH, 2'-O-aminoalkyl, 2'-amino (e.g., NH), 2'-O-ethylamine, and 2'-azido, wherein the alkyl, alkenyl, and alkynyl can be substituted or unsubstituted. Modifications at the 5' position of the pentose ring include, but are not limited to, 5'-methyl (R or S), 5'-vinyl, and 5'-methoxy. Sugar modifications can also include, for example, LNA, UNA, GNA, and DNA. In some embodiments, the siRNA molecules of the present disclosure comprise one or more 2'-O-methyl nucleotides, 2'-fluoro nucleotides, or a combination thereof.

[0104] In some embodiments, about 15-30, 15-25, 15-24, 15-23, 15-22, 15-21, 17-30, 17-25, 17-24, 17-23, 17-22, 17-21, 18-30, 18-25, 18-24, 18-23, 18-22, 18-21, 19-30, 19-25, 19-24, 19-23, 19-22, 19-21, 20-25, 20-24, 20-23, 21-25, 21-24, or 21-23 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, about 2-20 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, about 5-25 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, about 10-25 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, about 12-25 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, at least about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, at least about 12 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, at least about 13 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides.In some embodiments, at least about 14 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, at least about 15 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, at least about 16 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, at least about 17 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, at least about 18 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, at least about 19 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, no more than 25, no more than 24, no more than 23, no more than 22, no more than 21, no more than 20, no more than 19, no more than 18, no more than 17, no more than 16, no more than 15, no more than 14, no more than 13, no more than 12, no more than 11, no more than 10, no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, or no more than 2 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, no more than 21 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, no more than 20 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides.In some embodiments, no more than 19 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, no more than 18 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, no more than 17 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, no more than 16 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, no more than 15 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, no more than 14 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, no more than 13 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methyl nucleotides. In some embodiments, at least one modified nucleotide of any sense or antisense nucleotide sequence described herein is a 2'-O-methylpyrimidine. In some embodiments, at least 5, 6, 7, 8, 9, or 10 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methylpyrimidine. In some embodiments, at least one modified nucleotide of any sense or antisense nucleotide sequence described herein is a 2'-O-methylpurine. In some embodiments, at least 5, 6, 7, 8, 9, or 10 modified nucleotides of any sense or antisense nucleotide sequence described herein are 2'-O-methylpurine.In some embodiments, the 2'-O-methyl nucleotide is a 2'-O-methyl nucleotide mimic.

[0105] In some embodiments, the nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, at least two nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, at least three nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, at least four nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, at least five nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the 3rd nucleotide from the 5' end of any sense or antisense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the 7th nucleotide from the 5' end of any sense or antisense nucleotide sequence described herein is a 2'-fluoro nucleotide.In some embodiments, the nucleotide at position 8 from the 5' end of any sense or antisense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 9 from the 5' end of any sense or antisense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 12 from the 5' end of any sense or antisense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 17 from the 5' end of any sense or antisense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the 2'-fluoro nucleotide is a 2'-fluoro nucleotide mimic.

[0106] In some embodiments, at least 1, 2, 3, 4, 5, 6, or 7 nucleotides at positions 1, 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 1, 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, at least two nucleotides at positions 1, 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, at least three nucleotides at positions 1, 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 1, 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the 2'-fluoro nucleotides are 2'-fluoro nucleotide mimics.

[0107] In some embodiments, at least 1, 2, 3, 4, 5, 6, or 7 nucleotides at positions 2, 4, 6, 8, 10, 12, 14, 16, and / or 18 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 4, 6, 8, 10, 12, 14, 16, and / or 18 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, at least two nucleotides at positions 2, 4, 6, 8, 10, 12, 14, 16, and / or 18 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, at least three nucleotides at positions 2, 4, 6, 8, 10, 12, 14, 16, and / or 18 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 2, 4, 6, 8, 10, 12, 14, 16, and / or 18 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the 2'-fluoro nucleotides are 2'-fluoro nucleotide mimics.

[0108] In some embodiments, the nucleotide at position 1 from the 5' end of any sense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 3 from the 5' end of any sense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 5 from the 5' end of any sense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 7 from the 5' end of any sense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 8 from the 5' end of any sense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 9 from the 5' end of any sense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 10 from the 5' end of any sense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 11 from the 5' end of any sense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 12 from the 5' end of any sense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 14 from the 5' end of any sense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 17 from the 5' end of any sense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 19 from the 5' end of any sense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the 2'-fluoro nucleotide is a 2'-fluoro nucleotide mimic.

[0109] In some embodiments, at least 1, 2, 3, 4, 5, 6, or 7 nucleotides at positions 1, 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of any sense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 5, 7, 8, 9, 10, 11, 14, and / or 19 from the 5' end of any sense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 5, 7, 8, and / or 9 from the 5' end of any sense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 7, 9, 10, and / or 11 from the 5' end of any sense or antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the nucleotide at positions 5, 9, 10, 11, 14, and / or 19 from the 5' end of any sense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the 2'-fluoro nucleotide is a 2'-fluoro nucleotide mimic.

[0110] In some embodiments, the nucleotide at position 2 from the 5' end of any antisense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 4 from the 5' end of any antisense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 6 from the 5' end of any antisense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 8 from the 5' end of any antisense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 10 from the 5' end of any antisense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 12 from the 5' end of any antisense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 14 from the 5' end of any antisense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 16 from the 5' end of any antisense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the 18th nucleotide from the 5' end of any antisense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the 2'-fluoro nucleotide is a 2'-fluoro nucleotide mimic.

[0111] In some embodiments, at least 1, 2, 3, 4, 5, 6, or 7 nucleotides at positions 2, 4, 5, 6, 8, 10, 12, 14, 16, 17, and / or 18 from the 5' end of any antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 2, 5, 6, 8, 14, 16, and / or 17 from the 5' end of any antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 2, 6, 14, and / or 16 from the 5' end of any antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 2 and / or 14 from the 5' end of any antisense nucleotide sequence described herein are 2'-fluoro nucleotides. In some embodiments, the nucleotide at positions 2, 5, 8, 14, and / or 17 from the 5' end of any antisense nucleotide sequence described herein is a 2'-fluoro nucleotide. In some embodiments, the 2'-fluoro nucleotide is a 2'-fluoro nucleotide mimic.

[0112] In some embodiments, the 2'-fluoro nucleotide mimic or the 2'-O-methyl nucleotide mimic has formula (V): [ka] where R x are independently a nucleobase, aryl, heteroaryl, or H, and Q 1 and Q 2 are independently S or O, and R 5 are independently -OCD3, -F, or -OCH3, and R 6 and R 7 is independently H, D, or CD3. In some embodiments, the nucleobase is selected from thymine, cytosine, guanine, adenine, uracil, and analogs or derivatives thereof.

[0113] In some embodiments, the 2'-fluoro nucleotide mimic or 2'-O-methyl nucleotide mimic has Formula (16)-Formula (20): [ka] where R x are independently a nucleobase, and R 2 is F or —OCH 3 . In some embodiments, the nucleobase is selected from thymine, cytosine, guanine, adenine, uracil, and analogs or derivatives thereof.

[0114] In some embodiments, the sense strand or the antisense strand has the following chemical structure: [ka] and wherein R x is independently a nucleobase, aryl, heteroaryl, or H. In some embodiments, the nucleobase is selected from thymine, cytosine, guanine, adenine, uracil, and analogs or derivatives thereof.

[0115] In some embodiments, the sense strand or the antisense strand has the following chemical structure: [ka] wherein R x is a nucleobase. In some embodiments, the nucleobase is selected from thymine, cytosine, guanine, adenine, uracil, and analogs or derivatives thereof.

[0116] In some embodiments, the sense strand or the antisense strand has the following chemical structure: [ka] TIFF2024533262000008.tif54161, wherein B and R y is a nucleobase. In some embodiments, the nucleobase is selected from thymine, cytosine, guanine, adenine, uracil, and analogs or derivatives thereof.

[0117] In some embodiments, any sense or antisense nucleotide sequence described herein comprises, consists of, or consists essentially of ribonucleic acid (RNA). In some embodiments, any sense or antisense nucleotide sequence described herein comprises, consists of, or consists essentially of modified RNA. In some embodiments, the modified RNA is selected from 2'-O-methyl RNA and 2'-fluoro RNA. In some embodiments, 15, 16, 17, 18, 19, 20, 21, 22, or 23 modified nucleotides of any sense or antisense nucleotide sequence described herein are independently selected from 2'-O-methyl RNA and 2'-fluoro RNA.

[0118] In some embodiments, the siRNA molecules disclosed herein comprise terminal modifications at the 5'-end and / or 3'-end of the sense strand and / or antisense strand. In some embodiments, the siRNA molecules disclosed herein comprise a phosphate moiety at the 5'-end of the sense strand and / or antisense strand. In some embodiments, the 5'-end of the sense strand and / or antisense strand comprises a phosphate mimetic or analog (e.g., a "5'-end phosphate mimetic"). In some embodiments, the 5'-end of the sense strand and / or antisense strand comprises a vinyl phosphonate or a variant thereof (e.g., a "5'-end vinyl phosphonate").

[0119] In some embodiments, the siRNA molecule comprises at least one backbone modification, such as modified internucleoside linkage.In some embodiments, the siRNA molecule described herein comprises at least one phosphorothioate internucleoside linkage.In certain embodiments, the phosphorothioate internucleoside linkage can be located at the 3'-end or 5'-end of the sense strand and / or antisense strand.

[0120] In some embodiments, the siRNA molecule comprises at least one unpaired nucleotide overhang. In some embodiments, when the siRNA molecule comprises a nucleotide overhang, two or more unpaired nucleotides in the overhang can be linked by phosphorothioate internucleoside linkage. In certain embodiments, all unpaired nucleotides in the 3'-end nucleotide overhang of the antisense strand and / or sense strand are linked by phosphorothioate internucleoside linkage. In some embodiments, all unpaired nucleotides in the 5'-end nucleotide overhang of the antisense strand and / or sense strand are linked by phosphorothioate internucleoside linkage. In some embodiments, all unpaired nucleotides in any nucleotide overhang are linked by phosphorothioate internucleoside linkage.

[0121] In some embodiments, the sense strand or antisense strand may further comprise at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 or more phosphorothioate internucleoside linkages. In some embodiments, the sense strand comprises no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, or 3 phosphorothioate internucleoside linkages. In some embodiments, the sense strand comprises 2 to 10, 2 to 8, 2 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2 phosphorothioate internucleoside linkages. In some embodiments, the sense strand comprises 1 to 2 phosphorothioate internucleoside linkages. In some embodiments, the sense strand comprises 2 to 4 phosphorothioate internucleoside linkages. In some embodiments, at least one phosphorothioate internucleoside linkage is present between the 1st and 2nd nucleotides from the 5'-end of any sense or antisense nucleotide sequence described herein. In some embodiments, at least one phosphorothioate internucleoside linkage is present between the 2nd and 3rd nucleotides from the 5'-end of any sense or antisense nucleotide sequence described herein. In some embodiments, the sense strand comprises two phosphorothioate internucleoside linkages between the 1st to 3rd nucleotides from the 5'-end of any sense or antisense nucleotide sequence described herein.

[0122] In some embodiments, modified nucleotides that can be incorporated into the siRNA molecules of the present disclosure can have one or more chemical modifications described herein. For example, in some embodiments, the modified nucleotides can have modifications to the ribose sugar as well as modifications to the phosphodiester backbone. As an example, the modified nucleotides can include a 2' sugar modification (e.g., 2'-fluoro or 2'-O-methyl) and a modification to the 5' phosphate that results in a modified internucleoside linkage when the modified nucleotide is incorporated into a polynucleotide. For example, in some embodiments, the modified nucleotides can include a sugar modification (e.g., 2'-fluoro or 2'-O-methyl) and a 5' phosphorothioate group. In some embodiments, the sense and / or antisense strands of the siRNA molecules of the present disclosure include a combination of a 2' modified nucleotide and a phosphorothioate internucleoside linkage. In some embodiments, the sense and / or antisense strands of the siRNA molecules of the present disclosure include a combination of a 2' sugar modification, a phosphorothioate internucleoside linkage, and a 5' terminal vinyl phosphonate.

[0123] In some embodiments, any of the siRNAs disclosed herein contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more modified nucleotides. In some embodiments, any of the siRNAs disclosed herein contain one or more modified nucleotides. In some embodiments, any of the siRNAs disclosed herein contain two or more modified nucleotides. In some embodiments, any of the siRNAs disclosed herein contain five or more modified nucleotides. In some embodiments, any of the siRNAs disclosed herein contain eight or more modified nucleotides. In some embodiments, any of the siRNAs disclosed herein contain ten or more modified nucleotides. In some embodiments, any of the siRNAs disclosed herein contain 15 or more modified nucleotides. In some embodiments, any of the siRNAs disclosed herein contain 20 or more modified nucleotides. In some embodiments, any of the siRNAs disclosed herein contain 30 or more modified nucleotides. In some embodiments, any of the siRNAs disclosed herein contain 35 or more modified nucleotides. In some embodiments, any of the siRNAs disclosed herein contain 40 or more modified nucleotides. In some embodiments, any of the siRNAs disclosed herein contain 45 or more modified nucleotides. In some embodiments, all of the nucleotides in the siRNA molecule are modified nucleotides. In some embodiments, the one or more modified nucleotides are independently selected from 2'-O-methyl nucleotides, 2'-fluoro nucleotides, locked nucleic acids, nucleoside analogs, 5'-terminal vinyl phosphonates, and 5' phosphorothioates.

[0124] In some embodiments, any of the sense strands disclosed herein contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more modified nucleotides. In some embodiments, any of the sense strands disclosed herein contain one or more modified nucleotides. In some embodiments, any of the sense strands disclosed herein contain two or more modified nucleotides. In some embodiments, any of the sense strands disclosed herein contain five or more modified nucleotides. In some embodiments, any of the sense strands disclosed herein contain eight or more modified nucleotides. In some embodiments, any of the sense strands disclosed herein contain ten or more modified nucleotides. In some embodiments, any of the sense strands disclosed herein contain 15 or more modified nucleotides. In some embodiments, any of the sense strands disclosed herein contain 17 or more modified nucleotides. In some embodiments, any of the sense strands disclosed herein contain 18 or more modified nucleotides. In some embodiments, any of the sense strands disclosed herein contain 19 or more modified nucleotides. In some embodiments, any of the sense strands disclosed herein contain 20 or more modified nucleotides. In some embodiments, any of the sense strands disclosed herein contain 21 or more modified nucleotides. In some embodiments, all of the nucleotides in the sense strand are modified nucleotides. In some embodiments, the one or more modified nucleotides are independently selected from 2'-O-methyl nucleotides, 2'-fluoro nucleotides, locked nucleic acids, nucleoside analogs, 5'-terminal vinyl phosphonates, and 5' phosphorothioates.

[0125] In some embodiments, any of the antisense strands disclosed herein contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more modified nucleotides. In some embodiments, any of the antisense strands disclosed herein contain one or more modified nucleotides. In some embodiments, any of the antisense strands disclosed herein contain two or more modified nucleotides. In some embodiments, any of the antisense strands disclosed herein contain five or more modified nucleotides. In some embodiments, any of the antisense strands disclosed herein contain eight or more modified nucleotides. In some embodiments, any of the antisense strands disclosed herein contain ten or more modified nucleotides. In some embodiments, any of the antisense strands disclosed herein contain 15 or more modified nucleotides. In some embodiments, any of the antisense strands disclosed herein contain 17 or more modified nucleotides. In some embodiments, any of the antisense strands disclosed herein contain 18 or more modified nucleotides. In some embodiments, any of the antisense strands disclosed herein contain 19 or more modified nucleotides. In some embodiments, any of the antisense strands disclosed herein contain 20 or more modified nucleotides. In some embodiments, any of the antisense strands disclosed herein contain 21 or more modified nucleotides. In some embodiments, any of the antisense strands disclosed herein contain 22 or more modified nucleotides. In some embodiments, any of the antisense strands disclosed herein contain 23 or more modified nucleotides. In some embodiments, all of the nucleotides in the antisense strand are modified nucleotides. In some embodiments, the one or more modified nucleotides are independently selected from 2'-O-methyl nucleotides, 2'-fluoro nucleotides, locked nucleic acids, nucleoside analogs, 5'-terminal vinyl phosphonates, and 5' phosphorothioates.

[0126] In some embodiments, at least about 10%, 20%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 90%, 95%, or 100% of the nucleotides in any of the sense strands disclosed herein are modified nucleotides. In some embodiments, at least about 10% of the nucleotides in any of the sense strands disclosed herein are modified nucleotides. In some embodiments, at least about 30% of the nucleotides in any of the sense strands disclosed herein are modified nucleotides. In some embodiments, at least about 50% of the nucleotides in any of the sense strands disclosed herein are modified nucleotides. In some embodiments, at least about 60% of the nucleotides in any of the sense strands disclosed herein are modified nucleotides. In some embodiments, at least about 70% of the nucleotides in any of the sense strands disclosed herein are modified nucleotides. In some embodiments, at least about 80% of the nucleotides in any of the sense strands disclosed herein are modified nucleotides. In some embodiments, at least about 90% of the nucleotides in any of the sense strands disclosed herein are modified nucleotides. In some embodiments, at least about 100% of the nucleotides in any of the sense strands disclosed herein are modified nucleotides. In some embodiments, the one or more modified nucleotides are independently selected from 2'-O-methyl nucleotides, 2'-fluoro nucleotides, locked nucleic acids, nucleoside analogs, 5'-terminal vinyl phosphonates, and 5' phosphorothioates.

[0127] In some embodiments, at least about 10%, 20%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 90%, 95%, or 100% of the nucleotides in any of the antisense strands disclosed herein are modified nucleotides. In some embodiments, at least about 10% of the nucleotides in any of the antisense strands disclosed herein are modified nucleotides. In some embodiments, at least about 30% of the nucleotides in any of the antisense strands disclosed herein are modified nucleotides. In some embodiments, at least about 50% of the nucleotides in any of the antisense strands disclosed herein are modified nucleotides. In some embodiments, at least about 60% of the nucleotides in any of the antisense strands disclosed herein are modified nucleotides. In some embodiments, at least about 70% of the nucleotides in any of the antisense strands disclosed herein are modified nucleotides. In some embodiments, at least about 80% of the nucleotides in any of the antisense strands disclosed herein are modified nucleotides. In some embodiments, at least about 90% of the nucleotides in any of the antisense strands disclosed herein are modified nucleotides. In some embodiments, at least about 100% of the nucleotides in any of the antisense strands disclosed herein are modified nucleotides. In some embodiments, the one or more modified nucleotides are independently selected from 2'-O-methyl nucleotides, 2'-fluoro nucleotides, locked nucleic acids, nucleoside analogs, 5'-terminal vinyl phosphonates, and 5' phosphorothioates.

[0128] siRNA conjugates In some embodiments, the siRNA molecules disclosed herein may contain one or more conjugates or ligands. As used herein, "conjugate" or "ligand" refers to any compound or molecule that can directly or indirectly interact with another compound or molecule. In some embodiments, the ligand can modify one or more properties of the siRNA molecule to which it is bound, such as the pharmacodynamics, pharmacokinetics, binding, absorption, cellular distribution, cellular uptake, charge, and / or clearance properties of the siRNA molecule. Non-limiting examples of such conjugates are described, for example, in: WO2020 / 243490; WO2020 / 097342; WO2021 / 119325; PCT / US2021 / 019629; PCT / US2021 / 019628; PCT / US2021 / 021199; Sig. Transduct. Target Ther. 5 (101), 2020; ACS Chem. Biol. 10 (5), 1181-1187, 2015; J. Am. Chem. Soc. 136 (49), 16958-16961, 2014; Nucleic Acids Res. 42 (13), 8796-8807, 2014; Molec. Ther. 28 (8), 1759-1771, 2020; and Nucleic Acid Ther. 28 (3), 109-118, 2018; each of which is incorporated herein by reference.

[0129] In some embodiments, the ligand can be attached to the 5'-end and / or 3'-end of the sense strand and / or antisense strand of siRNA, for example, via a covalent bond to nucleotide.In some embodiments, the ligand is covalently attached to the sense strand or antisense strand of siRNA molecule via a linker.The ligand can be attached to the nucleobase, sugar moiety, or internucleoside linkage of the polynucleotide (e.g., sense strand or antisense strand) of the siRNA molecule of the present disclosure.

[0130] In some embodiments, the type of conjugate or ligand used and the degree of conjugation of the siRNA molecules of the present disclosure can be evaluated, for example, for improved pharmacokinetic profile, bioavailability, and / or stability of the siRNA molecule, while maintaining the ability of the siRNA to mediate RNAi activity. In some embodiments, the conjugate or ligand alters the distribution, targeting, or lifespan of the siRNA molecule into which it is incorporated. In some embodiments, the conjugate or ligand provides enhanced affinity for a selected target, for example, a molecule, cell or cell type, compartment (e.g., cellular compartment or organ compartment), tissue, organ, or body region, for example, compared to a molecule in the absence of such a ligand.

[0131] In some embodiments, the conjugate or ligand may comprise a naturally occurring substance or a recombinant or synthetic molecule.Non-limiting examples of conjugates and ligands include serum proteins (e.g., human serum albumin, low density lipoprotein, globulin), cholesterol moieties, vitamins (e.g., biotin, vitamin E, vitamin B12), folate moieties, steroids, bile acids (e.g., cholic acid), fatty acids (e.g., palmitic acid, myristic acid), carbohydrates (e.g., dextran, pullulan, chitin, chitosan, inulin, cyclodextrin, hyaluronic acid, or N-acetyl-galactosamine (Gal NAc), glycosides, phospholipids, antibodies or binding fragments thereof (e.g., antibodies or binding fragments that target siRNA to specific cell types such as the liver), dyes, intercalating agents (e.g., acridine), crosslinking agents (e.g., psoralens, mitomycin C), porphyrins (TPPC4, texaphyrin, sapphyrin), polycyclic aromatic hydrocarbons (e.g., phenazine, dihydrophenazine), artificial endonucleases (e.g., EDTA), lipophilic molecules (e.g., cholesterol, tocopherol, long-chain fatty acids (e.g., docosanoic acid), , palmitoyl, docosahexaenoic acid), cholic acid, adamantaneacetic acid, 1-pyrenebutyric acid, dihydrotestosterone, 1,3-bis-O(hexadecyl)glycerol, geranyloxyhexyl group, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecyl group, O3-(oleoyl)lithocholic acid, O3-(oleoyl)cholinic acid, dimethoxytrityl, or phenoxazine), peptides (e.g., antennapedia peptide, Tat peptide, RGD peptide), alkylating agents, polymers, e.g. For example, polyethylene glycol (PEG) (e.g., PEG-40K), polyamino acids, polyamines (e.g., spermine, spermidine), alkyls, substituted alkyls, radiolabeled markers, enzymes, haptens (e.g., biotin), transport / absorption enhancers (e.g., aspirin, vitamin E, folic acid), synthetic ribonucleases (e.g., imidazole, bisimidazole, histamine, imidazole clusters, acridine-imidazole conjugates, Eu3+ complexes of tetraazamacrocycles), dinitrophenyl, HRP, or AP.

[0132] In some embodiments, the conjugate or ligand comprises a carbohydrate, including, but not limited to, sugars (e.g., monosaccharides, disaccharides, trisaccharides, tetrasaccharides, and oligosaccharides containing about 4, 5, 6, 7, 8, or 9 monosaccharide units) and polysaccharides such as starch, glycogen, cellulose, and polysaccharide gums. In some embodiments, the carbohydrate incorporated into the ligand is a monosaccharide selected from pentose, hexose, or heptose, and disaccharides and trisaccharides containing such monosaccharide units.

[0133] In some embodiments, the carbohydrate incorporated into the conjugate or ligand is an amino sugar, such as galactosamine, glucosamine, N-acetyl-galactosamine (GalNAc), and N-acetyl-glucosamine. In some embodiments, the conjugate or ligand comprises N-acetyl-galactosamine and its derivatives. Non-limiting examples of GalNAc- or galactose-containing ligands that can be incorporated into the siRNAs of the present disclosure are described in the following: WO 2020 / 243490; WO 2020 / 097342; WO 2021 / 119325; PCT / US2021 / 019629; PCT / US2021 / 019628; PCT / US2021 / 021199; Sig. Transduct. Target Ther. 5 (101), 1-25, 2020; ACS Chem. Biol. 10 (5), 1181-1187, 2015; J. Am. Chem. Soc. 136 (49), 16958-16961, 2014; Nucleic Acids Res. 42 (13), 8796-8807, 2014; Molec. Ther. 28 (8), 1759-1771, 2020; and Nucleic Acid Ther. 28 (3), 109-118, 2018; all of which are incorporated herein by reference in their entireties.

[0134] The conjugate or ligand can be directly or indirectly bound or conjugated to the siRNA molecule. For example, in some embodiments, the ligand is directly covalently bound to the sense strand or antisense strand of the siRNA molecule. In other embodiments, the ligand is covalently bound to the sense strand or antisense strand of the siRNA molecule via a linker. The ligand can be bound to the nucleobase, sugar moiety, or internucleoside linkage of the polynucleotide (e.g., sense strand or antisense strand) of the siRNA molecule of the present disclosure. In some embodiments, the conjugate or ligand can be bound to the 5'-end and / or 3'-end of the sense strand and / or antisense strand of the siRNA molecule. In certain embodiments, the ligand is covalently bound to the 5'-end of the sense strand. In some embodiments, the ligand is covalently bound to the 3'-end of the sense strand. In some embodiments, the ligand is bound to the 5'-terminal nucleotide of the sense strand or the 3'-terminal nucleotide of the sense strand.

[0135] In some embodiments, the conjugate or ligand covalently attached to the sense strand and / or antisense strand of the siRNA molecule comprises a GalNAc derivative. In some embodiments, the GalNAc derivative is attached to the 5'-end and / or 3'-end of the sense strand and / or antisense strand of the siRNA molecule. In some embodiments, the GalNAc derivative is attached to the 3'-end of the sense strand. In some embodiments, the GalNAc derivative is attached to the 5'-end of the sense strand. In some embodiments, the GalNAc derivative is attached to the 3'-end of the antisense strand. In some embodiments, the GalNAc derivative is attached to the 5'-end of the antisense strand. In some embodiments, the GalNAc derivative is attached to the 5'-end of the sense strand and the 3'-end of the sense strand.

[0136] In some embodiments, the conjugate or ligand is a GalNAc derivative comprising 1, 2, 3, 4, 5, or 6 monomeric GalNAc units. In some embodiments, the conjugate or ligand is a GalNAc derivative comprising 1 monomeric GalNAc unit. In some embodiments, the conjugate or ligand is a GalNAc derivative comprising 2 monomeric GalNAc units. In some embodiments, the conjugate or ligand is a GalNAc derivative comprising 3 monomeric GalNAc units. In some embodiments, the conjugate or ligand is a GalNAc derivative comprising 4 monomeric GalNAc units. In some embodiments, the conjugate or ligand is a GalNAc derivative comprising 5 monomeric GalNAc units. In some embodiments, the conjugate or ligand is a GalNAc derivative comprising 6 monomeric GalNAc units. In some embodiments, various amounts of monomeric GalNAc units are attached to the 5'-end and 3'-end of the sense strand. In some embodiments, varying amounts of monomeric GalNAc units are attached to the 5'-end and 3'-end of the antisense strand. In some embodiments, 1, 2, 3, 4, 5, or 6 monomeric GalNAc units are attached to the 5'-end of the sense strand. In some embodiments, 1, 2, 3, 4, 5, or 6 monomeric GalNAc units are attached to the 3'-end of the sense strand. In some embodiments, 1, 2, 3, 4, 5, or 6 monomeric GalNAc units are attached to the 5'-end of the antisense strand. In some embodiments, 1, 2, 3, 4, 5, or 6 monomeric GalNAc units are attached to the 3'-end of the antisense strand. In some embodiments, the same number of monomeric GalNAc units are attached to both the 5'-end and 3'-end of the sense strand. In some embodiments, the same number of monomeric GalNAc units are attached to both the 5'-end and 3'-end of the antisense strand. In some embodiments, different numbers of monomeric GalNAc units are attached to the 5' and 3' ends of the sense strand, hi some embodiments, different numbers of monomeric GalNAc units are attached to the 5' and 3' ends of the antisense strand.

[0137] In some embodiments, the double-stranded siRNA molecule of any one of siRNA duplex ID Nos. ds-siNA D1-D178 or mds-siNA MD1-MD178 further comprises a GalNAc derivative attached to the 5'-end and / or 3'-end of the sense strand and / or antisense strand of the siRNA molecule. In some embodiments, the double-stranded siRNA molecule selected from any one of the siRNA duplexes in Table 8 or Table 9 or Table 10 or Table 11 or Table 12 further comprises a GalNAc derivative attached to the 5'-end and / or 3'-end of the sense strand and / or antisense strand of the siRNA molecule.

[0138] HSD17B13 In some embodiments, any of the siRNAs disclosed herein specifically downregulate the expression of the HSD17B13 gene or a variant thereof. In some embodiments, any of the siRNAs disclosed herein specifically downregulate the expression of the HSD17B13 gene or a variant thereof in cells by at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, wherein the percentage of downregulation of expression is compared to cells not contacted with the siRNA. In some embodiments, any of the siRNAs disclosed herein specifically downregulate the expression of the HSD17B13 gene or a variant thereof in cells by at least about 30%, wherein the percentage of downregulation of expression is compared to cells not contacted with the siRNA. In some embodiments, any of the siRNAs disclosed herein specifically downregulate the expression of the HSD17B13 gene or a variant thereof in a cell by at least about 50%, where the percent downregulation of expression is compared to cells not contacted with the siRNA. In some embodiments, any of the siRNAs disclosed herein specifically downregulate the expression of the HSD17B13 gene or a variant thereof in a cell by at least about 60%, where the percent downregulation of expression is compared to cells not contacted with the siRNA. In some embodiments, any of the siRNAs disclosed herein specifically downregulate the expression of the HSD17B13 gene or a variant thereof in a cell by at least about 70%, where the percent downregulation of expression is compared to cells not contacted with the siRNA. In some embodiments, any of the siRNAs disclosed herein specifically downregulate the expression of the HSD17B13 gene or a variant thereof in a cell by at least about 75%, where the percent downregulation of expression is compared to cells not contacted with the siRNA.In some embodiments, any of the siRNAs disclosed herein specifically down-regulate the expression of the HSD17B13 gene or a variant thereof in a cell by at least about 80%, where the percent down-regulation of expression is compared to cells not contacted with the siRNA. In some embodiments, any of the siRNAs disclosed herein specifically down-regulate the expression of the HSD17B13 gene or a variant thereof in a cell by at least about 85%, where the percent down-regulation of expression is compared to cells not contacted with the siRNA. In some embodiments, any of the siRNAs disclosed herein specifically down-regulate the expression of the HSD17B13 gene or a variant thereof in a cell by at least about 90%, where the percent down-regulation of expression is compared to cells not contacted with the siRNA. In some embodiments, any of the siRNAs disclosed herein specifically down-regulate the expression of the HSD17B13 gene or a variant thereof in a cell by at least about 95%, where the percent down-regulation of expression is compared to cells not contacted with the siRNA. In some embodiments, any of the siRNAs disclosed herein specifically downregulates expression of the HSD17B13 gene or a variant thereof in a cell by at least about 100%, wherein the percent downregulation of expression is compared to a cell not contacted with the siRNA.

[0139] The expression of the HSD17B13 gene can be measured by any method known in the art. Exemplary methods for measuring the expression of the HSD17B13 gene include, but are not limited to, quantitative PCR, RT-PCR, RT-qPCR, Western blot, Southern blot, Northern blot, FISH, DNA microarray, tiling array, and RNA-Seq. The expression of the HSD17B13 gene can be evaluated, for example, based on the level or change in the level of any variable associated with the expression of the HSD17B13 gene (e.g., HSD17B13 mRNA level, HSD17B13 protein level, and / or the number or extent of amyloid deposits). This level can be evaluated in individual cells or groups of cells, including, for example, samples derived from a subject. In some embodiments, downregulation or inhibition can be evaluated by a decrease in the absolute or relative level of one or more variables associated with the expression of HSD17B13 compared to a control level. The control level can be any type of control level utilized in the art, e.g., a baseline level before administration, or a level determined from a similar subject, cell, or sample that is untreated or treated with a control (e.g., a buffer-only control, an inactive or attenuated agent control, etc.).

[0140] In some embodiments, the HSD17B13 gene comprises a nucleotide sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleotide sequence of SEQ ID NO:1 (GenBank Accession No. NM_178135.5 (nucleotides 42-944)) over the entire length of SEQ ID NO:261.

[0141] In some embodiments, the HSD17B13 gene comprises a nucleotide sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 nucleotide mismatches to the nucleotide sequence of SEQ ID NO:261 over the entire length of SEQ ID NO:261.

[0142] In some embodiments, the fragment of the HSD17B13 gene is about 10 to about 50, or about 15 to about 50, or about 15 to about 45 nucleotides in length, or about 15 to about 40, or about 15 to about 35, or about 15 to about 30, or about 15 to about 25, or about 17 to about 23 nucleotides, or about 17 to about 22, or about 17 to about 21, or about 18 to about 23, or about 18 to about 22, or about 18 to about 21, or about 19 to about 23, or about 19 to about 22, or about 19 to about 21 nucleotides in length.

[0143] Administration of siRNA Administration of any of the siRNAs disclosed herein can be carried out by methods known in the art, including the following: The siRNAs of the present disclosure can be administered systemically or locally, for example, orally, nasally, parenterally, topically, intracisternally, intravaginally, or intrarectally, and are administered in a form suitable for each administration route.

[0144] Delivery of the siRNA molecules of the present disclosure to cells, e.g., cells of a subject (e.g., a human subject, e.g., a subject in need thereof, e.g., a subject with a disease, disorder, or condition associated with HSD17B13 gene expression) can be achieved in many different ways. For example, in some embodiments, delivery can be performed by contacting cells with the siRNA of the present disclosure either in vitro, in vivo, or ex vivo. In some embodiments, in vivo delivery can be performed, for example, by administering a pharmaceutical composition comprising the siRNA molecule to a subject. In some embodiments, in vivo delivery can be performed by administering one or more vectors encoding the siRNA and inducing its expression.

[0145] Generally, any method of delivering nucleic acid molecules (in vitro, in vivo or ex vivo) can be adapted for use with the siRNA molecules of the present disclosure.For in vivo delivery, factors to be considered for delivering siRNA molecules include, for example, the biological stability of the delivered molecule, prevention of non-specific effects, and the accumulation of the delivered molecule in target tissue and non-target tissue.

[0146] In some embodiments, the non-specific effects of siRNA can be minimized by local administration, for example, by direct injection or implantation into tissue, or by local administration of a preparation. Local administration at the treatment site can, for example, maximize the local concentration of the agent, limit exposure of the agent to systemic tissues that may otherwise be harmed by or degrade the agent, and allow for the administration of a lower total dose of siRNA molecules.

[0147] In some embodiments, the siRNAs or pharmaceutical compositions comprising the siRNAs of the present disclosure, with or without being incorporated into biopolymers, can be administered locally to the relevant tissue, ex vivo or in vivo, for example, via injection, infusion pump, or stent.

[0148] When siRNA is administered to treat disease, siRNA can be modified or alternatively can be delivered using drug delivery system; either method can act, for example, to prevent the rapid degradation of dsRNA by endonuclease and exonuclease in vivo.The modification of pharmaceutical carrier or siRNA can also enable the targeting of siRNA composition to target tissue, and can avoid undesirable off-target effects.For example, siRNA molecule can be modified by conjugation with lipophilic group such as cholesterol as mentioned above, thereby, for example, can promote uptake into cell and prevent degradation.

[0149] In some embodiments, the siRNA can be delivered using a drug delivery system such as a nanoparticle, a dendrimer, a polymer, a liposome, or a cationic delivery system. A positively charged cationic delivery system can promote the binding of siRNA molecules (which are negatively charged) and can also enhance the interaction with the negatively charged cell membrane, allowing for efficient uptake of siRNA by cells. In some embodiments, a cationic lipid, a dendrimer, or a polymer can be bound to the siRNA or induced to form a vesicle or micelle that encapsulates the siRNA. The formation of a vesicle or micelle can also prevent the degradation of the siRNA, for example, when administered systemically.

[0150] Non-limiting examples of drug delivery systems useful for systemic delivery of siRNA include DOTAP, cardiolipin, polyethyleneimine, Arg-Gly-Asp (RGD) peptide, and polyamidoamine. In some embodiments, the siRNA is complexed with cyclodextrin for systemic administration.

[0151] Pharmaceutical Composition The siRNA molecules of the present disclosure can be administered as pharmaceuticals to animals, including mammals, particularly humans, by themselves, in mixtures with one another, and / or in the form of pharmaceutical compositions.

[0152] The present disclosure encompasses pharmaceutical compositions and preparations comprising the siRNA molecules of the present disclosure.In some embodiments, the siRNA molecules of the present disclosure can be administered in pharmaceutical compositions.In some embodiments, the pharmaceutical compositions of the present disclosure comprise one or more siRNA molecules of the present disclosure and pharmaceutically acceptable carriers.It should be understood that when referring to siRNA molecules in the present disclosure, if appropriate, the pharmaceutical compositions comprising siRNA molecules are also referred to.

[0153] In some embodiments, the pharmaceutical composition comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more of any of the siRNA molecules disclosed herein.

[0154] In some embodiments, any of the pharmaceutical compositions disclosed herein comprises one or more excipients, carriers, wetting agents, diluents, emulsifiers, lubricants, colorants, release agents, coating agents, sweetening agents, flavoring and perfuming agents, preservatives, and antioxidants.

[0155] In some embodiments, the siRNA molecules of the present disclosure can be administered in a "naked" form, where modified or unmodified siRNA molecules are directly suspended in aqueous or suitable buffer as "free siRNA". The free siRNA can be present in a suitable buffer containing, for example, acetate, citrate, prolamin, carbonate, or phosphate, or any combination thereof. In one embodiment, the buffer is phosphate-buffered saline (PBS). The pH and osmolality of the buffer containing the siRNA can be adjusted to be suitable for administration to a subject.

[0156] Examples of pharmaceutically acceptable antioxidants include, but are not limited to, the following: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, and the like; and (3) metal chelating agents, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0157] In certain embodiments, a pharmaceutical composition of the present disclosure comprises an excipient selected from the group consisting of cyclodextrins, celluloses, liposomes, micelle-forming agents (e.g., bile acids), and polymeric carriers (e.g., polyesters and polyanhydrides); and a compound of the present disclosure (e.g., an siRNA molecule). In certain embodiments, the composition renders the siRNA molecule of the present disclosure orally bioavailable.

[0158] Methods of preparing these formulations or pharmaceutical compositions include, for example, the step of bringing into association the siRNA molecules of this disclosure with the carrier and, optionally, one or more accessory ingredients. Generally, the formulations are prepared by uniformly and intimately bringing into association the siRNA molecules of this disclosure with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0159] The pharmaceutical compositions of the present disclosure can be administered via any common route, and are administered in a form suitable for each administration route. Such routes include, but are not limited to, parenteral (e.g., subcutaneous, intramuscular, intraperitoneal, or intravenous), oral, nasal, respiratory (e.g., aerosol), buccal, intradermal, transdermal, sublingual, rectal, and vaginal. In some embodiments, administration is by direct injection into liver tissue or delivery via the hepatic portal vein. In some embodiments, the pharmaceutical composition is administered orally. In some embodiments, the pharmaceutical composition is administered parenterally. In some embodiments, the composition is administered by subcutaneous or intravenous infusion or injection. In some embodiments, the pharmaceutical composition is administered subcutaneously.

[0160] Pharmaceutical compositions of the present disclosure suitable for oral administration can be present, for example, in the form of capsules (e.g., hard or soft capsules), cachets, pills, tablets, lozenges (using a flavored base, usually, for example, sucrose and acacia or tragacanth), powder, granules, or as a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as a pastille (using an inert base, such as gelatin and glycerin, or sucrose and acacia), and / or as a mouthwash, each containing a predetermined amount of the siRNA molecule of the present disclosure as an active ingredient. The siRNA molecule of the present disclosure can also be administered as a bolus, electuary, or paste.

[0161] In solid dosage forms of the present disclosure for oral administration (such as capsules, tablets, pills, dragees, powders, granules, lozenges, etc.), the active ingredient is mixed with one or more pharmaceutically acceptable carriers (e.g., sodium citrate or dicalcium phosphate) and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; and (5) solution retarding agents. (6) absorption enhancers, such as quaternary ammonium compounds, and surfactants, such as poloxamers and sodium lauryl sulfate; (7) wetting agents, such as cetyl alcohol, glycerol monostearate, and nonionic surfactants; (8) adsorbents, such as kaolin and bentonite clay; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, zinc stearate, sodium stearate, stearic acid, and mixtures thereof; (10) coloring agents; and (11) controlled-release agents, such as crospovidone or ethylcellulose.

[0162] In the case of capsules, tablets, and pills, the pharmaceutical compositions may also contain buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-shell gelatin capsules using such excipients as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like.

[0163] Tablets can be made, for example, by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared using, for example, binders (e.g., gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), surface-active agents, or dispersing agents. Molded tablets can be made, for example, by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

[0164] Tablets and other solid dosage forms (e.g., dragees, capsules, pills, and granules) of the pharmaceutical compositions of the present disclosure can optionally be scored or prepared with coatings and shells (e.g., enteric coatings and other coatings well known in the pharmaceutical formulating art). They can also be formulated to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropyl methylcellulose, other polymer matrices, liposomes, and / or microspheres, proportioned to provide the desired release profile. They can also be formulated for rapid release (e.g., lyophilized, etc.).

[0165] They can be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved in sterile water or other sterile injectable medium immediately before use. These compositions can also optionally contain opacifying agents and can be of a composition that releases the active ingredient only, or preferentially, in a certain part of the gastrointestinal tract, optionally in a delayed manner. Examples of implantable compositions that can be used include polymeric substances and waxes. The active ingredient can also be in microencapsulated form, if appropriate, with one or more of the above-mentioned excipients.

[0166] Liquid dosage forms for oral administration of the siRNA molecules of the present disclosure include, for example, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.In addition to the active ingredient, the liquid dosage forms can contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (such as cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan and their mixtures.

[0167] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0168] Suspensions can contain, in addition to the siRNA molecules, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, and tragacanth, and mixtures thereof.

[0169] Formulations of pharmaceutical compositions of the present disclosure for rectal or vaginal administration can be provided as suppositories, which can be prepared by mixing one or more siRNA molecules of the present disclosure with one or more suitable non-irritating excipients or carriers, including, for example, cocoa butter, polyethylene glycol, a suppository wax, or a salicylate, and which are, for example, solid at room temperature but liquid at body temperature, and therefore will melt in the rectum or vaginal cavity to release the siRNA molecules.

[0170] Formulations of the present disclosure which are suitable for vaginal administration include, for example, pessaries, tampons, creams, gels, pastes, foams, or spray formulations containing such carriers as are known in the art to be appropriate.

[0171] Dosage forms for topical or transdermal administration of the siRNA molecules of the present disclosure include, for example, powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The siRNA molecules can be mixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives, buffers, or propellants.

[0172] The ointments, pastes, creams, and gels can contain, in addition to the active siRNA molecules of the present disclosure, excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc, and zinc oxide, or mixtures thereof.

[0173] Powders and sprays can contain, in addition to the siRNA molecules of the present disclosure, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons (e.g., butane and propane).

[0174] Transdermal patch has the additional advantage of providing the siRNA molecule of the present disclosure to the body in a controlled manner.Such dosage form can be prepared by dissolving or dispersing the siRNA molecule in suitable medium.Absorption enhancers can also be used to increase the flux of siRNA molecule across skin.The rate of such flux can be controlled, for example, by providing a rate-controlling membrane or dispersing the siRNA molecule in a polymer matrix or gel.

[0175] Pharmaceutical compositions of the present disclosure suitable for parenteral administration comprise one or more siRNA molecules of the present disclosure in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions or sterile powders (which can be reconstituted into sterile injectable solutions or dispersions immediately before use), which may contain, for example, sugars, alcohols, antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0176] Examples of suitable aqueous and non-aqueous carriers that can be employed in the pharmaceutical compositions of the present disclosure include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0177] The pharmaceutical compositions of the present disclosure may further contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of microbial action on the target compounds can be ensured, for example, by including various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars and sodium chloride, in the compositions. Furthermore, prolonged absorption of the injectable pharmaceutical form can be achieved by including agents that delay absorption (e.g., aluminum monostearate and gelatin).

[0178] In some embodiments, to prolong the effect of a drug, it is desirable to slow the absorption of the drug, for example, from subcutaneous or intramuscular injection. This can be accomplished, for example, by using a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the drug depends on its rate of dissolution, which may depend on crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form can be accomplished by dissolving or suspending the drug in an oil vehicle.

[0179] In some embodiments, the administration is carried out via depot injection.Injectable depot forms can be prepared by forming a microencapsule matrix of the target siRNA molecule in a biodegradable polymer such as polylactide-polyglycolide.Depending on the ratio of drug to polymer and the properties of the specific polymer used, the release rate of the drug can be controlled.Other examples of biodegradable polymers include poly(orthoester) and poly(anhydride).Injectable depot preparations can also be prepared by encapsulating the drug in liposomes or microemulsions that are compatible with body tissues, for example.

[0180] Depot injections can release siRNA in a consistent manner over an extended period of time. Thus, depot injections can reduce the frequency of administration required to achieve a desired effect (e.g., a desired inhibition of HSD17B13, or a therapeutic or prophylactic effect). Depot injections can also provide a more stable serum concentration. Depot injections can include, for example, subcutaneous injections or intramuscular injections. In some embodiments, the depot injection is a subcutaneous injection.

[0181] In some embodiments, administration is carried out via a pump. The pump can be an external pump or a surgically implanted pump. In certain embodiments, the pump is a subcutaneously implanted osmotic pump. In another embodiment, the pump is an infusion pump. The infusion pump can be used for, for example, intravenous infusion, subcutaneous infusion, arterial infusion, or epidural infusion. In some embodiments, the infusion pump is a subcutaneous infusion pump. In another embodiment, the pump is a surgically implanted pump that delivers siRNA to the subject.

[0182] In some embodiments, the pharmaceutical compositions of the present disclosure are packaged with or stored within a device for administration. Devices for injectable formulations include, but are not limited to, injection ports, prefilled syringes, auto-injectors, injection pumps, on-body syringes, and injection pens. Devices for aerosolized or powder formulations include, but are not limited to, inhalers, insufflators, and aspirators. Thus, the present disclosure encompasses administration devices comprising the pharmaceutical compositions of the present disclosure for treating or preventing one or more of the disorders described herein.

[0183] The method of administration can be selected based on, for example, whether local or systemic treatment is desired and based on the area to be treated. The route and site of administration can be selected, for example, to facilitate targeting.

[0184] Regardless of the selected route of administration, the siRNA molecules of the present disclosure (which may be used in a suitable hydrated form) and / or pharmaceutical compositions of the present disclosure can be formulated into a pharmaceutically acceptable dosage form by methods known to those skilled in the art. The method for formulating a pharmaceutical composition depends on many criteria, including but not limited to the route of administration, the type and severity of the disease or disorder to be treated, and / or the dose to be administered. In some embodiments, the pharmaceutical composition is formulated based on the intended delivery route. Preparation of pharmaceutical compositions can be carried out by known methods. For this purpose, one or more compounds are combined with one or more solid or liquid pharmaceutical carrier substances and / or additives (or auxiliary substances), if necessary, with other pharmaceutically active compounds having a therapeutic or prophylactic effect, to form an appropriate dosage form or dosage.

[0185] The pharmaceutical compositions can be conveniently provided in unit dosage form and can be prepared by any method known in the art of pharmacy. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form varies depending on the host being treated and the particular method of administration, for example, as described below. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form is generally, for example, the amount of siRNA molecules that produces a therapeutic effect. In some embodiments, for example, out of 100 percent, this amount will range from about 0.1 percent to about 99 percent of the active ingredient, or from about 5 percent to about 70 percent, or from about 10 percent to about 30 percent.

[0186] The actual dosage level of the active ingredient in the pharmaceutical composition of the present disclosure can be varied to obtain an amount of the active ingredient that is effective for achieving the desired therapeutic response for a particular patient, composition, and administration method without causing toxicity to the patient.For example, the siRNA molecule in the pharmaceutical composition of the present disclosure can be administered at a dosage sufficient to downregulate the expression of the HSD17B13 gene.

[0187] The siRNA molecules and pharmaceutical compositions of the present disclosure can be used to treat a disease in a subject in need thereof, for example, in the methods described below.

[0188] Dosage The regimen and / or dosage utilizing the siRNA molecules of the present disclosure can be selected according to a variety of factors, including, for example, the activity of the particular siRNA molecule or salt thereof employed, the severity of the condition being treated, the route of administration, the time of administration, the rate of excretion or metabolism of the particular siRNA molecule employed, the rate and extent of absorption, the duration of treatment, other drugs, compounds and / or substances used in combination with the particular siRNA molecule employed, the type, breed, age, sex, weight, condition, general health and medical history of the patient being treated, the patient's renal and hepatic function, and factors well known in the medical arts. Consideration of these factors is within the purview of an ordinarily skilled clinician for purposes of determining a therapeutically effective amount.

[0189] In some embodiments, the appropriate daily dose of the siRNA molecule of the present disclosure is, for example, the amount of the siRNA molecule that is the minimum effective dose to produce a therapeutic effect.For example, a doctor or veterinarian can start the dosage of the siRNA molecule of the present disclosure that is adopted in a pharmaceutical composition at a level lower than the level required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved.This effective dose can depend, for example, on the factors mentioned above.In some embodiments, the siRNA molecule of the present disclosure can be administered at a dose sufficient to downregulate or inhibit the expression of HSD17B13 gene.

[0190] In some embodiments, the siRNA molecule is administered at about 0.01 mg / kg to about 200 mg / kg, or about 0.1 mg / kg to about 100 mg / kg, or about 0.5 mg / kg to about 50 mg / kg, or about 1 mg / kg to about 40 mg / kg, or about 1 mg / kg to about 30 mg / kg, or about 1 mg / kg to about 20 mg / kg, or about 1 mg / kg to about 15 mg / kg, or about 1 mg / kg to about 10 mg / kg. In some embodiments, the siRNA molecule is administered at a dose equal to or greater than 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95 or 1 mg / kg. In some embodiments, the siRNA molecule is administered at a dose equal to or greater than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 mg / kg. In some embodiments, the siRNA molecule is administered at a dose equal to or less than 200, 190, 180, 170, 160, 150, 140, 130, 120, 110, 100, 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, or 15 mg / kg. In some embodiments, the total daily dose of the siRNA molecules is equal to or greater than 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195 or 100 mg.

[0191] In some embodiments, treatment of a subject with a therapeutically effective amount of the siRNA molecule of the present disclosure can include a single treatment or a series of treatments.In some embodiments, the siRNA molecule can be administered as a single dose or divided into multiple doses.In some embodiments, the effective daily dose of the siRNA molecule can be administered as 2, 3, 4, 5, 6, 7, 8, 9, 10 or more doses or subdoses that are administered separately at appropriate intervals throughout the day, optionally in a unit dosage form.

[0192] In some embodiments, the siRNA molecule is administered once a day. In some embodiments, the siRNA molecule is administered once a week. In some embodiments, the siRNA molecule is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 times a day. In some embodiments, the siRNA molecule is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 times a week. In some embodiments, the siRNA molecule is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 times per month. In some embodiments, the siRNA molecule is administered once every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 days. In some embodiments, the siRNA molecule is administered every three days. In some embodiments, the siRNA molecule is administered once every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 weeks. In some embodiments, the siRNA molecule is administered once every month. In some embodiments, the siRNA molecule is administered once every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 months.

[0193] In some embodiments, the siRNA molecule comprises at least 1, 2, 3, 4, 5, 6, 7, 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, and administering at least 1, 2, 3, 4, 5, 6, 7, 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, or 53 doses over a period of 62, 63, 64, 65, 66, 67, 68, 69, or 70 days. In some embodiments, the siRNA molecule comprises at least 1, 2, 3, 4, 5, 6, 7, 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, or 5 At least 1, 2, 3, 4, 5, 6, 7, 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 or 53 doses are administered over a 3 week period.In some embodiments, the siRNA molecule comprises at least 1, 2, 3, 4, 5, 6, 7, 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, or 5 At least 1, 2, 3, 4, 5, 6, 7, 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 or 53 doses are administered over a 3 month period. In some embodiments, the siRNA molecule is administered at least once a week for a period of at least 1, 2, 3, 4, 5, 6, 7, 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, or 70 weeks. In some embodiments, the siRNA molecule is administered at least once a week for a period of at least 1, 2, 3, 4, 5, 6, 7, 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, or 70 months.In some embodiments, the siRNA molecule is administered at least twice a week for a period of at least 1, 2, 3, 4, 5, 6, 7, 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, or 70 weeks. In some embodiments, the siRNA molecule is administered at least twice a week for a period of at least 1, 2, 3, 4, 5, 6, 7, 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, or 70 months. In some embodiments, the siRNA molecule is administered at least once every two weeks for a period of at least 2, 3, 4, 5, 6, 7, 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, or 70 weeks. In some embodiments, the siRNA molecule is administered at least once every two weeks for a period of at least 2, 3, 4, 5, 6, 7, 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, or 70 months.In some embodiments, the siRNA molecule is administered at least once every four weeks for a period of at least 4, 5, 6, 7, 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, or 70 weeks. In some embodiments, the siRNA molecule is administered at least once every four weeks for a period of at least 4, 5, 6, 7, 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, or 70 months.

[0194] In some embodiments, a repeat dosing regimen may involve administering a therapeutically effective amount of siRNA periodically (e.g., every other day, once a week, once a quarter (i.e., approximately once every three months), or once a year). In some embodiments, the dosage and / or frequency of administration may be reduced after an initial treatment period. In some embodiments, when an siRNA molecule described herein is co-administered with another active agent, the therapeutically effective amount may be less than when the siRNA molecule is used alone.

[0195] Methods and Uses Also disclosed herein is a method for treating an HSD17B13-related disease in a subject in need thereof, comprising administering to the subject any of the siRNA molecules and / or pharmaceutical compositions comprising the siRNA molecules disclosed herein. In embodiments, the HSD17B13-related disease is a liver disease.

[0196] When the siRNA molecule of the present disclosure is administered to humans and animals as a pharmaceutical, the siRNA molecule can be administered by itself, or can be administered as the above-mentioned pharmaceutical composition containing, for example, 0.1 to 99% (more preferably, 10 to 30%) of the siRNA molecule in combination with a pharmaceutically acceptable carrier.

[0197] In some embodiments, a method for treating a disease in a subject in need of such treatment comprises administering to the subject a certain amount of any of the siRNA molecules disclosed herein.In embodiments, the certain amount is a therapeutically effective amount.In some embodiments, a method for treating a disease in a subject in need of such treatment comprises administering to the subject a certain amount of any of the pharmaceutical compositions disclosed herein.In embodiments, the certain amount is a therapeutically effective amount.

[0198] In some embodiments, a method for treating a disease in a subject in need thereof comprises administering to the subject any of the siRNA molecules or pharmaceutical compositions disclosed herein in combination with an additional active agent. In some embodiments, the additional active agent is a liver disease treatment drug. In embodiments, the amount of the siRNA molecule is a therapeutically effective amount. In embodiments, the amount of the additional active agent is a therapeutically effective amount.

[0199] In some embodiments, the siRNA molecule and the liver disease therapeutic agent are administered separately. In some embodiments, the siRNA molecule or pharmaceutical composition and the liver disease therapeutic agent are administered simultaneously. In some embodiments, the siRNA molecule or pharmaceutical composition and the liver disease therapeutic agent are administered sequentially. In some embodiments, the siRNA molecule or pharmaceutical composition is administered before the liver disease therapeutic agent is administered. In some embodiments, the siRNA molecule or pharmaceutical composition is administered after the liver disease therapeutic agent is administered. In some embodiments, the pharmaceutical composition comprises the siRNA and the liver disease therapeutic agent.

[0200] Also disclosed herein is a method for reducing the expression level of HSD17B13 in a subject in need thereof, comprising administering to the subject an amount of an siRNA molecule or pharmaceutical composition according to the present disclosure. In embodiments, the amount of the additional active agent is a therapeutically effective amount. In some embodiments, the method for reducing the expression level of HSD17B13 in a subject in need thereof, comprising administering to the subject an amount of an siRNA molecule or pharmaceutical composition according to the present disclosure, reduces the expression level of HSD17B13 in liver cells of the subject after administration of the siRNA molecule or pharmaceutical composition, compared to the expression level of HSD17B13 in a patient who has not been administered the siRNA or pharmaceutical composition.

[0201] Also disclosed herein is a method for preventing at least one symptom of liver disease in a subject in need thereof, comprising administering to the subject a specific amount of any of the siRNA molecules or pharmaceutical compositions of the present disclosure, thereby preventing at least one symptom of liver disease in the subject. In embodiments, the amount of the additional active agent is a therapeutically effective amount.

[0202] In another aspect, the present disclosure also discloses the use of any of the siRNA molecules or pharmaceutical compositions of the present disclosure in the manufacture of a medicament for treating liver disease. In some embodiments, the present disclosure provides the use of an siRNA molecule of the present disclosure or a pharmaceutical composition comprising an siRNA of the present disclosure that targets the HSD17B13 gene in mammalian cells in the manufacture of a medicament for inhibiting the expression of the HSD17B13 gene in a mammal.

[0203] The methods and uses disclosed herein include administering to a mammal (e.g., a human) a pharmaceutical composition comprising an siRNA molecule that targets the HSD17B13 gene in the cells of the mammal, and maintaining the composition for a period of time sufficient to result in degradation of mRNA transcripts of the HSD17B13 gene, thereby inhibiting expression of the HSD17B13 gene in the mammal.

[0204] The patient or subject of the described methods can be a mammal, which includes human and non-human mammals, hi some embodiments, the subject is a human, such as an adult human, a human teenager, a human child, a human toddler, or a human infant.

[0205] The siRNA molecules and / or pharmaceutical compositions of the present disclosure can be administered in the disclosed methods and uses by any route of administration known in the art, including, for example, subcutaneous, intravenous, oral, intraperitoneal, or parenteral routes (which include, for example, intracranial (e.g., intraventricular, intraparenchymal, and intrathecal), intramuscular, transdermal, intratracheal (aerosol), intranasal, rectal, and topical (which includes buccal and sublingual) administration).

[0206] The siRNA molecules and / or pharmaceutical compositions of the present disclosure can be administered in the disclosed methods and uses in any of the dosages or administration regimens described above.

[0207] HSD17B13-related disorders Any of the siRNAs and / or pharmaceutical compositions and / or methods and / or uses disclosed herein can be used to treat diseases, disorders, and / or conditions. In some embodiments, the diseases, disorders, and / or conditions are associated with HSD17B13 expression or activity. In some embodiments, the diseases, disorders, and / or conditions are liver diseases. As used herein, the term "HSD17B13-associated diseases" encompasses diseases, disorders, or conditions that benefit from downregulation of HSD17B13 gene expression, replication, or activity. Non-limiting examples of HSD17B13-associated diseases include, but are not limited to, fatty liver (steatosis), non-alcoholic steatohepatitis (NASH), cirrhosis, liver fat accumulation, liver inflammation, hepatocellular necrosis, liver fibrosis, obesity, hepatocellular carcinoma (HCC), or non-alcoholic fatty liver disease (NAFLD). In an embodiment, the HSD17B13-associated disease is NAFLD. In an embodiment, the HSD17B13-associated disease is NASH. In an embodiment, the HSD17B13-associated disease is fatty liver (steatosis). In an embodiment, the HSD17B13-associated disease is NAFLD. In an embodiment, the HSD17B13-associated disease is HCC.

[0208] Combination therapy Any of the siRNAs or pharmaceutical compositions disclosed herein can be combined with one or more additional active agents in pharmaceutical compositions, or in any method according to the present disclosure, or for use in treating liver disease.An additional active agent is a component that exhibits pharmacological effects at appropriate doses.An additional active agent can be another siRNA according to the present disclosure, an siRNA not according to the present disclosure, or a non-siRNA active agent.

[0209] In some embodiments, at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more siRNAs disclosed herein are combined in the combination therapy.

[0210] In some embodiments, any of the siRNAs or pharmaceutical compositions disclosed herein is combined in combination therapy with a liver disease therapeutic agent, in some embodiments, the liver disease therapeutic agent is selected from a peroxisome proliferator-activated receptor (PPAR) agonist, a farnesoid X receptor (FXR) agonist, a lipid-altering agent, an incretin-based therapy, a PNPLA3 inhibitor, and a thyroid hormone receptor (THR) modulator.

[0211] In some embodiments, any of the siRNAs or pharmaceutical compositions disclosed herein is combined with a PPAR agonist. In some embodiments, the PPAR agonist is selected from a PPARα agonist, a dual PPARα / δ agonist, a PPARγ agonist, and a dual PPARα / γ agonist. In some embodiments, the dual PPARα agonist is a fibrate. In some embodiments, the PPARα / δ agonist is elafibranor. In some embodiments, the PPARγ agonist is a thiazolidinedione (TZD). In some embodiments, the TZD is pioglitazone. In some embodiments, the dual PPARα / γ agonist is saroglitazar.

[0212] In some embodiments, any of the siRNAs or pharmaceutical compositions disclosed herein is combined with an FXR agonist, in some embodiments, the FXR agonist is selected from obeticholic acid (OCA) and TERN-1010.

[0213] In some embodiments, any of the siRNAs or pharmaceutical compositions disclosed herein is combined with a lipid-altering agent, hi some embodiments, the lipid-altering agent is aramchol.

[0214] In some embodiments, any of the siRNAs or pharmaceutical compositions disclosed herein is combined with an incretin-based therapy. In some embodiments, the incretin-based therapy is a glucagon-like peptide 1 (GLP-1) receptor agonist or a dipeptidyl peptidase 4 (DPP-4) inhibitor. In some embodiments, the GLP-1 receptor agonist is exenatide or liraglutide. In some embodiments, the DPP-4 inhibitor is sitagliptin or vildapliptin.

[0215] In some embodiments, any of the siRNAs or pharmaceutical compositions disclosed herein is combined with a THR modulator. In some embodiments, the THR modulator is selected from a THR-β modulator and a thyroid hormone analog. Exemplary THR modulators are described in: Jakobsson, et al., Drugs, 2017, 77(15):1613-1621; Saponaro, et al., Front Med (Lausanne), 2020, 7:331; and Kowalik, et al., Front Endocrinol, 2018, 9:382; which are incorporated by reference in their entirety. In some embodiments, the THR-β modulator is a THR-β agonist. In some embodiments, the THR-β agonist is selected from KB141, sobetirome, Sob-AM2, eprotirome, VK2809, resmetirome, MB07344, IS25, TG68, GC-24, and any one of the compounds disclosed in U.S. Patent No. 11,091,467, which is incorporated herein by reference in its entirety. In some embodiments, the thyroid hormone analog is selected from L-94901 and CG-23425.

[0216] In general, the liver disease therapeutic agent can be used in any combination with the siRNA molecule of the present disclosure in a single dosage form (e.g., a fixed-dose pharmaceutical combination) or in one or more separate dosage forms that allow for simultaneous or sequential administration of the active agents (co-administration of separate active agents) to a subject. In some embodiments, the siRNA and the liver disease therapeutic agent are administered simultaneously. In some embodiments, the siRNA and the liver disease therapeutic agent are administered sequentially. In some embodiments, the siRNA is administered before the liver disease therapeutic agent. In some embodiments, the siRNA is administered after the liver disease therapeutic agent. The order and frequency in which the siRNA and the liver disease therapeutic agent are administered can vary. In some embodiments, the siRNA and the liver disease therapeutic agent are present in separate containers. In some embodiments, the siRNA and the liver disease therapeutic agent are present in the same container. In some embodiments, the pharmaceutical composition comprises the siRNA and the liver disease therapeutic agent. The siRNA and the liver disease therapeutic agent can be administered via the same or different routes of administration.

[0217] Yet another embodiment of the present disclosure has the following features.

[0218] The present technology provides short interfering nucleic acid (siNA) molecules. The siNA can be single-stranded. Alternatively, the siNA can be a double-stranded (ds-siNA) molecule. In any embodiment, the nucleotides can be modified nucleotides, unmodified nucleotides, or any combination thereof. The nucleotides can be ribonucleotides, deoxyribonucleotides, or any combination thereof. The siNA can contain at least five nucleotides. The siNA molecules described herein can contain modified nucleotides selected from 2'-O-methyl nucleotides and 2'-fluoro nucleotides.

[0219] In any embodiment, the first nucleotide sequence can comprise the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314, or 315. In any embodiment, the second nucleotide sequence can comprise the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, or 288-313.

[0220] In any embodiment, the siNA may reduce or inhibit the production of hydroxysteroid dehydrogenase. In any embodiment, the siNA may target the hydroxysteroid 17-beta dehydrogenase 13 (HSD17B13) gene.

[0221] In any embodiment, the siNA molecules described herein can comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more phosphorothioate internucleoside linkages. In any embodiment, the siNA molecules described herein can comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more mesyl phosphoramidate internucleoside linkage(s).

[0222] In any embodiment, the siNA molecules described herein may comprise a phosphorylation blocker. In any embodiment, the siNA molecules described herein may comprise a 5' stabilized end cap. In any embodiment, the siNA molecules described herein may comprise a galactosamine. In any embodiment, the siNA molecules described herein may comprise a conjugate moiety. In any embodiment, the siNA molecules described herein may comprise a destabilizing nucleotide. In any embodiment, the siNA molecules described herein may comprise a modified nucleotide. In any embodiment, the siNA molecules described herein may comprise a thermodestabilizing nucleotide.

[0223] In any embodiment, the siNA molecules described herein can comprise one or more blunt ends. In any embodiment, the siNA molecules described herein can comprise one or more overhangs.

[0224] In one embodiment, the siNA molecule comprises: (a) a sense strand comprising a first nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to an RNA corresponding to a target gene, wherein the first nucleotide sequence (i) is 15-30 nucleotides in length; and (ii) comprises 15 or more modified nucleotides independently selected from 2'-O-methyl nucleotides and 2'-fluoro nucleotides, wherein at least one modified nucleotide is a 2'-O-methyl nucleotide, and the nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of the first nucleotide sequence. and (b) an antisense strand comprising a second nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% complementary to RNA corresponding to the target gene, wherein the second nucleotide sequence (i) is 15 to 30 nucleotides in length; and (ii) comprises 15 or more modified nucleotides independently selected from 2'-O-methyl nucleotides and 2'-fluoro nucleotides, wherein at least one modified nucleotide is a 2'-O-methyl nucleotide and at least one modified nucleotide is a 2'-fluoro nucleotide.

[0225] In another embodiment, the present technology also provides a compound of formula (VIII): [ka] wherein the top strand is a sense strand comprising a first nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to an RNA corresponding to a target gene (wherein the first nucleotide sequence comprises 15-30 nucleotides); the bottom strand is an antisense strand comprising a second nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% complementary to an RNA corresponding to a target gene (wherein the second nucleotide sequence comprises 15-30 nucleotides); each A is independently a 2'-O-methyl nucleotide, or a nucleotide comprising a 5'-stabilized end cap or a phosphorylation blocker; B is a 2'-fluoro nucleotide; C represents an overhanging nucleotide and is a 2'-O-methyl nucleotide, a deoxynucleotide, or uracil; 1 = 1-6 nucleotides in length; each n 2 , n 6 , n 8 , q 3 , q 5 , q 7 , q 9 , q 11 and q 12 are independently 0-1 nucleotides in length; each n 3 and n 4 are independently 1-3 nucleotides in length; n 5 is 1-10 nucleotides in length; n 7 is 0-4 nucleotides in length; each n 9 , q 1 and q 2 are independently 0-2 nucleotides in length; q 4 is 0-3 nucleotides in length; q 6 is 0-5 nucleotides in length; q 8 is 2-7 nucleotides in length; and q 10 are 2-11 nucleotides in length.

[0226] An exemplary siNA molecule of the present disclosure is shown in FIG. 1. As shown in FIG. 1, the exemplary siNA molecule includes a sense strand (101) and an antisense strand (102). The sense strand (101) may include a first oligonucleotide sequence (103). The first oligonucleotide sequence (103) may include one or more phosphorothioate internucleoside linkages (109). The phosphorothioate internucleoside linkages (109) may be present between nucleotides at the 5' or 3' end of the first oligonucleotide sequence (103). The phosphorothioate internucleoside linkages (109) may be present between the first three nucleotides from the 5' end of the first oligonucleotide sequence (103). The first oligonucleotide sequence (103) may include one or more 2'-fluoro nucleotides (110). The first oligonucleotide sequence (103) may include one or more 2'-O-methyl nucleotides (111). The first oligonucleotide sequence (103) may contain 15 or more modified nucleotides independently selected from 2'-fluoronucleotides (110) and 2'-O-methyl nucleotides (111). The sense strand (101) may further contain a phosphorylation blocker (105). The sense strand (101) may further contain a galactosamine (106). The antisense strand (102) may contain a second oligonucleotide sequence (104). The second oligonucleotide sequence (104) may contain one or more phosphorothioate internucleoside linkages (109). The phosphorothioate internucleoside linkages (109) may be present between nucleotides at the 5' or 3' end of the second oligonucleotide sequence (104). The phosphorothioate internucleoside linkages (109) may be present between the first three nucleotides from the 5' end of the second oligonucleotide sequence (104). A phosphorothioate internucleoside linkage (109) may be present between the first three nucleotides from the 3' end of the second oligonucleotide sequence (104). The second oligonucleotide sequence (104) may contain one or more 2'-fluoro nucleotides (110). The second oligonucleotide sequence (104) may contain one or more 2'-O-methyl nucleotides (111).The second oligonucleotide sequence (104) may contain 15 or more modified nucleotides independently selected from 2'-fluoronucleotides (110) and 2'-O-methyl nucleotides (111). The antisense strand (102) may further contain a 5'-stabilized end cap (107). The siNA may further contain one or more blunt ends. Alternatively or additionally, one end of the siNA may contain an overhang (108). The overhang (108) may be part of the sense strand (101). The overhang (108) may be part of the antisense strand (102). The overhang (108) may be different from the first nucleotide sequence (103). The overhang (108) may be different from the second nucleotide sequence (104). The overhang (108) may be part of the first nucleotide sequence (103). The overhang (108) may be part of the second nucleotide sequence (104). The overhang (108) may comprise one or more nucleotides. The overhang (108) may comprise one or more deoxyribonucleotides. The overhang (108) may comprise one or more modified nucleotides. The overhang (108) may comprise one or more modified ribonucleotides. The sense strand (101) may be shorter than the antisense strand (102). The sense strand (101) may be the same length as the antisense strand (102). The sense strand (101) may be longer than the antisense strand (102).

[0227] An exemplary siNA molecule of the present disclosure is shown in Figure 2. As shown in Figure 2, the exemplary siNA molecule includes a sense strand (201) and an antisense strand (202). The sense strand (201) can include a first oligonucleotide sequence (203). The first oligonucleotide sequence (203) can include one or more phosphorothioate internucleoside linkages (209). The phosphorothioate internucleoside linkages (209) can be present between nucleotides at the 5' or 3' end of the first oligonucleotide sequence (203). The phosphorothioate internucleoside linkages (209) can be present between the first three nucleotides from the 5' end of the first oligonucleotide sequence (203). The first oligonucleotide sequence (203) can include one or more 2'-fluoronucleotides (210). The first oligonucleotide sequence (203) may contain one or more 2'-O-methyl nucleotides (211). The first oligonucleotide sequence (203) may contain 15 or more modified nucleotides independently selected from 2'-fluoro nucleotides (210) and 2'-O-methyl nucleotides (211). The sense strand (201) may further contain a phosphorylation blocker (205). The sense strand (201) may further contain a galactosamine (206). The antisense strand (202) may contain a second oligonucleotide sequence (204). The second oligonucleotide sequence (204) may contain one or more phosphorothioate internucleoside linkages (209). The phosphorothioate internucleoside linkages (209) may be present between nucleotides at the 5' or 3' end of the second oligonucleotide sequence (204). The phosphorothioate internucleoside linkage (209) may be present between the first three nucleotides from the 5' end of the second oligonucleotide sequence (204). The phosphorothioate internucleoside linkage (209) may be present between the first three nucleotides from the 3' end of the second oligonucleotide sequence (204). The second oligonucleotide sequence (204) may contain one or more 2'-fluoro nucleotides (210). The second oligonucleotide sequence (204) may contain one or more 2'-O-methyl nucleotides (211).The second oligonucleotide sequence (204) may comprise 15 or more modified nucleotides independently selected from 2'-fluoro nucleotides (210) and 2'-O-methyl nucleotides (211). The antisense strand (202) may further comprise a 5'-stabilized endcap (207). The siNA may further comprise one or more overhangs (208). The overhang (208) may be part of the sense strand (201). The overhang (208) may be part of the antisense strand (202). The overhang (208) may be different from the first nucleotide sequence (203). The overhang (208) may be different from the second nucleotide sequence (204). The overhang (208) may be part of the first nucleotide sequence (203). The overhang (208) may be part of the second nucleotide sequence (204). The overhang (208) may be adjacent to the 3' end of the first nucleotide sequence (203). The overhang (208) may be adjacent to the 5' end of the first nucleotide sequence (203). The overhang (208) may be adjacent to the 3' end of the second nucleotide sequence (204). The overhang (208) may be adjacent to the 5' end of the second nucleotide sequence (204). The overhang (208) may comprise one or more nucleotides. The overhang (208) may comprise one or more deoxyribonucleotides. The overhang (208) may comprise a TT sequence. The overhang (208) may comprise one or more modified nucleotides. The overhang (208) may comprise one or more modified nucleotides disclosed herein (e.g., 2-fluoro nucleotides, 2'-O-methyl nucleotides, 2'-fluoro nucleotide mimics, 2'-O-methyl nucleotide mimics, or nucleotides containing modified nucleobases). The overhang (208) may comprise one or more modified ribonucleotides. The sense strand (201) may be shorter than the antisense strand (202). The sense strand (201) may be the same length as the antisense strand (202). The sense strand (201) may be longer than the antisense strand (202).

[0228] 3A-3H show exemplary ds-siNA modification patterns. As shown in FIG. 3A-3H, exemplary ds-siNA molecules have the following formula: [ka] wherein the top strand is a sense strand comprising a first nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to an RNA corresponding to a target gene (wherein the first nucleotide sequence comprises 15-30 nucleotides); the bottom strand is an antisense strand comprising a second nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% complementary to an RNA corresponding to a target gene (wherein the second nucleotide sequence comprises 15-30 nucleotides); each A is independently a 2'-O-methyl nucleotide, or a nucleotide comprising a 5'-stabilized end cap or a phosphorylation blocker; B is a 2'-fluoro nucleotide; C represents an overhanging nucleotide and is a 2'-O-methyl nucleotide, a deoxynucleotide, or uracil; 1 = 1-6 nucleotides in length; each n 2 , n 6 , n 8 , q 3 , q 5 , q 7 , q 9 , q 11 and q 12 are independently 0-1 nucleotides in length; each n 3 and n 4 are independently 1-3 nucleotides in length; n 5 is 1-10 nucleotides in length; n 7 is 0-4 nucleotides in length; each n 9 , q 1 and q 2 are independently 0-2 nucleotides in length; q 4 is 0-3 nucleotides in length; q 6 is 0-5 nucleotides in length; q8 is 2-7 nucleotides in length; and q 10 are 2-11 nucleotides in length.

[0229] The ds-siNA may further comprise a conjugate moiety. The conjugate moiety may comprise any of the galactosamines disclosed herein. The ds-siNA may further comprise (i) phosphorothioate internucleoside linkages between nucleotides 1 and 2 and 2 and 3 from the 5' end of the sense strand; and (ii) phosphorothioate internucleoside linkages between nucleotides 1 and 2, 2 and 3, 19 and 20, and 20 and 21 from the 5' end of the antisense strand. The ds-siNA may further comprise a 5'-stabilizing endcap. The 5'-stabilizing endcap may be vinyl phosphonate. The 5'-stabilizing endcap may be attached to the 5' end of the antisense strand. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a 5'-stabilizing endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a 5'-stabilized endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is further modified to include a phosphorylation blocker.

[0230] An exemplary ds-siNA molecule has the formula: [ka] wherein the top strand is a sense strand comprising a first nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to an RNA corresponding to a target gene (wherein the first nucleotide sequence comprises 15-30 nucleotides); the bottom strand is an antisense strand comprising a second nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% complementary to an RNA corresponding to a target gene (wherein the second nucleotide sequence comprises 15-30 nucleotides); each A is independently a 2'-O-methyl nucleotide, or a nucleotide comprising a 5'-stabilized end cap or a phosphorylation blocker; B is a 2'-fluoro nucleotide; and C represents an overhanging nucleotide and is a 2'-O-methyl nucleotide, a deoxynucleotide, or uracil.

[0231] The ds-siNA may further comprise a conjugate moiety. The conjugate moiety may comprise any of the galactosamines disclosed herein. The ds-siNA may further comprise (i) phosphorothioate internucleoside linkages between nucleotides 1 and 2 and 2 and 3 from the 5' end of the sense strand; and (ii) phosphorothioate internucleoside linkages between nucleotides 1 and 2, 2 and 3, 19 and 20, and 20 and 21 from the 5' end of the antisense strand. The ds-siNA may further comprise a 5'-stabilizing endcap. The 5'-stabilizing endcap may be a vinyl phosphonate. The vinyl phosphonate may be a deuterated vinyl phosphonate. The deuterated vinyl phosphonate may be a mono-deuterated vinyl phosphonate. The deuterated vinyl phosphonate may be a mono-di-deuterated vinyl phosphonate. The 5'-stabilized endcap may be attached to the 5'-end of the antisense strand. The 5'-stabilized endcap may be attached to the 3'-end of the antisense strand. The 5'-stabilized endcap may be attached to the 5'-end of the sense strand. The 5'-stabilized endcap may be attached to the 3'-end of the sense strand. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5'-end of the sense strand is further modified to include a 5'-stabilized endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5'-end of the antisense strand is further modified to include a 5'-stabilized endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5'-end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3'-end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5'-end of the antisense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is further modified to include a phosphorylation blocker.

[0232] Exemplary ds-siNAs shown in Figures 3A-3H include (i) a sense strand containing 19-21 nucleotides; and (ii) an antisense strand containing 21-23 nucleotides. The ds-siNA can further include (iii) a conjugate moiety, wherein the conjugate moiety is attached to the 3'-end of the antisense strand. The ds-siNA can include a two-nucleotide overhang consisting of nucleotides 20 and 21 from the 5'-end of the antisense strand. The ds-siNA can include a two-nucleotide overhang consisting of nucleotides 22 and 23 from the 5'-end of the antisense strand. The ds-siNA can further include one, two, three, four, five, or six or more phosphorothioate (ps) internucleoside linkages. At least one phosphorothioate internucleoside linkage can be present between nucleotides 1 and 2 or 2 and 3 from the 5'-end of the sense strand. At least one phosphorothioate internucleoside linkage may be present between nucleotides 1 and 2 or 2 and 3 from the 5' end of the antisense strand. At least one phosphorothioate internucleoside linkage may be present between nucleotides 19 and 20, 20 and 21, 21 and 22, or 22 and 23 from the 5' end of the antisense strand. As shown in Figures 3A-3H, nucleotides 4-6 in the sense strand may be 2'-fluoro nucleotides. As shown in Figures 3A-3H, nucleotides 2-5 in the antisense strand may be 2'-fluoro nucleotides. As shown in Figures 3A-3H, nucleotides 13-15 in the sense strand may be 2'-O-methyl nucleotides. As shown in Figures 3A-3H, nucleotides 14-19 in the antisense strand may be 2'-O-methyl nucleotides. As shown in Figures 3A-3H, the ds-siNA does not contain base pairs between the 2'-fluoro nucleotides of the sense and antisense strands. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to contain a 5' stabilized end cap.In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a 5'-stabilized endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is further modified to include a phosphorylation blocker.

[0233] In some embodiments, (a) the sense strand may comprise a first nucleotide sequence consisting of 17-23 nucleotides, wherein 2'-fluoro nucleotides are present at positions 3, 7-9, 12, and 17 from the 5' end of the first nucleotide sequence, and wherein 2'-O-methyl nucleotides are present at positions 1, 2, 4-6, 10, 11, and 13-16 from the 5' end of the first nucleotide sequence; and (b) the antisense strand may comprise a second nucleotide sequence consisting of 17-23 nucleotides. In some embodiments, the 2'-fluoro nucleotides are present at positions 2 and 14 from the 5' end of the second nucleotide sequence, and the 2'-O-methyl nucleotides are present at positions 1, 3-13, and 15-17 from the 5' end of the second nucleotide sequence. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:3 modification pattern, where one nucleotide is a 2'-fluoro nucleotide and three nucleotides are 2'-O-methyl nucleotides. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:2 modification pattern, where one nucleotide is a 2'-fluoro nucleotide and two nucleotides are 2'-O-methyl nucleotides. In any embodiment, the first nucleotide sequence consists of 19 nucleotides. In any embodiment, the 2'-O-methyl nucleotides are present at positions 18 and 19 from the 5' end of the first nucleotide sequence. In any embodiment, the second nucleotide sequence consists of 21 nucleotides. In any embodiment, the 2'-O-methyl nucleotides are present at positions 18-21 or 19-21 from the 5' end of the second nucleotide sequence.As shown in Figure 3A, the ds-siNA may comprise: (a) a sense strand consisting of 19 nucleotides, in which 2'-fluoro nucleotides are present at positions 3, 7-9, 12, and 17 from the 5' end of the sense strand, and 2'-O-methyl nucleotides are present at positions 1, 2, 4-6, 10, 11, 13-16, 18, and 19 from the 5' end of the sense strand; and (b) an antisense strand consisting of 21 nucleotides, in which nucleotides 2 and 14 from the 5' end of the antisense strand are 2'-fluoro nucleotides; and nucleotides 1, 3-13, and 15-21 are 2'-O-methyl nucleotides. The ds-siNA may further comprise a conjugate moiety attached to the 3' end of the sense strand. The ds-siNA may further comprise (i) phosphorothioate internucleoside linkages between nucleotides 1 and 2 and 2 and 3 from the 5' end of the sense strand; and (ii) phosphorothioate internucleoside linkages between nucleotides 1 and 2, 2 and 3, 19 and 20, and 20 and 21 from the 5' end of the antisense strand. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a 5'-stabilizing endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a 5'-stabilizing endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is further modified to include a phosphorylation blocker.In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides in the sense or antisense strand are 2'-fluoro nucleotide mimics. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides in the sense or antisense strand are fB, fN, f(4nh)Q, f4P, f2P, or fX nucleotides. In some embodiments, at least one, two, three, or four or more 2'-O-methyl nucleotides in the sense or antisense strand are 2'-O-methyl nucleotide mimics.In some embodiments, one or more nucleotides of the sense strand and / or antisense strand can be a 3',4'seco modified nucleotide (e.g., mun34) in which the bond between the 3' and 4' positions of the furanose ring is cleaved.

[0234] In some embodiments, (a) the sense strand comprises a first nucleotide sequence consisting of 17-23 nucleotides, wherein 2'-fluoro nucleotides are present at positions 3, 7, 8, and 17 from the 5' end of the first nucleotide sequence, and wherein 2'-O-methyl nucleotides are present at positions 1, 2, 4-6, and 9-16 from the 5' end of the first nucleotide sequence; and (b) the antisense strand comprises a second nucleotide sequence consisting of 17-23 nucleotides. In some embodiments, the 2'-fluoro nucleotides are present at positions 2 and 14 from the 5' end of the second nucleotide sequence, and the 2'-O-methyl nucleotides are present at positions 1, 3-13, and 15-17 from the 5' end of the second nucleotide sequence. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:3 modification pattern, wherein one nucleotide is a 2'-fluoro nucleotide and three nucleotides are 2'-O-methyl nucleotides. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:2 modification pattern, where one nucleotide is a 2'-fluoro nucleotide and two nucleotides are 2'-O-methyl nucleotides. In any embodiment, the first nucleotide sequence consists of 19 nucleotides. In any embodiment, the 2'-O-methyl nucleotides are present at positions 18 and 19 from the 5' end of the first nucleotide sequence. In any embodiment, the second nucleotide sequence consists of 21 nucleotides. In any embodiment, the 2'-O-methyl nucleotides are present at positions 18-21 or 19-21 from the 5' end of the second nucleotide sequence.As shown in Figure 3B, the ds-siNA may comprise: (a) a sense strand consisting of 19 nucleotides, in which 2'-fluoro nucleotides are present at positions 3, 7, 8, and 17 from the 5' end of the sense strand, and 2'-O-methyl nucleotides are present at positions 1, 2, 4-6, 9-16, 18, and 19 from the 5' end of the sense strand; and (b) an antisense strand consisting of 21 nucleotides, in which nucleotides 2 and 14 from the 5' end of the antisense strand are 2'-fluoro nucleotides; and nucleotides 1, 3-13, and 15-21 are 2'-O-methyl nucleotides. The ds-siNA may further comprise a conjugate moiety attached to the 3' end of the sense strand. The ds-siNA may further comprise (i) phosphorothioate internucleoside linkages between nucleotides 1 and 2 and 2 and 3 from the 5' end of the sense strand; and (ii) phosphorothioate internucleoside linkages between nucleotides 1 and 2, 2 and 3, 19 and 20, and 20 and 21 from the 5' end of the antisense strand. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a 5'-stabilizing endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a 5'-stabilizing endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is further modified to include a phosphorylation blocker.In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides in the sense or antisense strand are 2'-fluoro nucleotide mimics. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides in the sense or antisense strand are fB, fN, f(4nh)Q, f4P, f2P, or fX nucleotides. In some embodiments, at least one, two, three, or four or more 2'-O-methyl nucleotides in the sense or antisense strand are 2'-O-methyl nucleotide mimics.In some embodiments, one or more nucleotides of the sense strand and / or antisense strand can be a 3',4'seco modified nucleotide (e.g., mun34) in which the bond between the 3' and 4' positions of the furanose ring is cleaved.

[0235] In some embodiments, (a) the sense strand comprises a first nucleotide sequence consisting of 17-23 nucleotides, wherein 2'-fluoro nucleotides are located at positions 3, 7-9, and 17 from the 5' end of the first nucleotide sequence, and wherein 2'-O-methyl nucleotides are located at positions 1, 2, 4-6, and 10-16 from the 5' end of the first nucleotide sequence; and (b) the antisense strand comprises a second nucleotide sequence consisting of 17-23 nucleotides. In some embodiments, the 2'-fluoro nucleotides are located at positions 2, 6, 10, and 14 from the 5' end of the second nucleotide sequence, and the 2'-O-methyl nucleotides are located at positions 1, 3-5, 7-9, 11-13, and 15-17 from the 5' end of the second nucleotide sequence. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:3 modification pattern, where one nucleotide is a 2'-fluoro nucleotide and three nucleotides are 2'-O-methyl nucleotides. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:2 modification pattern, where one nucleotide is a 2'-fluoro nucleotide and two nucleotides are 2'-O-methyl nucleotides. In any embodiment, the first nucleotide sequence consists of 19 nucleotides. In any embodiment, the 2'-O-methyl nucleotides are present at positions 18 and 19 from the 5' end of the first nucleotide sequence. In any embodiment, the second nucleotide sequence consists of 21 nucleotides. In any embodiment, the 2'-O-methyl nucleotides are present at positions 19-21 from the 5' end of the second nucleotide sequence.As shown in Figure 3C, the ds-siNA may comprise: (a) a sense strand consisting of 19 nucleotides, in which 2'-fluoro nucleotides are present at positions 3, 7-9, 12, and 17 from the 5' end of the sense strand, and 2'-O-methyl nucleotides are present at positions 1, 2, 4-6, 10, 11, 13-16, 18, and 19 from the 5' end of the sense strand; and (b) an antisense strand consisting of 21 nucleotides, in which the nucleotides in the antisense strand comprise an alternating 1:3 modification pattern, and one nucleotide is a 2'-fluoro nucleotide and three nucleotides are 2'-O-methyl nucleotides. The ds-siNA may further comprise a conjugate moiety attached to the 3' end of the sense strand. The ds-siNA may further comprise (i) phosphorothioate internucleoside linkages between nucleotides 1 and 2 and 2 and 3 from the 5' end of the sense strand; and (ii) phosphorothioate internucleoside linkages between nucleotides 1 and 2, 2 and 3, 19 and 20, and 20 and 21 from the 5' end of the antisense strand. The ds-siNA may comprise a 2-5 alternating 1:3 modification pattern on the antisense strand. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a 5'-stabilizing endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a 5'-stabilizing endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is further modified to include a phosphorylation blocker.In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides in the sense or antisense strand are 2'-fluoro nucleotide mimics. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides on the sense strand are fB, fN, f(4nh)Q, f4P, f2P, or fX nucleotides. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides on the antisense strand are fB, fN, f(4nh)Q, f4P, f2P, or fX nucleotides.In some embodiments, at least one, two, three, or four or more 2'-O-methyl nucleotides in the sense or antisense strand are 2'-O-methyl nucleotide mimics. In some embodiments, one or more nucleotides in the sense and / or antisense strand can be 3',4' seco-modified nucleotides in which the bond between the 3' and 4' positions of the furanose ring is cleaved (e.g., mun34).

[0236] In some embodiments, (a) the sense strand comprises a first nucleotide sequence consisting of 17-23 nucleotides, wherein 2'-fluoro nucleotides are present at positions 5 and 7-9 from the 5' end of the first nucleotide sequence, and wherein 2'-O-methyl nucleotides are present at positions 1-4, 6, and 10-17 from the 5' end of the first nucleotide sequence; and (b) the antisense strand comprises a second nucleotide sequence consisting of 17-23 nucleotides. In some embodiments, the 2'-fluoro nucleotides are present at positions 2, 6, 10, and 14 from the 5' end of the second nucleotide sequence, and the 2'-O-methyl nucleotides are present at positions 1, 3-5, 7-9, 11-13, and 15-17 from the 5' end of the second nucleotide sequence. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:3 modification pattern, where one nucleotide is a 2'-fluoro nucleotide and three nucleotides are 2'-O-methyl nucleotides. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:2 modification pattern, where one nucleotide is a 2'-fluoro nucleotide and two nucleotides are 2'-O-methyl nucleotides. In any embodiment, the first nucleotide sequence consists of 19 nucleotides. In any embodiment, the 2'-O-methyl nucleotides are present at positions 18 and 19 from the 5' end of the first nucleotide sequence. In any embodiment, the second nucleotide sequence consists of 21 nucleotides. In any embodiment, the 2'-fluoro nucleotide is present at position 18 from the 5' end of the second nucleotide sequence. In any embodiment, the 2'-O-methyl nucleotide is present at positions 19-21 from the 5' end of the second nucleotide sequence.As shown in Figure 3D, the ds-siNA may comprise: (a) a sense strand consisting of 19 nucleotides, in which 2'-fluoro nucleotides are present at positions 5 and 7-9 from the 5' end of the sense strand, and 2'-O-methyl nucleotides are present at positions 1-4, 6, and 10-19 from the 5' end of the sense strand; and (b) an antisense strand consisting of 21 nucleotides, in which the nucleotides of the antisense strand comprise an alternating 1:3 modification pattern, in which one nucleotide is a 2'-fluoro nucleotide and three nucleotides are 2'-O-methyl nucleotides. The ds-siNA may further comprise a conjugate moiety attached to the 3' end of the sense strand. The ds-siNA may further comprise (i) phosphorothioate internucleoside linkages between nucleotides 1 and 2 and 2 and 3 from the 5' end of the sense strand; and (ii) phosphorothioate internucleoside linkages between nucleotides 1 and 2, 2 and 3, 19 and 20, and 20 and 21 from the 5' end of the antisense strand. The ds-siNA may comprise a 2-5 alternating 1:3 modification pattern on the antisense strand. The alternating 1:3 modification pattern may start at any of nucleotides 2, 6, 10, 14, and / or 18 from the 5' end of the antisense strand. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a 5'-stabilizing endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a 5'-stabilized endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is further modified to include a phosphorylation blocker.In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides in the sense or antisense strand are 2'-fluoro nucleotide mimics. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides on the sense strand are fB, fN, f(4nh)Q, f4P, or f2P nucleotides. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides on the antisense strand are fB, fN, f(4nh)Q, f4P, or f2P nucleotides.In some embodiments, at least one, two, three, or four or more 2'-O-methyl nucleotides in the sense or antisense strand are 2'-O-methyl nucleotide mimics. In some embodiments, one or more nucleotides in the sense and / or antisense strand can be 3',4' seco-modified nucleotides in which the bond between the 3' and 4' positions of the furanose ring is cleaved (e.g., mun34).

[0237] In some embodiments, (a) the sense strand comprises a first nucleotide sequence consisting of 17-23 nucleotides, wherein 2'-fluoro nucleotides are located at positions 5 and 7-9 from the 5' end of the first nucleotide sequence, and wherein 2'-O-methyl nucleotides are located at positions 1-4, 6, and 10-17 from the 5' end of the first nucleotide sequence; and (b) the antisense strand comprises a second nucleotide sequence consisting of 17-23 nucleotides. In some embodiments, the 2'-fluoro nucleotides are located at positions 2, 5, 8, 14, and 17 from the 5' end of the first nucleotide sequence, and the 2'-O-methyl nucleotides are located at positions 1, 3, 4, 6, 7, 9-13, 15, and 16 from the 5' end of the first nucleotide sequence. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:3 modification pattern, where one nucleotide is a 2'-fluoro nucleotide and three nucleotides are 2'-O-methyl nucleotides. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:2 modification pattern, where one nucleotide is a 2'-fluoro nucleotide and two nucleotides are 2'-O-methyl nucleotides. In any embodiment, the first nucleotide sequence consists of 19 nucleotides. In any embodiment, the 2'-O-methyl nucleotides are present at positions 18 and 19 from the 5' end of the first nucleotide sequence. In any embodiment, the second nucleotide sequence consists of 21 nucleotides. In any embodiment, the 2'-O-methyl nucleotides are present at positions 18-21 or 19-21 from the 5' end of the second nucleotide sequence.As shown in Figure 3E, the ds-siNA may comprise: (a) a sense strand consisting of 19 nucleotides, in which 2'-fluoro nucleotides are present at positions 5 and 7-9 from the 5' end of the sense strand, and 2'-O-methyl nucleotides are present at positions 1-4, 6, and 10-19 from the 5' end of the sense strand; and (b) an antisense strand consisting of 21 nucleotides, in which the nucleotides of the antisense strand comprise an alternating 1:2 modification pattern, in which one nucleotide is a 2'-fluoro nucleotide and two nucleotides are 2'-O-methyl nucleotides. The ds-siNA may further comprise a conjugate moiety attached to the 3' end of the sense strand. The ds-siNA may further comprise (i) phosphorothioate internucleoside linkages between nucleotides 1 and 2 and 2 and 3 from the 5' end of the sense strand; and (ii) phosphorothioate internucleoside linkages between nucleotides 1 and 2, 2 and 3, 19 and 20, and 20 and 21 from the 5' end of the antisense strand. The ds-siNA may comprise 2-5 alternating 1:2 modification patterns on the antisense strand. The alternating 1:2 modification patterns may start at any of nucleotides 2, 5, 8, 14, and / or 17 from the 5' end of the antisense strand. In some embodiments, the ds-siNA comprises: (a) a 19-nucleotide sense strand, in which 2'-fluoro nucleotides are located at positions 5 and 7-9 from the 5' end of the sense strand, and 2'-O-methyl nucleotides are located at positions 1-4, 6, and 10-19 from the 5' end of the sense strand; and (b) a 21-nucleotide antisense strand, in which 2'-fluoro nucleotides are located at positions 2, 5, 8, 14, and 17 from the 5' end of the antisense strand, and 2'-O-methyl nucleotides are located at positions 1, 3, 4, 6, 7, 9-13, 15, 16, and 18-21 from the 5' end of the sense strand. In some embodiments, the 2'-O-methyl nucleotide located at position 1 from the 5' end of the sense strand is further modified to include a 5'-stabilizing endcap.In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a 5'-stabilized endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide.In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides in the sense strand or the antisense strand are 2'-fluoro nucleotide mimics. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides on the sense strand are fB, fN, f(4nh)Q, f4P, or f2P nucleotides. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides on the antisense strand are fB, fN, f(4nh)Q, f4P, or f2P nucleotides. In some embodiments, at least one, two, three, or four or more 2'-O-methyl nucleotides in the sense or antisense strand are 2'-O-methyl nucleotide mimics. In some embodiments, one or more nucleotides in the sense and / or antisense strand can be 3',4' seco-modified nucleotides in which the bond between the 3' and 4' positions of the furanose ring is cleaved (e.g., mun34).

[0238] In some embodiments, (a) the sense strand comprises a first nucleotide sequence consisting of 17-23 nucleotides, wherein 2'-fluoro nucleotides are located at positions 5 and 7-9 from the 5' end of the first nucleotide sequence, and wherein 2'-O-methyl nucleotides are located at positions 1-4, 6, and 10-17 from the 5' end of the first nucleotide sequence; and (b) the antisense strand comprises a second nucleotide sequence consisting of 17-23 nucleotides. In some embodiments, the 2'-fluoro nucleotides are located at positions 2, 6, 14, and 16 from the 5' end of the second nucleotide sequence, and the 2'-O-methyl nucleotides are located at positions 1, 3-5, 7-13, 15, and 17 from the 5' end of the second nucleotide sequence. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:3 modification pattern, where one nucleotide is a 2'-fluoro nucleotide and three nucleotides are 2'-O-methyl nucleotides. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:2 modification pattern, where one nucleotide is a 2'-fluoro nucleotide and two nucleotides are 2'-O-methyl nucleotides. In any embodiment, the first nucleotide sequence consists of 19 nucleotides. In any embodiment, the 2'-O-methyl nucleotides are present at positions 18 and 19 from the 5' end of the first nucleotide sequence. In any embodiment, the second nucleotide sequence consists of 21 nucleotides. In any embodiment, the 2'-O-methyl nucleotides are present at positions 18-21 or 19-21 from the 5' end of the second nucleotide sequence.As shown in Figure 3F, the ds-siNA may comprise: (a) a 19-nucleotide sense strand (wherein 2'-fluoro nucleotides are located at positions 5 and 7-9 from the 5' end of the sense strand, and 2'-O-methyl nucleotides are located at positions 1-4, 6, and 10-19 from the 5' end of the sense strand); and (b) a 21-nucleotide antisense strand (wherein 2'-fluoro nucleotides are located at positions 2, 6, 14, and 16 from the 5' end of the antisense strand, and 2'-O-methyl nucleotides are located at positions 1, 3-5, 7-13, 15, and 17-21 from the 5' end of the antisense strand). The ds-siNA may further comprise a conjugate moiety attached to the 3' end of the sense strand. The ds-siNA may further comprise (i) phosphorothioate internucleoside linkages between nucleotides 1 and 2 and 2 and 3 from the 5' end of the sense strand; and (ii) phosphorothioate internucleoside linkages between nucleotides 1 and 2, 2 and 3, 19 and 20, and 20 and 21 from the 5' end of the antisense strand. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides in the sense strand or antisense strand are fB, fN, f(4nh)Q, f4P, or f2P nucleotides. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides in the sense strand or antisense strand are f4P nucleotides. In some embodiments, at least one, two, three, or four of the 2'-fluoro nucleotides at positions 2, 6, 14, and 16 from the 5' end of the antisense strand are f4P nucleotides. In some embodiments, at least one of the 2'-fluoro nucleotides at positions 2, 6, 14, and 16 from the 5' end of the antisense strand is an f4P nucleotide. In some embodiments, at least two of the 2'-fluoro nucleotides at positions 2, 6, 14, and 16 from the 5' end of the antisense strand are f4P nucleotides. In some embodiments, no more than three of the 2'-fluoro nucleotides at positions 2, 6, 14, and 16 from the 5' end of the antisense strand are f4P nucleotides.In some embodiments, no more than two of the 2'-fluoro nucleotides at positions 2, 6, 14, and 16 from the 5' end of the antisense strand are f4P nucleotides. In some embodiments, the 2'-fluoro nucleotide at position 2 from the 5' end of the antisense strand is an f4P nucleotide. In some embodiments, the 2'-fluoro nucleotide at position 6 from the 5' end of the antisense strand is an f4P nucleotide. In some embodiments, the 2'-fluoro nucleotide at position 14 from the 5' end of the antisense strand is an f4P nucleotide. In some embodiments, the 2'-fluoro nucleotide at position 16 from the 5' end of the antisense strand is an f4P nucleotide. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides in the sense or antisense strand are f2P nucleotides. In some embodiments, at least one, two, three, or four of the 2'-fluoro nucleotides at positions 2, 6, 14, and 16 from the 5' end of the antisense strand are f2P nucleotides. In some embodiments, at least one of the 2'-fluoro nucleotides at positions 2, 6, 14, and 16 from the 5' end of the antisense strand is an f2P nucleotide. In some embodiments, at least two of the 2'-fluoro nucleotides at positions 2, 6, 14, and 16 from the 5' end of the antisense strand are f2P nucleotides. In some embodiments, no more than three of the 2'-fluoro nucleotides at positions 2, 6, 14, and 16 from the 5' end of the antisense strand are f2P nucleotides. In some embodiments, no more than two of the 2'-fluoro nucleotides at positions 2, 6, 14, and 16 from the 5' end of the antisense strand are f2P nucleotides. In some embodiments, the 2'-fluoro nucleotide at position 2 from the 5' end of the antisense strand is an f2P nucleotide. In some embodiments, the 2'-fluoro nucleotide at position 6 from the 5' end of the antisense strand is an f2P nucleotide. In some embodiments, the 2'-fluoro nucleotide at position 14 from the 5' end of the antisense strand is an f2P nucleotide. In some embodiments, the 2'-fluoro nucleotide at position 16 from the 5' end of the antisense strand is an f2P nucleotide.In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a 5'-stabilizing endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a 5'-stabilizing endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide.In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides in the sense strand or the antisense strand are 2'-fluoro nucleotide mimics. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides on the sense strand are fB, fN, f(4nh)Q, f4P, or f2P nucleotides. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides on the antisense strand are fB, fN, f(4nh)Q, f4P, or f2P nucleotides. In some embodiments, at least one, two, three, or four or more 2'-O-methyl nucleotides in the sense or antisense strand are 2'-O-methyl nucleotide mimics. In some embodiments, one or more nucleotides in the sense and / or antisense strand can be 3',4' seco-modified nucleotides in which the bond between the 3' and 4' positions of the furanose ring is cleaved (e.g., mun34).

[0239] In some embodiments, (a) the sense strand comprises a first nucleotide sequence consisting of 17-23 nucleotides, wherein 2'-fluoro nucleotides are located at positions 5, 9-11, and 14 from the 5' end of the first nucleotide sequence, and wherein 2'-O-methyl nucleotides are located at positions 1-4, 6-8, 12, 13, and 15-17 from the 5' end of the first nucleotide sequence; and (b) the antisense strand comprises a second nucleotide sequence consisting of 17-23 nucleotides. In some embodiments, the 2'-fluoro nucleotides are located at positions 2 and 14 from the 5' end of the second nucleotide sequence, and the 2'-O-methyl nucleotides are located at positions 1, 3-13, and 15-17 from the 5' end of the second nucleotide sequence. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:3 modification pattern, where one nucleotide is a 2'-fluoro nucleotide and three nucleotides are 2'-O-methyl nucleotides. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:2 modification pattern, where one nucleotide is a 2'-fluoro nucleotide and two nucleotides are 2'-O-methyl nucleotides. In any embodiment, the first nucleotide sequence consists of 21 nucleotides. In any embodiment, the 2'-O-methyl nucleotides are present at positions 18, 20, and 21 from the 5' end of the first nucleotide sequence. In any embodiment, the 2'-fluoro nucleotide is present at position 19 from the 5' end of the first nucleotide sequence. In any embodiment, the second nucleotide sequence consists of 23 nucleotides. In any embodiment, the 2'-O-methyl nucleotides are present at positions 18-23 from the 5' end of the second nucleotide sequence.As shown in Figure 3G, the ds-siNA may comprise: (a) a 21-nucleotide sense strand (wherein 2'-fluoro nucleotides are located at positions 5, 9-11, 14, and 19 from the 5' end of the sense strand, and 2'-O-methyl nucleotides are located at positions 1-4, 6-8, 12, 13, 15-18, 20, and 21 from the 5' end of the sense strand); and (b) a 23-nucleotide antisense strand (wherein 2'-fluoro nucleotides are located at positions 2 and 14 from the 5' end of the antisense strand, and 2'-O-methyl nucleotides are located at positions 1, 3-13, and 15-23 from the 5' end of the antisense strand). The ds-siNA may further comprise a conjugate moiety attached to the 3' end of the sense strand. The ds-siNA may further comprise (i) phosphorothioate internucleoside linkages between nucleotides 1 and 2 and 2 and 3 from the 5' end of the sense strand; and (ii) phosphorothioate internucleoside linkages between nucleotides 1 and 2, 2 and 3, 19 and 20, and 20 and 21 from the 5' end of the antisense strand. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a 5'-stabilizing endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a 5'-stabilizing endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is further modified to include a phosphorylation blocker.In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides in the sense or antisense strand are 2'-fluoro nucleotide mimics. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides on the sense strand are fB, fN, f(4nh)Q, f4P, or f2P nucleotides. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides on the antisense strand are fB, fN, f(4nh)Q, f4P, or f2P nucleotides.In some embodiments, at least one, two, three, or four or more 2'-O-methyl nucleotides in the sense or antisense strand are 2'-O-methyl nucleotide mimics. In some embodiments, one or more nucleotides in the sense and / or antisense strand can be 3',4' seco-modified nucleotides in which the bond between the 3' and 4' positions of the furanose ring is cleaved (e.g., mun34).

[0240] In some embodiments, (a) the sense strand comprises a first nucleotide sequence consisting of 17-23 nucleotides, wherein 2'-fluoro nucleotides are located at positions 7 and 9-11 from the 5' end of the first nucleotide sequence, and wherein 2'-O-methyl nucleotides are located at positions 1-6, 8, and 12-17 from the 5' end of the first nucleotide sequence; and (b) the antisense strand comprises a second nucleotide sequence consisting of 17-23 nucleotides. In some embodiments, the 2'-fluoro nucleotides are located at positions 2, 6, 14, and 16 from the 5' end of the second nucleotide sequence, and the 2'-O-methyl nucleotides are located at positions 1, 3-5, 7-13, 15, and 17 from the 5' end of the second nucleotide sequence. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:3 modification pattern, where one nucleotide is a 2'-fluoro nucleotide and three nucleotides are 2'-O-methyl nucleotides. In some embodiments, the nucleotides in the second nucleotide sequence are arranged in an alternating 1:2 modification pattern, where one nucleotide is a 2'-fluoro nucleotide and two nucleotides are 2'-O-methyl nucleotides. In any embodiment, the first nucleotide sequence consists of 21 nucleotides. In any embodiment, the 2'-O-methyl nucleotides are located at positions 18-21 from the 5' end of the first nucleotide sequence. In any embodiment, the second nucleotide sequence consists of 23 nucleotides. In any embodiment, the 2'-O-methyl nucleotides are located at positions 18-21 from the 5' end of the second nucleotide sequence.As shown in Figure 3H, the ds-siNA may comprise: (a) a 21-nucleotide sense strand (wherein 2'-fluoro nucleotides are located at positions 7 and 9-11 from the 5' end of the sense strand, and 2'-O-methyl nucleotides are located at positions 1-6, 8, and 12-21 from the 5' end of the sense strand); and (b) a 23-nucleotide antisense strand (wherein 2'-fluoro nucleotides are located at positions 2, 6, 14, and 16 from the 5' end of the antisense strand, and 2'-O-methyl nucleotides are located at positions 1, 3-5, 7-13, 15, and 17-23 from the 5' end of the antisense strand). Optionally, the 22nd and 23rd nucleotides from the 5' end of the antisense strand may be unlocked nucleotides. Optionally, the ds-siNA may further comprise a conjugate moiety attached to the 3' end of the sense strand (not shown). The ds-siNA may comprise a stabilized endcap attached to the 5'-end of the antisense strand (as shown). The ds-siNA may further comprise (i) phosphorothioate internucleoside linkages between nucleotides 1 and 2, 2 and 3, and 20 and 21 from the 5'-end of the sense strand; and (ii) phosphorothioate internucleoside linkages between nucleotides 1 and 2, 2 and 3, 21 and 22, and 22 and 23 from the 5'-end of the antisense strand. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5'-end of the sense strand is further modified to comprise a 5'-stabilized endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5'-end of the antisense strand is further modified to comprise a 5'-stabilized endcap. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5'-end of the sense strand is further modified to comprise a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is further modified to include a phosphorylation blocker.In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is further modified to include a phosphorylation blocker. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide, a d2vd3U nucleotide, an omeco-d3U nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 5' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the sense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, the 2'-O-methyl nucleotide at position 1 from the 3' end of the antisense strand is a d2vd3 nucleotide, a d2vd3U nucleotide, an omeco-d3 nucleotide, an omeco-d3U nucleotide, a 4h nucleotide, a 4hU nucleotide, a v-mun nucleotide, a c2o-4h nucleotide, an omeco-munb nucleotide, a d2vm nucleotide, or a d2vmA nucleotide. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides in the sense or antisense strand are 2'-fluoro nucleotide mimics.In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides on the sense strand are fB, fN, f(4nh)Q, f4P, or f2P nucleotides. In some embodiments, at least one, two, three, or four or more 2'-fluoro nucleotides on the antisense strand are fB, fN, f(4nh)Q, f4P, or f2P nucleotides. In some embodiments, at least one, two, three, or four or more 2'-O-methyl nucleotides in the sense strand or antisense strand are 2'-O-methyl nucleotide mimics. In some embodiments, one or more nucleotides in the sense strand and / or antisense strand can be 3',4'seco-modified nucleotides (e.g., mun34) in which the bond between the 3' and 4' positions of the furanose ring is cleaved.

[0241] Any of the siNAs disclosed herein can comprise a sense strand and an antisense strand. The sense strand can comprise a first nucleotide sequence that is 15-30 nucleotides in length. The antisense strand can comprise a second nucleotide sequence that is 15-30 nucleotides in length.

[0242] In some embodiments, the double-stranded short interfering nucleic acid (ds-siNA) molecule comprises: (a) a sense strand (wherein at least one modified nucleotide is a 2'-O-methyl nucleotide and the nucleotide at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide): In some embodiments, the double-stranded short interfering nucleic acid (ds-siNA) molecule comprises: (a) a sense strand (wherein at least one modified nucleotide is a 2'-O-methyl nucleotide and the nucleotide at position 7 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide): In some embodiments, the double-stranded short interfering nucleic acid (ds-siNA) molecule comprises: (a) a sense strand (wherein at least one modified nucleotide is a 2'-O-methyl nucleotide and the nucleotides at positions 7, 9, 10 and / or 11 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides).

[0243] In some embodiments, the double-stranded short interfering nucleic acid (ds-siNA) molecule comprises (b) an antisense strand (wherein at least one modified nucleotide is a 2'-O-methyl nucleotide and the nucleotide at position 2, 5, 6, 8, 10, 14, 16, 17, and / or 18 from the 5' end of the second nucleotide sequence is a 2'-fluoro nucleotide): In some embodiments, the double-stranded short interfering nucleic acid (ds-siNA) molecule comprises (b) an antisense strand (wherein at least one modified nucleotide is a 2'-O-methyl nucleotide and the nucleotide at position 2 of the second nucleotide sequence is a 2'-fluoro nucleotide):

[0244] In some embodiments, the ds-siNA molecule comprises one or more phosphorothioate internucleoside linkages. In some embodiments, the ds-siNA molecule comprises one or more mesyl phosphoramidate internucleoside linkages. In some embodiments, the ds-siNA molecule may further comprise a phosphorylation blocker, galactosamine, a 5'-stabilizing endcap, a conjugate moiety, a destabilizing nucleotide, a modified nucleotide, a thermally destabilizing nucleotide, or a combination of two or more thereof. In some embodiments, the sense strand further comprises a phosphorylation blocker or galactosamine. In some embodiments, the antisense strand further comprises a 5'-stabilizing endcap. In some embodiments, the sense strand further comprises a phosphorylation blocker or galactosamine, and the antisense strand further comprises a 5'-stabilizing endcap.

[0245] The present technology provides compositions comprising one or more of the siNA molecules described herein. The present technology further provides compositions comprising two or more of the siNA molecules described herein.

[0246] The present technology provides compositions comprising any of the described siNA molecules and a pharmaceutically acceptable carrier or diluent.

[0247] The present technology provides compositions comprising two or more of the siNA molecules described herein for use as pharmaceuticals.

[0248] The present technology provides compositions comprising any of the described siNA molecules and a pharmaceutically acceptable carrier or diluent for use as a pharmaceutical.

[0249] The present technology provides a method of treating a disease in a subject in need thereof, wherein the method comprises administering to the subject any of the siNA molecules described herein.

[0250] The present technology provides for the use of any of the siNA molecules described herein in the manufacture of a medicament for treating a disease.

[0251] Short interfering nucleic acid (siNA) molecules As noted above, the present disclosure provides siNA molecules containing modified nucleotides. Any of the siNA molecules described herein may be double-stranded siNA (ds-siNA) molecules. The terms "siNA molecule" and "ds-siNA molecule" may be used interchangeably. In some embodiments, the ds-siNA molecule comprises a sense strand and an antisense strand. The siNA comprises any of the first nucleotide sequence, second nucleotide sequence, sense strand sequence, or antisense strand sequence disclosed herein. The siNA may comprise 5 to 100, 5 to 90, 10 to 100, 10 to 90, 10 to 80, 10 to 70, 10 to 60, 10 to 50, 10 to 30, 10 to 25, 15 to 100, 15 to 90, 15 to 80, 15 to 70, 15 to 60, 15 to 50, 15 to 30, or 15 to 25 nucleotides. The siNA may contain at least 5, 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, or 40 nucleotides. The siNA may contain no more than 50, 45, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, or 19 nucleotides. The nucleotides may be modified nucleotides. The siNA may be single-stranded. The siNA may be double-stranded.

[0252] siNA sense strand Any of the siNA molecules described herein can comprise a sense strand. The sense strand can comprise a first nucleotide sequence. The first nucleotide sequence can be 15-30, 15-25, 15-23, 17-23, 19-23, or 19-21 nucleotides in length. In some embodiments, the first nucleotide sequence is 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length. In some embodiments, the first nucleotide sequence is at least 19 nucleotides in length. In some embodiments, the first nucleotide sequence is at least 21 nucleotides in length.

[0253] In some embodiments, the sense strand is the same length as the first nucleotide sequence. In some embodiments, the sense strand is longer than the first nucleotide sequence. In some embodiments, the sense strand may further comprise 1, 2, 3, 4, or 5 or more nucleotides than the first nucleotide sequence. In some embodiments, the sense strand may further comprise deoxyribonucleic acid (DNA). In some embodiments, the DNA is thymine (T). In some embodiments, the sense strand may further comprise a TT sequence. In some embodiments, the sense strand may further comprise one or more modified nucleotides adjacent to the first nucleotide sequence. In some embodiments, the one or more modified nucleotides are independently selected from any of the modified nucleotides disclosed herein (e.g., 2'-fluoro nucleotides, 2'-O-methyl nucleotides, 2'-fluoro nucleotide mimics, 2'-O-methyl nucleotide mimics, or nucleotides containing modified nucleobases).

[0254] In some embodiments, the first nucleotide sequence comprises 15, 16, 17, 18, 19, 20, 21, 22, or 23 or more modified nucleotides independently selected from 2'-O-methyl nucleotides and 2'-fluoro nucleotides. In some embodiments, the first nucleotide sequence comprises 16, 17, 18, 19, 20, 21, 22, or 23 or more modified nucleotides independently selected from 2'-O-methyl nucleotides and 2'-fluoro nucleotides. In some embodiments, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides in the first nucleotide sequence are modified nucleotides independently selected from 2'-O-methyl nucleotides and 2'-fluoro nucleotides. In some embodiments, 100% of the nucleotides in the first nucleotide sequence are modified nucleotides independently selected from 2'-O-methyl nucleotides and 2'-fluoro nucleotides. In some embodiments, the 2'-O-methyl nucleotides are 2'-O-methyl nucleotide mimics. In some embodiments, the 2'-fluoro nucleotide is a 2'-fluoro nucleotide mimic.

[0255] In some embodiments, about 15-30, 15-25, 15-24, 15-23, 15-22, 15-21, 17-30, 17-25, 17-24, 17-23, 17-22, 17-21, 18-30, 18-25, 18-24, 18-23, 18-22, 18-21, 19-30, 19-25, 19-24, 19-23, 19-22, 19-21, 20-25, 20-24, 20-23, 21-25, 21-24, or 21-23 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, about 2-20 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, about 5-25 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, about 10-25 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, about 12-25 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least about 12 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least about 13 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least about 14 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least about 15 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least about 16 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least about 17 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least about 18 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides.In some embodiments, at least about 19 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 25, no more than 24, no more than 23, no more than 22, no more than 21, no more than 20, no more than 19, no more than 18, no more than 17, no more than 16, no more than 15, no more than 14, no more than 13, no more than 12, no more than 11, no more than 10, no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, or no more than 2 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 21 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 20 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 19 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 18 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 17 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 16 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 15 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 14 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 13 modified nucleotides of the first nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least one modified nucleotide of the first nucleotide sequence is a 2'-O-methyl pyrimidine. In some embodiments, at least 5, 6, 7, 8, 9, or 10 modified nucleotides of the first nucleotide sequence are 2'-O-methyl pyrimidines. In some embodiments, at least one modified nucleotide of the first nucleotide sequence is a 2'-O-methyl purine. In some embodiments, at least 5, 6, 7, 8, 9, or 10 modified nucleotides of the first nucleotide sequence are 2'-O-methyl purines.In some embodiments, the 2'-O-methyl nucleotide is a 2'-O-methyl nucleotide mimic.

[0256] In some embodiments, 2 to 15 modified nucleotides of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, 2 to 10 modified nucleotides of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, 2 to 6 modified nucleotides of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, 1 to 6, 1 to 5, 1 to 4, or 1 to 3 modified nucleotides of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least 1, 2, 3, 4, 5, or 6 modified nucleotides of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least 2, 3, 4, 5, or 6 modified nucleotides of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least one modified nucleotide of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, at least two modified nucleotides of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least three modified nucleotides of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least four modified nucleotides in the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least five modified nucleotides in the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least six modified nucleotides in the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 10, 9, 8, 7, 6, 5, 4, or 3 modified nucleotides in the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 10, 9, 8, 7, 6, 5, 4, 3, or 2 modified nucleotides in the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 10 modified nucleotides in the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 7 modified nucleotides in the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 6 modified nucleotides in the first nucleotide sequence are 2'-fluoro nucleotides.In some embodiments, no more than 5 modified nucleotides in the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 4 modified nucleotides in the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 3 modified nucleotides in the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 2 modified nucleotides in the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least one modified nucleotide in the first nucleotide sequence is a 2'-fluoro pyrimidine. In some embodiments, 1, 2, 3, 4, 5, or 6 modified nucleotides in the first nucleotide sequence are 2'-fluoro pyrimidines. In some embodiments, at least one modified nucleotide in the first nucleotide sequence is a 2'-fluoro purine. In some embodiments, 1, 2, 3, 4, 5, or 6 modified nucleotides in the first nucleotide sequence are 2'-fluoro purines. In some embodiments, the 2'-fluoro nucleotides are 2'-fluoro nucleotide mimics.

[0257] In some embodiments, the nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least two nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least three nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least four nucleotides from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides: 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19. In some embodiments, at least five nucleotides from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides: 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19. In some embodiments, the nucleotides from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides: 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19. In some embodiments, the nucleotides from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, the nucleotide at position 3 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 7 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 8 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 9 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 12 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 17 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, the 2'-fluoro nucleotide is a 2'-fluoro nucleotide mimic.

[0258] In some embodiments, at least 1, 2, 3, 4, 5, 6, or 7 nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least two nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least three nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, and / or 19 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, the nucleotide at position 3 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 5 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 7 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 8 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 9 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 10 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 11 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 12 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide.In some embodiments, the nucleotide at position 14 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 17 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, the nucleotide at position 19 from the 5' end of the first nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, at least 1, 2, 3, 4, 5, 6, or 7 nucleotides at positions 3, 5, 7, 8, 9, 10, 11, 12, and / or 17 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 3, 7, 8, 9, 12, and / or 17 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 3, 7, 8, 9, 12, and / or 17 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 3, 7, 8, 9, 12, and / or 17 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 5, 7, 8, and / or 9 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 5, 9, 10, 11, 12, and / or 19 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 5, 9, 10, 11, 14, and / or 19 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 5, 9, 10, 11, 14, and / or 19 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, the nucleotides at positions 5, 9, 10, and / or 11 from the 5' end of the first nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, the 2'-fluoro nucleotides are 2'-fluoro nucleotide mimics.

[0259] In some embodiments, the 2'-fluoro nucleotide mimic or the 2'-O-methyl nucleotide mimic has the formula (V): [ka] where R x are independently a nucleobase, aryl, heteroaryl, or H, and Q 1 and Q 2 are independently S or O, and R 5 are independently -OCD3, -F, or -OCH3, and R 6 and R 7 is independently H, D, or CD3. In some embodiments, the nucleobase is selected from thymine, cytosine, guanine, adenine, uracil, and analogs or derivatives thereof.

[0260] In some embodiments, the 2'-fluoro nucleotide mimic or 2'-O-methyl nucleotide mimic has Formula (16)-Formula (20): [ka] where R x are independently a nucleobase, and R 2 is F or —OCH 3 . In some embodiments, the nucleobase is selected from thymine, cytosine, guanine, adenine, uracil, and analogs or derivatives thereof.

[0261] In some embodiments, the sense strand has the following chemical structure: [ka] [where B and R x is a nucleobase, aryl, heteroaryl, or H. In some embodiments, the nucleobases are selected from thymine, cytosine, guanine, adenine, uracil, and analogs or derivatives thereof.

[0262] In some embodiments, the sense strand has the following chemical structure: [ka] [where R y is a nucleobase] In some embodiments, the nucleobases are selected from thymine, cytosine, guanine, adenine, uracil, and analogs or derivatives thereof.

[0263] In some embodiments, the first nucleotide sequence comprises, consists of, or consists essentially of ribonucleic acid (RNA). In some embodiments, the first nucleotide sequence comprises, consists of, or consists essentially of modified RNA. In some embodiments, the modified RNA is selected from 2'-O-methyl RNA and 2'-fluoro RNA. In some embodiments, 15, 16, 17, 18, 19, 20, 21, 22, or 23 modified nucleotides of the first nucleotide sequence are independently selected from 2'-O-methyl RNA and 2'-fluoro RNA.

[0264] In some embodiments, the sense strand may further comprise one or more internucleoside linkages independently selected from phosphodiester (PO) internucleoside linkages, phosphorothioate (PS) internucleoside linkages, mesyl phosphoramidate internucleoside linkages (Ms), phosphorodithioate internucleoside linkages, and PS mimetic internucleoside linkages. In some embodiments, the PS mimetic internucleoside linkage is a sulfonucleoside internucleoside linkage.

[0265] In some embodiments, the sense strand may further comprise at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 or more phosphorothioate internucleoside linkages. In some embodiments, the sense strand comprises no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, or 3 phosphorothioate internucleoside linkages. In some embodiments, the sense strand comprises 2 to 10, 2 to 8, 2 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2 phosphorothioate internucleoside linkages. In some embodiments, the sense strand comprises 1 to 2 phosphorothioate internucleoside linkages. In some embodiments, the sense strand comprises 2 to 4 phosphorothioate internucleoside linkages. In some embodiments, at least one phosphorothioate internucleoside linkage is present between nucleotides 1 and 2 from the 5' end of the first nucleotide sequence. In some embodiments, at least one phosphorothioate internucleoside linkage is present between nucleotides 2 and 3 from the 5' end of the first nucleotide sequence. In some embodiments, the sense strand comprises two phosphorothioate internucleoside linkages between nucleotides 1 through 3 from the 5' end of the first nucleotide sequence.

[0266] In some embodiments, the sense strand may further comprise at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 or more mesyl phosphoramidate internucleoside linkages. In some embodiments, the sense strand comprises no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, or 3 mesyl phosphoramidate internucleoside linkages. In some embodiments, the sense strand comprises 2 to 10, 2 to 8, 2 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2 mesyl phosphoramidate internucleoside linkages. In some embodiments, the sense strand comprises 1 to 2 mesyl phosphoramidate internucleoside linkages. In some embodiments, the sense strand comprises 2-4 mesyl phosphoramidate internucleoside linkages.

[0267] In some embodiments, the sense strand can include any of the modified nucleotides disclosed in the subsection below entitled "Modified Nucleotides." In some embodiments, the sense strand can include a 5'-stabilized endcap, which can be selected from those disclosed in the subsection below entitled "5'-stabilized endcaps."

[0268] In some embodiments, any of the sense strands disclosed herein further comprises a TT sequence adjacent to the first nucleotide sequence.

[0269] siNA antisense strand Any of the siNA molecules described herein can comprise an antisense strand. The antisense strand can comprise a second nucleotide sequence. The second nucleotide sequence can be 15-30, 15-25, 15-23, 17-23, 19-23, or 19-21 nucleotides in length. In some embodiments, the second nucleotide sequence is 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length. In some embodiments, the second nucleotide sequence is at least 19 nucleotides in length. In some embodiments, the second nucleotide sequence is at least 21 nucleotides in length.

[0270] In some embodiments, the antisense strand is the same length as the second nucleotide sequence. In some embodiments, the antisense strand is longer than the second nucleotide sequence. In some embodiments, the antisense strand may further comprise 1, 2, 3, 4, or 5 or more nucleotides than the second nucleotide sequence. In some embodiments, the antisense strand is the same length as the sense strand. In some embodiments, the antisense strand is longer than the sense strand. In some embodiments, the antisense strand may further comprise 1, 2, 3, 4, or 5 or more nucleotides than the sense strand. In some embodiments, the antisense strand may further comprise deoxyribonucleic acid (DNA). In some embodiments, the DNA is thymine (T). In some embodiments, the antisense strand may further comprise a TT sequence. In some embodiments, the antisense strand may further comprise one or more modified nucleotides adjacent to the second nucleotide sequence. In some embodiments, the one or more modified nucleotides are independently selected from any of the modified nucleotides disclosed herein (e.g., a 2'-fluoro nucleotide, a 2'-O-methyl nucleotide, a 2'-fluoro nucleotide mimic, a 2'-O-methyl nucleotide mimic, or a nucleotide comprising a modified nucleobase).

[0271] In some embodiments, the second nucleotide sequence comprises 15, 16, 17, 18, 19, 20, 21, 22, or 23 or more modified nucleotides independently selected from 2'-O-methyl nucleotides and 2'-fluoro nucleotides. In some embodiments, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides in the second nucleotide sequence are modified nucleotides independently selected from 2'-O-methyl nucleotides and 2'-fluoro nucleotides. In some embodiments, 100% of the nucleotides in the second nucleotide sequence are modified nucleotides independently selected from 2'-O-methyl nucleotides and 2'-fluoro nucleotides.

[0272] In some embodiments, about 15-30, 15-25, 15-24, 15-23, 15-22, 15-21, 17-30, 17-25, 17-24, 17-23, 17-22, 17-21, 18-30, 18-25, 18-24, 18-23, 18-22, 18-21, 19-30, 19-25, 19-24, 19-23, 19-22, 19-21, 20-25, 20-24, 20-23, 21-25, 21-24, or 21-23 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, about 2-20 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, about 5-25 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, about 10-25 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, about 12-25 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, the second nucleotide sequence comprises 16, 17, 18, 19, 20, 21, 22, or 23 or more modified nucleotides independently selected from 2'-O-methyl nucleotides and 2'-fluoro nucleotides. In some embodiments, at least about 12 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least about 13 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least about 14 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least about 15 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least about 16 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides.In some embodiments, at least about 17 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least about 18 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least about 19 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 25, no more than 24, no more than 23, no more than 22, no more than 21, no more than 20, no more than 19, no more than 18, no more than 17, no more than 16, no more than 15, no more than 14, no more than 13, no more than 12, no more than 11, no more than 10, no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, or no more than 2 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 21 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 20 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 19 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 18 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 17 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 16 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 15 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 14 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, no more than 13 modified nucleotides of the second nucleotide sequence are 2'-O-methyl nucleotides. In some embodiments, at least one modified nucleotide of the second nucleotide sequence is a 2'-O-methyl pyrimidine. In some embodiments, at least 5, 6, 7, 8, 9, or 10 modified nucleotides of the second nucleotide sequence are 2'-O-methyl pyrimidines.In some embodiments, at least one modified nucleotide of the second nucleotide sequence is a 2'-O-methyl purine. In some embodiments, at least 5, 6, 7, 8, 9, or 10 modified nucleotides of the second nucleotide sequence are 2'-O-methyl purines. In some embodiments, the 2'-O-methyl nucleotides are 2'-O-methyl nucleotide mimics.

[0273] In some embodiments, 2 to 15 modified nucleotides of the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, 2 to 10 modified nucleotides of the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, 2 to 6 modified nucleotides of the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, 1 to 6, 1 to 5, 1 to 4, or 1 to 3 modified nucleotides of the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least 1, 2, 3, 4, 5, or 6 modified nucleotides of the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least 2, 3, 4, 5, or 6 modified nucleotides of the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least 1 modified nucleotide of the second nucleotide sequence is a 2'-fluoro nucleotide. In some embodiments, at least 2 modified nucleotides of the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least 3 modified nucleotides of the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least four modified nucleotides in the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least five modified nucleotides in the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 10, 9, 8, 7, 6, 5, 4, or 3 modified nucleotides in the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 10, 9, 8, 7, 6, 5, 4, 3, or 2 modified nucleotides in the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 10 modified nucleotides in the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 7 modified nucleotides in the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 6 modified nucleotides in the second nucleotide sequence are 2'-fluoro nucleotides.In some embodiments, no more than 5 modified nucleotides of the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 4 modified nucleotides of the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 3 modified nucleotides of the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, no more than 2 modified nucleotides of the second nucleotide sequence are 2'-fluoro nucleotides. In some embodiments, at least one modified nucleotide of the second nucleotide sequence is a 2'-fluoro pyrimidine. In some embodiments, 1, 2, 3, 4, 5, or 6 modified nucleotides of the second nucleotide sequence are 2'-fluoro pyrimidines. In some embodiments, at least one modified nucleotide of the second nucleotide sequence is a 2'-fluoro purine. In some embodiments, 1, 2, 3, 4, 5, or 6 modified nucleotides of the second nucleotide sequence are 2'-fluoro purines. In some embodiments, the 2'-fluoro nucleotides are 2'-fluoro nucleotide mimics.

[0274] In some embodiments, the 2'-fluoro nucleotide or 2'-O-methyl nucleotide is a 2'-fluoro nucleotide mimic or a 2'-O-methyl nucleotide mimic. In some embodiments, the 2'-fluoro nucleotide mimic or the 2'-O-methyl nucleotide mimic has the formula (V): [ka] where R x are independently a nucleobase, aryl, heteroaryl, or H, and Q 1 and Q 2 are independently S or O, and R 5 are independently -OCD3, -F, or -OCH3, and R 6 and R 7 is independently H, D, or CD3. In some embodiments, the nucleobase is selected from thymine, cytosine, guanine, adenine, uracil, and analogs or derivatives thereof.

[0275] In some embodiments, the 2'-fluoro nucleotide mimic or 2'-O-methyl nucleotide mimic has Formula (16)-Formula (20): [ka] where R x is a nucleobase, and R 2 is independently F or —OCH 3 . In some embodiments, the nucleobase is selected from thymine, cytosine, guanine, adenine, uracil, and analogs or derivatives thereof.

[0276] In some embodiments, the antisense strand has the following chemical structure: [ka] [where B and R x is a nucleobase, aryl, heteroaryl, or H. In some embodiments, the nucleobases are selected from thymine, cytosine, guanine, adenine, uracil, and analogs or derivatives thereof.

[0277] In some embodiments, the antisense strand has the following chemical structure: [ka] [where R y is a nucleobase] In some embodiments, the nucleobases are selected from thymine, cytosine, guanine, adenine, uracil, and analogs or derivatives thereof.

[0278] In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, or 9 nucleotides at positions 2, 5, 6, 8, 10, 14, 16, 17, and / or 18 from the 5' end of the second nucleotide sequence are 2'-fluoro nucleo...

Claims

1. A double-stranded short interfering nucleic acid (siNA) molecule comprising: (a) a sense strand comprising a nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% identical to the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603 or 638; and / or (b) an antisense strand comprising a nucleotide sequence that is at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644; Including, wherein the siNA molecule down-regulates the expression of the hydroxysteroid 17-beta dehydrogenase 13 (HSD17B13) gene.

2. A double-stranded short interfering nucleic acid (siNA) molecule comprising: (a) a sense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 1-100, 201-230, 262-287, 314-445, 576-603, or 638; and / or (b) an antisense strand comprising the nucleotide sequence of any one of SEQ ID NOs: 101-200, 231-260, 288-313, 446-575, 604-637, or 639-644; Including, wherein the siNA molecule down-regulates the expression of the hydroxysteroid 17-beta dehydrogenase 13 (HSD17B13) gene.

3. The siNA molecule of claim 2, wherein the sense strand and / or the antisense strand comprises at least one modified nucleotide selected from the group consisting of 2'-O-methyl, 2'-fluoro, locked nucleic acid, nucleoside analog, 5'-terminal vinyl phosphonate, and 5'-phosphorothioate internucleoside linkage.

4. The siNA molecule of claim 2, wherein the siNA molecule is selected from any one of siNA duplex SEQ ID NOs: D1-D178 or MD1-MD178.

5. A pharmaceutical composition comprising the siNA molecule of any one of claims 1 to 4.

6. 6. The pharmaceutical composition of claim 5, further comprising at least one additional active agent, wherein the at least one additional active agent is a liver disease therapeutic agent.

7. 7. The pharmaceutical composition of claim 6, wherein the liver disease therapeutic agent is selected from peroxisome proliferator-activated receptor (PPAR) agonists, farnesoid X receptor (FXR) agonists, lipid-altering agents, incretin-based therapies, and thyroid hormone receptor (THR) modulators.

8. Use of a siNA molecule of any one of claims 1 to 4 in the preparation of a medicament for the treatment of liver disease in a subject in need thereof.

9. 10. Use of the pharmaceutical composition of claim 5 in the preparation of a medicament for the treatment of liver disease in a subject in need thereof.

10. 9. The use according to claim 8, wherein the liver disease is non-alcoholic fatty liver disease (NAFLD).

11. 9. The use according to claim 8, wherein the liver disease is non-alcoholic steatohepatitis (NASH).

12. 9. The use of claim 8, wherein the medicament further comprises at least one additional active agent, wherein the at least one additional active agent is a liver disease therapeutic agent.

13. Use of a siNA molecule of any one of claims 1 to 4 in the preparation of a medicament for reducing the expression level of HSD17B13 in a subject in need thereof.

14. Use of a siNA molecule of any one of claims 1 to 4 in the preparation of a medicament for preventing at least one symptom of liver disease in a subject in need thereof.

15. The siNA molecule of any one of claims 1 to 4, further comprising a ligand.

16. The ligand is 【Chemical 1】 The siNA molecule of claim 15, wherein:

17. The siNA molecule of claim 2, wherein the sense strand is selected from any of SEQ ID NOs: 316-445, 576-603, or 638.

18. The siNA molecule of claim 2, wherein the antisense strand is selected from any of SEQ ID NOs: 446-575, 604-637, or 639-644.

19. 10. Use of the pharmaceutical composition of claim 5 in the preparation of a medicament for reducing the expression level of HSD17B13 in a subject in need thereof.

20. 10. Use of the pharmaceutical composition of claim 5 in the preparation of a medicament for preventing at least one symptom of liver disease in a subject in need thereof.

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