HSD17B13-Related Double-Stranded Oligonucleotide Compositions and Related Methods

Abstract

The present disclosure provides HSD17B13-related double-stranded oligonucleotides, compositions, and methods of using such double-stranded oligonucleotides and compositions to prevent and / or treat various conditions, disorders, or diseases associated with expression of HSD17B13. In some embodiments, the double-stranded oligonucleotides and compositions provided include nucleobase modifications, sugar modifications, internucleotide linkage modifications and / or patterns thereof to improve specificity, activity and / or selectivity. In some embodiments, the double-stranded oligonucleotides and compositions provided target HSD17B13.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 264,360, filed November 19, 2021, U.S. Provisional Application No. 63 / 268,775, filed March 2, 2022, and U.S. Provisional Application No. 63 / 377,482, filed September 28, 2022, the contents of each of which are incorporated by reference in their entirety and priority is claimed.

[0002] Technical Field The present disclosure provides, inter alia, double-stranded (ds) oligonucleotides, compositions, and methods (e.g., preparation, use, etc.) thereof. In some embodiments, the provided technology is useful for the prevention and / or treatment of various conditions, disorders, or diseases associated with expression of 17-beta hydroxysteroid dehydrogenase 13 (HSD17B13). [Background technology]

[0003] background Double-stranded (ds) oligonucleotides are useful in a variety of applications, including therapeutic, diagnostic, and / or research applications. For example, ds oligonucleotides targeting HSD17B13 may be useful in treating conditions, disorders, or diseases associated with HSD17B13 expression, such as non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), alcoholic steatohepatitis (ASH), alcoholic liver disease, non-alcoholic liver disease, alcoholic cirrhosis, non-alcoholic cirrhosis, steatohepatitis, hepatic steatosis, hepatocellular carcinoma, HCV hepatitis, chronic hepatitis, hereditary hemochromatosis, primary sclerosing cholangitis, drug-induced liver injury, or hepatocellular necrosis. Summary of the Invention [Means for solving the problem]

[0004] overview In some embodiments, the present disclosure provides ds oligonucleotides and compositions thereof targeting HSD17B13 with significantly improved properties and / or enhanced activity. Among other things, the present disclosure provides techniques for designing, producing, and utilizing such ds oligonucleotides and compositions. In particular, in some embodiments, the present disclosure provides ds oligonucleotides comprising useful internucleotide linkage patterns and / or sugar modification patterns that, when combined with one or more other structural elements, such as base sequence (or a portion thereof), nucleobase modifications (and patterns thereof), additional chemical moieties, etc., can provide ds oligonucleotides and compositions thereof targeting HSD17B13 with enhanced activity and / or desirable properties, including, but not limited to, effective and efficient reduction of the expression, level, and / or activity of HSD17B13 transcripts and products encoded thereby. In some embodiments, ds oligonucleotides and compositions targeting HSD17B13 reduce the level of HSD17B13 transcripts and are useful for treating and / or preventing HSD17B13-related conditions, disorders or diseases, such as NAFLD, NASH, ASH, alcoholic liver disease, non-alcoholic liver disease, alcoholic cirrhosis, non-alcoholic cirrhosis, steatohepatitis, hepatic steatosis, hepatocellular carcinoma, HCV hepatitis, chronic hepatitis, hereditary hemochromatosis, primary sclerosing cholangitis, drug-induced liver injury or hepatocellular necrosis.

[0005] In some embodiments, ds oligonucleotides targeting HSD17B13 are capable of mediating knockdown of HSD17B13 such that the level, expression, and / or activity of HSD17B13 or its products is reduced. In some embodiments, ds oligonucleotides targeting HSD17B13 are capable of mediating pan-specific knockdown of HSD17B13 such that the level, expression, and / or activity of multiple or all HSD17B13 alleles is reduced. In some embodiments, ds oligonucleotides targeting HSD17B13 have a base sequence complementary to a sequence common to multiple or all HSD17B13 alleles.

[0006] In certain embodiments, such structural elements include one or more of: (1) chemical modifications (e.g., modifications of sugars, bases, and / or internucleotide linkages) and their patterns; and (2) changes in stereochemistry (e.g., the stereochemistry of backbone chiral internucleotide linkages) and their patterns. In certain embodiments, one or more of such structural elements can be independently present in one or both oligonucleotides of a ds oligonucleotide. In certain embodiments, properties and / or activities influenced by such structural elements include, but are not limited to, involvement in, or directionality of, a reduction in the expression, activity, or level of a gene or its gene product, for example, mediated by RNA interference (RNAi interference).

[0007] In certain embodiments, the present disclosure demonstrates that compositions comprising ds oligonucleotides (e.g., dsRNAi oligonucleotides (also called dsRNAi agents)) with controlled structural elements provide unexpected properties and / or activities.

[0008] In certain embodiments, the present disclosure encompasses the recognition that stereochemistry, e.g., the stereochemistry of backbone chiral centers, can unexpectedly maintain or improve the properties of ds oligonucleotides. For example, but not by way of limitation, the present disclosure provides, in part, (1) a guide strand containing a backbone phosphorothioate chiral center in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the (N-2) nucleotide immediately upstream, i.e., in the 5' direction; (2) a guide strand containing Rp, Sp, or alternating backbone phosphorothioate chiral centers between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide in the 3' direction, and between the +2 nucleotide and the immediately downstream (+3) nucleotide; (3) a guide strand that contains one or more backbone phosphorothioate chiral centers in the Sp configuration upstream, i.e., in the 5' direction, of a backbone phosphorothioate chiral center between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide, wherein the upstream backbone phosphorothioate chiral center is in the Rp or Sp configuration; (4) a guide strand containing one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide, and between (a) the +3 and +4 nucleotides and (b) the +5 and +6 nucleotides; (5) a passenger strand in combination with one or more of the aforementioned guide strands, comprising one or more backbone chiral centers of Rp or Sp configuration; and (6) a passenger strand, combined with one or more of the aforementioned guide strands, containing a backbone phosphorothioate chiral center of the Sp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide in the 3' direction, and between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide; and ds oligonucleotides are (1) A guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, i.e., the guide strand contains one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3'(N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) A guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond exists between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The present invention relates to a ds oligonucleotide, further comprising a 2' modification, such as a 2'F modification, of the 3' nucleotide of a nucleotide pair linked by an Rp, Sp, or stereorandom non-negatively charged internucleotide bond. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide bonds. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds are Sp non-negatively charged internucleotide bonds. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds are stereorandom non-negatively charged internucleotide bonds.

[0009] In certain embodiments, the present disclosure encompasses the recognition that stereochemistry, e.g., the stereochemistry of the chiral center in the 5'-terminal modification of the guide strand, can unexpectedly maintain or improve the properties of the ds oligonucleotides described herein. For example, but not by way of limitation, the present disclosure encompasses, in part, (1) phosphorothioate chiral centers in the Rp or Sp configuration; (2) Rp, Sp, or stereorandom non-negatively charged internucleotide linkages in which the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide linkage contains a 2' modification, e.g., 2'F; and (3) (a) Including, but not limited to: [ka] 5'PO modifications such as; (b) Without limitation, [ka] 5'VP modifications such as; (c) Without limitation, [ka] 5'MeP modifications such as; (d) Without limitation, [ka] 5'PN and 5'triazole-P modifications such as wherein the bases are selected from A, C, G, T, U, abasic and modified nucleobases; R 2’ is H, OH, O-alkyl, F, MOE, locked nucleic acid (LNA) bridges, and bridged nucleic acid (BNA) bridges to the 4'C, including, but not limited to: [ka] (selected from The present invention relates to a ds polynucleotide comprising a guide strand comprising a 5'-end modification selected from:

[0010] In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0011] In certain other embodiments, the present disclosure encompasses the recognition that the stereochemistry, e.g., the stereochemistry of the chiral center at the 5'-terminal nucleotide of the guide strand, can unexpectedly maintain or improve the properties of ds oligonucleotides in which the guide strand of the ds oligonucleotide also contains a phosphorothioate chiral center in the Rp or Sp configuration. For example, but not by way of limitation, the present disclosure encompasses, in part, (1) a phosphorothioate chiral center in the Rp or Sp configuration; (2) an Rp, Sp, or stereorandom non-negatively charged internucleotide linkage in which the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide linkage contains a 2' modification, e.g., 2'F; and (3) (a) Including, but not limited to: [ka] 5'PO nucleotides such as; (b) Without limitation, [ka] 5'VP nucleotides such as; (c) Without limitation, [ka] 5'MeP nucleotides such as; (d) Without limitation, [ka] 5'PN and 5'triazole-P nucleotides such as; (e) Without limitation, [ka] 5' abasic VP and 5' abasic MeP nucleotides such as The present invention relates to a ds polynucleotide comprising a guide strand comprising a 5'-end modification selected from:

[0012] In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0013] In certain embodiments, the present disclosure encompasses the recognition that non-natural internucleotide linkages, such as neutral internucleotide linkages, can be used to attach one or more molecules to the double-stranded oligonucleotides described herein. In certain embodiments, such binding molecules can facilitate targeting and / or delivery of the double-stranded oligonucleotides. For example, but not limited to, such binding molecules include lipophilic molecules. In certain embodiments, the binding molecule is a molecule comprising one or more GalNAc moieties. In certain embodiments, the binding molecule is a receptor. In certain embodiments, the binding molecule is a receptor ligand.

[0014] In certain embodiments, the present disclosure provides techniques for incorporating various additional chemical moieties into ds oligonucleotides. In certain embodiments, the present disclosure provides, for example, reagents and methods for introducing additional chemical moieties with a nucleobase (e.g., by covalent attachment to a site on the nucleobase, optionally via a linker).

[0015] In certain embodiments, the present disclosure provides techniques, e.g., ds oligonucleotide compositions and methods, for achieving allele-specific suppression, in which transcripts from one allele of a particular target gene are selectively knocked down relative to at least one other allele of the same gene.

[0016] In particular, the present disclosure provides structural elements, techniques, and / or features that can be incorporated into ds oligonucleotides and that can confer or adjust one or more properties thereof (e.g., compared to an otherwise identical ds oligonucleotide lacking the related technique or feature). In certain embodiments, the present disclosure describes that one or more of the provided techniques and / or features can be usefully incorporated into ds oligonucleotides of various sequences.

[0017] In certain embodiments, the present disclosure demonstrates that certain provided structural elements, techniques, and / or features are particularly useful for ds oligonucleotides (e.g., RNAi agents) that participate in and / or induce the RNAi mechanism. Nevertheless, however, the teachings of the present disclosure are not limited to ds oligonucleotides that participate in or act by any particular mechanism. In certain embodiments, the present disclosure relates to any ds oligonucleotides that are useful for any purpose, that act via any mechanism, and that include any sequence, structure, or format (or portion thereof) described herein. In certain embodiments, the present disclosure relates to ds oligonucleotides that are useful for any purpose, that act via any mechanism, and that include any sequence, structure, or format (or portion thereof) described herein, (1) a guide strand containing a backbone phosphorothioate chiral center in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the (N-2) nucleotide immediately upstream, i.e., in the 5' direction; (2) a guide strand containing Rp, Sp, or alternating backbone phosphorothioate chiral centers between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide in the 3' direction, and between the +2 nucleotide and the immediately downstream (+3) nucleotide; (3) a guide strand that contains one or more backbone phosphorothioate chiral centers in the Sp configuration upstream, i.e., in the 5' direction, of a backbone phosphorothioate chiral center between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide, wherein the upstream backbone phosphorothioate chiral center is in the Rp or Sp configuration; (4) a guide strand containing one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide, and between (a) the +3 and +4 nucleotides and (b) the +5 and +6 nucleotides; (5) a passenger strand in combination with one or more of the aforementioned guide strands, comprising one or more backbone chiral centers of Rp or Sp configuration; and (6) a passenger strand, combined with one or more of the aforementioned guide strands, containing a backbone phosphorothioate chiral center of the Sp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide in the 3' direction, and between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide; ds oligonucleotide comprising and ds oligonucleotides are (1) A guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand contains one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The present invention relates to a ds oligonucleotide, further comprising a 2' modification, such as a 2'F modification, of the 3' nucleotide of a nucleotide pair linked by an Rp, Sp, or stereorandom non-negatively charged internucleotide bond. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide bonds. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds are Sp non-negatively charged internucleotide bonds. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds are stereorandom non-negatively charged internucleotide bonds.

[0018] In certain embodiments, the provided ds oligonucleotides can participate (eg, directly) in the RNAi machinery.

[0019] In certain embodiments, the guide strand comprises backbone phosphorothioate chiral centers in the Sp configuration between the 3' terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide; (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by an Rp, Sp, or stereorandom non-negatively charged internucleotide bond, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide bonds, where n is about 1 to 49;

[0020] In certain embodiments, the present disclosure demonstrates that compositions comprising ds oligonucleotides (e.g., dsRNAi oligonucleotides (also called dsRNAi agents)) with controlled structural elements provide unexpected properties and / or activities.

[0021] In certain embodiments, the guide strand contains Rp, Sp, or alternating backbone phosphorothioate chiral centers between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide; (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide linkage, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, where n is about 1 to 49. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0022] In certain embodiments, the guide strand comprises one or more backbone phosphorothioate chiral centers of Rp or Sp configuration upstream of a backbone phosphorothioate chiral center of Sp configuration between the 3' terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide; (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide linkage, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, where n is about 1 to 49. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0023] In certain embodiments, the guide strand comprises one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds between the second (+2) nucleotide and the third (+3) nucleotide from the 5'-terminal nucleotide of the guide strand, and an internucleotide bond to the 3'(N-1) nucleotide immediately preceding the terminal end; (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide linkage, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, where n is about 1 to 49. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0024] In certain embodiments, the guide strand comprises backbone phosphorothioate chiral centers in the Sp configuration between the 3' terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide; (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide bond, and the passenger strand comprises one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0025] In certain embodiments, the guide strand contains Rp, Sp, or alternating backbone phosphorothioate chiral centers between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide; (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by an Rp, Sp, or stereorandom non-negatively charged internucleotide bond, and the passenger strand comprises one or more backbone chiral centers in an Rp or Sp configuration. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0026] In certain embodiments, the guide strand comprises one or more backbone phosphorothioate chiral centers of Rp or Sp configuration upstream of the backbone chiral center of Sp configuration between the 3' terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide; (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by an Rp, Sp, or stereorandom non-negatively charged internucleotide bond, and the passenger strand comprises one or more backbone centers in an Rp or Sp configuration. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0027] In certain embodiments, the guide strand contains one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the (+2) nucleotide and the immediately downstream (+3) nucleotide, and between (a) the (+3) nucleotide and the (+4) nucleotide and (b) the (+5) nucleotide and the (+6) nucleotide; (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by an Rp, Sp, or stereorandom non-negatively charged internucleotide bond, and the passenger strand comprises one or more backbone chiral centers in an Rp or Sp configuration. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0028] In certain embodiments, the guide strand comprises one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, or 2' modifications, e.g., 2'F modifications, of the 3' nucleotides of nucleotide pairs linked by Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, present between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3'(N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide), and the passenger strand comprises one or more backbone chiral centers in the Rp or Sp configuration. In certain embodiments, the one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, the one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more of Rp, Sp or stereorandom non-negatively charged internucleotide linkages is a stereorandom non-negatively charged internucleotide linkage.

[0029] In certain embodiments, the guide strand comprises backbone phosphorothioate chiral centers in the Sp configuration between the 3'-terminal nucleotide and the (N-1) nucleotide immediately preceding the terminal end and between the (N-1) nucleotide immediately preceding the terminal end and the (N-2) nucleotide immediately upstream thereof, and 2' modifications, e.g., 2'F modifications, of the 3' nucleotides of nucleotide pairs linked by Rp, Sp, or stereorandom non-negatively charged internucleotide linkages; and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages (where n is about 1 to 49) and one or more backbone chiral centers in the Rp or Sp configuration. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more of Rp, Sp or stereorandom non-negatively charged internucleotide linkages is a stereorandom non-negatively charged internucleotide linkage.

[0030] In certain embodiments, the guide strand comprises Rp, Sp, or alternating backbone phosphorothioate chiral centers between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the (+2) nucleotide and the immediately downstream (+3) nucleotide, and 2' modifications, e.g., 2'F modifications, of the 3' nucleotides of nucleotide pairs linked by Rp, Sp, or stereorandom non-negatively charged internucleotide linkages; and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages (where n is about 1 to 49) and one or more backbone chiral centers in the Rp or Sp configuration. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more of Rp, Sp or stereorandom non-negatively charged internucleotide linkages is a stereorandom non-negatively charged internucleotide linkage.

[0031] In certain embodiments, the guide strand comprises one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration upstream of the backbone phosphorothioate chiral center in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide, and a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide linkages; and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages (where n is about 1 to 49) and one or more backbone chiral centers in the Rp or Sp configuration. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more of Rp, Sp or stereorandom non-negatively charged internucleotide linkages is a stereorandom non-negatively charged internucleotide linkage.

[0032] In certain embodiments, the guide strand comprises one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, or 2' modifications, e.g., 2'F modifications, of the 3' nucleotide of a nucleotide pair linked by an Rp, Sp, or stereorandom non-negatively charged internucleotide linkage, present between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3'(N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide), and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages (where n is about 1 to 49) and one or more backbone chiral centers of an Rp or Sp configuration. In certain embodiments, the one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, the one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more of Rp, Sp or stereorandom non-negatively charged internucleotide linkages is a stereorandom non-negatively charged internucleotide linkage.

[0033] In certain embodiments, provided ds oligonucleotides may be involved in exon skipping mechanisms. In certain embodiments, provided ds oligonucleotides may be aptamers. In certain embodiments, provided ds oligonucleotides may bind to and inhibit the function of proteins, small molecules, nucleic acids, or cells. In certain embodiments, provided ds oligonucleotides may be involved in the formation of triple helices with double-stranded nucleic acids within cells. In certain embodiments, provided ds oligonucleotides may bind to genomic (e.g., chromosomal) nucleic acids. In certain embodiments, provided ds oligonucleotides may bind to genomic (e.g., chromosomal) nucleic acids and thus prevent or reduce expression of the nucleic acid (e.g., by preventing or reducing transcription, transcription promotion, modification, etc.). In certain embodiments, provided ds oligonucleotides may bind to DNA quadruplexes. In certain embodiments, provided ds oligonucleotides may be immunomodulatory. In certain embodiments, provided ds oligonucleotides may be immunostimulatory. In certain embodiments, provided oligonucleotides may be immunostimulatory and may include CpG sequences. In certain embodiments, provided ds oligonucleotides may be immunostimulatory, may contain CpG sequences, and may be useful as adjuvants. In certain embodiments, provided ds oligonucleotides may be immunostimulatory, may contain CpG sequences, and may be useful as adjuvants in the treatment of disease (e.g., infectious disease or cancer). In certain embodiments, provided ds oligonucleotides may be therapeutic. In certain embodiments, provided ds oligonucleotides may be non-therapeutic. In certain embodiments, provided ds oligonucleotides may be therapeutic or non-therapeutic. In certain embodiments, provided ds oligonucleotides may be useful for therapeutic, diagnostic, research, and / or nanomaterial applications. In certain embodiments, provided ds oligonucleotides may be useful for experimental purposes. In certain embodiments, provided ds oligonucleotides may be useful for experimental purposes, for example, as probes, in microarrays, etc.In certain embodiments, provided ds oligonucleotides may participate in more than one biological mechanism; in certain such embodiments, for example, provided ds oligonucleotides may participate in both the RNAi and RNase H mechanisms.

[0034] In certain embodiments, the provided ds oligonucleotides are directed to an HSD17B13 target (e.g., an HSD17B13 target sequence, an HSD17B13 target RNA, an HSD17B13 target mRNA, an HSD17B13 target pre-mRNA, an HSD17B13 target gene, etc.). An HSD17B13 target gene is a gene for which the expression and / or activity of one or more HSD17B13 gene products (e.g., HSD17B13 RNA and / or protein products) is intended to be altered. In certain embodiments, the HSD17B13 target gene is intended to be inhibited. Thus, when the ds oligonucleotides described herein act on an HSD17B13 target gene, the presence and / or activity of one or more HSD17B13 gene products is altered in the presence of the ds oligonucleotide compared to when the ds oligonucleotide is absent.

[0035] In certain embodiments, an HSD17B13 target is a specific HSD17B13 allele for which the expression and / or activity of one or more products (e.g., HSD17B13 RNA and / or protein products) is intended to be altered. In certain embodiments, the HSD17B13 target allele is one whose presence and / or expression is associated with (e.g., correlates with) the presence, incidence, and / or severity of one or more HSD17B13-associated diseases and / or conditions. Alternatively, or in addition, in certain embodiments, the HSD17B13 target allele is one for which alteration of the level and / or activity of one or more HSD17B13 gene products is correlated with amelioration of one or more aspects of an HSD17B13-associated disease and / or condition (e.g., delayed onset, reduced severity, responsiveness to other therapies, etc.).

[0036] In certain embodiments, for example, when the presence and / or activity of a particular HSD17B13 allele (HSD17B13 disease-associated allele) is associated (e.g., correlated) with the presence, occurrence and / or severity of one or more disorders, diseases and / or conditions, and a different HSD17B13 allele exists that is unassociated or less associated (e.g., exhibits a less significant or statistically insignificant correlation), the ds oligonucleotides and methods described herein may preferentially or specifically target the associated allele relative to one or more less associated / unassociated alleles, and thus mediate allele-specific suppression.

[0037] In certain embodiments, the HSD17B13 target sequence is an HSD17B13 sequence to which an oligonucleotide described herein binds. In certain embodiments, the HSD17B13 target sequence is identical to or an exact complement of the HSD17B13 sequence of a provided oligonucleotide or consecutive residues therein (e.g., a provided oligonucleotide comprises an HSD17B13 target binding sequence that is identical to or an exact complement of the HSD17B13 target sequence). In certain embodiments, the HSD17B13 target binding sequence is an exact complement of the HSD17B13 target sequence of an HSD17B13 transcript (e.g., pre-mRNA, mRNA, etc.). The HSD17B13 target binding sequence / target sequence can be of various lengths for a provided oligonucleotide with desired activity and / or properties. In certain embodiments, the HSD17B13 target binding sequence / target sequence comprises 5-50 (e.g., 10-40, 15-30, 15-25, 16-25, 17-25, 18-25, 19-25, 20-25, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more) bases. In certain embodiments, a small number of differences / mismatches are tolerated between (relevant portions of) the oligonucleotide and its target sequence, including but not limited to the HSD17B13 target and / or 5' and / or 3' terminal regions of the oligonucleotide sequence. In certain embodiments, the HSD17B13 target sequence is present within an HSD17B13 target gene. In certain embodiments, the HSD17B13 target sequence is present within an HSD17B13 transcript (eg, mRNA and / or pre-mRNA) produced from an HSD17B13 target gene.

[0038] In certain embodiments, the HSD17B13 target sequence comprises one or more allelic sites (i.e., positions at which allelic variations occur within the HSD17B13 target gene). In certain embodiments, the allelic sites are variants. In certain embodiments, the allelic sites are SNPs. In some such embodiments, provided oligonucleotides bind preferentially or specifically to one allele over one or more other alleles. In certain embodiments, provided oligonucleotides bind preferentially to disease-associated alleles. For example, in certain embodiments, the oligonucleotides provided herein (or target binding sequence portions thereof) have a sequence that is completely or at least partially identical to or an exact complement of a particular allelic version of an HSD17B13 target sequence.

[0039] In certain embodiments, the oligonucleotides provided herein (or their target binding sequence portions) have a sequence that is identical to or the exact complement of an HSD17B13 target sequence comprising an allele or an allelic site of a disease-associated allele. In certain embodiments, the oligonucleotides provided herein have an HSD17B13 target binding sequence that is the exact complement of an HSD17B13 target sequence comprising an allelic site of an HSD17B13 transcript of an allele (in certain embodiments, a disease-associated allele), where the allelic site is a variant. In certain embodiments, the oligonucleotides provided herein have an HSD17B13 target binding sequence that is the exact complement of an HSD17B13 target sequence comprising an allelic site of an HSD17B13 transcript of an allele (in certain embodiments, a disease-associated allele), where the allelic site is a SNP. In certain embodiments, the sequence is any sequence disclosed herein.

[0040] Unless otherwise indicated, all sequences (including, but not limited to, base sequences and chemical, modification and / or stereochemical patterns) are presented in 5' to 3' order, with the 5' terminal nucleotide identified as the "+1" position, the 3' terminal nucleotide identified by the number of the nucleotide in the complete sequence or "N", the pre-terminal nucleotide identified as, for example, "N-1", etc.

[0041] In certain embodiments, the present disclosure provides compositions and methods relating to oligonucleotides that are specific for HSD17B13 targets and have any of the formats, structural elements, or base sequences of any of the oligonucleotides disclosed herein.

[0042] In certain embodiments, the present disclosure provides compositions and methods relating to oligonucleotides that are specific for an HSD17B13 target and have or include the base sequence of any oligonucleotide disclosed herein or a region of at least 15 contiguous nucleotides of the base sequence of any oligonucleotide disclosed herein, wherein the first nucleotide of the base sequence or the first nucleotide of the at least 15 contiguous nucleotides can optionally be replaced by a T or a DNA T.

[0043] In certain embodiments, the present disclosure provides compositions and methods for RNA interference induced by RNAi agents (also called RNAi oligonucleotides). In certain embodiments, the oligonucleotides of such compositions can have the format, structural elements, or base sequence of the oligonucleotides disclosed herein.

[0044] In certain embodiments, the present disclosure provides compositions and methods for RNase H-mediated knockdown of HSD17B13 target gene RNA induced by oligonucleotides (eg, antisense oligonucleotides).

[0045] The provided oligonucleotides and oligonucleotide compositions can have any format, structural element, or base sequence of any of the oligonucleotides disclosed herein. In certain embodiments, the structural element is a 5'-terminal structure, a 5'-terminal region, a 5'-nucleotide, a seed region, a post-seed region, a 3'-terminal region, a 3'-terminal dinucleotide, a 3'-end cap, or any portion of these structures, GC content, a long GC stretch, and / or any modification, chemistry, stereochemistry, pattern of modification, chemistry or stereochemistry, or chemical moiety (e.g., including, but not limited to, a targeting moiety, a lipid moiety, a GalNAc moiety, a carbohydrate moiety, etc.), any component, or any combination of any of the above.

[0046] In certain embodiments, the present disclosure provides compositions and methods of use of oligonucleotides.

[0047] In certain embodiments, the present disclosure provides compositions and methods of use of oligonucleotides capable of inducing both RNA interference and RNase H-mediated knockdown of HSD17B13 target gene RNA. In certain embodiments, the oligonucleotides of such compositions can have the format, structural elements, or base sequences of the oligonucleotides disclosed herein.

[0048] In certain embodiments, an oligonucleotide that induces a particular event or activity is involved in a particular event or activity, such as a decrease in the expression, level, or activity of a target gene or its gene product. In certain embodiments, an oligonucleotide is considered to "induce" a particular event or activity if the presence of the oligonucleotide in a system in which the event or activity may occur correlates with a detectable increase in the occurrence, frequency, intensity, and / or level of the event or activity.

[0049] In certain embodiments, the provided oligonucleotides comprise any one or more structural elements of the oligonucleotides described herein, such as a base sequence (or a portion thereof of at least 15 contiguous bases); an internucleotide linkage pattern (or a portion thereof of at least 5 contiguous internucleotide linkages); an internucleotide linkage stereochemistry pattern (or a portion thereof of at least 5 contiguous internucleotide linkages); a 5'-terminal structure; a 5'-terminal region; a first region; a second region; and a 3'-terminal region (which may be a 3'-terminal dinucleotide and / or a 3'-end cap); and optional additional chemical moieties; in certain embodiments, at least one structural element comprises a chiral center. In certain embodiments, the 3'-terminal dinucleotide may comprise two full nucleotides. In certain embodiments, the oligonucleotide further comprises a chemical moiety selected from, by way of non-limiting example, a targeting moiety, a carbohydrate moiety, a GalNAc moiety, a lipid moiety, and any other chemical moiety described herein or known in the art. In certain embodiments, the APGR-binding moiety is a GalNAc moiety or a variant, derivative, or modified version thereof, as described herein and / or known in the art. In certain embodiments, the oligonucleotide is an RNAi agent. In certain embodiments, the first region is a seed region. In certain embodiments, the second region is a post-seed region.

[0050] In certain embodiments, the provided oligonucleotide comprises any one or more structural elements of the RNAi agent described herein, such as a 5'-terminal structure; a 5'-terminal region; a seed region; a post-seed region (the region between the seed region and the 3'-terminal region); and a 3'-terminal region (which may be a 3'-terminal dinucleotide and / or a 3'-end cap); and an optional additional chemical moiety, wherein in certain embodiments, at least one structural element comprises a chiral center for chiral control. In certain embodiments, the 3'-terminal dinucleotide may comprise two full nucleotides. In certain embodiments, the oligonucleotide further comprises a chemical moiety selected from, by way of non-limiting example, a targeting moiety, a carbohydrate moiety, a GalNAc moiety, and a lipid moiety. In certain embodiments, the APGR-binding moiety is any GalNAc or variant, derivative, or modification thereof, as described herein or known in the art.

[0051] In certain embodiments, the provided oligonucleotides comprise any one or more structural elements of the oligonucleotides described herein, such as a 5'-terminal structure, a 5'-terminal region, a first region, a second region, a 3'-terminal region, and optional additional chemical moieties, wherein at least one structural element comprises a chiral center. In certain embodiments, the oligonucleotide comprises a span of at least 5 full nucleotides without a 2'-modification. In certain embodiments, the oligonucleotide further comprises an additional chemical moiety selected from, by way of non-limiting example, a targeting moiety, a carbohydrate moiety, a GalNAc moiety, and a lipid moiety. In certain embodiments, the provided oligonucleotides are capable of inducing RNA interference. In certain embodiments, the provided oligonucleotides are capable of inducing RNase H-mediated knockdown. In certain embodiments, the provided oligonucleotides are capable of inducing both RNA interference and RNase H-mediated knockdown. In certain embodiments, the first region is a seed region. In certain embodiments, the second region is a post-seed region.

[0052] In certain embodiments, the provided oligonucleotide comprises any one or more structural elements of an RNAi agent, such as a 5'-end structure, a 5'-end region, a seed region, a post-seed region, and a 3'-end region, and optionally additional chemical moieties, wherein at least one structural element comprises a chiral center for chiral control, and in certain embodiments, the oligonucleotide is also capable of inducing RNase H-mediated knockdown of target gene RNA. In certain embodiments, the oligonucleotide comprises a span of at least five total 2'-deoxynucleotides. In certain embodiments, the oligonucleotide further comprises a chemical moiety selected from, by way of non-limiting example, a targeting moiety, a carbohydrate moiety, a GalNAc moiety, a lipid moiety, and any other additional chemical moiety described herein.

[0053] In certain embodiments, the present disclosure demonstrates that the properties of oligonucleotides can be tuned by chemical modification. In certain embodiments, the present disclosure provides an oligonucleotide composition comprising a first plurality of oligonucleotides having a common base sequence and comprising one or more internucleotide linkage, sugar, and / or base modifications. In certain embodiments, the present disclosure provides an oligonucleotide composition capable of inducing RNA interference, comprising a first plurality of oligonucleotides having a common base sequence and comprising one or more internucleotide linkages, and / or one or more sugar, and / or one or more base modifications. In certain embodiments, the oligonucleotide or oligonucleotide composition is also capable of inducing RNase H-mediated knockdown of HSD17B13 target gene RNA. In certain embodiments, the present disclosure demonstrates that the properties of oligonucleotides, such as activity, toxicity, etc., can be tuned by chemical modification of the sugar, nucleobase, and / or internucleotide linkage. In certain embodiments, the present disclosure provides an oligonucleotide composition comprising a first plurality of oligonucleotides having a common base sequence and comprising one or more modified internucleotide linkages (or "non-natural internucleotide linkages"), such as the natural phosphate internucleotide linkage found in natural DNA and RNA (-OP(O)(OH)O-, which is in the salt form at physiological pH (-OP(O)(O-Oligonucleotide compositions are provided that include a plurality of oligonucleotides comprising a linkage that can be utilized in place of (O-), one or more modified sugar moieties, and / or one or more natural phosphate linkages. In certain embodiments, the provided oligonucleotides can include two or more types of modified internucleotide linkages. In certain embodiments, the provided oligonucleotides include a non-negatively charged internucleotide linkage. In certain embodiments, the non-negatively charged internucleotide linkage is a neutral internucleotide linkage. In certain embodiments, the neutral internucleotide linkage includes a cyclic guanidine moiety. Such a moiety is optionally substituted. In certain embodiments, the provided oligonucleotides include a neutral internucleotide linkage and another internucleotide linkage that is not a neutral backbone. In certain embodiments, the provided oligonucleotides include a neutral internucleotide linkage and a phosphorothioate internucleotide linkage. In certain embodiments, the provided oligonucleotide compositions that include a plurality of oligonucleotides are chiral-controlled, wherein the level of the plurality of oligonucleotides in the composition is controlled or predetermined, and the plurality of oligonucleotides share a common stereochemical configuration at one or more chiral internucleotide linkages. For example, in certain embodiments, a plurality of oligonucleotides share a common stereochemical configuration at 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, 35, 40, 45, 50 or more chiral internucleotide linkages, each independently being either Rp or Sp; in certain embodiments, a plurality of oligonucleotides share a common stereochemical configuration at each chiral internucleotide linkage. In certain embodiments, a chiral internucleotide linkage at which a controlled level of oligonucleotides in a composition share a common stereochemical configuration (independently, either Rp or Sp configuration) is referred to as a chiral-controlled internucleotide linkage.In certain embodiments, the modified internucleotide linkage exists predominantly (e.g., at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, etc.; in certain embodiments, at least 30%; in certain embodiments, at least 40%; in certain embodiments, at least 50%; in certain embodiments, at least 60%; in certain embodiments, at least 70%; in certain embodiments, at least 80%; in certain embodiments, at least 90%; in certain embodiments, at least 99%, etc.) in a neutral or cationic form (or an anionic form (e.g., -OP(O)(O). - )-O- (anionic form of the natural phosphate bond), -OP(O)(S -)-O- (the anionic form of a phosphorothioate bond) and other similar non-negatively charged (neutral or cationic) internucleotide linkages. In certain embodiments, the modified internucleotide linkage is a neutral internucleotide linkage in that it exists predominantly in its neutral form at a certain pH. In certain embodiments, the modified internucleotide linkage is a cationic internucleotide linkage in that it exists predominantly in its cationic form at a certain pH. In certain embodiments, the pH is human physiological pH (approximately 7.4). In certain embodiments, the modified internucleotide linkage is a neutral internucleotide linkage in that at least 90% of the internucleotide linkage exists in its neutral form at pH 7.4 in aqueous solution. In certain embodiments, the modified internucleotide linkage is a neutral internucleotide linkage in that at least 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the internucleotide linkage exists in its neutral form in an aqueous solution of the oligonucleotide. In certain embodiments, the percentage is at least 90%. In certain embodiments, the percentage is at least 95%. In certain embodiments, the percentage is at least 99%. In certain embodiments, a non-negatively charged internucleotide linkage, e.g., a neutral internucleotide linkage, when in its neutral form, has no moieties with a pKa of less than 8, 9, 10, 11, 12, 13, or 14. In certain embodiments, the pKa of an internucleotide linkage in the present disclosure can be represented by the pKa of a CH3-internucleotide linkage -CH3 (i.e., two -CH3 groups replace two nucleotide units linked by the internucleotide linkage). Without wishing to be bound by any particular theory, at least in some cases, neutral internucleotide linkages in oligonucleotides can result in improved properties and / or activity, such as improved delivery, improved resistance to exonucleases and endonucleases, improved cellular uptake, improved endosomal escape, and / or improved nuclear uptake, compared to a comparable nucleic acid that does not contain neutral internucleotide linkages.

[0054] In certain embodiments, the non-negatively charged internucleotide linkages are those described in, e.g., U.S. Patent Nos. 9,394,333, 9,744,183, 9,605,019, 9,598,458, 9,982,257, 10,160,969, 10,479,995, U.S. Patent Application Publication No. 2020 / 0056173, U.S. Patent Application Publication No. 2018 / 0216107, U.S. Patent Application Publication No. 2019 / 0127733, U.S. Patent Application Publication No. 10,450,568, U.S. Patent Application Publication No. 2019 / 0077817, U.S. Patent Application Publication No. 2019 / 0249173, U.S. Patent Application Publication No. 2019 / 0259173, U.S. Patent Application Publication No. 2019 / 0269173, U.S. Patent Application Publication No. 2019 / 0279173, U.S. Patent Application Publication No. 2019 / 026 ... and / or WO 2019 / 032612, having a structure of formula In-1, In-2, In-3, II, II-a-1, II-a-2, II-b-1, II-b-2, II-c-1, II-c-2, II-d-1, II-d-2, as described in, for example, WO 2019 / 0375774, WO 2018 / 223056, WO 2018 / 223073, WO 2018 / 223081, WO 2018 / 237194, WO 2019 / 032607, WO 2019 / 055951, WO 2019 / 075357, WO 2019 / 200185, WO 2019 / 217784, and / or WO 2019 / 032612. In certain embodiments, the non-negatively charged internucleotide linkage comprises a cyclic guanidine moiety. In certain embodiments, the modified internucleotide linkage comprising a cyclic guanidine moiety has the following structure: [ka] In certain embodiments, the neutral internucleotide linkage comprising a cyclic guanidine moiety is chiral controlled. In certain embodiments, the present disclosure relates to a composition comprising an oligonucleotide comprising at least one neutral internucleotide linkage and at least one phosphorothioate internucleotide linkage.

[0055] In certain embodiments, the present disclosure relates to a composition comprising an oligonucleotide comprising at least one neutral internucleotide linkage and at least one phosphorothioate internucleotide linkage, wherein the phosphorothioate internucleotide linkage is a chiral internucleotide linkage of the Sp configuration.

[0056] In certain embodiments, the present disclosure relates to a composition comprising an oligonucleotide comprising at least one neutral internucleotide linkage and at least one phosphorothioate internucleotide linkage, wherein the phosphorothioate is a chiral-controlling internucleotide linkage of the Rp configuration.

[0057] In certain embodiments, the present disclosure provides at least one neutral internucleotide linkage comprising a Tmg group. [ka] and a composition comprising an oligonucleotide comprising at least one phosphorothioate.

[0058] In certain embodiments, each internucleotide linkage in the oligonucleotide is independently selected from a natural phosphate linkage, a phosphorothioate linkage, and a non-negatively charged internucleotide linkage (e.g., n001, n003, n004, n006, n008, n009, n013, n020, n021, n025, n026, n029, n031, n037, n046, n047, n048, n054, or n055). In some embodiments, each internucleotide linkage in the oligonucleotide is independently selected from a natural phosphate linkage, a phosphorothioate linkage, and a neutral internucleotide linkage (e.g., n001, n003, n004, n006, n008, n009, n013 n020, n021, n025, n026, n029, n031, n037, n046, n047, n048, n054, or n055).

[0059] In certain embodiments, the present disclosure relates to compositions comprising an oligonucleotide comprising at least one neutral internucleotide linkage comprising a Tmg group and at least one phosphorothioate, wherein the phosphorothioate is a chiral-controlling internucleotide linkage of the Sp configuration.

[0060] In certain embodiments, the present disclosure relates to compositions comprising an oligonucleotide comprising at least one neutral internucleotide linkage selected from neutral internucleotide linkages comprising a Tmg group and at least one phosphorothioate, wherein the phosphorothioate is a chiral-controlling internucleotide linkage of the Rp configuration.

[0061] Various types of internucleotide linkages differ in nature. Without wishing to be bound by any theory, the present disclosure notes that natural phosphate linkages (phosphodiester internucleotide linkages) are anionic and may be unstable in vivo when used alone without other chemical modifications; phosphorothioate internucleotide linkages are anionic, generally more stable in vivo than natural phosphate linkages, and generally more hydrophobic; and neutral internucleotide linkages, such as those exemplified in the present disclosure, that contain cyclic guanidine moieties are neutral at physiological pH and may be more stable in vivo than natural phosphate linkages and more hydrophobic.

[0062] In certain embodiments, the chiral-controlled neutral internucleotide linkage is neutral at physiological pH, chiral-controlled, stable in vivo, hydrophobic, and capable of increasing endosomal escape.

[0063] In certain embodiments, provided oligonucleotides include one or more regions, such as block, wing, core, 5'-end, 3'-end, middle, seed, and post-seed regions. In certain embodiments, the regions (e.g., blocks, wings, cores, 5'-ends, 3'-ends, intermediate regions, etc.) may be selected from the group consisting of those described in, for example, U.S. Patent Nos. 9,394,333, 9,744,183, 9,605,019, 9,598,458, 9,982,257, 10,160,969, 10,479,995, U.S. Patent Application Publication No. 2020 / 0056173, 2018 / 0216107, 2019 / 0127733, 10,450,568, 2019 / 0077817, 2019 / 0249173, and U.S. Patent Application Publication No. 2019 / 0249173. No. 2019 / 0375774, International Publication No. 2018 / 223056, International Publication No. 2018 / 223073, International Publication No. 2018 / 223081, International Publication No. 2018 / 237194, International Publication No. 2019 / 032607, International Publication No. 2019 / 055951, International Publication No. 2019 / 075357, International Publication No. 20 In certain embodiments, the region comprises a non-negatively charged internucleotide linkage, such as formula In-1, In-2, In-3, II, II-a-1, II-a-2, II-b-1, II-b-2, II-c-1, II-c-2, II-d-1, II-d-2, as described in WO 2019 / 200185, WO 2019 / 217784, and / or WO 2019 / 032612. In certain embodiments, the region comprises a neutral internucleotide linkage. In certain embodiments, the region comprises an internucleotide linkage comprising a cyclic guanidine moiety. In certain embodiments, the region comprises an internucleotide linkage having the following structure: [ka] In certain embodiments, such internucleotide linkages are chiral controlled.

[0064] In certain embodiments, the nucleotide is a natural nucleotide. In certain embodiments, the nucleotide is a modified nucleotide. In certain embodiments, the nucleotide is a nucleotide analog. In certain embodiments, the base is a modified base. In certain embodiments, the base is a protected nucleobase, such as a protected nucleobase used in oligonucleotide synthesis. In certain embodiments, the base is a base analog. In certain embodiments, the sugar is a modified sugar. In certain embodiments, the sugar is a sugar analog. In certain embodiments, the internucleotide linkage is a modified internucleotide linkage. In certain embodiments, a nucleotide comprises a base, a sugar, and an internucleotide linkage, wherein each of the base, sugar, and internucleotide linkage is independently and optionally naturally occurring or non-naturally occurring. In certain embodiments, a nucleotide comprises a base and a sugar, wherein each of the base and sugar is independently and optionally naturally occurring or non-naturally occurring. Non-limiting examples of nucleotides include DNA (2'-deoxy) and RNA (2'-OH) nucleotides, as well as those containing one or more modifications in the base, sugar, and / or internucleotide linkage. Non-limiting examples of sugars include ribose and deoxyribose, and ribose and deoxyribose with 2'-modifications, including but not limited to, 2'-F, LNA, 2'-OMe, and 2'-MOE modifications. In certain embodiments, the internucleotide linkage is a non-phosphorus moiety that serves to join two natural or unnatural sugars.

[0065] In certain embodiments, the composition comprises any two or more multimers: a first plurality of oligonucleotides and / or a second plurality of oligonucleotides, wherein the first and second plurality of oligonucleotides are independently capable of independently knocking down the same or different targets by RNA interference and / or RNase H-mediated knockdown.

[0066] In certain embodiments, the present disclosure provides 1) common base sequence; 2) common skeletal bonding patterns; 3) independently having a common stereochemistry at 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, 35, 40, 45, or 50 chiral internucleotide linkages ("chiral-controlling internucleotide linkages"). The present invention provides an oligonucleotide composition comprising a first plurality of oligonucleotides sharing a common structure, wherein the level of the first plurality of oligonucleotides in the composition is chiral-controlled at a predetermined level.

[0067] In certain embodiments, an oligonucleotide composition comprising a plurality of oligonucleotides (e.g., a first plurality of oligonucleotides) is chiral-controlled in that the plurality of oligonucleotides independently share a common stereochemistry at one or more chiral internucleotide linkages. In certain embodiments, the plurality of oligonucleotides independently share a common stereochemistry at 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, 35, 40, 45, 50, or more chiral internucleotide linkages, each of which is independently Rp or Sp. In certain embodiments, the plurality of oligonucleotides share a common stereochemistry at each chiral internucleotide linkage. In certain embodiments, a chiral internucleotide linkage at which a predetermined level of oligonucleotides in the composition share a common stereochemistry (independently Rp or Sp) is referred to as a chiral-controlled internucleotide linkage.

[0068] In certain embodiments, the predetermined level of oligonucleotides of a provided composition, e.g., a first plurality of oligonucleotides of a particular exemplary composition, comprises 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, 35, 40, 45, 50 or more chiral-controlling internucleotide linkages.

[0069] In certain embodiments, at least 5 internucleotide linkages are chiral controlled; in certain embodiments, at least 10 internucleotide linkages are chiral controlled; in certain embodiments, at least 15 internucleotide linkages are chiral controlled; in certain embodiments, each chiral internucleotide linkage is chiral controlled.

[0070] In certain embodiments, 1% to 100% of the chiral internucleotide linkages are chiral controlled, hi certain embodiments, at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the chiral internucleotide linkages are chiral controlled.

[0071] In certain embodiments, the present disclosure provides 1) common base sequence; 2) a common backbone bonding pattern; and 3) Common skeletal chiral center patterns The present invention provides an oligonucleotide composition comprising a first plurality of oligonucleotides sharing a common base sequence and length, a common pattern of backbone linkages, and a common pattern of backbone chiral centers, the composition being a substantially pure preparation of oligonucleotides, wherein a predetermined level of the oligonucleotides in the composition have a common base sequence and length, a common pattern of backbone linkages, and a common pattern of backbone chiral centers. In certain embodiments, the common pattern of backbone chiral centers comprises at least one internucleotide linkage that includes a chiral-controlling chiral center. In certain embodiments, the predetermined level of the oligonucleotides is at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of all oligonucleotides in the provided composition. In certain embodiments, the predetermined level of oligonucleotides is at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of all oligonucleotides in a provided composition that are of or contain a common base sequence. In certain embodiments, all oligonucleotides in a provided composition that are or contain a common base sequence are at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of all oligonucleotides in the composition. In certain embodiments, the predetermined level of oligonucleotides is at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of all oligonucleotides in a provided composition that are or contain a common base sequence, base modification, sugar modification, and / or modified internucleotide linkage.In certain embodiments, all of the oligonucleotides in a provided composition that are or contain a common base sequence, base modification, sugar modification and / or modified internucleotide linkage are at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of all of the oligonucleotides in the composition. In certain embodiments, the predetermined level of oligonucleotides is at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of all oligonucleotides in a provided composition that are or contain a common base sequence, pattern of base modifications, pattern of sugar modifications, and / or pattern of modified internucleotide linkages. In certain embodiments, all of the oligonucleotides in a provided composition that are or contain a common base sequence, pattern of base modifications, pattern of sugar modifications and / or pattern of modified internucleotide linkages are at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of all of the oligonucleotides in the composition. In certain embodiments, the predetermined level of oligonucleotides is at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of all oligonucleotides in a provided composition that share a common base sequence, a common pattern of base modifications, a common pattern of sugar modifications, and / or a common pattern of modified internucleotide linkages.In certain embodiments, all oligonucleotides in a provided composition that share a common base sequence, a common pattern of base modifications, a common pattern of sugar modifications, and / or a common pattern of modified internucleotide linkages represent at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of all oligonucleotides in the composition. In certain embodiments, the predetermined level is between 1 and 100%. In certain embodiments, the predetermined level is at least 1%. In certain embodiments, the predetermined level is at least 5%. In certain embodiments, the predetermined level is at least 10%. In certain embodiments, the predetermined level is at least 20%. In certain embodiments, the predetermined level is at least 30%. In certain embodiments, the predetermined level is at least 40%. In certain embodiments, the predetermined level is at least 50%. In certain embodiments, the predetermined level is at least 60%. In certain embodiments, the predetermined level is at least 10%. In certain embodiments, the predetermined level is at least 70%. In certain embodiments, the predetermined level is at least 80%. In certain embodiments, the predetermined level is at least 90%. In certain embodiments, the predetermined level is at least 5*(½g), where g is the number of chiral-controlling internucleotide linkages. In certain embodiments, the predetermined level is at least 10*(½g), where g is the number of chiral-controlling internucleotide linkages. In certain embodiments, the predetermined level is at least 100*(½g), where g is the number of chiral-controlling internucleotide linkages.In certain embodiments, the predetermined level is at least (0.80) g, where g is the number of chiral-controlling internucleotide linkages. In certain embodiments, the predetermined level is at least (0.80) g, where g is the number of chiral-controlling internucleotide linkages. In certain embodiments, the predetermined level is at least (0.80) g, where g is the number of chiral-controlling internucleotide linkages. In certain embodiments, the predetermined level is at least (0.85) g, where g is the number of chiral-controlling internucleotide linkages. In certain embodiments, the predetermined level is at least (0.90) g, where g is the number of chiral-controlling internucleotide linkages. In certain embodiments, the predetermined level is at least (0.95) g, where g is the number of chiral-controlling internucleotide linkages. In certain embodiments, the predetermined level is at least (0.96) g, where g is the number of chiral-controlling internucleotide linkages. In certain embodiments, the predetermined level is at least (0.97) g, where g is the number of chiral-controlling internucleotide linkages. In certain embodiments, the predetermined level is at least (0.98) g, where g is the number of chiral-controlling internucleotide linkages. In certain embodiments, the predetermined level is at least (0.99) g, where g is the number of chiral-controlling internucleotide linkages. In certain embodiments, to determine the level of oligonucleotides having g chiral-controlling internucleotide linkages in a composition, the product of the diastereopurities of each of the g chiral-controlling internucleotide linkages is used as the level: (diastereopurity of chiral-controlling internucleotide linkage 1) * (diastereopurity of chiral-controlling internucleotide linkage 2) * * (diastereopurity of chiral-controlling internucleotide linkage g).Here, the diastereopurity of each chiral-controlled internucleotide linkage is independently expressed by the diastereopurity of a dimer containing the same internucleotide linkage and nucleosides flanking the internucleotide linkage and prepared in a manner equivalent to the oligonucleotide (e.g., an equivalent or preferably identical oligonucleotide preparation cycle involving equivalent or preferably identical reagents and reaction conditions). In certain embodiments, the level of oligonucleotide and / or diastereopurity can be determined by analytical methods, such as chromatographic methods, spectroscopic methods, spectroscopic methods, or any combination thereof. In particular, the present disclosure encompasses the recognition that stereorandom oligonucleotide preparations contain multiple distinct chemical entities that differ from each other, for example, in the stereochemical structure (or stereochemistry) of individual backbone chiral centers within the oligonucleotide chain. If the stereochemistry of the backbone chiral centers is not controlled, stereorandom oligonucleotide preparations will result in uncontrolled compositions containing an indeterminate level of oligonucleotide stereoisomers. Even though these stereoisomers have the same base sequence and / or chemical modifications, they are different chemical entities due to at least their different backbone stereochemistry, and they may have different properties, such as susceptibility to nucleases, activity, distribution, etc., as demonstrated herein. In certain embodiments, a particular stereoisomer may be defined, for example, by its base sequence, its length, its backbone bond pattern, and its backbone chiral center pattern. In certain embodiments, the present disclosure demonstrates that the improved properties and activity achieved by controlling the stereochemistry within an oligonucleotide can be comparable to or even better than those achieved by the use of chemical modifications.

[0072] In particular, the present disclosure encompasses the recognition that stereorandom oligonucleotide formulations contain multiple distinct chemical entities that differ from one another, for example, in terms of the stereochemical configuration (or stereochemistry) of individual backbone chiral centers within the oligonucleotide chain. If the stereochemistry of the backbone chiral centers is not controlled, stereorandom oligonucleotide formulations result in uncontrolled compositions containing indeterminate levels of oligonucleotide stereoisomers. Even if these stereoisomers have the same base sequence and / or chemical modifications, they are distinct chemical entities due at least to their different backbone stereochemistry, and they may have different properties, such as susceptibility to nucleases, activity, distribution, etc., as demonstrated herein. In certain embodiments, a particular stereoisomer can be defined, for example, by its base sequence, its length, its backbone bond pattern, and its backbone chiral center pattern. In certain embodiments, the present disclosure demonstrates that the improved properties and activity achieved by controlling the stereochemistry within an oligonucleotide can be comparable to or even better than those achieved by the use of chemical modifications.

[0073] In some embodiments, ds oligonucleotides targeting HSD17B13 or compositions of ds oligonucleotides targeting HSD17B13 are useful for preventing or treating HSD17B13-related conditions, disorders, or diseases in a subject in need thereof. In some embodiments, the present disclosure provides methods for preventing or treating HSD17B13-related conditions, disorders, or diseases, comprising administering to a subject suffering from or responding to the same a therapeutically effective amount of a provided oligonucleotide or a pharmaceutical composition that can deliver or contain a therapeutically effective amount of a provided oligonucleotide. In some embodiments, the present disclosure provides pharmaceutical compositions comprising a provided ds oligonucleotide targeting HSD17B13 and a pharmaceutically acceptable carrier. In some embodiments, the oligonucleotide in the pharmaceutical composition is in one or more pharmaceutically acceptable salt forms, such as a sodium salt form, an ammonium salt form, or the like.

[0074] In some embodiments, the oligonucleotide or oligonucleotide composition is useful for the manufacture of a medicament for the prevention or treatment of a condition, disorder or disease associated with HSD17B13, such as NAFLD, NASH, ASH, alcoholic liver disease, non-alcoholic liver disease, alcoholic cirrhosis, non-alcoholic cirrhosis, steatohepatitis, hepatic steatosis, hepatocellular carcinoma, HCV hepatitis, chronic hepatitis, hereditary hemochromatosis, primary sclerosing cholangitis, drug-induced liver injury or hepatocellular necrosis, in a subject in need thereof.

[0075] The provided technology (e.g., oligonucleotides, compositions, methods, etc.) can be used to prevent and / or treat various HSD17B13-related conditions, disorders, or diseases. In some embodiments, the condition, disorder, or disease is NAFLD. In some embodiments, the condition, disorder, or disease is NASH. In some embodiments, the condition, disorder, or disease is ASH. DETAILED DESCRIPTION OF THE INVENTION

[0076] Detailed Description of Specific Embodiments The techniques of the present disclosure may be more readily understood by reference to the following detailed description of specific embodiments.

[0077] definition As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this disclosure, chemical elements are identified according to the Periodic Table of the Elements, CAS system, Handbook of Chemistry and Physics, 75th Edition. Furthermore, general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999 and "March's Advanced Organic Chemistry", 5th Edition, Ed.: Smith, MB and March, J., John Wiley & Sons, New York: 2001.

[0078] As used herein in this disclosure, unless otherwise clear from the context, (i) the term "a" or "an" may be understood to mean "at least one"; (ii) the term "or" may be understood to mean "and / or"; (iii) the terms "comprising," "comprise," "including" (whether or not used in conjunction with "limited to"), and "include" (whether or not used in conjunction with "not limited to") may be understood to encompass the itemized elements or steps, whether presented by themselves or presented with one or more additional elements or steps; (iv) the term "another" may be understood to mean at least an additional / second one or more; (v) the terms "about" and "approximately" may be understood to allow for standard variations as understood by one of ordinary skill in the art; and (vi) when ranges are given, the endpoints are included.

[0079] Unless otherwise specified, descriptions of oligonucleotides and their components (e.g., base sequence, sugar modifications, internucleotide linkages, stereochemistry of linking phosphorus, patterns thereof, etc.) are in the 5' to 3' direction. As one of ordinary skill in the art will understand, in some embodiments, oligonucleotides may be provided and / or utilized as salt forms, particularly pharmaceutically acceptable salt forms, such as sodium salts. Unless otherwise indicated, oligonucleotides include various forms of oligonucleotides. As one of ordinary skill in the art will also understand, in some embodiments, individual oligonucleotides within a composition may be considered to be of the same constitution and / or structure, even within such a composition (e.g., a liquid composition), and certain such oligonucleotides may be in different salt forms at a particular moment (and may be dissolved or may exist as an anionic form when the oligonucleotide chain is, for example, in a liquid composition). For example, one skilled in the art will understand that at a given pH, individual internucleotide bonds along an oligonucleotide chain may be in one of more possible salt forms (e.g., sodium salts or salts of different cations depending on which ions may be present in the preparation or composition) than the acid (H) form, and will understand that these acid forms (e.g., all cations, if present, may be H) may be used. + It will be understood that so long as the nucleotide sequence (replaced by ) is of the same composition and / or structure, such individual oligonucleotides may be considered to be of the same composition and / or structure, as appropriate.

[0080] Analog: The term "analog" includes any chemical moiety that is structurally different from a reference chemical moiety or class of moieties, but that can perform at least one function of such reference chemical moiety or class of moieties. Non-limiting examples include a nucleotide analog that is structurally different from a nucleotide but performs at least one function of a nucleotide, a nucleobase analog that is structurally different from a nucleobase but performs at least one function of a nucleobase, etc.

[0081] Antisense: As used herein, the term "antisense" refers to the characteristic of an oligonucleotide or other nucleic acid having a base sequence complementary or substantially complementary to a target nucleic acid to which it can hybridize. In some embodiments, the target nucleic acid is a target gene mRNA. In some embodiments, hybridization is required for or results in an activity, such as a reduction in the level, expression, or activity of the target nucleic acid or its gene product. The term "antisense oligonucleotide," as used herein, refers to an oligonucleotide that is complementary to a target nucleic acid. In some embodiments, antisense oligonucleotides can induce a reduction in the level, expression, or activity of the target nucleic acid or its gene product. In some embodiments, antisense oligonucleotides can induce a reduction in the level, expression, or activity of the target nucleic acid or its product by a mechanism involving RNA interference.

[0082] Chiral control: As used herein, "chiral control" refers to the control of the stereochemical assignment of the chiral linking phosphorus at a chiral internucleotide linkage within an oligonucleotide. As used herein, a chiral internucleotide linkage is an internucleotide linkage in which the linking phosphorus is chiral. In some embodiments, the control is achieved through chiral elements not present in the sugar and base portions of the oligonucleotide; for example, in some embodiments, the control is achieved by using one or more chiral auxiliary groups during the preparation of the oligonucleotide, as described herein, which chiral auxiliary groups are often part of the chiral phosphoramidites used during the oligonucleotide preparation. In contrast to chiral control, those skilled in the art understand that conventional oligonucleotide synthesis without the use of chiral auxiliary groups cannot control the stereochemistry at the chiral internucleotide linkage when such conventional oligonucleotide synthesis is used to form the chiral internucleotide linkage. In some embodiments, the stereochemical assignment of each chiral linking phosphorus at each chiral internucleotide linkage within an oligonucleotide is controlled.

[0083] Chirality-controlled oligonucleotide composition: As used herein, the terms "chirality-controlled oligonucleotide composition," "chirality-controlled nucleic acid composition," and the like refer to a composition comprising multiple oligonucleotides (or nucleic acids) that share 1) a common base sequence, 2) a common backbone linkage pattern, and 3) a common backbone phosphorus modification pattern, where the multiple oligonucleotides (or nucleic acids) share the same linkage phosphorus stereochemistry at one or more chiral internucleotide linkages (chirality-controlled or sterically-restricted internucleotide linkages where the chiral linkage phosphorus is Rp or Sp in the composition ("sterically restricted"), rather than a random Rp and Sp mixture as in the case of non-chirality-controlled internucleotide linkages). The level of multiple oligonucleotides (or nucleic acids) in the chirality-controlled oligonucleotide composition is predetermined / controlled (e.g., by chirality-controlled oligonucleotide preparation that stereoselectively forms one or more chiral internucleotide linkages). In some embodiments, about 1% to 100% (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 50%, 50%, 60%, 70%, 80%, 90%, 95 ... %, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%) are a plurality of oligonucleotides.In some embodiments, about 1% to 100% (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95 ... 90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%) are a plurality of oligonucleotides. In some embodiments, the level is between about 1% and 100% (e.g., between about 5% and 100%) of all oligonucleotides in a composition, or of all oligonucleotides in a composition (e.g., of multiple oligonucleotides or types of oligonucleotides) that share a common base sequence, or of all oligonucleotides in a composition that share a common base sequence, a common backbone linkage pattern, and a common backbone phosphorus modification pattern, or of all oligonucleotides in a composition that share a common base sequence, a common base modification pattern, a common sugar modification pattern, a common internucleotide linkage type pattern, and / or a common internucleotide linkage modification pattern. %, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80 to 100%, 90 to 100%, 95 to 100%, 50% to 90%, or approximately 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 9 0%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%.In some embodiments, the plurality of oligonucleotides share the same stereochemistry at about 1-50 chiral internucleotide linkages. In some embodiments, the plurality of oligonucleotides share the same stereochemistry at about 1%-100% of the chiral internucleotide linkages. In some embodiments, the plurality of oligonucleotides (or nucleic acids) are of the same chemical constitution (as one of skill in the art will understand, some embodiments may exist in one or more forms, e.g., acid form, salt form, etc.). In some embodiments, the level of the plurality of oligonucleotides (or nucleic acids) is about 1%-100% of all oligonucleotides (or nucleic acids) in the composition that share the same constitution as the plurality of oligonucleotides (or nucleic acids). In some embodiments, each chiral internucleotide linkage is a chiral controlled internucleotide linkage, and the composition is a completely chiral controlled oligonucleotide composition. In some embodiments, the plurality of oligonucleotides (or nucleic acids) are structurally identical. In some embodiments, the chiral-controlled internucleotide linkage has a diastereomeric purity of at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%, typically at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%. In some embodiments, the chiral-controlled internucleotide linkage has a diastereomeric purity of at least 95%. In some embodiments, the chiral-controlled internucleotide linkage has a diastereomeric purity of at least 96%. In some embodiments, the chiral-controlled internucleotide linkage has a diastereomeric purity of at least 97%. In some embodiments, the chiral-controlled internucleotide linkage has a diastereomeric purity of at least 98%. In some embodiments, the chiral-controlled internucleotide linkage has a diastereomeric purity of at least 99%. In some embodiments, the percentages (e.g., levels described herein) are (DS). nc or at least (DS) ncwhere DS is the diastereomeric purity as described herein (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% or greater), and nc is the number of chiral-controlling internucleotide linkages as described herein (e.g., 1 to 50, 1 to 40, 1 to 30, 1 to 25, 1 to 20, 5 to 50, 5 to 40, 5 to 30, 5 to 25, 5 to 20, 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 greater). In some embodiments, the percentage (e.g., at a level described herein) is (DS) nc or at least (DS) nc where DS is between 95% and 100%. For example, if DS is 99% and nc is 10, the percentage is 90% or at least 90% (99%). 10≈0.90=90%). In some embodiments, the level of multiple oligonucleotides in a composition is expressed as the product of the diastereopurities of each chiral-controlled internucleotide bond in the oligonucleotide. In some embodiments, the diastereopurity of an internucleotide bond linking two nucleosides in an oligonucleotide (or nucleic acid) is expressed by the diastereopurity of the internucleotide bond of a dimer linking the same two nucleosides, where the dimer is prepared using equivalent conditions, in some instances, identical synthesis cycle conditions (e.g., in a bond between Nx and Ny in an oligonucleotide...NxNy..., the dimer is NxNy). In some embodiments, not all chiral internucleotide bonds are chiral-controlled internucleotide bonds, and the composition is a partially chiral-controlled oligonucleotide composition. In some embodiments, the non-chirally controlled internucleotide linkages have a diastereomeric purity of less than about 80%, 75%, 70%, 65%, 60%, 55%, or about 50%, as typically observed in stereorandom oligonucleotide compositions (e.g., those resulting from conventional oligonucleotide synthesis, e.g., phosphoramidite methods, as will be understood by those skilled in the art). In some embodiments, the multiple oligonucleotides (or nucleic acids) are of the same type. In some embodiments, the chirality-controlled oligonucleotide composition comprises non-random or controlled levels of individual oligonucleotide or nucleic acid types. For example, in some embodiments, the chirality-controlled oligonucleotide composition comprises no more than one oligonucleotide type. In some embodiments, the chirality-controlled oligonucleotide composition comprises two or more oligonucleotide types. In some embodiments, the chirality-controlled oligonucleotide composition comprises multiple oligonucleotide types. In some embodiments, the chirality-controlled oligonucleotide composition is a composition of oligonucleotides of a certain oligonucleotide type, and the composition comprises non-random or controlled levels of multiple oligonucleotides of that oligonucleotide type.

[0084] Internucleotide linkage: As used herein, the phrase "internucleotide linkage" generally refers to the bond connecting the nucleoside units of an oligonucleotide or nucleic acid. In some embodiments, the internucleotide linkage is a phosphodiester bond (a natural phosphate bond (-OP(=O)(OH)O-), which may exist as a salt form, as will be understood by those skilled in the art) that is widely found in naturally occurring DNA and RNA molecules. In some embodiments, the internucleotide linkage is a modified internucleotide linkage (not a natural phosphate bond). In some embodiments, the internucleotide linkage is a "modified internucleotide linkage" in which at least one oxygen atom or -OH of the phosphodiester bond is replaced with a different organic or inorganic moiety. In some embodiments, such organic or inorganic moiety is selected from =S, =Se, =NR', -SR', -SeR', -N(R')2, B(R')3, -S-, -Se-, and -N(R')-, where each R' is independently as defined and described in this disclosure. In some embodiments, the internucleotide linkage is a phosphotriester linkage, a phosphorothioate linkage (or a phosphorothioate diester linkage, -OP(=O)(SH)O-, which may exist as a salt form as will be understood by those skilled in the art), or a phosphorothioate triester linkage. In some embodiments, the modified internucleotide linkage is a phosphorothioate linkage. In some embodiments, the internucleotide linkage is, for example, one of a PNA (peptide nucleic acid) or PMO (phosphorodiamidate morpholino oligomer) linkage. In some embodiments, the modified internucleotide linkage is a non-negatively charged internucleotide linkage. In some embodiments, the modified internucleotide linkage is a neutral internucleotide linkage (e.g., n001 in certain provided oligonucleotides). It will be understood by those skilled in the art that an internucleotide linkage can exist as an anion or cation at a given pH due to the presence of an acid or base moiety in the linkage.In some embodiments, the modified internucleotide linkages are those designated s, s1, s2, s3, s4, s5, s6, s7, s8, s9, s10, s11, s12, s13, s14, s15, s16, s17, and s18 as described in WO 2017 / 210647.

[0085] In vitro: As used herein, the term "in vitro" refers to events that take place not within a living organism (e.g., an animal, plant, and / or microorganism), but in an artificial environment, such as a test tube or reaction vessel, cell culture, etc.

[0086] In vivo: As used herein, the term "in vivo" refers to events that take place within an organism (e.g., an animal, a plant, and / or a microorganism).

[0087] Bound phosphorus: As defined herein, the phrase "bound phosphorus" is used to indicate that the particular phosphorus atom being referenced is a phosphorus atom present in an internucleotide linkage, which corresponds to the phosphorus atom of a phosphodiester internucleotide linkage present in naturally occurring DNA and RNA. In some embodiments, the bound phosphorus atom is present in a modified internucleotide linkage, wherein each oxygen atom of the phosphodiester linkage is optionally and independently replaced by an organic or inorganic moiety. In some embodiments, the bound phosphorus atom is P of Formula I described herein. In some embodiments, the bound phosphorus atom is chiral. In some embodiments, the bound phosphorus atom is achiral (e.g., like a natural phosphate linkage).

[0088] Linker: The terms "linker," "linker moiety," and the like refer to any chemical moiety that links one chemical moiety to another. As will be understood by one of skill in the art, a linker can be bivalent, trivalent, or higher, depending on the number of chemical moieties it connects. In some embodiments, a linker is a moiety that links one oligonucleotide to another in a multimer. In some embodiments, a linker is a moiety that is optionally located between the terminal nucleoside and a solid support, or between the terminal nucleoside and another nucleoside, nucleotide, or nucleic acid. In some embodiments, a linker links a chemical moiety (e.g., a targeting moiety, a lipid moiety, a carbohydrate moiety, etc.) to the oligonucleotide chain (e.g., via its 5' end, 3' end, nucleobase, sugar, internucleotide linkage, etc.) in an oligonucleotide.

[0089] Modified nucleobase: The terms "modified nucleobase," "modified base," and the like refer to a chemical moiety that is chemically different from a nucleobase but can perform at least one function of a nucleobase. In some embodiments, a modified nucleobase is a nucleobase that includes a modification. In some embodiments, a modified nucleobase can perform at least one function of a nucleobase, for example, forming a moiety in a polymer that can base pair with a nucleic acid that contains at least a complementary sequence of bases. In some embodiments, a modified nucleobase is a substituted A, T, C, G, or U, or a substituted tautomer of A, T, C, G, or U. In some embodiments, a modified nucleobase in the context of an oligonucleotide refers to a nucleobase that is not A, T, C, G, or U.

[0090] Modified nucleoside: The term "modified nucleoside" refers to a moiety derived from or chemically similar to a natural nucleoside, but containing a chemical modification that distinguishes it from a natural nucleoside. Non-limiting examples of modified nucleosides include those containing modifications at the base and / or sugar. Non-limiting examples of modified nucleosides include those having a 2' modification at the sugar. Non-limiting examples of modified nucleosides also include abasic nucleosides (lacking a nucleobase). In some embodiments, a modified nucleoside can perform at least one function of a nucleoside, e.g., form a moiety in a polymer capable of base pairing with a nucleic acid containing at least a complementary sequence of bases.

[0091] Modified Nucleotide: The term "modified nucleotide" includes any chemical moiety that is structurally different from a naturally occurring nucleotide but can perform at least one function of a naturally occurring nucleotide. In some embodiments, a modified nucleotide comprises a modification in the sugar, base, and / or internucleotide linkage. In some embodiments, a modified nucleotide comprises a modified sugar, modified nucleobase, and / or modified internucleotide linkage. In some embodiments, a modified nucleotide can perform at least one function of a nucleotide, e.g., to form a subunit in a polymer capable of base pairing with a nucleic acid comprising at least a complementary sequence of bases.

[0092] Modified sugar: The term "modified sugar" refers to a moiety that can replace a sugar. The modified sugar mimics the spatial arrangement, electronic properties, or some other physicochemical property of the sugar. In some embodiments, as described in this disclosure, the modified sugar is a substituted ribose or deoxyribose. In some embodiments, the modified sugar comprises a 2'-modification. Examples of useful 2'-modifications are widely available in the art and described herein. In some embodiments, the 2'-modification is a 2'-OR (where R is an optionally substituted C 1~10In some embodiments, the 2'-modification is 2'-OMe. In some embodiments, the 2'-modification is 2'-MOE. In some embodiments, the modified sugar is a bicyclic sugar (e.g., sugars used in LNA, BNA, etc.). In some embodiments, in the context of oligonucleotides, the modified sugar is a sugar that is not ribose or deoxyribose, as typically found in natural RNA or DNA.

[0093] Nucleic Acid: As used herein, the term "nucleic acid" includes any nucleotide and polymers thereof. The term "polynucleotide," as used herein, refers to a polymeric form of nucleotides of any length, either ribonucleotides (RNA) or deoxyribonucleotides (DNA), or a combination thereof. These terms refer to the primary structure of the molecule and thus include double- and single-stranded DNA and double- and single-stranded RNA. These terms include, by equivalents, analogs of RNA or DNA, including modified nucleotides and / or modified polynucleotides, such as, but not limited to, through methylated, protected, and / or capped nucleotides or polynucleotides. These terms encompass poly- or oligo-ribonucleotides (RNA) and poly- or oligo-deoxyribonucleotides (DNA); RNA or DNA derived from nucleobases and / or N- or C-glycosides of modified nucleobases; nucleic acids derived from sugars and / or modified sugars; and nucleic acids derived from phosphate bridges and / or modified internucleotide linkages. The term encompasses nucleic acids containing any combination of nucleobases, modified nucleobases, sugars, modified sugars, phosphate bridges, or modified internucleotide linkages. Examples include, but are not limited to, nucleic acids containing ribose moieties, nucleic acids containing deoxyribose, nucleic acids containing both ribose and deoxyribose moieties, and nucleic acids containing ribose and modified ribose moieties. Unless otherwise specified, the prefix poly- refers to nucleic acids containing from 2 to about 10,000 nucleotide monomer units, and the prefix oligo- refers to nucleic acids containing from 2 to about 200 nucleotide monomer units.

[0094] Nucleobase: The term "nucleobase" refers to the portion of a nucleic acid that participates in hydrogen bonding to bind one nucleic acid strand to another complementary strand in a sequence-specific manner. The most common naturally occurring nucleobases are adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T). In some embodiments, naturally occurring nucleobases are modified adenine, guanine, uracil, cytosine, or thymine. In some embodiments, naturally occurring nucleobases are methylated adenine, guanine, uracil, cytosine, or thymine. In some embodiments, a nucleobase comprises a heteroaryl ring (where the ring atom is nitrogen), and in a nucleoside, the nitrogen is attached to the sugar moiety. In some embodiments, a nucleobase comprises a heterocyclic ring (where the ring atom is nitrogen), and in a nucleoside, the nitrogen is attached to the sugar moiety. In some embodiments, the nucleobase is a nucleobase other than the "modified nucleobases" adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T). In some embodiments, the modified nucleobase is a substituted A, T, C, G, or U. In some embodiments, the modified nucleobase is a substituted tautomer of A, T, C, G, or U. In some embodiments, the modified nucleobase is a methylated adenine, guanine, uracil, cytosine, or thymine. In some embodiments, the modified nucleobase mimics the spatial arrangement, electronic properties, or some other physicochemical property of the nucleobase and retains the hydrogen bonding properties that bind one nucleic acid strand to another in a sequence-specific manner. In some embodiments, the modified nucleobase can pair with all five naturally occurring bases (uracil, thymine, adenine, cytosine, or guanine) without substantially affecting the melting behavior or recognition by intracellular enzymes or activities of the oligonucleotide duplex. As used herein, the term "nucleobase" also encompasses structural analogs that are used in place of natural or naturally occurring nucleotides, such as modified nucleobases and nucleobase analogs. In some embodiments, the nucleobase is an optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U.In some embodiments, "nucleobase" refers to a nucleobase unit in an oligonucleotide or nucleic acid (e.g., A, T, C, G, or U as in an oligonucleotide or nucleic acid).

[0095] Nucleoside: The term "nucleoside" refers to a moiety in which a nucleobase or modified nucleobase is covalently linked to a sugar or modified sugar. In some embodiments, the nucleoside is a natural nucleoside, such as adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, or deoxycytidine. In some embodiments, the nucleoside is a modified nucleoside, such as a substituted natural nucleoside selected from adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, and deoxycytidine. In some embodiments, the nucleoside is a modified nucleoside, such as a substituted tautomer of a natural nucleoside selected from adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, and deoxycytidine. In some embodiments, "nucleoside" refers to a nucleoside unit in an oligonucleotide or nucleic acid.

[0096] Nucleotide: As used herein, the term "nucleotide" refers to a monomeric unit of a polynucleotide consisting of a nucleobase, a sugar, and one or more internucleotide linkages (e.g., phosphate linkages in natural DNA and RNA). Naturally occurring bases [guanine (G), adenine (A), cytosine (C), thymine (T), and uracil (U)] are derivatives of purines or pyrimidines, but should be understood to include both naturally occurring and non-naturally occurring base analogs. Naturally occurring sugars are the pentose (five-carbon sugars) deoxyribose (forming DNA) or ribose (forming RNA), but should be understood to include both naturally occurring and non-naturally occurring sugar analogs. Nucleotides are linked via internucleotide linkages to form nucleic acids or polynucleotides. Many internucleotide linkages are known in the art (e.g., but not limited to, phosphate, phosphorothioate, boranophosphate, etc.). Artificial nucleic acids include PNA (peptide nucleic acid), phosphotriester, phosphorothioate, H-phosphonate, phosphoramidate, boranophosphate, methylphosphonate, phosphonoacetate, thiophosphonoacetate, and other variants of the phosphate backbone of natural nucleic acids, such as those described herein. In some embodiments, natural nucleotides contain naturally occurring bases, sugars, and internucleotide linkages. As used herein, the term "nucleotide" also encompasses structural analogs that are used in place of natural or naturally occurring nucleotides, such as modified nucleotides and nucleotide analogs. In some embodiments, "nucleotide" refers to a nucleotide unit in an oligonucleotide or nucleic acid.

[0097] Oligonucleotide: The term "oligonucleotide" refers to a polymer or oligomer of nucleotides, which may contain any combination of natural and unnatural nucleobases, sugars, and internucleotide linkages.

[0098] Oligonucleotides can be single-stranded or double-stranded. Single-stranded oligonucleotides can have a double-stranded region (formed by two portions of the single-stranded oligonucleotide), and double-stranded oligonucleotides containing two oligonucleotide strands can have a single-stranded region, for example, in the region where the two oligonucleotide strands are not complementary to each other. Examples of oligonucleotides include, but are not limited to, structural genes, genes including regulatory and termination regions, self-replicating systems such as viruses or plasmid DNA, single-stranded and double-stranded RNAi agents and other RNA interference reagents (RNAi agents or iRNA agents), shRNA, antisense oligonucleotides, ribozymes, microRNAs, microRNA mimics, supermirs, aptamers, antimirs, antagomirs, Ul adapters, triplex-forming oligonucleotides, G-quadruplex oligonucleotides, RNA activators, immunostimulatory oligonucleotides, and decoy oligonucleotides.

[0099] Oligonucleotides of the present disclosure can be of various lengths. In certain embodiments, oligonucleotides can range from about 2 to about 200 nucleosides in length. In various related embodiments, single-stranded, double-stranded, or triple-stranded oligonucleotides can range in length from about 4 to about 10 nucleosides, about 10 to about 50 nucleosides, about 20 to about 50 nucleosides, about 15 to about 30 nucleosides, or about 20 to about 30 nucleosides in length. In some embodiments, oligonucleotides are about 9 to about 39 nucleosides in length. In some embodiments, oligonucleotides are at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleosides in length. In some embodiments, oligonucleotides are at least 19 nucleosides in length. In some embodiments, the oligonucleotide is at least 20 nucleosides in length. In some embodiments, the oligonucleotide is at least 25 nucleosides in length. In some embodiments, the oligonucleotide is at least 30 nucleosides in length. In some embodiments, the oligonucleotide is a duplex of complementary strands at least 18 nucleosides in length. In some embodiments, the oligonucleotide is a duplex of complementary strands at least 21 nucleosides in length. In some embodiments, each nucleoside counted in the length of the oligonucleotide independently comprises A, T, C, G, or U, or an optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U.

[0100] Oligonucleotide type: As used herein, the phrase "oligonucleotide type" refers to an oligonucleotide having a particular base sequence, backbone linkage pattern (i.e., pattern of internucleotide linkage types, e.g., phosphate, phosphorothioate, phosphorothioate triester, etc.), pattern of backbone chiral centers [i.e., pattern of bond phosphorus stereochemistry (Rp / Sp)], and pattern of backbone phosphorus modifications (e.g., "-XLR" of Formula I described herein). 1"Type" is used to define oligonucleotides having a pattern of "types" of groups. In some embodiments, oligonucleotides of a generically named "type" are structurally identical to each other.

[0101] Those skilled in the art will appreciate that the synthetic methods of the present disclosure provide a degree of control during the synthesis of an oligonucleotide chain, such that each nucleotide unit of the oligonucleotide chain can be designed and / or selected in advance to have a specific stereochemistry at the linking phosphorus and / or a specific modification at the linking phosphorus, and / or a specific base and / or a specific sugar. In some embodiments, the oligonucleotide chain is designed and / or selected in advance to have a specific combination of stereocenters at the linking phosphorus. In some embodiments, the oligonucleotide chain is designed and / or determined to have a specific combination of modifications at the linking phosphorus. In some embodiments, the oligonucleotide chain is designed and / or selected to have a specific combination of bases. In some embodiments, the oligonucleotide chain is designed and / or selected to have a specific combination of one or more of the above structural features. In some embodiments, the present disclosure provides compositions (e.g., chiral-controlled oligonucleotide compositions) comprising or consisting of a plurality of oligonucleotide molecules. In some embodiments, all such molecules are of the same type (i.e., structurally identical to one another). However, in some embodiments, the provided compositions typically comprise multiple oligonucleotides of different types in predetermined relative amounts.

[0102] Optionally substituted: As described herein, compounds of the present disclosure, such as oligonucleotides, may contain optionally substituted and / or substituted moieties. In general, the term "substituted," whether preceded by the term "optionally," means that one or more hydrogens of the specified moiety have been replaced with a suitable substituent. Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and if more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents may be the same or different at all positions. In some embodiments, an optionally substituted group is unsubstituted. Combinations of substituents envisioned by the present disclosure preferably result in the formation of stable or chemically feasible compounds. As used herein, the term "stable" refers to compounds that remain substantially unchanged when subjected to conditions that allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein. Specific substituents are described below.

[0103] Suitable monovalent substituents on substitutable atoms, e.g., suitable carbon atoms, are independently halogen; -(CH) 0~4 R°;-(CH2) 0~4 OR°;-O(CH2) 0~4 R°, -O-(CH2) 0~4 C(O)OR°;-(CH2) 0~4 CH(OR°)2; R° can be substituted with -(CH2) 0~4 Ph; R° may be substituted with -(CH2) 0~4 O(CH2) 0~1 Ph; -CH=CHPh, which may be substituted with R°; -(CH2), which may be substituted with R° 0~4 O(CH2) 0~1 -pyridyl; -NO2; -CN; -N3; ​​(CH2) 0~4 N(R°)2;-(CH2) 0~4 N(R°)C(O)R°;-N(R°)C(S)R°;-(CH2) 0~4N(R°)C(O)NR°2; -N(R°)C(S)NR°2; -(CH2) 0~4 N(R°)C(O)OR°; -N(R°)N(R°)C(O)R°; -N(R°)N(R°)C(O)NR°2; -N(R°)N(R°)C(O)OR°; -(CH2) 0~4 C(O)R°; -C(S)R°; -(CH2) 0~4 C(O)OR°; -(CH2) 0~4 C(O)SR°; -(CH2) 0~4 C(O)OSiR°3; -(CH2) 0~4 OC(O)R°; -OC(O)(CH2) 0~4 SR°、 -SC(S)SR°; -(CH2) 0~4 SC(O)R°; -(CH2) 0~4 C(O)NR°2; -C(S)NR°2; -C(S)SR°; (CH2) 0~4 OC(O)NR°2; C(O)N(OR°)R°; -C(O)C(O)R°; -C(O)CH2C(O)R°; -C(NOR°)R°; (CH2) 0~4 SSR°; -(CH2) 0~4 S(O)2R°; -(CH2) 0~4 S(O)2OR°; -(CH2) 0~4 OS(O)2R°; -S(O)2NR°2; (CH2) 0~4 S(O)R°; -N(R°)S(O)2NR°2; -N(R°)S(O)2R°; -N(OR°)R°; -C(NH)NR°2; -Si(R°)3; -OSi(R°)3; -B(R°)2; -OB(R°)2; -OB(OR°)2; -P(R°)2; -P(OR°)2; -P(R°)(OR°); -OP(R°)2; -OP(OR°)2; -OP(R°)(OR°); -P(O)(R°)2; -P(O)(OR°)2; -OP(O)(R°)2; -OP(O)(OR°)2; -OP(O)(OR°)(SR°); -SP(O)(R°)2; -SP(O)(OR°)2; -N(R°)P(O)(R°)2; -N(R°)P(O)(OR°)2; -P(R°)2[B(R°)3]; -P(OR°)2[B(R°)3]; -OP(R°)2[B(R°)3]; -OP(OR°)2[B(R°)3]; -(C 1~4 linear or branched alkylene)O-N(R°)2; or -(C 1~4linear or branched alkylene)C(O)ON(R°)2, where each R° may be substituted as defined herein and independently represents hydrogen, C 1~20 C having 1 to 5 heteroatoms independently selected from aliphatic, nitrogen, oxygen, sulfur, silicon, and phosphorus 1~20 Heteroaliphatic, -CH2-(C 6~14 aryl), -O(CH2) 0~1 (C 6~14 aryl), -CH2- (a 5-14 membered heteroaryl ring), a 5-20 membered monocyclic, bicyclic or polycyclic saturated, partially unsaturated or aryl ring having 0-5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon and phosphorus, or notwithstanding the above definitions, two independent occurrences of R° taken together with their intervening atoms form a 5-20 membered monocyclic, bicyclic or polycyclic saturated, partially unsaturated or aryl ring having 0-5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon and phosphorus, which may be substituted as defined below.

[0104] Suitable monovalent substituents on R° (or the ring formed by two independent occurrences of R° taken together with their intervening atoms) are independently halogen, —(CH) 0~2 R ● ,-(Halo R ● ), -(CH2) 0~2 OH, -(CH2) 0~2 OR ● , -(CH2) 0~2 CH(OR ● )2;-O(HaloR ● ), -CN, -N3, -(CH2) 0~2 C(O)R ● , -(CH2) 0~2 C(O)OH, -(CH2) 0~2 C(O)OR ● , -(CH2) 0~2 SR ● , -(CH2) 0~2 SH, -(CH2) 0-2 NH2, -(CH2) 0~2 NHR ● , -(CH2) 0~2 NR ●2, -NO2, -SiR ● 3. -OSiR ● 3. -C(O)SR ● 、 -(C 1~4 Linear or branched alkylene)C(O)OR ● , or -SSR ● where each R ● is unsubstituted or, when preceded by "halo", is substituted with one or more halogens only, and independently represents C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0~1 Ph and a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. Suitable divalent substituents on a saturated carbon atom of R° include =0 and =S.

[0105] For example, suitable divalent substituents on suitable carbon atoms are, independently, the following: =O, =S, =NNR * 2, =NNHC(O)R * , =NNHC(O)OR * , =NNHS(O)2R * , =NR * , =NOR * , -O(C(R * 2)) 2~3 O- or -S(C(R * 2)) 2~3 S-, where R * Each independent occurrence of is selected from hydrogen, C which may be substituted as defined below 1~6 A 5- to 6-membered unsubstituted saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from aliphatic, nitrogen, oxygen, and sulfur. Suitable divalent substituents attached to adjacent substitutable carbon atoms of an "optionally substituted" group include -O(CR * 2) 2~3 O-, where R * Each independent occurrence of is selected from hydrogen, C which may be substituted as defined below 1~6It is selected from aliphatic and unsubstituted 5-6 membered saturated, partially unsaturated, and aryl rings having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0106] R * Suitable substituents on the aliphatic group are independently halogen, —R ● ,-(Halo R ● ), -OH, -OR ● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2 or -NO2, where each R ● is unsubstituted or, when preceded by "halo", is substituted with one or more halogens only, and independently represents C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0~1 Ph or a 5-6 membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen and sulfur.

[0107] In some embodiments, suitable substituents on a substitutable nitrogen are independently —R † , -NR † 2. -C(O)R † , -C(O)OR † , -C(O)C(O)R † , -C(O)CHC(O)R † , -S(O)2R † , -S(O)NR † 2. -C(S)NR † 2. -C(NH)NR † 2 or -N(R † )S(O)2R † where each R † are independently hydrogen, C which may be substituted as defined below 1~6 an aliphatic, unsubstituted -OPh, or an unsubstituted 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or, notwithstanding the above definitions, R †two independent occurrences of, taken together with their intervening atoms, form an unsubstituted 3-12 membered saturated, partially unsaturated or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen and sulfur.

[0108] R † Suitable substituents on the aliphatic group are independently halogen, —R ● ,-(Halo R ● ), -OH, -OR ● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2 or -NO2, where each R ● is unsubstituted or, when preceded by "halo", is substituted with one or more halogens only, and independently represents C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0~1 Ph or a 5-6 membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen and sulfur.

[0109] Oral: The phrases "oral administration" and "orally administered" as used herein have their art-understood meaning and refer to administration of a compound or composition by mouth.

[0110] Parenteral: The phrases "parenteral administration" and "parenterally administered," as used herein, have their art-recognized meaning and refer to modes of administration other than enteral and topical administration, usually by injection, and include, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0111] Partially unsaturated: As used herein, the term "partially unsaturated" refers to a ring moiety that includes at least one double or triple bond. The term "partially unsaturated" is intended to encompass rings with multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties, as defined herein.

[0112] Pharmaceutical composition: As used herein, the term "pharmaceutical composition" refers to an active agent formulated with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in a unit dose suitable for administration in a treatment regimen that exhibits a statistically significant likelihood of achieving a predetermined therapeutic effect when administered to an appropriate population. In some embodiments, the pharmaceutical composition can be specially formulated for administration in solid or liquid form, including those compatible with: oral administration, e.g., drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., those targeting buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, e.g., as a sterile solution or suspension or sustained-release formulation, e.g., by subcutaneous, intramuscular, intravenous, or epidural injection; topical application, e.g., as a cream, ointment, or controlled-release patch or spray applied to the skin, lungs, or oral cavity; vaginal or rectal application, e.g., as a suppository, cream, or foam; sublingual; ocular; transdermal; or intranasal, pulmonary, and other mucosal surface application.

[0113] Pharmaceutically acceptable: As used herein, the phrase "pharmaceutically acceptable" refers to compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0114] Pharmaceutically acceptable carrier: As used herein, the term "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent that encapsulates a material, that is involved in carrying or transporting a compound of interest from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; pH buffer solutions; polyesters, polycarbonates, and / or polyanhydrides; and other non-toxic, compatible substances utilized in pharmaceutical formulations.

[0115] Pharmaceutically acceptable salts: As used herein, the term "pharmaceutically acceptable salts" refers to salts of such compounds that are suitable for use in a pharmaceutical context, i.e., salts that are suitable for use in contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic response, and the like, within the scope of sound medical judgment, commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. provide a detailed description of pharmaceutically acceptable salts in J. Pharmaceutical Sciences, 66: 1-19 (1977). In some embodiments, pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts of amino groups formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or formed by other methods used in the art, such as ion exchange. In some embodiments, pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, and 2-hydroxy-ethanesulfonate. , lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like.In some embodiments, provided compounds contain one or more acidic groups (e.g., oligonucleotides), and pharmaceutically acceptable salts are alkali metal salts, alkaline earth metal salts, or ammonium salts (e.g., ammonium salts of N(R)3, where each R is independently defined and described in this disclosure). Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. In some embodiments, the pharmaceutically acceptable salt is a sodium salt. In some embodiments, the pharmaceutically acceptable salt is a potassium salt. In some embodiments, the pharmaceutically acceptable salt is a calcium salt. In some embodiments, pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyls having 1 to 6 carbon atoms, sulfonates, and arylsulfonates, as appropriate. In some embodiments, provided compounds contain two or more acid groups; for example, oligonucleotides may contain two or more acid groups (e.g., in natural phosphate linkages and / or modified internucleotide linkages). In some embodiments, pharmaceutically acceptable salts, or salts in general, of such compounds contain two or more cations, which may be the same or different. In some embodiments, in pharmaceutically acceptable salts (or salts in general), all ionizable hydrogens in the acid groups (e.g., in aqueous solutions with a pKa of about 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2 or less, in some embodiments, about 7 or less; in some embodiments, about 6 or less; in some embodiments, about 5 or less; in some embodiments, about 4 or less; in some embodiments, about 3 or less) are replaced with cations. In some embodiments, each phosphorothioate and phosphate group independently exists in its salt form (e.g., in the case of a sodium salt, -OP(O)(SNa)-O- and -OP(O)(ONa)-O-, respectively).In some embodiments, each phosphorothioate and phosphate internucleotide linkage is independently present in its salt form (e.g., in the case of sodium salts, -OP(O)(SNa)-O- and -OP(O)(ONa)-O-, respectively). In some embodiments, the pharmaceutically acceptable salt is a sodium salt of an oligonucleotide. In some embodiments, the pharmaceutically acceptable salt is a sodium salt of an oligonucleotide, and each acidic phosphate group and modified phosphate group (e.g., phosphorothioate, phosphate, etc.), if present, is present as a salt form (all sodium salts).

[0116] Protecting Group: The term "protecting group" as used herein refers to a group that is well known in the art and is described in Organic Synthesis, TW Greene and PGM Wuts, 3 rd edition, John Wiley & Sons, 1999. Also included are those protecting groups specifically adapted for nucleoside and nucleotide chemistry described in Current Protocols in Nucleic Acid Chemistry 06 / 2012, edited by Serge L. Beaucage et al., Chapter 2 of which is incorporated herein by reference in its entirety. Suitable amino protecting groups include, but are not limited to, those described herein and / or in: WO 2018 / 022473, WO 2018 / 098264, WO 2018 / 223056, WO 2018 / 223073, WO 2018 / 223081, WO 2018 / 237194, WO 2019 / 032607, WO 2019 / 055951, and / or WO 2019 / 075357, or U.S. Provisional Patent Application Nos. 62 / 825766 and 62 / 911339 (the description of protecting groups in each of these documents is independently incorporated herein by reference).

[0117] Sample: As used herein, the term "sample" typically refers to an aliquot of material obtained from or derived from a source of interest. In some embodiments, the source of interest is a biological or environmental source. In some embodiments, the source of interest may be or include a cell or organism, such as a microorganism, a plant, or an animal (e.g., a human). In some embodiments, the source of interest is or includes a biological tissue or biological fluid. In some embodiments, the biological tissue or fluid may be or include amniotic fluid, aqueous humor, peritoneal fluid, bile, bone marrow, blood, breast milk, cerebrospinal fluid, earwax, chyle, chyme, ejaculate, endolymph, exudate, feces, gastric acid, gastric juice, lymph, mucus, pericardial fluid, perilymph, peritoneal fluid, pleural fluid, pus, catarrhal secretions, saliva, sebum, semen, serum, smegma, sputum, synovial fluid, sweat, tears, urine, vaginal secretions, vitreous humor, vomit, and / or combinations or components thereof. In some embodiments, the biological fluid may be or include intracellular fluid, extracellular fluid, intravascular fluid (plasma), interstitial fluid, lymph, and / or transcellular fluid. In some embodiments, the biological fluid may be or include plant exudates. In some embodiments, the biological tissue or biological sample may be obtained, for example, by aspiration, biopsy (e.g., fine needle or tissue biopsy), swab (e.g., oral, nasal, skin, or vaginal swab), scraping, surgery, washing, or lavage (e.g., bronchoalveolar, ductal, nasal, ocular, oral, uterine, vaginal, or other washing or lavage). In some embodiments, the biological sample is or comprises cells obtained from an individual. In some embodiments, the sample is a "primary sample" obtained directly from the source of interest by any suitable means. In some embodiments, as will be clear from the context, the term "sample" refers to a preparation obtained by processing the primary sample (e.g., by removing one or more components thereof and / or adding one or more agents thereto), for example, filtration using a semipermeable membrane. Such a "processed sample" may include, for example, nucleic acids or proteins extracted from a sample or obtained by subjecting a primary sample to one or more techniques, such as nucleic acid amplification or reverse transcription, isolation and / or purification of specific components, etc.

[0118] Subject: As used herein, the term "subject" or "test subject" refers to any organism to which a provided compound (e.g., a provided oligonucleotide) or composition is administered in accordance with the present disclosure, e.g., for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans; insects; parasites, etc.) and plants. In some embodiments, the subject is a human. In some embodiments, the subject may be suffering from and / or susceptible to a disease, disorder, and / or condition.

[0119] Substantially: As used herein, the term "substantially" refers to the qualitative condition of exhibiting the entire or nearly entire extent or degree of a desired property or characteristic. A base sequence that is substantially complementary to a second sequence is not identical to the second sequence, but is roughly or nearly identical to the second sequence. Additionally, those skilled in the art of biology and / or chemistry will understand that biological and chemical phenomena rarely, if ever, proceed to completion and / or perfection or achieve or avoid absolute results. Thus, as used herein, the term "substantially" is used to capture the potential lack of completeness inherent in many biological and / or chemical phenomena.

[0120] Sugar: The term "sugar" refers to closed and / or open monosaccharides or polysaccharides. In some embodiments, a sugar is a monosaccharide. In some embodiments, a sugar is a polysaccharide. Sugars include, but are not limited to, ribose, deoxyribose, pentofuranose, pentopyranose, and hexopyranose moieties. As used herein, the term "sugar" also encompasses structural analogs used in place of traditional sugar molecules, such as glycols, polymers that form the backbone of nucleic acid analogs, glycol nucleic acids ("GNAs"). As used herein, the term "sugar" also encompasses structural analogs used in place of natural or naturally occurring nucleotides, such as modified sugars and nucleotide sugars. In some embodiments, a sugar is an RNA or DNA sugar (ribose or deoxyribose). In some embodiments, a sugar is a modified ribose or deoxyribose sugar (e.g., 2'-modified, 5'-modified, etc.). As described herein, in some embodiments, modified sugars can provide one or more desired properties, activities, etc. when used in oligonucleotides and / or nucleic acids. In some embodiments, the sugar is an optionally substituted ribose or deoxyribose. In some embodiments, "sugar" refers to a sugar unit in an oligonucleotide or nucleic acid.

[0121] Susceptible: An individual who is "susceptible" to a disease, disorder, and / or condition is an individual who is at a higher risk than the general population of individuals of developing the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition has a predisposition to having the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may not have been diagnosed with the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may exhibit symptoms of the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may not exhibit symptoms of the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition will develop the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition will not develop the disease, disorder, and / or condition.

[0122] Therapeutic Agent: As used herein, the term "therapeutic agent" generally refers to any agent that induces a desired effect (e.g., a desired biological, clinical, or pharmacological effect) when administered to a subject. In some embodiments, an agent is considered to be a therapeutic agent if it demonstrates a statistically significant effect across a relevant population. In some embodiments, the relevant population is a population of subjects suffering from and / or susceptible to a disease, disorder, or condition. In some embodiments, the relevant population is a population of model organisms. In some embodiments, the relevant population may be defined by one or more criteria, such as age group, sex, genetic background, pre-existing clinical conditions, etc., prior to receiving therapy. In some embodiments, a therapeutic agent is a substance that, when administered to a subject in an effective amount, relieves, ameliorates, alleviates, inhibits, prevents, delays the onset of, reduces the severity of, and / or reduces the incidence of one or more symptoms or characteristics of the disease, disorder, and / or condition in question. In some embodiments, a "therapeutic agent" is a drug that has been approved, or is required to be approved, by a government agency before it can be marketed for administration to humans. In some embodiments, a "therapeutic agent" is a drug for which a prescription is required for administration to a human. In some embodiments, a therapeutic agent is a provided compound, such as a provided oligonucleotide.

[0123] Therapeutically effective amount: As used herein, the term "therapeutically effective amount" refers to an amount of a substance (e.g., a therapeutic agent, composition, and / or formulation) that elicits a desired biological response when administered as part of a treatment regimen. In some embodiments, a therapeutically effective amount of a substance is an amount that, when administered to a subject suffering from or susceptible to a disease, disorder, and / or condition, is sufficient to treat, diagnose, prevent, and / or delay the onset of the disease, disorder, and / or condition. As will be understood by one of skill in the art, the effective amount of a substance can vary depending on factors such as the desired biological endpoint, the substance being delivered, the target cell or tissue, and the like. For example, an effective amount of a compound in a formulation for treating a disease, disorder, and / or condition is an amount that relieves, ameliorate, alleviates, inhibits, prevents, delays the onset of, reduces the severity of, and / or reduces the incidence of one or more symptoms or characteristics of the disease, disorder, and / or condition. In some embodiments, a therapeutically effective amount is administered in a single dose; in some embodiments, multiple unit doses are required to deliver a therapeutically effective amount.

[0124] Treat: As used herein, the terms "treat," "treatment," or "treating" refer to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or characteristics of a disease, disorder, and / or condition. Treatment may be administered to subjects who do not show signs of the disease, disorder, and / or condition. In some embodiments, treatment may be administered to subjects who only show early signs of the disease, disorder, and / or condition, e.g., to reduce the risk of developing pathologies associated with the disease, disorder, and / or condition.

[0125] Unsaturated: The term "unsaturated," as used herein, means that a moiety has one or more units of unsaturation.

[0126] Wild-type: As used herein, the term "wild-type" has its art-recognized meaning and refers to an entity that has structure and / or activity as found in nature in a "normal" (as opposed to mutant, diseased, altered, etc.) state or situation. Those of skill in the art will understand that wild-type genes and polypeptides often exist in multiple alternative forms (e.g., alleles).

[0127] As will be understood by those of skill in the art, the methods and compositions described herein relating to provided compounds (e.g., oligonucleotides) also generally apply to pharmaceutically acceptable salts of such compounds.

[0128] Description of Specific Embodiments Double-stranded oligonucleotides are useful tools for a wide variety of applications. For example, ds oligonucleotides targeting HSD17B13 (e.g., NCBI gene ID: 345275 for human HSD17B13 and related sequences from other organisms) are useful in therapeutic, diagnostic, and research applications, including the treatment of various HSD17B13-related conditions, disorders, and diseases, including, but not limited to, NAFLD, NASH, ASH, alcoholic liver disease, non-alcoholic liver disease, alcoholic cirrhosis, non-alcoholic cirrhosis, steatohepatitis, hepatic steatosis, hepatocellular carcinoma, HCV hepatitis, chronic hepatitis, hereditary hemochromatosis, primary sclerosing cholangitis, drug-induced liver injury, or hepatocellular necrosis. The use of naturally occurring nucleic acids (e.g., unmodified DNA or RNA) is limited, for example, by their susceptibility to endonucleases and exonucleases. Therefore, various synthetic counterparts have been developed to circumvent these drawbacks and / or further improve various properties and activities. These include, inter alia, synthetic oligonucleotides containing chemical modifications, such as base modifications, sugar modifications, backbone modifications, etc., that reduce the susceptibility of these molecules to degradation and improve other properties and / or activities. From a structural standpoint, modifications to the internucleotide linkages can introduce chirality and / or alter charge, and certain properties can be influenced by the positioning of the oligonucleotide's phosphorus linkage atoms. For example, binding affinity, sequence-specific binding to complementary RNA, stability against nucleases, target nucleic acid cleavage, delivery, pharmacokinetics, etc., can be influenced, inter alia, by the chirality and / or charge of the backbone linkage atoms.

[0129] In some embodiments, the ds oligonucleotide targeting HSD17B13 is (1) a guide strand containing a backbone phosphorothioate chiral center in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the (N-2) nucleotide immediately upstream, i.e., in the 5' direction; (2) a guide strand containing Rp, Sp, or alternating backbone phosphorothioate chiral centers between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide in the 3' direction, and between the +2 nucleotide and the immediately downstream (+3) nucleotide; (3) a guide strand that contains one or more backbone phosphorothioate chiral centers in the Sp configuration upstream, i.e., in the 5' direction, of a backbone phosphorothioate chiral center between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide, wherein the upstream backbone phosphorothioate chiral center is in the Rp or Sp configuration; (4) a guide strand containing one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide, and between (a) the +3 and +4 nucleotides and (b) the +5 and +6 nucleotides; (5) a passenger strand in combination with one or more of the aforementioned guide strands, comprising one or more backbone chiral centers of Rp or Sp configuration; and (6) a passenger strand, combined with one or more of the aforementioned guide strands, containing a backbone phosphorothioate chiral center of the Sp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide in the 3' direction, and between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide; and ds oligonucleotides are (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and further comprising one or more of: The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide bond. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bond incorporated into the guide strand or passenger strand is an Rp non-negatively charged internucleotide bond. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bond is an Sp non-negatively charged internucleotide bond. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bond is a stereorandom non-negatively charged internucleotide bond.

[0130] In some embodiments, the ds oligonucleotide targeting HSD17B13 comprises: (1) a phosphorothioate chiral center in the Rp or Sp configuration; (2) an Rp, Sp, or stereorandom non-negatively charged internucleotide linkage, wherein the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide linkage comprises a 2' modification, e.g., 2'F; and (3) (a) Including, but not limited to: [ka] 5'PO modifications such as; (b) Without limitation, [ka] 5'VP modifications such as; (c) Without limitation, [ka] 5'MeP modifications such as; (d) Without limitation, [ka] 5'PN and 5'triazole-P modifications such as wherein the bases are selected from A, C, G, T, U, abasic and modified nucleobases; R 2’ is H, OH, O-alkyl, F, MOE, locked nucleic acid (LNA) bridges, and bridged nucleic acid (BNA) bridges to the 4'C, including, but not limited to: [ka] (selected from In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0131] In some embodiments, the ds oligonucleotide targeting HSD17B13 comprises: (1) a phosphorothioate chiral center in the Rp or Sp configuration; (2) an Rp, Sp, or stereorandom non-negatively charged internucleotide linkage, wherein the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide linkage comprises a 2' modification, e.g., 2'F; and (3) (a) Including, but not limited to: [ka] 5'PO nucleotides such as; (b) Without limitation, [ka] 5'VP nucleotides such as; (c) Without limitation, [ka] 5'MeP nucleotides such as; (d) Without limitation, [ka] 5'PN and 5'triazole-P nucleotides such as; (e) Without limitation, [ka] 5' abasic VP and 5' abasic MeP nucleotides such as In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0132] In some embodiments, the ds oligonucleotide targeting HSD17B13 is (1) a guide strand containing a backbone phosphorothioate chiral center in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the (N-2) nucleotide immediately upstream, i.e., in the 5' direction; (2) a guide strand containing Rp, Sp, or alternating backbone phosphorothioate chiral centers between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide in the 3' direction, and between the +2 nucleotide and the immediately downstream (+3) nucleotide; (3) a guide strand that contains one or more backbone phosphorothioate chiral centers in the Sp configuration upstream, i.e., in the 5' direction, of a backbone phosphorothioate chiral center between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide, wherein the upstream backbone phosphorothioate chiral center is in the Rp or Sp configuration; (4) a guide strand containing one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide, and between (a) the +3 and +4 nucleotides and (b) the +5 and +6 nucleotides; (5) a passenger strand in combination with one or more of the aforementioned guide strands, comprising one or more backbone chiral centers of Rp or Sp configuration; and (6) a passenger strand, combined with one or more of the aforementioned guide strands, containing a backbone phosphorothioate chiral center of the Sp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide in the 3' direction, and between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide; and ds oligonucleotides are (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and further comprising one or more of: The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide bond. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bond incorporated into the guide strand or passenger strand is an Rp non-negatively charged internucleotide bond. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bond is an Sp non-negatively charged internucleotide bond. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bond is a stereorandom non-negatively charged internucleotide bond.

[0133] In some embodiments, the ds oligonucleotide targeting HSD17B13 contains a non-natural internucleotide bond, such as a neutral internucleotide bond, which, in certain embodiments, can be used to attach one or more molecules to the double-stranded oligonucleotide described herein. In certain embodiments, such a binding molecule can facilitate targeting and / or delivery of the double-stranded oligonucleotide. For example, but not limited to, such a binding molecule includes a lipophilic molecule. In certain embodiments, the binding molecule is a molecule containing one or more GalNAc moieties. In certain embodiments, the binding molecule is a receptor. In certain embodiments, the binding molecule is a receptor ligand.

[0134] In certain embodiments, the present disclosure provides techniques (e.g., compounds, methods, etc.) for improving the stability of oligonucleotides while maintaining or increasing activity, including compositions of oligonucleotides with improved stability.

[0135] In certain embodiments, the present disclosure provides techniques for incorporating various additional chemical moieties into ds oligonucleotides. In certain embodiments, the present disclosure provides, for example, reagents and methods for introducing additional chemical moieties with a nucleobase (e.g., by covalent attachment to a site on the nucleobase, optionally via a linker).

[0136] In certain embodiments, the present disclosure provides techniques, e.g., ds oligonucleotide compositions and methods, for achieving allele-specific suppression, in which transcripts from one allele of a particular target gene are selectively knocked down relative to at least one other allele of the same gene.

[0137] In particular, the present disclosure provides structural elements, techniques, and / or features that can be incorporated into ds oligonucleotides and that can confer or adjust one or more properties thereof (e.g., compared to an otherwise identical ds oligonucleotide lacking the related technique or feature). In certain embodiments, the present disclosure describes that one or more of the provided techniques and / or features can be usefully incorporated into ds oligonucleotides of various sequences.

[0138] In certain embodiments, the present disclosure demonstrates that certain provided structural elements, techniques, and / or features are particularly useful for ds oligonucleotides (e.g., RNAi agents) that participate in and / or induce the RNAi machinery. Nevertheless, the teachings of the present disclosure are not limited to ds oligonucleotides that participate in or act via any particular mechanism.

[0139] In certain embodiments, the present disclosure relates to any ds oligonucleotide useful for any purpose, acting via any mechanism, and comprising any sequence, structure, or format (or portion thereof) described herein.

[0140] In some embodiments, the ds oligonucleotide targeting HSD17B13 contains backbone phosphorothioate chiral centers in the Sp configuration between the 3' terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide; (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide linkage, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, where n is about 1 to 49. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0141] In some embodiments, the ds oligonucleotides targeting HSD17B13 contain Rp, Sp or alternating backbone phosphorothioate chiral centers between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide, and (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide linkage, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, where n is about 1 to 49. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0142] In some embodiments, the present disclosure provides one or more backbone phosphorothioate chiral centers of Rp or Sp configuration upstream of a backbone phosphorothioate chiral center of Sp configuration between the 3' terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide; and (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and providing a ds oligonucleotide targeting HSD17B13, comprising one or more of: wherein the ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide linkage, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages (where n is about 1 to 49). In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0143] In some embodiments, the guide strand comprises one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds between the second (+2) nucleotide and the third (+3) nucleotide from the 5'-terminal nucleotide of the guide strand, and an internucleotide bond to the 3'(N-1) nucleotide immediately preceding the terminal end; (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide linkage, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, where n is about 1 to 49. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0144] In some embodiments, the guide strand contains backbone phosphorothioate chiral centers in the Sp configuration between the 3' terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide; (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide bond, and the passenger strand comprises one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0145] In some embodiments, the guide strand contains Rp, Sp, or alternating backbone phosphorothioate chiral centers between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide; (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by an Rp, Sp, or stereorandom non-negatively charged internucleotide bond, and the passenger strand comprises one or more backbone chiral centers in the Rp or Sp configuration. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages. In certain embodiments disclosed herein, the backbone phosphorothioate chiral centers are in the Rp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide. In certain embodiments disclosed herein, the backbone phosphorothioate chiral centers are in the Sp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide. In certain embodiments, the backbone phosphorothioate chiral centers are in the Rp and Sp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide, respectively. In certain embodiments disclosed herein, the backbone phosphorothioate chiral centers are in the Sp, Rp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide, respectively.

[0146] In some embodiments, the guide strand comprises one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration upstream of a backbone phosphorothioate chiral center in the Sp configuration between the 3' terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide; (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by an Rp, Sp, or stereorandom non-negatively charged internucleotide bond, and the passenger strand comprises one or more backbone chiral centers in an Rp or Sp configuration. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0147] In some embodiments, the guide strand comprises one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the (+2) nucleotide and the immediately downstream (+3) nucleotide, and between (a) the (+3) nucleotide and the (+4) nucleotide and (b) the (+5) nucleotide and the (+6) nucleotide; (1) a guide strand in which one or both of the 5'- and 3'-terminal dinucleotides are not linked by a non-negatively charged internucleotide bond, i.e., the guide strand includes one or more non-negatively charged internucleotide bond(s) downstream of the linkage between the 5'-terminal dinucleotides, i.e., in the 3' direction, and / or upstream of the linkage between the 3'-terminal dinucleotides, i.e., in the 5' direction; (2) A guide strand in which one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds exist between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide); (3) a guide strand in which an Rp, Sp, or stereorandom non-negatively charged internucleotide bond is present between the third (+3) nucleotide and the fourth (+4) nucleotide from the 5'-terminal nucleotide of the guide strand, and / or between the tenth (+10) nucleotide and the eleventh (+11) nucleotide from the 5'-terminal nucleotide of the guide strand; (4) a passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds upstream of the central nucleotide of the passenger strand, i.e., in the 5' direction; and (5) A passenger strand having one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds downstream of the central nucleotide of the passenger strand, i.e., in the 3' direction. and The ds oligonucleotide further comprises a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by an Rp, Sp, or stereorandom non-negatively charged internucleotide bond, and the passenger strand comprises one or more backbone chiral centers in an Rp or Sp configuration. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0148] In some embodiments, the guide strand comprises one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3'(N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide), a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide linkage, and the passenger strand comprises one or more backbone chiral centers in the Rp or Sp configuration.

[0149] In certain embodiments, the guide strand comprises backbone phosphorothioate chiral centers in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide, and 2' modifications, e.g., 2'F modifications, of the 3' nucleotides of nucleotide pairs linked by Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages (where n is about 1 to 49) and one or more backbone chiral centers in the Rp or Sp configuration.

[0150] In certain embodiments, the guide strand comprises Rp, Sp, or alternating backbone phosphorothioate chiral centers between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the (+2) nucleotide and the immediately downstream (+3) nucleotide, and 2' modifications, e.g., 2'F modifications, of the 3' nucleotides of nucleotide pairs linked by Rp, Sp, or stereorandom non-negatively charged internucleotide linkages; and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages (where n is about 1 to 49) and one or more backbone chiral centers in the Rp or Sp configuration. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more of Rp, Sp or stereorandom non-negatively charged internucleotide linkages is a stereorandom non-negatively charged internucleotide linkage.

[0151] In certain embodiments, the guide strand comprises one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration upstream of the backbone phosphorothioate chiral center in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide, a 2' modification, e.g., a 2'F modification, of the 3' nucleotide of the nucleotide pair linked by the Rp, Sp, or stereorandom non-negatively charged internucleotide linkage, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages (where n is about 1 to 49) and one or more backbone chiral centers in the Rp or Sp configuration. In certain embodiments, the one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand or passenger strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are stereorandom non-negatively charged internucleotide linkages.

[0152] In certain embodiments, the guide strand comprises one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, or 2' modifications, e.g., 2'F modifications, of the 3' nucleotide of a nucleotide pair linked by an Rp, Sp, or stereorandom non-negatively charged internucleotide linkage, present between any two adjacent nucleotides between the second (+2) nucleotide from the 5'-terminal nucleotide of the guide strand and the 3'(N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide), and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages (where n is about 1 to 49) and one or more backbone chiral centers of an Rp or Sp configuration. In certain embodiments, the one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages incorporated into the guide strand are Rp non-negatively charged internucleotide linkages. In certain embodiments, the one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages are Sp non-negatively charged internucleotide linkages. In certain embodiments, one or more of Rp, Sp or stereorandom non-negatively charged internucleotide linkages is a stereorandom non-negatively charged internucleotide linkage.

[0153] In certain embodiments, the internucleotide linkages of the oligonucleotide comprise or consist of 1 to 5, 1 to 10, 1 to 15, 1 to 20, 1 to 25, 1 to 30, 1 to 40, 1 to 50, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more chiral-controlled internucleotide linkages. In certain embodiments, the present disclosure provides dsRNAi oligonucleotide compositions in which the dsRNAi oligonucleotide comprises at least one chiral-controlled internucleotide linkage. In certain embodiments, the present disclosure provides dsRNAi oligonucleotide compositions in which the dsRNAi oligonucleotide is stereorandom or not chirally controlled. In certain embodiments, at least one internucleotide linkage in the dsRNAi oligonucleotide is stereorandom and at least one internucleotide linkage is chirally controlled.

[0154] In certain embodiments, the internucleotide linkages of the oligonucleotide comprise or consist of one or more neutrally charged internucleotide linkages.

[0155] HSD17B13 In some embodiments, HSD17B13 refers to a gene or its gene product (including, but not limited to, nucleic acids, including but not limited to, DNA or RNA, transcripts, and proteins encoded thereby; may be from any form of HSD17B13, e.g., wild-type or mutant alleles) from any species, and may be known as ****. In some embodiments, it refers to the gene and its product in humans. In some embodiments, it refers to the gene and its product in non-human primates. Various HSD17B13 sequences, including variants thereof, from humans, mice, rats, monkeys, etc., are readily available to those skilled in the art. In some embodiments, HSD17B13 is human or mouse HSD17B13, which may be wild-type or mutant. It has been reported that HSD17B13 can have many functions. Various techniques, such as assays, cells, animal models, etc., have also been reported and can be used to characterize and / or evaluate the provided techniques (e.g., oligonucleotides, compositions, methods, etc.) of the present disclosure.

[0156] In some embodiments, a variant or isoform of the HSD17B13 gene, transcript (e.g., pre- or post-spliced ​​mRNA) or protein comprises a mutation. In some embodiments, the HSD17B13 gene, transcript or protein is an alternatively spliced ​​variant or isoform or a product of its transcription or translation.

[0157] HSD17B13-associated pathological conditions, disorders, or diseases Various conditions, disorders, or diseases have been reported to be associated with HSD17B13. Generally, a disease, disorder, or condition is associated with HSD17B13 if the presence, level, activity, and / or form of HSD17B13 and / or its products (e.g., transcripts, encoded proteins, etc.) correlates with the incidence of and / or susceptibility to that disease, disorder, or condition (e.g., across a relevant population). In some embodiments, HSD17B13-associated conditions, disorders, or diseases can be treated and / or prevented by reducing the expression, level, and / or activity of HSD17B13 transcripts and / or proteins.

[0158] Various HSD17B13-associated pathologies, disorders or diseases have been reported. In some embodiments, the HSD17B13-associated pathologies, disorders or diseases are NAFLD, NASH, ASH, alcoholic liver disease, non-alcoholic liver disease, alcoholic cirrhosis, non-alcoholic cirrhosis, steatohepatitis, hepatic steatosis, hepatocellular carcinoma, HCV hepatitis, chronic hepatitis, hereditary hemochromatosis, primary sclerosing cholangitis, drug-induced liver injury or hepatocellular necrosis. In some embodiments, the HSD17B13-associated pathologies, disorders or diseases are NAFLD. In some embodiments, the HSD17B13-associated pathologies, disorders or diseases are NASH.

[0159] In particular, the provided technology is useful for treating or preventing HSD17B13-associated conditions, disorders, or diseases, such as NAFLD, NASH, ASH, alcoholic liver disease, non-alcoholic liver disease, alcoholic cirrhosis, non-alcoholic cirrhosis, steatohepatitis, hepatic steatosis, hepatocellular carcinoma, HCV hepatitis, chronic hepatitis, hereditary hemochromatosis, primary sclerosing cholangitis, drug-induced liver injury, or hepatocellular necrosis, etc. In some embodiments, the present disclosure relates to the use of a ds oligonucleotide targeting HSD17B13 or a composition thereof in the treatment of HSD17B13-associated disorders, diseases, or conditions, such as NAFLD, NASH, ASH, alcoholic liver disease, non-alcoholic liver disease, alcoholic cirrhosis, non-alcoholic cirrhosis, steatohepatitis, hepatic steatosis, hepatocellular carcinoma, HCV hepatitis, chronic hepatitis, hereditary hemochromatosis, primary sclerosing cholangitis, drug-induced liver injury, or hepatocellular necrosis, etc.

[0160] In some embodiments, treatment or prevention with the provided technologies reduces the rate of HSD17B13 production, reduces, stops, or reverses the accumulation of HSD17B13, hi some embodiments, treatment or prevention with the provided technologies reduces the rate of clinical deterioration, or delays or prevents the onset of a condition, disorder, or disease.

[0161] As will be appreciated by those skilled in the art, the present disclosure provides that the characterization / assessment of such pathologies, disorder or disease mechanisms, genotypes, symptoms, biomarkers, etc. may be utilized.

[0162] double-stranded oligonucleotides In particular, the present disclosure provides oligonucleotides of various designs, which may include various nucleobases and their patterns, sugars and their patterns, internucleotide linkages and their patterns, and / or additional chemical moieties and their patterns, as described herein. In some embodiments, the provided ds oligonucleotides targeting HSD17B13 can induce a decrease in the expression, level, and / or activity of the HSD17B13 gene and / or one or more of its products (e.g., transcripts, mRNA, proteins, etc.). In some embodiments, the provided ds oligonucleotides targeting HSD17B13 can induce a decrease in the expression, level, and / or activity of the HSD17B13 gene and / or one or more of its products in cells of a subject or patient. In some embodiments, the cells normally express HSD17B13 or produce HSD17B13 protein. In some embodiments, the provided ds oligonucleotides targeting HSD17B13 are capable of inducing a decrease in the expression, level and / or activity of an HSD17B13 target gene or gene product, and have a base sequence consisting of, comprising, or comprising a portion thereof (e.g., a span of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more contiguous bases) of the base sequence of a ds oligonucleotide targeting HSD17B13 disclosed herein, wherein each T can be independently replaced with U, and vice versa, and the ds oligonucleotide comprises at least one non-naturally occurring modification of the base, sugar and / or internucleotide linkage.

[0163] In some embodiments, ds oligonucleotides targeting HSD17B13 can induce a decrease in the expression, level, and / or activity of a target gene, such as an HSD17B13 target gene, or its product. In some embodiments, ds oligonucleotides targeting HSD17B13 can induce a decrease in the expression, level, and / or activity of an HSD17B13 target gene or its product by RNase H-mediated knockdown. In some embodiments, ds oligonucleotides targeting HSD17B13 can induce a decrease in the expression, level, and / or activity of an HSD17B13 target gene or its product by sterically blocking translation after binding to the HSD17B13 target gene mRNA and / or by altering or interfering with mRNA splicing. Nevertheless, the present disclosure is not limited to any particular mechanism. In some embodiments, the present disclosure provides oligonucleotides, compositions, methods, etc. that can be manipulated by double-stranded RNA interference.

[0164] In some embodiments, ds oligonucleotides targeting HSD17B13 can mediate a decrease in the expression, level and / or activity of HSD17B13. In some embodiments, ds oligonucleotides targeting HSD17B13 can mediate a decrease in the expression, level and / or activity of HSD17B13 protein. In some embodiments, ds oligonucleotides targeting HSD17B13 can mediate a decrease in the level of HSD17B13 protein.

[0165] In some embodiments, ds oligonucleotides targeting HSD17B13 can mediate a decrease in HSD17B13 expression, levels and / or activity by a mechanism involving degradation of the mRNA.

[0166] In some embodiments, ds oligonucleotides targeting HSD17B13 are capable of mediating a decrease in the expression, level and / or activity of two or more HSD17B13 alleles.

[0167] In some embodiments, the present disclosure relates to a method of treating an HSD17B13-associated disease, disorder, or condition in which HSD17B13 is overexpressed, comprising administering a therapeutically effective amount of a ds oligonucleotide targeting HSD17B13, which can mediate a decrease in the expression, level, and / or activity of HSD17B13. In some embodiments, multiple forms, e.g., alleles, of HSD17B13 may exist, and the provided techniques can reduce the expression, level, and / or activity of two or more or all of such forms and their products.

[0168] In some embodiments, the present disclosure relates to a method of treating a disease, disorder or condition associated with HSD17B13, comprising administering a therapeutic amount of a ds oligonucleotide targeting HSD17B13 that is capable of mediating a decrease in expression, level and / or activity of HSD17B13.

[0169] In some embodiments, ds oligonucleotides targeting HSD17B13 can mediate a decrease in expression, levels and / or activity of HSD17B13 by a mechanism involving splicing modulation, eg, exon skipping.

[0170] In some embodiments, ds oligonucleotides targeting HSD17B13 comprise structural elements or portions thereof described herein, e.g., in Table 1. In some embodiments, ds oligonucleotides targeting HSD17B13 comprise a base sequence (or portion thereof) described herein (wherein each T may be independently replaced with U, and vice versa), a chemical modification or chemical modification pattern (or portion thereof), and / or a format or portion thereof described herein. In some embodiments, ds oligonucleotides targeting HSD17B13 have a base sequence comprising a base sequence (or portion thereof) (wherein each T may be independently replaced with U), a chemical modification pattern (or portion thereof), and / or a format of an oligonucleotide disclosed herein, e.g., in Table 1, or otherwise disclosed herein. In some embodiments, such oligonucleotides, e.g., ds oligonucleotides targeting HSD17B13, reduce the expression, level, and / or activity of a gene, e.g., the HSD17B13 gene, or its gene product.

[0171] In particular, ds oligonucleotides targeting HSD17B13 can hybridize to their target nucleic acids (e.g., pre-mRNA, mature mRNA, etc.). For example, in some embodiments, ds oligonucleotides targeting HSD17B13 can hybridize to HSD17B13 nucleic acids derived from DNA strands (either strand of the HSD17B13 gene). In some embodiments, ds oligonucleotides targeting HSD17B13 can hybridize to HSD17B13 transcripts. In some embodiments, ds oligonucleotides targeting HSD17B13 can hybridize to HSD17B13 nucleic acids at any stage of RNA processing, including, but not limited to, pre-mRNA or mature mRNA. In some embodiments, ds oligonucleotides targeting HSD17B13 can hybridize to any element of an HSD17B13 nucleic acid or its complement, including, but not limited to, a promoter region, an enhancer region, a transcription termination region, a translation initiation signal, a translation termination signal, a coding region, a non-coding region, an exon, an intron, an intron / exon or an exon / intron junction, a 5'UTR, or a 3'UTR. In some embodiments, ds oligonucleotides targeting HSD17B13 can hybridize to their targets with no more than two mismatches. In some embodiments, ds oligonucleotides targeting HSD17B13 can hybridize to their targets with no more than one mismatch. In some embodiments, ds oligonucleotides targeting HSD17B13 can hybridize to their targets with no mismatches (e.g., all CG and / or AT / U base pairings).

[0172] In some embodiments, oligonucleotide can hybridize to two or more variants of transcript.In some embodiments, ds oligonucleotide that targets HSD17B13 can hybridize to two or more or all variants of HSD17B13 transcript.In some embodiments, ds oligonucleotide that targets HSD17B13 can hybridize to two or more or all variants of HSD17B13 transcript from sense strand.

[0173] In some embodiments, the HSD17B13 target of the HSD17B13-targeting ds oligonucleotide is HSD17B13 RNA, which is not mRNA.

[0174] In some embodiments, oligonucleotides, e.g., ds oligonucleotides targeting HSD17B13, contain increased levels of one or more isotopes. In some embodiments, oligonucleotides, e.g., ds oligonucleotides targeting HSD17B13, are labeled, for example, with one or more isotopes of one or more elements (e.g., hydrogen, carbon, nitrogen, etc.). In some embodiments, oligonucleotides in the provided compositions, e.g., ds oligonucleotides targeting HSD17B13 (e.g., multiple oligonucleotides in the composition), comprise base modifications, sugar modifications, and / or internucleotide linkage modifications, and the oligonucleotides contain enriched levels of deuterium. In some embodiments, oligonucleotides, e.g., ds oligonucleotides targeting HSD17B13, are labeled with deuterium at one or more positions (- 1 H is- 2 In some embodiments, one or more of the oligonucleotide strands or any moieties conjugated to the oligonucleotide strands (e.g., targeting moieties, etc.) 1 H, 2 substituted with H. Such oligonucleotides may be used in the compositions and methods described herein.

[0175] In some embodiments, the present disclosure provides: 1) have a common base sequence complementary to a target sequence in the transcript (e.g., an HSD17B13 target sequence); and 2) containing one or more modified sugar moieties and / or modified internucleotide linkages; An oligonucleotide composition is provided that includes a plurality of oligonucleotides.

[0176] In some embodiments, ds oligonucleotides targeting HSD17B13 that share a common base sequence may have the same pattern of nucleotide modifications, such as sugar modifications, base modifications, etc. In some embodiments, the nucleotide modification pattern may be represented by a combination of position and modification. In some embodiments, the backbone linkage pattern includes the position and type (e.g., phosphate, phosphorothioate, substituted phosphorothioate, etc.) of each internucleotide linkage.

[0177] In some embodiments, for example, in the provided compositions, multiple oligonucleotides are of the same oligonucleotide type. In some embodiments, oligonucleotides of an oligonucleotide type have a common sugar modification pattern. In some embodiments, oligonucleotides of an oligonucleotide type have a common base modification pattern. In some embodiments, oligonucleotides of an oligonucleotide type have a common nucleoside modification pattern. In some embodiments, oligonucleotides of an oligonucleotide type have the same structure. In some embodiments, oligonucleotides of an oligonucleotide type are identical. In some embodiments, multiple oligonucleotides are identical. In some embodiments, multiple oligonucleotides share the same structure.

[0178] In some embodiments, as exemplified herein, the ds oligonucleotide that targets HSD17B13 is chiral controlled and comprises one or more chiral controlled internucleotide bonds.In some embodiments, the ds oligonucleotide that targets HSD17B13 is stereochemically pure.In some embodiments, the ds oligonucleotide that targets HSD17B13 is substantially separated from other stereoisomers.

[0179] In some embodiments, ds oligonucleotides targeting HSD17B13 comprise one or more modified nucleobases, one or more modified sugars, and / or one or more modified internucleotide linkages.

[0180] In some embodiments, the ds oligonucleotide targeting HSD17B13 comprises one or more modified sugars. In some embodiments, the oligonucleotide of the present disclosure comprises one or more modified nucleobases. Various modifications can be introduced into the sugar and / or nucleobase according to the present disclosure. For example, in some embodiments, the modification is the modification described in U.S. Patent No. 9,006,198. In some embodiments, the modification is as described in U.S. Patent No. 9,394,333, U.S. Patent No. 9,744,183, U.S. Patent No. 9,605,019, U.S. Patent No. 9,598,458, U.S. Patent No. 9,982,257, U.S. Patent No. 10,160,969, U.S. Patent No. 10,479,995, U.S. Patent Application Publication No. 2020 / 0056173, U.S. Patent Application Publication No. 2018 / 0216107, U.S. Patent Application Publication No. 2019 / 0127733, U.S. Patent Application Publication No. 10,450,568, U.S. Patent Application Publication No. 2019 / 0077817, U.S. Patent Application Publication No. 2019 / 0249173, U.S. Patent Application Publication No. 2019 / 037 5774, WO 2018 / 223056, WO 2018 / 223073, WO 2018 / 223081, WO 2018 / 237194, WO 2019 / 032607, WO 2019 / 055951, WO 2019 / 075357, WO 2019 / 200185, WO 2019 / 217784, WO 2019 / 032612, and / or WO 2020 / 191252, each sugar, base, and internucleotide linkage modification being independently incorporated herein by reference.

[0181] As used in this disclosure, in some embodiments, "one or more" is 1 to 200, 1 to 150, 1 to 100, 1 to 90, 1 to 80, 1 to 70, 1 to 60, 1 to 50, 1 to 40, 1 to 30, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25. In some embodiments, "one or more" is 1. In some embodiments, "one or more" is 2. In some embodiments, "one or more" is 3. In some embodiments, "one or more" is 4. In some embodiments, "one or more" is 5. In some embodiments, "one or more" is 6. In some embodiments, "one or more" is 7. In some embodiments, "one or more" is 8. In some embodiments, "one or more" is 9. In some embodiments, "one or more" is 10. In some embodiments, "one or more" is at least 1. In some embodiments, "one or more" is at least 2. In some embodiments, "one or more" is at least 3. In some embodiments, "one or more" is at least 4. In some embodiments, "one or more" is at least 5. In some embodiments, "one or more" is at least 6. In some embodiments, "one or more" is at least 7. In some embodiments, "one or more" is at least 8. In some embodiments, "one or more" is at least 9. In some embodiments, "one or more" is at least 10.

[0182] As used in this disclosure, in some embodiments, "at least one" is 1 to 200, 1 to 150, 1 to 100, 1 to 90, 1 to 80, 1 to 70, 1 to 60, 1 to 50, 1 to 40, 1 to 30, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25. In some embodiments, "at least one" is 1. In some embodiments, "at least one" is 2. In some embodiments, "at least one" is 3. In some embodiments, "at least one" is 4. In some embodiments, "at least one" is 5. In some embodiments, "at least one" is 6. In some embodiments, "at least one" is 7. In some embodiments, "at least one" is 8. In some embodiments, "at least one" is 9. In some embodiments, "at least one" is ten.

[0183] In some embodiments, the HSD17B13-targeted ds oligonucleotide is or comprises a HSD17B13-targeted ds oligonucleotide listed in Table 1.

[0184] As demonstrated in the present disclosure, in some embodiments, the provided oligonucleotide (e.g., a ds oligonucleotide targeting HSD17B13) is characterized in that it knocks down its target (e.g., an HSD17B13 transcript for a ds oligonucleotide targeting HSD17B13) when contacted with the transcript in a knockdown system.

[0185] In some embodiments, the ds oligonucleotide is provided in the form of a salt. In some embodiments, the ds oligonucleotide is provided as a salt containing a negatively charged internucleotide linkage (e.g., a phosphorothioate internucleotide linkage, a natural phosphate linkage, etc.) that exists in salt form. In some embodiments, the ds oligonucleotide is provided as a pharmaceutically acceptable salt. In some embodiments, the ds oligonucleotide is provided as a metal salt. In some embodiments, the oligonucleotide is provided as a sodium salt. In some embodiments, the ds oligonucleotide is provided as a metal salt (e.g., a sodium salt), and each negatively charged internucleotide linkage independently exists in salt form (e.g., in the case of a sodium salt, -OP(O)(SNa)-O- for a phosphorothioate internucleotide linkage, -OP(O)(ONa)-O- for a natural phosphate linkage, etc.).

[0186] Double-stranded oligonucleotide sequence In some embodiments, ds oligonucleotides targeting HSD17B13 comprise a base sequence described herein or a portion thereof (e.g., 5-50, 5-40, 5-30, 5-20, or a span of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 30, or at least 10, at least 15 contiguous nucleobases) with 0-5 (e.g., 0, 1, 2, 3, 4, or 5) mismatches, where each T can be independently replaced with U, or vice versa. In some embodiments, ds oligonucleotides targeting HSD17B13 comprise a base sequence described herein or a portion thereof, where the portion is a span of at least 10 contiguous nucleobases, or a span of at least 15 contiguous nucleobases, with 1-5 mismatches. In some embodiments, a ds oligonucleotide targeting HSD17B13 comprises a base sequence described herein or a portion thereof, wherein the portion is a span of at least 10 contiguous nucleobases, or a span of at least 10 contiguous nucleobases, having 1 to 5 mismatches (wherein each T can be independently replaced with U, or vice versa). In some embodiments, the base sequence of the oligonucleotide comprises or consists of 10 to 50 (e.g., about or at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45; in some embodiments, at least 15; in some embodiments, at least 16; in some embodiments, at least 17; in some embodiments, at least 18; in some embodiments, at least 19; in some embodiments, at least 20; in some embodiments, at least 21; in some embodiments, at least 22; in some embodiments, at least 23; in some embodiments, at least 24; in some embodiments, at least 25) contiguous bases of a base sequence identical to or complementary to a base sequence (e.g., in an intron) of the HSD17B13 gene or a transcript (e.g., mRNA) thereof.

[0187] As will be understood by those skilled in the art, the base sequence of a ds oligonucleotide targeting HSD17B13 is typically of sufficient length and complementarity to its target, e.g., an RNA transcript (e.g., pre-mRNA, mature mRNA, etc.), to mediate target-specific knockdown. In some embodiments, the base sequence of a ds oligonucleotide targeting HSD17B13 is of sufficient length and identity to the HSD17B13 transcript target to mediate target-specific knockdown. In some embodiments, the ds oligonucleotide targeting HSD17B13 is complementary to a portion of the HSD17B13 transcript (HSD17B13 transcript target sequence). In some embodiments, the base sequence of a ds oligonucleotide targeting HSD17B13 has 90% or more identity to the base sequence of an oligonucleotide disclosed in Table 1 (wherein each T can be independently replaced with U, and vice versa). In some embodiments, the base sequence of a ds oligonucleotide targeting HSD17B13 has 95% or greater identity to the base sequence of an oligonucleotide disclosed in Table 1 (wherein each T may be independently replaced with U, or vice versa). In some embodiments, the base sequence of a ds oligonucleotide targeting HSD17B13 comprises a contiguous span of 15 or more bases of an oligonucleotide disclosed in Table 1 (wherein each T may be independently replaced with U, or vice versa), except where one or more bases within the span are abasic (e.g., a nucleobase is absent from the nucleotide). In some embodiments, the base sequence of a ds oligonucleotide targeting HSD17B13 comprises a contiguous span of 19 or more bases of a ds oligonucleotide targeting HSD17B13 disclosed herein, except where one or more bases within the span are abasic (e.g., a nucleobase is absent from the nucleotide). In some embodiments, the base sequence of a ds oligonucleotide targeting HSD17B13 comprises a contiguous span of 19 or more bases of an oligonucleotide disclosed herein, except for one or two base differences at the 5' and / or 3' ends of the base sequence (wherein each T can be independently replaced with a U, and vice versa).

[0188] In some embodiments, the present disclosure relates to oligonucleotides having a base sequence comprising the base sequence of any of the oligonucleotides disclosed herein, where each T can be independently replaced by U, and vice versa.

[0189] In some embodiments, the present disclosure relates to oligonucleotides having a base sequence comprising at least 15 contiguous bases of the base sequence of any of the oligonucleotides disclosed herein, wherein each T can be independently replaced by U, and vice versa.

[0190] In some embodiments, the present disclosure relates to oligonucleotides having a base sequence that is at least 90% identical to the base sequence of any oligonucleotide disclosed herein, where each T can be independently replaced by U, and vice versa.

[0191] In some embodiments, the present disclosure relates to oligonucleotides having a base sequence that is at least 95% identical to the base sequence of any oligonucleotide disclosed herein, where each T can be independently replaced by U, and vice versa.

[0192] In some embodiments, the ds oligonucleotide targeting HSD17B13 is selected from Table 1.

[0193] In some embodiments, the base sequence of the ds oligonucleotide targeted to HSD17B13 is complementary to the HSD17B13 transcript or a portion thereof.

[0194] In some embodiments, the base sequence of a ds oligonucleotide targeting HSD17B13 is complementary to a portion of an HSD17B13 nucleic acid sequence, such as an HSD17B13 gene sequence, an HSD17B13 transcript, or an HSD17B13 mRNA sequence. In some embodiments, a ds oligonucleotide targeting HSD17B13 is identical to a portion of an HSD17B13 nucleic acid sequence, such as an HSD17B13 gene sequence, an HSD17B13 transcript, or an HSD17B13 mRNA sequence. In some embodiments, the base sequence of such a portion is characteristic of HSD17B13 in that no other genomic or transcript sequence in the system contains the same sequence as that portion. In some embodiments, no other genomic or transcript sequence in the system contains a sequence that differs from such a portion by no more than one nucleobase. In some embodiments, no other genomic or transcript sequence in the system contains a sequence that differs from such a portion by no more than two nucleobases. In some embodiments, the portion of the gene to which the oligonucleotide is complementary is referred to as the target sequence of the oligonucleotide. In some embodiments, the system is or includes a cell, sample, tissue, organ, or species. For example, for an oligonucleotide targeting human HSD17B13, the relevant species in many embodiments is human. In some embodiments, the system can be or includes multiple species, for example, when cross-species activity and / or properties are characterized and / or evaluated. In some embodiments, the portion is in an exon. In some embodiments, the portion is in an intron. In some embodiments, the portion spans an intron and an exon. In some embodiments, the portion spans two exons. In some embodiments, the portion is in the 5'-UTR region. In some embodiments, the portion is in the 3'-UTR region.

[0195] In some embodiments, the ds oligonucleotides targeting HSD17B13 target two or more or all alleles of HSD17B13 (if multiple alleles are present in the relevant system), hi some embodiments, the oligonucleotides reduce the expression, levels and / or activity of both wild-type and mutant HSD17B13 and / or their transcripts and / or products.

[0196] In some embodiments, the base sequences of the provided oligonucleotides are perfectly complementary to HSD17B13 target sequences in both humans and non-human primates (NHPs). In some embodiments, such sequences may be particularly useful because they can be easily evaluated in both humans and non-human primates.

[0197] In some embodiments, ds oligonucleotides targeting HSD17B13 comprise a base sequence or portion thereof set forth in the Table (wherein each T may independently be replaced by U, and vice versa), and / or a sugar, nucleobase and / or internucleotide linkage modification and / or pattern thereof set forth in Table 1, and / or an additional chemical moiety (added to the oligonucleotide chain, e.g., a targeting moiety, lipid moiety, carbohydrate moiety, etc.) set forth in Table 1.

[0198] In some embodiments, the terms "complementary," "fully complementary," and "substantially complementary" can be used in reference to the degree of base identity between an oligonucleotide (e.g., a ds oligonucleotide targeting HSD17B13) base sequence and a target sequence (e.g., an HSD17B13 target sequence), as one of skill in the art would understand from the context of their use. Note that substituting U for T, or vice versa, generally does not change the amount of complementarity. As used herein, an oligonucleotide that is "substantially complementary" to a target sequence is largely or mostly complementary, but not 100%. In some embodiments, a substantially complementary sequence (e.g., a ds oligonucleotide targeting HSD17B13) has 1, 2, 3, 4, or 5 mismatches when aligned with its target sequence. In some embodiments, a ds oligonucleotide targeting HSD17B13 has a base sequence that is substantially complementary to an HSD17B13 target sequence. In some embodiments, the ds oligonucleotide targeting HSD17B13 has a base sequence substantially complementary to the complement of the sequence of a ds oligonucleotide targeting HSD17B13 disclosed herein. As one of skill in the art will understand, in some embodiments, the sequence of the oligonucleotide need not be 100% complementary to its target for the oligonucleotide to perform its function (e.g., knockdown of the target nucleic acid). When determining complementarity, typically, A and T (or U) are complementary nucleobases, and C and G are complementary nucleobases.

[0199] In some embodiments, the present disclosure provides ds oligonucleotides targeting HSD17B13 comprising a sequence found in an oligonucleotide listed in a Table. In some embodiments, the present disclosure provides ds oligonucleotides targeting HSD17B13 comprising a sequence found in an oligonucleotide listed in Table 1 (wherein one or more U's are independently and optionally replaced by T's, or vice versa). In some embodiments, the ds oligonucleotides targeting HSD17B13 may comprise at least one T and / or at least one U. In some embodiments, the present disclosure provides ds oligonucleotides targeting HSD17B13 comprising a sequence found in an oligonucleotide listed in a Table, wherein the sequence has greater than 50% identity to the sequence of the oligonucleotide listed in the Table. In some embodiments, the present disclosure provides ds oligonucleotides targeting HSD17B13 comprising the sequence of an oligonucleotide listed in Table 1. In some embodiments, the present disclosure provides ds oligonucleotides targeting HSD17B13, the base sequence of which is the sequence of an oligonucleotide disclosed in Table 1, where each T can be independently replaced by U, and vice versa. In some embodiments, the present disclosure provides ds oligonucleotides targeting HSD17B13, comprising a sequence found in an oligonucleotide in Table 1, where the oligonucleotide has the backbone linkage pattern, backbone chiral center pattern, and / or backbone phosphorus modification pattern of the same or another oligonucleotide in Table 1.

[0200] Among other things, the present disclosure provides a variety of ds oligonucleotides, each having a base sequence defined in Table 1 and elsewhere. In some embodiments, the present disclosure provides oligonucleotides having a base sequence that is, comprises, or comprises a portion of the base sequence of an oligonucleotide disclosed herein, e.g., in Table 1 herein (wherein each T may be independently replaced by U, and vice versa). In some embodiments, the present disclosure provides oligonucleotides having a base sequence that is, comprises, or comprises a portion of the base sequence of an oligonucleotide disclosed herein, e.g., in Table 1 (wherein each T may be independently replaced by U, and vice versa), where the oligonucleotide further comprises chemical modifications, stereochemistry, formatting, additional chemical moieties (e.g., targeting moieties, lipid moieties, carbohydrate moieties, etc.) and / or another structural feature as described herein.

[0201] In some embodiments, a "portion" (e.g., a portion of a base sequence or modification pattern) is at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 monomer units in length (e.g., a base sequence at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 bases in length). In some embodiments, a "portion" of a base sequence is at least 5 bases in length. In some embodiments, a "portion" of a base sequence is at least 10 bases in length. In some embodiments, a "portion" of a base sequence is at least 15 bases in length. In some embodiments, a "portion" of a base sequence is at least 16, 17, 18, 19, or 20 bases in length. In some embodiments, a "portion" of a base sequence is at least 20 bases in length. In some embodiments, the portion of the base sequence is 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or more contiguous (consecutive) bases. In some embodiments, the portion of the base sequence is 15 or more contiguous (consecutive) bases. In some embodiments, the portion of the base sequence is 16, 17, 18, 19, or 20 or more contiguous (consecutive) bases. In some embodiments, the portion of the base sequence is 20 or more contiguous (consecutive) bases.

[0202] In some embodiments, the present disclosure provides oligonucleotides (e.g., ds oligonucleotides targeting HSD17B13) having a base sequence of an oligonucleotide in Table 1 (wherein each T can be independently replaced by U, or vice versa). In some embodiments, the present disclosure provides ds oligonucleotides targeting HSD17B13 having a sequence of an oligonucleotide in Table 1, wherein the oligonucleotide has the ability to induce a decrease in the expression, level, and / or activity of the HSD17B13 gene or its gene product. As will be understood by one of skill in the art, in the provided base sequences, each U can be optionally and independently replaced by T, or vice versa, and the sequence can comprise a mixture of U and T. In some embodiments, C can be optionally and independently replaced by 5mC.

[0203] In some embodiments, the portion is a span of at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleotides in total. In some embodiments, the portion is a span of at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleotides in total that contains 0-3 mismatches. In some embodiments, the portion is a span of at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleotides in total with 0-3 mismatches, where spans with 0 mismatches are complementary and spans with one or more mismatches are substantially complementary, as non-limiting examples. In some embodiments, the bases include a portion that is characteristic of a nucleic acid (e.g., a gene) in that the portion is identical to or complementary to a portion of the nucleic acid or a transcript thereof, and is not identical to or complementary to a portion of any other nucleic acid (e.g., a gene) or transcript thereof within the same genome. In some embodiments, the portion is characteristic of human HSD17B13.

[0204] In some embodiments, provided oligonucleotides, e.g., ds oligonucleotides targeting HSD17B13, have a length of about 49, 45, 40, 30, 35, 25, or 23 total nucleotides or less, as described herein. In some embodiments where the 5' end of a sequence described herein begins with a U or a T, the U can be deleted and / or substituted with another base. In some embodiments, the oligonucleotide has a base sequence that is, includes, or comprises a portion of the base sequence of an oligonucleotide in a table (wherein each T can be independently replaced with a U, and vice versa), and has a format or portion of a format disclosed herein.

[0205] In some embodiments, oligonucleotides, such as ds oligonucleotides targeting HSD17B13, are stereorandom. In some embodiments, ds oligonucleotides targeting HSD17B13 are chirally controlled. In some embodiments, ds oligonucleotides targeting HSD17B13 are chirally pure (or "stereically pure," "stereochemically pure"), meaning that the oligonucleotide exists as a single stereoisomeric form (often as a single diastereoisomeric (or "diastereomer") form, since multiple chiral centers may exist in the oligonucleotide, e.g., at the bond phosphorus, sugar carbon, etc.). As will be understood by those skilled in the art, chirally pure oligonucleotides are separated from other stereoisomeric forms (to the extent that some impurities may exist, since chemical and biological processes, selectivity and / or purification, etc., are rarely, if ever, absolutely perfect). In chirally pure oligonucleotides, each chiral center is independently defined with respect to its configuration (for chirally pure oligonucleotides, each internucleotide linkage is independently stereorestricted or chiral controlled). In contrast to chirally controlled and chirally pure oligonucleotides containing sterically restricted linking phosphorus, racemic (or "stereorandom," "chirally uncontrolled") oligonucleotides containing chiral linking phosphorus, derived, for example, from conventional phosphoramidite oligonucleotide synthesis without stereochemical control during the coupling step in combination with conventional sulfurization (to generate stereorandom phosphorothioate internucleotide linkages), refer to an irregular mixture of various stereoisomers (typically diastereoisomers (or "diastereomers"), since multiple chiral centers are present in the oligonucleotide; for example, derived from conventional oligonucleotide preparations using reagents that contain no chiral elements other than those at the nucleotides and linking phosphorus). For example, in terms of A*A*A (where * is the phosphorothioate internucleotide linkage (containing the chiral linking phosphorus)), the preparation of racemic oligonucleotides can yield four diastereomers [2 2= 4, considering two chiral linking phosphorus, each of which can exist in either of two configurations (Sp or Rp): A*SA*SA, A*SA*RA, A*RA*SA, and A*RA*RA (*S represents the phosphorothioate internucleotide linkage of Sp, and *R represents the phosphorothioate internucleotide linkage of Rp). With respect to chirally pure oligonucleotides (e.g., A*SA*SA), the oligonucleotide exists in a single stereoisomeric form, and the oligonucleotide is separated from other stereoisomers (e.g., diastereomers A*SA*RA, A*RA*SA, and A*RA*RA).

[0206] In some embodiments, ds oligonucleotides targeting HSD17B13 contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more stereorandom internucleotide linkages (a mixture of Rp- and Sp-linked phosphorus at the internucleotide linkage, e.g., derived from conventional, non-chiral controlled oligonucleotide synthesis). In some embodiments, ds oligonucleotides targeting HSD17B13 contain one or more (e.g., 1-50, 1-40, 1-30, 1-25, 1-20, 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) chiral controlled internucleotide linkages (Rp- or Sp-linked phosphorus at the internucleotide linkage, e.g., derived from chiral controlled oligonucleotide synthesis). In some embodiments, the internucleotide linkages are phosphorothioate internucleotide linkages. In some embodiments, the internucleotide linkages are stereorandom phosphorothioate internucleotide linkages. In some embodiments, the internucleotide linkages are chiral-controlled phosphorothioate internucleotide linkages.

[0207] In particular, the present disclosure provides techniques for preparing chiral controlled (in some embodiments, stereochemically pure) oligonucleotides. In some embodiments, the oligonucleotides are stereochemically pure. In some embodiments, the oligonucleotides of the present disclosure have a chirality of about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, or about 5%, 10%, 15%, 20%, 25%, 30% , 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%, or at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99% pure. In some embodiments, the internucleotide linkages of the oligonucleotide are one or more (e.g., 1 to 50, 1 to 40, 1 to 30, 1 to 25, 1 to 20, 5 to 50, 5 to 40, 5 to 30, 5 to 25, 5 to 20, 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) chiral. In some embodiments, the oligonucleotides of the present disclosure, such as ds oligonucleotides targeting HSD17B13, comprise or consist of internucleotide linkages, each of which independently has a diastereomeric purity of at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%, typically at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%. CILwherein DS is a diastereomeric purity as described herein (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% or greater) and CIL is the number of chiral-controlling internucleotide linkages (e.g., 1 to 50, 1 to 40, 1 to 30, 1 to 25, 1 to 20, 5 to 50, 5 to 40, 5 to 30, 5 to 25, 5 to 20, 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 greater). In some embodiments, DS is 95% to 100%. In some embodiments, each internucleotide linkage is independently chiral-controlled, and CIL is the number of chiral-controlled internucleotide linkages.

[0208] By way of example, specific HSD17B13-targeting ds oligonucleotides, including specific example base sequences, nucleobase modifications and patterns thereof, sugar modifications and patterns thereof, internucleotide linkages and patterns thereof, linked phosphorus stereochemistry and patterns thereof, linkers, and / or additional chemical moieties, are set forth below in Table 1. In particular, oligonucleotides, such as those in Table 1, can be used to target HSD17B13 transcripts, e.g., to reduce levels of HSD17B13 transcripts and / or their products.

[0209] In certain exemplary embodiments, a ds oligonucleotide targeted to HSD17B13 of the present disclosure comprises a base sequence, nucleobase modifications and patterns thereof, sugar modifications and patterns thereof, internucleotide linkages and patterns thereof, linked phosphorus stereochemistry and patterns thereof, linker, and / or additional chemical moieties: WV-42589, WV-47139, WV-47159, WV-49590, or WV-49591. In certain exemplary embodiments, a ds oligonucleotide targeted to HSD17B13 of the present disclosure comprises a guide strand comprising a base sequence, nucleobase modifications and patterns thereof, sugar modifications and patterns thereof, internucleotide linkages and patterns thereof, linked phosphorus stereochemistry and patterns thereof, linker, and / or additional chemical moieties: WV-47139, WV-47159, WV-49590, or WV-49591. In certain exemplary embodiments, ds oligonucleotides targeting HSD17B13 of the present disclosure comprise a passenger strand comprising the base sequence, nucleobase modifications and their patterns, sugar modifications and their patterns, internucleotide linkages and their patterns, linked phosphorus stereochemistry and their patterns, linkers, and / or additional chemical moieties WV-42589. In certain exemplary embodiments, ds oligonucleotides targeting HSD17B13 of the present disclosure comprise a guide strand comprising the base sequence, nucleobase modifications and their patterns, sugar modifications and their patterns, internucleotide linkages and their patterns, linked phosphorus stereochemistry and their patterns, linkers, and / or additional chemical moieties WV-47139, WV-47159, WV-49590, or WV-49591, and a passenger strand comprising the base sequence, nucleobase modifications and their patterns, sugar modifications and their patterns, internucleotide linkages and their patterns, linked phosphorus stereochemistry and their patterns, linkers, and / or additional chemical moieties WV-42589.

[0210] In certain exemplary embodiments, the disclosed ds oligonucleotides targeting HSD17B13 comprise a guide strand comprising the base sequence, nucleobase modifications and their patterns, sugar modifications and their patterns, internucleotide linkages and their patterns, linked phosphorus stereochemistry and their patterns, linkers, and / or additional chemical moieties WV-47139, and a passenger strand comprising the base sequence, nucleobase modifications and their patterns, sugar modifications and their patterns, internucleotide linkages and their patterns, linked phosphorus stereochemistry and their patterns, linkers, and / or additional chemical moieties WV-42589. In certain exemplary embodiments, the disclosed ds oligonucleotides targeting HSD17B13 comprise a guide strand comprising WV-47139 and a passenger strand comprising WV-42589.

[0211] In certain exemplary embodiments, the disclosed ds oligonucleotides targeting HSD17B13 comprise a guide strand comprising the base sequence, nucleobase modifications and their patterns, sugar modifications and their patterns, internucleotide linkages and their patterns, linked phosphorus stereochemistry and their patterns, linkers, and / or additional chemical moieties WV-47159, and a passenger strand comprising the base sequence, nucleobase modifications and their patterns, sugar modifications and their patterns, internucleotide linkages and their patterns, linked phosphorus stereochemistry and their patterns, linkers, and / or additional chemical moieties WV-42589. In certain exemplary embodiments, the disclosed ds oligonucleotides targeting HSD17B13 comprise a guide strand comprising WV-47159 and a passenger strand comprising WV-42589.

[0212] In certain exemplary embodiments, the disclosed ds oligonucleotides targeting HSD17B13 comprise a guide strand comprising the base sequence, nucleobase modifications and patterns thereof, sugar modifications and patterns thereof, internucleotide linkages and patterns thereof, linked phosphorus stereochemistry and patterns thereof, linkers, and / or additional chemical moieties WV-49590, and a passenger strand comprising the base sequence, nucleobase modifications and patterns thereof, sugar modifications and patterns thereof, internucleotide linkages and patterns thereof, linked phosphorus stereochemistry and patterns thereof, linkers, and / or additional chemical moieties WV-42589. In certain exemplary embodiments, the disclosed ds oligonucleotides targeting HSD17B13 comprise a guide strand comprising WV-49590 and a passenger strand comprising WV-42589.

[0213] In certain exemplary embodiments, the disclosed ds oligonucleotides targeting HSD17B13 comprise a guide strand comprising the base sequence, nucleobase modifications and patterns thereof, sugar modifications and patterns thereof, internucleotide linkages and patterns thereof, linked phosphorus stereochemistry and patterns thereof, linkers, and / or additional chemical moieties WV-49591, and a passenger strand comprising the base sequence, nucleobase modifications and patterns thereof, sugar modifications and patterns thereof, internucleotide linkages and patterns thereof, linked phosphorus stereochemistry and patterns thereof, linkers, and / or additional chemical moieties WV-42589. In certain exemplary embodiments, the disclosed ds oligonucleotides targeting HSD17B13 comprise a guide strand comprising WV-49591 and a passenger strand comprising WV-42589.

[0214] [Table 1]

[0215] [Table 2]

[0216] [Table 3]

[0217]

Table 4

[0218]

Table 5

[0219]

Table 6

[0220]

Table 7

[0221]

Table 8

[0222]

Table 9

[0223]

Table 10

[0224]

Table 11

[0225]

Table 12

[0226]

Table 13

[0227]

Table 14

[0228]

Table 15

[0229] Table 16

[0230] Table 17

[0231] Table 18

[0232] Table 19

[0233] Table 20

[0234] Table 21

[0235] Table 22

[0236] Table 23

[0237] Table 24

[0238] Table 25

[0239] Table 26

[0240] Table 27

[0241] Table 28

[0242] Table 29

[0243] Table 30

[0244] Table 31

[0245] Table 32

[0246] Table 33

[0247] Table 34

[0248] Table 35

[0249] Table 36

[0250] Table 37

[0251] Table 38

[0252] Table 39

[0253] Table 40

[0254] Table 41

[0255] Table 42

[0256] Table 43

[0257] Table 44

[0258] Table 45

[0259] Table 46

[0260] Table 47

[0261] Table 48

[0262] Table 49

[0263]

Table 50

[0264] Table 51

[0265] Table 52

[0266] Table 53

[0267] Table 54

[0268] Table 55

[0269] Table 56

[0270] Table 57

[0271] Table 58

[0272] Table 59

[0273] Table 60

[0274] Table 61

[0275] Table 62

[0276] Table 63

[0277] Table 64

[0278] Table 65

[0279] Table 66

[0280] Table 67

[0281] Table 68

[0282] Table 69

[0283] Table 70

[0284] Table 71

[0285] Table 72

[0286] Table 73

[0287] Table 74

[0288] Table 75

[0289] Table 76

[0290] Table 77

[0291] Table 78

[0292] Table 79

[0293] Table 80

[0294] Table 81

[0295] Table 82

[0296] Table 83

[0297] Table 84

[0298] Table 85

[0299] Table 86

[0300] Table 87

[0301] Table 88

[0302] Table 89

[0303] Table 90

[0304]

Table 91

[0305] Table 92

[0306] Table 93

[0307] Table 94

[0308] Table 95

[0309] Table 96

[0310] [Table 97]

[0311] [Table 98]

[0312] [Table 99]

[0313] [Table 100]

[0314] Note: Descriptions, base sequences, and stereochemistry / linkages can be categorized into multiple series in Table 1 depending on their length. Unless otherwise specified, all oligonucleotides in Table 1 are single-stranded. As will be understood by those skilled in the art, nucleoside units are unmodified unless otherwise indicated (e.g., r, m, etc.), contain unmodified nucleobases and 2'-deoxy sugars, linkages are natural phosphate linkages unless otherwise indicated, and acidic / basic groups may independently be present in their salt form. When no sugar is specified, the sugar is the natural DNA sugar, and when no internucleotide linkage is specified, the internucleotide linkage is the natural phosphate linkage. Moieties and Modifications: m:2'-OMe; f:2′-F; O, PO: phosphodiester (phosphate). It can be a bond or a terminal group (or a component thereof), for example, the bond between the linker and the oligonucleotide chain, the internucleotide bond (natural phosphate bond), etc. A phosphodiester is usually indicated with "O" in the stereochemistry / bonding column and is usually not labeled in the description column (if it is a terminal group, for example, the 5'-terminal group, it is indicated in the description and usually not in the stereochemistry / bonding column); if a bond is not indicated in the description column, it is usually a phosphodiester unless otherwise indicated. Note that a phosphate bond between a linker (e.g., L001) and an oligonucleotide chain may not be labeled in the description column, but may be indicated with "O" in the stereochemistry / bonding column; *, PS: phosphorothioate, which can be an end group (if it is an end group, e.g., the 5' end group, this is indicated in the description and usually not in the stereochemistry / bonding) or a bond, e.g., a bond between a linker (e.g., L001) and an oligonucleotide chain, an internucleotide bond (phosphorothioate internucleotide bond), etc.; R, Rp: phosphorothioate in the Rp configuration. Note that *R in the description indicates a single phosphorothioate bond in the Rp configuration; S, Sp: phosphorothioate in Sp configuration. Note that *S in the description indicates a single phosphorothioate bond in Sp configuration; X: stereorandom phosphorothioate; n001: [ka] nX:stereorandom n001; nR or n001R: n001 in Rp configuration; nS or n001S: n001 in Sp configuration; n009: [ka] nX:stereorandom n009; nR or n009R: n009 in Rp configuration; nS or n009S: n009 in Sp configuration; n031: [ka] nX:stereorandom n031; nR or n031R: n031 in Rp configuration; nS or n031S: n031 in Sp configuration; n033: [ka] nX:stereorandom n033; nR or n033R: n033 in Rp configuration; nS or n033S: n033 in Sp configuration; n037: [ka] nX:stereorandom n037; nR or n037R: n037 in Rp configuration; nS or n037S: n037 in Sp configuration; n046: [ka] nX:stereorandom n046; nR or n046R: n046 in Rp configuration; nS or n046S: n046 in Sp configuration; n047: [ka] nX:stereorandom n047; nR or n047R: n047 in Rp configuration; nS or n047S: n047 in Sp configuration; n025: [ka] nX:stereorandom n025; nR or n025R: n025 in Rp configuration; nS or n025S: n025 in Sp configuration; n054: [ka] nX:stereorandom n054; nR or n054R: n054 in Rp configuration; nS or n054S: n054 in Sp configuration; n055: [ka] nX:stereorandom n055; nR or n055R: n055 in Rp configuration; nS or n055S: n055 in Sp configuration; n026: [ka] nX:stereorandom n001; nR or n026R: n026 in Rp configuration; nS or n026S: n026 in Sp configuration; n004: [ka] nX:stereorandom n004; nR or n004R: n004 in Rp configuration; nS or n004S: n004 in Sp configuration; n003: [ka] nX:stereorandom n003; nR or n003R: n003 in Rp configuration; nS or n003S: n003 in Sp configuration; n008: [ka] nX:stereorandom n008; nR or n008R: n008 in Rp configuration; nS or n008S: n008 in Sp configuration; n029: [ka] nX:stereorandom n029; nR or n029R: n029 in Rp configuration; nS or n029S: n029 in Sp configuration; n021: [ka] nX:stereorandom n021; nR or n021R: n021 in Rp configuration; nS or n021S: n021 in Sp configuration; n006: [ka] nX:stereorandom n006; nR or n006R: n006 in Rp configuration; nS or n006S: n006 in Sp configuration; n020: [ka] nX:stereorandom n020; nR or n020R: n020 in Rp configuration; nS or n020S: n020 in Sp configuration; X: stereorandom phosphorothioate; [ka] [ka] [ka] [ka] (wherein -C(O)- is attached to nitrogen); [ka] i.e., morpholine carbamate internucleotide linkage [ka] [ka] [ka] L001: -NH-(CH2)6- linker (C6 linker, C6 amine linker, or C6 amino linker) linked to Mod (e.g., Mod001) via -NH- and, in the case of WV-38061, to the 5'-end of the oligonucleotide chain via a phosphate bond (O or PO). For example, in WV-38061, L001 is linked to Mod001 via -NH- (forming an amide group -C(O)-NH-) and linked to the oligonucleotide chain via a phosphate bond (O). L010: [ka] In some embodiments, when L010 is present in the middle of an oligonucleotide, it is attached to an internucleotide linkage as another sugar (e.g., a DNA sugar), e.g., the 5'-carbon is linked to another unit (e.g., the 3' of the sugar) and the 3'-carbon is linked to another unit (e.g., the 5'-carbon of the carbon), independently, e.g., via a linkage (e.g., a phosphate linkage (O or PO) or a phosphorothioate linkage (which may be chiral non-controlled or chiral controlled (Sp or Rp))); L012: -CH2CH2OCH2CH2OCH2CH2-. When L012 is present in the middle of an oligonucleotide, each of its two ends is independently linked to an internucleotide bond (e.g., a phosphate bond (O or PO) or a phosphorothioate bond (which may be chiral non-controlled or chiral controlled (Sp or Rp))); L022: [ka] where L022 is linked to the rest of the molecule via the phosphate unless otherwise indicated; L023: HO-(CH2)6-, where CH2 is linked to the rest of the molecule via a phosphate unless otherwise indicated, e.g., in WV-42644 (where the O in OnRnRnRnRSSSSSSSSSSSSSSSSnRSSSSSnRSSnR indicates the phosphate bond linking L023 to the rest of the molecule); L025: [ka] wherein the -CH2- linkage serves as the C5 linkage of the sugar (e.g., DNA sugar) and is linked to another unit (e.g., the 3' of the sugar), and the linkage on the ring serves as the C3 linkage and is linked to another unit (the 5'-carbon of the carbon), each of which is independently via, for example, a bond (e.g., a phosphate bond (O or PO) or a phosphorothioate bond (which may be chiral non-controlled or chiral controlled (Sp or Rp)). When L025 is at the 5' end without any modification, the -CH2- linkage is attached to -OH. For example, L025L025L025- in various oligonucleotides is [ka] (which may exist as various salt forms) and is linked to the 5'-carbon of an oligonucleotide chain via a specified bond (e.g., a phosphate bond (O or PO) or a phosphorothioate bond (which may be chiral non-controlled or chiral controlled (Sp or Rp))); L016: [ka] where L016, unless otherwise indicated, is linked to the remainder of the molecule via the phosphate, and L016 is utilized with n001 to form L016n001, which has the following structure: [ka]

[0315] Length of double-stranded oligonucleotide As will be appreciated by those skilled in the art, ds oligonucleotides targeting HSD17B13 can vary in length to provide the properties and / or activities required for various applications. Numerous techniques for evaluating, selecting, and / or optimizing the length of oligonucleotides are available in the art and can be utilized in accordance with the present disclosure. As demonstrated herein, in many embodiments, ds oligonucleotides targeting HSD17B13 are of a length suitable for hybridizing to their targets and reducing the levels of those targets and / or their encoded products. In some embodiments, the oligonucleotides are sufficiently long to recognize the target nucleic acid (e.g., HSD17B13 mRNA). In some embodiments, the oligonucleotides are sufficiently long to distinguish the target nucleic acid from other nucleic acids (e.g., nucleic acids having non-HSD17B13 base sequences) and reduce off-target effects. In some embodiments, ds oligonucleotides targeting HSD17B13 are sufficiently short to reduce manufacturing or production complexity and lower product costs.

[0316] In some embodiments, the base sequence of the oligonucleotide is about 10 to 500 nucleobases in length. In some embodiments, the base sequence is about 10 to 500 nucleobases in length. In some embodiments, the base sequence is about 10 to 50 nucleobases in length. In some embodiments, the base sequence is about 15 to 50 nucleobases in length. In some embodiments, the base sequence is about 15 to about 30 nucleobases in length. In some embodiments, the base sequence is about 10 to about 25 nucleobases in length. In some embodiments, the base sequence is about 15 to about 22 nucleobases in length. In some embodiments, the base sequence is about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleobases in length. In some embodiments, the base sequence is about 18 nucleobases in length. In some embodiments, the base sequence is about 19 nucleobases in length. In some embodiments, the base sequence is about 20 nucleobases in length. In some embodiments, the base sequence is about 21 nucleobases in length. In some embodiments, the base sequence is about 22 nucleobases in length. In some embodiments, the base sequence is about 23 nucleobases in length. In some embodiments, the base sequence is about 24 nucleobases in length. In some embodiments, the base sequence is about 25 nucleobases in length. In some embodiments, each nucleobase is an optionally substituted A, T, C, G, U, or an optionally substituted tautomer of A, T, C, G, or U.

[0317] Internucleotide linkages in double-stranded oligonucleotides In some embodiments, ds oligonucleotides targeting HSD17B13 comprise base modifications, sugar modifications, and / or internucleotide linkage modifications. Various internucleotide linkages can be used to link nucleobase-containing units (e.g., nucleosides). In some embodiments, ds oligonucleotides targeting HSD17B13 comprise both one or more modified internucleotide linkages and one or more natural phosphate linkages. As is well known to those skilled in the art, natural phosphate linkages are widely found in natural DNA and RNA molecules, have the structure -OP(O)(OH)O-, and link sugars in nucleosides in DNA and RNA. They can exist in various salt forms, for example, at physiological pH (about 7.4), and natural phosphate linkages are those in which the anion is -OP(O)(O - )O-. A modified internucleotide linkage or non-natural phosphate linkage is an internucleotide linkage that is not a natural phosphate linkage or its salt form. A modified internucleotide linkage may also exist in a salt form depending on its structure. For example, as will be understood by those skilled in the art, a phosphorothioate internucleotide linkage having the structure -OP(O)(SH)O- can exist in various salt forms, for example, at physiological pH (about 7.4), with the anion being -OP(O)(S - )O-.

[0318] In some embodiments, the oligonucleotide comprises an internucleotide linkage that is a modified internucleotide linkage, for example, a phosphorothioate, phosphorodithioate, methylphosphonate, phosphoramidate, thiophosphate, 3'-thiophosphate, or 5'-thiophosphate.

[0319] In some embodiments, the modified internucleotide linkage is a chiral internucleotide linkage comprising a chiral linking phosphorus. In some embodiments, the chiral internucleotide linkage is a phosphorothioate linkage. In some embodiments, the chiral internucleotide linkage is a non-negatively charged internucleotide linkage. In some embodiments, the chiral internucleotide linkage is a neutral internucleotide linkage. In some embodiments, the chiral internucleotide linkage is chiral controlled with respect to its chiral linking phosphorus. In some embodiments, the chiral internucleotide linkage is stereochemically pure with respect to its chiral linking phosphorus. In some embodiments, the chiral internucleotide linkage is not chiral controlled. In some embodiments, the pattern of backbone chiral centers comprises or consists of chiral controlled internucleotide linkage positions and linking phosphorus configurations (Rp or Sp), and achiral internucleotide linkage (e.g., natural phosphate linkage) positions.

[0320] In certain embodiments, the internucleotide linkage comprises a P-modification, which is a modification at the linked phosphorus. In certain embodiments, the modified internucleotide linkage does not contain phosphorus, as in the case of peptide nucleic acids (PNAs), but is a moiety that serves to link two sugars or two moieties, each independently comprising a nucleobase.

[0321] In certain embodiments, the ds oligonucleotide has a modified internucleotide linkage, e.g., a structure of Formula I, Ia, Ib, or Ic, as described herein and / or in WO 2018 / 022473, WO 2018 / 098264, WO 2018 / 2230, each of which and / or each internucleotide linkage (e.g., of Formula I, Ia, Ib, Ic, etc.) is independently incorporated herein by reference. 56, WO 2018 / 223073, WO 2018 / 223081, WO 2018 / 237194, WO 2019 / 032607, WO 2019 / 055951, WO 2019 / 075357, WO 2019 / 200185, WO 2019 / 217784, and / or WO 2019 / 032612. In certain embodiments, the modified internucleotide linkage is a chiral internucleotide linkage. In certain embodiments, the modified internucleotide linkage is a phosphorothioate internucleotide linkage.

[0322] In certain embodiments, the modified internucleotide linkage is a non-negatively charged internucleotide linkage. In certain embodiments, provided ds oligonucleotides comprise one or more non-negatively charged internucleotide linkages. In certain embodiments, the non-negatively charged internucleotide linkage is a positively charged internucleotide linkage. In certain embodiments, the non-negatively charged internucleotide linkage is a neutral internucleotide linkage. In certain embodiments, the present disclosure provides ds oligonucleotides comprising one or more neutral internucleotide linkages. In certain embodiments, the non-negatively charged internucleotide linkage is a non-negatively charged internucleotide linkage as described herein and / or U.S. Patent No. 939,939, each of which is independently incorporated herein by reference. 4333, U.S. Patent No. 9744183, U.S. Patent No. 9605019, U.S. Patent No. 9982257, U.S. Patent Application Publication No. 20170037399, U.S. Patent Application Publication No. 20180216108, U.S. Patent Application Publication No. 20180216107, U.S. Patent No. 9598458, WO 2017 / 062862, WO 2018 / 067973, WO 2017 / 160741, WO 2017 / 160741 International Publication No. 2017 / 192679, International Publication No. 2017 / 210647, International Publication No. 2018 / 098264, International Publication No. 2018 / 022473, International Publication No. 2018 / 223056, International Publication No. 2018 / 223073, International Publication No. 2018 / 223081, International Publication No. 2018 / 237194, International Publication No. 2019 / 032607, International Publication No. 2019 / 032612, International Publication No. 2019 / 055 951, WO 2019 / 075357, WO 2019 / 200185, WO 2019 / 217784 and / or WO 2019 / 032612, or a salt form thereof.

[0323] In certain embodiments, non-negatively charged internucleotide linkages can improve delivery and / or activity (e.g., adenosine editing activity).

[0324] In certain embodiments, the modified internucleotide linkage (e.g., a non-negatively charged internucleotide linkage) comprises an optionally substituted triazolyl. In certain embodiments, the modified internucleotide linkage (e.g., a non-negatively charged internucleotide linkage) comprises an optionally substituted alkynyl. In certain embodiments, the modified internucleotide linkage comprises a triazole or alkyne moiety. In certain embodiments, the triazole moiety, e.g., a triazolyl group, is optionally substituted. In some embodiments, the triazole moiety, e.g., a triazolyl group, is substituted. In certain embodiments, the triazole moiety is unsubstituted. In certain embodiments, the modified internucleotide linkage comprises an optionally substituted cyclic guanidine moiety. In certain embodiments, the modified internucleotide linkage is [ka] and optionally chiral controlled, wherein R 1 is -L-R', where L is L as described herein. B and R' is as described herein. In certain embodiments, each R 1 is independently R'. In certain embodiments, each R' is independently R. In certain embodiments, two R 1 is R and together form a ring as described herein. In certain embodiments, two R on two different nitrogen atoms 1 is R and taken together to form a ring as described herein. In certain embodiments, R 1 is independently an optionally substituted C as described herein. 1~6 In certain embodiments, R1 is methyl. In certain embodiments, two R' on the same nitrogen atom are R and together form a ring as described herein. In certain embodiments, the modified internucleotide linkage is [ka] and optionally chiral controlled. In certain embodiments, [ka] teeth, [ka] In certain embodiments, the modified internucleotide linkage comprises an optionally substituted cyclic guanidine moiety: [ka] wherein W is O or S. In certain embodiments, W is O. In certain embodiments, W is S. In certain embodiments, the non-negatively charged internucleotide linkage is stereochemically controlled.

[0325] In certain embodiments, the non-negatively charged or neutral internucleotide linkage is an internucleotide linkage comprising a triazole moiety. In some embodiments, the internucleotide linkage comprising a triazole moiety (e.g., an optionally substituted triazolyl group) is [ka] In some embodiments, the internucleotide linkage comprising a triazole moiety has the structure: [ka] In some embodiments, the internucleotide linkage comprising a triazole moiety has the structure: [ka] where W is O or S. In some embodiments, an internucleotide linkage comprising an alkyne moiety (e.g., an optionally substituted alkynyl group) has the formula: [ka] wherein W is O or S. In some embodiments, the internucleotide linkage, e.g., a non-negatively charged internucleotide linkage, a neutral internucleotide linkage, comprises a cyclic guanidine moiety. In some embodiments, the internucleotide linkage comprising a cyclic guanidine moiety is [ka] In some embodiments, the non-negatively charged or neutral internucleotide linkage has the structure: [ka] wherein W is O or S. In certain embodiments, the internucleotide linkage, e.g., a non-negatively charged internucleotide linkage, a neutral internucleotide linkage, comprises a cyclic guanidine moiety. In certain embodiments, the internucleotide linkage comprising a cyclic guanidine moiety is [ka] In certain embodiments, the non-negatively charged or neutral internucleotide linkage has the structure: [ka] wherein W is O or S.

[0326] In certain embodiments, the internucleotide linkage is [ka] In certain embodiments, the internucleotide linkage comprises a Tmg group, [ka] ("Tmg internucleotide linkage"). In certain embodiments, neutral internucleotide linkages include PNA and PMO internucleotide linkages, and Tmg internucleotide linkages.

[0327] In certain embodiments, a non-negatively charged internucleotide linkage has a structure such as Formula I, Ia, Ib, Ic, In-1, In-2, In-3, In-4, II, II-a-1, II-a-2, II-b-1, II-b-2, II-c-1, II-c-2, II-d-1, II-d-2, or a salt form thereof. In certain embodiments, a non-negatively charged internucleotide linkage comprises an optionally substituted 3-20-membered heterocyclyl or heteroaryl group having 1-10 heteroatoms. In certain embodiments, a non-negatively charged internucleotide linkage comprises an optionally substituted 3-20-membered heterocyclyl or heteroaryl group having 1-10 heteroatoms, wherein at least one heteroatom is nitrogen. In certain embodiments, such heterocyclyl or heteroaryl groups are 5-membered rings. In certain embodiments, such heterocyclyl or heteroaryl groups are 6-membered rings.

[0328] In certain embodiments, the non-negatively charged internucleotide linkage comprises an optionally substituted 5-20-membered heterocyclyl or heteroaryl group having 1-10 heteroatoms. In certain embodiments, the non-negatively charged internucleotide linkage comprises an optionally substituted 5-20-membered heterocyclyl or heteroaryl group having 1-10 heteroatoms, wherein at least one heteroatom is nitrogen. In certain embodiments, the non-negatively charged internucleotide linkage comprises an optionally substituted 5-6-membered heterocyclyl or heteroaryl group having 1-4 heteroatoms, wherein at least one heteroatom is nitrogen. In certain embodiments, the non-negatively charged internucleotide linkage comprises an optionally substituted 5-membered heterocyclyl or heteroaryl group having 1-4 heteroatoms, wherein at least one heteroatom is nitrogen. In certain embodiments, the non-negatively charged internucleotide linkage comprises an optionally substituted 5-membered heterocyclyl or heteroaryl group having 1-4 heteroatoms, wherein at least one heteroatom is nitrogen. In certain embodiments, the heteroaryl group is directly attached to the linking phosphorus.

[0329] In certain embodiments, the non-negatively charged internucleotide linkage comprises an optionally substituted 5- to 20-membered heterocyclyl group having 1 to 10 heteroatoms. In certain embodiments, the non-negatively charged internucleotide linkage comprises an optionally substituted 5- to 20-membered heterocyclyl group having 1 to 10 heteroatoms, wherein at least one heteroatom is nitrogen. In certain embodiments, the non-negatively charged internucleotide linkage comprises an optionally substituted 5- to 6-membered heterocyclyl group having 1 to 4 heteroatoms, wherein at least one heteroatom is nitrogen. In certain embodiments, the non-negatively charged internucleotide linkage comprises an optionally substituted 5-membered heterocyclyl group having 1 to 4 heteroatoms, wherein at least one heteroatom is nitrogen. In certain embodiments, at least two heteroatoms are nitrogen. In some embodiments, the non-negatively charged internucleotide linkage comprises an optionally substituted triazolyl group. In some embodiments, the non-negatively charged internucleotide linkage comprises an unsubstituted triazolyl group, for example. [ka] In some embodiments, the non-negatively charged internucleotide linkage comprises a substituted triazolyl group, e.g. [ka] Includes.

[0330] In certain embodiments, the heterocyclyl group is directly attached to the phosphorus bond. In certain embodiments, the heterocyclyl group is attached to the phosphorus bond via a linker, e.g., via =N- when the heterocyclyl group is part of a guanidine moiety that is directly attached to the phosphorus bond via =N-. In certain embodiments, the non-negatively charged internucleotide linkage is optionally substituted. [ka] In certain embodiments, the non-negatively charged internucleotide linkage comprises a substituted [ka] In certain embodiments, the non-negatively charged internucleotide linkage comprises a group: [ka] group (wherein each R 1 is independently -LR. In certain embodiments, each R 1 are independently optionally substituted C 1~6 In certain embodiments, each R 1 is independently methyl.

[0331] In certain embodiments, the modified internucleotide linkage, e.g., the non-negatively charged internucleotide linkage, comprises a triazole or alkyne moiety, each of which is optionally substituted. In certain embodiments, the modified internucleotide linkage comprises a triazole moiety. In certain embodiments, the modified internucleotide linkage comprises an unsubstituted triazole moiety. In certain embodiments, the modified internucleotide linkage comprises a substituted triazole moiety. In certain embodiments, the modified internucleotide linkage comprises an alkyl moiety. In certain embodiments, the modified internucleotide linkage comprises an optionally substituted alkynyl group. In certain embodiments, the modified internucleotide linkage comprises an unsubstituted alkynyl group. In certain embodiments, the modified internucleotide linkage comprises a substituted alkynyl group. In certain embodiments, the alkynyl group is directly attached to the phosphorus linkage.

[0332] In certain embodiments, ds oligonucleotides contain different types of internucleotide phosphorus linkages. In certain embodiments, chiral controlled oligonucleotides contain at least one natural phosphate linkage and at least one modified (non-natural) internucleotide linkage. In certain embodiments, ds oligonucleotides contain at least one natural phosphate linkage and at least one phosphorothioate. In certain embodiments, ds oligonucleotides contain at least one non-negatively charged internucleotide linkage. In certain embodiments, ds oligonucleotides contain at least one natural phosphate linkage and at least one non-negatively charged internucleotide linkage. In certain embodiments, ds oligonucleotides contain at least one phosphorothioate internucleotide linkage and at least one non-negatively charged internucleotide linkage. In certain embodiments, ds oligonucleotides contain at least one phosphorothioate internucleotide linkage, at least one natural phosphate linkage, and at least one non-negatively charged internucleotide linkage. In certain embodiments, the ds oligonucleotide comprises one or more, e.g., 1-50, 1-40, 1-30, 1-20, 1-15, 1-10, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more, non-negatively charged internucleotide linkages. In certain embodiments, the non-negatively charged internucleotide linkages are not negatively charged in that, in aqueous solution at a given pH, less than 50%, 40%, 40%, 30%, 20%, 10%, 5%, or 1% of the internucleotide linkages are present in the negatively charged salt form. In certain embodiments, the pH is about pH 7.4. In certain embodiments, the pH is about pH 4-9. In certain embodiments, the percentage is less than 10%. In certain embodiments, the percentage is less than 5%. In certain embodiments, the percentage is less than 1%. In certain embodiments, the internucleotide linkages are non-negatively charged in that the neutral form of the internucleotide linkage does not have a pKa in water of about 1, 2, 3, 4, 5, 6, or 7 or less. In certain embodiments, none of the pKas is 7 or less.In certain embodiments, none of the pKas is 6 or less. In certain embodiments, none of the pKas is 5 or less. In certain embodiments, none of the pKas is 4 or less. In certain embodiments, none of the pKas is 3 or less. In certain embodiments, none of the pKas is 2 or less. In certain embodiments, none of the pKas is 1 or less. In certain embodiments, the pKa of the neutral form of the internucleotide linkage can be represented by the pKa of the neutral form of a compound having the structure CH3-internucleotide linkage-CH3. For example, the pKa of the neutral form of the internucleotide linkage having the structure of Formula I is: [ka] (wherein each of X, Y, and Z is independently —O—, —S—, or —N(R′)—; L is L B and R 1 is -L-R'), [ka] pKa of [ka] In certain embodiments, the non-negatively charged internucleotide linkage is a neutral internucleotide linkage. In certain embodiments, the non-negatively charged internucleotide linkage is a positively charged internucleotide linkage. In certain embodiments, the non-negatively charged internucleotide linkage comprises a guanidine moiety. In certain embodiments, the non-negatively charged internucleotide linkage comprises a heteroaryl base moiety. In certain embodiments, the non-negatively charged internucleotide linkage comprises a triazole moiety. In certain embodiments, the non-negatively charged internucleotide linkage comprises an alkynyl moiety.

[0333] In certain embodiments, the neutral or non-negatively charged internucleotide linkage is selected from the group consisting of those described in U.S. Pat. Nos. 9,394,333, 9,744,183, 9,605,019, 9,982,257, U.S. Patent Application Publication No. 20170037399, U.S. Patent Application Publication No. 20180216108, and the like, each of which is incorporated by reference herein. U.S. Patent Application Publication No. 20180216107, U.S. Patent No. 9,598,458, WO 2017 / 062862, WO 2018 / 067973, WO 2017 / 160741, WO 2017 / 192679, WO 2017 / 210647, WO 2018 / 098264, WO 2018 / 022473, WO 2018 / 22 3056, WO 2018 / 223073, WO 2018 / 223081, WO 2018 / 237194, WO 2019 / 032607, WO 2019 / 032612, WO 2019 / 055951, WO 2019 / 075357, WO 2019 / 200185, WO 2019 / 217784 and / or The structure of the neutral or non-negatively charged internucleotide bond is any of those described in International Publication Nos. 2019 / 032612, 2607, 2019032612, 2019 / 055951, 2019 / 075357, 2019 / 200185, 2019 / 217784, and / or 2019 / 032612.

[0334] In certain embodiments, each R' is independently an optionally substituted C 1~6 In certain embodiments, each R' is independently an optionally substituted C 1~6 In certain embodiments, each R' is independently -CH3. In certain embodiments, each R S is -H.

[0335] In certain embodiments, the non-negatively charged internucleotide linkage is [ka] In certain embodiments, the non-negatively charged internucleotide linkage has the structure: [ka] In certain embodiments, the non-negatively charged internucleotide linkage has the structure: [ka] In some embodiments, the non-negatively charged internucleotide linkage has the structure: [ka] In some embodiments, the non-negatively charged internucleotide linkage has the structure: [ka] In some embodiments, the non-negatively charged internucleotide linkage has the structure: [ka] In some embodiments, the non-negatively charged internucleotide linkage has the structure: [ka] In some embodiments, the non-negatively charged internucleotide linkage has the structure: [ka] In some embodiments, the non-negatively charged internucleotide linkage has the structure: [ka] In some embodiments, the non-negatively charged internucleotide linkage has the structure: [ka] In some embodiments, the non-negatively charged internucleotide linkage has the structure: [ka] In some embodiments, the non-negatively charged internucleotide linkage has the structure: [ka] In some embodiments, W is O. In some embodiments, W is S. In some embodiments, the neutral internucleotide linkage is a non-negatively charged internucleotide linkage as described above.

[0336] In certain embodiments, provided ds oligonucleotides comprise one or more internucleotide linkages of formula I, Ia, Ib, Ic, In-1, In-2, In-3, In-4, II, II-a-1, II-a-2, II-b-1, II-b-2, II-c-1, II-c-2, II-d-1, or II-d-2, which are represented by the formula I, Ia, Ib, Ic, In-1, In-2, In-3, In-4, II, II-a-1, II-a-2, II-b-1, II-b-2, I I-c-1, II-c-2, II-d-1 or II-d-2 or salt forms thereof are independently incorporated by reference herein, including U.S. Pat. No. 9,394,333, U.S. Pat. No. 9,744,183, U.S. Pat. No. 9,605,019, U.S. Pat. No. 9,982,257, U.S. Patent Application Publication No. 20170037399, U.S. Patent Application Publication No. 20180216108, U.S. Patent Application Publication No. 20180216107, U.S. Pat. No. 9,598,458, WO 2017 / 062862 ... International Publication No. 2018 / 067973, International Publication No. 2017 / 160741, International Publication No. 2017 / 192679, International Publication No. 2017 / 210647, International Publication No. 2018 / 098264, International Publication No. 2018 / 022473, International Publication No. 2018 / 223056, International Publication No. 2018 / 223073, International Publication No. 2018 / 223081, International Publication No. 2018 / 237194, International Publication No. 2019 / 032607, International Publication No. 2019 / 032612, International Publication No. and / or WO 2019 / 055951, WO 2019 / 075357, WO 2019 / 200185, WO 2019 / 217784 and / or WO 2019 / 032612, WO 2607, WO 2019 / 032612, WO 2019 / 055951, WO 2019 / 075357, WO 2019 / 200185, WO 2019 / 217784 and / or WO 2019 / 032612.

[0337] In certain embodiments, a ds oligonucleotide comprises a neutral internucleotide linkage and a chiral-controlled internucleotide linkage. In certain embodiments, a ds oligonucleotide comprises a neutral internucleotide linkage and a chiral-controlled internucleotide linkage that is not a neutral internucleotide linkage. In certain embodiments, a ds oligonucleotide comprises a neutral internucleotide linkage and a chiral-controlled phosphorothioate internucleotide linkage. In certain embodiments, the present disclosure provides a ds oligonucleotide comprising one or more non-negatively charged internucleotide linkages and one or more phosphorothioate internucleotide linkages, wherein each phosphorothioate internucleotide linkage in the oligonucleotide is independently a chiral-controlled internucleotide linkage. In certain embodiments, the present disclosure provides a ds oligonucleotide comprising one or more neutral internucleotide linkages and one or more phosphorothioate internucleotide linkages, wherein each phosphorothioate internucleotide linkage in the ds oligonucleotide is independently a chiral-controlled internucleotide linkage. In certain embodiments, the ds oligonucleotide comprises at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more chiral-controlled phosphorothioate internucleotide linkages. In certain embodiments, the non-negatively charged internucleotide linkages are chiral-controlled. In certain embodiments, the non-negatively charged internucleotide linkages are not chiral-controlled. In certain embodiments, the neutral internucleotide linkages are chiral-controlled. In certain embodiments, the neutral internucleotide linkages are not chiral-controlled.

[0338] Without wishing to be bound by any particular theory, the present disclosure recognizes that neutral internucleotide linkages may be more hydrophobic than phosphorothioate internucleotide linkages (PS), which may be more hydrophobic than natural phosphate linkages (PO). Typically, unlike PS or PO, neutral internucleotide linkages have a lower charge. Without wishing to be bound by any particular theory, the present disclosure recognizes that incorporating one or more neutral internucleotide linkages into a ds oligonucleotide may increase the ability of the ds oligonucleotide to be taken up by cells and / or escape from endosomes. Without wishing to be bound by any particular theory, the present disclosure recognizes that incorporating one or more neutral internucleotide linkages may be used to adjust the melting temperature of a duplex formed between a ds oligonucleotide and its target nucleic acid.

[0339] Without wishing to be bound by any particular theory, the present disclosure recognizes that incorporating one or more non-negatively charged internucleotide linkages, e.g., neutral internucleotide linkages, into a ds oligonucleotide can increase the ability of the ds oligonucleotide to mediate a function such as targeted adenosine editing.

[0340] As will be appreciated by those skilled in the art, internucleotide linkages such as natural phosphate linkages and those of formula I, Ia, Ib, Ic, In-1, In-2, In-3, In-4, II, II-a-1, II-a-2, II-b-1, II-b-2, II-c-1, II-c-2, II-d-1, II-d-2, or salt forms thereof, are typically represented by formula I, Ia, Ib, Ic, In-1, In-2, In-3, In-4, II, II-a-1, II-a-2, , II-b-1, II-b-2, II-c-1, II-c-2, II-d-1, II-d-2, or salt forms thereof, each of which is independently incorporated herein by reference; U.S. Pat. No. 9,394,333, U.S. Pat. No. 9,744,183, U.S. Pat. No. 9,605,019, U.S. Pat. No. 9,982,257, U.S. Patent Application Publication No. 20170037399, U.S. Patent Application Publication No. 20180216108, U.S. Patent Application Publication No. 20180216 107, U.S. Patent No. 9598458, WO 2017 / 062862, WO 2018 / 067973, WO 2017 / 160741, WO 2017 / 192679, WO 2017 / 210647, WO 2018 / 098264, WO 2018 / 022473, WO 2018 / 223056, WO 2018 / 223073, WO 2018 / 223 081, WO 2018 / 237194, WO 2019 / 032607, WO 2019032612, WO 2019 / 055951, WO 2019 / 075357, WO 2019 / 200185, WO 2019 / 217784, and / or WO 2019 / 032612. In a typical linkage, as in natural DNA and RNA, an internucleotide bond forms a bond with two sugars (which may be unmodified or modified as described herein).In many embodiments, as exemplified herein, the internucleotide linkages are formed by one optionally modified ribose or deoxyribose at the 5' carbon and another optionally modified ribose or deoxyribose at the 3' carbon via an oxygen atom or heteroatom (e.g., Y and Z in the various formulas). In certain embodiments, each nucleoside unit independently linked by an internucleotide linkage independently comprises a nucleobase that is independently optionally substituted A, T, C, G, or U, or a substituted tautomer of A, T, C, G, or U, or a nucleobase comprising an optionally substituted heterocyclyl and / or heteroaryl ring having at least one nitrogen atom.

[0341] In some embodiments, the bond is -YP L (-XR L )-Z- or a salt form thereof, wherein: P L is P, P(=W), P->B(-L L -R L )3, or P N and; W is O, N(-L L -R L ), S, or Se; P N , P=NC(-L L -R')(=L N -R'), or P=NL L -R L and; L N is =NL L1 -, =CH-L L1 - (wherein CH is optionally substituted), or =N + (R')(Q - )-L L1 - and; Q - is an anion; Each of X, Y and Z independently represents -O-, -S-, -L L -N(-L L -R L )-L L-, -L L -N=C(-L L -R L )-L L - or L L and; Each R L independently, -L L -N(R')2, -L L -R', -N=C(-L L -R')2, -L L -N(R')C(NR')N(R')2, -L L -N(R')C(O)N(R')2, a hydrocarbon, or one or more additional chemical moieties optionally linked via a linker; L L1 and L L each of which is independently L; -Cy IL - is -Cy-; Each L is independently a covalent bond or C 1~30 C with an aliphatic group and 1 to 10 heteroatoms 1~30 heteroaliphatic groups, wherein one or more methylene units are optionally and independently selected from C 1~6 Alkylene, C 1~6Alkenylene, -C≡C-, divalent C1-C6 heteroaliphatic group having 1 to 5 heteroatoms, -C(R')2-, -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(NR')N(R')-, -N(R')C(NR')N(R')-, -C(O)N(R' )-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, -C (O)O-, -P(O)(OR')-, -P(O)(SR')-, -P(O)(R')-, -P(O)(NR')-, -P(S)(OR')-, -P(S)(S and -R')-, -P(S)(R')-, -P(S)(NR')-, -P(R')-, -P(OR')-, -P(SR')-, -P(NR')-, -P(OR')[B(R')3]-, -OP(O)(OR')O-, -OP(O)(SR')O-, -OP(O)(R')O-, -OP(O)(NR')O-, -OP(OR')O-, -OP(SR')O-, -OP(NR')O-, -OP(R')O-, -OP(OR')[B(R')3]O-, and -[C(R')2C(R')2O]n-, wherein n is 1 to 50, and one or more nitrogen or carbon atoms are optionally and independently replaced by Cy L Replaced by; each -Cy- is independently an optionally substituted divalent 3-30 membered monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms; Each Cy L is independently an optionally substituted trivalent or tetravalent 3-30 membered monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms; each R' is independently -R, -C(O)R, -C(O)N(R), -C(O)OR, or -S(O)R; Each R is independently -H or C 1~30 Aliphatic, C with 1-10 heteroatoms 1~30 Heteroaliphatic, C 6~30 Aryl, C 6~30 Arylaliphatic, C with 1-10 heteroatoms 6~30an optionally substituted group selected from arylheteroaliphatic, 5-30 membered heteroaryl having 1-10 heteroatoms, and 3-30 membered heterocyclyl having 1-10 heteroatoms; or Two R groups, optionally and independently, together form a covalent bond, or: two or more R groups on the same atom optionally and independently, taken together with that atom, form an optionally substituted 3-30 membered monocyclic, bicyclic, or polycyclic ring having, in addition to that atom, 0-10 heteroatoms; or Two or more R groups on two or more atoms may optionally and independently be taken together with their intervening atoms to form an optionally substituted 3-30 membered monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms in addition to the intervening atoms.

[0342] In some embodiments, the internucleotide linkage is -OP L (-XR L )-O-, where each variable is independently as described herein. In some embodiments, the internucleotide linkage is -OP(=W)(-XR L )-O-, where each variable is independently as described herein. In some embodiments, the internucleotide linkage is -OP(=W)[-N(-L L -R L )-R L ]-O-, where each variable is independently as described herein. In some embodiments, the internucleotide linkage is -OP(=W)(-NH-L L -R LIn some embodiments, the internucleotide linkage has the structure -OP(=W)[-N(R')2]-O-, where each variable is independently as described herein. In some embodiments, the internucleotide linkage has the structure -OP(=W)(-NHR')-O-, where each variable is independently as described herein. In some embodiments, the internucleotide linkage has the structure -OP(=W)(-NHS02R)-O-, where each variable is independently as described herein. In some embodiments, the internucleotide linkage has the structure -OP(=W)[-N=C(-L L In some embodiments, an internucleotide linkage has the structure -OP(=W)[-N=C[N(R')2]2]-O-, where each variable is independently as described herein. In some embodiments, an internucleotide linkage has the structure -OP(=W)(-N=C(R")2)-O-, where each variable is independently as described herein. In some embodiments, an internucleotide linkage has the structure -OP(=W)(-N=C(R")2)-O-, where each variable is independently as described herein. In some embodiments, an internucleotide linkage has the structure -OP(=W)(-N(R")2)-O-, where each variable is independently as described herein. In some embodiments, W is O. In some embodiments, W is S. In some embodiments, such internucleotide linkages are non-negatively charged internucleotide linkages. In some embodiments, such internucleotide linkages are neutral internucleotide linkages.

[0343] In some embodiments, the internucleotide linkage is -P L (-XR L In some embodiments, the internucleotide linkage has the structure -P L (-XR LIn some embodiments, the internucleotide linkage has the structure -P(=W)(-XR L )-O-, where each variable is independently as described herein. In some embodiments, the internucleotide linkage is -P(=W)[-N(-L L -R L )-R L ]-O-, where each variable is independently as described herein. In some embodiments, the internucleotide linkage is -P(=W)(-NH-L L -R L In some embodiments, the internucleotide linkage has the structure -P(=W)[-N(R')2]-O-, where each variable is independently as described herein. In some embodiments, the internucleotide linkage has the structure -P(=W)(-NHR')-O-, where each variable is independently as described herein. In some embodiments, the internucleotide linkage has the structure -P(=W)(-NHS02R)-O-, where each variable is independently as described herein. In some embodiments, the internucleotide linkage has the structure -P(=W)[-N=C(-L LIn some embodiments, an internucleotide linkage has the structure -P(=W)[-N=C[N(R')2]2]-O-, where each variable is independently as described herein. In some embodiments, an internucleotide linkage has the structure -P(=W)(-N=C(R")2)-O-, where each variable is independently as described herein. In some embodiments, an internucleotide linkage has the structure -P(=W)(-N=C(R")2)-O-, where each variable is independently as described herein. In some embodiments, an internucleotide linkage has the structure -P(=W)(-N(R")2)-O-, where each variable is independently as described herein. In some embodiments, W is O. In some embodiments, W is S. In some embodiments, such internucleotide linkages are non-negatively charged internucleotide linkages. In some embodiments, such internucleotide linkages are neutral internucleotide linkages. In some embodiments, the P of such internucleotide linkages is attached to the N of the sugar.

[0344] In some embodiments, the linkage is a phosphorylguanidine internucleotide linkage. In some embodiments, the linkage is a thio-phosphorylguanidine internucleotide linkage.

[0345] In some embodiments, one or more methylene units are optionally and independently replaced with a moiety as described herein. L is or includes -SO2-. In some embodiments, L or L L is or includes -SON(R')-. In some embodiments, L or L L is or includes -C(O)-. In some embodiments, L or L L is or includes -C(O)O-. In some embodiments, L or L L is or includes -C(O)N(R')-. In some embodiments, L or L Lis or includes -P(=W)(R')-. In some embodiments, L or L L is or includes -P(=O)(R')-. In some embodiments, L or L L is or includes -P(=S)(R')-. In some embodiments, L or L L is or includes -P(R')-. In some embodiments, L or L L is or includes -P(=W)(OR')-. In some embodiments, L or L L is or includes -P(=O)(OR')-. In some embodiments, L or L L is or includes -P(=S)(OR')-. In some embodiments, L or L L is or contains -P(OR')-.

[0346] In some embodiments, -XR L is -N(R')SO2R L In some embodiments, -XR L is -N(R')C(O)R L In some embodiments, -XR L is -N(R')P(=O)(R')R L is.

[0347] In some embodiments, the linkage, e.g., a non-negatively charged internucleotide linkage or a neutral internucleotide linkage, is -P(=W)(-N=C(R")2)-, -P(=W)(-N(R')S02R")-, -P(=W)(-N(R')C(O)R")-, -P(=W)(-N(R")2)-, -P(=W)(-N(R')P(O)(R")2)-, -OP(=W)(-N=C(R")2)O-, -OP(=W)(-N(R')S02R")O-, -OP(= having or comprising the structure of -OP(=W)(-N(R')C(O)R")O-, -OP(=W)(-N(R')P(O)(R")O-, -P(=W)(-N=C(R")O-, -P(=W)(-N(R')SOR")O-, -P(=W)(-N(R')C(O)R")O-, -P(=W)(-N(R")O- or -P(=W)(-N(R')P(O)(R")O-, or a salt thereof, wherein W is O or S; each R″ is independently R′, —OR′, —P(═W)(R′)2, or —N(R′)2; each R' is independently -R, -C(O)R, -C(O)N(R), -C(O)OR, or -S(O)R; Each R is independently -H or C 1~30 Aliphatic, C with 1-10 heteroatoms 1~30 Heteroaliphatic, C 6~30 Aryl, C 6~30 Arylaliphatic, C with 1-10 heteroatoms 6~30 an optionally substituted group selected from arylheteroaliphatic, 5-30 membered heteroaryl having 1-10 heteroatoms, and 3-30 membered heterocyclyl having 1-10 heteroa...

Claims

1. A double-stranded RNAi (dsRNAi) agent comprising a guide strand and a passenger strand, a) the guide strand is complementary or substantially complementary to an HSD17B13 target RNA sequence; and i. backbone phosphorothioate chiral centers in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide; ii. Rp, Sp or alternating backbone phosphorothioate chiral centers between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide; iii. one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration upstream of a backbone phosphorothioate chiral center in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide; and / or iv. one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide, and between the (+2) nucleotide and the immediately downstream (+3) nucleotide, and between (a) the (+3) nucleotide and the (+4) nucleotide, and (b) the (+5) nucleotide and the (+6) nucleotide; Including; b) the guide strand comprises one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds between any two adjacent nucleotides between the second (+2) nucleotide relative to the 5'-terminal nucleotide of the guide strand and the 3'(N-1) nucleotide immediately preceding the end of the guide strand, where N is the 3'-terminal nucleotide; c) the guide strand comprises a 2' modification of the 3' nucleotide of a nucleotide pair linked by an Rp, Sp, or stereorandom non-negatively charged internucleotide bond; d) the passenger strand is i. 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, where n is about 1 to 49; and ii. one or more backbone chiral centers of Rp or Sp configuration and e) each strand of the dsRNAi agent independently has a length of from about 15 to about 49 nucleotides; f) the dsRNAi is capable of inducing HSD17B13-specific RNA interference; Double-stranded RNAi (dsRNAi) agents.

2. 1. A chiral control oligonucleotide composition comprising a double-stranded oligonucleotide, wherein the guide strand and passenger strand of the double-stranded oligonucleotide independently comprise: a) common base sequence and length; b) a common backbone bonding pattern; and c) Pattern of common skeletal chiral centers characterized by; chiral controlled in that oligonucleotides having a common pattern of chiral centers are enriched for substantially racemic preparations of guide strands having the same common base sequence and length; a) the guide strand is complementary or substantially complementary to an HSD17B13 target RNA sequence; and i. backbone phosphorothioate chiral centers in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide; ii. Rp, Sp or alternating backbone phosphorothioate chiral centers between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide; iii. one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration upstream of a backbone phosphorothioate chiral center in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide; and / or iv. one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide, and between the (+2) nucleotide and the immediately downstream (+3) nucleotide, and between (a) the (+3) nucleotide and the (+4) nucleotide, and (b) the (+5) nucleotide and the (+6) nucleotide; or b) the guide strand comprises one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds between any two adjacent nucleotides between the second (+2) nucleotide relative to the 5'-terminal nucleotide of the guide strand and the 3'(N-1) nucleotide immediately preceding the end of the guide strand, where N is the 3'-terminal nucleotide; c) the guide strand comprises a 2' modification of the 3' nucleotide of a nucleotide pair linked by an Rp, Sp, or stereorandom non-negatively charged internucleotide bond; d) the passenger strand is i. 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, where n is about 1 to 49; and ii. one or more backbone chiral centers of Rp or Sp configuration and e) the guide and passenger strands have a length of about 15 to about 49 nucleotides; and f) the guide and passenger strands are capable of inducing HSD17B13-specific RNA interference; composition.

3. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the guide strand comprises a backbone phosphorothioate chiral center in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, where n is about 1 to 49.

4. 3. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the guide strand comprises Rp, Sp, or alternating backbone phosphorothioate chiral centers between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, where n is about 1 to 49.

5. 3. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the guide strand comprises one or more backbone phosphorothioate chiral centers of the Rp or Sp configuration upstream of backbone phosphorothioate chiral centers of the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, where n is about 1 to 49.

6. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the guide strand comprises one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds between any two adjacent nucleotides between the second (+2) nucleotide relative to the 5'-terminal nucleotide of the guide strand and the 3'(N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide), and the passenger strand comprises 0 to n non-negatively charged internucleotide bonds (where n is about 1 to 49).

7. 3. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the guide strand comprises a backbone phosphorothioate chiral center in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide, and the passenger strand comprises one or more backbone chiral centers in the Rp or Sp configuration.

8. 3. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the guide strand comprises a backbone phosphorothioate chiral center in an Rp, Sp or alternating configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the +2 nucleotide and the immediately downstream (+3) nucleotide, and the passenger strand comprises one or more backbone chiral centers in an Rp or Sp configuration.

9. 3. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the guide strand comprises one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration upstream of a backbone phosphorothioate chiral center in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide, and the passenger strand comprises one or more backbone chiral centers in the Rp or Sp configuration.

10. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the guide strand comprises one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide, between the (+2) nucleotide and the immediately downstream (+3) nucleotide, and between (a) the (+3) nucleotide and the (+4) nucleotide, and (b) the (+5) nucleotide and the (+6) nucleotide.

11. The guide strand is 【Chemistry 1】 (Bases: A, C, G, T, U, abasic and modified nucleobases, R: H, OH, O-alkyl, F, MOE, LNA bridge at 4' position, BNA bridge at 4' position) The double-stranded oligonucleotide of claim 1 or the composition of claim 2, comprising a 5'-end modification selected from:

12. 3. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the guide strand comprises one or more Rp, Sp, or stereorandom non-negatively charged internucleotide bonds between any two adjacent nucleotides between the second (+2) nucleotide relative to the 5'-terminal nucleotide of the guide strand and the 3'(N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide), and the passenger strand comprises one or more backbone chiral centers in the Rp or Sp configuration.

13. 3. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the guide strand comprises a backbone phosphorothioate chiral center in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, where n is about 1 to 49, and one or more backbone chiral centers in the Rp or Sp configuration.

14. 3. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the guide strand comprises an Rp, Sp, or alternating backbone phosphorothioate chiral center between the 5'-terminal (+1) nucleotide and the immediately downstream (+2) nucleotide and between the (+2) nucleotide and the immediately downstream (+3) nucleotide, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, where n is about 1 to 49, and one or more backbone chiral centers in the Rp or Sp configuration.

15. 3. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the guide strand comprises one or more backbone phosphorothioate chiral centers in the Rp or Sp configuration upstream of backbone phosphorothioate chiral centers in the Sp configuration between the 3'-terminal nucleotide and the pre-terminal (N-1) nucleotide and between the pre-terminal (N-1) nucleotide and the immediately upstream (N-2) nucleotide, and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages, where n is about 1 to 49, and one or more backbone chiral centers in the Rp or Sp configuration.

16. 3. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the guide strand comprises one or more Rp, Sp, or stereorandom non-negatively charged internucleotide linkages between any two adjacent nucleotides between the second (+2) nucleotide relative to the 5'-terminal nucleotide of the guide strand and the 3' (N-1) nucleotide immediately preceding the end of the guide strand (where N is the 3'-terminal nucleotide), and the passenger strand comprises 0 to n Rp, Sp, or stereorandom non-negatively charged internucleotide linkages (where n is about 1 to 49) and one or more backbone chiral centers in the Rp or Sp configuration.

17. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the Rp, Sp or stereorandom non-negatively charged internucleotide bond has a neutral charge.

18. The neutral backbone internucleotide bond is 【Chemistry 2】 18. The double-stranded oligonucleotide or composition of claim 17, wherein:

19. The guide strand may have a nucleotide sequence having the following structure between the third (+3) and fourth (+4) nucleotides of the guide strand, between the tenth (+10) and eleventh (+11) nucleotides of the guide strand, or both: 【Transformation 3】 20. The double-stranded oligonucleotide or composition of claim 18, comprising a linkage having the formula:

20. The passenger strand may have the following structure at the 5' position of the central nucleotide of the passenger strand, the 3' position of the central nucleotide of the passenger strand, or both: 【Chemistry 4】 20. The double-stranded oligonucleotide or composition of claim 19, comprising a linkage having the formula:

21. 3. The composition of claim 2, wherein the guide strands and passenger strands in the composition independently share a common base sequence, a common base modification pattern, a common sugar modification pattern, and / or a common internucleotide linkage pattern, and the guide strands and passenger strands in the composition are at least 90% of all the guide strands and passenger strands in the composition.

22. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the double-stranded oligonucleotide comprises a carbohydrate moiety linked at an internucleoside internucleotide bond, optionally via a linker.

23. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the double-stranded oligonucleotide comprises a lipid moiety linked to the double-stranded oligonucleotide at an internucleoside, internucleotide bond, optionally via a linker.

24. 3. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein one or both strands of the double-stranded oligonucleotide comprise targeting moieties linked in internucleoside, internucleotide linkages, optionally via a linker.

25. 3. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of the internucleotide linkages of the double-stranded oligonucleotide are independently chiral internucleotide linkages.

26. 10. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein at least 3%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% of the nucleotide units of the double-stranded oligonucleotide independently comprise a 2'-substitution.

27. 27. The double-stranded oligonucleotide or composition of claim 26, wherein the 2'-substitution of the oligonucleotide is 2'-F.

28. 27. The double-stranded oligonucleotide or composition of claim 26, wherein the 2'-substitution of the oligonucleotide is 2'-OR1.

29. 27. The double-stranded oligonucleotide or composition of claim 26, wherein the 2'-substitution of the oligonucleotide is -L-, where L links C2 and C4 of the sugar unit.

30. 10. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein at least 3%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97% of the nucleotide units of the double-stranded oligonucleotide independently comprise a 2'-substitution.

31. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the guide strand comprises an HSD17B13 target binding sequence that is fully complementary to an HSD17B13 target sequence, the HSD17B13 target binding sequence having a length of at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 bases, each base being an optionally substituted adenine, cytosine, guanosine, thymine or uracil, and the HSD17B13 target sequence comprises one or more allelic sites (wherein the allelic site is a SNP or a mutation).

32. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the HSD17B13 target sequence comprises two SNPs.

33. The double-stranded oligonucleotide of claim 1 or the composition of claim 2, wherein the HSD17B13 target sequence comprises an allele site, and the HSD17B13 target binding sequence is fully complementary to the HSD17B13 target sequence of a disease-associated allele, but is not complementary to the sequence of an allele less associated with the disease.

34. the double-stranded oligonucleotide comprises a guide strand that binds to a transcription product of an HSD17B13 target nucleic acid sequence, wherein a plurality of alleles exist in the population, each allele containing a specific nucleotide signature sequence element that defines the allele relative to other alleles of the same HSD17B13 target nucleic acid sequence; the base sequence of the guide strand is complementary to or includes the characteristic sequence element that defines the allele; the guide strand, when contacted with a cell containing a transcript of the HSD17B13 target nucleic acid sequence, exhibits repression of the transcript of the allele or the protein encoded thereby at a level greater than the level of repression observed for another allele of the same nucleic acid sequence; The double-stranded oligonucleotide of claim 1 or the composition of claim 2.

35. a guide strand comprising WV-47139 and a passenger strand comprising WV-42589; a guide strand comprising WV-47159 and a passenger strand comprising WV-42589; a guide strand comprising WV-49590 and a passenger strand comprising WV-42589; or A guide strand containing WV-49591 and a passenger strand containing WV-42589 The double-stranded oligonucleotide of claim 1 or the composition of claim 2, comprising:

36. A method for reducing the level and / or activity of an HSD17B13 transcript or the protein encoded thereby, comprising administering to a cell expressing the HSD17B13 transcript a double-stranded oligonucleotide described in claim 1, wherein the guide strand of the double-stranded oligonucleotide or composition comprises an HSD17B13 binding sequence that is fully complementary to the HSD17B13 target sequence of the transcript.

37. 37. The method of claim 36, wherein the cell is an immune cell, a blood cell, a heart cell, a lung cell, a photoreceptor cell, a muscle cell, a liver cell, a kidney cell, a brain cell, a central nervous system cell, or a peripheral nervous system cell.

38. 1. A method for allele-specifically silencing HSD17B13 transcripts from a nucleic acid sequence, wherein a plurality of alleles exist within a population, each allele containing a specific nucleotide signature sequence element that defines the allele relative to other alleles of the same HSD17B13 target nucleic acid sequence, comprising: contacting a sample containing a transcript of the HSD17B13 target nucleic acid sequence with the double-stranded oligonucleotide of claim 1; the guide strand of the double-stranded oligonucleotide or composition comprises an HSD17B13 binding sequence that is identical to or completely complementary to an HSD17B13 target sequence in the nucleic acid sequence, the HSD17B13 binding sequence comprising a characteristic sequence element that defines a target allele; When the guide strand of the double-stranded oligonucleotide or composition is contacted with a cell containing transcripts of both the target allele and another allele of the same nucleic acid sequence, the transcript of the target allele is suppressed at a level that exceeds the level of suppression observed for the other allele of the same nucleic acid sequence. method.

39. 1. A method for allele-specifically silencing HSD17B13 transcripts from a nucleic acid sequence, wherein a plurality of alleles exist within a population, each allele containing a specific nucleotide signature sequence element that defines the allele relative to other alleles of the same HSD17B13 target nucleic acid sequence, comprising: Administering to a subject comprising a transcript of the HSD17B13 target nucleic acid sequence the double-stranded oligonucleotide of claim 1; the guide strand of the double-stranded oligonucleotide or composition comprises an HSD17B13 binding sequence that is identical to or completely complementary to an HSD17B13 target sequence in the nucleic acid sequence, the HSD17B13 binding sequence comprising a characteristic sequence element that defines a target allele; When the guide strand of the double-stranded oligonucleotide or composition is contacted with a cell containing transcripts of both the HSD17B13 target allele and another allele of the same nucleic acid sequence, the transcript of the target allele is suppressed at a level greater than the level of suppression observed for the other allele of the same nucleic acid sequence. method.

40. When the oligonucleotide is contacted with a cell containing transcripts of both the HSD17B13 target allele and another allele of the same nucleic acid sequence, a) a level greater than that in the absence of said composition; b) a level of suppression that exceeds the level of suppression observed for another allele of the same nucleic acid sequence; or c) a level that is greater than that observed in the absence of the composition and that is greater than the level of suppression observed for another allele of the same nucleic acid sequence 40. The method of any one of claims 36 to 39, wherein the method exhibits suppression of the transcript of the target allele.

41. 41. The method of claim 40, wherein the cell is an immune cell, a blood cell, a heart cell, a lung cell, a photoreceptor cell, a muscle cell, a liver cell, a kidney cell, a brain cell, a central nervous system cell, or a peripheral nervous system cell.

42. 40. The method of any one of claims 36 to 39, wherein the suppression of HSD17B13 transcript of the target allele is greater than in the absence of the composition and at a level that exceeds the level of suppression observed for another allele of the same nucleic acid sequence.

43. A non-human animal that has been engineered to contain an HSD17B13 polypeptide or a characteristic portion thereof.

44. A non-human animal that has been engineered to contain and / or express a polynucleotide whose sequence encodes a HSD17B13 polypeptide or a characteristic portion thereof.

45. 45. The animal of claim 43 or 44, wherein the genome of said animal comprises a polynucleotide whose sequence encodes a HSD17B13 polypeptide or a characteristic portion thereof.

46. 45. The animal of claim 43 or 44, wherein the germline genome of said animal comprises a polynucleotide whose sequence encodes a HSD17B13 polypeptide or a characteristic portion thereof.

47. 45. The animal of claim 43 or 44, wherein the animal is a rodent.

48. 48. The rodent of claim 47, wherein the animal is a mouse.

49. 48. The rodent of claim 47, wherein the rodent is a rat.

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