Patterned modified oligonucleotides for extended duration of action

Abstract

Provided are oligomeric agents having increased metabolic stability, wherein the agent can modulate the amount or activity of a target RNA in a cell or a subject, and in certain instances modulate the amount of a protein encoded by such RNA in a cell or a subject. Also provided are methods of using such oligomeric agents, and pharmaceutical compositions thereof. Such oligomeric agents, methods, and pharmaceutical compositions are useful in the treatment of one or more diseases or conditions.

Description

[0001] CHEMO 113W0

[0002] PATTERNED MODIFIED OLIGONUCLEOTIDES FOR EXTENDED DURATION OF ACTION

[0003] Sequence Listing

[0004] The present application is being filed concurrently with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled CHEM0113SEQ.xml created on August 14, 2025, which is 15 KB in size. The contents of the electronic format of the sequence listing are incorporated herein by reference in their entirety.

[0005] Field

[0006] Provided herein are oligomeric agents and compositions thereof, where the oligomeric agents comprise modified oligonucleotides having nucleosides in particular patterns. Also provided are methods of using such agents in the treatment of a disease or condition.

[0007] Background

[0008] Antisense compounds comprising a variety of chemical modifications and motifs have been reported. In certain instances antisense compounds have been shown to modulate protein expression by binding to a target messenger RNA (mRNA) or pre-mRNA encoding the protein, which may result in cleavage (“knockdown”) of the mRNA. However, such compounds require chemical modification in order to have acceptable properties of distribution, longevity, and selectivity in vivo. New chemical modifications allow incredible variability, and implementing certain patterns of modifications may provide superior properties for a given nucleobase sequence.

[0009] There remains a need for new antisense compounds having improved properties for use in pharmaceutical products.

[0010] Summary

[0011] The present disclosure provides an oligomeric agent comprising a modified oligonucleotide for modulating expression of a target gene. Such oligonucleotides generally comprise a 5 ’-adenosine having a modified sugar moiety, where the sugar moiety is a bicyclic sugar moiety such as a constrained ethyl (cEt) or a locked nucleic acid (LNA). The oligonucleotides also comprise a deoxy region including contiguous deoxynucleotides flanked by a 3 ’-wing and a 5 ’-wing comprising sugar-modified nucleosides. In certain embodiments, oligomeric agents comprise a pattern of modified oligonucleotides, in particular those comprising sugar-modified nucleosides in the wing of a gapmer and / or an extended gap region. In certain embodiments, the oligomeric agent is an oligomeric duplex. The oligomeric agent may be formulated as pharmaceutical composition for use in modulating the amount or activity of a target RNA. Modified oligonucleotides and the compositions comprising them, including, but not limited to, oligomeric agents, oligomeric duplexes, antisense agents, and pharmaceutical compositions described herein, are useful for modulating target expression in a cell. Such oligomeric agents may provide CHEMO 113W0 enhanced metabolic stability over analogous oligomeric agents having an adenosine in the 5 ’-most nucleoside position with a sugar moiety other than a bicyclic sugar moiety. In certain embodiments, provided are oligomeric agents comprising a 5 ’-terminal nucleoside comprising a cEt or LNA sugar moiety that exhibit enhanced metabolic stability compared to an analogous oligomeric agent including a sugar moiety other than a bicyclic sugar moiety. In certain embodiments, the bicyclic sugar moiety is cEt. In certain embodiments, the bicyclic sugar moiety is LNA.

[0012] Detailed Description

[0013] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive. Herein, the use of the singular includes the plural unless specifically stated otherwise. Furthermore, the use of the term “including” as well as other forms, such as “includes” and “included”, is not limiting. Also, terms such as “element” or “component” encompass both elements and components comprising one unit and elements and components that comprise more than one subunit, unless specifically stated otherwise.

[0014] DEFINITIONS

[0015] The following definitions are provided, along with additional definitions throughout the specification, for a complete understanding of the instant invention. Unless specific definitions are provided herein, nomenclature used in connection with, and procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well-known and commonly used in the art. Unless otherwise indicated, certain terms have the following meanings:

[0016] As used herein, a substituent at the “2 '-position” means that the substituent is directly attached to the carbon at the 2'-position of a furanosyl sugar moiety.

[0017] As used herein, “2'-deoxynucleoside” means a nucleoside comprising a 2'-deoxyfuranosyl sugar moiety (a 2’-H(H) furanosyl sugar moiety). A 2'-deoxynucleoside may be a 2'-P-D-deoxynucleoside which comprises a 2'-P-D-deoxyribosyl sugar moiety, and which is in the P-D ribosyl configuration as found in naturally occurring deoxyribonucleic acid (DNA). A 2'-deoxynucleoside or a nucleoside comprising an unmodified 2'-deoxyribosyl sugar moiety may be abasic, comprise a modified nucleobase, or may comprise an RNA nucleobase (uracil).

[0018] As used herein, “2'-deoxy sugar moiety” means a 2'-H(H) deoxyfiiranosyl sugar moiety.

[0019] As used herein, “2'-MOE” means a 2'-OCH2CH2OCH3 group at the 2'-position of a furanosyl sugar moiety. A “2'-MOE sugar moiety” means a sugar moiety with a 2'-OCH2CH2OCH3 group at the 2'-position of a furanosyl sugar moiety. Unless otherwise indicated, a 2'-MOE sugar moiety is in the P-D-ribosyl stereochemical configuration. “MOE” means O -methoxy ethyl.

[0020] As used herein, “2'-MOE nucleoside” or “2'-OCH2CH2OCH3 nucleoside” means a nucleoside comprising a 2'-MOE sugar moiety (or 2'-OCH2CH2OCH3 furanosyl sugar moiety). CHEMO 113W0

[0021] As used herein, “2'-0Me” means a 2'-0CHs group at the 2'-position of a furanosyl sugar moiety. A “2'-OMe sugar moiety” means a sugar moiety with a 2'-OCHs group at the 2'-position of a furanosyl sugar moiety. Unless otherwise indicated, a 2'-OMe sugar moiety is in the P-D-ribosyl stereochemical configuration.

[0022] As used herein, “2'-OMe nucleoside” means a nucleoside comprising a 2'-OMe sugar moiety.

[0023] As used herein, “2'-F” means a 2'-fluoro group at the 2'-position of a furanosyl sugar moiety. A “2'-F sugar moiety” means a sugar moiety with a 2'-F group at the 2'-position of a furanosyl sugar moiety. Unless otherwise indicated, a 2'-F sugar moiety is in the P-D-ribosyl configuration.

[0024] As used herein, “2'-F nucleoside” means a nucleoside comprising a 2'-F sugar moiety.

[0025] As used herein “2'-NMA” means a 2'-OCH2C(=O)-N(H)CH3group at the 2'-position of a furanosyl sugar moiety. A “2'-NMA sugar moiety” means a sugar moiety with a 2'-OCH2C(=O)-N(H)CH3group at the 2'-position of a furanosyl sugar moiety.

[0026] As used herein, “2'-NMA nucleoside” means a nucleoside comprising a 2'-NMA sugar moiety.

[0027] As used herein, “2'-substituted nucleoside” means a modified nucleoside comprising a 2'- substituted furanosyl sugar moiety.

[0028] As used herein, “2'-substituted sugar moiety” means a modified furanosyl sugar moiety wherein the 2'-position is attached to at least one substituent other than H or OH. A 2'-substituted sugar moiety includes a bicyclic sugar moiety wherein the second ring is joined to the furanosyl ring at the 2'-position. 2'-substituted sugar moieties include, but are not limited to, 2'-OMe sugar moieties, 2'-MOE sugar moieties, 2'-F sugar moieties, 2'-NMA sugar moieties, cEt sugar moieties, and LNA sugar moieties.

[0029] As used herein, “stop site” refers to the 3 '-most nucleotide of a target nucleic acid which is complementary to an oligonucleotide when the oligonucleotide is hybridized to the target nucleic acid.

[0030] As used herein, “start site” refers to the 5 '-most nucleotide of a target nucleic acid which is complementary to an oligonucleotide when the oligonucleotide is hybridized to the target nucleic acid.

[0031] As used herein, “5 -methylcytosine” means a cytosine modified with a methyl group attached at the 5 position. A 5 -methylcytosine is a modified nucleobase.

[0032] As used herein, “abasic nucleoside” means a modified nucleoside in which the nucleobase is absent.

[0033] As used herein, “ameliorate” with reference to a symptom of a disease, means improvement in, or lessening of, or preclusion of, at least one symptom of a disease. As used herein, “disease” includes disorders, conditions, and injuries. Amelioration may be reduction in severity or frequency of a symptom or the delayed onset, prevention of occurrence of, or slowing of progression in the severity or frequency of, a symptom. Progression, frequency, or severity indicators may be determined by subjective or objective measures known in the art and / or described herein.

[0034] As used herein, “antisense activity” means any detectable and / or measurable change attributable (whether directly and / or indirectly) to hybridization of an antisense oligonucleotide to a target nucleic acid. For example, compounds have antisense activity when they alter the amount or activity of a target nucleic acid by 25% or more in an in vitro assay; or, for example compounds have antisense activity when they CHEMO 113W0 alter the amount or activity of a target nucleic acid by 25% or more in an in vivo assay. Antisense activity may be assessed in a standard assay. Herein, antisense activity is a modulation in the amount or expression of a target nucleic acid or a protein encoded by such target nucleic acid compared to target nucleic acid levels or target protein levels in the absence of the oligonucleotide.

[0035] As used herein, “antisense agent” means an oligomeric agent comprising an antisense oligonucleotide.

[0036] As used herein, “antisense oligonucleotide” means an oligonucleotide having at least one region (a “targeting region”) having a nucleobase sequence that is complementary to a target nucleic acid (e.g., a target region). An antisense oligonucleotide may be paired with a second oligonucleotide (herein, a “sense oligonucleotide”) having a nucleobase sequence that is complementary to the antisense oligonucleotide (for example, forming an “oligomeric duplex”), may be an unpaired antisense oligonucleotide (herein, a singlestranded antisense oligonucleotide), or may be a “hairpin oligonucleotide” that has at least one region that is self-complementary.

[0037] As used herein, “bicyclic sugar” or “bicyclic sugar moiety” means a modified sugar moiety comprising a furanosyl sugar moiety and a second ring, wherein the second ring is formed via a bridge connecting two non-geminal atoms in the ring of the furanosyl sugar moiety, thereby forming a bicyclic structure. Examples of bicyclic sugar moieties include locked nucleic acid (LNA) sugar moieties and constrained ethyl (cEt) sugar moieties as defined herein.

[0038] As used herein, “bicyclic nucleoside” or “BNA” means a nucleoside comprising a bicyclic sugar moiety. As used herein, “bicyclic adenosine” means a modified nucleoside comprising an adenine nucleobase and a bicyclic sugar moiety.

[0039] As used herein, “blunt” or “blunt ended” in reference to an oligomeric duplex means that there are no terminal unpaired nucleosides (z.e., no overhanging nucleosides). One or both ends of an oligomeric duplex may be blunt.

[0040] As used herein, “cell-targeting moiety” means a conjugate moiety or portion of a conjugate moiety that has affinity for a particular cell type or particular cell types. For example, a cell -targeting moiety may have affinity for a cell surface moiety, such as a cell surface receptor.

[0041] As used herein, “cerebrospinal fluid” or “CSF” means the fluid filling the space around the brain and spinal cord. “Artificial cerebrospinal fluid” or “aCSF” means a prepared or manufactured fluid that has certain properties (e.g., osmolarity, pH, and / or electrolytes) similar to cerebrospinal fluid and is biocompatible with CSF.

[0042] As used herein, “complementary nucleobase(s)” or “complementary” in reference to nucleobase(s) means nucleobases that form hydrogen bonds with one another . Complementary nucleobase pairs include, but are not limited to, adenine (A) and thymine (T), adenine (A) and uracil (U), cytosine (C) and guanine (G), 5 -methylcytosine (mC) and guanine (G). Certain modified nucleobases that are complementary to unmodified nucleobases or to other modified nucleobases are known in the art. For example, hypoxanthine, the nucleobase of the nucleoside inosine (I), can pair with adenine, cytosine, CHEMO 113W0 thymine, or uracil. Herein, hypoxanthine (I) is considered a complementary nucleobase to thymine (T), adenine (A), uracil (U), and cytosine (C).

[0043] As used herein, “complementary nucleobase sequence(s)” or “complementary” in reference to nucleobase sequence(s) refers to two nucleobase sequences in which some, a majority, or all of the nucleobases in the two nucleobase sequences are complementary nucleobases when the sequences are aligned. A “nucleobase sequence” means the order of contiguous nucleobases in a strand of linked nucleosides or a region thereof (e.g. , an oligonucleotide or region thereof, or a target nucleic acid or region thereof) independent of any sugar or intemucleoside linkage modification. Complementary nucleobase sequences may be nucleobase sequences of two separate strands of linked nucleosides or regions thereof (e.g. , an oligonucleotide and a region of a target nucleic acid, or an antisense oligonucleotide and its paired sense oligonucleotide) or complementary nucleobase sequences may be nucleobase sequences of two regions of a single strand of linked nucleosides (e.g., self-complementary regions of a hairpin oligonucleotide). As used herein, when a first strand of linked nucleosides (e.g., an oligonucleotide) or region thereof is described as being complementary to a second strand of linked nucleosides or region thereof (e.g. , a target nucleic acid or another oligonucleotide), it means that the nucleobase sequence of the first strand of linked nucleosides or region thereof is complementary to the nucleobase sequence of the second strand of linked nucleosides or region thereof when aligned. Not every pair of nucleobases in the aligned nucleobase sequences needs to be complementary for the two sequences to be “complementary.” Rather, some mismatches are tolerated. Where nucleobase sequence complementarity is expressed as a percent, such percent represents the percentage of nucleobases within one nucleobase sequence that are complementary to nucleobases within an equal length second nucleobase sequence when the nucleobase sequences are aligned. Unless otherwise specified, “complementary” is assumed to be at least 70%. Complementary nucleobase sequences may be 75%, 80%, 85%, 90%, 95%, or 100% complementary. For example, if a nucleobase sequence of an oligonucleotide consisting of 20 nucleosides is 80% complementary to another nucleobase sequence, then 16 of the nucleobase pairs are complementary nucleobases, and there are 4 mismatches when the sequences are aligned. If a nucleobase sequence of an oligonucleotide consisting of 20 nucleosides is at least 80% complementary to another nucleobase sequence, then 16, 17, 18, 19, or 20 of the nucleobase pairs are complementary nucleobases, and there are 0-4 mismatches when the sequences are aligned. As used herein, “fully complementary” or “100% complementary” means that each nucleobase pair of the two nucleobase sequences is complementary when the equal length sequences are aligned.

[0044] As used herein, “conjugate group” means a group of atoms that is directly or indirectly attached to an oligonucleotide. A conjugate group comprises a conjugate moiety and a conjugate linker that attaches the conjugate moiety to the oligonucleotide.

[0045] As used herein, “conjugate linker” means a single bond or a group of atoms comprising at least one bond that connects a conjugate moiety to an oligonucleotide.

[0046] As used herein, “conjugate moiety” means a group of atoms that when covalently bound to a molecule (e.g., an oligonucleotide) modifies one or more properties of such molecule compared to the CHEMO 113W0 same molecule lacking the conjugate moiety, wherein such properties include, but are not limited to pharmacodynamics, pharmacokinetics, stability, binding, absorption, tissue distribution, cellular distribution, cellular uptake, charge, and clearance.

[0047] As used herein, “complementary region” in reference to a strand of linked nucleosides (e.g., an oligonucleotide or a target nucleic acid) is a region of the strand of linked nucleosides in which the nucleobase sequence of the region is complementary with the nucleobase sequence of an equal-length region of a separate strand of linked nucleosides (e.g., an oligonucleotide and a target nucleic acid, or an antisense oligonucleotide and a sense oligonucleotide), or the nucleobase sequence of an equal-length region within the strand of linked nucleosides (e.g., in a “hairpin oligonucleotide”). A complementary region of a strand of linked nucleosides may be a portion of a strand of linked nucleosides or may include the entire strand of linked nucleosides. A complementary region may include a mismatch, but the nucleobases of the terminal nucleosides of a complementary region are complementary to the nucleobases of the terminal nucleosides of the equal -length region of the separate strand of linked nucleosides or to the nucleobases of the terminal nucleosides of the equal-length region within the strand of linked nucleosides. A “targeting region” of an oligonucleotide, is a complementary region in which the nucleobase sequence of the region is complementary to the nucleobase sequence of a target region of a target nucleic acid. A targeting region of a strand of linked nucleosides may be a portion of a strand of linked nucleosides or may include the entire strand of linked nucleosides. A “duplexing region” is a complementary region of an oligonucleotide (e.g., an antisense or sense oligonucleotide) having a nucleobase sequence that is complementary to the nucleobase sequence of a second oligonucleotide or region thereof. A duplexing region may be a portion of a strand of linked nucleosides or may include the entire strand of linked nucleosides.

[0048] As used herein, “constrained ethyl” or “cEt” or “cEt sugar moiety” means a P-D ribosyl bicyclic sugar moiety wherein the second ring of the bicyclic sugar is formed via a bridge connecting the 4'-carbon and the 2'-carbon of the -D ribosyl sugar moiety, wherein the bridge has the formula 4'-CH(CH3)-O-2', and wherein the methyl group of the bridge is in the S configuration.

[0049] As used herein, “cEt nucleoside” means a nucleoside comprising a cEt sugar moiety. A “cEt adenosine” means a modified nucleoside comprising an adenine nucleobase and a cEt sugar moiety.

[0050] As used herein, “deoxy region” means a region of 5-20 contiguous nucleosides, wherein at least 70% of the nucleosides are 2’-P-D-deoxyribosyl nucleosides. A deoxy region supports RNase H activity.

[0051] As used herein, “DNA sugar moiety” means a 2'-H(H) furanosyl sugar moiety, including a P-D ribosyl sugar moiety as found in naturally occurring DNA and stereoisomers thereof.

[0052] As used herein, “double-stranded” refers to hybridized or bound complementary regions, including those between two separate strands of linked nucleosides (e.g., an antisense oligonucleotide and a sense oligonucleotide) and those within a single strand of linked nucleosides (e.g., a hairpin oligonucleotide). Hybridized complementary regions of two separate strands of linked nucleosides form a “duplex” of the separate strands. Hybridized complementary regions of a single strand of linked nucleosides (i.e., a first CHEMO 113W0 region of the strand of linked nucleosides and a second region of the strand of linked nucleosides) form a “hairpin”.

[0053] As used herein, “duplex” means a structure formed by two separate strands of linked nucleosides (e.g., two separate oligonucleotides) or regions thereof, at least a portion of which are complementary to and hybridize to each other. For clarity, herein a “hairpin oligonucleotide” is one strand of linked nucleosides that comprises a region that is double stranded and is not a duplex.

[0054] As used herein, “hybridize” or “hybridization” means the act or process of two complementary regions of linked nucleosides (e.g., oligonucleotides, nucleic acids) annealing together to form a doublestranded region. While not limited to a particular mechanism, the most common mechanism of hybridization involves hydrogen bonding, which may be Watson-Crick, Hoogsteen, or reversed Hoogsteen hydrogen bonding, between complementary nucleobases.

[0055] As used herein, “intemucleoside linkage” means the covalent linkage between immediately adjacent (e.g., contiguous) nucleosides in an oligonucleotide. As used herein, “unmodified intemucleoside linkage” means a phosphodiester intemucleoside linkage. As used herein, “modified intemucleoside linkage” means any intemucleoside linkage other than a phosphodiester intemucleoside linkage. A “phosphorothioate intemucleoside linkage” is a modified intemucleoside linkage in which one of the non-bridging oxygen atoms of a phosphodiester intemucleoside linkage is replaced with a sulfur atom. A “mesyl phosphoramidate intemucleoside linkage” is a modified intemucleoside linkage in which one of the non-bridging oxygen atoms of a phosphodiester intemucleoside linkage is replaced with NS(=O)2CH3. Unless otherwise indicated, and in the context of linked nucleosides each comprising a furanosyl sugar moiety, an intemucleoside linkage joins the 3 '-carbon of one furanosyl sugar moiety to the 5 '-carbon of the other furanosyl sugar moiety.

[0056] As used herein, “inverted nucleoside” means a nucleoside having a 3' to 3' and / or 5' to 5' intemucleoside linkage.

[0057] As used herein, “linked nucleosides” are nucleosides that are connected in a contiguous sequence (z.e., nucleosides immediately adjacent to one another, no additional nucleosides are present between those that are linked).

[0058] As used herein, “locked nucleic acid” or “LNA” or “LNA sugar moiety” means a P-D ribosyl bicyclic sugar moiety wherein the second ring of the bicyclic sugar is formed via a bridge connecting the 4'-carbon and the 2'-carbon of the -D ribosyl sugar moiety, wherein the bridge has the formula 4'-CH-O- 2'.

[0059] As used herein, “LNA nucleoside” means a nucleoside comprising an LNA sugar moiety. An “LNA adenosine” means a modified nucleoside comprising an adenine nucleobase and an LNA sugar moiety.

[0060] As used herein, “metabolic stability” is a measure of the rate of metabolism of an oligomeric agent in contact with a metabolizing medium, where the metabolic process is deamination of an adenine nucleobase. In certain embodiments, the metabolizing medium is a cell. In certain embodiments, the metabolizing medium is a cell in vivo in a subject. In certain embodiments, the metabolizing medium CHEMO 113W0 includes human adenosine deaminase 2 (ADA2) enzyme, for example, as described herein in Example 4. In certain embodiments, the oligomeric agent comprises a bicyclic adenosine at its 5 ’-most position and the analogous oligomeric agent is identical other than the sugar moiety of the 5 ’-most adenosine nucleoside. In certain embodiments, the oligomeric agent comprises a cEt adenosine at its 5 ’-most position and the analogous oligomeric agent is identical other than the sugar moiety of the 5 ’-most adenosine nucleoside.

[0061] As used herein, a “mismatch” between two aligned nucleic acid sequences (e.g., nucleic acid sequence of separate strands of linked nucleosides or separate regions of a strand of linked nucleosides) means that the two nucleobases at a specified position of the aligned nucleobase sequences are not complementary nucleobases.

[0062] As used herein, “modified nucleoside” means a compound or subunit comprising a sugar moiety and optionally a nucleobase, wherein the sugar moiety is modified, and wherein the nucleobase is optionally modified or is absent (i.e., abasic nucleoside); modified nucleosides also include abasic nucleosides and nucleosides linked other than 3’- / 5’-, regardless of sugar structure.

[0063] As used herein, “modified sugar moiety” means a sugar moiety other than a RNA sugar moiety, or a stereoisomer thereof, or a DNA sugar moiety, or a stereoisomer thereof. A modified sugar moiety is selected from a modified furanosyl sugar moiety and a sugar surrogate. A “modified furanosyl sugar moiety” is a modified sugar moiety comprising a furanose ring. A modified furanosyl sugar moiety comprises a nucleobase at the 1 ’-position, and optional intemucleoside linkages at the 3’- and 5’- positions.

[0064] As used herein, a “modified nucleobase” means a nucleobase other than unmodified A, T, C, U, or G capable of pairing with at least one unmodified nucleobase. A “5 -methylcytosine” is a modified nucleobase. Inosine (I) is a nucleoside comprising the modified nucleobase hypoxanthine.

[0065] As used herein, “motif’ means a pattern of independently unmodified and / or independently modified sugar moieties, nucleobases, and / or intemucleoside linkages in an oligonucleotide.

[0066] As used herein, “non-bicyclic modified sugar moiety” means a modified furanosyl sugar moiety comprising a modification, such as a substituent, that does not form a bridge between two atoms of the sugar to form a second ring.

[0067] As used herein, "nucleobase" means an unmodified nucleobase or a modified nucleobase.

[0068] As used herein, “nucleobase sequence” means the order of contiguous nucleobases in a strand of linked nucleosides independent of any sugar or intemucleoside linkage modification. As used herein, “the nucleobase sequence of’ a reference SEQ ID NO refers only to the nucleobase sequence provided in such SEQ ID NO and therefore, unless otherwise indicated, includes compounds wherein each sugar moiety and each intemucleoside linkage, independently, is modified or unmodified, irrespective of the presence or absence of modifications indicated in the referenced SEQ ID NO.

[0069] As used herein, “nucleoside” means an “unmodified nucleoside” or a “modified nucleoside”. CHEMO 113W0

[0070] As used herein, “nucleoside overhang” or “overhang” refers to unpaired nucleosides at either or both ends of an oligomeric duplex or at an end of a hairpin structure. The nucleosides of an overhang are not part of the “duplexing region” of either of the two strands of linked nucleosides.

[0071] As used herein, "oligomeric agent" means a compound or complex comprising or consisting of at least one modified oligonucleotide and optionally one or more additional associated features selected from: (a) one or more additional modified or unmodified oligonucleotides, each of which may be hybridized to or covalently linked to the at least one modified oligonucleotide and / or to each other; (b) one or more conjugate groups, which may be covalently attached directly or indirectly to any oligonucleotide of such oligomeric agent; and (c) one or more terminal groups. Herein, where two oligonucleotides are described as being covalently attached to one another, such attachment is other than through a direct intemucleoside linkage. Thus, a single, unbranched oligonucleotide comprising only direct intemucleoside linkages cannot be described as two separate covalently linked oligonucleotides.

[0072] As used herein, “oligonucleotide” means a strand of linked nucleosides, wherein each nucleoside and / or each intemucleoside linkage of the strand of linked nucleosides may be independently modified or unmodified. Unless otherwise indicated, oligonucleotides consist of 12-80 linked nucleosides. Unless otherwise indicated, no more than 10% of the nucleosides of an oligonucleotide are abasic nucleosides. As used herein, “modified oligonucleotide” means an oligonucleotide, wherein at least one nucleoside and / or at least one intemucleoside linkage is modified. As used herein, “unmodified oligonucleotide” means an oligonucleotide consisting of unmodified nucleosides linked by phosphodiester intemucleoside linkages. An oligonucleotide may be paired with a second oligonucleotide that is complementary to the oligonucleotide to form an oligomeric duplex, or it may be unpaired.

[0073] As used herein, “pharmaceutical composition” means a mixture of substances suitable for administration to a subject. For example, a pharmaceutical composition may comprise an oligomeric agent and a sterile aqueous solution. A pharmaceutical composition may show activity in certain cell lines.

[0074] As used herein, “pharmaceutically acceptable diluent” means an ingredient in a pharmaceutical composition suitable for use in administering to a subject. Typically, a “diluent” lacks pharmacological activity but is desirable in preparing a pharmaceutical composition.

[0075] As used herein, “pharmaceutically acceptable salts” means physiologically and pharmaceutically acceptable salts of compounds. Pharmaceutically acceptable salts retain the desired biological activity of the parent compound and do not impart undesired toxicological effects thereto.

[0076] As used herein, “prodrug” means a therapeutic agent in a first form outside the body that is converted to a second form within an animal or cells thereof. Typically, conversion of a prodrug within the animal is facilitated by the action of an enzyme (e.g., endogenous or viral enzyme) or chemical present in cells or tissues and / or by physiologic conditions. The first form of the prodrug may be less active than the second form. CHEMO 113W0

[0077] As used herein, “RNA nucleoside” means a nucleoside comprising an unmodified RNA sugar moiety. An RNA nucleoside may comprise a modified or unmodified nucleobase. An RNA nucleoside may comprise a thymine nucleobase or a modified nucleobase, or may be an abasic nucleoside.

[0078] As used herein, “RNA sugar moiety” means a 2'-0H(H) furanosyl sugar moiety, including a P-D ribosyl sugar moiety as found in naturally occurring RNA and stereoisomers thereof.

[0079] As used herein, “RNAi agent” means an antisense agent that acts, at least in part, through RISC or Ago2 to modulate a target nucleic acid and / or a protein encoded by a target nucleic acid. RNAi agents include, but are not limited to double-stranded siRNA, single -stranded RNAi (ssRNAi), and microRNA, including microRNA mimics. RNAi agents may comprise conjugate groups and / or terminal groups. In certain embodiments, an RNAi agent modulates the amount and / or activity of a target nucleic acid. The term RNAi agent excludes antisense agents that act through RNase H.

[0080] As used herein, “RNase H agent” means an antisense agent that acts, at least in part, through RNase H to modulate a target nucleic acid and / or protein encoded by a target nucleic acid. RNase H agents may be single-stranded or RNase H agents may be double-stranded. RNase H compounds may comprise conjugate groups and / or terminal groups. RNase H agents may modulate the amount and / or activity of a target nucleic acid. The term RNase H agent excludes antisense agents that act principally through RISC / Ago2.

[0081] As used herein, “single -stranded” in reference to a strand of linked nucleosides (e.g., an oligonucleotide) means that the strand of linked nucleosides is not part of a duplex or part of a doublestranded region. Single-stranded nucleic acids (e.g., single-stranded oligonucleotides) are capable of hybridizing to complementary nucleic acids to form duplexes, at which point they are no longer singlestranded.

[0082] As used herein, “splice-modulating agent” means an antisense agent that acts, at least in part, by modulating the splicing of a target nucleic acid.

[0083] As used herein, “steric -blocking agent” means an antisense agent that acts, at least in part, due to directly binding to a target nucleic acid, thus blocking the interaction of the target nucleic acid with other nucleic acids or proteins.

[0084] As used herein, “stabilized phosphate moiety” means a 5 '-phosphate analog that is metabolically more stable than a 5 '-phosphate as naturally occurs on DNA or RNA.

[0085] As used herein, “stereorandom” or “stereorandom chiral center” in the context of a population of molecules of identical molecular formula means a chiral center that is not intentionally controlled during synthesis, or enriched following synthesis, for a particular absolute stereochemical configuration at that chiral center. It is understood that a stereorandom chiral center may not be racemic because one absolute configuration predominates following synthesis, e.g., due to steric and electronic interactions of reagents with the reactant molecule. The stereorandom chiral center may be at the phosphorous atom of a stereorandom phosphorothioate or stereorandom mesyl phosphoramidate intemucleoside linkage.

[0086] As used herein, a “strand” or “strand of linked nucleosides” means contiguous linked nucleosides connected via intemucleoside linkages. A strand of linked nucleosides has a nucleobase sequence. CHEMO 113W0

[0087] As used herein, “subject” means a human or a non-human animal. The subject may be a human. As used herein, “sugar moiety” means an unmodified sugar moiety or a modified sugar moiety.

[0088] As used herein, “sugar surrogate” means a group of atoms other than a furanosyl ring and optionally carrying a nucleobase, and approximately having the size of a furanosyl ring. In certain embodiments, a sugar surrogate is selected from a cyclic sugar surrogate, an acyclic sugar surrogate, and an abasic nucleoside.

[0089] As used herein, “sugar surrogate nucleoside” means a sugar surrogate and optionally a nucleobase, and also includes abasic nucleosides.

[0090] As used herein, “symptom” of a disease means any manifestation, indication, sign, or evidence of a disease. Symptoms include subjective and objective indicia of a disease and may be perceived, experienced, detected, observed, measured, and / or quantified. A symptom may be apparent only upon invasive diagnostic testing, including, but not limited to, post-mortem tests. A symptom may be an absence of a feature, such as failing to reach expected developmental milestones. As used herein, “target nucleic acid” means a nucleic acid that an antisense oligonucleotide is designed to affect. In certain embodiments, the nucleobase sequence of a modified oligonucleotide is complementary to a target nucleic acid. As used herein, “target RNA” means an RNA transcript and includes pre-mRNA and / or mRNA unless otherwise specified.

[0091] As used herein, “target region” refers to a portion of a target nucleic acid that is complementary to the targeting region of an antisense oligonucleotide.

[0092] As used herein, “treating,” or “treatment,” with respect to a disease, means administering a compound or agent to a subject having or at risk for developing such disease. Treating a disease may result in amelioration of at least one symptom of such disease. Treatment may reduce, improve, and / or prevent one or more symptom(s) such that a symptom of the disease is diminished, is no longer apparent, or is never apparent.

[0093] As used herein, “therapeutically effective amount” means an amount of a pharmaceutical agent or composition that provides a therapeutic benefit to a subject. For example, a therapeutically effective amount ameliorates at least one symptom of a disease.

[0094] As used herein, “unmodified nucleobase” means unmodified adenine (A), unmodified thymine (T), unmodified cytosine (C), unmodified uracil (U), or unmodified guanine (G).

[0095] As used herein, an “unmodified nucleoside” means a compound or subunit comprising an unmodified sugar moiety and an unmodified nucleobase.

[0096] As used herein, “unmodified sugar moiety” means a 2'-0H(H) furanosyl sugar moiety, (an “unmodified RNA sugar moiety”), or a 2'-H(H) sugar moiety (an “unmodified DNA sugar moiety”). CHEMO 113W0

[0097] EMBODIMENTS

[0098] Embodiment 1. An oligomeric agent for use in treatment of a disease or condition comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0099] Ab-Wl-G-W2

[0100] I wherein Ab is a bicyclic adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;

[0101] G consists of 5-20 linked 2’-deoxynucleosides;

[0102] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; wherein the oligomeric agent has improved metabolic stability compared to an analogous oligomeric agent having a non-bicyclic nucleoside 5’ of Wl; optionally wherein the modified oligonucleotide consists of the region of formula I.

[0103] Embodiment 2. An oligomeric agent for increasing metabolic stability comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0104] Ab-Wl-G-W2

[0105] I wherein Ab is a bicyclic adenosine; wherein Wl consists of 1-7 linked nucleosides, wherein each nucleoside of Wl is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of Wl is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;

[0106] G consists of 5-20 linked 2’-deoxynucleosides; CHEMO 113W0

[0107] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; provided that the modified oligonucleotide comprises at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate; optionally wherein the modified oligonucleotide consists of the region of formula I.

[0108] Embodiment 3. An oligomeric agent for increasing metabolic stability comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0109] Ab-Wl-G-W2

[0110] I wherein Ab is a bicyclic adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside, and provided that at least one nucleoside of W1 is not a cEt;

[0111] G consists of 5-20 linked 2’-deoxynucleosides;

[0112] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; wherein the modified oligonucleotide comprises at least one phosphodiester intemucleoside linkage; optionally wherein the modified oligonucleotide consists of the region of formula I.

[0113] Embodiment 4. An oligomeric agent for increasing metabolic stability comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0114] Ab-Wl-G-W2

[0115] I wherein Ab is a bicyclic adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified CHEMO 113W0 furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;

[0116] G consists of 11-20 linked 2’-deoxynucleosides;

[0117] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; optionally wherein the modified oligonucleotide consists of the region of formula I.

[0118] Embodiment 5. An oligomeric agent for increasing metabolic stability comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0119] Ab-Wl-G-W2

[0120] I wherein Ab is a bicyclic adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside, and provided that W1 does not comprise a cEt nucleoside;

[0121] G consists of 5-20 linked 2’-deoxynucleosides;

[0122] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; provided that the modified oligonucleotide is not compound 566830; optionally wherein the modified oligonucleotide consists of the region of formula I.

[0123] Embodiment 6. An oligomeric agent for increasing metabolic stability comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0124] Ab-Wl-G-W2

[0125] I CHEMO 113W0 wherein Ab is a bicyclic adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;

[0126] G consists of 5-20 linked 2’-deoxynucleosides;

[0127] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; provided that the modified oligonucleotide comprises at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate; and provided that the modified oligonucleotide comprises either a) or b): a) no more than three 2’-M0E nucleosides; b) no more than three cEt nucleosides; optionally wherein the modified oligonucleotide consists of the region of formula I.

[0128] Embodiment 7. An oligomeric agent for increasing metabolic stability comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0129] Ab-Wl-G-W2

[0130] I wherein Ab is a bicyclic adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;

[0131] G consists of 5-20 linked 2’-deoxynucleosides;

[0132] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside, and provided that W2 comprises at least two non-cEt nucleosides; CHEMO 113W0 provided that the modified oligonucleotide comprises at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate; optionally wherein the modified oligonucleotide consists of the region of formula I.

[0133] Embodiment 8. An oligomeric agent for increasing metabolic stability comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0134] Ab-Wl-G-W2

[0135] I wherein Ab is a bicyclic adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified fiiranosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified fiiranosyl sugar moiety or a sugar surrogate nucleoside;

[0136] G consists of 5-20 linked 2’-deoxynucleosides;

[0137] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified fiiranosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified fiiranosyl sugar moiety or a sugar surrogate nucleoside; provided that the modified oligonucleotide comprises at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate; and further provided that W1 or W2 comprises at least one 2-deoxynucleoside; optionally wherein the modified oligonucleotide consists of the region of formula I.

[0138] Embodiment 9. The oligomeric agent of any preceding embodiment, wherein the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to an Lpa transcript (SEQ ID NO: 1).

[0139] Embodiment 10. The oligomeric agent of any preceding embodiment, wherein the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to an APOE transcript (SEQ ID NO: 2).

[0140] Embodiment 11. The oligomeric agent of any preceding embodiment, wherein the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to a PLN transcript (SEQ ID NO: 3). CHEMO 113W0

[0141] Embodiment 12. The oligomeric agent of any preceding embodiment, wherein Ab is a cEt adenosine.

[0142] Embodiment 13. The oligomeric agent of any preceding embodiment, wherein Ab is an LNA adenosine.

[0143] Embodiment 14. The oligomeric agent of any preceding embodiment, wherein each nucleoside of W1 is independently selected from a bicyclic nucleoside other than a cEt nucleoside, a 2’- substituted nucleoside, a sugar surrogate nucleoside, and a 2’-deoxynucleoside.

[0144] Embodiment 15. The oligomeric agent of any preceding embodiment, comprising at least one phosphorothioate intemucleoside linkage.

[0145] Embodiment 16. The oligomeric agent of any preceding embodiment, comprising at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate.

[0146] Embodiment 17. The oligomeric agent of any preceding embodiment, wherein the 3 ’-most nucleoside of W1 does not comprise a 2’-0Me nucleoside.

[0147] Embodiment 18. The oligomeric agent of any preceding embodiment, wherein the modified oligonucleotide comprises no more than three cEt nucleosides.

[0148] Embodiment 19. The oligomeric agent of any preceding embodiment, wherein the modified oligonucleotide comprises no more than three 2’-M0E nucleosides.

[0149] Embodiment 20. The oligomeric agent of any preceding embodiment, wherein the modified oligonucleotide comprises at least four 2’-MOE nucleosides.

[0150] Embodiment 21. The oligomeric agent of any preceding embodiment, wherein W 1 comprises no more than three 2 ’-MOE nucleosides.

[0151] Embodiment 22. The oligomeric agent of any preceding embodiment, wherein W1 does not comprise three contiguous cEt nucleosides.

[0152] Embodiment 23. The oligomeric agent of any preceding embodiment, wherein W1 does not comprise three contiguous 2 ’-MOE nucleosides.

[0153] Embodiment 24. The oligomeric agent of any preceding embodiment, wherein each intemucleoside linkage at the 3 ’-position of each nucleoside of G is independently selected from a mesyl phosphoramidate intemucleoside linkage and a phosphorothioate intemucleoside linkage.

[0154] Embodiment 25. The oligomeric agent of any one of the preceding embodiments, wherein the modified oligonucleotide comprises at least one mesyl phosphoramidate intemucleoside linkage. CHEMO 113W0

[0155] Embodiment 26. The oligomeric agent of any one of the preceding embodiments, wherein each nucleoside of W1 is independently selected from a 2 ’-substituted nucleoside, a bicyclic nucleoside, and a 2 ’-deoxynucleoside.

[0156] Embodiment 27. The oligomeric agent of any one of the preceding embodiments, wherein each nucleoside of W1 is independently selected from a 2 ’-substituted nucleoside, a bicyclic nucleoside other than a cEt nucleoside, and a 2 ’-deoxynucleoside.

[0157] Embodiment 28. The oligomeric agent of any one of the preceding embodiments, wherein each nucleoside of W1 is independently selected from a 2 ’-substituted nucleoside and a 2’- deoxynucleoside.

[0158] Embodiment 29. The oligomeric agent of any one of the preceding embodiments, wherein each nucleoside of W1 is independently a 2 ’-substituted nucleoside or a bicyclic nucleoside other than a cEt nucleoside.

[0159] Embodiment 30. The oligomeric agent of any one of the preceding embodiments, wherein the 3’- most nucleoside ofWl is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage.

[0160] Embodiment 31. The oligomeric agent of any one of the preceding embodiments, wherein G consists of at least 11 linked nucleosides, optionally 11, 12, 13, 14, or 15 linked nucleosides.

[0161] Embodiment 32. The oligomeric agent of any one of the preceding embodiments, wherein G consists of 10 linked nucleosides.

[0162] Embodiment 33. The oligomeric agent of any one of the preceding embodiments, wherein the modified oligonucleotide comprises one, two, or three cEt nucleosides.

[0163] Embodiment 34. The oligomeric agent of any one of the preceding embodiments, wherein at least one of W1 and W2 comprises a 2’ -deoxynucleoside.

[0164] Embodiment 35. The oligomeric agent of any preceding embodiment, wherein each 2’- deoxynucleoside of W1 and W2 is linked at its respective 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage.

[0165] Embodiment 36. The oligomeric agent of any preceding embodiment, wherein each 2’- deoxynucleoside of W1 is linked at its 3’-position by a mesyl phosphoramidate intemucleoside linkage.

[0166] Embodiment 37. The oligomeric agent of any preceding embodiment, wherein each 2’- deoxynucleoside of W2 is linked at its 3’-position by a mesyl phosphoramidate intemucleoside linkage. CHEMO 113W0

[0167] Embodiment 38. The oligomeric agent of any preceding embodiment, wherein W1 comprises 1-4 modified nucleosides comprising a modified furanosyl sugar moiety.

[0168] Embodiment 39. The oligomeric agent of any preceding embodiment, wherein W1 comprises 2 or 3 modified nucleosides comprising a modified furanosyl sugar moiety, optionally 3.

[0169] Embodiment 40. The oligomeric agent of any preceding embodiment, wherein W1 comprises 3 contiguous modified nucleosides comprising a modified furanosyl sugar moiety.

[0170] Embodiment 41. The oligomeric agent of any preceding embodiment, wherein each modified nucleoside of W1 is linked at its 3’-position by a phosphodiester or a phosphorothioate intemucleoside linkage, optionally a phosphorothioate intemucleoside linkage.

[0171] Embodiment 42. The oligomeric agent of any preceding embodiment, wherein the 3 ’-most nucleoside of W1 is linked at its 3 ’-position by a phosphorothioate intemucleoside linkage.

[0172] Embodiment 43. The oligomeric agent of any preceding embodiment, wherein W1 consists of 2, 3, or 4 linked nucleosides.

[0173] Embodiment 44. The oligomeric agent of any preceding embodiment, wherein W1 does not comprise a 2 ’-deoxynucleoside.

[0174] Embodiment 45. The oligomeric agent of any one of embodiments 1-43, wherein W1 comprises at least one 2 ’-deoxynucleoside.

[0175] Embodiment 46. The oligomeric agent of any preceding embodiment, wherein each 2 ’-substituted nucleoside of W1 is a 2 ’-MOE nucleoside.

[0176] Embodiment 47. The oligomeric agent of any preceding embodiment, wherein W 1 comprises at least four 2’-MOE nucleosides.

[0177] Embodiment 48. The oligomeric agent of any preceding embodiment, wherein each nucleoside of W1 is a 2 ’-MOE nucleoside.

[0178] Embodiment 49. The oligomeric agent of any preceding embodiment, wherein W1 consists of four contiguous 2’-MOE nucleosides.

[0179] Embodiment 50. The oligomeric agent of any preceding embodiment, wherein each 2’- deoxynucleoside of W1 is a 2’-P-D-deoxyribosyl nucleoside.

[0180] Embodiment 51. The oligomeric agent of any preceding embodiment, wherein each bicyclic nucleoside of W1 is an LNA nucleoside.

[0181] Embodiment 52. The oligomeric agent of any preceding embodiment, wherein W2 comprises 1-5 modified nucleosides comprising a modified furanosyl sugar moiety. CHEMO 113W0

[0182] Embodiment 53. The oligomeric agent of any preceding embodiment, wherein W2 comprises 3 modified nucleosides comprising a modified furanosyl sugar moiety.

[0183] Embodiment 54. The oligomeric agent of any preceding embodiment, wherein W2 comprises 5 modified nucleosides comprising a modified furanosyl sugar moiety.

[0184] Embodiment 55. The oligomeric agent of any preceding embodiment, wherein each non-terminal modified nucleoside of W2 is linked at its 3’-position by a phosphodiester or a phosphorothioate intemucleoside linkage, optionally a phosphorothioate intemucleoside linkage.

[0185] Embodiment 56. The oligomeric agent of any preceding embodiment, wherein W2 consists of 3, 4, 5, or 6 linked nucleosides.

[0186] Embodiment 57. The oligomeric agent of any preceding embodiment, wherein W2 consists of 3, 4, or 5 linked nucleosides.

[0187] Embodiment 58. The oligomeric agent of any preceding embodiment, wherein W2 does not comprise a 2 ’-deoxynucleoside.

[0188] Embodiment 59. The oligomeric agent of any of embodiments 1-57, wherein W2 comprises at least one 2 ’-deoxynucleoside.

[0189] Embodiment 60. The oligomeric agent of any preceding embodiment, wherein each modified nucleoside of W2 is independently selected from a 2 ’-substituted nucleoside and a bicyclic nucleoside.

[0190] Embodiment 61. The oligomeric agent of any preceding embodiment, wherein each modified nucleoside of W2 is independently selected from a 2 ’-substituted nucleoside and a bicyclic nucleoside other than a cEt nucleoside.

[0191] Embodiment 62. The oligomeric agent of any preceding embodiment, wherein each modified nucleoside ofW2 is independently a bicyclic nucleoside.

[0192] Embodiment 63. The oligomeric agent of any preceding embodiment, wherein each bicyclic nucleoside of W2 is selected from cEt and LNA.

[0193] Embodiment 64. The oligomeric agent of any preceding embodiment, wherein each modified nucleoside ofW2 is independently a 2’ -substituted nucleoside.

[0194] Embodiment 65. The oligomeric agent of any preceding embodiment, wherein each 2 ’-substituted nucleoside of W2 is a 2 ’-MOE nucleoside.

[0195] Embodiment 66. The oligomeric agent of any preceding embodiment, wherein each 2’- deoxynucleoside of W2 is a 2’-P-D-deoxyribosyl nucleoside. CHEMO 113W0

[0196] Embodiment 67. The oligomeric agent of any preceding embodiment, wherein each nucleoside of G is a 2’-P-D-deoxyribosyl nucleoside.

[0197] Embodiment 68. The oligomeric agent of any preceding embodiment, wherein each nucleoside of G is linked at its 3 ’-position by a phosphorothioate or a mesyl phosphoramidate intemucleoside linkage.

[0198] Embodiment 69. The oligomeric agent of any preceding embodiment, wherein at least one, optionally one, nucleoside of G is linked at its 3 ’-position by a mesyl phosphoramidate intemucleoside linkage.

[0199] Embodiment 70. The oligomeric agent of any preceding embodiment, wherein two, three, four, five, or six nucleosides of G are linked at their respective 3 ’-positions by a mesyl phosphoramidate intemucleoside linkage.

[0200] Embodiment 71. The oligomeric agent of any preceding embodiment, wherein five, six, seven, eight, nine, or ten nucleosides of G are linked at their respective 3 ’-positions by a phosphorothioate intemucleoside linkage.

[0201] Embodiment 72. The oligomeric agent of any preceding embodiment, wherein the 3 ’-most nucleoside of G is linked at its 3 ’-position by a mesyl phosphoramidate or phosphorothioate intemucleoside linkage.

[0202] Embodiment 73. The oligomeric agent of any preceding embodiment, wherein G consists of 7-15 linked nucleosides, optionally 10-13 linked nucleosides, optionally exactly 10 linked nucleosides.

[0203] Embodiment 74. The oligomeric agent of any preceding embodiment, wherein G is a deoxy region.

[0204] Embodiment 75. The oligomeric agent of any preceding embodiment, wherein W2 has the formula -(E)n4-(D4)n5-(F)n6 wherein each F is independently selected from a modified nucleoside comprising a modified fiiranosyl sugar moiety, a sugar surrogate nucleoside, and a 2’ -deoxy nucleoside, provided that at least one F is a modified nucleoside comprising a modified fiiranosyl sugar moiety or a sugar surrogate nucleoside; and each non-terminal 2’-deoxy nucleoside of F is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; each E is independently selected from a modified nucleoside comprising a modified fiiranosyl sugar moiety and a sugar surrogate nucleoside; each D4 is independently a 2’-deoxy nucleoside provided that each D4 is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; n6 is 1 to 8 and each of n4 and n5 is independently 0 to 8.

[0205] Embodiment 76. The oligomeric agent of embodiment 75, wherein n4 is 1 or 2. CHEMO 113W0

[0206] Embodiment 77. The oligomeric agent of embodiment 75 or 76, wherein n5 is 1.

[0207] Embodiment 78. The oligomeric agent of embodiment 75 or 76, wherein n5 is 0.

[0208] Embodiment 79. The oligomeric agent of any one of embodiments 75-78, wherein n6 is 1 to 5.

[0209] Embodiment 80. The oligomeric agent of any one of embodiments 75-79, wherein n6 is 1, 2, or 3.

[0210] Embodiment 81. The oligomeric agent of any one of embodiments 75-80, wherein each E is selected from a bicyclic nucleoside and a 2 ’-substituted nucleoside.

[0211] Embodiment 82. The oligomeric agent of any one of embodiments 75-81, wherein each E is a bicyclic nucleoside.

[0212] Embodiment 83. The oligomeric agent of any one of embodiments 75-82, wherein each bicyclic nucleoside of E is selected from cEt and LNA, optionally cEt.

[0213] Embodiment 84. The oligomeric agent of any one of embodiments 75-81, wherein each E is a 2’- substituted nucleoside.

[0214] Embodiment 85. The oligomeric agent of any one of embodiments 75-84, wherein each 2’- substituted nucleoside of E is a 2’-MOE nucleoside.

[0215] Embodiment 86. The oligomeric agent of any one of embodiments 75-85, wherein each D4 is a 2’- P-D-deoxyribosyl nucleoside.

[0216] Embodiment 87. The oligomeric agent of any one of embodiments 75-86, wherein each F is selected from a bicyclic nucleoside and a 2 ’-substituted nucleoside.

[0217] Embodiment 88. The oligomeric agent of any one of embodiments 75-87, wherein each F is a 2’- substituted nucleoside.

[0218] Embodiment 89. The oligomeric agent of any one of embodiments 75-88, wherein 2 ’-substituted nucleoside of F is a 2’-MOE nucleoside.

[0219] Embodiment 90. The oligomeric agent of any one of embodiments 75-87 or 89, wherein each F is a bicyclic nucleoside.

[0220] Embodiment 91. The oligomeric agent of any one of embodiments 75-90, wherein each bicyclic nucleoside of F is selected from cEt and LNA, optionally cEt.

[0221] Embodiment 92. The oligomeric agent of any one of embodiments 75-91, wherein each of n4 and n5 is 0.

[0222] Embodiment 93. The oligomeric agent of any one of embodiments 75-92, wherein n6 is 5, and each of n4 and n5 is 0. CHEMO 113W0

[0223] Embodiment 94. The oligomeric agent of any preceding embodiment, wherein W 1 has the formula (A)nl-(Dl)n2-(B)n3- wherein each A is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’ -deoxy nucleoside provided that at least one A is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; and each 2’-deoxy nucleoside of A is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; each B is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety and a sugar surrogate nucleoside; each DI is independently a 2’-deoxy nucleoside provided that each DI is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; nl is 1 to 8 and each of n2 and n3 is independently 0 to 8.

[0224] Embodiment 95. The oligomeric agent of embodiment 94, wherein n3 is 0 or 1.

[0225] Embodiment 96. The oligomeric agent of embodiment 94 or 95, wherein n2 is 0 or 1.

[0226] Embodiment 97. The oligomeric agent of any one of embodiments 94-96, wherein nl is 1 to 5.

[0227] Embodiment 98. The oligomeric agent of any one of embodiments 94-97, wherein nl is 2 or 3.

[0228] Embodiment 99. The oligomeric agent of any one of embodiments 94-98, wherein each B is selected from a bicyclic nucleoside and a 2 ’-substituted nucleoside.

[0229] Embodiment 100. The oligomeric agent of any one of embodiments 94-99, wherein each B is a 2’- substituted nucleoside.

[0230] Embodiment 101. The oligomeric agent of any one of embodiments 94-100, wherein each 2’- substituted nucleoside of B is a 2’-MOE nucleoside.

[0231] Embodiment 102. The oligomeric agent of any one of embodiments 94-101, wherein each B is a bicyclic nucleoside.

[0232] Embodiment 103. The oligomeric agent of any one of embodiments 94-102, wherein each bicyclic nucleoside of B is selected from cEt and LNA, optionally cEt.

[0233] Embodiment 104. The oligomeric agent of any one of embodiments 94-103, wherein each D4 is a 2’-P-D-deoxyribosyl nucleoside.

[0234] Embodiment 105. The oligomeric agent of any one of embodiments 94-104, wherein each A is selected from a bicyclic nucleoside and a 2 ’-substituted nucleoside.

[0235] Embodiment 106. The oligomeric agent of any one of embodiments 94-105, wherein each A is a 2’- substituted nucleoside. CHEMO 113W0

[0236] Embodiment 107. The oligomeric agent of any one of embodiments 94-106, wherein each 2’- substituted nucleoside of A is a 2 ’-MOE nucleoside.

[0237] Embodiment 108. The oligomeric agent of any one of embodiments 94-105 or 107, wherein each A is a bicyclic nucleoside.

[0238] Embodiment 109. The oligomeric agent of any one of embodiments 94-108, wherein each bicyclic nucleoside of A is selected from cEt and LNA, optionally cEt.

[0239] Embodiment 110. The oligomeric agent of any one of embodiments 94-109, wherein each of n2 and n3 is 0.

[0240] Embodiment 111. The oligomeric agent of any one of embodiments 94-110, wherein nl is 4, and each of n2 and n3 is 0.

[0241] Embodiment 112. The oligomeric agent of any preceding embodiment, wherein the modified oligonucleotide consists of 17-20 linked nucleosides.

[0242] Embodiment 113. The oligomeric agent of any preceding embodiment, wherein the modified oligonucleotide consists of 17, 18, 19, or 20 linked nucleosides.

[0243] Embodiment 114. The oligomeric agent of any preceding embodiment, wherein the modified oligonucleotide consists of 20 linked nucleosides.

[0244] Embodiment 115. The oligomeric agent of any one of embodiments 1-114, wherein G consists of 11-15 linked nucleosides, optionally 11, optionally 12, optionally 13, optionally 14, or optionally 15 linked nucleosides, optionally wherein neither W1 nor W2 comprises a 2 ’-deoxynucleoside.

[0245] Embodiment 116. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0246] Ak-Wl-G-W2

[0247] I wherein Akis a cEt adenosine, W1 consists of 4 linked nucleosides, wherein each nucleoside of W1 is selected from a 2 ’-substituted nucleoside and a bicyclic nucleoside;

[0248] G consists of 10 linked 2'-P-D-deoxyribosyl nucleosides;

[0249] W2 consists of 5 linked nucleosides, wherein each nucleoside of W2 is independently selected from a 2 ’-substituted nucleoside and a bicyclic nucleoside; optionally wherein the modified oligonucleotide consists of the region of formula I. CHEMO 113W0

[0250] Embodiment 117. The oligomeric agent of embodiment 116, wherein W1 comprises exactly one bicyclic nucleoside.

[0251] Embodiment 118. The oligomeric agent of embodiment 116, wherein each nucleoside of W1 is 2’- substituted nucleoside.

[0252] Embodiment 119. The oligomeric agent of any one of embodiments 116-118, wherein each 2’- substituted nucleoside of W1 is selected from a 2 ’-MOE nucleoside and a 2’-OMe nucleoside.

[0253] Embodiment 120. The oligomeric agent of embodiment 119, wherein each 2 ’-substituted nucleoside of W1 is a 2 ’-MOE nucleoside.

[0254] Embodiment 121. The oligomeric agent of any one of embodiments 116-120, wherein each bicyclic nucleoside of W1 is selected from a cEt nucleoside and an LNA nucleoside.

[0255] Embodiment 122. The oligomeric agent of any one of embodiments 116-121, wherein each bicyclic nucleoside of W1 is a cEt nucleoside.

[0256] Embodiment 123. The oligomeric agent of any one of embodiments 116-122, wherein W2 comprises no bicyclic nucleosides.

[0257] Embodiment 124. The oligomeric agent of any one of embodiments 116-122, wherein W2 comprises exactly one bicyclic nucleoside.

[0258] Embodiment 125. The oligomeric agent of any one of embodiments 116-122, wherein W2 comprises exactly two bicyclic nucleosides.

[0259] Embodiment 126. The oligomeric agent of any one of embodiments 116-125, wherein each 2’- substituted nucleoside of W2 is selected from a 2 ’-MOE nucleoside and a 2’-OMe nucleoside.

[0260] Embodiment 127. The oligomeric agent of embodiment 126, wherein each 2 ’-substituted nucleoside of W2 is a 2 ’-MOE nucleoside.

[0261] Embodiment 128. The oligomeric agent of any one of embodiments 116-127, wherein each intemucleoside linkage is selected from a phosphodiester, a phosphorothioate, and a mesyl phosphoramidate .

[0262] Embodiment 129. The oligomeric agent of any preceding embodiment, wherein the modified oligonucleotide comprises nucleobases independently selected from thymine, uracil, guanine, cytosine, 5 -methylcytosine, adenine, and hypoxanthine.

[0263] Embodiment 130. The oligomeric agent of any preceding embodiment, comprising a conjugate group.

[0264] Embodiment 131. The oligomeric agent of embodiment 130, wherein the conjugate group comprises a carbohydrate or carbohydrate cluster. CHEMO 113W0

[0265] Embodiment 132. The oligomeric agent of embodiment 130 or 131, wherein the conjugate group comprises at least one GalNAc.

[0266] Embodiment 133. The oligomeric agent of embodiment 130, wherein the conjugate group comprises a C10-C20 alkyl chain.

[0267] Embodiment 134. The oligomeric agent of embodiment 133, wherein the conjugate group comprises Cl 6 alkyl.

[0268] Embodiment 135. The oligomeric agent of embodiment 130, wherein the conjugate group comprises a transferrin receptor ligand.

[0269] Embodiment 136. The oligomeric agent of embodiment 135, wherein the transferrin receptor ligand comprises a peptide.

[0270] Embodiment 137. The oligomeric agent of embodiment 135 or 136, wherein the transferrin receptor ligand comprises a bicyclic peptide.

[0271] Embodiment 138. The oligomeric agent of embodiment 135, wherein the transferrin receptor ligand comprises a Fab, Fab’, F(ab’)2, scFv, VHH, VNAR, or nanobody.

[0272] Embodiment 139. The oligomeric agent of any one of embodiments 130-138, wherein the oligomeric agent comprises the modified oligonucleotide, the conjugate group, and a cleavable linker comprising nucleosides linked by phosphodiester linkages linking the modified oligonucleotide and the conjugate group.

[0273] Embodiment 140. The oligomeric agent of any one of embodiments 1-139, wherein the oligomeric agent consists of the modified oligonucleotide.

[0274] Embodiment 141. The oligomeric agent of any preceding embodiment, wherein the nucleobase sequence of the modified oligonucleotide is complementary to a target nucleic acid.

[0275] Embodiment 142. The oligomeric agent of embodiment 141, wherein the nucleobase sequence of the modified oligonucleotide is at least 80%, at least 90%, or 100% complementary to the target nucleic acid.

[0276] Embodiment 143. The oligomeric agent of embodiment 141 or 142, wherein the target nucleic acid is selected from: an mRNA, a pre-mRNA, a microRNA, and a non-coding RNA.

[0277] Embodiment 144. The oligomeric agent of any preceding embodiment, wherein the modified oligonucleotide is a first modified oligonucleotide, and further comprising a second modified oligonucleotide that is complementary to the first modified oligonucleotide.

[0278] Embodiment 145. A pharmaceutical composition comprising the oligomeric agent of any preceding embodiment and a pharmaceutically acceptable carrier. CHEMO 113W0

[0279] Embodiment 146. A method comprising contacting a cell with the oligomeric agent or pharmaceutical composition of any preceding embodiment.

[0280] Embodiment 147. A method of modulating the amount or activity of a target nucleic acid in a cell, comprising contacting the cell with the oligomeric agent of any of embodiments 1-144 or pharmaceutical composition of embodiment 145.

[0281] Embodiment 148. The method of embodiment 146-147, whereby the amount or activity of a target nucleic acid is reduced.

[0282] Embodiment 149. The method of any one of embodiments 146-148, comprising parenteral administration to a subject.

[0283] Embodiment 150. The method of any one of embodiments 146-149, comprising injection into CSF.

[0284] Embodiment 151. The method of any one of embodiments 146-149, comprising subcutaneous injection.

[0285] Embodiment 152. The oligomeric agent of any of embodiments 1-144 or the pharmaceutical composition of embodiment 145, for use in treatment of a disease or condition.

[0286] Embodiment 153. The oligomeric agent of any of embodiments 1-144 or the pharmaceutical composition of embodiment 145, for use in the manufacture of a medicament.

[0287] Embodiment 154. The oligomeric agent of any of embodiments 1-144 or the pharmaceutical composition of embodiment 145, for use in therapy.

[0288] Embodiment 155. An oligomeric agent for use in treatment of a disease or condition comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0289] Ab-Wl-G-W2

[0290] I wherein Ab is a bicyclic adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;

[0291] G consists of 5-20 linked 2’-deoxynucleosides;

[0292] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate CHEMO 113W0 nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; wherein the oligomeric agent has improved metabolic stability compared to an analogous oligomeric agent having a non-bicyclic nucleoside 5’ of Wl; optionally wherein the modified oligonucleotide consists of the region of formula I.

[0293] Embodiment 156. The oligomeric agent of embodiment 155, wherein at least one of Wl and / or W2 comprises a 2 ’-deoxynucleoside and at least one intemucleoside linkage is not a phosphorothioate or phosphodiester.

[0294] Embodiment 157. The oligomeric agent of embodiment 156, wherein the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to an Lpa transcript (SEQ ID NO: 1).

[0295] Embodiment 158. The oligomeric agent of embodiment 155, wherein at least one of Wl and / or W2 comprises a 2’-MOE nucleoside and at least one intemucleoside linkage is a phosphodiester.

[0296] Embodiment 159. The oligomeric agent of embodiment 155, wherein at least one of Wl and / or W2 comprises a 2 ’-MOE nucleoside and the modified oligonucleotide comprises an intemucleoside linkage other than a phosphorothioate, phosphodiester, or a methoxypropyl phosphonate linkage.

[0297] Embodiment 160. The oligomeric agent of embodiment 155, wherein at least one of Wl and / or W2 comprises a 2’-MOE nucleoside and the modified oligonucleotide comprises a mesyl phosphoramidate intemucleoside linkage.

[0298] Embodiment 161. The oligomeric agent of any of embodiment 158-160, wherein the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to an APOE transcript (SEQ ID NO: 2).

[0299] Embodiment 162. The oligomeric agent of embodiment 155, wherein the 5’-most intemucleoside linkage is a mesyl phosphoramidate intemucleoside linkage, and wherein the modified oligonucleotide comprises at least one modified nucleoside comprising a modified furanosyl sugar moiety other than a cEt sugar moiety or at least one sugar surrogate nucleoside.

[0300] Embodiment 163. The oligomeric agent of embodiment 155, wherein the 5’-most intemucleoside linkage is a mesyl phosphoramidate intemucleoside linkage, and wherein the modified oligonucleotide comprises no more than 3 cEt nucleosides.

[0301] Embodiment 164. The oligomeric agent of embodiment 155, wherein the 5’-most intemucleoside linkage is a mesyl phosphoramidate intemucleoside linkage;

[0302] W 1 is not two consecutive cEt nucleosides, and CHEMO 113W0 wherein the modified oligonucleotide comprises no more than 9 mesyl phosphoramidate intemucleoside linkage.

[0303] Embodiment 165. The oligomeric agent of embodiment 155, wherein Ab comprises a cEt sugar moiety, and G is from 11-20.

[0304] Embodiment 166. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0305] Ab-Wl-G-W2

[0306] I wherein Ab is a bicyclic adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified fiiranosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified fiiranosyl sugar moiety or a sugar surrogate nucleoside;

[0307] G consists of 5-20 linked 2’-deoxynucleosides;

[0308] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified fiiranosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified fiiranosyl sugar moiety or a sugar surrogate nucleoside; provided that the modified oligonucleotide comprises at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate; and provided that the modified oligonucleotide does not comprise a methylphosphonate or methoxypropyl phosphonate intemucleoside linkage; optionally wherein the modified oligonucleotide consists of the region of formula L

[0309] Embodiment 167. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0310] Ab-Wl-G-W2

[0311] I wherein Ab is a cEt adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified fiiranosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the CHEMO 113W0

[0312] 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside, and provided that at least one nucleoside of W1 is not a cEt;

[0313] G consists of 5-20 linked 2’-deoxynucleosides;

[0314] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; wherein the modified oligonucleotide comprises at least one phosphodiester intemucleoside linkage; optionally wherein the modified oligonucleotide consists of the region of formula I.

[0315] Embodiment 168. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0316] Ab-Wl-G-W2

[0317] I wherein Ab is a cEt adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;

[0318] G consists of 11-20 linked 2’-deoxynucleosides;

[0319] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; optionally wherein the modified oligonucleotide consists of the region of formula I.

[0320] Embodiment 169. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0321] Ab-Wl-G-W2

[0322] I CHEMO 113W0 wherein Ab is a cEt adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside, and provided that W1 does not comprise a cEt nucleoside;

[0323] G consists of 5-20 linked 2’-deoxynucleosides;

[0324] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; provided that the modified oligonucleotide comprises at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate; optionally wherein the modified oligonucleotide consists of the region of formula I.

[0325] Embodiment 170. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0326] Ab-Wl-G-W2

[0327] I wherein Ab is a bicyclic adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;

[0328] G consists of 5-20 linked 2’-deoxynucleosides;

[0329] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; provided that the modified oligonucleotide comprises at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate; and provided that the modified oligonucleotide comprises at least one but no more than 3 2’- MOE nucleosides and W1 is not 2 consecutive cEt nucleosides; optionally wherein the modified oligonucleotide consists of the region of formula I. CHEMO 113W0

[0330] Embodiment 171. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0331] Ab-Wl-G-W2

[0332] I wherein Ab is a cEt adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;

[0333] G consists of 5-20 linked 2’-deoxynucleosides;

[0334] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside, and provided that W2 comprises at least two non-cEt nucleosides; provided that the modified oligonucleotide comprises at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate; and the modified oligonucleotide does not comprise a methoxy propyl phosphonate intemucleoside linkage; optionally wherein the modified oligonucleotide consists of the region of formula I.

[0335] Embodiment 172. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0336] Ab-Wl-G-W2

[0337] I wherein Ab is a bicyclic adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;

[0338] G consists of 5-20 linked 2’-deoxynucleosides; CHEMO 113W0

[0339] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; provided that the modified oligonucleotide comprises at least one phosphodiester intemucleoside linkage; and further provided that W1 or W2 comprises at least one 2-deoxynucleoside; optionally wherein the modified oligonucleotide consists of the region of formula I.

[0340] Embodiment 173. The oligomeric agent of any of embodiments 166-18, wherein the oligomeric agent has improved metabolic stability compared to an analogous oligomeric agent having a non- bicyclic nucleoside 5’ ofWl.

[0341] Embodiment 174. The oligomeric agent of any of embodiments 155-173, wherein the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to an Lpa transcript (SEQ ID NO: 1).

[0342] Embodiment 175. The oligomeric agent of any of embodiments 155-174, wherein the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to an APOE transcript (SEQ ID NO: 2).

[0343] Embodiment 176. The oligomeric agent of any of embodiments 155-175, wherein the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to a PLN transcript (SEQ ID NO: 3).

[0344] Embodiment 177. The oligomeric agent of any of embodiments 155-164, 166, or 170 wherein Ab is a cEt adenosine.

[0345] Embodiment 178. The oligomeric agent of any of embodiments 155-164, 166, or 170 wherein Ab is an LNA adenosine.

[0346] Embodiment 179. The oligomeric agent of any of embodiments 155-168 or 170-178, wherein each nucleoside of W1 is independently selected from a bicyclic nucleoside other than a cEt nucleoside, a non-bicyclic 2 ’-substituted nucleoside, a sugar surrogate nucleoside, and a 2’- deoxynucleoside.

[0347] Embodiment 180. The oligomeric agent of any of embodiments 155-179, comprising at least one phosphorothioate intemucleoside linkage.

[0348] Embodiment 181. The oligomeric agent of any of embodiments 155-166, 168-171, or 173-180, comprising at least one phosphodiester intemucleoside linkage. G CHEMO 113W0

[0349] Embodiment 182. The oligomeric agent of any of any of embodiments 155-181 , comprising at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate.

[0350] Embodiment 183. The oligomeric agent of any of embodiments 155-182, wherein the modified oligonucleotide does not comprise a methyl propyl phosphonate or a methylphosphonate intemucleoside linkage.

[0351] Embodiment 184. The oligomeric agent of any of embodiments 155-183, wherein the modified oligonucleotide comprises at least one mesyl phosphoramidate intemucleoside linkage.

[0352] Embodiment 185. The oligomeric agent of any of embodiments 155-184, wherein each intemucleoside linkage is independently selected from a mesyl phosphoramidate, a phosphorothioate, and a phosphodiester intemucleoside linkage.

[0353] Embodiment 186. The oligomeric agent of any of embodiments 155-185, wherein the 3’-most nucleoside of W1 does not comprise a 2’-OMe nucleoside.

[0354] Embodiment 187. The oligomeric agent of any embodiments 155-186, wherein the modified oligonucleotide comprises no more than three cEt nucleosides.

[0355] Embodiment 188. The oligomeric agent of any of embodiments 155-187, wherein the modified oligonucleotide comprises no more than three 2’-MOE nucleosides.

[0356] Embodiment 189. The oligomeric agent of any of embodiments 155-169 or 171-188, wherein the modified oligonucleotide comprises at least four 2’-MOE nucleosides.

[0357] Embodiment 190. The oligomeric agent of any of embodiments 155-189, , wherein W1 comprises no more than three 2 ’-MOE nucleosides.

[0358] Embodiment 191. The oligomeric agent of any of embodiments 155-190, wherein W1 does not comprise three contiguous cEt nucleosides.

[0359] Embodiment 192. The oligomeric agent of any of embodiments 155-191, wherein W1 does not comprise two contiguous cEt nucleosides.

[0360] Embodiment 193. The oligomeric agent of any of embodiments 155-192, wherein W1 does not comprise three contiguous 2 ’-MOE nucleosides.

[0361] Embodiment 194. The oligomeric agent of any of embodiments 155-193, wherein each intemucleoside linkage at the 3 ’-position of each nucleoside of G is independently selected from a mesyl phosphoramidate intemucleoside linkage and a phosphorothioate intemucleoside linkage.

[0362] Embodiment 195. The oligomeric agent of any of embodiments 155-194, wherein each nucleoside of W1 is independently selected from a non-bicyclic 2 ’-substituted nucleoside, a bicyclic nucleoside, and a 2 ’-deoxynucleoside. CHEMO 113W0

[0363] Embodiment 196. The oligomeric agent of any of embodiments 155-194, wherein each nucleoside of W1 is independently selected from a non-bicyclic 2 ’-substituted nucleoside, a bicyclic nucleoside other than a cEt nucleoside, and a 2 ’-deoxynucleoside.

[0364] Embodiment 197. The oligomeric agent of any of embodiments 155-194, wherein each nucleoside of W1 is independently selected from a non-bicyclic 2 ’-substituted nucleoside and a 2’- deoxynucleoside.

[0365] Embodiment 198. The oligomeric agent of any of embodiments 155-194, wherein each nucleoside of W1 is independently a non-bicyclic 2 ’-substituted nucleoside or a bicyclic nucleoside other than a cEt nucleoside.

[0366] Embodiment 199. The oligomeric agent of any of embodiments 155-198, wherein the 3’-most nucleoside of W1 is linked at its 3’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage.

[0367] Embodiment 200. The oligomeric agent of any of embodiments 155-199, wherein G consists of at least 11 linked nucleosides, optionally 11, 12, 13, 14, or 15 linked nucleosides.

[0368] Embodiment 201. The oligomeric agent of embodiments 155-164, 166-167, or 169-199, wherein G consists of 10 linked nucleosides.

[0369] Embodiment 202. The oligomeric agent of any of embodiments 155-201, wherein the modified oligonucleotide comprises exactly one, exactly two, or exactly three cEt nucleosides.

[0370] Embodiment 203. The oligomeric agent of any of embodiments 155-202, wherein at least one of W1 and W2 comprises a 2 ’-deoxynucleoside.

[0371] Embodiment 204. The oligomeric agent of any of embodiments 155-203, wherein each 2’- deoxynucleoside of W1 and W2 is linked at its respective 3’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage.

[0372] Embodiment 205. The oligomeric agent of any of embodiments 155-204, wherein each 2’- deoxynucleoside of W1 is linked at its 3’-position by a mesyl phosphoramidate intemucleoside linkage.

[0373] Embodiment 206. The oligomeric agent of any of embodiments 155-205, wherein each 2’- deoxynucleoside of W2 is linked at its 3’-position by a mesyl phosphoramidate intemucleoside linkage.

[0374] Embodiment 207. The oligomeric agent of any of embodiments 155-206, wherein W1 comprises 1- 4 modified nucleosides comprising a modified furanosyl sugar moiety.

[0375] Embodiment 208. The oligomeric agent of any of embodiments 155-206, wherein W1 comprises 2 or 3 modified nucleosides comprising a modified furanosyl sugar moiety, optionally 3. CHEMO 113W0

[0376] Embodiment 209. The oligomeric agent of any of embodiments 155-206, wherein W1 comprises 3 contiguous modified nucleosides comprising a modified furanosyl sugar moiety.

[0377] Embodiment 210. The oligomeric agent of any of embodiments 155-209, wherein each modified nucleoside of W1 is linked at its 3’-position by a phosphodiester or a phosphorothioate intemucleoside linkage, optionally a phosphorothioate intemucleoside linkage.

[0378] Embodiment 211. The oligomeric agent of any of embodiments 155-209, wherein the 3 ’-most nucleoside of W1 is linked at its 3 ’-position by a phosphorothioate intemucleoside linkage.

[0379] Embodiment 212. The oligomeric agent of any of embodiments 155-211, wherein W1 consists of 2, 3, or 4 linked nucleosides.

[0380] Embodiment 213. The oligomeric agent of any of embodiments 155-211, wherein W1 does not comprise a 2 ’-deoxynucleoside.

[0381] Embodiment 214. The oligomeric agent of any of embodiments 155-212, wherein W1 comprises at least one 2 ’-deoxynucleoside.

[0382] Embodiment 215. The oligomeric agent of any of embodiments 155-214, wherein each 2’- substituted nucleoside of W1 is a 2 ’-MOE nucleoside.

[0383] Embodiment 216. The oligomeric agent of any of embodiments 155-215, wherein W1 comprises at least four 2’-M0E nucleosides.

[0384] Embodiment 217. The oligomeric agent of any of embodiments 155-216, wherein each nucleoside of W1 is a 2 ’-MOE nucleoside.

[0385] Embodiment 218. The oligomeric agent of any of embodiments 155-217, wherein W1 consists of four contiguous 2 ’-MOE nucleosides.

[0386] Embodiment 219. The oligomeric agent of any of embodiments 155-218, wherein each 2’- deoxynucleoside of W1 is a 2’-P-D-deoxyribosyl nucleoside.

[0387] Embodiment 220. The oligomeric agent of any of embodiments 155-219, wherein each bicyclic nucleoside of W1 is an LNA nucleoside.

[0388] Embodiment 221. The oligomeric agent of any of embodiments 155-220, wherein W2 comprises 1- 5 modified nucleosides comprising a modified furanosyl sugar moiety.

[0389] Embodiment 222. The oligomeric agent of any of embodiments 155-221, wherein W2 comprises 3 modified nucleosides comprising a modified furanosyl sugar moiety.

[0390] Embodiment 223. The oligomeric agent of any of embodiments 155-221, wherein W2 comprises 5 modified nucleosides comprising a modified furanosyl sugar moiety. CHEMO 113W0

[0391] Embodiment 224. The oligomeric agent of any of embodiments 155-222, wherein W2 consists of 3, 4, 5, or 6 linked nucleosides.

[0392] Embodiment 225. The oligomeric agent of any of embodiments 155-222, wherein W2 consists of 3, 4, or 5 linked nucleosides.

[0393] Embodiment 226. The oligomeric agent of any of embodiments 155-225, wherein each nonterminal modified nucleoside of W2 is linked at its 3’-position by a phosphodiester or a phosphorothioate intemucleoside linkage, optionally a phosphorothioate intemucleoside linkage.

[0394] Embodiment 227. The oligomeric agent of any of embodiments 155-226, wherein W2 does not comprise a 2 ’-deoxynucleoside.

[0395] Embodiment 228. The oligomeric agent of any of embodiments 155-226, wherein W2 comprises at least one 2 ’-deoxynucleoside.

[0396] Embodiment 229. The oligomeric agent of any of embodiments 155-228, wherein each modified nucleoside of W2 is independently selected from a non-bicyclic 2 ’-substituted nucleoside and a bicyclic nucleoside.

[0397] Embodiment 230. The oligomeric agent of any of embodiments 155-228, wherein each modified nucleoside of W2 is independently selected from a non-bicyclic 2 ’-substituted nucleoside and a bicyclic nucleoside other than a cEt nucleoside.

[0398] Embodiment 231. The oligomeric agent of any of embodiments 155-230, wherein each modified nucleoside ofW2 is independently a bicyclic nucleoside.

[0399] Embodiment 232. The oligomeric agent of any of embodiments 155-231, wherein each bicyclic nucleoside of W2 is selected from cEt and LNA.

[0400] Embodiment 233. The oligomeric agent of any of embodiments 155-228, wherein each modified nucleoside ofW2 is independently a non-bicyclic 2 ’-substituted nucleoside.

[0401] Embodiment 234. The oligomeric agent of any of embodiments 155-233, wherein each 2’- substituted nucleoside of W2 is a 2 ’-MOE nucleoside.

[0402] Embodiment 235. The oligomeric agent of any of embodiments 155-233, wherein each 2’- deoxynucleoside of W2 is a 2’-P-D-deoxyribosyl nucleoside.

[0403] Embodiment 236. The oligomeric agent of any of embodiments 155-235, wherein each nucleoside of G is a 2’-p-D-deoxyribosyl nucleoside.

[0404] Embodiment 237. The oligomeric agent of any of embodiments 155-235, wherein each nucleoside of G is linked at its 3 ’-position by a phosphorothioate or a mesyl phosphoramidate intemucleoside linkage. CHEMO 113W0

[0405] Embodiment 238. The oligomeric agent of any of embodiments 155-237, wherein at least one, optionally exactly one, nucleoside of G is linked at its 3 ’-position by a mesyl phosphoramidate intemucleoside linkage.

[0406] Embodiment 239. The oligomeric agent of any of embodiments 155-238, wherein exactly two, exactly three, exactly four, exactly five, or exactly six nucleosides of G are linked at their respective 3’-positions by a mesyl phosphoramidate intemucleoside linkage.

[0407] Embodiment 240. The oligomeric agent of any of embodiments 155-238, wherein exactly five, exactly six, exactly seven, exactly eight, exactly nine, or exactly ten nucleosides of G are linked at their respective 3 ’-positions by a phosphorothioate intemucleoside linkage.

[0408] Embodiment 241. The oligomeric agent of any of embodiments 155-238, wherein the 3’-most nucleoside of G is linked at its 3 ’-position by a mesyl phosphoramidate or phosphorothioate intemucleoside linkage.

[0409] Embodiment 242. The oligomeric agent of any of embodiments 155-167, 169-199, or 202-241, wherein G consists of 7-15 linked nucleosides, optionally 10-13 linked nucleosides, optionally exactly 10 linked nucleosides.

[0410] Embodiment 243. The oligomeric agent of any of embodiments 155-242, wherein G is a deoxy region.

[0411] Embodiment 244. The oligomeric agent of any of embodiments 155-243, wherein W2 has the formula -(E)n4-(D4)n5-(F)n6 wherein each F is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’- deoxy nucleoside, provided that at least one F is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; and each non-terminal 2 ’-deoxy nucleoside of F is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; each E is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety and a sugar surrogate nucleoside; each D4 is independently a 2’-deoxy nucleoside provided that each D4 is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; n6 is 1 to 8 and each of n4 and n5 is independently 0 to 8, provided that n4 + n5 + n6 is less than or equal to 8

[0412] Embodiment 245. The oligomeric agent of embodiment 244, wherein n4 is 1 or 2.

[0413] Embodiment 246. The oligomeric agent of embodiment 244 or 245, wherein n5 is 1.

[0414] Embodiment 247. The oligomeric agent of embodiment 244 or 245, wherein n5 is 0. Embodiment 248. The oligomeric agent of any one of embodiments 244-247, wherein n6 is 1 to 5.

[0415] Embodiment 249. The oligomeric agent of any one of embodiments 244-248, wherein n6 is 1, 2, or 3.

[0416] Embodiment 250. The oligomeric agent of any one of embodiments 244-249, wherein each E is selected from a bicyclic nucleoside and a 2 ’-substituted nucleoside.

[0417] Embodiment 251. The oligomeric agent of any one of embodiments 244-250, wherein each E is a bicyclic nucleoside.

[0418] Embodiment 252. The oligomeric agent of any one of embodiments 244-251, wherein each bicyclic nucleoside of E is selected from cEt and LNA, optionally cEt.

[0419] Embodiment 253. The oligomeric agent of any one of embodiments 244-250, wherein each E is a non-bicyclic 2 ’-substituted nucleoside.

[0420] Embodiment 254. The oligomeric agent of any one of embodiments 244-253, wherein each 2’- substituted nucleoside of E is a 2’-MOE nucleoside.

[0421] Embodiment 255. The oligomeric agent of any one of embodiments 244-254, wherein each D4 is a 2’-P-D-deoxyribosyl nucleoside.

[0422] Embodiment 256. The oligomeric agent of any one of embodiments 244-255, wherein each F is selected from a bicyclic nucleoside and a non-bicyclic 2 ’-substituted nucleoside.

[0423] Embodiment 257. The oligomeric agent of any one of embodiments 244-256, wherein each F is a non-bicyclic 2 ’-substituted nucleoside.

[0424] Embodiment 258. The oligomeric agent of any one of embodiments 244-256, wherein non-bicyclic 2 ’-substituted nucleoside of F is a 2’-MOE nucleoside.

[0425] Embodiment 259. The oligomeric agent of any one of embodiments 244-256 or 258, wherein each F is a bicyclic nucleoside.

[0426] Embodiment 260. The oligomeric agent of any one of embodiments 244-259, wherein each bicyclic nucleoside of F is selected from cEt and LNA, optionally cEt.

[0427] Embodiment 261. The oligomeric agent of any one of embodiments 244-260, wherein each of n4 and n5 is 0.

[0428] Embodiment 262. The oligomeric agent of any one of embodiments 244-261, wherein n6 is 5, and each of n4 and n5 is 0.

[0429] Embodiment 263. The oligomeric agent of any of embodiments 155-262, wherein W1 has the formula (A)nl-(Dl)n2-(B)n3- wherein each A is independently selected from a modified CHEMO 113W0 nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’- deoxy nucleoside provided that at least one A is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; and each 2 ’-deoxy nucleoside of A is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; each B is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety and a sugar surrogate nucleoside; each DI is independently a 2’-deoxy nucleoside provided that each DI is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; nl is 1 to 8 and each of n2 and n3 is independently 0 to 8, provided that nl + n2 + n3 is less than or equal to 8

[0430] Embodiment 264. The oligomeric agent of embodiment 263, wherein n3 is 0 or 1.

[0431] Embodiment 265. The oligomeric agent of embodiment 263 or 264, wherein n2 is 0 or 1.

[0432] Embodiment 266. The oligomeric agent of any one of embodiments 263-265, wherein nl is 1 to 5.

[0433] Embodiment 267. The oligomeric agent of any one of embodiments 263-265, wherein nl is 2 or 3.

[0434] Embodiment 268. The oligomeric agent of any one of embodiments 263-267, wherein each B is selected from a bicyclic nucleoside and a non-bicyclic 2 ’-substituted nucleoside.

[0435] Embodiment 269. The oligomeric agent of any one of embodiments 263-268, wherein each B is a non-bicyclic 2 ’-substituted nucleoside.

[0436] Embodiment 270. The oligomeric agent of any one of embodiments 263-269, wherein each non- bicyclic 2 ’-substituted nucleoside of B is a 2’-MOE nucleoside.

[0437] Embodiment 271. The oligomeric agent of any one of embodiments 263-270, wherein each B is a bicyclic nucleoside.

[0438] Embodiment 272. The oligomeric agent of any one of embodiments 263-271, wherein each bicyclic nucleoside of B is selected from cEt and LNA, optionally cEt.

[0439] Embodiment 273. The oligomeric agent of any one of embodiments 263-272, wherein each D4 is a 2’-P-D-deoxyribosyl nucleoside.

[0440] Embodiment 274. The oligomeric agent of any one of embodiments 263-273, wherein each A is selected from a bicyclic nucleoside and a 2 ’-substituted nucleoside.

[0441] Embodiment 275. The oligomeric agent of any one of embodiments 263-274, wherein each A is a 2’ -substituted nucleoside. CHEMO 113W0

[0442] Embodiment 276. The oligomeric agent of any one of embodiments 263-275, wherein each 2’- substituted nucleoside of A is a 2 ’-MOE nucleoside.

[0443] Embodiment 277. The oligomeric agent of any one of embodiments 263-274 or 276, wherein each A is a bicyclic nucleoside.

[0444] Embodiment 278. The oligomeric agent of any one of embodiments 263-277, wherein each bicyclic nucleoside of A is selected from cEt and LNA, optionally cEt.

[0445] Embodiment 279. The oligomeric agent of any one of embodiments 263-278, wherein each of n2 and n3 is 0.

[0446] Embodiment 280. The oligomeric agent of any one of embodiments 263-279, wherein nl is 4, and each of n2 and n3 is 0.

[0447] Embodiment 281. The oligomeric agent of any of embodiments 263-279, wherein the modified oligonucleotide consists of 17-20 linked nucleosides.

[0448] Embodiment 282. The oligomeric agent of any of embodiments 263-281, wherein the modified oligonucleotide consists of 17, 18, 19, or 20 linked nucleosides.

[0449] Embodiment 283. The oligomeric agent of any of embodiments 263-280, wherein the modified oligonucleotide consists of 20 linked nucleosides.

[0450] Embodiment 284. The oligomeric agent of any one of embodiments 263-200 or 201-283, wherein G consists of 11-15 linked nucleosides, optionally 11, optionally 12, optionally 13, optionally 14, or optionally 15 linked nucleosides, optionally wherein neither W1 nor W2 comprises a 2’- deoxynucleoside.

[0451] Embodiment 285. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0452] Ak-Wl-G-W2

[0453] I wherein Akis a cEt adenosine, W1 consists of 4 linked nucleosides, wherein each nucleoside of W1 is selected from a non-bicyclic 2 ’-substituted nucleoside and a bicyclic nucleoside;

[0454] G consists of 10 linked 2'-P-D-deoxyribosyl nucleosides;

[0455] W2 consists of 5 linked nucleosides, wherein each nucleoside of W2 is independently selected from a non-bicyclic 2 ’-substituted nucleoside and a bicyclic nucleoside; CHEMO 113W0 provided that the modified oligonucleotide comprises at least one intemucleoside linkage other than a phosphorothioate or phosphodiester intemucleoside linkage; optionally wherein the modified oligonucleotide consists of the region of formula I

[0456] Embodiment 286. The oligomeric agent of embodiment 285, wherein the modified oligonucleotide comprises at least one phosphodiester intemucleoside linkage.

[0457] Embodiment 287. The oligomeric agent of embodiment 285, wherein the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to an Lpa transcript (SEQ ID NO: 1).

[0458] Embodiment 288. The oligomeric agent of any of embodiments 285-287, wherein W1 comprises exactly one bicyclic nucleoside.

[0459] Embodiment 289. The oligomeric agent of any of embodiments 285-287, wherein each nucleoside of W1 is a non-bicyclic 2 ’-substituted nucleoside.

[0460] Embodiment 290. The oligomeric agent of any one of embodiments 285-289, wherein each non- bicyclic 2 ’-substituted nucleoside ofWl is selected from a 2’-MOE nucleoside and a 2’-OMe nucleoside.

[0461] Embodiment 291. The oligomeric agent of embodiment 290, wherein each non-bicyclic 2’- substituted nucleoside of W1 is a 2 ’-MOE nucleoside.

[0462] Embodiment 292. The oligomeric agent of any one of embodiments 285-291, wherein each bicyclic nucleoside of W1 is selected from a cEt nucleoside and an LNA nucleoside.

[0463] Embodiment 293. The oligomeric agent of any one of embodiments 285-291, wherein each bicyclic nucleoside of W1 is a cEt nucleoside.

[0464] Embodiment 294. The oligomeric agent of any one of embodiments 285-293, wherein W2 comprises no bicyclic nucleosides.

[0465] Embodiment 295. The oligomeric agent of any one of embodiments 285-294, wherein W2 comprises exactly one bicyclic nucleoside.

[0466] Embodiment 296. The oligomeric agent of any one of embodiments 285-294, wherein W2 comprises exactly two bicyclic nucleosides.

[0467] Embodiment 297. The oligomeric agent of any one of embodiments 285-296, wherein each non- bicyclic 2 ’-substituted nucleoside of W2 is selected from a 2’-MOE nucleoside and a 2’-OMe nucleoside.

[0468] Embodiment 298. The oligomeric agent of embodiment 285, wherein each non-bicyclic 2’- substituted nucleoside of W2 is a 2 ’-MOE nucleoside. CHEMO 113W0

[0469] Embodiment 299. The oligomeric agent of any one of embodiments 285-298, wherein each intemucleoside linkage is selected from a phosphodiester, a phosphorothioate, and a mesyl phosphoramidate .

[0470] Embodiment 300. The oligomeric agent of any of embodiments 155-299, wherein the modified oligonucleotide comprises nucleobases independently selected from thymine, uracil, guanine, cytosine, 5 -methylcytosine, adenine, and hypoxanthine.

[0471] Embodiment 301. The oligomeric agent of any of embodiments 155-300, comprising a conjugate group.

[0472] Embodiment 302. The oligomeric agent of embodiment 301, wherein the conjugate group comprises a carbohydrate or carbohydrate cluster.

[0473] Embodiment 303. The oligomeric agent of embodiment 301or 302, wherein the conjugate group comprises at least one GalNAc.

[0474] Embodiment 304. The oligomeric agent of embodiment 301, wherein the conjugate group comprises a C10-C20 alkyl chain.

[0475] Embodiment 305. The oligomeric agent of embodiment 304, wherein the conjugate group comprises Cie alkyl.

[0476] Embodiment 306. The oligomeric agent of embodiment 301, wherein the conjugate group comprises a transferrin receptor ligand.

[0477] Embodiment 307. The oligomeric agent of embodiment 306, wherein the transferrin receptor ligand comprises a peptide.

[0478] Embodiment 308. The oligomeric agent of embodiment 306 or 307, wherein the transferrin receptor ligand comprises a bicyclic peptide.

[0479] Embodiment 309. The oligomeric agent of embodiment 306, wherein the transferrin receptor ligand comprises a Fab, Fab’, F(ab’)2, scFv, VHH, VNAR, or nanobody.

[0480] Embodiment 310. The oligomeric agent of any one of embodiments 301-309, wherein the oligomeric agent comprises the modified oligonucleotide, the conjugate group, and a cleavable linker comprising nucleosides linked by phosphodiester linkages linking the modified oligonucleotide and the conjugate group.

[0481] Embodiment 311. The oligomeric agent of any one of embodiments 155-300, wherein the oligomeric agent consists of the modified oligonucleotide.

[0482] Embodiment 312. The oligomeric agent of any of embodiments 155-311, wherein the nucleobase sequence of the modified oligonucleotide is complementary to a target nucleic acid. CHEMO 113W0

[0483] Embodiment 313. The oligomeric agent of embodiment 312, wherein the nucleobase sequence of the modified oligonucleotide is at least 80%, at least 90%, or 100% complementary to the target nucleic acid.

[0484] Embodiment 314. The oligomeric agent of embodiment 312 or 313, wherein the target nucleic acid is selected from: an mRNA, a pre-mRNA, a microRNA, and a non-coding RNA.

[0485] Embodiment 315. The oligomeric agent of embodiment 312, wherein the target nucleic acid is not a

[0486] CNS target.

[0487] Embodiment 316. The oligomeric agent of embodiment 312, wherein the target nucleic acid is a

[0488] CNS target.

[0489] Embodiment 317. The oligomeric agent of any of embodiments 155-316, wherein the modified oligonucleotide is a first modified oligonucleotide, and further comprising a second modified oligonucleotide that is complementary to the first modified oligonucleotide.

[0490] Embodiment 318. A pharmaceutical composition comprising the oligomeric agent of any of embodiments 155-317 and a pharmaceutically acceptable carrier.

[0491] Embodiment 319. A method comprising contacting a cell with the oligomeric agent or pharmaceutical composition of any of embodiments 155-317.

[0492] Embodiment 320. A method of modulating the amount or activity of a target nucleic acid in a cell, comprising contacting the cell with the oligomeric agent of any of embodiments 155-317 or pharmaceutical composition of embodiment 318.

[0493] Embodiment 321. The method of embodiment 319-320, whereby the amount or activity of a target nucleic acid is reduced.

[0494] Embodiment 322. The method of any one of embodiments 319-321, comprising parenteral administration to a subject.

[0495] Embodiment 323. The method of any one of embodiments 319-322, comprising injection into CSF.

[0496] Embodiment 324. The method of any one of embodiments 319-322, comprising subcutaneous injection.

[0497] Embodiment 325. The oligomeric agent of any of embodiments 155-317 or the pharmaceutical composition of embodiment 318, for use in treatment of a disease or condition.

[0498] Embodiment 326. The oligomeric agent of any of embodiments 155-317 or the pharmaceutical composition of embodiment 318, for use in the manufacture of a medicament.

[0499] Embodiment 327. The oligomeric agent of any of embodiments 155-317 or the pharmaceutical composition of embodiment 318, for use in therapy. CHEMO 113W0

[0500] Oligomeric Agents

[0501] Provided herein are oligomeric agents comprising or consisting of at least one modified oligonucleotide and optionally one or more additional associated features selected from: (a) one or more additional modified or unmodified oligonucleotides, each of which may be hybridized to or covalently linked to the at least one modified oligonucleotide and / or to each other; (b) one or more conjugate groups, which may be covalently attached to any oligonucleotide of such oligomeric agent; and (c) one or more terminal groups. In some embodiments, provided herein are oligomeric agents comprising or consisting of a modified antisense oligonucleotide complementary to target RNA. In certain such embodiments, an oligomeric agent consists of a modified antisense oligonucleotide complementary to target RNA and a conjugate group attached to the modified antisense oligonucleotide. In some embodiments, provided herein are oligomeric agents comprising an oligomeric duplex comprising or consisting of an antisense oligonucleotide complementary to target RNA, and a sense oligonucleotide complementary to the antisense oligonucleotide, wherein one or both of the antisense and sense oligonucleotides is / are modified. In certain such embodiments, an oligomeric agent consists of an antisense oligonucleotide complementary to target RNA, a sense oligonucleotide complementary to the antisense oligonucleotide, and a conjugate group and / or terminal group attached to one or both of the antisense and sense oligonucleotides. Modified antisense and / or sense oligonucleotides comprise at least one modified nucleoside (comprising a modified sugar moiety and / or a modified nucleobase and / or lacking a nucleobase) and / or at least one modified intemucleoside linkage. Examples of certain modified nucleosides and modified intemucleoside linkages suitable for use in modified antisense and / or sense oligonucleotides are described herein.

[0502] A. Modified Nucleosides

[0503] In certain embodiments, modified nucleosides comprising the following modified sugar moieties and / or the following modified nucleobases may be incorporated into oligonucleotides described herein.

[0504] / . Modified Sugar Moie ties

[0505] In certain embodiments, a modified sugar moiety is a 2 ’-substituted sugar moiety. In certain embodiments, a modified sugar moiety is a bicyclic sugar moiety.

[0506] In certain embodiments, modified sugar moieties are non-bicyclic modified furanosyl sugar moieties optionally comprising one or more substituent groups including, but not limited to, substituents at the 2', 3', 4', and / or 5' positions, as numbered below: CHEMO 113W0

[0507] In certain embodiments, a non-bicyclic modified sugar moiety is a 2'-substituted sugar moiety which comprises a substituent group at the 2'-position. Examples of substituent groups suitable for the 2'- position of modified sugar moieties include but are not limited to: F, OCH3 (“OMe” or “O-methyl”), and O(CH2)2OCHS (“MOE” or “O-methoxyethyl” or OCH2CH2OCH3). In certain embodiments, 2'-substituent groups are selected from: halo, allyl, amino, azido, SH, CN, OCN, CF3, OCF3, C1-C10 alkoxy, substituted C1-C10 alkoxy, C1-C10 alkyl, substituted C1-C10 alkyl, S-alkyl, N(Rm)-alkyl, O-alkenyl, S-alkenyl, N(Rm)- alkenyl, O-alkynyl, S-alkynyl, N(Rm)-alkynyl, O-alkylenyl-O-alkyl, alkynyl, alkaryl, aralkyl, O-alkaryl, O- aralkyl, O(CH2)2SCH3, O(CH2)2ON(Rm)(Rn) or 0CH2C(=0)-N(Rm)(Rn), where each Rmand Rnis, independently, H, an amino protecting group, or substituted or unsubstituted C1-C10 alkyl, O(CH2)2ON(CH3)2 (“DMAOE”), or O(CH2)2O(CH2)2N(CH3)2 (“DMAEOE”). Synthetic methods for some of these 2'-substituent groups may be found, e.g., in Cook et al., U.S. 6,531,584; Cook et al., U.S. 5,859,221; and Cook et al., U.S. 6,005,087. Certain embodiments of these 2 '-substituent groups may be further substituted with one or more substituent groups independently selected from: halo, cyano, ORa2, NO2, NH2, NHRa2, N(Ra2)2, Ci-Ce alkyl, Ci-Ce haloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C10 cycloalkyl, Ce-Cio aryl, heteroaryl, heterocyclyl, Ci-Ce alkylcnc-NIU. Ci-Ce alkylene-NHRa2, Ci-Ce alkylene-N(Ra2)2, C(O)Ra3, C(O)ORa3, C(O)NHRa3, C(O)N(Ci-C4alkyl)Ra3, SRa3, S(O)2Ra3, S(O)Ra3, NHC(O)Ra3, N(CI-C4alkyl)C(O)Ra3, NHS(O)Ra3, N(Ci-C4alkyl)S(O)Ra3, NHS(O)2Ra3, and N(Ci-C4alkyl)S(O)2Ra3; where each Ra2is independently selected from C2-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C10 cycloalkyl, Ce-Cioaryl, heteroaryl, and heterocyclyl; and each Ra3is independently hydrogen, OH, Ci-Ce alkyl, Ci-Cehaloalkyl, C3- C10 cycloalkyl, Ce-Cio aryl, heteroaryl, or heterocyclyl. In certain embodiments, a sugar moiety comprises two of the above substituents at the 2'-position. In certain embodiments, a sugar moiety comprises a 2'- fluoro and a second 2'-substituent.

[0508] In certain embodiments, a 2'-substituted sugar moiety comprises a non-bridging 2'-substituent group selected from: F, NH2, N3, OCF3, OCH3, O(CH2)3NH2, CH2CH=CH2, OCH2CH=CH2, OCH2CH2OCH3, O(CH2)2SCH3, O(CH2)2ON(Rm)(Rn), O(CH2)2O(CH2)2N(CH3)2, and N-substituted acetamide (OCH2C(=O)-N(Rm)(Rn)), where each Rmand Rnis, independently, H, an amino protecting group, or substituted or unsubstituted C1-C10 alkyl. In certain embodiments, a 2'-substituted sugar moiety comprises a non-bridging 2'-substituent group selected from: F, OCF3OCH3, OCH2CH2OCH3, O(CH2)2SCH3, O(CH2)2ON(CH3)2, O(CH2)2O(CH2)2N(CH3)2, O(CH2)2ON(CH3)2 (“DMAOE”), O(CH2)2O(CH2)2N(CH3)2 (“DMAEOE”), and OCH2C(=O)-N(H)CH3(“NMA”). In certain embodiments one or more non-bridging substituent of non-bicyclic modified sugar moieties is branched.

[0509] In certain embodiments, a 2'-substituted sugar moiety comprises a 2'-substituent group selected from: F, OCH3, and O(CH2)2OCH3. In certain embodiments, a 2'-substituted sugar moiety comprises a 2'- substituent group selected from: OCH3, O(CH2)2OCH3, and OCH2C(=O)-N(H)CH3.

[0510] In certain embodiments, furanosyl sugar moieties and nucleosides incorporating such furanosyl sugar moieties are further defined by stereochemical configuration. For example, a 2'-deoxyfuranosyl sugar moiety (i.e., 2'-(H)H furanosyl sugar moiety) may be in seven isomeric configurations other than the CHEMO 113W0 naturally occurring P-D-deoxyribosyl configuration. Such modified sugar moieties are described in, e.g., WO 2020 / 072991. A 2'-modified sugar moiety has an additional stereocenter at the 2'-position relative to a 2'-deoxyfuranosyl sugar moiety; therefore, such sugar moieties have a total of sixteen possible stereochemical configurations. Modified fiiranosyl sugar moieties described herein are in the -D-ribosyl stereochemical configuration unless otherwise specified.

[0511] In certain embodiments, the sugar moiety is a xylosyl, lyxosyl, or arabinosyl sugar moiety.

[0512] In certain embodiments, non-bicyclic modified sugar moieties comprise a substituent group at the 4'-position. Examples of substituent groups suitable for the 4 '-position of modified sugar moieties include, but are not limited to, alkoxy (e.g., methoxy), alkyl, and those described in Manoharan et al., WO 2015 / 106128.

[0513] In certain embodiments, non-bicyclic modified sugar moieties comprise a substituent group at the 3 '-position. Examples of substituent groups suitable for the 3 '-position of modified sugar moieties include, but are not limited to, alkoxy (e.g., methoxy), alkyl (e.g., methyl, ethyl).

[0514] In certain embodiments, non-bicyclic modified sugar moieties comprise a substituent group at the 5 '-position. Examples of substituent groups suitable for the 5 '-position of modified sugar moieties include, but are not limited to, vinyl, alkoxy (e.g. , methoxy), alkynyl, allyl, and alkyl (e.g. , methyl (R or .S). ethyl (R or S)).

[0515] In certain embodiments, non-bicyclic modified sugar moieties comprise more than one nonbridging sugar substituent, for example, 2'-F-5'-methyl sugar moieties, such as described in Migawa et al., US 2010 / 0190837, or alternative 2'- and 5'-modified sugar moieties as described in Rajeev et al., US 2013 / 0203836.

[0516] Certain modified sugar moieties are bicyclic sugar moieties, which comprise a substituent that bridges two atoms of the fiiranosyl ring to form a second ring. In certain embodiments, the bicyclic sugar moiety comprises a bridge between the 4' and the 2' furanose ring atoms. Examples of such 4' to 2' bridging sugar substituents include, but are not limited to: 4'-CH2-2', 4'-(CH2)2-2', 4'-(CH2)3-2', 4'-CH2-O-2' (“LNA”), 4'-CH2-S-2', 4'-(CH2)2-O-2' (“ENA”), 4'-CH(CH3)-O-2' (referred to as “constrained ethyl” or “cEt” when in the S configuration), 4'-CH2-O-CH2-2', 4'-CH2-N(R)-2', 4'-CH(CH2OCH3)-O-2' (“constrained MOE” or “cMOE”) and analogs thereof, 4'-C(CH3)(CH3)-O-2' and analogs thereof, 4'-CH2- N(OCH3)-2' and analogs thereof, 4'-CH2-O-N(CH3)-2', 4'-CH2-C(H)(CH3)-2', 4'-CH2-C(=CH2)-2' and analogs thereof, 4'-C(RaRb)-N(R)-O-2', 4'-C(RaRb)-O-N(R)-2', 4'-CH2-O-N(R)-2', and 4'-CH2-N(R)-O-2', wherein each R, Ra, and Rb is, independently, H, a protecting group, or C1-C12 alkyl. Representative U.S. patents that teach the preparation of such bicyclic sugar moieties include, but are not limited to: hnanishi et al., U.S. 7,427,672; Swayze et al., U.S. 7,741,457; Swayze et al., U.S. 8,022,193; Seth et al., U.S. 8,278,283; Prakash et al., U.S. 8,278,425; and Seth et al., U.S. 8,278,426.

[0517] In certain embodiments, such 4' to 2' bridges independently comprise from 1 to 4 linked groups independently selected from: -[C(Ra)(Rb)]n-, -[C(Ra)(Rb)]n-O-, -C(Ra)=C(Rb)-, -C(Ra)=N-, -C(=NRa)-, -

[0518] C(=O)-, -C(=S)-, -O-, -Si(Ra)2-, -S(=O)x-, and -N(Ra)-; wherein: CHEMO 113W0 x is 0, 1, or 2; n is 1, 2, 3, or 4; each Ra and Rb is, independently, halo, cyano, ORa2, NO2, NH2, NHRa2, N(Ra2)2, Ci-Ce alkyl, Ci- Ce haloalkyl, C2-Ce alkenyl, C2-Ce alkynyl, C3-C10 cycloalkyl, Ce-io aryl, heteroaryl, heterocyclyl, Ci-Ce alkylene-NH2, Ci-C6alkylene-NHRa2, Ci-C6alkylene -N(Ra2)2, C(O)Ra3, C(O)ORa3, C(O)NHRa3, C(O)N(Ci-C4alkyl)Ra3, SRa3, S(O)2Ra3, S(O)Ra3, NHC(O)Ra3, N(Ci-C4alkyl)C(O)Ra3, NHS(O)Ra3, N(Ci- C4alkyl) S(O)Ra3, NHS(O)2Ra3, and N(Ci-C4alkyl)S(O)2Ra3; each Ra2is independently selected from Ci-Ce alkyl, C2-Ce alkenyl, C2-Ce alkynyl, C3-C10 cycloalkyl, Ce-Cioaryl, heteroaryl, and heterocyclyl; each Ra3is independently hydrogen, OH, Ci-Ce alkyl, Ci-Ce haloalkyl, C3-C10 cycloalkyl, Ce-Cio aryl, heteroaryl, or heterocyclyl.

[0519] In certain embodiments, the bicyclic sugar moiety comprises a bridge between the 5' and the 3' furanose ring atoms. Examples of such 5' to 3' bridging sugar substituents include, but are not limited to, 5'-(CH2)2-3' (bcDNA), 5'-(CH2)3-3' (bc43DNA), 5'-C(F)=CH-CH2-3', and 5'-CH2-CHQ-3', wherein Q is an attachment to an intemucleoside linkage.

[0520] Additional bicyclic sugar moieties are known in the art, see, for example: Wan, et al., J. Medicinal Chemistry, 2016, 59, 9645-9667; Wengel et al., U.S. 8,080,644; Ramasamy et al., U.S. 6,525,191; Seth et al., U.S. 7,547,684; and Seth et al., U.S. 7,666,854.

[0521] In certain embodiments, bicyclic sugar moieties and nucleosides incorporating such bicyclic sugar moieties are further defined by stereochemical configuration. For example, an LNA nucleoside (described herein) may be in the a-L configuration or in the -D configuration.

[0522] LNA (p-D-configuration) LNA (a-L-configuration) bridge = 4’-CH2-O-2’ bridge = 4'-CH2-O-2' a-L-methyleneoxy (4'-CH2-O-2') or a-L-LNA bicyclic nucleosides have been incorporated into oligonucleotides that showed antisense activity (e.g., Frieden et al. Nucleic Acids Res. 2003, 21, 6365- 6372). The addition of locked nucleic acids to siRNAs has been shown, in certain studies, to increase siRNA stability in serum, and to reduce off-target effects (Elmen, J. et al. Nucleic Acids Res. 2005, 33(1), 439-447; Mook, O. R. etal. Mol. Cane. Ther. 2007, 6(3), 833-843; Grunweller, A. etal. Nucleic Acids Res. 2003, 37(12), 3185-3193). Herein, general descriptions of bicyclic nucleosides include both stereochemical configurations. When the positions of specific bicyclic nucleosides (e.g., LNA or cEt) are identified in exemplified embodiments herein, they are in the P-D stereochemical configuration, unless otherwise specified.

[0523] In certain embodiments, a bicyclic sugar moiety may comprise a substituent in addition to the bridging substituent (e.g., a sugar moiety comprising a 5 '-substituent and a 4'-2' bridging substituent). CHEMO 113W0

[0524] In certain embodiments, a nucleoside is a sugar surrogate nucleoside. In certain embodiments, the sugar surrogate is selected from a cyclic sugar surrogate and an acyclic sugar surrogate.

[0525] A cyclic sugar surrogate is represented by Formula X:

[0526] Formula X

[0527] Wherein:

[0528] J is H, Ci-Ce alkyl, or C2-C6 alkenyl;

[0529] X is O, S, C(R1R2), N(R3), or X1-X2, wherein X1-X2 is C(RI)=C(R2), C(RIR2)-C(RIR2), O- C(RIR2), C(RIR2)-O, S-C(RIR2), C(RIR2)-S, N(R3)-C(RIR2), or C(RIR2)-N(R3);

[0530] Y is C(RIR2) or YI.Y2, wherein YJ.Y2is C(RI)=C(R2), or C(RIR2)-C(RIR2);

[0531] Z is C(GIG2) or Zi-Z2, wherein Z1-Z2 is C(Gi)=C(Ri), C(Ri)=C(Gi), C(GIG2)-C(RIR2), C(RIR2)- C(GIG2), C(GIG2)-C(RIR2)-C(RIR2), or C(RIR2)-C(RIR2)-C(GIG2);

[0532] Q is CH or N; each Ri and R2is independently H, OH, Ci-Ce alkyl, or N(R0; wherein if Ri is OH, then R2is not OH; each R3and R4 is independently H, Ci-Ce alkyl, or C(=O)R5, wherein R5 is Ci-Ce alkyl; each Gi and G2is independently H, OH, halogen or O- 1 C ( R ,)( R?) | -| (C =O) -X | -R^i wherein if

[0533] Gi is OH, then G2 is not OH; each Re and R7 is, independently, H, halogen, Ci-Ce alkyl or substituted Ci-Ce alkyl; each XGis O, S orN(Ei);

[0534] Rs is H, halogen, Ci-Ce alkyl, substituted Ci-Ce alkyl, C2-C6 alkenyl, substituted C2-C6 alkenyl, C2-C6 alkynyl, substituted C2-C6 alkynyl or N(E2)(E3);

[0535] Ei, E2and E3are each, independently, H, Ci-Ce alkyl or substituted Ci-Ce alkyl; m is 0 or 1; p is 0 or 1; q is from 1 to 6; s is 0 or 1; j is 0 or 1; and with the proviso that if X is O, Z is C(GIG2), and Q is CH, then m is 1.

[0536] In certain such embodiments, the oxygen atom of the sugar moiety is replaced, e.g., with a sulfur, carbon or nitrogen atom (X is S, C(Ri R2), or N(R3)). In certain such embodiments, such modified sugar moieties also comprise bridging and / or non-bridging substituents as described herein. For example, certain cyclic sugar surrogates comprise a 4'-sulfur atom and a substitution at the 2'-position and / or the 5' position. CHEMO 113W0

[0537] In certain embodiments, cyclic sugar surrogates comprise rings having other than 5 atoms. For example, in certain embodiments, a cyclic sugar surrogate comprises a six-membered tetrahydropyran (“THP”), where X in Formula la is 0-C(RIR2), p is 1, Q is CH, Z is C(GIG2), and m is 0. Such tetrahydropyrans may be further modified or substituted. Nucleosides comprising such modified tetrahydropyrans include but are not limited to hexitol nucleic acid (“HNA”), altritol nucleic acid (Gi=OH; G2=H; “ANA”), and fluoro HNA (FHNA):

[0538] FHNA

[0539] (Gi=F; G2=H; “FHNA”, see e.g., Egli, M. et al. J. Am. Chem. Soc. 2011, 733(41), 16642-16649; Swayze et al., U.S. 8,088,904; and Swayze et al., U.S. 8,440,803); FHNA can also be referred to as a F-THP or 3 '-fluoro tetrahydropyran or 3 '-FHNA).

[0540] In certain embodiments, cyclic sugar surrogates comprise rings having more than 5 atoms and more than one heteroatom. For example, nucleosides comprising morpholino sugar moieties and their use in oligonucleotides have been reported. As used here, the term “morpholino” means a sugar surrogate having Formula la, above, wherein X is O, Y and Z are each CH2, and Q is N.

[0541] In certain embodiments, a morpholino is modified, for example by adding or altering various substituent groups from the above morpholino structure. Such sugar surrogates are referred to herein as “modified morpholines. ”

[0542] An acyclic sugar surrogate nucleoside means a nucleoside having Formula XI, Formula XII, or Formula Xlla: CHEMO 113W0

[0543] Formula Xlla wherein

[0544] X is O, S, C(R5Re), N(Ei), NC(=O)-(Ei); each Ji and J2are independently H or Ci-Ce alkyl; n is 0, 1 or 2; m is 0, 1, or 2; p is 0 or 1; o is 0 or 1; s is 0 or 1;

[0545] Ri is H, OH, halogen, Ci-Ce alkyl, Ci-Ce alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, or (CH2)qRs;

[0546] R2, R3, and R4 are each independently H, OH, halogen, Ci-Ce alkyl, Ci-Ce alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, S-CH3, N(CH3)(CH3), OCH2CH2OCH3, O-alkylamino, or (CH2)qR«;

[0547] Ei is H, Ci-Ce alkyl or substituted Ci-Ce alkyl;

[0548] Rs and Rs are independently H, OH, Ci-Ce alkyl, or N(R7); wherein if Rs is OH, then Re is not OH; R7is H, Ci-Ce alkyl, or C(=O)Rg, wherein R, is Ci-Ce alkyl;

[0549] Rs is OH, halogen, methoxy, ethoxy, azido, C2-C6 alkenyl, or C2-C6 alkynyl, and q is 1, 2, or 3; and

[0550] Bx is a nucleobase.

[0551] As used herein, “acyclic sugar surrogate” means the sugar moiety of an acyclic sugar surrogate nucleoside.

[0552] In certain embodiments, acyclic sugar surrogates are the “unlocked” sugar structure of UNA (“unlocked nucleic acid”) nucleosides. Representative U.S. publications that teach the preparation of UNA include, but are not limited to, U.S. Patent Publication No. 2011 / 0313020.

[0553] In certain embodiments, acyclic sugar surrogates are the glycerol as found in GNA (“glycol nucleic acid”) nucleosides, having Formula Ila wherein n is 1, m and o are 0, s is 1, and J2, R2, and R3 are each H, or the butyl as found in acyclic butyl nucleic acid, having Formula Ila wherein n is 2, m and o are 0, s is 1, and J2, R2, and R3 are each H. In certain embodiments, acyclic sugar surrogates are also known as “C3 spacers” and have Formula Ila wherein n and o are 1; m and s are 0, and Ji, J2, Ri, and R3 are each H. In certain embodiments, GNA is (.S)-GNA,

[0554] Further acyclic sugar surrogates include those described in Manoharan etal., U.S. 10,913,767; US patent publication US 2021 / 0238595; and PCT publication WO 2023 / 109940.

[0555] In certain embodiments, modified oligonucleotides include one or more sugar mimic, in which a group of atoms other than a “furanosyl sugar moiety” or a “sugar surrogate” form the portion of a nucleoside CHEMO 113W0 corresponding to the P-D-ribosyl sugar in RNA. In certain embodiments, a sugar mimic is a portion of the backbone of a peptide nucleic acid, while the remainder of the backbone of the peptide nucleic acid is an intemucleoside linkage. Representative U.S. patents that teach the preparation of PNA compounds include, but are not limited to, U.S. Patent Nos. 5,539,082; 5,714,331; and 5,719,262.

[0556] In certain embodiments, the modified oligonucleotide comprises a modification disclosed in U.S. Patent Nos. 10,233,448 or 11,504,391.

[0557] 2. Modified Nucleobases

[0558] In certain embodiments, a modified oligonucleotide comprises one or more nucleoside comprising an unmodified nucleobase. In certain embodiments, a modified oligonucleotide comprises one or more nucleoside comprising a modified nucleobase. In certain embodiments, a modified oligonucleotide comprises one or more sugar surrogate nucleosides that do not comprise a nucleobase. In certain embodiments, a modified oligonucleotide comprises an abasic nucleoside. In certain embodiments, a modified oligonucleotide does not comprise an abasic nucleoside. In certain embodiments, a modified oligonucleotide comprises one or more inosine nucleosides (z.e., nucleosides comprising a hypoxanthine nucleobase). A universal base is a modified nucleobase that can pair with any one of the five unmodified nucleobases.

[0559] In certain embodiments, at least one purine and / or at least pyrimidine is modified. In certain embodiments, at least one adenine is modified. In certain embodiments, at least one guanine is modified. In certain embodiments, at least one thymine is modified. In certain embodiments, at least one uracil is modified. In certain embodiments, at least one cytosine is modified. In certain embodiments, at least one of the cytosine nucleobases in a modified oligonucleotide is 5 -methylcytosine. In certain embodiments, each of the cytosine nucleobases is a 5 -methylcytosine and all of the other nucleobases of the modified oligonucleotide are unmodified nucleobases. In certain embodiments, one or two of the cytosine nucleobases are 5-methylcytosines and all of the other nucleobases of the modified oligonucleotide are unmodified nucleobases. In certain embodiments, each nucleobase is selected from 5 -methylcytosine, unmodified cytosine, unmodified thymine, unmodified uracil, unmodified adenine, unmodified guanine, and hypoxanthine. In certain embodiments, each nucleobase is selected from 5-methylcytosine, unmodified cytosine, unmodified thymine, unmodified adenine, and unmodified guanine. In certain embodiments, each nucleobase is selected from unmodified cytosine, unmodified thymine, unmodified uracil, unmodified adenine, and unmodified guanine. In certain embodiments, each nucleobase is selected from unmodified cytosine, unmodified thymine, unmodified adenine, and unmodified guanine.

[0560] Unless otherwise indicated, modified adenine has structure (I): CHEMO 113W0

[0561] I wherein: R1Ais absent or H; R2Ais H, Ci-Ce alkyl, substituted Ci-Ce alkyl, Ci-Ce thioalkyl, or substituted Ci-Ce thioalkyl, Ci-Ce alkyloxy, or substituted Ci-Ce alkyloxy; R6Ais H, N(Ra)(Rb), oxo, acetyl, formyl, or O-phenyl; Y7Ais N and R7Ais absent or is Ci-Ce alkyl; or Y7Ais C and R7Ais H, Ci-Ce alkyl, or N(Ra)(Rb); Y8Ais N and R8Ais absent, or Y8Ais C and R8Ais H, a halogen, OH, Ci-Ce alkyl, or substituted Ci-Ce alkyl; Raand Rbare each independently H, Ci-Ce alkyl, substituted Ci-Ce alkyl, Ci-Ce alkenyl, substituted Ci-Ce alkenyl, acetyl, or formyl, or together form a 5-7-membered heterocycle; excluding where Y7Ais N and R7Ais absent; Y8Ais C, R8Ais H, R1Ais absent, R2Ais H, and R6Ais NH2(unmodified adenine).

[0562] Unless otherwise indicated, modified guanine has structure (II):

[0563] II wherein: R2Gis N(Ra)(Rb); R6Gis oxo and R1Gis H, or R6Gis O-Ci-Ce alkyl or S-Ci-Ce alkyl and R1Gis absent; Y7Gis N and R7Gis absent or is Ci-Ce alkyl; or Y7Gis C and R7Gis H, Ci-Ce alkyl, or N(Ra)(Rb); Y8Gis N and R8Gis absent, or Y8Gis C and R8Gis H, a halogen, OH, Ci-Ce alkyl, or substituted Ci-Ce alkyl; Raand Rbare independently H, Ci-Ce alkyl, substituted Ci-Ce alkyl, Ci-Ce alkenyl, substituted Ci-Ce alkenyl, acetyl, or formyl, or together form a 5-7-membered heterocycle; excluding where Y7Gis N and R7Gis absent; Y8Gis C, R8Gis H, R2Gis NH2, R6Gis =0, and R1Gis H (unmodified guanine).

[0564] Unless otherwise indicated, modified thymine or modified uracil has structure (III):

[0565] III wherein: each X is independently O or S and R5Uis H, OH, halogen, 0-Ci-C2o alkyl, O-Ci-Ci2substituted alkyl, C1-C12 alkyl, substituted C1-C12 alkyl, C1-C12 alkenyl, substituted C1-C12 alkenyl, C1-C12 alkynyl, or substituted C1-C12 alkynyl; wherein if each X is O, R5Uis not H or CH, (unmodified uracil and unmodified thymine, respectively).

[0566] Unless otherwise indicated, modified cytosine has structure (IV): wherein: X is O or S; R4Cis N(Ra)(Rb); R5Cis H, OH, halogen, O-Ci-Ci2alkyl, O-Ci-Ci2substituted alkyl, Ci-Ci2alkyl , substituted C1-C12 alkyl, C1-C12 alkenyl, or substituted C1-C12 alkenyl; Raand Rbare CHEMO 113W0 independently H, Ci-Ce alkyl, substituted Ci-Ce alkyl, Ci-Ce alkenyl, substituted Ci-Ce alkenyl, C1-C12 alkynyl, substituted C1-C12 alkynyl, acetyl, or formyl, or together form a 5-7-membered heterocycle; excluding where X is O, R4Cis NH2, and R5Cis H (unmodified cytosine).

[0567] Hypoxanthine has structure (V):

[0568] V

[0569] Hypoxanthine is considered a modified adenine, where Y7Ais N and R7Ais absent; Y8Ais C, R8Ais H, R1Ais H, R2Ais H, and R6Ais oxo.

[0570] In certain embodiments, modified nucleobases of a modified oligonucleotide are selected from: 5- substituted pyrimidines, 6-azapyrimidines, alkyl or alkynyl substituted pyrimidines, alkyl substituted purines, and N-2, N-6, and 0-6 substituted purines. In certain embodiments, modified nucleobases are selected from: 5-methylcytosine, hypoxanthine, 1 -methylpseudouridine, 2-aminopropyladenine, 5- hydroxymethyl cytosine, xanthine, 2-aminoadenine, 6-N-methylguanine, 6-N-methyladenine, 2- propyladenine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-propynyl (-CYC -CH3) uracil, 5- propynylcytosine, 6-azouracil, 6-azocytosine, 6-azothymine, 5 -ribosyluracil (pseudouracil), 4-thiouracil, 8- halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl, 8-aza and other 8-substituted purines, 5-halo (particularly 5-bromo), 5 -trifluoromethyl, 5-halouracil, and 5-halocytosine, 7-methylguanine, 7-methyladenine, 2-F- adenine, 2-aminoadenine, 7-deazaguanine, 7-deazaadenine, 3-deazaguanine, 3 -deazaadenine, 6-N- benzoyladenine, 2-N-isobutyrylguanine, 4-N-benzoylcytosine, 4-N-benzoyluracil, 5-methyl 4-N- benzoylcytosine, 5-methyl 4-N-benzoyluracil, universal bases, hydrophobic bases, promiscuous bases, size-expanded bases, and fluorinated bases. Further modified nucleobases include tricyclic pyrimidines, such as l,3-diazaphenoxazine-2-one, l,3-diazaphenothiazine-2-one, and 9-(2 -aminoethoxy)- 1,3- diazaphenoxazine-2-one (G-clamp). Modified nucleobases may also include those in which the purine or pyrimidine base is replaced with other heterocycles, for example, 7-deaza-adenine, 7-deazaguanosine, 2- aminopyridine, and 2-pyridone. Further nucleobases include those disclosed in Englisch, U. et al., Angew. Chem. Int. Ed. 1991, 30, 613; Sanghvi, Y.S., Chapter 15, Antisense Research and Applications, Crooke, S. T. and Lebleu, B., Eds., CRC Press, 1993, 273-288; and those disclosed in Chapters 6 and 15, Antisense Drug Technology, Crooke S.T., Ed., CRC Press, 2008, 163-166 and 442-443.

[0571] Preparation of certain of the above noted modified nucleobases, as well as other modified nucleobases, are known in the art and can be readily identified in publications, including, without limitation, Rogers et al., U.S. 5,134,066 ; Benner et al., U.S. 5,432,272; Matteucci et al., U.S. 5,502,177 ; Froehler et al., M. . 5,594,121 ; and Cook et al., U.S. 5,681,941.

[0572] In certain embodiments, at least one nucleobase of a modified oligonucleotide is a modified nucleobase selected from modified adenine (A) having a structure represented by structure I, modified guanine (G) having a structure represented by structure II, modified thymine (T) or modified uracil (U) CHEMO 113W0 having a structure represented by structure III, and modified cytosine (C) having a structure represented by structure IV.

[0573] In certain embodiments, each nucleobase of a modified oligonucleotide is selected from unmodified A, unmodified G, unmodified C, unmodified T, unmodified U, and 5 -methylcytosine (mC). 5-methylcytosine is a modified nucleobase having structure IV, where X is O, R4Cis NH2, and R5Cis CH3.

[0574] In certain embodiments, each nucleobase of a modified oligonucleotide is selected from unmodified A, unmodified G, unmodified C, unmodified T, unmodified U, 5-methylcytosine (mC), and hypoxanthine. Hypoxanthine is a modified nucleobase having structure V and is also a modified A represented by structure I, where Y7Ais N and R7Ais absent; Y8Ais C, R8Ais H, R1Ais H, R2Ais H, and R6Ais oxo.

[0575] In certain embodiments, there are no modified nucleobases in a modified oligonucleotide and each nucleobase of a modified oligonucleotide is selected from unmodified A, unmodified G, unmodified C, unmodified T, and unmodified U.

[0576] 3. Modified Internucleoside Linkages

[0577] In certain embodiments, oligomeric agents provided herein comprise or consist of a modified oligonucleotide comprising at least one modified intemucleoside linkage. The naturally occurring intemucleoside linkage of RNA and DNA is a 3' to 5' phosphodiester linkage. Herein, all intemucleoside linkages between furanosyl sugar moieties are 3' to 5' intemucleoside linkages unless otherwise indicated. An intemucleoside linkage can link any two nucleosides, including unmodified nucleosides, modified nucleosides, and sugar surrogate nucleosides.

[0578] The two main classes of intemucleoside linkages are defined by the presence or absence of a phosphoms atom. Representative phosphoms-containing intemucleoside linkages include but are not limited to phosphates, which contain a phosphodiester bond (also referred to as unmodified linkages), phosphotriesters, methylphosphonates, phosphoramidates, phosphorothioates, and phosphorodithioates. Representative non-phosphorus containing intemucleoside linkages include but are not limited to methylenemethylimino (-CH2-N(CH3)-O-CH2-), thiodiester, thionocarbamate (-O-C(=O)(NH)-S-), siloxane (-O-S1H2-O-), and N,N'-dimethylhydrazine (-CH2-N(CH3)-N(CH3)-). Modified intemucleoside linkages, compared to naturally occurring phosphodiester linkages, may be used to alter, typically increase, nuclease resistance of the oligonucleotide.

[0579] In certain embodiments, a modified intemucleoside linkage is any of those described in WO 2021 / 030778. In certain embodiments, a modified intemucleoside linkage has the formula: wherein independently for each intemucleoside linkage of the modified oligonucleotide: X is selected from O and S;

[0580] Ri is selected from H, Ci-Ce alkyl, and substituted Ci-Ce alkyl; and CHEMO 113W0

[0581] T is selected from SO2R2, C(=O)R3, and P(=O)R4R5, wherein:

[0582] R2 is selected from an aryl, a substituted aryl, a heterocycle, a substituted heterocycle, an aromatic heterocycle, a substituted aromatic heterocycle, a diazole, a substituted diazole, a Ci-Ce alkoxy, Ci-Ce alkyl, Ci-Ce alkenyl, Ci-Ce alkynyl, substituted Ci-Ce alkyl, substituted Ci-Ce alkenyl, substituted Ci-Ce alkynyl, and a conjugate group;

[0583] Rs is selected from an aryl, a substituted aryl, CH3, N(CH3)2, OCH3, and a conjugate group;

[0584] R4 is selected from OCH3, OH, Ci-Ce alkyl, substituted Ci-Ce alkyl, and a conjugate group; and

[0585] Rs is selected from OCH3, OH, Ci-Ce alkyl, and substituted Ci-Ce alkyl.

[0586] In certain embodiments, a modified oligonucleotide comprises a mesyl phosphoramidate linkage having a formula:

[0587] Certain intemucleoside linkages having reduced charge (referred to as “neutral intemucleoside linkages”) have been described. Such neutral intemucleoside linkages include, without limitation, phosphotriesters, methylphosphonates, MMI (3'-CH2-N(CH3)-O-5'), amide-3 (3'-CH2-C(=O)-N(H)-5'), amide-4 (3'-CH2-N(H)-C(=O)-5'), formacetal (3'-O-CH2-O-5'), methoxypropyl (MOP) (see US 9,926,556), and thioformacetal (3'-S-CH2-O-5'). Further neutral intemucleoside linkages include nonionic linkages comprising siloxane (dialkylsiloxane), carboxylate ester, carboxamide, sulfide, sulfonate ester and amides (See for example: Carbohydrate Modifications in Antisense Research,' Y.S. Sanghvi and P.D. Cook, Eds., ACS Symposium Series 580; Chapters 3 and 4, 40-65). Further neutral intemucleoside linkages include nonionic linkages comprising mixed N, O, S and CH2 component parts.

[0588] In certain embodiments, a modified oligonucleotide comprises an intemucleoside linkage comprising a triazole, alkyne, or cyclic guanidine moiety. In certain embodiments, a modified oligonucleotide comprises an intemucleoside linkage having a formula: which may be stereorandom or may be enriched for the ( / p) or (.S'p) configuration.

[0589] In certain embodiments, intemucleoside linkages are not 3'-to-5' intemucleoside linkages.

[0590] In certain embodiments, modified oligonucleotides comprise one or more inverted nucleoside, where a sugar moiety is linked 3' to 3' and / or 5' to 5', as shown below: wherein each Bx independently represents any nucleobase.

[0591] In certain embodiments, an inverted nucleoside is terminal (z.e., the last nucleoside on one end of an oligonucleotide) and so only one intemucleoside linkage depicted above will be present. In certain embodiments, additional features (e.g., a conjugate group) are attached to the inverted nucleoside. Such terminal inverted nucleosides may be attached to either or both ends of an oligonucleotide.

[0592] In certain embodiments, inverted nucleosides lack a nucleobase (are abasic nucleosides). In certain such embodiments, additional features (e.g., a conjugate group) are attached to the inverted abasic nucleoside. A terminal inverted nucleoside may be attached to either or both ends of an oligonucleotide. In certain embodiments, nucleosides are linked 2' to 5' rather than the 3' to 5' linkage. Such a linkage is illustrated below. wherein each Bx represents any nucleobase. In certain embodiments, a bicyclic sugar moiety may be linked via an atom on the non-furanosyl ring. In certain such embodiments, a bicyclic sugar moiety is linked 7’ to 5’, as shown below:

[0593]

[0594] In certain embodiments, intemucleoside linkages have at least one chiral center. In such embodiments, a chiral atom can be prepared as a racemic mixture, or as separate enantiomers. Representative intemucleoside linkages having a chiral center include but are not limited to alkylphosphonates, mesyl phosphoramidates, and phosphorothioates.

[0595] The mesyl phosphoramidate intemucleoside linkage comprises a chiral center. In certain embodiments, modified oligonucleotides comprising (Rp) and / or (.S'p) mesyl phosphoramidates comprise one or more of the following formulas, respectively, wherein “Bx” indicates a nucleobase:

[0596] The phosphorothioate intemucleoside linkage comprises a chiral center. In certain embodiments, modified oligonucleotides comprising (Rp) and / or (.S'p) phosphorothioates comprise one or more of the following formulas, respectively, wherein “Bx” indicates a nucleobase: CHEMO 113W0

[0597] Modified oligonucleotides comprising intemucleoside linkages having a chiral center may be prepared as populations of modified oligonucleotides comprising stereorandom intemucleoside linkages, or as populations of modified oligonucleotides comprising intemucleoside linkages containing chiral centers in particular stereochemical configurations. In certain embodiments, populations of modified oligonucleotides comprise one or more phosphorothioate intemucleoside linkages wherein all of the phosphorothioate intemucleoside linkages are stereorandom. In certain embodiments, populations of modified oligonucleotides comprise one or more mesyl phosphoramidate intemucleoside linkages wherein all of the mesyl phosphoramidate intemucleoside linkages are stereorandom. Such modified oligonucleotides can be generated using synthetic methods that result in random selection of the stereochemical configuration of each phosphorothioate and / or mesyl phosphoramidate linkage. Nonetheless, each individual phosphorothioate and / or mesyl phosphoramidate of each individual oligonucleotide molecule has a defined stereoconfiguration. In certain embodiments, populations of modified oligonucleotides are enriched for modified oligonucleotides comprising one or more particular phosphorothioate and / or mesyl phosphoramidate intemucleoside linkages in a particular, independently selected stereochemical configuration (e.g., Rp or .S'p), In certain embodiments, the particular phosphorothioate and / or mesyl phosphoramidate linkage is present in the selected configuration in at least 65%, 70%, 80, 90%, or 99% of the molecules in the population. Such chirally enriched populations of modified oligonucleotides can be generated using synthetic methods known in the art, e.g., methods described in Oka, N., etal. J. Am. Chem. Soc. 2003, 125, 8307-8317; Wan, W. B., et al. Nucleic Acids Res . 2014, 42, 13456. and WO 2017 / 015555.

[0598] As used herein, “chirally enriched” in reference to a population means a plurality of molecules of identical molecular formula, wherein one or more particular chiral centers are not stereorandom as defined herein. Chirally enriched stereocenters are intentionally controlled during synthesis, or enriched following synthesis, for a particular absolute stereochemical configuration at that center. Populations of molecules having multiple chiral centers within each molecule may contain one or more stereorandom chiral centers. In certain embodiments, the molecules are modified oligonucleotides. In certain embodiments, the chiral center is at the phosphorous atom of a phosphorothioate intemucleoside linkage. In certain embodiments, the chiral center is at the phosphorous atom of a mesyl phosphoramidate intemucleoside linkage. In certain embodiments, a population of modified oligonucleotides is enriched for modified oligonucleotides having CHEMO 113W0 at least one indicated phosphorothioate and / or mesyl phosphoramidate in the (.S'p) configuration. In certain embodiments, a population of modified oligonucleotides is enriched for modified oligonucleotides having at least one phosphorothioate and / or mesyl phosphoramidate in the (Rp) configuration. Unless otherwise indicated, chiral intemucleoside linkages of modified oligonucleotides described herein may be stereorandom or chirally enriched.

[0599] B. Motifs

[0600] A modified oligonucleotide may be described by its sugar motif, nucleobase motif, and / or intemucleoside linkage motif (as used herein, nucleobase motif describes the modifications to the nucleobases independent of the nucleobase sequence). For a given oligonucleotide, the, sugar motif, nucleobase motif, and intemucleoside linkage motif are conceptually separable from each other, and with nucleobase sequence, and thus each may be described independently.

[0601] 1. Sugar Motifs

[0602] In certain embodiments, oligonucleotides comprise one or more type of modified sugar moiety and / or unmodified sugar moiety arranged along the oligonucleotide or region thereof in a defined pattern or sugar motif. In certain instances, such sugar motifs include but are not limited to any of the modified sugar moieties discussed herein. In certain embodiments, the sugar moiety of at least one nucleoside of a sense oligonucleotide is a modified sugar moiety.

[0603] In certain embodiments, a modified oligonucleotide comprises a deoxy region. In certain embodiments, each nucleoside of the deoxy region is a deoxynucleoside. In certain embodiments, each nucleoside of the deoxy region is a 2'-P-D-deoxynucleoside. In certain embodiments, the deoxy (G) region consists of 5-20 linked nucleosides. In certain embodiments, the deoxy (G) region consists of 5, 6, 7, 8, 9, 10, 7-15, or 11-20 linked nucleosides. In certain embodiments, the deoxy (G) region consists of 10-13 linked nucleosides, optionally 10, optionally 11, optionally 12, or optionally 13 linked nucleosides. In certain embodiments, the deoxy (G) region consists of 11-15 linked nucleosides, optionally 11, optionally 12, optionally 13, optionally 14, or optionally 15 linked nucleosides. In certain embodiments, the deoxy (G) region consists of 10 linked nucleosides.

[0604] In certain embodiments, a modified oligonucleotide comprises or consists of a region of formula I (5’ to 3’):

[0605] Ab-Wl-G-W2

[0606] I wherein Ab is a bicyclic adenosine and G is a deoxy region. In certain embodiments, Ab is a cEt adenosine. In certain embodiments, the deoxy region is flanked on the 5 '-side by a region W1 consisting of linked nucleosides and on the 3 '-side by a region W2 consisting of linked nucleosides, wherein the 3 '-most nucleoside of the W1 region comprises a modified sugar moiety and the 5 '-most nucleoside of the W2 region comprises a modified sugar moiety. The three regions (Wl, G, W2) may form a contiguous sequence of nucleosides. In certain embodiments, each nucleoside of the Wl region or each nucleoside of the W2 region comprises a modified sugar moiety. In certain embodiments, at least two nucleosides of the Wl CHEMO 113W0 region and at least two nucleosides of the W2 region comprises a modified sugar moiety. In certain embodiments, at least three nucleosides of the W1 region and at least three nucleosides of the W2 region comprises a modified sugar moiety. In certain embodiments, at least four nucleosides of the W1 region comprises a modified sugar moiety. In certain embodiments, each of the nucleosides within the W 1 region comprise the same modified sugar moiety. In certain embodiments, nucleosides within the W1 region comprise two or more different modified sugar moieties. In certain embodiments, each of the nucleosides within the W2 region comprise the same modified sugar moiety. In certain embodiments, the nucleosides within the W2 region comprise two or more different modified sugar moieties.

[0607] In certain embodiments, the W1 region and the W2 region of a modified oligonucleotide each consist of 1-8 nucleosides. In certain embodiments, the W1 region consists of 1-7 nucleosides. In certain embodiments, the W1 region consists of 1-6, 2-6, 3-6, or 3-5 nucleosides. In certain embodiments, the W1 region consists of 1, 2, 3, 4, 5, 6, 7, or 8 nucleosides. In certain embodiments, the W2 region consists of 1- 7 nucleosides. In certain embodiments, the W2 region consists of 1-6, 2-6, 3-6, or 3-5 nucleosides. In certain embodiments, the W2 region consists of 1, 2, 3, 4, 5, 6, 7, or 8 nucleosides.

[0608] In certain embodiments, such modified oligonucleotides are referred to as “gapmers”. Herein, the lengths (number of nucleosides) of the W1 region, the deoxy region, and the W2 region of an oligonucleotide may be provided using the notation [# of nucleosides in the W 1 region] - [# of nucleosides in the deoxy region] - [# of nucleosides in the W2 region]. Thus, a 3-10-3 gapmer consists of 3 linked nucleosides in each wing and 10 linked nucleosides in the gap (not including a linked 5 ’-bicyclic adenosine). Where such nomenclature is followed by a specific modification, that modification is the modification in each sugar moiety of each nucleoside of the W 1 region and the W2 region and the gap nucleosides comprise 2'-P-D-deoxyribosyl sugar moieties. Thus, a 5-10-5 MOE gapmer consists of 5 linked 2'-M0E nucleosides in the W1 region (or “5’-wing”), 10 linked 2'-P-D-deoxynucleosides in the deoxy region (or “gap”), and 5 linked 2'-M0E nucleosides in the W2 region (or “3 ’-wing”) (not including the linked 5 ’-cEt adenosine). A “mixed” gapmer has at least two differently modified sugar moieties in the 5'- and / or the 3 '-regions.

[0609] In certain embodiments, the region of formula I is contiguous with a second nucleoside region that, at least in part, forms a duplex with some or all of the nucleosides of the region of formula I. In certain embodiments, the second nucleoside region is 10-25 nucleosides in length.

[0610] In certain embodiments, the region of formula I is contiguous with linker-nucleosides as described herein and known in the art. The linker-nucleosides may form a cleavable moiety that links the modified oligonucleotide to a conjugate moiety or conjugate group.

[0611] In certain embodiments, provided is an oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0612] Ab-Wl-G-W2

[0613] I CHEMO 113W0 wherein Ab is a bicyclic adenosine, W1 consists of 1-7 linked nucleosides, wherein each nucleoside of W1 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxynucleoside, provided that the 3 ’-most nucleoside of W1 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;

[0614] G consists of 5-20 linked 2’-deoxynucleosides;

[0615] W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; optionally wherein the modified oligonucleotide consists of the region of formula I.

[0616] In certain embodiments, each intemucleoside linkage within G is independently selected from a mesyl phosphoramidate and a phosphorothioate. In certain embodiments, W1 does not comprise a sugar surrogate nucleoside. In certain embodiments, W1 comprises one sugar surrogate nucleoside. In certain embodiments, W2 does not comprise a sugar surrogate nucleoside. In certain embodiments, W2 comprises one sugar surrogate nucleoside. In certain embodiments, at least one of W1 and W2 comprises a 2’- deoxynucleoside. In certain embodiments, W1 comprises a 2 ’-deoxynucleoside. In certain embodiments, W2 comprises a 2 ’-deoxynucleoside. In certain embodiments, G consists of at least 11, optionally 11-20, optionally 11-15, contiguous nucleosides. In certain embodiments, the oligomeric agent comprises the modified oligonucleotide, a conjugate group, and optionally a conjugate linker. In certain embodiments, the oligomeric agent consists of the modified oligonucleotide. In certain embodiments, the modified oligonucleotide is a first modified oligonucleotide and the oligomeric agent comprises a second oligonucleotide that is complimentary to and at least partially duplexed with the first modified oligonucleotide.

[0617] In certain embodiments, each nucleoside of W1 is independently selected from a bicyclic nucleoside other than a cEt nucleoside, a 2 ’-substituted nucleoside, a sugar surrogate nucleoside, and a 2’- deoxynucleoside.

[0618] In certain embodiments, at least one nucleoside of W1 is not a cEt.

[0619] In certain embodiments, the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to an Lpa transcript (SEQ ID NO: 1).

[0620] In certain embodiments, the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to an APOE transcript (SEQ ID NO: 2).

[0621] In certain embodiments, the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to a PLN transcript (SEQ ID NO: 3). In certain embodiments, the 3 ’-most nucleoside of W1 does not comprise a 2’-OMe nucleoside. CHEMO 113W0

[0622] In certain embodiments, the modified oligonucleotide comprises at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate.

[0623] In certain embodiments, W1 does not comprise a cEt nucleoside.

[0624] In certain embodiments, the modified oligonucleotide is not compound 566830:

[0625] AksGdsGksCdsAksTdsAdsGdsCdsAdsGdsCdsAdsGdsGesAe (SEQ ID NO: 4) wherein

[0626] A is an adenine nucleobase,

[0627] C is a 5 -methylcytosine nucleobase,

[0628] G is a guanine nucleobase,

[0629] T is a thymine nucleobase, k is a cEt sugar moiety, d is a 2 ’-deoxyribosyl sugar moiety, e is a 2 ’-MOE sugar moiety, and s is a phosphorothioate intemucleoside linkage.

[0630] In certain embodiments, the modified oligonucleotide comprises at least one phosphorothioate intemucleoside linkage.

[0631] In certain embodiments, the modified oligonucleotide at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate.

[0632] In certain embodiments, the modified oligonucleotide comprises no more than three 2’-M0E nucleosides.

[0633] In certain embodiments, the modified oligonucleotide comprises no more than three cEt nucleosides.

[0634] In certain embodiments, W2 comprises at least two non-cEt nucleosides. In certain embodiments, W2 comprises two, three, four, or five 2’-MOE nucleosides. In certain embodiments, W2 comprises four contiguous 2 ’-MOE nucleosides. In certain embodiments, W2 consists of five contiguous 2 ’-MOE nucleosides.

[0635] In certain embodiments, the modified oligonucleotide comprises at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate.

[0636] In certain embodiments, W1 or W2 comprises at least one 2-deoxynucleoside.

[0637] In certain embodiments, the modified oligonucleotide comprises no more than three cEt nucleosides.

[0638] In certain embodiments, the modified oligonucleotide comprises no more than three 2’-MOE nucleosides.

[0639] In certain embodiments, the modified oligonucleotide comprises at least four 2’-MOE nucleosides.

[0640] In certain embodiments, W1 comprises no more than three 2’-MOE nucleosides.

[0641] In certain embodiments, W1 does not comprise three contiguous cEt nucleosides.

[0642] In certain embodiments, W1 does not comprise three contiguous 2’-MOE nucleosides. CHEMO 113W0

[0643] In certain embodiments, W1 comprises at least four 2’-M0E nucleosides. In certain embodiments, each nucleoside of W1 is a 2 ’-MOE nucleoside. In certain embodiments, W1 consists of four contiguous 2’-M0E nucleosides.

[0644] In certain embodiments, provided is an oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0645] Ab-Wl-G-W2

[0646] I wherein Ab is a bicyclic adenosine, W1 consists of 1-7 nucleosides, each independently selected from among a bicyclic nucleoside and a 2'-P-D-deoxyribosyl nucleoside, wherein the 3 ’-most nucleoside of W1 is a bicyclic nucleoside; wherein G consists of 5-20 2’-P-D-deoxynucleosides linked by mesyl phosphoramidate and / or phosphorothioate intemucleoside linkages; wherein W2 consists of 2-8 nucleosides, each independently selected from among a bicyclic nucleoside and a 2'-p-D-deoxyribosyl nucleoside, wherein the 5’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; provided that at least one of W1 and W2 comprises a 2’-P-D-deoxyribosyl nucleoside, and each 2’-P-D-deoxyribosyl nucleoside ofWl and W2 is linked at its 3 ’-position by a mesyl phosphoramidate intemucleoside linkage. In certain embodiments, each bicyclic nucleoside is selected from cEt and LNA.

[0647] In certain embodiments,

[0648] W2 has the formula -(E)n4-(D4)n5-(F)n6 wherein each F is independently selected from a bicyclic nucleoside, a 2 ’-substituted nucleoside, and a 2 ’-deoxy nucleoside, provided that at least one F is a bicyclic nucleoside or a 2 ’-substituted nucleoside; and each non-terminal 2’-deoxy nucleoside of F is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; each E is independently a bicyclic nucleoside or a 2 ’-substituted nucleoside; each D4 is independently a 2 ’-deoxy nucleoside, provided that each D4 is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; n6 is 1 to 8 and each of n4 and n5 is independently 0 to 8.

[0649] In certain embodiments,

[0650] W1 has the formula (A)nl-(Dl)n2-(B)n3- wherein each A is independently selected from a bicyclic nucleoside, a 2 ’-substituted nucleoside, and a 2 ’-deoxy nucleoside provided that at least one A is a bicyclic nucleoside or a 2 ’-substituted nucleoside; and each 2’-deoxy nucleoside of A is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; each B is independently a bicyclic nucleoside or a 2 ’-substituted nucleoside; each DI is independently a 2 ’-deoxy nucleoside provided that each DI is linked at its 3 ’-position by a CHEMO 113W0 phosphorothioate or mesyl phosphoramidate intemucleoside linkage; nl is 1 to 8 and each of n2 and n3 is independently 0 to 8.

[0651] In certain embodiments,

[0652] G consists of 5-20 linked 2’-deoxynucleosides; provided that each nucleoside of G is linked at its 3 ’-position by a mesyl phosphoramidate or a phosphorothioate intemucleoside linkage.

[0653] In certain embodiments, the sum of n4, n5, and n6 is no more than 8. In certain embodiments, the sum of nl, n2, and n3 is no more than 8.

[0654] In certain embodiments, provided is an oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0655] Ab-Wl-G-W2

[0656] I wherein Ab is a bicyclic adenosine, W1 consists of 3, 4, or 5 nucleosides, each independently selected from among a bicyclic nucleoside and a 2'-P-D-deoxyribosyl nucleoside, wherein the 3 ’-most nucleoside of W1 is a bicyclic nucleoside; wherein G consists of 7-15 2’-p-D-deoxynucleosides linked by mesyl phosphoramidate and / or phosphorothioate intemucleoside linkages; wherein W2 consists of 3, 4, 5, or 6 nucleosides, each independently selected from among a bicyclic nucleoside and a 2'-p-D- deoxyribosyl nucleoside, wherein the 5 ’-most nucleoside of W2 is a bicyclic nucleoside; provided that at least one ofWl and W2 comprises a 2’-P-D-deoxyribosyl nucleoside, and each 2’-P-D-deoxyribosyl nucleoside of W1 and W2 is linked at its 3 ’-position by a mesyl phosphoramidate intemucleoside linkage. In certain embodiments, each bicyclic nucleoside is selected from cEt and LNA.

[0657] In certain embodiments, provided is an oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):

[0658] Ab-Wl-G-W2

[0659] I wherein Ab is a bicyclic adenosine, W 1 consists of 4 linked nucleosides, wherein each nucleoside of W1 is a 2 ’-substituted nucleoside;

[0660] G consists of 10 linked 2’-deoxynucleosides;

[0661] W2 consists of 5 linked nucleosides, wherein each nucleoside of W2 is independently selected from a 2 ’-substituted nucleoside and a bicyclic nucleoside; optionally wherein the modified oligonucleotide consists of the region of formula I. CHEMO 113W0

[0662] In certain embodiments, each 2 ’-substituted nucleoside is selected from a 2 ’-MOE nucleoside and a 2’-OMe nucleoside. In certain embodiments, each 2 ’-substituted nucleoside is a 2 ’-MOE nucleoside.

[0663] In certain embodiments, each bicyclic nucleoside is selected from a cEt nucleoside and an LNA nucleoside. In certain embodiments, each bicyclic nucleoside is a cEt nucleoside.

[0664] In certain embodiments, W2 comprises no bicyclic nucleosides. In certain embodiments, W2 comprises exactly one bicyclic nucleoside. In certain embodiments, W2 comprises exactly two bicyclic nucleosides.

[0665] In certain embodiments, provided is an oligomeric agent comprising a modified oligonucleotide consisting of 14 to 20 linked nucleosides, wherein the modified oligonucleotide has formula I (5’ to 3’):

[0666] Ab-Wl-G-W2

[0667] I wherein Ab is a bicyclic adenosine, W1 has the formula (A)nl-(Dl)n2-(B)n3- wherein each A is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’ -deoxy nucleoside provided that at least one A is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; and each 2’- deoxy nucleoside of A is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; each B is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety and a sugar surrogate nucleoside; each DI is independently a 2 ’-deoxy nucleoside provided that each DI is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; nl is 1 to 7 and each of n2 and n3 is independently 0 to 7;

[0668] G consists of 5-20 linked 2’-deoxynucleosides; provided that each nucleoside of G is linked at its 3’- position by a mesyl phosphoramidate or a phosphorothioate intemucleoside linkage; wherein W2 has the formula -(E)n4-(D4)n5-(F)n6 wherein each F is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxy nucleoside, provided that at least one F is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; and each non-terminal 2 ’-deoxy nucleoside of F is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; each E is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety and a sugar surrogate nucleoside; each D4 is independently a 2 ’-deoxy nucleoside provided that each D4 is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; CHEMO 113W0 n6 is 1 to 8 and each of n4 and n5 is independently 0 to 8.

[0669] In certain embodiments, 1, 2, 3, 4, 5, 6, 7, or 8 nucleosides of G are linked by 3’ mesyl phosphoramidate intemucleoside linkages. In certain embodiments, 1, 2, 3, 4, 5, 6, 7, or 8 contiguous nucleosides of G are linked by 3’ mesyl phosphoramidate intemucleoside linkages. In certain embodiments, 2-6 nucleosides of G are linked by 3’ mesyl phosphoramidate intemucleoside linkages. In certain embodiments, 3-5 nucleosides of G are linked by 3’ mesyl phosphoramidate intemucleoside linkages. In certain embodiments, 2-6 contiguous nucleosides, optionally 2-4 contiguous nucleosides, of G are linked by 3’ mesyl phosphoramidate intemucleoside linkages.

[0670] 2. Intemucleoside Linkage Motifs

[0671] In certain embodiments, oligonucleotides comprise modified and unmodified intemucleoside linkages arranged along the oligonucleotide or region thereof in a defined pattern or motif. In certain embodiments, each intemucleoside linkage of a modified oligonucleotide is independently selected from a phosphorothioate intemucleoside linkage, a mesyl phosphoramidate intemucleoside linkage, and a phosphodiester intemucleoside linkage. In certain embodiments, each intemucleoside linkage of a modified oligonucleotide is independently selected from a mesyl phosphoramidate intemucleoside linkage and a phosphorothioate intemucleoside linkage. In certain embodiments, each phosphorothioate intemucleoside linkage is independently selected from a stereorandom phosphorothioate, a (.S'p) phosphorothioate, and a ( / ?p) phosphorothioate. In certain embodiments, each mesyl phosphoramidate intemucleoside linkage is independently selected from a stereorandom mesyl phosphoramidate, a (.S'p) mesyl phosphoramidate, and a ( / ?p) mesyl phosphoramidate.

[0672] In certain embodiments, the modified oligonucleotide has an intemucleoside linkage motif as exemplified in one or more compounds described herein. In certain embodiments, each 2 ’-deoxynucleoside of W1 is linked at its 3 ’-position by a mesyl phosphoramidate intemucleoside linkage. In certain embodiments, each 2 ’-deoxynucleoside of W2 is linked at its 3’-position by a mesyl phosphoramidate intemucleoside linkage. In certain embodiments, 1, 2, 3, 4, 5, 6, 7, or 8 2 ’-deoxynucleoside of G is linked at its 3 ’-position by a mesyl phosphoramidate intemucleoside linkage. In certain embodiments, the modified oligonucleotide comprises only mesyl phosphoramidate, phosphodiester, and phosphorothioate intemucleoside linkages. In certain embodiments, the modified oligonucleotide comprises 1, 2, 3, 4, 5, or 6 phosphodiester intemucleoside linkages. In certain embodiments, the modified oligonucleotide comprises 10-15 phosphorothioate and 5-10 mesyl phosphoramidate intemucleoside linkages. In certain embodiments, the modified oligonucleotide comprises 10-15 phosphorothioate and 5-10 mesyl phosphoramidate intemucleoside linkages. In certain embodiments, the modified oligonucleotide comprises only mesyl phosphoramidate and phosphorothioate intemucleoside linkages. In certain CHEMO 113W0 embodiments, the modified oligonucleotide comprises 11, 12, or 13 phosphorothioate and 6, 7, or 8 mesyl phosphoramidate intemucleoside linkages.

[0673] C. Lengths

[0674] It is possible to increase or decrease the length of an oligonucleotide without eliminating activity. For example, in Woolf et al. Proc. Natl. Acad. Set. USA 1992, 89:7305-7309, a series of oligonucleotides 13-25 nucleobases in length were tested for their ability to induce cleavage of a target RNA in an oocyte injection model. Oligonucleotides 25 nucleobases in length with 8 or 11 mismatch bases near the ends of the oligonucleotides were able to direct specific cleavage of the target RNA, albeit to a lesser extent than the oligonucleotides that contained no mismatches. Similarly, target specific cleavage was achieved using 13 nucleobase oligonucleotides, including those with 1 or 3 mismatches.

[0675] In certain embodiments, oligonucleotides (including modified oligonucleotides) have any of a variety of ranges of lengths. In certain embodiments, oligonucleotides consist of X to Y linked nucleosides, where X represents the fewest number of nucleosides in the range and Y represents the largest number nucleosides in the range. In certain such embodiments, X and Y are each independently selected from 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, and 50; provided that X< Y. For example, in certain embodiments, oligonucleotides consist of 14 to 15, 14 to 16, 14 to 17, 14 to 18, 14 to 19, 14 to 20, 14 to 21, 14 to 22, 14 to 23, 14 to 24, 14 to 25, 14 to 26, 14 to 27, 14 to 28, 14 to 29, 14 to 30, 15 to 16, 15 to 17, 15 to 18, 15 to 19, 15 to 20, 15 to 21, 15 to 22, 15 to 23, 15 to 24, 15 to 25, 15 to 26, 15 to 27, 15 to 28, 15 to 29, 15 to 30, 16 to 17, 16 to 18, 16 to 19, 16 to 20, 16 to 21, 16 to 22, 16 to 23, 16 to 24, 16 to 25, 16 to 26, 16 to 27, 16 to 28, 16 to 29, 16 to 30, 17 to 18, 17 to 19, 17 to 20, 17 to 21, 17 to 22, 17 to 23, 17 to 24, 17 to 25, 17 to

[0676] 26, 17 to 27, 17 to 28, 17 to 29, 17 to 30, 18 to 19, 18 to 20, 18 to 21, 18 to 22, 18 to 23, 18 to 24, 18 to 25,

[0677] 18 to 26, 18 to 27, 18 to 28, 18 to 29, 18 to 30, 19 to 20, 19 to 21, 19 to 22, 19 to 23, 19 to 24, 19 to 25, 19 to 26, 19 to 27, 19 to 28, 19 to 29, 19 to 30, 20 to 21, 20 to 22, 20 to 23, 20 to 24, 20 to 25, 20 to 26, 20 to

[0678] 27, 20 to 28, 20 to 29, 20 to 30, 21 to 22, 21 to 23, 21 to 24, 21 to 25, 21 to 26, 21 to 27, 21 to 28, 21 to 29,

[0679] 21 to 30, 22 to 23, 22 to 24, 22 to 25, 22 to 26, 22 to 27, 22 to 28, 22 to 29, 22 to 30, 23 to 24, 23 to 25, 23 to 26, 23 to 27, 23 to 28, 23 to 29, 23 to 30, 24 to 25, 24 to 26, 24 to 27, 24 to 28, 24 to 29, 24 to 30, 25 to

[0680] 26, 25 to 27, 25 to 28, 25 to 29, 25 to 30, 26 to 27, 26 to 28, 26 to 29, 26 to 30, 27 to 28, 27 to 29, 27 to 30, 28 to 29, 28 to 30, or 29 to 30 linked nucleosides.

[0681] In certain embodiments, modified oligonucleotides consist of 16 linked nucleosides. In certain embodiments, modified oligonucleotides consist of 17 linked nucleosides. In certain embodiments, modified oligonucleotides consist of 18 linked nucleosides. In certain embodiments, modified oligonucleotides consist of 19 linked nucleosides. In certain embodiments, modified oligonucleotides consist of 20 linked nucleosides. In certain embodiments, modified oligonucleotides consist of 21 linked nucleosides. In certain embodiments, modified oligonucleotides consist of 22 linked nucleosides. In certain embodiments, modified oligonucleotides consist of 23 linked nucleosides. In certain embodiments, the CHEMO 113W0 modified oligonucleotides have no more than 3, optionally no more than 1, mismatches to a target nucleic acid.

[0682] In certain embodiments, an oligonucleotide consists of 14-30 linked nucleosides. In certain embodiments, an oligonucleotide consists of 17-25, 17-23, 17-21, 18-30, 18-25, or 21-23 linked nucleosides. In certain embodiments, an oligonucleotide consists of 20 linked nucleosides.

[0683] D. Nucleobase Sequence

[0684] In certain embodiments, oligonucleotides (unmodified or modified oligonucleotides) are further described by their nucleobase sequence. In certain embodiments, oligonucleotides have a nucleobase sequence that is complementary to a nucleobase sequence of a second strand of linked nucleosides (e.g., another oligonucleotide or an identified reference nucleic acid, such as a target nucleic acid) or a region thereof. In certain embodiments, a region of an oligonucleotide has a nucleobase sequence that is complementary to a nucleobase sequence of a second strand of linked nucleosides or a region thereof. In certain embodiments, the nucleobase sequence of a region or entire length of an oligonucleotide is at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% complementary to the nucleobase sequence of a second strand of linked nucleosides or region thereof.

[0685] E. Oligomeric Duplexes

[0686] In certain embodiments, an oligomeric agent provided herein comprises a modified oligonucleotide paired (duplexed) with a second oligonucleotide to form an oligomeric duplex. In some embodiments, an oligomeric duplex comprises a first modified oligonucleotide having a targeting region complementary to a target nucleic acid and a second oligonucleotide having a duplexing region complementary to the first modified oligonucleotide or a region thereof. In certain embodiments, the second oligonucleotide is a modified oligonucleotide as described herein.

[0687] In certain embodiments, an oligomeric duplex comprises: a first modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the nucleobase sequence of the first modified oligonucleotide comprises at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, or at least 23 contiguous nucleosides; and a second modified oligonucleotide consisting of 6 to 30 linked nucleosides. In certain embodiments, the nucleobase sequence of the second modified oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, or 100% complementary to the nucleobase sequence of an equal length portion of the first modified oligonucleotide.

[0688] In certain embodiments, in an oligomeric agent comprising an oligomeric duplex provided herein, the first modified oligonucleotide of the oligomeric duplex is attached to a conjugate group. In certain embodiments, the conjugate group is attached to the first modified oligonucleotide at the 5 '-terminus of the first modified oligonucleotide. In certain embodiments, the conjugate group is attached to the first modified oligonucleotide at the 3 '-terminus of the first modified oligonucleotide. In certain embodiments, the conjugate group is attached to the first modified oligonucleotide at an internal position. In certain embodiments, the conjugate group is attached to the first modified oligonucleotide through a 2'- CHEMO 113W0 modification of a furanosyl sugar moiety. In certain embodiments, the conjugate group is attached to the first modified oligonucleotide through a modified intemucleoside linkage.

[0689] In certain embodiments, in an oligomeric agent comprising an oligomeric duplex provided herein, the second modified oligonucleotide of the oligomeric duplex is attached to a conjugate group. In certain embodiments, the conjugate group is attached to the second modified oligonucleotide at the 5'- terminus of the second modified oligonucleotide. In certain embodiments, the conjugate group is attached to the second modified oligonucleotide at the 3'- terminus of the second modified oligonucleotide. In certain embodiments, the conjugate group is attached to the second modified oligonucleotide at an internal position. In certain embodiments, the conjugate group is attached to the second modified oligonucleotide through a 2'-modification of a furanosyl sugar moiety. In certain embodiments, the conjugate group is attached to the second modified oligonucleotide through a modified intemucleoside linkage.

[0690] In certain embodiments, an oligomeric agent comprises two or more targeting regions. In certain embodiments, an oligomeric agent comprises two targeting regions.

[0691] In certain embodiments, an oligomeric agent comprises at least two oligomeric duplexes linked together. In certain embodiments, an oligomeric agent comprises two or more of the same oligomeric duplex, which is any of the oligomeric duplexes described herein. In certain embodiments, the two or more oligomeric duplexes are covalently linked together. In certain embodiments, the second modified oligonucleotides of the two or more oligomeric duplexes are covalently linked together. In certain embodiments, the second modified oligonucleotides of two or more oligomeric duplexes are covalently linked together at their 3' ends. In certain embodiments, the second modified oligonucleotides of two or more oligomeric duplexes are covalently linked together at the 3' end of one to the 5' end of the other. In certain embodiments, the two or more oligomeric duplexes are covalently linked together by a glycol linker, such as a tetraethylene glycol linker. A structure of oligomeric duplexes covalently linked by a glycol linker is described in, e.g., Alterman, J. F., et al. Nature Biotech. 2019, 37, 844-894. In certain embodiments, an oligomeric agent comprises two oligomeric duplexes formed from one antisense oligonucleotide comprising two targeting regions, wherein each targeting region is paired with a sense oligonucleotide. In some embodiments, an oligomeric agent comprises two or more oligomeric duplexes linked, e.g., covalently linked, together in a branched structure, e.g., a di-branched, tri-branched, or tetra-branched structure (see, e.g., WO 2022 / 256565). In some such embodiments, the structure contains a linker (e.g., one or more subunits of an ethylene glycol, alkyl, carbohydrate, block copolymer, peptide, ester, amide, carbamate, triazole) and optionally one or more branch point moieties (e.g., phosphoramidite, tosylated solketal, 1,3-diaminopropanol, pentaerythritol).

[0692] In some embodiments, an oligomeric duplex comprises a first modified oligonucleotide comprising a nucleobase sequence complementary to a first nucleobase sequence in a nucleic acid (the “targeting region”) and a second modified oligonucleotide comprising a nucleobase sequence complementary to: (a) a different (i. e. , second) sequence in a nucleic acid, or (b) a sequence in a target nucleic acid other than a nucleic acid. In some such embodiments, the first modified oligonucleotide further comprises a complementary region having a nucleobase sequence complementary to an equal length portion of the CHEMO 113W0 nucleobase sequence in the second modified oligonucleotide that is complementary to the second sequence in a nucleic acid (or to an equal length portion of the nucleobase sequence that is complementary to a sequence in a target nucleic acid other than a nucleic acid) . In some such embodiments, the second modified oligonucleotide further comprises a nucleobase sequence complementary to an equal length portion of the nucleobase sequence in the first modified oligonucleotide that is complementary to the first sequence in a nucleic acid (see, e.g., WO 2020 / 065602).

[0693] G. Conjugates

[0694] In certain embodiments, provided herein are oligomeric agents comprising one or more modified oligonucleotides, and one or more conjugate groups and / or one or more terminal groups. A conjugate group may be attached at the 5' terminus of an oligonucleotide and / or at the 3' terminus of an oligonucleotide and / or at any internal position of an oligonucleotide. In certain embodiments, conjugate groups are attached through a modified sugar moiety or a modified intemucleoside linkage. In certain embodiments, the conjugate moiety is a cell-targeting moiety.

[0695] 1. Conjugate Groups

[0696] A conjugate group comprises or consists of a conjugate moiety and a conjugate linker. A conjugate moiety attached to an oligonucleotide modifies one or more properties of the attached oligonucleotide compared to the same oligonucleotide lacking the conjugate moiety, including but not limited to pharmacodynamics, pharmacokinetics, stability, binding, absorption, tissue distribution, cellular distribution, cellular uptake, charge, and clearance. In certain embodiments, a conjugate moiety imparts a new property on the attached oligonucleotide.

[0697] In some embodiments, the conjugate group comprises a small molecule drug (e.g., an active pharmaceutical ingredient), an aliphatic chain, a lipid, a peptide, a protein, a hydrocarbon, a polyamine, a polyamide, a polyether, a thioether, an aptamer, an antibody, an antibody fragment, a VHH camelid antibody fragment, a VNAR shark antibody fragment, a vitamin, a fatty acid, a carbohydrate, an intercalator, a reporter molecule, a small molecule, or an alkyl moiety, e.g., a C22 alkyl, C20 alkyl, C17 alkyl, C16 alkyl, CIO alkyl, C21 alkyl, C19 alkyl, C18 alkyl, C15 alkyl, C14 alkyl, C13 alkyl, C12 alkyl, Cl 1 alkyl, C9 alkyl, C8 alkyl, C7 alkyl, C6 alkyl, or C5 alkyl, wherein the alkyl chain optionally has one or more unsaturated bonds. In some embodiments, the conjugate group comprises a 6-palmitamidohexyl moiety or a 2-(hydroxymethyl)-6-palmitamidohexyl moiety. In certain embodiments, the conjugate group comprises a cell-targeting moiety.

[0698] In certain embodiment, the conjugate group comprises a lipophilic moiety. In certain embodiments, the lipophilic moiety is selected from the group consisting of a C8-C20 alkyl moiety, cholesterol, retinoic acid, cholic acid, adamantane acetic acid, 1 -pyrene butyric acid, dihydrotestosterone, 1,3-bis- O(hexadecyl)glycerol, geranyloxyhexyanol, hexadecylglycerol, borneol, menthol, 1,3 -propanediol, heptadecyl group, palmitic acid, myristic acid, O3-(oleoyl) lithocholic acid, O3-(oleoyl)cholenic acid, dimethoxytrityl, or phenoxazine. In certain embodiments, the alkyl moiety is a saturated straight chain C16 hydrocarbon. CHEMO 113W0

[0699] Methods of preparing conjugated oligonucleotides are known in the art and / or described herein. For example, in one non-limiting solid phase method for large-scale synthesis of conjugated oligonucleotides, monomethoxytrityl (MMT)-protected 5' or (3')-amino-modified oligonucleotide intermediates are generated using the phosphoramidate monomer coupling method and detritylated as described in U.S. Patent No. 10,450,342. The 5' (or 3') MMT-protected amino group may be linked to the oligonucleotide through a linker group such as an alkyl phosphate group, and the MMT group may be removed from the oligonucleotide via solution-phase detritylation conducted at certain temperatures and pH. In certain embodiments, the detritylated oligonucleotide is then reacted with a conjugate group (e.g., a GalNAcs) to generate a conjugated oligonucleotide.

[0700] 2. Certain Cell-targeting Moieties

[0701] In certain embodiments, a conjugate moiety comprises or consists of a cell-targeting moiety. In certain embodiments, a cell-targeting moiety has affinity for a cell surface receptor on a cell. In certain embodiments, a cell-targeting moiety has affinity for a cell surface moiety on a cell. In certain embodiments, a cell-targeting moiety is capable of binding a cell surface receptor on a cell. In certain embodiments, a cell-targeting moiety is capable of binding a cell surface moiety on a cell. In certain embodiments, an oligomeric agent comprising a cell -targeting moiety is capable of being internalized by the cell when the cell-targeting moiety interacts with and / or binds a cell surface receptor and / or cell surface moiety. In certain embodiments, a cell surface receptor is not expressed ubiquitously (e.g., the cell surface receptor is undetectable in at least one tissue of a human subject), and a cell-targeting moiety selectively delivers an oligomeric agent, a modified oligonucleotide, or an oligomeric duplex to a tissue of interest or a cell of interest. By way of non-limiting example, the tissue of interest may be any one or more of brain, spinal cord, retina, heart, kidney, liver, lung, skeletal muscle, cardiac muscle, smooth muscle, adipose, white adipose, brown adipose, spleen, bone, intestine, colon, testes, breast, ovary, placenta, uterus, bladder, pancreas, pituitary, prostate, skin, adrenal gland, and thyroid. By way of non-limiting example, the cell of interest may be any one or more of a myocyte, adipocyte, hepatocyte, cardiomyocyte, vascular smooth muscle cell, endothelial cell, neuron, blood cell, macrophage, lymphocyte, cancer cell, and immune cell.

[0702] Asialoglycoprotein Receptor Ligands

[0703] In certain embodiments, a cell-targeting moiety has affinity for the hepatic asialoglycoprotein receptor (ASGP-R). In certain embodiments, the cell-targeting moiety comprises more than one ligand, and each ligand has affinity for the ASGP-R. In certain embodiments, each ligand is a carbohydrate. In certain embodiments, each ligand is N-acetyl galactosamine (GalNAc). In certain embodiments, the celltargeting moiety comprises three GalNAc ligands. In certain embodiments, the cell -targeting moiety is any one of those described in US 9,127,276.

[0704] GLP-1 Receptor Ligands

[0705] In certain embodiments, a cell -targeting moiety has affinity for a GEP-1 receptor. In certain embodiments, the cell-targeting moiety is any one of those described in US 2019 / 0134214.

[0706] GABA Transporter & Sortilin Receptor Ligands

[0707] In certain embodiments, a cell -targeting moiety has affinity for neurons. CHEMO 113W0

[0708] In certain embodiments, the cell-targeting moiety has affinity for a neurotransmitter receptor. In certain embodiments, a cell-targeting moiety has affinity for a Sortilin receptor. In certain embodiments, the cell-targeting moiety is any one of those described in WO 2021 / 236599.

[0709] In certain embodiments, the cell-targeting moiety has affinity for a neurotransmitter transporter. In certain embodiments, the cell targeting moiety targets a GABA transporter. See e.g., WO 2011 / 131693, WO 2014 / 064257.

[0710] Angiotensin II Type 1 Receptor Ligands

[0711] In certain embodiments, a cell -targeting moiety has affinity for an Angiotensin II Type I (AGTR1) receptor. In certain embodiments, the cell-targeting moiety is any one of those described in US 2022 / 0243210.

[0712] Integrin Receptor Ligands

[0713] In certain embodiments, a cell-targeting moiety has affinity for an integrin. In certain embodiments, the cell-targeting moiety has affinity for integrin avP3 and / or avP5. In certain embodiments, the celltargeting moiety is any one of those described in any of WO 2019 / 210200, WO 2019 / 210308. In certain embodiments, the cell-targeting moiety has affinity for integrin avP6. In certain embodiments, the celltargeting moiety is any one of those described in any of WO 2018 / 085415, WO 2019 / 089765, WO 2022 / 056269, WO 2022 / 056277, or WO 2022 / 056286.

[0714] Transferrin Receptor Ligands

[0715] In certain embodiments, a cell-targeting moiety has affinity for the type 1 transferrin receptor (TfRl; also known as CD71). In certain embodiments, a cell-targeting moiety comprises an anti-TfRl antibody or antigen-binding fragment thereof. In certain embodiments, the conjugate group comprises a protein or peptide capable of binding TfRl. In certain embodiments, the conjugate group comprises an aptamer capable of binding TfRl.

[0716] In certain embodiments, the anti-TfRl antibody or antigen-binding fragment thereof may be any known in the art including but not limited to those described in WO 1991 / 004753; WO 2013 / 103800; WO 2014 / 144060; WO 2016 / 081643; WO 2016 / 179257; WO 2016 / 207240; WO 2017 / 221883; WO 2018 / 129384; WO 2018 / 124121; WO 2019 / 151539; WO 2020 / 132584; WO 2020 / 028864; US 7,208,174; US 9,034,329; US 10,550,188; and US 11,512,136. In certain embodiments, a fragment of an anti-TfRl antibody is a F(ab')2, Fab, Fab', Fv, scFv, VHH, or VNAR. In certain embodiments, an antibody binds to TfRl through an engineered Fc domain rather than through the antigen-binding portion, as described in, e.g., US 2020 / 0223935.

[0717] In certain embodiments, the conjugate group comprises a protein or peptide capable of binding TfRl that does not include the antigen-binding fragment of an antibody. In certain embodiments, the protein or peptide capable of binding TfRl may be any known in the art including but not limited to those described in WO 2019 / 140050; WO 2020 / 037150; WO 2020 / 124032; WO 2022 / 026555; WO 2023 / 027125; WO 2023 / 022234; and US 10,138,483. In certain embodiments, the peptide is a cyclic peptide, as described in WO 2021 / 167107. In certain embodiments, the peptide is a bicyclic peptide known as a ‘bicycle ligand’ selected from those described in WO 2022 / 101633 and WO 2023 / 056388. In certain embodiments, the CHEMO 113W0 conjugate group comprises an aptamer capable of binding TfRl. In certain embodiments, the aptamer capable of binding TfRl may be any known in the art including but not limited to those described in WO 2013 / 163303; WO 2019 / 033051; and WO 2020 / 245198.

[0718] 3. Conjugate Linkers

[0719] In certain embodiments, oligomeric agents comprise an oligonucleotide and a conjugate group, wherein the conjugate group consists of a conjugate moiety and a conjugate linker. The conjugate linker links the conjugate moiety to the oligonucleotide. In certain embodiments, the conjugate linker is a single chemical bond (z.e., the conjugate moiety is attached directly to an oligonucleotide through a single bond). In certain embodiments, the conjugate linker comprises one or more atoms. In certain embodiments, the conjugate linker comprises a chain structure, such as a hydrocarbyl chain, or an oligomer of repeating units such as ethylene glycol, nucleosides, or amino acid units. In certain embodiments, the oligonucleotide is a modified oligonucleotide.

[0720] In certain embodiments, a conjugate linker comprises one or more groups selected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol, ether, thioether, and hydroxylamino. In certain such embodiments, the conjugate linker comprises one or more groups selected from alkyl, amino, oxo, amide, and ether groups. In certain embodiments, the conjugate linker comprises one or more groups selected from alkyl and amide groups. In certain embodiments, the conjugate linker comprises one or more groups selected from alkyl and ether groups. In certain embodiments, the conjugate linker comprises at least one phosphorus moiety. In certain embodiments, the conjugate linker comprises at least one phosphate group. In certain embodiments, the conjugate linker includes at least one neutral linking group.

[0721] In certain embodiments, conjugate linkers, including the conjugate linkers described herein, are bifunctional linking moieties, e.g., those known in the art to be useful for attaching conjugate moieties to parent compounds, such as the oligonucleotides provided herein. In general, a bifunctional linking moiety comprises at least two functional groups. One of the functional groups is selected to react with a particular site on a parent compound and the other is selected to react with a conjugate moiety. Examples of functional groups used in a bifunctional linking moiety include but are not limited to electrophiles for reacting with nucleophilic groups and nucleophiles for reacting with electrophilic groups. In certain embodiments, bifunctional linking moieties comprise one or more groups selected from amino, hydroxyl, carboxylic acid, thiol, alkyl, alkenyl, and alkynyl.

[0722] In certain embodiments, conjugate linkers comprise chemical groups that are formed upon a reaction between a first functional group and a second functional group. In certain embodiments, an oligonucleotide (e.g., a modified oligonucleotide) is attached to the first functional group during synthesis, and a conjugate moiety is attached to a second functional group during synthesis. Then, the two compounds are mixed under specific conditions to yield the oligonucleotide covalently linked to the conjugate moiety. Such reactions that are compatible with both oligonucleotide and peptide chemistry have been previously described and are often called “bioconjugation” reactions. These reactions include strain-promoted azidealkyne cycloaddition (SPAAC), copper-catalyzed click reaction (CuAAC), active ester conjugation to an CHEMO 113W0 amino modified oligonucleotide, maleimide -thiol Michael addition, ketol / hydroxylamine ligation, the Staudinger ligation, reductive amination, thioether formation, disulfide formation, reductive alkylation, catalyst-free N-arylation, sulfur fluoride exchange click reaction (SuFEx), and inverse demand Diels-Alder reaction. Certain such reactions are described in, e.g., Jbara, M., etal. Angew. Chem. Int. Ed. 2021, 60 (21), 12109-12115; Dong, J., et al. Angew. Chem. Int. Ed. 2014, 53 (36), 9430-9448.4; Zhang, C. et al. Angew. Chem. Int. Ed. Engl. 2019, 58 (15), 4810-4839; Walsh, S. J., et al. Chem. Soc. Rev., 2021, 50, 1305-1353; Tiefenbrunn, T. et al. Biopolymers, 2010, 94 (1), 95-106; Drake, P. M., et al. Bioconjug. Chem. 2014, 25 (7), 1331-1341; Bode, J. W„ Acc. Chem. Res., 2017, 50 (9), 2104-2115; Magano, J., etal. Org. Proc. Res. Dev. 2014, 18, 142-151; McKay, C. S. and Finn, M. G. Chem. Biol. 2014, 21 (9), 1075-101; Christy, M. P. et al. Org. Lett. 2020, 22, 2365; Ren, H., et al. Angew. Chem. Int. Ed. Engl. 2009, 48, 9658-9662; Rohrbacher, F. et al., Helv. Chim. Acta. 2018, 101 (5), el800039; Baalmaan, M., et al. Angew. Chem. Int. Ed. 2020, 59 (31), 12885-12893; Lang, K„ et al. J. Am. Chem. Soc. 2014, 9 (1), 16-20; Nair, D. P„ et al. Chem. Mater. 2013 26 (1), 724-744; Kalia, J. and Raines, R. T. Angew. Chem. Int. Ed., 2008, 47, 7523- 7526.

[0723] Examples of conjugate linkers include but are not limited to pyrrolidine, 8-amino-3,6- dioxaoctanoic acid (ADO), succinimidyl 4-(N-maleimidomethyl) cyclohexane- 1 -carboxylate (SMCC) and 6-aminohexanoic acid (AHEX or AHA). Other conjugate linkers include but are not limited to substituted or unsubstituted Ci-Cw alkyl, substituted or unsubstituted C2-C10 alkenyl or substituted or unsubstituted C2- C10 alkynyl, wherein a nonlimiting list of substituent groups includes hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro, thiol, thioalkoxy, halogen, alkyl, aryl, alkenyl and alkynyl.

[0724] In certain embodiments, it is desirable for a conjugate moiety to be cleaved from the oligonucleotide . For example, in certain circumstances oligomeric agents comprising a particular conjugate moiety are better taken up by a particular cell type, but once the oligomeric agent has been taken up, it is desirable that the conjugate moiety be cleaved to release the unconjugated oligonucleotide or oligomeric duplex. Thus, certain conjugate linkers may comprise one or more cleavable moieties. In certain embodiments, a cleavable moiety is a cleavable bond. In certain embodiments, a cleavable moiety is a group of atoms comprising at least one cleavable bond. In certain embodiments, a cleavable moiety comprises a group of atoms having one, two, three, four, or more than four cleavable bonds. In certain embodiments, a cleavable moiety is selectively cleaved inside a cell or subcellular compartment, such as a lysosome. In certain embodiments, a cleavable moiety is selectively cleaved by endogenous enzymes, such as nucleases.

[0725] A “cleavable moiety” means a group of atoms comprising at least one bond that is cleaved under physiological conditions, e.g., in a cell and / or a subject. For example, a cleavable moiety cleaved inside a cell or sub-cellular compartment, such as an endosome or lysosome. A cleavable moiety may be cleaved by endogenous enzymes, such as nucleases.

[0726] In certain embodiments, a cleavable bond is selected from an amide, an ester, an ether, one or both esters of a phosphodiester, a phosphate ester, a carbamate, or a disulfide. In certain embodiments, a cleavable bond is one or both of the esters of a phosphodiester. In certain embodiments, a cleavable moiety comprises a phosphate or phosphodiester. In certain embodiments, the cleavable moiety is a phosphodiester CHEMO 113W0 linkage between an oligonucleotide and a conjugate moiety.

[0727] In certain embodiments, a cleavable moiety may be part of the oligonucleotide and comprises or consists of one or more linked nucleosides. In certain such embodiments, the one or more linked nucleosides are linked to one another and / or to the remainder of the oligonucleotide through cleavable bonds. In certain embodiments, such cleavable bonds are unmodified phosphodiester bonds. In certain embodiments, a cleavable moiety is 2'-deoxynucleoside that is either the 3' or 5 '-terminal nucleoside of an oligonucleotide linked by a phosphodiester intemucleoside linkage to an adjacent nucleoside of the oligonucleotide and covalently attached to the conjugate linker or conjugate moiety by a phosphodiester or phosphorothioate linkage. In certain such embodiments, the cleavable moiety comprises 2'-deoxyadenosine.

[0728] In certain embodiments, oligomeric agents described herein comprise an oligonucleotide linked to a conjugate moiety by a conjugate linker, wherein the oligonucleotide is attached to the conjugate moiety using Click chemistry known in the art. Compounds have been prepared using Click chemistry wherein alkynyl phosphonate intemucleoside linkages on an oligonucleotide attached to a solid support are converted into the 1,2,3-triazolylphosphonate intemucleoside linkages and then cleaved from the solid support (Krishna, H. et al. J. Am. Chem. Soc. 2012, 134(28), 11618-11631). Additional conjugate linkers suitable for oligonucleotide conjugates are prepared by Click chemistry described in “Click Chemistry for Biotechnology and Materials Science” Ed. Joerg Lahann, Wiley 2009. Further examples of linking chemistry include an inverse electron demand Diels-Alder reaction, e.g., as described in Argamunt et al., J. Org. Chem. 2020, 85, 10, 6593-6604, Sarrett et al. , Nat. Protocols 2021, 16, 3348-3381; Handula et al. , Molecules, 2021, 26 (15), 4640, Wiessler et al. , Int. J. Med. Set. 2010, 7 (1), 19-28; copper-catalyzed azidealkyne cycloaddition (CuAAC) see, e.g., S. I. Presolski, et al., J. Am. Chem. Soc. 2010, 132, 14570-14576; D. Soriano Del Amo, et al., J. Am. Chem. Soc., 2010, 132, 16893-16899; Staudinger reaction, see, e.g., Saxon and C. R. Bertozzi, Science, 2000, 287, 2007-2010; B. L. Nilsson, et al., Org. Lett., 2000, 2, 1939— 1941, E. Saxon, etal., Org. Lett., 2000, 2, 2141-2143; formation of hydrazones and oximes, see, e.g., J. Y. Axup, et al., Proc. Natl. Acad. Sci. U. S. A., 2012, 109, 16101-16106; photoclick reactions, see, e.g., W. Song, etal., Angew. Chem., Int. Ed., 2008, 47, 2832-2835, A. Hemer and Q. Lin, Top. Curr. Chem., 2016, 374, 1; strain-promoted alkyne-nitrone cycloaddition (SPANC) reactions, see, e.g., D. A. MacKenzie, et al., Curr. Opin. Chem. Biol., 2014, 21, 81-88; transition metal catalyzed cross coupling, see, e.g., M. Chalker, etal., J. Am. Chem. Soc., 2009, 131, 16346-16347; nucleophilic additions, in particular, of a thiol to a maleimide, see, e.g., Kang et al., Chem. Sci., 2021, 12, 13613-13647, Bemardim et al., Nat. Comm. 2016, 7, 13128, Jain et al., Pharm. Res. 2015, 32 (11), 3526-3540.

[0729] Synthetic methods describing preparation of the above starting materials and intermediates can be found in one or more of the following: Agard, N. J., et al. “A Strain-Promoted [3 + 2] Azide-Alkyne Cycloaddition for Covalent Modification of Biomolecules in Living Systems.” J. Am. Chem. Soc. 2004, 126, 15046- 15047; Lang, K., and Chin, J. W. “Biorthogonal Reactions for Labeling Proteins.” ACS Chem. Biol. 2014, 9 (1), 16-20; Nair, D. P. et al. “The Thiol-Michael Addition Click Reaction: A Powerful and Widely Used Tool in Materials Chemistry.” Chem. Mater. 2013 26 (1), 724-744; WO2011 / 136645; Kolmel, D. K. and Kool, E. T. “Oximes and Hydrazones in Bioconjugation: Mechanism and Catalysis.” Chem. Rev. 2017, 117, 10358-10376; Wang, J. et al. “Polyfluorophenyl Ester-Terminated Homobifunctional CrossLinkers for Protein Conjugation.” Synlett, 2017, 28 (15), 1934-1938; Kishimoto, S. et al. “Site-Specific Chemical Conjugation of Antibodies by Using Affinity Peptide for the Development of Therapeutic Antibody Format,” Bioconjugate Chem., 2019, 30 (3), 698-702, Wu and Devaraj, “Inverse Electron- Demand Diels-Alder Bioorthogonal Reactions,” Top. Curr. Chem. 2016, 374, 3, Oliveira et al., “Inverse electron demand Diels-Alder reactions in chemical biology,” Chem. Soc. Rev., 2017, 46, 4895-4950.

[0730] In certain embodiments, the linker is prepared by reaction of a first reactive moiety with a second reactive moiety, wherein the first reactive moiety is attached to the oligonucleotide and the second reactive moiety is attached to the conjugate moiety, or a precursor thereof. In certain embodiments, the linker is prepared by reaction of a dipolarophile (e.g. , a triple bonded moiety such as an alkyne or nitrile) with a 1,3-dipole (e.g., an azide, a nitrone, an isocyanate, or a thioisocyanate): wherein each Q is independently a carbon atom or a heteroatom, for example, one of X and Y is attached to a cargo such as an oligonucleotide, and the other of X and Y is attached to a conjugate moiety such as a transferrin-receptor binding moiety or a half-life extension moiety. The linker thus prepared may comprise a five-membered unsaturated heterocyclic ring such as a triazole.

[0731] In certain embodiments, the linker is prepared by reaction of a dieneophile (e.g., an electron rich double bond such as a furan or derivative thereof) with an electron poor diene (e.g., a tetrazine): wherein each Q is independently a carbon atom or a heteroatom, for example, one of X and Y is attached to a cargo such as an oligonucleotide, and the other of X and Y is attached to a conjugate moiety such as a transferrin-receptor binding moiety or a half-life extension moiety. The linker thus prepared may comprise a six-membered unsaturated heterocyclic ring such as a dihydropyrazine.

[0732] In certain embodiments, the linker is prepared by reaction of a nucleophile (e.g., a thiol or amine) with an electrophile (e.g., an electron-poor carbonyl or carbonyl-conjugated alkene or alkyne): CHEMO 113W0 wherein each Q is independently a carbon atom or a heteroatom, for example, one of X and Y is attached to a cargo such as an oligonucleotide, and the other of X and Y is attached to a conjugate moiety such as a transferrin-receptor binding moiety or a half-life extension moiety. The linker thus prepared may comprise a thioether, hydrazone, oxime, or amide.

[0733] Each of the first reactive moiety and the second reactive moiety may attach at any suitable position of the modified oligonucleotide and the conjugate moiety. For example, at a 3’- or 5 ’-terminal position, or at a 2’-position of a furanosyl sugar moiety, to a nucleobase, to an intemucleoside linkage, or to a sugar surrogate. In certain embodiments, the linker attaches at the 5 ’-terminal hydroxyl group of a modified oligonucleotide.

[0734] In certain embodiments, a linker has Formula L:

[0735] [(RMl)-(RU)qi-(RM2)]q2

[0736] L wherein each RM1 and RM2 is independently absent or a functional group derived from conjugation of a reactive moiety, for example wherein RM1 and RM2 are independently selected from an amino acid, O, NH, NRa2, S-S, O-NH, O-NRa2, NH-NH, NH-NRa2, NRa2-NRa2, Ci6haloalkylene, C2-6 alkenylene, C2-6 alkynylene, C3-10 cycloalkylene, Ce-io arylene, heteroarylene, heterocyclylene, Ci-e alkylene-O-, C2-6 alkenylene-O-, C2-6 alkynylene-O-, C1-6 alkylene-C(O)O, Ci-e alkylene-NH, Ci.6 alkylene-NRa2, C1-6 alkylene-C(O)NH, Ci-6 alkylene-C(O)NRa2, C(O), C(O)O, C(O)NH, C(O)NRa2, P(O)(OH), P(O)(NRa2), P(O)(SH), P(S)(NRa2), P(Ra2), P(NRa2), S, S(O)2, S(O), SRa2, and S(NRa2); each Ra2is independently selected from C(O)Ra3, C(O)ORa3, C(O)NHRa3, C(O)N(Ci.4alkyl)Ra3, SRa3, S(O)2Ra3, S(O)Ra3, Ci.6alkyl, C2.6alkenyl, C2-6alkynyl, C3-10 cycloalkyl, Ce-io aryl, heteroaryl, and heterocyclyl; each Ra3is independently hydrogen, OH, C1-6 alkyl, Ci-ehaloalkyl, C3-10 cycloalkyl, Ce-io aryl, heteroaryl, or heterocyclyl; and each RU is a repeating unit independently selected from C1-6 alkylene, C1-6 haloalkylene, C1-6 heteroalkylene, C2-6 alkenylene, C2-6 alkynylene, C3-10 cycloalkylene, Ce-io arylene, heteroarylene, heterocyclylene, Ci-e alkylene-O-, C2-6 alkenylene-O-, C2-6 alkynylene-O-, C1-6 alkylene-C(O)O, Ci.e alkylene-NH-, Ci-e alkylene-NRa2-, Ci.6 alkylene-C(O)NH, Ci-e alkylene-C(O)NRa2, a nucleotide, a saccharide, and an amino acid; and each RU is optionally substituted one with one, two, three, four, or five groups independently selected from halo (for example, fluoro), cyano, azido, OH, ORa2, NO2, NH2, NHRa2, N(Ra2)2, Ci-e alkyl, Ci-ehaloalkyl, C2-6 alkenyl, C2-e alkynyl, C3-10 cycloalkyl, Ce-io aryl, heteroaryl, heterocyclyl, C1-6 alkylene-ORa2, C2-6 alkenylene -ORa2, C2-6 alkynylene -ORa2, Ci- e alkylene-NH2, C1-6 alkylene-NHRa2, C1-6 alkylene -N(Ra2)2, C(O)Ra3, C(O)ORa3, C(O)NHRa3, C(O)N(Ci.4alkyl)Ra3, SRa3, S(O)2Ra3, S(O)Ra3, NHC(O)Ra3, N(CI4alkyl)C(O)Ra3, NHS(O)Ra3, N(Ci.4alkyl)S(O)Ra3, NHS(O)2Ra3, and N(Ci.4alkyl)S(O)2Ra3; each Ra2is independently selected from C(O)Ra3, C(O)ORa3, C(O)NHRa3, C(O)N(Ci.4alkyl)Ra3, SRa3, S(O)2Ra3, S(O)Ra3, C1-6 alkyl, C2-6 a lkenyl, C2-e alkynyl, C3-10 cycloalkyl, Ce-io aryl, heteroaryl, and CHEMO 113W0 heterocyclyl; each Ra3is independently hydrogen, OH, C1-6 alkyl, Ci-ehaloalkyl, C3- 10 cycloalkyl, Ce-io aryl, heteroaryl, or heterocyclyl; or RU is a hydrophilic moiety; and wherein ql is 0-30 and q2 is 1-20.

[0737] In certain embodiments, an amino acid has a structure (amino acid) wherein SC is an amino acid side chain, optionally a side chain found in a natural amino acid, and Ra4is

[0738] H or Ra2as defined with respected to Formula L. In certain embodiments, SC is H or CH2OH. In certain embodiments, Ra2is methyl.

[0739] In certain embodiments, a saccharide has a structure (saccharide) wherein Q1is OH, NH2, or NHAc and wherein two of Q2are H and one of Q2attaches to the remainder of the linker.

[0740] In certain embodiments, each R 1 and RM2 is independently selected from phosphate, diphosphate, triphosphate, phosphorothioate, phosphorodithioate, phosphorothiolate; phosphoramidates, alkylphosphonates, CH2-C(O)NH, O, NH, triazole, amide, carbonate, carbamate, urea, N- hydroxy succinimide, oxime, hydrazone, disulfide, heteroaryl, heterocyclyl,

[0741] (amino acid), .

[0742] In certain embodiments, each R 1 and RM2 is independently selected from P(O)(OH), O, CH2- C(O)NH , C(O)NH, or NH, and RU is ethylene glycol or CH2optionally substituted with C(O)OH.

[0743] In certain embodiments, each RU is a hydrophilic moiety independently selected from y g y , (amino acid), (sar), and CH2, wherein SC is an amino acid side chain, optionally a side chain found in a natural amino acid.

[0744] In certain embodiments, each R 1 and RM2 is independently selected from amino acid, C(O)NH, and heteroaryl, and RU is ethylene glycol or optionally substituted CH2. CHEMO 113W0

[0745] In certain embodiments, each RU is ethylene glycol or sar. In certain embodiments, each RU is ethylene glycol and ql is 2-6. In certain embodiments, each RU is sar and ql is 2-6. In certain embodiments, each RU is CH2 and ql is 1-6.

[0746] In certain embodiments, RU is an amino acid, and SC is H or CH2OH. In certain embodiments, RU comprises glycine and serine. In certain embodiments, (RU)qiis (G;S),i or (G4S)r2, wherein G is glycine and S is serine, and rl and r2 are 1-5. In certain embodiments, (RU)qiis GGGS. In certain embodiments, each RU is an independently selected amino acid and (RU)qiis an amino acid sequence disclosed in US Patent No. 7,612,181. In certain embodiments, (RU)qiis ASTKGP or TVAAPSVFIFPP. In certain embodiments, (RU)qiis EPKSCDG4S, EPKSCD(G4S)2, or EPKSCD(G4S)3. In certain embodiments, (RU)qiis Val-Cit, Gly-Val-Cit, or Gly-Gly-Gly.

[0747] Certain Click Linkers

[0748] In certain embodiments, a Click reaction is used to form a linker by reacting: with an oligonucleotide having a terminal amine, including but not limited to the following compound: wherein Y is the oligonucleotide, to yield: which is reacted with a conjugate moiety having an azide to yield:

[0749] In certain embodiments, a linker is prepared from the following compound: In certain embodiments, a Click reaction is used to form a linker by reacting: solution together with an oligonucleotide having a terminal amine, including but not limited to the following compound: which is reacted with an azide to yield:

[0750] In certain embodiments, a linker is prepared from the following compound:

[0751] In certain embodiments, a Click reaction is used to link a conjugate moiety and an oligonucleotide by reacting: wherein one of Y and Y’ is attached to the conjugate moiety and X is attached to the oligonucleotide, to yield:

[0752] In certain embodiments, a linker comprises:

[0753]

[0754] In certain embodiments, a linker comprises: wherein each of X and Y is an oligonucleotide or a conjugate moiety. CHEMO 113W0

[0755] Synthetic methods describing preparation of the above starting materials and intermediates can be found in one or more of the following: Agard, N. J., et al. “A Strain-Promoted [3 + 2] Azide-Alkyne Cycloaddition for Covalent Modification of Biomolecules in Living Systems.” J. Am. Chem. Soc. 2004, 126, 15046- 15047; Lang, K., and Chin, J. W. “Biorthogonal Reactions for Labeling Proteins.” ACS Chem. Biol. 2014, 9 (1), 16-20; Nair, D. P. et al. “The Thiol-Michael Addition Click Reaction: A Powerful and Widely Used Tool in Materials Chemistry.” Chem. Mater. 2013 26 (1), 724-744;

[0756] WO2011 / 136645; Kolmel, D. K. and Kool, E. T. “Oximes and Hydrazones in Bioconjugation: Mechanism and Catalysis.” Chem. Rev. 2017, 117, 10358-10376; Wang, J. et al. “Polyfluorophenyl Ester-Terminated Homobifunctional Cross-Linkers for Protein Conjugation.” Synlett, 2017, 28 (15), 1934-1938; Kishimoto, S. et al. “Site-Specific Chemical Conjugation of Antibodies by Using Affinity Peptide for the Development of Therapeutic Antibody Lormat,” Bioconjugate Chem., 2019, 30 (3), 698- 702, Wu and Devaraj, “Inverse Electron-Demand Diels-Alder Bioorthogonal Reactions,” Topics in Current Chemistry 2016, 374, 3, Oliveira et al., “Inverse electron demand Diels-Alder reactions in chemical biology,” Chem. Soc. Rev., 2017, 46, 4895-4950.

[0757] H. Terminal Groups

[0758] In certain embodiments, provided herein are oligomeric agents comprising one or more modified oligonucleotides and one or more terminal groups. As used herein, “terminal group” means a group of atoms that is covalently linked to a terminus of an oligonucleotide. Examples of a terminal group include, but are not limited to, a capping group, a phosphate moiety, a stabilized phosphate group, and a protecting group. In certain embodiments, one or more terminal groups is attached to either or both ends of an oligonucleotide. In certain embodiments, one or more terminal groups is attached at the 3 '-end and / or at the 5 '-end of the oligonucleotide. In certain embodiments, one or more terminal groups is attached at the 3'- end of the oligonucleotide.

[0759] In certain embodiments, an oligonucleotide is linked to a terminal group comprising a stabilized 5 '-phosphate. In certain embodiments, in an oligomeric duplex provided herein, a terminal group comprising a stabilized phosphate moiety is attached at the 5 '-end of the first modified oligonucleotide. The stabilized phosphate moiety results in stabilization of a 5 '-phosphate moiety of the 5 '-terminal nucleoside of an oligonucleotide, relative to the stability of an unmodified 5 '-phosphate of an unmodified nucleoside under biologic conditions. Such stabilization of a 5 '-phosphate group includes but is not limited to resistance to removal by phosphatases. Stabilized phosphate moieties, but are not limited to 5 '-phosphonates, including, but not limited to 5'-vinylphosphonate, 5'-methylphosphonate, and 5 '-cyclopropyl phosphonate. In certain embodiments, the stabilized phosphate moiety is a cyclopropyl phosphonate or an f / ’.j-vinyl phosphonate.

[0760] In certain embodiments, the oligomeric agent comprises a capping group. In certain embodiments, the capping group is an abasic nucleoside. CHEMO 113W0

[0761] I. Target Nucleic Acids

[0762] In certain embodiments, oligomeric agents comprise or consist of a modified oligonucleotide comprising a targeting region that is complementary to an equal-length target region of a target nucleic acid.

[0763] In certain embodiments, the target nucleic acid is an endogenous RNA molecule. In certain embodiments, the nucleic acid encodes a protein. In certain such embodiments, the nucleic acid is selected from: a mature mRNA and a pre-mRNA, including intronic, exonic, and untranslated regions. In certain embodiments, the RNA is a mature mRNA. In certain embodiments, the nucleic acid is a pre-mRNA.

[0764] II. Methods and Uses

[0765] A. Antisense Activity

[0766] In certain embodiments, oligomeric agents provided herein comprise an antisense oligonucleotide that is capable of hybridizing to a target nucleic acid, resulting in at least one antisense activity; such oligomeric agents are antisense agents.

[0767] In certain antisense activities, hybridization of an antisense oligonucleotide to a target nucleic acid results in recruitment of a protein, e.g., RNase H or Argonaute, that cleaves the target nucleic acid. Certain antisense agents result in RNase H mediated cleavage of the target nucleic acid. RNase H is a cellular endonuclease that cleaves the RNA strand of an RNA:DNA duplex. The DNA in such an RNA:DNA duplex need not be unmodified DNA. In certain embodiments, oligomeric agents are antisense agents that are sufficiently “DNA-like” to elicit RNase H activity. In certain embodiments, one or more non-DNA-like nucleosides in the antisense agent are tolerated and RNase H activity is retained. In certain embodiments, such antisense agents reduce expression of or reduce the amount or activity of a target nucleic acid by 25% or more in the standard in vitro assay.

[0768] In certain antisense activities, an antisense oligonucleotide is loaded into an RNA-induced silencing complex (RISC), ultimately resulting in cleavage of the target nucleic acid. For example, certain antisense oligonucleotides result in cleavage of the target nucleic acid by Argonaute. Antisense agents that comprise an antisense oligonucleotide that is loaded into RISC are RNAi agents. RNAi agents may be doublestranded (siRNA or dsRNAi) or single-stranded (ssRNA). In certain embodiments, RNAi agents are capable of RISC-mediated modulation of a target nucleic acid in a cell. In certain embodiments, such RNAi agents reduce or the expression of or reduce the amount or activity of a target nucleic acid by 25% or more in the standard in vitro assay. In certain embodiments, RNAi agents selectively affect one or more target nucleic acid. Such RNAi agents comprise a modified oligonucleotide having a nucleobase sequence that hybridizes to one or more target nucleic acid, resulting in one or more desired antisense activity. In certain embodiments, an RNAi agent comprises a modified oligonucleotide that does not hybridize to one or more non-target nucleic acid or does not hybridize to one or more non-target nucleic acid in such a way that results in significant undesired antisense activity.

[0769] In certain embodiments, hybridization of an antisense oligonucleotide to a target nucleic acid does not result in recruitment of a protein that cleaves that target nucleic acid. In certain embodiments, CHEMO 113W0 hybridization of the antisense oligonucleotide to the target nucleic acid results in alteration of splicing of the target nucleic acid. In certain embodiments, hybridization of an antisense oligonucleotide to a target nucleic acid results in exon inclusion or exon exclusion. In certain embodiments, hybridization of an antisense oligonucleotide to a target nucleic acid results in retained intron exclusion.

[0770] In certain embodiments, hybridization of an antisense compound to a target nucleic acid results in inhibition of a binding interaction between the target nucleic acid and a protein or other nucleic acid (e.g., miRNA, IncRNA, sncRNA). In certain embodiments, hybridization of an antisense compound to a target nucleic acid results in modulation of translation of the target nucleic acid. In certain embodiments, hybridization of an oligomeric agent to a target nucleic acid results in an increase in the amount or activity of a target nucleic acid. In certain embodiments, hybridization of an antisense oligonucleotide to a target nucleic acid results in increased translation of the target nucleic acid. In certain embodiments, hybridization of an antisense oligonucleotide to a target nucleic acid results in reduced translation of the target nucleic acid.

[0771] Antisense activities may be observed directly or indirectly. In certain embodiments, observation or detection of an antisense activity involves observation or detection of a change in an amount of a target nucleic acid or protein encoded by such target nucleic acid, a change in the ratio of splice variants of a nucleic acid or protein and / or a phenotypic change in a cell or animal.

[0772] B. Treatment, Prophylaxis

[0773] In certain embodiments, provided herein are methods of modulating expression, RNA levels, and / or protein levels and / or activity, in a subject having, or at risk of having, a disease, disorder, condition or injury. In certain embodiments, the disease, disorder, condition, or injury is of the CNS of a subject. In certain embodiments, administering such an oligomeric agent modulates target expression, RNA levels and / or protein levels in the CNS of the subject. In some instances, such an oligomeric agent is administered parenterally. In some instances, an oligomeric agent is administered intravenously, subcutaneously, intramuscularly, or intrathecally. In some instances, an oligomeric agent is administered subcutaneously. In some instances, an oligomeric agent is administered intrathecally. In certain embodiments, the detectable amount of the target RNA may be reduced by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% compared to a control.

[0774] In certain embodiments, an oligomeric agent described herein has improved tolerability or reduced off-target effect compared to an oligomeric agent in which neither ofWl and / or W2 comprises a 2’-deoxynucleoside; and / or G consists of no more than 10 linked nucleosides; and / or the modified oligonucleotide does not comprise at least one mesyl phosphoramidate intemucleoside linkage. In certain embodiments, improved tolerability is determined by BJAB activation assay. In certain embodiments, reduced off-target effect is determined by IC50 for knockdown of one or more off-target genes. In certain embodiments, reduced off-target effect is determined by number of differentially expressed off-target gene responders, optionally wherein the responders are critical responders. CHEMO 113W0

[0775] III. Pharmaceutical Compositions

[0776] In certain embodiments, a pharmaceutical composition comprises a pharmaceutically acceptable diluent or carrier. In certain embodiments, a pharmaceutical composition comprises or consists of a sterile saline solution and one or more oligomeric agent. In certain embodiments, the sterile saline is pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition comprises or consists of one or more oligomeric agent (e.g. , a modified oligonucleotide or oligomeric duplex) provided herein and sterile water. In certain embodiments, the sterile water is pharmaceutical grade water. In certain embodiments, a pharmaceutical composition comprises or consists of one or more oligomeric agent (e.g., a modified oligonucleotide or oligomeric duplex) provided herein and phosphate-buffered saline (PBS). In certain embodiments, sterile PBS is pharmaceutical grade PBS. In certain embodiments, a pharmaceutical composition comprises or consists of one or more oligomeric agent (e.g., a modified oligonucleotide or oligomeric duplex) provided herein and artificial cerebrospinal fluid (“artificial CSF” or “aCSF”). In certain embodiments, the artificial cerebrospinal fluid is pharmaceutical grade artificial cerebrospinal fluid.

[0777] In certain embodiments, aCSF comprises one or more of sodium chloride, potassium chloride, sodium dihydrogen phosphate dihydrate, sodium phosphate dibasic anhydrous, calcium chloride dihydrate, and magnesium chloride hexahydrate. In certain embodiments, the pH of an aCSF solution is modulated with a suitable pH-adjusting agent, for example, with acids such as hydrochloric acid and alkalis such as sodium hydroxide, to a range of from about 7.1-7.3, or to about 7.2.

[0778] In certain embodiments, pharmaceutical compositions comprise one or more oligomeric agent and one or more excipients. In certain embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose and polyvinylpyrrolidone.

[0779] In certain embodiments, pharmaceutical compositions comprising an oligomeric agent encompass any pharmaceutically acceptable salts of the oligomeric agent, esters of the oligomeric agent, or salts of such esters. As used herein “pharmaceutically acceptable salts” means physiologically and pharmaceutically acceptable salts of compounds which do not impart undesired toxicological effects thereto. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium, potassium, calcium, and magnesium salts. In certain embodiments, pharmaceutical compositions comprising an oligomeric agent comprising or consisting of one or more modified oligonucleotide, upon administration to a subject, including a human, are capable of providing (directly or indirectly) the biologically active metabolite or residue thereof.

[0780] Under certain conditions, certain compounds disclosed herein act as acids. Although such compounds may be drawn or described in protonated (free acid) form or ionized and in association with a cation (salt) form, aqueous solutions of such compounds exist in equilibrium among such forms. For example, a phosphodiester linkage of an oligonucleotide in aqueous solution exists in equilibrium among free acid, anion and salt forms. Unless otherwise indicated, compounds described herein are intended to include all such forms. Moreover, certain oligonucleotides have several such linkages, each of which is in CHEMO 113W0 equilibrium. Thus, oligonucleotides in solution exist in an ensemble of forms at multiple positions all at equilibrium. The term “oligonucleotide” herein is intended to include all such forms.

[0781] Lipid moieties have been used in nucleic acid therapies in a variety of methods. In certain methods, a nucleic acid, such as an oligomeric agent comprising a modified oligonucleotide, is introduced into preformed liposomes or lipoplexes made of mixtures of cationic lipids and neutral lipids. In certain methods, nucleic acid complexes with mono- or poly-cationic lipids are formed without the presence of a neutral lipid. In certain embodiments, a lipid moiety is selected to increase distribution of an oligomeric agent to a particular cell or tissue . In certain embodiments, a lipid moiety is selected to increase distribution of an oligomeric agent to fat tissue. In certain embodiments, a lipid moiety is selected to increase distribution of an oligomeric agent to muscle tissue.

[0782] In certain embodiments, pharmaceutical compositions comprise a delivery system. Examples of delivery systems include, but are not limited to, lipid nanoparticles, liposomes, and emulsions. Certain delivery systems are useful for preparing certain pharmaceutical compositions including those comprising hydrophobic compounds. In certain embodiments, certain organic solvents such as dimethylsulfoxide are used.

[0783] Nonlimiting disclosure and incorporation by reference

[0784] Each of the literature and patent publications listed herein is incorporated by reference in its entirety.

[0785] While certain compounds, compositions, and methods have been described herein with specificity in accordance with certain embodiments, the following examples serve only to illustrate the compounds described herein and are not intended to limit the same. Each of the references, GenBank accession numbers, ENSEMBL identifiers, and the like recited in the present application, is incorporated herein by reference in its entirety.

[0786] The sequence listing accompanying this filing identifies each nucleic acid sequence as either “RNA” or “DNA” as required; however, one of skill in the art will readily appreciate that designation of “RNA” or “DNA” to describe modified oligonucleotides is, in certain instances, arbitrary. For example, an oligonucleotide comprising a nucleoside comprising a 2 ’-OH sugar moiety and a thymine base could be described as a DNA having a modified sugar (i.e., 2’-OH in place of one 2’-H of DNA) or as an RNA having a modified base (i.e., thymine (5-methyl uracil) in place of an uracil of RNA); and certain nucleic acid compounds described herein comprise one or more nucleosides comprising modified sugar moieties having 2’-substituent(s) that are neither OH nor H. One of skill in the art will readily appreciate that labeling such nucleic acid compounds “RNA” or “DNA” does not alter or limit the description of such nucleic acid compounds.

[0787] Herein, the description of compounds as having “the nucleobase sequence of’ a SEQ ID NO. describes only the nucleobase sequence. Accordingly, absent additional description, such description of compounds by reference to a nucleobase sequence of a SEQ ID NO. does not limit sugar or intemucleoside linkage modifications or presence or absence of additional substituents such as a CHEMO 113W0 conjugate group. Further, absent additional description, the nucleobases of a compound “having the nucleobase sequence of’ a SEQ ID NO. include such compounds having modified forms of the identified nucleobases as described herein.

[0788] Herein, the description of compounds by chemical notation (subscripts and / or superscripts to indicate chemical modifications) include each noted modification. Chemical notation includes the indicated nucleobases and sugar modifications in the 5’ to 3’ direction. For example, the chemical notation of “AesTkomCezGdsCd” indicates a compound wherein the first nucleoside comprises a 2’ -MOE sugar moiety (indicated by the “e” subscript) and an unmodified adenine nucleobase linked to the second nucleoside via a 3’-phosphorothioate linkage (indicated by the “s” subscript); the second nucleoside comprises a cEt sugar moiety (indicated by the “k” subscript) and an unmodified thymine nucleobase linked to the third nucleoside via a phosphodiester linkage (indicated by the “o” subscript); the third nucleoside comprises a 2’-MOE sugar moiety and a 5-methyl modified cytosine nucleobase (indicated by the “m” superscript) linked to the fourth nucleoside via a mesyl phosphoramidate linkage (indicated by the “z” subscript); the fourth nucleoside comprises a 2’-P-D-deoxyribosyl sugar moiety (indicated by the “d” subscript) and an unmodified guanine nucleobase linked to the fifth nucleoside with a phosphorothioate linkage; and the fifth nucleoside comprises a 2’-P-D-deoxyribosyl sugar moiety and an unmodified cytosine nucleobase; and the compound may include additional substituents, such as a conjugate group.

[0789] Herein, sugar, intemucleoside linkage, and nucleobase modifications may be indicated within a nucleotide or nucleobase sequence (e.g., by superscript or subscript, as shown above) or may be indicated in text accompanying a sequence (e.g., in separate text that appears within or above or below a table of compounds).

[0790] Where a specific compound is described herein by way of a drawn chemical structure, each nucleobase, sugar, and intemucleoside linkage of such a specific compound includes only the modifications indicated in the drawn chemical structure. One of skill will appreciate, however, that drawn compounds may exist in equilibrium between tautomeric forms and / or as salts in equilibrium with protonated or ionic forms. Drawn structures are intended to capture all such forms of such compounds.

[0791] While effort has been made to accurately describe compounds in the accompanying sequence listing, should there be any discrepancies between a description in this specification and in the accompanying sequence listing, the description in the specification and not in the sequence listing is the accurate description.

[0792] The compounds described herein include variations in which one or more atoms are replaced with a non-radioactive isotope or radioactive isotope of the indicated element. For example, compounds herein that comprise hydrogen atoms encompass all possible deuterium substitutions for each of the ’H hydrogen atoms. Isotopic substitutions encompassed by the compounds herein include but are not limited to:2H or3H in place of ’H,13C or14C in place of12C,15N in place of14N,17O or18O in place of16O, and33S,34S,35S, or36S in place of32S. In certain embodiments, non-radioactive isotopic substitutions may impart new properties on the oligomeric agent that are beneficial for use as a therapeutic or research tool. In certain CHEMO 113W0 embodiments, radioactive isotopic substitutions may make the compound suitable for research or diagnostic purposes such as imaging.

[0793] Examples

[0794] The following examples illustrate certain embodiments of the present disclosure and are not limiting. Moreover, where specific embodiments are provided, the inventors have contemplated generic application of those specific embodiments. For example, disclosure of an oligonucleotide having a particular motif provides reasonable support for additional oligonucleotides having the same or similar motif. And, for example, where a particular high-affinity modification appears at a particular position, other high-affinity modifications at the same position are considered suitable, unless otherwise indicated.

[0795] Example 1: Design of modified oligonucleotides complementary to human CNS Target A RNA

[0796] Modified oligonucleotides complementary to human Target A were designed as described in the tables below.

[0797] The modified oligonucleotides in the table below are mixed cEt / MOE gapmers with mixed intemucleoside linkages. The modified oligonucleotides in the table below are 20 nucleosides in length; have the sugar motif presented in the column labeled “Sugar Motif (5' to 3')”, wherein each “e” represents a 2'-M0E sugar moiety, each “k” represents a cEt sugar moiety, and each “d” represents a 2'-p-D- deoxyribosyl sugar moiety; and the intemucleoside linkage motif presented in the column labeled “Intemucleoside Linkages (5' to 3')”, wherein each “s” represents a phosphorothioate intemucleoside linkage, each “z” represents a mesyl phosphoramidate intemucleoside linkage, and each “o” represents a phosphodiester intemucleoside linkage. Each cytosine residue is a 5 -methylcytosine. “N” represents a nucleobase selected from adenine, guanine, 5 -methylcytosine, and thymine. Each modified oligonucleotide listed in the tables below is 100% complementary to Target A RNA. Each modified oligonucleotide with the same sequence code has an identical nucleobase sequence.

[0798] Table 1

[0799] Mixed cEt / MOE gapmers with mixed intemucleoside linkages complementary to human Target A

[0800] Example 2: Design of modified oligonucleotides complementary to human Target A RNA Modified oligonucleotides complementary to human Target A were designed as described in the tables below.

[0801] The modified oligonucleotides in the table below mixed cEt / MOE gapmers with mixed intemucleoside linkages. The modified oligonucleotides in the table below are 19-20 nucleosides in length, have the sugar motif presented in the column labeled “Sugar Motif (5' to 3')”, wherein each “e” represents a 2'-MOE sugar moiety, each “k” represents a cEt sugar moiety, and each “d” represents a 2'-P- D-deoxyribosyl sugar moiety; and the intemucleoside linkage motif presented in the column labeled “Intemucleoside Linkages (5' to 3')”, wherein each “s” represents a phosphorothioate intemucleoside linkage, each “z” represents a mesyl phosphoramidate intemucleoside linkage, and each “o” represents a phosphodiester intemucleoside linkage. Each cytosine residue is a 5 -methylcytosine. “N” represents a nucleobase selected from adenine, guanine, 5 -methylcytosine, and thymine.

[0802] Each modified oligonucleotide listed in the tables below is 100% complementary to Target A RNA.

[0803] Table 2

[0804] Mixed cEt / MOE gapmers with mixed intemucleoside linkages complementary to Target A, 20 nt

[0805] Example 3: Design of modified oligonucleotides complementary to CNS Target A

[0806] Modified oligonucleotides complementary to a Target A nucleic acid were designed as described in the tables below.

[0807] “N” represents a nucleobase selected from adenine, guanine, 5 -methylcytosine, and thymine. Each cytosine is a 5 -methylcytosine. Each sequence is 100% complementary to Target A RNA.

[0808] Each modified oligonucleotide in the tables below have the sugar motif designated in the column labeled “Sugar Motif (5' to 3')”, wherein each “e” represents a 2'-MOE sugar moiety, each “y” represents a 2'-OMe sugar moiety, each “k” represents a cEt sugar moiety, each “1” represents an LNA sugar moiety, and each “d” represents a 2'-p-D-deoxyribosyl sugar moiety; and the intemucleoside linkage motif in the column labeled “Intemucleoside Linkages (5' to 3')”, wherein each “s” represents a phosphorothioate intemucleoside linkage, each “z” represents a mesyl phosphoramidate intemucleoside linkage, and each “o” represents a phosphodiester intemucleoside linkage. Compound AO 14 was previously disclosed in International Patent Application Nos. PCT / US24 / 016095 and PCT / US24 / 016097. Compound B027 was previously disclosed in International Patent Application No. PCT / US24 / 016097. CHEMO 113W0

[0809] Table 3

[0810] 5-10-5 gapmers with mixed intemucleoside linkages complementary to Target A

[0811] Table 4

[0812] 5-10-5 gapmers with mixed intemucleoside linkages complementary to Target A

[0813] Example 4: Deamination analysis of modified oligonucleotides

[0814] Modified oligonucleotides described above were evaluated in an adenosine deaminase enzyme assay to determine the half-life of 5 '-terminal adenosine deamination.

[0815] Modified oligonucleotides were incubated at 37 °C with recombinant human ADA2 enzyme (rhCECR) (R&D Systems, catalog #7518-AD) in 10 mM HEPES, pH 7 at a final concentration of 10 pM of modified oligonucleotide and 0.1 mg / mL of ADA2 enzyme. After each 15 to 30-minute timepoint, 10 pL of the reaction mixture was withdrawn and stopped by addition of 40 pL of 8 M urea. Samples were analyzed by HPLC-MS / MS using an Exacta Orbitrap mass spectrometer (Thermo Fisher Scientific), a Vanquish LC system with a Waters ACQUITY Premier Oligonucleotide C18, 130A, 1.7mm 2.1x100 mm column (Waters), and Xcalibur software (Thermo Fisher Scientific). For each modified oligonucleotide, the amount of deaminated compound was quantified by integrating the peak corresponding to its predicted m / z values, wherein the deaminated compound has a 5 '-terminal inosine nucleobase in place of the 5 '-terminal adenosine. The amount of deaminated compound was then normalized to the total amount of compound. Deamination T1 / 2 values were calculated in GraphPad Prism (GraphPad, San Diego) using an equation for first-order kinetics, and reported in the table below. Where T1 / 2 could not be calculated due to low amounts of deamination, the level of deamination measured after 48 hours was reported in the table below as % of total. “N.D” indicates that data was not determined. CHEMO 113W0

[0816] Table 4

[0817] Deamination T1 / 2 values for modified oligonucleotides

[0818] Table 5

[0819] Deamination TI / 2values for modified oligonucleotides

[0820] In certain embodiments, an oligomeric agent comprising a bicyclic adenosine, such as a cEt adenosine, at its 5 ’-most position as described herein has greater metabolic stability when compared to an analogous oligomeric agent that is identical other than the sugar moiety of the 5 ’-most adenosine nucleoside. In certain embodiments, the analogous oligomeric agent is identical other than comprising a 2’- substituted sugar moiety or a deoxy sugar moiety at the 5 ’-most position.

[0821] Example 5: Deamination analysis of modified oligonucleotides

[0822] Modified oligonucleotides described above and B028 were evaluated in an adenosine deaminase enzyme assay to determine the half-life of 5 '-terminal adenosine deamination. B028 is a similar oligonucleotide to B027 having a different intemucleoside linkage motif. B028 has a mesyl phosphoramidate as the 5 ’-most intemucleoside linkage.

[0823] Modified oligonucleotides were incubated at 37 °C with recombinant human ADA2 enzyme (rhCECR) (R&D Systems, catalog #7518-AD) in 10 mM HEPES, pH 7 at a final concentration of 10 pM of modified oligonucleotide and 0.1 mg / mL of ADA2 enzyme. After each 15 to 30-minute timepoint, 10 pL of the reaction mixture was withdrawn and stopped by addition of 40 pL of 8 M urea. Samples were analyzed by HPLC-MS / MS using an Exacta Orbitrap mass spectrometer (Thermo Fisher Scientific), a Vanquish LC system with a Waters ACQUITY Premier Oligonucleotide C18, 130A, 1.7mm 2.1x100 mm column (Waters), and Xcalibur software (Thermo Fisher Scientific). For each modified oligonucleotide, the amount of deaminated compound was quantified by integrating the peak corresponding to its predicted m / z values, wherein the deaminated compound has a 5 '-terminal inosine nucleobase in place of the 5 '-terminal adenosine. The amount of deaminated compound was then normalized to the total amount of compound.

[0824] Table 6

[0825] Deamination T1 / 2 values for modified oligonucleotide

Claims

1. CLAIMS1. An oligomeric agent for use in treatment of a disease or condition comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):Ab-Wl-G-W2I wherein Ab is a bicyclic adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside ofWl is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 3 ’-most nucleoside ofWl is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;G consists of 5-20 linked 2’-deoxynucleosides;W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; wherein the oligomeric agent has improved metabolic stability compared to an analogous oligomeric agent having a non-bicyclic nucleoside 5’ ofWl; optionally wherein the modified oligonucleotide consists of the region of formula I.

2. The oligomeric agent of claim 1, wherein at least one of W1 and / or W2 comprises a 2’- deoxynucleoside and at least one intemucleoside linkage is not a phosphorothioate or phosphodiester.

3. The oligomeric agent of claim 2, wherein the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to an Lpa transcript (SEQ ID NO: 1).

4. The oligomeric agent of claim 1, wherein at least one of W1 and / or W2 comprises a 2’-M0E nucleoside and at least one intemucleoside linkage is a phosphodiester.

5. The oligomeric agent of claim 1, wherein at least one of W1 and / or W2 comprises a 2’-M0E nucleoside and the modified oligonucleotide comprises an intemucleoside linkage other than a phosphorothioate, phosphodiester, or a methoxypropyl phosphonate linkage.

6. The oligomeric agent of claim 1, wherein at least one of W1 and / or W2 comprises a 2’-M0E nucleoside and the modified oligonucleotide comprises a mesyl phosphoramidate intemucleoside linkage.

7. The oligomeric agent of any of claim 4-6, wherein the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to an APOE transcript (SEQ ID NO: 2).

8. The oligomeric agent of claim 1, wherein the 5 ’-most intemucleoside linkage is a mesyl phosphoramidate intemucleoside linkage, and wherein the modified oligonucleotide comprises at least one modified nucleoside comprising a modified furanosyl sugar moiety other than a cEt sugar moiety or at least one sugar surrogate nucleoside.

9. The oligomeric agent of claim 1, wherein the 5 ’-most intemucleoside linkage is a mesyl phosphoramidate intemucleoside linkage, and wherein the modified oligonucleotide comprises no more than 3 cEt nucleosides.

10. The oligomeric agent of claim 1, wherein the 5 ’-most intemucleoside linkage is a mesyl phosphoramidate intemucleoside linkage;W1 is not two consecutive cEt nucleosides, and wherein the modified oligonucleotide comprises no more than 9 mesyl phosphoramidate intemucleoside linkage.

11. The oligomeric agent of claim 1, wherein Ab comprises a cEt sugar moiety, and G is from 11-20.

12. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):Ab-Wl-G-W2I wherein Ab is a bicyclic adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside ofWl is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 3 ’-most nucleoside ofWl is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;G consists of 5-20 linked 2’-deoxynucleosides;W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogatenucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; provided that the modified oligonucleotide comprises at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate; and provided that the modified oligonucleotide does not comprise a methylphosphonate or methoxypropyl phosphonate intemucleoside linkage; optionally wherein the modified oligonucleotide consists of the region of formula I.

13. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):Ab-Wl-G-W2I wherein Ab is a cEt adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside ofWl is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 3 ’-most nucleoside ofWl is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside, and provided that at least one nucleoside of W1 is not a cEt;G consists of 5-20 linked 2’-deoxynucleosides;W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; wherein the modified oligonucleotide comprises at least one phosphodiester intemucleoside linkage; optionally wherein the modified oligonucleotide consists of the region of formula I.

14. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):Ab-Wl-G-W2I wherein Ab is a cEt adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside ofWl is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the3 ’-most nucleoside ofWl is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;G consists of 11-20 linked 2’-deoxynucleosides;W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; optionally wherein the modified oligonucleotide consists of the region of formula I.

15. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):Ab-Wl-G-W2I wherein Ab is a cEt adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside ofWl is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 3 ’-most nucleoside ofWl is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside, and provided that W1 does not comprise a cEt nucleoside;G consists of 5-20 linked 2’-deoxynucleosides;W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; provided that the modified oligonucleotide comprises at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate; optionally wherein the modified oligonucleotide consists of the region of formula I.

16. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):Ab-Wl-G-W2I wherein Ab is a bicyclic adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside ofWl is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the3 ’-most nucleoside ofWl is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;G consists of 5-20 linked 2’-deoxynucleosides;W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; provided that the modified oligonucleotide comprises at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate; and provided that the modified oligonucleotide comprises at least one but no more than 3 2’- MOE nucleosides and W1 is not 2 consecutive cEt nucleosides; optionally wherein the modified oligonucleotide consists of the region of formula I.

17. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):Ab-Wl-G-W2I wherein Ab is a cEt adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside ofWl is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 3 ’-most nucleoside ofWl is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;G consists of 5-20 linked 2’-deoxynucleosides;W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside, and provided that W2 comprises at least two non-cEt nucleosides; provided that the modified oligonucleotide comprises at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate; and the modified oligonucleotide does not comprise a methoxy propyl phosphonate intemucleoside linkage; optionally wherein the modified oligonucleotide consists of the region of formula I.

18. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):Ab-Wl-G-W2I wherein Ab is a bicyclic adenosine; wherein W1 consists of 1-7 linked nucleosides, wherein each nucleoside ofWl is independently selected from a modified nucleoside comprising a modified fiiranosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 3 ’-most nucleoside ofWl is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside;G consists of 5-20 linked 2’-deoxynucleosides;W2 consists of 2-8 linked nucleosides, wherein each nucleoside ofW2 is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside, provided that the 5 ’-most nucleoside of W2 is a modified nucleoside comprising a modified fiiranosyl sugar moiety or a sugar surrogate nucleoside; provided that the modified oligonucleotide comprises at least one phosphodiester intemucleoside linkage; and further provided that W1 or W2 comprises at least one 2-deoxynucleoside; optionally wherein the modified oligonucleotide consists of the region of formula I.

19. The oligomeric agent of any of claims 12-18, wherein the oligomeric agent has improved metabolic stability compared to an analogous oligomeric agent having a non-bicyclic nucleoside 5’ ofWl.

20. The oligomeric agent of any of claims 1-19, wherein the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to an Lpa transcript (SEQ ID NO: 1).

21. The oligomeric agent of any of claims 1-20, wherein the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to an APOE transcript (SEQ ID NO: 2).

22. The oligomeric agent of any of claims 1-20, wherein the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to a PLN transcript (SEQ ID NO: 3).

23. The oligomeric agent of any of claims 1-10, 12, or 16 wherein Ab is a cEt adenosine.

24. The oligomeric agent of any of claims 1-10, 12, or 16 wherein Ab is an LNA adenosine.

25. The oligomeric agent of any of claims 1-14 or 16-24, wherein each nucleoside of W1 is independently selected from a bicyclic nucleoside other than a cEt nucleoside, a non-bicyclic 2’- substituted nucleoside, a sugar surrogate nucleoside, and a 2 ’-deoxynucleoside.

26. The oligomeric agent of any of claims 1-25, comprising at least one phosphorothioate intemucleoside linkage.

27. The oligomeric agent of any of claims 1-12, 14-17, or 19-26, comprising at least one phosphodiester intemucleoside linkage.

28. The oligomeric agent of any of any of claims 1-27 , comprising at least one intemucleoside linkage that is not a phosphodiester or a phosphorothioate intemucleoside linkage.

29. The oligomeric agent of any of claims 1-28, wherein the modified oligonucleotide does not comprise a methyl propyl phosphonate or a methylphosphonate intemucleoside linkage.

30. The oligomeric agent of any of claims 1-29, wherein the modified oligonucleotide comprises at least one mesyl phosphoramidate intemucleoside linkage.

31. The oligomeric agent of any of claims 1-30, wherein each intemucleoside linkage is independently selected from a mesyl phosphoramidate, a phosphorothioate, and a phosphodiester intemucleoside linkage.

32. The oligomeric agent of any of claims 1-31, wherein the 3’-most nucleoside of W1 does not comprise a 2’-OMe nucleoside.

33. The oligomeric agent of any claims 1-32, wherein the modified oligonucleotide comprises no more than three cEt nucleosides.

34. The oligomeric agent of any of claims 1-33, wherein the modified oligonucleotide comprises no more than three 2 ’-MOE nucleosides.

35. The oligomeric agent of any of claims 1-15 or 17-34, wherein the modified oligonucleotide comprises at least four 2’-M0E nucleosides.

36. The oligomeric agent of any of claims 1-35, , wherein W1 comprises no more than three 2 ’-MOE nucleosides.

37. The oligomeric agent of any of claims 1-36, wherein W1 does not comprise three contiguous cEt nucleosides.

38. The oligomeric agent of any of claims 1-37, wherein W1 does not comprise two contiguous cEt nucleosides.

39. The oligomeric agent of any of claims 1-38, wherein W1 does not comprise three contiguous 2’- MOE nucleosides.

40. The oligomeric agent of any of claims 1-39, wherein each intemucleoside linkage at the 3’- position of each nucleoside of G is independently selected from a mesyl phosphoramidate intemucleoside linkage and a phosphorothioate intemucleoside linkage.

41. The oligomeric agent of any of claims 1-40, wherein each nucleoside of W1 is independently selected from a non-bicyclic 2 ’-substituted nucleoside, a bicyclic nucleoside, and a 2’- deoxynucleoside.

42. The oligomeric agent of any of claims 1-40, wherein each nucleoside of W1 is independently selected from a non-bicyclic 2 ’-substituted nucleoside, a bicyclic nucleoside other than a cEt nucleoside, and a 2’ -deoxynucleoside.

43. The oligomeric agent of any of claims 1-40, wherein each nucleoside of W1 is independently selected from a non-bicyclic 2 ’-substituted nucleoside and a 2 ’-deoxynucleoside.

44. The oligomeric agent of any of claims 1-40, wherein each nucleoside of W1 is independently a non-bicyclic 2 ’-substituted nucleoside or a bicyclic nucleoside other than a cEt nucleoside.

45. The oligomeric agent of any of claims 1-44, wherein the 3 ’-most nucleoside of W1 is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage.

46. The oligomeric agent of any of claims 1-45, wherein G consists of at least 11 linked nucleosides, optionally 11, 12, 13, 14, or 15 linked nucleosides.

47. The oligomeric agent of claims 1-10, 12-13, or 15-45, wherein G consists of 10 linked nucleosides.

48. The oligomeric agent of any of claims 1-47, wherein the modified oligonucleotide comprises exactly one, exactly two, or exactly three cEt nucleosides.

49. The oligomeric agent of any of claims 1-48, wherein at least one ofWl and W2 comprises a 2’- deoxynucleoside.

50. The oligomeric agent of any of claims 1-49, wherein each 2 ’-deoxynucleoside of W1 and W2 is linked at its respective 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage.

51. The oligomeric agent of any of claims 1-50, wherein each 2 ’-deoxynucleoside ofWl is linked at its 3’-position by a mesyl phosphoramidate intemucleoside linkage.

52. The oligomeric agent of any of claims 1-51, wherein each 2 ’-deoxynucleoside ofW2 is linked at its 3’-position by a mesyl phosphoramidate intemucleoside linkage.

53. The oligomeric agent of any of claims 1-52, wherein W1 comprises 1-4 modified nucleosides comprising a modified furanosyl sugar moiety.

54. The oligomeric agent of any of claims 1-52, wherein W1 comprises 2 or 3 modified nucleosides comprising a modified furanosyl sugar moiety, optionally 3.

55. The oligomeric agent of any of claims 1-52, wherein W1 comprises 3 contiguous modified nucleosides comprising a modified furanosyl sugar moiety.

56. The oligomeric agent of any of claims 1-55, wherein each modified nucleoside ofWl is linked at its 3 ’-position by a phosphodiester or a phosphorothioate intemucleoside linkage, optionally a phosphorothioate intemucleoside linkage.

57. The oligomeric agent of any of claims 1-55, wherein the 3 ’-most nucleoside of W1 is linked at its 3 ’-position by a phosphorothioate intemucleoside linkage.

58. The oligomeric agent of any of claims 1-57, wherein W 1 consists of 2, 3, or 4 linked nucleosides.

59. The oligomeric agent of any of claims 1-57, wherein W1 does not comprise a 2’- deoxynucleoside.

60. The oligomeric agent of any of claims 1-58, wherein W1 comprises at least one 2’- deoxynucleoside.

61. The oligomeric agent of any of claims 1-60, wherein each 2 ’-substituted nucleoside of W1 is a 2’- MOE nucleoside.

62. The oligomeric agent of any of claims 1-61, wherein W1 comprises at least four 2 ’-MOE nucleosides.

63. The oligomeric agent of any of claims 1-62, wherein each nucleoside of W1 is a 2 ’-MOE nucleoside.

64. The oligomeric agent of any of claims 1-63, wherein W1 consists of four contiguous 2 ’-MOE nucleosides.

65. The oligomeric agent of any of claims 1-64, wherein each 2 ’-deoxynucleoside of W1 is a 2’-P-D- deoxyribosyl nucleoside.

66. The oligomeric agent of any of claims 1-65, wherein each bicyclic nucleoside of W1 is an LNA nucleoside.

67. The oligomeric agent of any of claims 1-66, wherein W2 comprises 1-5 modified nucleosides comprising a modified furanosyl sugar moiety.

68. The oligomeric agent of any of claims 1-67, wherein W2 comprises 3 modified nucleosides comprising a modified furanosyl sugar moiety.

69. The oligomeric agent of any of claims 1-67, wherein W2 comprises 5 modified nucleosides comprising a modified furanosyl sugar moiety.

70. The oligomeric agent of any of claims 1-68, wherein W2 consists of 3, 4, 5, or 6 linked nucleosides.

71. The oligomeric agent of any of claims 1-68, wherein W2 consists of 3, 4, or 5 linked nucleosides.

72. The oligomeric agent of any of claims 1-71, wherein each non-terminal modified nucleoside of W2 is linked at its 3 ’-position by a phosphodiester or a phosphorothioate intemucleoside linkage, optionally a phosphorothioate intemucleoside linkage.

73. The oligomeric agent of any of claims 1-72, wherein W2 does not comprise a 2’- deoxynucleoside.

74. The oligomeric agent of any of claims 1-72, wherein W2 comprises at least one 2’- deoxynucleoside.

75. The oligomeric agent of any of claims 1-74, wherein each modified nucleoside of W2 is independently selected from a non-bicyclic 2 ’-substituted nucleoside and a bicyclic nucleoside.

76. The oligomeric agent of any of claims 1-74, wherein each modified nucleoside of W2 is independently selected from a non-bicyclic 2 ’-substituted nucleoside and a bicyclic nucleoside other than a cEt nucleoside.

77. The oligomeric agent of any of claims 1-76, wherein each modified nucleoside of W2 is independently a bicyclic nucleoside.

78. The oligomeric agent of any of claims 1-77, wherein each bicyclic nucleoside of W2 is selected from cEt and LNA.

79. The oligomeric agent of any of claims 1-74, wherein each modified nucleoside of W2 is independently a non-bicyclic 2’ -substituted nucleoside.

80. The oligomeric agent of any of claims 1-79, wherein each 2 ’-substituted nucleoside of W2 is a 2’- MOE nucleoside.

81. The oligomeric agent of any of claims 1-79, wherein each 2 ’-deoxynucleoside of W2 is a 2’-P-D- deoxyribosyl nucleoside.

82. The oligomeric agent of any of claims 1-81, wherein each nucleoside of G is a 2’-P-D- deoxyribosyl nucleoside.

83. The oligomeric agent of any of claims 1-81, wherein each nucleoside of G is linked at its 3’- position by a phosphorothioate or a mesyl phosphoramidate intemucleoside linkage.

84. The oligomeric agent of any of claims 1-83, wherein at least one, optionally exactly one, nucleoside of G is linked at its 3 ’-position by a mesyl phosphoramidate intemucleoside linkage.

85. The oligomeric agent of any of claims 1-84, wherein exactly two, exactly three, exactly four, exactly five, or exactly six nucleosides of G are linked at their respective 3 ’-positions by a mesyl phosphoramidate intemucleoside linkage.

86. The oligomeric agent of any of claims 1-84, wherein exactly five, exactly six, exactly seven, exactly eight, exactly nine, or exactly ten nucleosides of G are linked at their respective 3’- positions by a phosphorothioate intemucleoside linkage.

87. The oligomeric agent of any of claims 1-84, wherein the 3’-most nucleoside of G is linked at its 3 ’-position by a mesyl phosphoramidate or phosphorothioate intemucleoside linkage.

88. The oligomeric agent of any of claims 1-13, 15-45, or 48-87, wherein G consists of 7-15 linked nucleosides, optionally 10-13 linked nucleosides, optionally exactly 10 linked nucleosides.

89. The oligomeric agent of any of claims 1-88, wherein G is a deoxy region.

90. The oligomeric agent of any of claims 1-89, wherein W2 has the formula -(E)n4-(D4)n5-(F)n6 wherein each F is independently selected from a modified nucleoside comprising a modified fiiranosyl sugar moiety, a sugar surrogate nucleoside, and a 2’-deoxy nucleoside, provided that at least one F is a modified nucleoside comprising a modified fiiranosyl sugar moiety or a sugar surrogate nucleoside; and each non-terminal 2’-deoxy nucleoside of F is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; each E is independently selected from a modified nucleoside comprising a modified fiiranosyl sugar moiety and a sugar surrogate nucleoside; each D4 is independently a 2 ’-deoxy nucleoside provided that each D4 is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; n6 is 1 to 8 and each of n4 and n5 is independently 0 to 8, provided that n4 + n5 + n6 is less than or equal to 891. The oligomeric agent of claim 90, wherein n4 is 1 or 2.

92. The oligomeric agent of claim 90 or 91, wherein n5 is 1.

93. The oligomeric agent of claim 90 or 91, wherein n5 is 0.

94. The oligomeric agent of any one of claims 90-93, wherein n6 is 1 to 5.

95. The oligomeric agent of any one of claims 90-94, wherein n6 is 1, 2, or 3.

96. The oligomeric agent of any one of claims 90-95, wherein each E is selected from a bicyclic nucleoside and a 2 ’-substituted nucleoside.

97. The oligomeric agent of any one of claims 90-96, wherein each E is a bicyclic nucleoside.

98. The oligomeric agent of any one of claims 90-97, wherein each bicyclic nucleoside of E is selected from cEt and LNA, optionally cEt.

99. The oligomeric agent of any one of claims 90-96, wherein each E is a non-bicyclic 2 ’-substituted nucleoside.

100. The oligomeric agent of any one of claims 90-99, wherein each 2 ’-substituted nucleoside of E is a 2 ’-MOE nucleoside.

101. The oligomeric agent of any one of claims 90-100, wherein each D4 is a 2’-P-D- deoxyribosyl nucleoside.

102. The oligomeric agent of any one of claims 90-101, wherein each F is selected from a bicyclic nucleoside and a non-bicyclic 2 ’-substituted nucleoside.

103. The oligomeric agent of any one of claims 90-102, wherein each F is a non-bicyclic 2’- substituted nucleoside.

104. The oligomeric agent of any one of claims 90-102, wherein non-bicyclic 2 ’-substituted nucleoside of F is a 2 ’-MOE nucleoside.

105. The oligomeric agent of any one of claims 90-102 or 104, wherein each F is a bicyclic nucleoside.

106. The oligomeric agent of any one of claims 90-105, wherein each bicyclic nucleoside of F is selected from cEt and LNA, optionally cEt.

107. The oligomeric agent of any one of claims 90-106, wherein each of n4 and n5 is 0.

108. The oligomeric agent of any one of claims 90-107, wherein n6 is 5, and each of n4 and n5 is 0.

109. The oligomeric agent of any of claims 1-108, wherein W1 has the formula (A)nl-(Dl)n2- (B)n3- wherein each A is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety, a sugar surrogate nucleoside, and a 2’ -deoxy nucleoside provided that at least one A is a modified nucleoside comprising a modified furanosyl sugar moiety or a sugar surrogate nucleoside; and each 2’-deoxy nucleoside of A is linked at its 3’- position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; each B is independently selected from a modified nucleoside comprising a modified furanosyl sugar moiety and a sugar surrogate nucleoside; each DI is independently a 2 ’-deoxy nucleosideprovided that each DI is linked at its 3 ’-position by a phosphorothioate or mesyl phosphoramidate intemucleoside linkage; nl is 1 to 8 and each of n2 and n3 is independently 0 to 8, provided that nl + n2 + n3 is less than or equal to 8110. The oligomeric agent of claim 109, wherein n3 is 0 or 1.

111. The oligomeric agent of claim 109 or 110, wherein n2 is 0 or 1.

112. The oligomeric agent of any one of claims 109-111, wherein nl is 1 to 5.

113. The oligomeric agent of any one of claims 109-111, wherein nl is 2 or 3.

114. The oligomeric agent of any one of claims 109-113, wherein each B is selected from a bicyclic nucleoside and a non-bicyclic 2 ’-substituted nucleoside.

115. The oligomeric agent of any one of claims 109-114, wherein each B is a non-bicyclic 2’- substituted nucleoside.

116. The oligomeric agent of any one of claims 109-115, wherein each non-bicyclic 2’- substituted nucleoside of B is a 2 ’-MOE nucleoside.

117. The oligomeric agent of any one of claims 109-116, wherein each B is a bicyclic nucleoside.

118. The oligomeric agent of any one of claims 109-117, wherein each bicyclic nucleoside of B is selected from cEt and LNA, optionally cEt.

119. The oligomeric agent of any one of claims 109-118, wherein each D4 is a 2’-P-D- deoxyribosyl nucleoside.

120. The oligomeric agent of any one of claims 109-119, wherein each A is selected from a bicyclic nucleoside and a 2 ’-substituted nucleoside.

121. The oligomeric agent of any one of claims 109-120, wherein each A is a 2 ’-substituted nucleoside.

122. The oligomeric agent of any one of claims 109-121, wherein each 2 ’-substituted nucleoside of A is a 2’-M0E nucleoside.

123. The oligomeric agent of any one of claims 109-120 or 122, wherein each A is a bicyclic nucleoside.

124. The oligomeric agent of any one of claims 109-123, wherein each bicyclic nucleoside of A is selected from cEt and LNA, optionally cEt.

125. The oligomeric agent of any one of claims 109-124, wherein each of n2 and n3 is 0.

126. The oligomeric agent of any one of claims 109-125, wherein nl is 4, and each of n2 and n3 is 0.

127. The oligomeric agent of any of claims 1-126, wherein the modified oligonucleotide consists of 17-20 linked nucleosides.

128. The oligomeric agent of any of claims 1-127, wherein the modified oligonucleotide consists of 17, 18, 19, or 20 linked nucleosides.

129. The oligomeric agent of any of claims 1-126, wherein the modified oligonucleotide consists of 20 linked nucleosides.

130. The oligomeric agent of any one of claims 1-46 or 48-129, wherein G consists of 11-15 linked nucleosides, optionally 11, optionally 12, optionally 13, optionally 14, or optionally 15 linked nucleosides, optionally wherein neither W1 nor W2 comprises a 2 ’-deoxynucleoside.

131. An oligomeric agent comprising a modified oligonucleotide consisting of 14 to 30 linked nucleosides, wherein the modified oligonucleotide comprises a region of formula I (5’ to 3’):Ak-Wl-G-W2I wherein Akis a cEt adenosine, W1 consists of 4 linked nucleosides, wherein each nucleoside of W1 is selected from a non-bicyclic 2 ’-substituted nucleoside and a bicyclic nucleoside;G consists of 10 linked 2'-P-D-deoxyribosyl nucleosides;W2 consists of 5 linked nucleosides, wherein each nucleoside of W2 is independently selected from a non-bicyclic 2 ’-substituted nucleoside and a bicyclic nucleoside; provided that the modified oligonucleotide comprises at least one intemucleoside linkage other than a phosphorothioate or phosphodiester intemucleoside linkage; optionally wherein the modified oligonucleotide consists of the region of formula I132. The oligomeric agent of claim 131, wherein the modified oligonucleotide comprises at least one phosphodiester intemucleoside linkage.

133. The oligomeric agent of claim 131, wherein the modified oligonucleotide does not comprise a sequence of at least 16 nucleosides having 100% complementarity to an Lpa transcript (SEQ ID NO: 1).

134. The oligomeric agent of any of claims 131-133, wherein W1 comprises exactly one bicyclic nucleoside.

135. The oligomeric agent of any of claims 131-133, wherein each nucleoside of W1 is a non- bicyclic 2 ’-substituted nucleoside.

136. The oligomeric agent of any one of claims 131-135, wherein each non-bicyclic 2’- substituted nucleoside of W1 is selected from a 2 ’-MOE nucleoside and a 2’-0Me nucleoside.

137. The oligomeric agent of claim 136, wherein each non-bicyclic 2 ’-substituted nucleoside of W1 is a 2 ’-MOE nucleoside.

138. The oligomeric agent of any one of claims 131-137, wherein each bicyclic nucleoside of W1 is selected from a cEt nucleoside and an LNA nucleoside.

139. The oligomeric agent of any one of claims 131-137, wherein each bicyclic nucleoside of W1 is a cEt nucleoside.

140. The oligomeric agent of any one of claims 131-139, wherein W2 comprises no bicyclic nucleosides.

141. The oligomeric agent of any one of claims 131-140, wherein W2 comprises exactly one bicyclic nucleoside.

142. The oligomeric agent of any one of claims 131-139, wherein W2 comprises exactly two bicyclic nucleosides.

143. The oligomeric agent of any one of claims 131-143, wherein each non-bicyclic 2’- substituted nucleoside of W2 is selected from a 2 ’-MOE nucleoside and a 2’-OMe nucleoside.

144. The oligomeric agent of claim 143, wherein each non-bicyclic 2 ’-substituted nucleoside of W2 is a 2 ’-MOE nucleoside.

145. The oligomeric agent of any one of claims 131-144, wherein each intemucleoside linkage is selected from a phosphodiester, a phosphorothioate, and a mesyl phosphoramidate.

146. The oligomeric agent of any of claims 1-145, wherein the modified oligonucleotide comprises nucleobases independently selected from thymine, uracil, guanine, cytosine, 5- methylcytosine, adenine, and hypoxanthine.

147. The oligomeric agent of any of claims 1-146, comprising a conjugate group.

148. The oligomeric agent of claim 147, wherein the conjugate group comprises a carbohydrate or carbohydrate cluster.

149. The oligomeric agent of claim 147 or 148, wherein the conjugate group comprises at least one GalNAc.

150. The oligomeric agent of claim 147, wherein the conjugate group comprises a C10-C20 alkyl chain.

151. The oligomeric agent of claim 150, wherein the conjugate group comprises Cie alkyl.

152. The oligomeric agent of claim 147, wherein the conjugate group comprises a transferrin receptor ligand.

153. The oligomeric agent of claim 152, wherein the transferrin receptor ligand comprises a peptide.

154. The oligomeric agent of claim 152 or 153, wherein the transferrin receptor ligand comprises a bicyclic peptide.

155. The oligomeric agent of claim 152, wherein the transferrin receptor ligand comprises a Fab, Fab’, F(ab’)2, scFv, VHH, VNAR, or nanobody.

156. The oligomeric agent of any one of claims 147-155, wherein the oligomeric agent comprises the modified oligonucleotide, the conjugate group, and a cleavable linker comprising nucleosides linked by phosphodiester linkages linking the modified oligonucleotide and the conjugate group.

157. The oligomeric agent of any one of claims 1-146, wherein the oligomeric agent consists of the modified oligonucleotide.

158. The oligomeric agent of any of claims 1-157, wherein the nucleobase sequence of the modified oligonucleotide is complementary to a target nucleic acid.

159. The oligomeric agent of claim 158, wherein the nucleobase sequence of the modified oligonucleotide is at least 80%, at least 90%, or 100% complementary to the target nucleic acid.

160. The oligomeric agent of claim 158 or 159, wherein the target nucleic acid is selected from: an mRNA, a pre-mRNA, a microRNA, and a non-coding RNA.

161. The oligomeric agent of claim 158, wherein the target nucleic acid is not a CNS target.

162. The oligomeric agent of claim 158, wherein the target nucleic acid is a CNS target.

163. The oligomeric agent of any of claims 1-162, wherein the modified oligonucleotide is a first modified oligonucleotide, and further comprising a second modified oligonucleotide that is complementary to the first modified oligonucleotide.

164. A pharmaceutical composition comprising the oligomeric agent of any of claims 1-163 and a pharmaceutically acceptable carrier.

165. A method comprising contacting a cell with the oligomeric agent or pharmaceutical composition of any of claims 1-163.

166. A method of modulating the amount or activity of a target nucleic acid in a cell, comprising contacting the cell with the oligomeric agent of any of claims 1-163 or pharmaceutical composition of claim 164.

167. The method of claim 164-165, whereby the amount or activity of a target nucleic acid is reduced.

168. The method of any one of claims 164-167, comprising parenteral administration to a subject.

169. The method of any one of claims 164-168, comprising injection into CSF.

170. The method of any one of claims 164-168, comprising subcutaneous injection.

171. The oligomeric agent of any of claims 1-163 or the pharmaceutical composition of claim 162, for use in treatment of a disease or condition.

172. The oligomeric agent of any of claims 1-163 or the pharmaceutical composition of claim 162, for use in the manufacture of a medicament.

173. The oligomeric agent of any of claims 1-163 or the pharmaceutical composition of claim 162, for use in therapy.

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