Oligonucleotide compositions and methods thereof

Abstract

Oligonucleotides, compositions and methods are provided.SOLUTION: Provided is an oligonucleotide comprising a pattern of backbone chiral centers (linking phosphorus) of (Op)n(Sp)m, wherein Sp indicates the S configuration of the chiral linking phosphorus of a chirally modified internucleotidic linkage, Op indicates the achiral linking phosphorus of a natural phosphate linkage, and each of n and m is independently 1-20, and wherein each wing independently comprises one or more nucleobases.SELECTED DRAWING: Figure 4

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 62 / 542,778, filed August 8, 2017, which is incorporated herein by reference in its entirety. [Background technology]

[0002] Oligonucleotides are useful in a variety of applications, including but not limited to, therapeutic, diagnostic and / or research applications, including the treatment of various conditions, disorders and / or diseases. Summary of the Invention

[0003] Among other things, the present disclosure encompasses the recognition that structural elements of oligonucleotides, such as base sequence, chemical modifications (e.g., sugar, base, and / or internucleotide linkage modifications and patterns thereof), and / or stereochemistry (e.g., backbone chiral center (chiral internucleotide linkage) stereochemistry and / or pattern thereof), can have a profound effect on the activity and properties of the oligonucleotide, such as stability, toxicity, delivery, etc. In some embodiments, the present disclosure demonstrates that oligonucleotides and compositions comprising oligonucleotides with controlled structural elements, such as controlled chemical modifications and / or controlled backbone stereochemical patterns, provide unexpected activities and properties, including, but not limited to, those described herein. In some embodiments, the present disclosure demonstrates that the combination of chemical modifications and stereochemistry can provide unexpected and significantly improved activities and properties. In some embodiments, the present disclosure provides oligonucleotides and compositions comprising oligonucleotides having particular base sequences and / or patterns of sugar modifications (e.g., 2'-OMe, 2'-F, 2'-MOE, etc.), and / or patterns or base modifications (e.g., 5-methylcytosine), and / or patterns of backbone modifications (e.g., natural phosphate linkages, modified internucleotide linkages, etc.), and / or patterns of backbone chiral centers (e.g., Rp or Sp and / or stereorandom and / or achiral backbone-bound phosphorus atoms).

[0004] In some embodiments, the present disclosure provides novel oligonucleotides and compositions thereof (e.g., chiral controlled oligonucleotide compositions), wherein the oligonucleotide comprises a wing-core-wing format, and the first and second wings differ chemically from the core and each other, for example, in sugar or sugar modification or combination or pattern thereof, backbone internucleotide linkage or combination or pattern thereof, and / or backbone internucleotide linkage stereochemistry. In particular, in some embodiments, the present disclosure provides oligonucleotides comprising a wing-core-wing structure, wherein each wing independently comprises one or more sugar modifications, and the sugar modification pattern of one wing differs from that of the other wing. In some embodiments, one wing comprises a sugar modification that is not present in the other wing. In some embodiments, each sugar moiety of the core independently does not comprise a substituent at the 2'-position (i.e., comprises two -H at the 2'-position). In some embodiments, each sugar moiety of the core independently is a natural DNA sugar moiety (D-2-deoxyribose moiety) optionally substituted at the 5'-position. In some embodiments, each sugar moiety of the core is independently a natural DNA sugar moiety (D-2-deoxyribose moiety). In some embodiments, such oligonucleotides contain one or more chiral internucleotide linkages (e.g., phosphorothioate linkages [anionic at a particular pH -OP(O)(S - )-O-, such as -OP(O)(SH)-O-], a neutral internucleotide linkage as described herein. In some embodiments, such oligonucleotides contain one or more (e.g., at least 5, 6, 7, 8, 9, or 10) chiral neutral internucleotide linkages. In some embodiments, the present disclosure provides chiral controlled oligonucleotide compositions of such oligonucleotides. In some embodiments, the provided oligonucleotides contain a backbone chiral center (linked phosphorus) pattern of (Np)t[(Op / Rp)n(Sp)m]y, where each variable is independently as described in this disclosure. Unless otherwise specified, the stereochemical pattern is in the 5' to 3' direction, as will be understood by one of skill in the art.

[0005] In some embodiments, the present disclosure demonstrates that oligonucleotides and chiral controlled oligonucleotide compositions thereof comprising a particular stereochemical pattern, such as (Np)t[(Op / Rp)n(Sp)m]y, can provide highly improved activity (e.g., when used to cleave a target nucleic acid), specificity (e.g., when used to cleave a target nucleic acid in which nucleic acids of similar sequence are present [e.g., transcripts of wild-type and mutant alleles, transcripts from alleles containing single nucleotide polymorphisms (SNPs), etc.]), and / or other properties (e.g., stability, delivery, etc.) compared to a suitable control oligonucleotide and / or composition thereof (e.g., an unmodified oligonucleotide of the same base sequence; optionally a stereorandom oligonucleotide of the same composition; optionally a chiral controlled oligonucleotide of the same composition; or a stereorandom and / or chiral controlled composition thereof). In some embodiments, the provided oligonucleotides comprise a stereochemical pattern of (Np)t[(Op / Rp)n(Sp)m]y, where each variable is independently as described in this disclosure. In some embodiments, each Np is Sp. In some embodiments, the pattern comprises at least one Op. In some embodiments, n is 1. In some embodiments, m is at least 2, 3, 4, or 5. In some embodiments, y is 1. In some embodiments, y is 2, 3, 4, or 5. In some embodiments, (Np)t[(Op / Rp)n(Sp)m]y is (Sp)t[(Op / Rp)(Sp)m]y. In some embodiments, (Np)t[(Op / Rp)n(Sp)m]y is (Sp)t[Op(Sp)m]y. In some embodiments, each Np, Rp, and / or Sp bonded phosphorus is independently a bonded phosphorus of a phosphorothioate bond. In some embodiments, each Op is independently a natural internucleotide bond (anionic at a particular pH -OP(O)(O -)-O-. In some embodiments, each sugar moiety linked to the linked phosphorus in the stereochemical pattern, e.g., (Np)t[(Op / Rp)n(Sp)m]y, at its 3'-position does not contain a 2'-modification. In some embodiments, each sugar moiety linked to the linked phosphorus in the stereochemical pattern, e.g., (Np)t[(Op / Rp)n(Sp)m]y, at its 5'-position does not contain a 2'-modification. In some embodiments, each sugar moiety linked to the attached phosphorus of the stereochemical pattern, e.g., (Np)t[(Op / Rp)n(Sp)m]y, is independently a natural DNA sugar moiety (D-2-deoxyribose moiety), wherein the 5'-position is optionally substituted. In some embodiments, the 5'-position is unsubstituted. In some embodiments, the 5'-position is substituted, e.g., the 5'-position is linked to the attached phosphorus of Op.

[0006] In some embodiments, as demonstrated in the present disclosure, an oligonucleotide comprising a stereochemical pattern comprising NpNpOp(Sp)m has ( The target sequence can be cleaved (at the internucleotide bond of the target corresponding to the internucleotide bond containing the underlined Np in TIFF2026016363000002.tif473).

[0007] In some embodiments, the present disclosure provides a method for controlled cleavage of a target nucleic acid, the method comprising providing an oligonucleotide or a chiral controlled oligonucleotide composition thereof, wherein the stereochemical pattern of the oligonucleotide comprises (Np)t[(Op / Rp)n(Sp)m]y as described in the present disclosure.

[0008] In some embodiments, the present disclosure provides a method for controlled cleavage of a nucleic acid target, the method comprising contacting the target with a provided oligonucleotide or composition thereof. In some embodiments, the present disclosure provides a method for selective cleavage of a nucleic acid target, the method comprising contacting the target with a provided oligonucleotide or composition thereof. In some embodiments, the present disclosure provides a method for allele-specific cleavage of a transcript of a specific allele, the method comprising contacting the target with a provided oligonucleotide or composition thereof. In some embodiments, the provided oligonucleotide has a pattern of backbone chiral centers comprising Op(Sp)m. In some embodiments, the composition is a chiral-controlled oligonucleotide composition of a plurality of oligonucleotides, the pattern of backbone chiral centers of the oligonucleotide comprising Op(Sp)m. In some embodiments, m is 2. In some embodiments, the provided oligonucleotide has a pattern of backbone chiral centers comprising (Np)t[(Op / Rp)n(Sp)m]y, where each variable is as described in this disclosure, n is 1, m is 2 or more, and t is 2 or more. In some embodiments, the composition is a chiral controlled oligonucleotide composition of a plurality of oligonucleotides, wherein the pattern of backbone chiral centers of the oligonucleotides comprises (Np)t[(Op / Rp)n(Sp)m]y, where each variable is as described in this disclosure, n is 1, m is 2 or greater, and t is 2 or greater. In some embodiments, Np is Sp.

[0009] In some embodiments, oligonucleotides comprising the asymmetric format and / or stereochemical patterns described in the present disclosure have the ability to reduce the level, expression and / or activity of a gene target or its gene product.

[0010] The oligonucleotides of the present disclosure can function through a variety of mechanisms. In some embodiments, provided oligonucleotides are capable of reducing the level, expression, and / or activity of a gene target or its gene product through a mechanism involving RNase H, which recognizes DNA / RNA duplexes. In some embodiments, the core of the oligonucleotide contains multiple deoxyribose (e.g., 2'-deoxyribose or 2'-DNA sugars as found in naturally occurring DNA) moieties and is capable of annealing to RNA (e.g., target mRNA) to form a substrate for RNase H, allowing RNase H to cleave the RNA.

[0011] In some embodiments, provided oligonucleotides are capable of reducing the level, expression, and / or activity of a gene target or its gene product through a mechanism involving steric hindrance, hi some embodiments, provided oligonucleotides prevent or reduce translation of the target mRNA.

[0012] The present disclosure relates to any oligonucleotide having an asymmetric format and operating through any mechanism and comprising any structure or format (or portion thereof) described herein, wherein the oligonucleotide comprises at least one non-naturally occurring modification of the base, sugar and / or internucleotide linkage.

[0013] In some embodiments, provided oligonucleotides comprise at least one stereorandom internucleotide linkage (chiral non-controlled internucleotide linkage) (e.g., a stereorandom phosphorothioate linkage, a stereorandom neutral internucleotide linkage, etc.). In some embodiments, provided oligonucleotides comprise at least one stereocontrolled internucleotide linkage (chiral controlled internucleotide linkage) (e.g., an Rp or Sp phosphorothioate linkage, an Rp or Sp neutral internucleotide linkage, etc.).

[0014] In some embodiments, the present disclosure encompasses the recognition that various optional additional chemical moieties, such as carbohydrate moieties, sugar moieties, targeting moieties, etc., can be incorporated into oligonucleotides to improve one or more properties. In some embodiments, the additional chemical moiety is selected from glucose, GluNAc (N-acetylamine glucosamine), and anisamide moieties. In some embodiments, an oligonucleotide can include two or more additional chemical moieties, where the additional chemical moieties are identical or different, or of the same or different categories (e.g., carbohydrate moieties, sugar moieties, targeting moieties, etc.). In some embodiments, a particular additional chemical moiety facilitates delivery of the oligonucleotide to a desired cell, tissue, and / or organ, facilitates internalization of the oligonucleotide, and / or increases oligonucleotide stability.

[0015] In some embodiments, the present disclosure demonstrates that surprisingly high target specificity can be achieved with oligonucleotides having an asymmetric format. In some embodiments, the oligonucleotides having an asymmetric format are allele-specific, e.g., the oligonucleotides can preferentially knock down disease-associated transcripts of gene targets relative to wild-type (e.g., non-disease-associated) transcripts. In some embodiments, the disease-associated transcripts can contain disease-associated mutations or repeat expansions.

[0016] In some embodiments, the present disclosure provides oligonucleotides having an asymmetric format and comprising any structure or format (or portion thereof) described herein, optional additional chemical moieties (including but not limited to carbohydrate moieties and targeting moieties), stereochemistry or pattern of stereochemistry, internucleotide linkage or pattern of internucleotide linkages; sugar modifications or pattern of sugar modifications; base modifications or pattern of base modifications.

[0017] In some embodiments, the present disclosure provides methods for reducing the level of a nucleic acid or a product encoded thereby, the methods comprising contacting the nucleic acid with a provided oligonucleotide or composition thereof, wherein the base sequence of the oligonucleotide is complementary to the base sequence of the nucleic acid or a portion thereof. In some embodiments, the present disclosure provides methods for treating and / or preventing and / or curing various associated conditions, disorders and / or diseases in a subject, the methods comprising administering to the subject a provided oligonucleotide or composition thereof. [Brief explanation of the drawings]

[0018] [Figure 1A] Figure 1 presents certain provided formats of oligonucleotides having asymmetric formats as non-limiting examples: Figure 1A presents non-limiting examples of sugar modifications and / or patterns thereof on the first and second wings of oligonucleotides having asymmetric formats; [Figure 1B] Figure 1 presents certain provided formats of oligonucleotides having asymmetric formats as non-limiting examples, and Figure 1B presents non-limiting examples of sugar modifications and / or patterns thereof on the first and second wings of oligonucleotides having asymmetric formats. [Figure 1C] Figure 1 provides certain exemplary formats of oligonucleotides having asymmetric formats as non-limiting examples, and Figure 1C provides non-limiting examples of internucleotide linkages in the first and second wings of oligonucleotides having asymmetric formats. [Figure 1D] Figure 1 presents certain provided formats of oligonucleotides having asymmetric formats as non-limiting examples. Figure 1D provides a legend for Figures 1A, 1B, and 1C. [Figure 2]Figure 2 shows exemplary cleavage data for oligonucleotides containing specific stereochemical patterns. The arrows indicate the observed cleavage sites. As demonstrated, oligonucleotides containing specific stereochemical patterns can direct cleavage to selected sites. In some embodiments, as shown herein, when using chiral-controlled oligonucleotide compositions of oligonucleotides containing specific stereochemical patterns, cleavage occurs primarily at one site. [Figure 3] FIG. 3 provides exemplary cleavage data with the provided oligonucleotides. [Figure 4] Figure 4 shows exemplary C9orf72 transcripts. The V3, V2, and V1 transcripts produced from healthy and diseased C9orf72 alleles are shown, where the diseased allele contains a hexanucleotide repeat expansion (denoted by a horizontal bar, (GGGGCC)30+). Downward arrows indicate the locations of several exemplary C9orf72 oligonucleotides. DETAILED DESCRIPTION OF THE INVENTION

[0019] definition As used herein, the following definitions shall apply unless otherwise stated. For purposes of this disclosure, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed. In addition, general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5th Ed., Ed.: Smith, Mb and March, J., John Wiley & Sons, New York: 2001.

[0020] Aliphatic: As used herein, "aliphatic" means a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more units of unsaturation, or a substituted or unsubstituted monocyclic, bicyclic, or polycyclic hydrocarbon ring (but not aromatic) that is fully saturated or contains one or more units of unsaturation, or a combination thereof. In some embodiments, an aliphatic group contains 1-50 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-20 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-10 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-9 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-8 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-7 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-6 aliphatic carbon atoms. In yet other embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms, and in still other embodiments, aliphatic groups contain 1, 2, 3, or 4 aliphatic carbon atoms. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups, and hybrids such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl.

[0021] Alkenyl: As used herein, the term "alkenyl" refers to an alkyl group, as defined herein, having one or more double bonds.

[0022] Alkyl: As used herein, the term "alkyl" is given its ordinary meaning in the art and includes saturated aliphatic groups, including straight-chain alkyl groups, branched alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups. In some embodiments, an alkyl has 1-100 carbon atoms. In some embodiments, a straight-chain or branched-chain alkyl has about 1-20 carbon atoms in its backbone (e.g., C1-C for straight chain). 20 , C2 to C in the case of branched chains 20) or about 1-10. In some embodiments, cycloalkyl rings have from about 3-10 carbon atoms in their ring structure (and such rings are monocyclic, bicyclic or polycyclic) or about 5, 6 or 7 carbons in the ring structure. In some embodiments, alkyl groups can be lower alkyl groups, where the lower alkyl group has from 1-4 carbon atoms (e.g., C1-C4 for a straight chain lower alkyl).

[0023] Alkynyl: As used herein, the term "alkynyl" refers to an alkyl group, as defined herein, having one or more triple bonds.

[0024] Approximately: As used herein, the term "approximately" or "about" in reference to a numerical value is generally interpreted to include values ​​within 5%, 10%, 15%, or 20% (more or less) of that value in either direction, unless otherwise stated or otherwise clear from the context (except where such value falls below 0% or exceeds 100% of the possible value). In some embodiments, when the term "about" is used in reference to dosage, it means ±5 mg / kg / day.

[0025] Aryl: As used herein, the term "aryl," used alone or as part of a larger moiety such as "aralkyl," "aralkoxy," or "aryloxyalkyl," refers to a monocyclic, bicyclic, or polycyclic ring system having a total of 5 to 30 ring members, wherein at least one ring of the system is aromatic. In some embodiments, an aryl group refers to a monocyclic, bicyclic, or polycyclic ring system having a total of 5 to 14 ring members, wherein at least one ring of the system is aromatic and wherein the rings of the system each contain 3 to 7 ring members. In some embodiments, an aryl group is a biaryl group. The term "aryl" may be used interchangeably with the term "aryl ring." In some embodiments of the present invention, "aryl" refers to an aromatic ring system, including, but not limited to, phenyl, biphenyl, naphthyl, binaphthyl, anthracyl, and the like, which may contain one or more substituents. Also included within the scope of the term "aryl," as used herein, are groups in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthymidyl, phenanthridinyl, or tetrahydronaphthyl.

[0026] Equivalent: The term "equivalent" is used herein to describe two (or more) sets of conditions or circumstances that are sufficiently similar to one another to permit a comparison of the results obtained or the phenomena observed. In some embodiments, equivalent sets of conditions or circumstances are characterized by a plurality of substantially identical characteristics and one or a small number of dissimilar characteristics. One skilled in the art will understand that sets of conditions are equivalent to one another when they are characterized by a sufficient number and type of substantially identical characteristics to provide a basis for a reasonable conclusion that differences in the results obtained or phenomena observed under the various sets of conditions or circumstances are attributable to or indicative of differences in the dissimilar characteristics.

[0027] Alicyclic: The terms "alicyclic," "carbocycle," "carbocyclyl," "carbocyclic group," and "carbocycle" are used interchangeably and, as used herein, unless otherwise specified, refer to a saturated or partially unsaturated, but non-aromatic, cycloaliphatic monocyclic, bicyclic, or polycyclic ring system, as described herein, having 3 to 30 ring members. Alicyclic groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl, norbornyl, adamantyl, and cyclooctadienyl. In some embodiments, an alicyclic group has 3 to 6 carbons. In some embodiments, an alicyclic group is saturated and cycloalkyl. The term "alicyclic" can also include an alicyclic ring fused to one or more aromatic or non-aromatic rings, such as decahydronaphthyl or tetrahydronaphthyl. In some embodiments, an alicyclic group is bicyclic. In some embodiments, an alicyclic group is tricyclic. In some embodiments, an alicyclic group is polycyclic. In some embodiments, "alicyclic" refers to a C3-C6 monocyclic hydrocarbon or C8-C6 alkylene group that is fully saturated or contains one or more units of unsaturation, but is not aromatic, and has a single point of attachment to the rest of the molecule. 10 Bicyclic or polycyclic hydrocarbons, or C9-C6 rings that are fully saturated or contain one or more unsaturated units, but are not aromatic, and have a single point of attachment to the rest of the molecule. 16 Refers to polycyclic hydrocarbons.

[0028] Heteroaliphatic: The term "heteroaliphatic," as used herein, is given its ordinary meaning in the art and refers to an aliphatic group, as described herein, in which one or more carbon atoms are independently replaced with one or more heteroatoms (e.g., oxygen, nitrogen, sulfur, silicon, phosphorus, etc.). In some embodiments, one or more units selected from C, CH, CH, and CH are independently replaced with one or more heteroatoms (including oxidized and / or substituted forms thereof). In some embodiments, a heteroaliphatic group is a heteroalkyl. In some embodiments, a heteroaliphatic group is a heteroalkenyl.

[0029] Heteroalkyl: The term "heteroalkyl," as used herein, is given its ordinary meaning in the art and refers to an alkyl group, as described herein, in which one or more carbon atoms are independently replaced with one or more heteroatoms (e.g., oxygen, nitrogen, sulfur, silicon, phosphorus, etc.). Examples of heteroalkyl groups include, but are not limited to, alkoxy, poly(ethylene glycol)-, alkyl-substituted amino, tetrahydrofuranyl, piperidinyl, morpholinyl, and the like.

[0030] Heteroaryl: The terms "heteroaryl" and "heteroal-," as used herein, alone or as part of a larger moiety (e.g., "heteroaralkyl" or "heteroaralkoxy"), refer to a monocyclic, bicyclic, or polycyclic ring system having a total of 5 to 30 ring members, where at least one ring of the system is aromatic and at least one aromatic ring atom is a heteroatom. In some embodiments, a heteroaryl group has 5 to 10 ring atoms (i.e., monocyclic, bicyclic, or polycyclic), and in some embodiments, 5, 6, 9, or 10 ring atoms. In some embodiments, a heteroaryl group has 6, 10, or 14 π electrons shared within the cyclic array; and in addition to the carbon atoms, there are 1 to 5 heteroatoms. Heteroaryl groups include, but are not limited to, thienyl, furanyl, pyronyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. In some embodiments, a heteroaryl is a heterobiaryl group such as bipyridyl. The terms "heteroaryl" and "heteroar-," as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl, alicyclic, or heterocyclyl rings, and the bonding group or point of attachment is on the heteroaromatic ring. Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Heteroaryl groups can be monocyclic, bicyclic, or polycyclic. The term "heteroaryl" can be used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaromatic," all of which include optionally substituted rings.The term "heteroaralkyl" refers to an alkyl group substituted with a heteroaryl group, where the alkyl and heteroaryl portions independently are optionally substituted.

[0031] Heteroatom: The term "heteroatom," as used herein, means an atom that is not carbon or hydrogen. In some embodiments, a heteroatom is boron, oxygen, sulfur, nitrogen, phosphorus, or silicon (any oxidized form of nitrogen, sulfur, phosphorus, or silicon; the quaternized form of any basic or substitutable nitrogen of a heterocycle (e.g., N, as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR + (such as in the case of N-substituted pyrrolidinyl); etc.).

[0032] Heterocycle: As used herein, the terms "heterocycle," "heterocyclyl," "heterocyclic group," and "heterocycle" are used interchangeably herein and refer to a monocyclic, bicyclic, or polycyclic ring moiety (e.g., 3-30 membered) that is saturated or partially unsaturated and has one or more heteroatom ring atoms. In some embodiments, a heterocyclyl group is a stable 5- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated and has one or more, preferably 1-4, heteroatoms (as defined above) in addition to carbon atoms. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur, and nitrogen, the nitrogen can be N (such as in 3,4-dihydro-2H-pyrrolyl), NH (such as in pyrrolidinyl), or +It can be NR (such as in the case of N-substituted pyrrolidinyl). The heterocycle can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure, and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms "heterocycle," "heterocyclyl," "heterocyclyl ring," "heterocyclic group," "heterocyclic moiety," and "heterocyclic radical" are used interchangeably herein and also include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or alicyclic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl. Heterocyclyl groups can be monocyclic, bicyclic, or polycyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by a heterocyclyl, where the alkyl and heterocyclyl portions independently are optionally substituted.

[0033] In vitro: As used herein, the term "in vitro" refers to events that take place not within a living organism (e.g., an animal, plant, and / or microorganism), but in an artificial environment, such as a test tube or reaction vessel, cell culture.

[0034] In vivo: As used herein, the term "in vivo" refers to events that take place within an organism (e.g., an animal, a plant, and / or a microorganism).

[0035] Optionally substituted: As described herein, compounds of the present disclosure, such as oligonucleotides, may contain optionally substituted and / or substituted moieties. In general, the term "substituted," whether preceded by the term "optionally," means that one or more hydrogens of the specified moiety are replaced with a suitable substituent. Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each of the group's substitutable positions, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents may be the same or different at all positions. In some embodiments, an optionally substituted group is unsubstituted. Combinations of substituents contemplated by the present disclosure are preferably those that result in the formation of stable or chemically feasible compounds. The term "stable," as used herein, refers to a compound that remains substantially unchanged when subjected to conditions that permit its production, detection, and, in certain embodiments, its recovery, purification, and use for one or more purposes disclosed herein.

[0036] Suitable monovalent substituents on substitutable atoms, e.g., suitable carbon atoms, are independently: halogen; -(CH) 0~4 R 〇 ;-(CH2) 0~4 OR 〇 ;-O(CH2) 0~4 R 〇 , -O-(CH2) 0~4 C(O)OR 〇 ;-(CH2) 0~4 CH(OR 〇 )2;-(CH2) 0~4 Ph(R 〇 -(CH2) 0~4 O(CH2) 0~1 Ph(R 〇 -CH=CHPh(R 〇 -(CH2) 0~4 O(CH2) 0~1 -pyridyl (R 〇 -NO2; -CN; -N3; ​​-(CH2) 0~4 N(R〇 )2;-(CH2) 0~4 N(R 〇 )C(O)R 〇 ;N(R 〇 )C(S)R 〇 ;-(CH2) 0~4 N(R 〇 )C(O)NR 〇 2;-N(R 〇 )C(S)NR 〇 2;-(CH2) 0~4 N(R 〇 )C(O)OR 〇 ;-N(R 〇 )N(R 〇 )C(O)R 〇 ;-N(R 〇 )N(R 〇 )C(O)NR2;-N(R 〇 )N(R 〇 )C(O)OR 〇 ;-(CH2) 0~4 C(O)R 〇 ;-C(S)R 〇 ;-(CH2) 0~4 C(O)OR 〇 ;-(CH2) 0~4 C(O)SR 〇 ;-(CH2) 0~4 C(O)SiR 〇 3;-(CH2) 0~4 CO(O)R 〇 ;-OC(O)(CH2) 0~4 SR 〇 、-SC(S)SR 〇 ;-(CH2) 0~4 SC(O)R 〇 ;-(CH2) 0~4 C(O)NR 〇 2;-C(S)NR 〇 2;-C(S)SR 〇 ;-SC(S)SR 〇 、-(CH2) 0~4 OC(O)NR 〇 2;-C(O)N(OR 〇 )R 〇 ;-C(O)C(O)R 〇 ;-C(O)CH2C(O)R 〇 ;-C(NOR 〇 )R 〇;-(CH2) 0~4 SSR 〇 ;-(CH2) 0~4 S(O)2R 〇 ;-(CH2) 0~4 S(O)2OR 〇 ;-(CH2) 0~4 OS(O)2R 〇 ;-S(O)2NR 〇 2;-(CH2) 0~4 S(O)R 〇 ;-N(R 〇 )S(O)2NR 〇 2;-N(R 〇 )S(O)2R 〇 ;-N(OR 〇 )R 〇 ;-C(NH)NR 〇 2;-Si(R 〇 )3;-OSi(R 〇 )3;-B(R 〇 )2;-OB(R 〇 )2;-OB(OR 〇 )2;-P(R 〇 )2;-P(OR 〇 )2;-OP(R 〇 )2;-OP(OR 〇 )2;-P(O)(R 〇 )2;-P(O)(OR 〇 )2;-OP(O)(R 〇 )2;-OP(O)(OR 〇 )2;-OP(O)(OR 〇 )(SR 〇 );-SP(O)(R 〇 )2;-SP(O)(OR 〇 )2;-N(R 〇 )P(O)(R 〇 )2;-N(R 〇 )P(O)(OR 〇 )2;-P(R 〇 )2[B(R 〇 )3];-P(OR 〇 )2[B(R 〇 )3];-OP(R 〇 )2[B(R 〇 )3];-OP(OR 〇 )2[B(R 〇 )3];-(C 1~4Linear or branched alkylene)ON(R 〇 )2; or -(C 1~4 Linear or branched alkylene)C(O)ON(R 〇 )2, where each R 〇 may be substituted as defined below and independently represent hydrogen, C1-C 20 Aliphatic, C1-C with 1 to 5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon, and phosphorus 20 Heteroaliphatic, -CH2-(C 6~14 aryl), -O(CH2) 0~1 (C 6~14 aryl), -CH2- (5-14 membered heteroaryl ring), a 5-20 membered monocyclic, bicyclic or polycyclic, saturated, partially unsaturated or aryl ring having 0-5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon and phosphorus, or, notwithstanding the above definitions, R 〇 two independent occurrences of together with their intervening atoms form a 5-20 membered monocyclic, bicyclic or polycyclic, saturated, partially unsaturated or aryl ring having 0-5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon and phosphorus, which can be substituted as defined below.

[0037] R 〇 (or R 〇 Suitable monovalent substituents for the ring formed by two independent occurrences of -(CH) together with their intervening atoms are independently halogen, -(CH) 0~2 R · ,-(Halo R · ), -(CH2) 0~2 OH, -(CH2) 0~2 OR · , -(CH2) 0~2 CH(OR · )2;-O(HaloR · ), -CN, -N3, -(CH2) 0~2 C(O)R · , -(CH2) 0~2 C(O)OH, -(CH2) 0~2 C(O)OR · , -(CH2) 0~2 SR · , -(CH2)0~2 SH, -(CH2) 0~2 NH2, -(CH2) 0~2 NHR · , -(CH2) 0~2 NR · 2, -NO2, -SiR · 3. -OSiR · 3. -C(O)SR · , -(C 1~4 Linear or branched alkylene)C(O)OR · or -SSR · (where each R · is unsubstituted or, if preceded by "halo", is substituted with only one or more halogens), and independently C1-C4 aliphatic, -CH2Ph, -O(CH2) 0~1 R is a 5-6 membered saturated, partially unsaturated, or aryl ring having Ph and 0-4 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon, and phosphorus. 〇 Suitable divalent substituents for saturated carbon atoms of include ═O and ═S.

[0038] For example, suitable divalent substituents for suitable carbon atoms are, independently, the following: =O, =S, =NNR * 2, =NNHC(O)R * , =NNHC(O)OR * , =NNHS(O)2R * , =NR * , =NOR * , -O(C(R * 2)) 2~3 O- or -S(C(R * 2)) 2~3 S-, where R * Each independent occurrence of is selected from hydrogen, C1-C6 aliphatic, which may be substituted as defined below, and an unsubstituted 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. Suitable divalent substituents attached to adjacent substitutable carbons of an "optionally substituted" group include -O(CR * 2) 2~3 O-, where R *is selected from hydrogen, C1-C6 aliphatic which may be substituted as defined below, and an unsubstituted 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms selected from nitrogen, oxygen, and sulfur.

[0039] R * Suitable substituents for the aliphatic groups are independently halogen, R · ,-(Halo R · ), -OH, -OR · , -O(HaloR · ), -CN, -C(O)OH, -C(O)OR · , -NH2, NHR · , -NR · 2 or -NO2 (each R · is unsubstituted or, if preceded by "halo", is substituted with one or more halogens only) and independently C1-C4 aliphatic, -CH2Ph, -O(CH2) 0~1 Ph or independently is a 5-6 membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms selected from nitrogen, oxygen and sulfur.

[0040] Oral: The phrases "oral administration" and "orally administered," as used herein, have their art-understood meanings and refer to administration of a compound or composition by mouth.

[0041] Parenteral: The phrases "parenteral administration" and "parenterally administered," as used herein, have their art-understood meaning and refer to modes of administration other than enteral and topical administration, usually by injection, including, but not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraocular, intracardiac, intradermal, intraperitoneal, intratracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0042] Partially unsaturated: As used herein, the term "partially unsaturated" refers to a ring moiety that contains at least one double or triple bond. The term "partially unsaturated" is intended to encompass rings with multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties, as defined herein.

[0043] Pharmaceutical composition: As used herein, the term "pharmaceutical composition" refers to an active agent formulated together with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in a unit dose suitable for administration in a treatment regimen that, when administered to a relevant population, exhibits a statistically significant probability of achieving a predetermined therapeutic effect. In some embodiments, a pharmaceutical composition may be specifically formulated for administration in a solid or liquid dosage form, including those designed for the following administrations: oral administration, e.g., drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., buccal, sublingual, and those targeted for systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, e.g., subcutaneous, intramuscular, intravenous, or epidural injection as a sterile solution or suspension or sustained-release formulation; topical application, e.g., creams, ointments, or controlled-release patches or sprays applied to the skin, lungs, or oral cavity; vaginal or rectal administration, e.g., as a pessary, cream, or foam; sublingual; ophthalmic; transdermal; or intranasal, pulmonary, and other mucosal surfaces.

[0044] Pharmaceutically acceptable: As used herein, the phrase "pharmaceutically acceptable" refers to compounds, materials, compositions and / or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other impairment or complication, commensurate with a reasonable benefit / risk ratio.

[0045] Pharmaceutically acceptable carrier: As used herein, the term "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent encapsulant, that is involved in carrying or transporting a compound of interest from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Pharmaceutically acceptable carriers include: sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives, for example, sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; talc; excipients such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; pH buffer solutions; polyesters; polycarbonates and / or polyanhydrides; and other non-toxic compatible substances used in pharmaceutical formulations.

[0046] Pharmaceutically acceptable salts: As used herein, the term "pharmaceutically acceptable salts" refers to salts of compounds that are suitable for use in connection with pharmaceutical preparations, i.e., salts that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, and that are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are known in the art. For example, S. M. Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66:1-19 (1977). In some embodiments, pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts of amino groups formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by other methods used in the art, such as ion exchange. In some embodiments, pharmaceutically acceptable salts include, but are not limited to, adipate, arginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfate, formate, fumarate, glucoheptanoate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxybenzoate ... -hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like.In some embodiments, provided compounds contain one or more acidic groups, e.g., oligonucleotides, and pharmaceutically acceptable salts are alkali, alkaline earth metal, or ammonium (e.g., ammonium salts of N(R)3, where each R is independently as defined and described in this disclosure) salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. In some embodiments, the pharmaceutically acceptable salt is a sodium salt. In some embodiments, the pharmaceutically acceptable salt is a potassium salt. In some embodiments, the pharmaceutically acceptable salt is a calcium salt. In some embodiments, pharmaceutically acceptable salts include amine cations formed with counterions such as non-toxic ammonium, quaternary ammonium, and halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyls having 1 to 6 carbon atoms, sulfonates, and arylsulfonates, where appropriate. In some embodiments, provided compounds contain more than one acidic group; for example, provided oligonucleotides can contain two or more acidic groups (e.g., at natural phosphate linkages and / or modified internucleotide linkages). In some embodiments, a pharmaceutically acceptable salt, or generally a salt of such a compound, contains two or more cations, which may be the same or different. In some embodiments, in a pharmaceutically acceptable salt (or generally, a salt), all ionized hydrogens in an acidic group are replaced with cations. In some embodiments, a pharmaceutically acceptable salt is a sodium salt of a provided oligonucleotide. In some embodiments, a pharmaceutically acceptable salt is a sodium salt of a provided oligonucleotide, and each acidic phosphate group is present as a salt form (all sodium salts). In some embodiments, a pharmaceutically acceptable salt is a calcium salt of a provided oligonucleotide.

[0047] Protecting Group: The term "protecting group," as used herein, is known in the art and includes those described in detail in Protecting Groups in Organic Synthesis, T.W. Greene and P.G.M. Butts, 3rd edition, John Wiley & Sons, 1999 (incorporated herein by reference in its entirety). It also includes protecting groups specifically designed for nucleoside and nucleotide chemistry as described in Current Protocols in Nucleic Acid Chemistry, edited by Serge L. Beaucage et al. 06 / 2012 (Chapter 2 is incorporated herein by reference in its entirety). Suitable amino protecting groups include methyl carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methyl carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc), 2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate (Teo c), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethyl carbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC), 1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc), 1-(3,5-di-t-biphenylcarbamate)-1-methylethyl carbamate (t-Bumeoc), 2-(2'- and 4'-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-Dicyclohexylcarboxamido)ethyl carbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropyl allyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyl dithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz), p-nitro p-Toluenesulfonylmethylcarbamate, p-bromobenzylcarbamate, p-chlorobenzylcarbamate, 2,4-dichlorobenzylcarbamate, 4-methylsulfinylbenzylcarbamate (Msz), 9-anthrylmethylcarbamate, diphenylmethylcarbamate, 2-methylthioethylcarbamate, 2-methylsulfonylethylcarbamate, 2-(p-toluenesulfonyl)ethylcarbamate, [2-(1,3-dithianyl)]methylcarbamate (Dmoc), 4-methylthiophenylcarbamate (Mtpc), 2,4-dimethyl-thiamine o-phenylcarbamate (Bmpc), 2-phosphinoethyl carbamate (Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc), 1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate, 2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methyl carbamate, phenothiazinyl-(10)-carbonyl derivatives, N'-p-toluenesulfonylaminocarbonyl derivatives, N'-phenylaminothiocarbonyl derivatives, t-amyl carbamate, S-benzylthiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate, 2,2-Dimethoxycarbonylvinylcarbamate, o-(N,N-dimethyl-carboxamido)benzylcarbamate, 1,1-dimethyl-3-(N,N-dimethyl-carboxamido)propylcarbamate, 1,1-dimethyl-propynylcarbamate, di(2-pyridyl)methylcarbamate, 2-furanylmethylcarbamate, 2-iodoethylcarbamate, isobornylcarbamate, isobutylcarbamate, isonicotylcarbamate, p-(p'-methoxyphenylazo)benzylcarbamate, 1-methylcyclobutylcarbamate, 1- Methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate, 1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate, 1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate, 4-(trimethylammonium)benzyl carbamate, 2,4,6-trimethylbenzyl carbamate benzyl carbamate, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivatives, benzamide, p-phenylbenzamide, o-nitrophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide, (N'-dithiobenzyloxycarbonylamino)acetamide, 3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenoxy)acetamide Nyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide, 2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine derivatives, o-nitrobenzamide, o-(benzoyloxymethyl)benzamide, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1,4,4-Tetramethyldisilylazacyclopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine, N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyrrolidone-3-yl)amine, quaternary Ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm), N-2-picolylamino N'-oxide, N-1,1-dimethylthiomethyleneamine, N-benzylideneamine, Np-methoxy Benzylideneamine, N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine, N-(N',N'-dimethylaminomethylene)amine, N,N'-isopropylidenediamine, Np-nitrobenzylideneamine, N-salicylideneamine, N-5-chlorosalicylideneamine, N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine, N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine, N-borane derivatives, N-diphenylborine Acid derivatives, N-[phenyl(pentacarbonylchromium- or tungsten)carbonyl]amine, N-copper chelate, N-zinc chelate, N-nitramine, N-nitrosamine, amine N-oxide, diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt), dialkylphosphoramidates, dibenzylphosphoramidates, diphenylphosphoramidates, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-Dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, 3-nitropyridine sulfenamide (Npys), p-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide ( Mte), 4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide (Ms), β-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide, 4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

[0048] Suitable protected carboxylic acids further include, but are not limited to, silyl-, alkyl-, alkenyl-, aryl-, and arylalkyl-protected carboxylic acids. Examples of suitable silyl groups include trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triisopropylsilyl, and the like. Examples of suitable alkyl groups include methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, and tetrahydropyran-2-yl. Examples of suitable alkenyl groups include allyl. Examples of suitable aryl groups include optionally substituted phenyl, biphenyl, or naphthyl. Examples of suitable arylalkyl groups include optionally substituted benzyl (e.g., p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl), and 2- and 4-picolyl.

[0049] Suitable hydroxyl protecting groups include methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl ( MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]phenyl ]-4-Methoxypiperidin-4-yl (CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-trimethyl- methylethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxide, diphenylmethyl, p,p'-Dinitrobenzohydryl, 5-dibenzosuberyl, triphenylmethyl, α-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4'-bromophenacyloxyphenyl)diphenylmethyl, 4,4',4''-tris(4,5-dichlorophthalimidophenyl)methyl, 4,4',4''-tris(levulinoyloxyphenyl)methyl, 4,4',4''-tris(benzoyl hydroxyphenyl)methyl, 3-(imidazol-1-yl)bis(4',4''-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodithiolan-2-yl, benzisothiazolyl S,S-dioxide, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate, benzoyl formate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmeth Oxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxovalerate (levulinate), 4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate, adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl 2,2,2-Trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec), 2-(triphenylphosphonio)ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl vinyl carbonate, alkyl aryl carbonate, alkyl p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzylthiocarbonate, 4-ethoxy-1-naphthothyl carbonate, methyldithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylvalerate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy) Ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6-dichloro-4-methylphenoxyacetic acid, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetic acid, 2,4-bis(1,1-dimethylpropyl)phenoxyacetic acid, chlorodiphenylacetic acid, isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate, o-(methoxycarbonyl)benzoate, α-naphthoic acid, nitrate, alkyl N,N,N',N'-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate, borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts). When protecting 1,2- or 1,3-diol, the protecting group may be methylene acetal, ethylidene acetal, 1-t-butylethylidene ketal, 1-phenylethylidene ketal, (4-methoxyphenyl)ethylidene acetal, 2,2,2-trichloroethylidene acetal, acetonide, cyclopentylidene ketal, cyclohexylidene ketal, cycloheptylidene ketal, benzylidene acetal, p-methoxybenzylidene acetal, 2,4-dimethoxybenzylidene ketal, 3,Examples of suitable hydroxyl groups include 4-dimethoxybenzylidene acetal, 2-nitrobenzylidene acetal, methoxymethylene acetal, ethoxymethylene acetal, dimethoxymethylene orthoester, 1-methoxyethylidene orthoester, 1-ethoxyethylidene orthoester, 1,2-dimethoxyethylidene orthoester, α-methoxybenzylidene orthoester, 1-(N,N-dimethylamino)ethylidene derivatives, α-(N,N'-dimethylamino)benzylidene derivatives, 2-oxacyclopentylidene orthoester, di-t-butylsilylene group (DTBS), 1,3-(1,1,3,3-tetraisopropyldisiloxanylidene) derivatives (TIPDS), tetra-t-butoxydisiloxane-1,3-diylidene derivatives (TBDS), cyclic carbonates, cyclic boronates, ethyl borate, and phenyl borate.

[0050] In some embodiments, the hydroxyl protecting group is acetyl, t-butyl, t-butoxymethyl, methoxymethyl, tetrahydropyranyl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 2-trimethylsilylethyl, p-chlorophenyl, 2,4-dinitrophenyl, benzyl, benzoyl, p-phenylbenzoyl, 2,6-dichlorobenzyl, diphenylmethyl, p-nitrobenzyl, triphenylmethyl(trityl), 4,4′-dimethoxytrityl, trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triphenylsilyl, triisopropylsilyl, benzoyl formate, chloroacetyl, trichloroacetyl, trifluoroacetyl, pivaloyl, 9-fluorenylmethyl carbonate, mesylate, tosylate, triflate, trityl, monomethoxytrityl (MMTr), 4,4′-dimethoxytrityl, (D MTr) and 4,4',4''-trimethoxytrityl (TMTr), 2-cyanoethyl (CE or Cne), 2-(trimethylsilyl)ethyl (TSE), 2-(2-nitrophenyl)ethyl, 2-(4-cyanophenyl)ethyl 2-(4-nitrophenyl)ethyl (NPE), 2-(4-nitrophenylsulfonyl)ethyl, 3,5-dichlorophenyl, 2,4-dimethylphenyl, 2-nitrophenyl, 4-nitrophenyl, 2,4,6-trimethylphenyl, 2-(2-nitrophenyl)ethyl, butylthiocarbonyl, 4,4',4''-tris(benzoyloxy)trityl, diphenylcarbamoyl, levulinyl, 2-(dibromomethyl)benzoyl (Dbmb), 2-(isopropylthiomethoxymethyl)benzoyl (Ptmt), 9-phenylxanthen-9-yl (Pixyl), or 9-(p-methoxyphenyl)xanthin-9-yl (MOX). In some embodiments, each of the hydroxyl protecting groups is independently selected from acetyl, benzyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, and 4,4'-dimethoxytrityl. In some embodiments, the hydroxyl protecting groups are selected from the group consisting of trityl, monomethoxytrityl, and 4,4'-dimethoxytrityl groups.

[0051] In some embodiments, the phosphorus-linked protecting group is a group that is added to a phosphorus linkage (e.g., an internucleotide linkage) throughout oligonucleotide synthesis. In some embodiments, the protecting group is added to the sulfur atom of a phosphorothioate linkage. In some embodiments, the protecting group is added to the oxygen atom of a phosphorothioate internucleotide linkage. In some embodiments, the protecting group is added to the oxygen atom of a phosphate internucleotide linkage. In some embodiments, the protecting group is 2-cyanoethyl (CE or Cne), 2-trimethylsilylethyl, 2-nitroethyl, 2-sulfonylethyl, methyl, benzyl, o-nitrobenzyl, 2-(p-nitrophenyl)ethyl (NPE or Npe), 2-phenylethyl, 3-(N-tert-butylcarboxamido)-1-propyl, 4-oxopentyl, 4-methylthio-1-butyl, 2-cyano-1,1-dimethylethyl, 4-N-methylaminobutyl, 3-(2-pyridyl)-1-propyl, 2-[N-methyl-N-(2-pyridyl)]aminoethyl, 2-(N-formyl, N-methyl)aminoethyl, or 4-[N-methyl-N-(2,2,2-trifluoroacetyl)amino]butyl.

[0052] Sample: As used herein, a sample refers to a particular organism or material obtained therefrom. In some embodiments, a sample is a biological sample obtained or derived from a source of interest, as described herein. In some embodiments, the source of interest includes an organism, such as an animal or a human. In some embodiments, a biological sample includes biological tissue or bodily fluid. In some embodiments, a biological sample is or includes bone marrow; blood; blood cells; ascites; tissue or fine needle biopsy; cell-containing bodily fluids; free-floating nucleic acids; sputum; saliva; urine; cerebrospinal fluid, peritoneal fluid; pleural fluid; feces; lymphatic fluid; gynecological fluids; skin swabs; vaginal swabs; oral swabs; nasal swabs; lavages or washings, such as ductal lavage or bronchoalveolar lavage; aspirates; scrapings; bone marrow specimens; tissue biopsies; surgical specimens; feces, other bodily fluids, secretions, and / or excretions; and / or cells derived therefrom. In some embodiments, a biological sample is or includes cells obtained from an individual. In some embodiments, a sample is a "primary sample" obtained directly from a source of interest by any suitable means. For example, in some embodiments, a primary biological sample is obtained by a method selected from the group consisting of biopsy (e.g., fine needle aspiration or tissue biopsy), surgery, collection of bodily fluids (e.g., blood, lymph, stool, etc.), etc. In some embodiments, as the context will make clear, the term "sample" refers to a preparation obtained by processing a primary sample (e.g., removing one or more components therefrom and / or adding one or more substances thereto), for example, filtration through a semipermeable membrane. Such a "processed sample" may include, for example, nucleic acids or proteins extracted from the sample or obtained by subjecting the primary sample to techniques such as mRNA amplification or reverse transcription, isolation and / or purification of specific components, etc. In some embodiments, the sample is an organism. In some embodiments, the sample is a plant. In some embodiments, the sample is an animal. In some embodiments, the sample is a human. In some embodiments, the sample is a non-human organism.

[0053] Subject: As used herein, the term "subject" or "test subject" refers to any organism to which a provided compound or composition is administered in accordance with the present disclosure, e.g., for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans; insects; worms; etc.) and plants. In some embodiments, the subject may be afflicted with and / or susceptible to a disease, disorder, and / or condition.

[0054] Substantially: As used herein, the term "substantially" refers to the qualitative condition of exhibiting all or nearly all extent or degree of a characteristic or property of interest. A base sequence that is substantially complementary to a second sequence is not identical to the second sequence, but is largely or nearly identical to the second sequence. Additionally, those skilled in the art of biology will understand that biological and chemical phenomena rarely, if ever, reach and / or proceed to completion or achieve or avoid absolute consequences. Thus, the term "substantially" is used herein to capture the potential lack of completeness inherent in many biological and / or chemical phenomena.

[0055] Suffering from: An individual "suffering from" a disease, disorder, and / or condition has been diagnosed with and / or exhibits one or more symptoms of the disease, disorder, and / or condition.

[0056] Susceptible: An individual who is "susceptible" to a disease, disorder, and / or condition is an individual who is at a higher risk than the general population of developing the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition is predisposed to contract the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may not have been diagnosed with the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may exhibit symptoms of the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may not exhibit symptoms of the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition will develop the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may not develop the disease, disorder, and / or condition.

[0057] Systemic: The phrases "systemic administration," "administered systemically," "peripheral administration," and "administered peripherally," as used herein, have their art-understood meaning of referring to administration of a compound or composition so that it enters the recipient's entire body.

[0058] Therapeutic Agent: As used herein, the phrase "therapeutic agent" refers to any agent that has a therapeutic effect and / or induces a desired biological and / or pharmacological effect when administered to a subject. In some embodiments, a therapeutic agent is any substance that can be used to alleviate, ameliorate, relieve, inhibit, prevent, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or characteristics of a disease, disorder, and / or condition.

[0059] Therapeutically effective amount: As used herein, the term "therapeutically effective amount" refers to an amount of a substance (e.g., a therapeutic agent, composition, and / or formulation) that, when administered as part of a treatment regimen, elicits a desired biological response. In some embodiments, a therapeutically effective amount of a substance is an amount that, when administered to a subject suffering from or susceptible to a disease, disorder, and / or condition, is sufficient to treat, diagnose, prevent, and / or delay the onset of the disease, disorder, and / or condition. As will be appreciated by those of skill in the art, the effective amount of a substance can vary depending on factors such as the desired biological endpoint, the substance being delivered, the target cell or tissue, and the like. For example, an effective amount of a compound in a formulation for treating a disease, disorder, and / or condition is an amount that alleviates, ameliorate, relieves, inhibits, prevents, delays the onset of, reduces the severity of, and / or reduces the incidence of, one or more symptoms or characteristics of the disease, disorder, and / or condition. In some embodiments, a therapeutically effective amount is administered in a single dose; in some embodiments, multiple unit doses are required to deliver a therapeutically effective amount.

[0060] Treat: As used herein, the terms "treat," "treatment," or "treating" refer to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or characteristics of a disease, disorder, and / or condition. Treatment may be administered to subjects who do not show signs of the disease, disorder, and / or condition. In some embodiments, treatment may be administered to subjects who show only early signs of the disease, disorder, and / or condition, for the purpose of, for example, reducing the risk of developing pathology associated with the disease, disorder, and / or condition.

[0061] Unsaturated: As used herein, the term "unsaturated" means that a moiety has one or more units of unsaturation.

[0062] Unit dose: As used herein, the phrase "unit dose" refers to an amount administered as a single administration of a pharmaceutical composition and / or administered in a physically discrete unit. In many embodiments, a unit dose comprises a predetermined amount of an active agent. In some embodiments, a unit dose comprises an entire single dose of a drug. In some embodiments, two or more unit doses are administered to achieve a total single administration. In some embodiments, administration of multiple unit doses is necessary or expected to be necessary to achieve the intended effect. A unit dose can be, for example, a predetermined amount of one or more therapeutic agents, a volume of a liquid (e.g., an acceptable carrier) containing a predetermined amount of one or more therapeutic agents in solid form, or a sustained release formulation or drug delivery device containing a predetermined amount of one or more therapeutic agents. It will be understood that a unit dose can be present in a formulation that includes any of a variety of ingredients in addition to a therapeutic agent. For example, acceptable carriers (e.g., pharmaceutically acceptable carriers), diluents, stabilizers, buffers, preservatives, etc., can be included, as described below. Those skilled in the art will understand that in many embodiments, an appropriate total daily dosage of a particular therapeutic agent may comprise a fraction or multiple of a single unit dose, which may be determined by the attending physician within the scope of sound medical judgment. In some embodiments, the specific effective dosage level for any particular subject or organism may vary depending on a variety of factors, including the disorder being treated and the severity of the disorder, the activity of the specific active compound used; the specific composition used; the age, weight, health, sex, and diet of the subject; the time of administration and its excretion rate of the specific active compound used; the duration of treatment; drugs and / or additional therapies used in combination or simultaneously with the specific compound used, and similar factors known in the medical arts.

[0063] Wild-type: As used herein, the term "wild-type" has its art-recognized meaning of referring to an entity having a structure and / or activity that occurs in nature in a "normal" state or situation (as opposed to a mutant, diseased, modified, etc.). Those of skill in the art will understand that wild-type genes and polypeptides often exist in multiple alternative forms (e.g., alleles).

[0064] Nucleic Acid: As used herein, the term "nucleic acid" includes any nucleotide and polymers thereof. As used herein, the term "polynucleotide" refers to a polymeric form of nucleotides of any length, including ribonucleotides (RNA) or deoxyribonucleotides (DNA). These terms refer to the primary structure of the molecule and, thus, include double- and single-stranded DNA and double- and single-stranded RNA. These terms include, as equivalents, analogs of either RNA or DNA made from modified nucleotides and / or modified polynucleotides, such as, but not limited to, methylated, protected, and / or capped nucleotides or polynucleotides. These terms encompass polyribonucleotides or oligoribonucleotides (RNA) and polydeoxyribonucleotides or oligodeoxyribonucleotides (DNA); RNA or DNA derived from nucleobases and / or N-glycosides or C-glycosides of modified nucleobases; nucleic acids derived from sugars and / or modified sugars; and nucleic acids derived from phosphate bridges and / or modified phosphorus internucleotide linkages. The term encompasses nucleic acids containing any combination of nucleobases, modified nucleobases, sugars, modified sugars, phosphate bridges, or modified internucleotide linkages. Examples include, but are not limited to, nucleic acids containing ribose moieties, nucleic acids containing deoxyribose moieties, nucleic acids containing both ribose and deoxyribose moieties, and nucleic acids containing ribose and modified ribose moieties. Unless otherwise specified, the prefix poly- refers to a nucleic acid containing from 2 to about 10,000 nucleotide monomer units, and the prefix oligo- refers to a nucleic acid containing from 2 to about 200 nucleotide monomer units.

[0065] Nucleotide: As used herein, the term "nucleotide" refers to a monomeric unit of a polynucleotide composed of a nucleobase, a sugar, and one or more internucleotide linkages. Natural bases (guanine (G), adenine (A), cytosine (C), thymine (T), and uracil (U)) are derivatives of purines or pyrimidines, although it should be understood that natural and unnatural base analogs are also included. Natural sugars are the pentoses (five-carbon sugars) deoxyribose (forming DNA) or ribose (forming RNA), although it should be understood that natural and unnatural sugar analogs are also included. Nucleotides are linked via internucleotide linkages to form nucleic acids or polynucleotides. Many internucleotide linkages are known in the art (e.g., but not limited to, phosphate, phosphorothioate, boranophosphate, etc.). Artificial nucleic acids include PNAs (peptide nucleic acids), phosphotriesters, phosphorothioates, H-phosphonates, phosphoramidates, boranophosphates, methylphosphonates, phosphonoacetates, thiophosphonoacetates, and other variants of the phosphate backbone of native nucleic acids, such as those described herein. In some embodiments, natural nucleotides contain naturally occurring bases, sugars, and internucleotide linkages. As described herein, in some embodiments, the term "nucleotide" also encompasses structural analogs that are used in place of natural or naturally occurring nucleotides, such as modified nucleotides and nucleotide analogs.

[0066] Modified Nucleotide: The term "modified nucleotide" includes any chemical moiety that is structurally different from a naturally occurring nucleotide but is capable of performing at least one function of a naturally occurring nucleotide. In some embodiments, a modified nucleotide comprises a modification in the sugar, base, and / or internucleotide linkage. In some embodiments, a modified nucleotide comprises a modified sugar, a modified nucleobase, and / or a modified internucleotide linkage. In some embodiments, a modified nucleotide is capable of performing at least one function of a nucleotide, e.g., forming a subunit in a polymer that is capable of base pairing with a nucleic acid comprising at least a complementary base sequence.

[0067] Analog: The term "analog" includes any chemical moiety that is structurally different from a reference chemical moiety or class of chemical moieties, but that is capable of performing at least one function of such reference chemical moiety or class of chemical moieties. As non-limiting examples, a nucleotide analog is structurally different from a nucleotide, but performs at least one function of a nucleotide; a nucleobase analog is structurally different from a nucleobase, but performs at least one function of a nucleobase, etc.

[0068] Nucleoside: The term "nucleoside" refers to a moiety in which a nucleobase or modified nucleobase is covalently linked to a sugar or modified sugar.

[0069] Modified Nucleoside: The term "modified nucleoside" refers to a moiety that is derived from a natural nucleoside or is chemically similar to a natural nucleotide, but contains a chemical modification that distinguishes it from a natural nucleoside. Non-limiting examples of modified nucleosides include those containing base and / or sugar modifications. Non-limiting examples of modified nucleosides include those having a 2' modification on the sugar. Further non-limiting examples of modified nucleosides include abasic nucleosides (nucleobases missing). In some embodiments, the modified nucleoside is capable of at least one function of a nucleoside, for example, forming a moiety in a polymer that is capable of base pairing with a nucleic acid containing at least a complementary base sequence.

[0070] Nucleoside Analog: The term "nucleoside analog" refers to a chemical moiety that is chemically different from a naturally occurring nucleoside but is capable of performing at least one function of a nucleoside. In some embodiments, a nucleoside analog comprises a sugar analog and / or a nucleobase analog. In some embodiments, a modified nucleoside is capable of performing at least one function of a nucleoside, e.g., forming a moiety in a polymer that is capable of base pairing with a nucleic acid containing a complementary base sequence.

[0071] Sugar: The term "sugar" refers to a monosaccharide or polysaccharide in a closed and / or open state. In some embodiments, a sugar is a monosaccharide. In some embodiments, a sugar is a polysaccharide. Sugars include, but are not limited to, ribose, deoxyribose, pentofuranose, pentopyranose, and hexopyranose moieties. As used herein, the term "sugar" also encompasses structural analogs that are used in place of traditional sugar molecules, such as glycols, polymers of which form the backbone of nucleic acid analogs, such as glycol nucleic acids (GNAs). As used herein, the term "sugar" also encompasses structural analogs that are used in place of natural or naturally occurring nucleotides, such as modified sugars and nucleotide sugars.

[0072] Modified sugar: The term "modified sugar" refers to a moiety that can replace a sugar. The modified sugar mimics the spatial arrangement, electronic properties, or any other physicochemical property of a sugar.

[0073] Nucleobase: The term "nucleobase" refers to the portion of a nucleic acid that participates in hydrogen bonding to bind one nucleic acid strand to another complementary strand in a sequence-specific manner. The most common naturally occurring nucleobases are adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T). In some embodiments, a naturally occurring nucleobase is a modified adenine, guanine, uracil, cytosine, or thymine. In some embodiments, a naturally occurring nucleobase is a methylated adenine, guanine, uracil, cytosine, or thymine. In some embodiments, a nucleobase is a "modified nucleobase," e.g., a nucleobase other than adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T). In some embodiments, a modified nucleobase is a methylated adenine, guanine, uracil, cytosine, or thymine. In some embodiments, modified nucleobases mimic the spatial arrangement, electronic properties, or any other physicochemical properties of nucleobases, and retain the hydrogen bonding properties that bind one nucleic acid strand to another in a sequence-specific manner. In some embodiments, modified nucleobases can pair with all five natural bases (uracil, thymine, adenine, cytosine, or guanine) without substantially affecting the melting behavior, recognition by intracellular enzymes, or activity of oligonucleotide double helix strands. As used herein, the term "nucleobase" also includes structural analogs that are used in place of natural or naturally occurring nucleotides, such as modified nucleobases and nucleobase analogs.

[0074] Modified nucleobase: The terms "modified nucleobase," "modified base," and the like refer to a chemical moiety that is chemically different from a nucleobase but can perform at least one function of a nucleobase. In some embodiments, a modified nucleobase is a nucleobase that includes a modification. In some embodiments, a modified nucleobase is capable of performing at least one function of a nucleobase, for example, can form a moiety in a polymer that can base pair with a nucleic acid that includes at least a complementary base sequence.

[0075] Blocking group: The term "blocking group" refers to a group that masks the reactivity of a functional group. The functional group may subsequently be unmasked by removal of the blocking group. In some embodiments, the blocking group is a protecting group.

[0076] Moiety: The term "moiety" refers to a specific segment of a molecule's functionality. A chemical moiety is a commonly recognized chemical entity embedded in or attached to a molecule.

[0077] Solid Support: The term "solid support" refers to any support that allows for the synthesis of nucleic acids. In some embodiments, the term refers to glass or a polymer that is insoluble in the media used in the reaction steps performed to synthesize nucleic acids and that has been derivatized to contain reactive groups. In some embodiments, the solid support is highly cross-linked polystyrene (HCP) or controlled pore glass (CPG). In some embodiments, the solid support is controlled pore glass (CPG). In some embodiments, the solid support is a hybrid support of controlled pore glass (CPG) and highly cross-linked polystyrene (HCP).

[0078] Homology: "Homology" or "identity" or "similarity" refers to sequence similarity between two nucleic acid molecules. Homology and identity can each be determined by comparing a position in each sequence, which can be aligned for purposes of comparison. When an equivalent position in the compared sequences is occupied by the same base, the molecules are identical at that position; when an equivalent site is occupied by an identical or similar (e.g., similar in steric and / or electronic properties) nucleic acid residue, the molecules can be called homologous (similar) at that position. Expression as a percentage of homology / similarity or identity refers to a function of the number of identical or similar nucleic acids at a position shared by the compared sequences. An "unrelated" or "non-homologous" sequence shares less than 40% identity, less than 35% identity, less than 30% identity, or less than 25% identity with a sequence described herein. When comparing two sequences, the absence of residues (amino acids or nucleic acids) or the presence of extra residues also reduces the identity and homology / similarity.

[0079] In some embodiments, the term "homology" refers to a mathematically based comparison of sequence similarity and is used to identify genes with similar functions or motifs. The nucleic acid sequences described herein can be used as a "query sequence" to perform searches against public databases to identify, for example, other family members, related sequences, or homologs. In some embodiments, such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. (1990) J. Mol. Biol. 215:403-10. In some embodiments, BLAST nucleotide searches can be performed with the NBLAST program, score=100, wordlength=12, to obtain nucleotide sequences homologous to the nucleic acid molecules of the present disclosure. In some embodiments, gapped BLAST can be used, as described in Altschul et al. (1997) Nucleic Acids Res. 25(17):3389-3402, to obtain gapped alignments for comparison purposes. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (eg, XBLAST and BLAST) can be used (see www.ncbi.nlm.nih.gov).

[0080] Identity: As used herein, "identity" means the percentage of identical nucleotide residues at corresponding positions in two or more sequences, when the sequences are aligned so that sequence matching is maximized, i.e., taking into account gaps and insertions. Identity can be readily calculated by known methods, including, but not limited to, those described in: (Computational Molecular Biology, Lesk, A.M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D.W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part I, Griffin, A.M., and Griffin, H.G., eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991; and Carillo, H., and Lipman, D., SIAM J. Applied Mathematics, 1999). Math., 48:1073 (1988). Methods to determine identity are designed to maximize the match between the sequences examined. Moreover, methods to determine identity are codified in publicly available computer programs.Computer program methods for determining identity between two sequences include, but are not limited to, the GCG program package (Devereux, J., et al., Nucleic Acids Research 12(1):387(1984)), BLASTP, BLASTN, and FASTA (Altschul, S. F. et al., J. Molec. Biol. 215:403-410(1990) and Altschul et al. Nuc. Acids Res. 25:3389-3402(1997)). BLAST X programs are publicly available from NCBI and other sources (BLAST Manual, Altschul, S., et al., NCBI NLM NIH Bethesda, Md. 20894; Altschul, S., et al., J. Mol. Biol. 215:403-410 (1990)). The well-known Smith-Waterman algorithm may also be used to determine identity.

[0081] Oligonucleotide: The term "oligonucleotide" refers to a polymer or oligomer of nucleotides, which may contain any combination of natural and unnatural nucleobases, sugars, and internucleotide linkages.

[0082] Oligonucleotides can be single-stranded or double-stranded. Single-stranded oligonucleotides can have a double-stranded region (formed by two portions of the single-stranded oligonucleotide), and double-stranded oligonucleotides containing two oligonucleotide strands can have a single-stranded region, for example, in a region where the two oligonucleotide strands are not complementary to each other. Examples of oligonucleotides include, but are not limited to, structural genes, genes including regulatory and termination regions, self-replicating systems such as viral DNA or plasmid DNA, single-stranded and double-stranded RNAi agents and other RNA interference reagents (RNAi agents or iRNA agents), shRNA, antisense oligonucleotides, ribozymes, microRNAs, microRNA mimics, supermirs, aptamers, antimirs, antagomirs, Ul adapters, triplex-forming oligonucleotides, G-quadruplex oligonucleotides, RNA activators, immunostimulatory oligonucleotides, and decoy oligonucleotides.

[0083] Oligonucleotides of the present disclosure can be any of a variety of lengths. In certain embodiments, oligonucleotides can range from about 2 to about 200 nucleotides in length. In various related embodiments, single-stranded, double-stranded, and triple-stranded oligonucleotides can range in length from about 4 to about 10 nucleotides, about 10 to about 50 nucleotides, about 20 to about 50 nucleotides, about 15 to about 30 nucleotides, or about 20 to about 30 nucleotides in length. In some embodiments, oligonucleotides are about 10 to about 40 nucleotides in length. In some embodiments, oligonucleotides are about 9 to about 39 nucleotides in length. In some embodiments, oligonucleotides are at least 4 nucleotides in length. In some embodiments, oligonucleotides are at least 5 nucleotides in length. In some embodiments, oligonucleotides are at least 6 nucleotides in length. In some embodiments, oligonucleotides are at least 7 nucleotides in length. In some embodiments, oligonucleotides are at least 8 nucleotides in length. In some embodiments, oligonucleotides are at least 9 nucleotides in length. In some embodiments, oligonucleotides are at least 10 nucleotides in length. In some embodiments, oligonucleotides are at least 11 nucleotides in length. In some embodiments, the oligonucleotide is at least 12 nucleotides in length. In some embodiments, the oligonucleotide is at least 15 nucleotides in length. In some embodiments, the oligonucleotide is at least 20 nucleotides in length. In some embodiments, the oligonucleotide is at least 25 nucleotides in length. In some embodiments, the oligonucleotide is at least 30 nucleotides in length. In some embodiments, the oligonucleotide is a duplex of complementary strands at least 18 nucleotides in length. In some embodiments, the oligonucleotide is a duplex of complementary strands at least 21 nucleotides in length. In some embodiments, each nucleotide counted in the length independently comprises an optionally substituted nucleobase selected from adenine, cytosine, guanosine, thymine, and uracil.

[0084] Internucleotide linkage: As used herein, the phrase "internucleotide linkage" generally refers to the bond connecting the nucleotide units of an oligonucleotide or nucleic acid. In some embodiments, the internucleotide linkage is a phosphodiester bond (a natural phosphate bond) as present in naturally occurring DNA and RNA molecules. In some embodiments, the internucleotide linkage comprises a modified internucleotide linkage. In some embodiments, the internucleotide linkage is a "modified internucleotide linkage" in which each oxygen atom of the phosphodiester bond is optionally and independently replaced by an organic or inorganic moiety. In some embodiments, such organic or inorganic moieties are selected from, but are not limited to, =S, =Se, =NR', -SR', -SeR', -N(R')2, B(R')3, -S-, -Se-, and N(R')-, where each R' is independently as defined and described in this disclosure. In some embodiments, the internucleotide linkage is a phosphotriester linkage, a phosphorothioate diester linkage, or a phosphotriester linkage. TIFF2026016363000003.tif973, or modified phosphorothioate triester linkages. In some embodiments, the internucleotide linkage is, for example, a PNA (peptide nucleic acid) or PMO (phosphorodiamidate morpholino oligomer) linkage. Those skilled in the art will understand that the internucleotide linkage can exist as an anion or cation at a given pH depending on the presence of an acidic or basic moiety in the linkage.

[0085] Non-limiting examples of modified internucleotide linkages are the modified internucleotide linkages designated s, s1, s2, s3, s4, s5, s6, s7, s8, s9, s10, s11, s12, s13, s14, s15, s16, s17 and s18, as described in WO 2017 / 210647.

[0086] For example, (Rp,Sp)-ATsCs1GA has 1) a phosphorothioate internucleotide bond between T and C; TIFF2026016363000004.tif973; and 2) between C and G The oligonucleotide has a phosphorothioate triester internucleotide linkage having the structure TIFF2026016363000005.tif1273. Unless otherwise specified, the designation Rp / Sp preceding an oligonucleotide sequence describes the conformation of the chiral phosphorus atom in that internucleotide linkage in order from 5' to 3' of the oligonucleotide sequence. For example, in (Rp,Sp)-ATsCs1GA, the phosphorus in the "s" bond between T and C has the Rp conformation, and the phosphorus in the "s1" bond between C and G has the Sp conformation. In some embodiments, "All-(Rp)" or "All-(Sp)" is used to indicate that all chiral phosphorus atoms in the oligonucleotide have the same Rp or Sp conformation, respectively.

[0087] Oligonucleotide Type: As used herein, the phrase "oligonucleotide type" refers to a specific base sequence, pattern of backbone linkages (i.e., pattern of internucleotide linkage types, e.g., phosphate, phosphorothioate, etc.), pattern of backbone chiral centers (i.e., pattern of bond phosphorus stereochemistry (Rp / Sp)), and pattern of backbone phosphorus modifications (e.g., "-XLR" in Formula I). 1 " " group pattern). In some embodiments, oligonucleotides commonly referred to as a "type" are structurally identical to one another.

[0088] Those skilled in the art will appreciate that the synthesis methods of the present disclosure provide a degree of control during oligonucleotide chain synthesis, whereby each nucleotide unit of the oligonucleotide chain can be pre-designed and / or pre-selected to have a specific stereochemistry at the binding phosphorus and / or a specific modification at the binding phosphorus, and / or a specific base and / or a specific sugar. In some embodiments, the oligonucleotide chain is pre-designed and / or pre-selected to have a specific combination of stereocenters at the binding phosphorus. In some embodiments, the oligonucleotide chain is designed and / or pre-determined to have a specific combination of modifications at the binding phosphorus. In some embodiments, the oligonucleotide chain is designed and / or pre-selected to have a specific combination of bases. In some embodiments, the oligonucleotide chain is designed and / or pre-selected to have a specific combination of one or more of the structural characteristics described above. In some embodiments, the present disclosure provides compositions (e.g., chiral controlled oligonucleotide compositions) containing or composed of multiple oligonucleotide molecules. In some embodiments, all of these molecules are the same type of molecule (i.e., structurally identical to each other). In many embodiments, the provided compositions contain multiple oligonucleotides of different types, typically in predetermined relative amounts.

[0089] Chiral control: As used herein, "chiral control" refers to the control of the stereochemical assignment of the chiral linkage phosphorus in a chiral internucleotide linkage within an oligonucleotide. In some embodiments, the control is achieved through chiral elements absent from the sugar and base moieties of the oligonucleotide; for example, in some embodiments, the control is achieved through one or more chiral auxiliary groups during the oligonucleotide preparation process, as exemplified in the present disclosure, which chiral auxiliary is often part of the chiral phosphoramidite used in the oligonucleotide preparation process. In contrast to chiral control, those skilled in the art will understand that conventional oligonucleotide synthesis without a chiral auxiliary cannot control the stereochemistry at the chiral internucleotide linkage when such conventional oligonucleotide synthesis is used to form the chiral internucleotide linkage. In some embodiments, the stereochemical assignment of each chiral linkage phosphorus in a chiral internucleotide linkage within an oligonucleotide is controlled.

[0090] Chirality-controlled oligonucleotide composition: As used herein, the terms "chirality-controlled oligonucleotide composition," "chirality-controlled nucleic acid composition," and the like refer to a composition comprising a plurality of oligonucleotides (or nucleic acids) having 1) a common base sequence, 2) a common pattern of backbone linkages, and 3) a common pattern of backbone phosphorus modifications, wherein the plurality of oligonucleotides (or nucleic acids) have the same stereochemistry at one or more chiral internucleotide linkages (chirality-controlled internucleotide linkages, the chiral linkage phosphorus is Rp or Sp in the composition, rather than random Rp and Sp as in non-chirality-controlled internucleotide linkages). The level of the plurality of oligonucleotides (or nucleic acids) in the chirality-controlled oligonucleotide composition is predetermined / controlled (e.g., stereoselective formation of one or more chiral internucleotide linkages via chirality-controlled oligonucleotide formulations). In some embodiments, about 1% to 100% (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, or about 5%, 10%, 20%, 30% or less) of the total oligonucleotides in the chiral control oligonucleotide composition. , 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%) are the plurality of oligonucleotides.In some embodiments, about 1% to 100% (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%) of all oligonucleotides having a common base sequence, a common pattern of backbone linkages, and a common pattern of backbone phosphorus modifications in the chiral control oligonucleotide composition. 0%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% are the plurality of oligonucleotides. In some embodiments, the predetermined level is between about 1% and 100% (e.g., about 5%) of all oligonucleotides in a composition, or of all oligonucleotides in a composition that have a shared base sequence (e.g., of a plurality of oligonucleotides or of an oligonucleotide type), or of all oligonucleotides in a composition that have a common base sequence, a common pattern of backbone linkages, and a common pattern of backbone phosphorus modifications among a plurality of oligonucleotides, or of all oligonucleotides in a composition that have a common base sequence, a common pattern of base modifications, a common pattern of sugar modifications, a common pattern of internucleotide linkage types, and / or a common pattern of internucleotide linkage modifications. ~100%, 10%~100%, 20%~100%, 30%~100%, 40%~100%, 50%~100%, 60%~100%, 70%~100%, 80%~100%, 90%~100%, 95%~100%, 50%~90% or approximately 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% , 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%.In some embodiments, the plurality of oligonucleotides have the same stereochemistry at about 1 to 50 (e.g., about 1 to 10, 1 to 20, 5 to 10, 5 to 20, 10 to 15, 10 to 20, 10 to 25, 10 to 30, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) chiral internucleotide linkages. In some embodiments, the plurality of oligonucleotides has between about 1% and 100% (e.g., between about 5% and 100%, 10% and 100%, 20% and 100%, 30% and 100%, 40% and 100%, 50% and 100%, 60% and 100%, 70% and 100%, 80% and 100%, 90% and 100%, 95% and 100%, 50% and 90%, or about 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 50%, 50%, 60%, 70%, 80%, 90%, 95%, 50%, 50%, 60%, 70%, 80%, 90%, 95%, 90%, 95%, 50 ... 0%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% have the same stereochemistry. In some embodiments, each chiral internucleotide linkage is a chiral-controlled internucleotide linkage, and the composition is a completely chiral-controlled oligonucleotide composition. In some embodiments, not all of the chiral internucleotide linkages are chiral-controlled internucleotide linkages, and the composition is a partially chiral-controlled oligonucleotide composition. In some embodiments, the chiral-controlled oligonucleotide composition comprises a non-random or controlled level of distinct oligonucleotide or nucleic acid types. For example, in some embodiments, the chiral controlled oligonucleotide composition comprises one oligonucleotide type. In some embodiments, the chiral controlled oligonucleotide composition comprises two or more oligonucleotide types. In some embodiments, the chiral controlled oligonucleotide composition comprises multiple oligonucleotide types.In some embodiments, a chiral controlled oligonucleotide composition is a composition of oligonucleotides of an oligonucleotide type, which composition comprises a non-random or controlled level of a plurality of oligonucleotides of that oligonucleotide type.

[0091] Chirally pure: As used herein, the phrase "chirally pure" is used to describe an oligonucleotide or composition thereof in which all or nearly all (the remainder being impurities) of the oligonucleotide molecules are present in a single diastereomeric type with respect to the linking phosphorus atom.

[0092] Predetermined: Predetermined (or predetermined) means, for example, intentionally selected, non-random, or controlled, as opposed to randomly occurring, randomly, or achieved without control. Upon reading this specification, one of skill in the art will understand that the present disclosure provides techniques that enable the selection of specific chemical and / or stereochemical features to be incorporated into oligonucleotide compositions, as well as the control of the preparation of oligonucleotide compositions having such chemical and / or stereochemical features. Such compositions provided are "predetermined" as described herein. A composition that may contain any oligonucleotide is not a "predetermined" composition because it is a composition that is accidentally produced by a process that is not controlled to intentionally produce specific chemical and / or stereochemical features. In some embodiments, a predetermined composition is a composition that can be intentionally reproduced (e.g., by repeating a controlled process). In some embodiments, a predetermining level of a plurality of oligonucleotides in a composition means that the absolute and / or relative amounts (ratios, percentages, etc.) of the plurality of oligonucleotides in the composition are controlled. In some embodiments, the predetermining level of the plurality of oligonucleotides in the composition is achieved by the preparation of chiral control oligonucleotides.

[0093] Bound phosphorus: As defined herein, the phrase "bound phosphorus" is used to indicate that the particular phosphorus atom involved is a phosphorus atom present in an internucleotide linkage, which phosphorus atom corresponds to the phosphate atom of a phosphodiester internucleotide linkage present in natural DNA and RNA. In some embodiments, the bound phosphorus atom is in a modified internucleotide linkage, where each oxygen atom of the phosphodiester linkage is optionally and independently replaced by an organic or inorganic moiety. In some embodiments, the bound phosphorus atom is P of Formula I. In some embodiments, the bound phosphorus atom is chiral. In some embodiments, the bound phosphorus atom is achiral.

[0094] P-modification: As used herein, the term "P-modification" refers to any modification at the bound phosphorus other than a stereochemical modification. In some embodiments, a P-modification includes the addition, substitution, or removal of a pendant moiety covalently attached to the bound phosphorus. In some embodiments, a "P-modification" ... 1 wherein X, L and R 1 are each independently as defined and described in this disclosure.

[0095] Blockmir: The term "blockmir," as used herein, refers to an oligonucleotide chain in which the pattern of structural features characterizing each individual nucleotide unit is characterized by the presence of at least two consecutive nucleotide units having a common structural feature at their internucleotide phosphorus linkage. A common structural feature refers to a common stereochemistry at the bond phosphorus or a common modification at the bond phosphorus. In some embodiments, at least two consecutive nucleotide units having a common structural feature at the internucleotide phosphorus linkage are referred to as a "block." In some embodiments, provided oligonucleotides are blockmirs.

[0096] In some embodiments, the blockmir is a "stereoblockmir", e.g., at least two consecutive nucleotide units have the same stereochemistry at the bond phosphorus. Such at least two consecutive nucleotide units form a "stereoblock".

[0097] In some embodiments, the blockmir is a "P-modified blockmir," e.g., at least two consecutive nucleotide units have the same modification at the linked phosphorus. Such at least two consecutive nucleotide units form a "P-modified block." For example, (Rp,Sp)-ATsCsGA is a P-modified blockmir because at least two consecutive nucleotide units, i.e., Ts and Cs, have the same P-modification (i.e., both are phosphorothioate diester). In the same oligonucleotide, (Rp,Sp)-ATsCsGA, Ts and Cs form a block, which is a P-modified block.

[0098] In some embodiments, the blockmir is a "linked blockmir," e.g., at least two consecutive nucleotide units have the same stereochemistry and the same modification at the linking phosphorus. At least two consecutive nucleotide units form a "linked block." For example, (Rp,Rp)-ATsCsGA is a linked blockmir because at least two consecutive nucleotide units, i.e., Ts and Cs, have the same stereochemistry (both Rp) and P-modification (both phosphorothioate). In the same oligonucleotide, (Rp,Rp)-ATsCsGA, TsCs form a block, which is a linked block.

[0099] In some embodiments, the blockmir comprises one or more blocks independently selected from a stereoblock, a P-modified block, and a conjugated block, hi some embodiments, the blockmir is a stereoblockmir with respect to one block, and / or a P-modified blockmir with respect to another block, and / or a conjugated blockmir with respect to yet another block.

[0100] Altmer: The term "altmer," as used herein, refers to an oligonucleotide chain in which the pattern of structural features that characterize each individual nucleotide unit thereof is characterized by no two consecutive nucleotide units of the oligonucleotide chain sharing a particular structural feature at the internucleotide phosphorus bond. In some embodiments, an altmer is designed to contain a repeating pattern. In some embodiments, an altmer is designed to not contain a repeating pattern. In some embodiments, a provided oligonucleotide is an altmer.

[0101] In some embodiments, the altmers are "stereoaltomers," eg, no two consecutive nucleotide units have the same stereochemistry at the bond phosphorus.

[0102] In some embodiments, the altmer is a "P-modified altmer," e.g., no two consecutive nucleotide units have the same modification at the bound phosphorus, e.g., All-(Sp)-CAs1GsT, where each bound phosphorus has a different P-modification than the others.

[0103] In some embodiments, the altmer is a "linked altmer," eg, no two consecutive nucleotide units have the same stereochemistry or the same modification at the linking phosphorus.

[0104] Unimer: As used herein, the term "unimer" refers to an oligonucleotide chain in which the pattern of structural features characterizing each individual nucleotide unit is such that all nucleotide units in the chain share at least one common structural feature at the internucleotide phosphorus linkage. A common structural feature refers to a common stereochemistry at the linking phosphorus or a common modification at the linking phosphorus. In some embodiments, a provided oligonucleotide is a unimer.

[0105] In some embodiments, the unimers are "stereounimers," eg, all nucleotide units have the same stereochemistry at the bond phosphorus.

[0106] In some embodiments, the unimer is a "P-modified unimer," eg, all nucleotide units have the same modification at the bound phosphorus.

[0107] In some embodiments, the unimers are "linked unimers," eg, all nucleotide units have the same stereochemistry and the same modification at the linking phosphorus.

[0108] Gapmer: As used herein, the term "gapmer" refers to an oligonucleotide chain characterized in that at least one internucleotide phosphorus linkage of the oligonucleotide chain is a phosphodiester linkage, e.g., as present in naturally occurring DNA or RNA. In some embodiments, two or more internucleotide phosphorus linkages of the oligonucleotide chain are phosphodiester linkages, e.g., as present in naturally occurring DNA or RNA. In some embodiments, provided oligonucleotides are gapmers.

[0109] Skipmer: As used herein, the term "skipmer" refers to a type of gapmer in which every other internucleotide phosphorus bond in an oligonucleotide strand is a phosphodiester bond, such as those present in natural DNA or RNA, and every other internucleotide phosphorus bond in an oligonucleotide strand is a modified internucleotide bond. In some embodiments, provided oligonucleotides are skipmers.

[0110] For purposes of this disclosure, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover.

[0111] The methods and structures described herein with respect to the compounds and compositions of the present disclosure also apply to the pharmaceutically acceptable acid or base addition salts and all stereoisomeric forms of these compounds and compositions.

[0112] Description of Specific Embodiments The present disclosure provides, inter alia, oligonucleotides of specific structural designs comprising base, sugar, and / or internucleotide linkage modifications and / or patterns thereof described herein. In some embodiments, the present disclosure provides compositions of such oligonucleotides, e.g., chiral-controlled oligonucleotide compositions. As shown herein, the provided oligonucleotides and compositions thereof offer numerous advantages, such as greatly improved stability, activity, selectivity, etc. In some embodiments, the present disclosure provides techniques for evaluating and / or using the provided oligonucleotides and compositions thereof. For example, in some embodiments, the present disclosure provides methods for reducing levels of nucleic acids (e.g., transcripts) and / or products encoded thereby (e.g., proteins) using the provided oligonucleotides and / or compositions thereof. In some embodiments, as shown in the present disclosure, the provided techniques (e.g., oligonucleotides, compositions, methods, etc.) result in high efficiency and / or specificity.

[0113] Specific Oligonucleotides and Compositions In some embodiments, the present disclosure provides a region of consecutive nucleotide units: (Nu M )t[(Nu O )n(Nu M )m]y and providing an oligonucleotide comprising: During the ceremony, Each Nu M are independently nucleotide units comprising modified internucleotide linkages; Each Nu O are independently nucleotide units containing a natural phosphate bond; each of t, n, and m is independently 1 to 20; y is a number between 1 and 10.

[0114] In some embodiments, as shown in the present disclosure, such oligonucleotides provide improved properties, such as improved stability and / or activity.

[0115] In some embodiments, y is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, y is 1. In some embodiments, y is 2. In some embodiments, y is 3. In some embodiments, y is 4. In some embodiments, y is 5. In some embodiments, y is 6. In some embodiments, y is 7. In some embodiments, y is 8. In some embodiments, y is 9. In some embodiments, y is 10.

[0116] As defined herein, each Nu M independently comprise a modified internucleotide linkage. In some embodiments, the modified internucleotide linkage is a chiral internucleotide linkage. In some embodiments, the modified internucleotide linkage is of Formula I or a salt thereof. In some embodiments, the modified internucleotide linkage is chiral and of Formula I or a salt thereof. In some embodiments, the modified internucleotide linkage is a phosphorothioate diester linkage. In some embodiments, the modified internucleotide linkage is chiral and chiral-controlled. In some embodiments, each modified internucleotide linkage is chiral-controlled. In some embodiments, Nu M In some embodiments, the internucleotide linkages of the provided oligonucleotides are chiral phosphorothioate diester linkages. M In some embodiments, the Nu of the provided oligonucleotides comprises different types of modified internucleotide linkages. M In some embodiments, the Nu of the provided oligonucleotides comprises a chiral internucleotide linkage having a linking phosphorus atom of different conformation. M In some embodiments, the Nu of the provided oligonucleotides comprises different types of modified internucleotide linkages. M contains a chiral internucleotide linkage with the linking phosphorus atom in a different conformation. MAt least one chiral internucleotide linkage is Sp at its linking phosphorus. In some embodiments, at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 Nu M each independently contains a chiral internucleotide linkage of Sp at its linking phosphorus. M Each chiral internucleotide linkage is Sp at its linking phosphorus. M At least one chiral internucleotide linkage is Rp at its linking phosphorus. M At least one chiral internucleotide bond is Rp at its bond phosphorus and Nu M At least one chiral internucleotide bond in Nu is Sp at its linking phosphorus. M Additional nucleotide units containing modified internucleotide linkages suitable for are known in the art and / or described in this disclosure and can be used in accordance with the present disclosure.

[0117] As defined herein, each Nu O are independently nucleotide units containing a natural phosphate bond. In some embodiments, at least one Nu O is a nucleotide unit containing a natural phosphate bond, where the natural phosphate bond is attached to a carbon atom of the 5'-nucleotide unit and the sugar unit of the nucleotide unit, and the carbon atom is attached to less than two hydrogen atoms. O are independently nucleotide units containing a natural phosphate bond, where the natural phosphate bond is bonded to a carbon atom of the 5'-nucleotide unit and the sugar unit of the nucleotide unit, and the carbon atom is bonded to fewer than two hydrogen atoms. O is -C(R 5S )2- structure, which is Nu O Natural phosphate bonds and Nu O In some embodiments, each Nu O are independently -C(R 5S )2- structure, which is NuO Natural phosphate bonds and Nu O is directly attached to the ring portion of the sugar unit.

[0118] In some embodiments, each Nu O are independently of the formula NI: TIFF2026016363000006.tif1473 or a salt thereof, wherein: BA is C 1~30 Alicyclic, C 6~30 Aryl, C with 1-10 heteroatoms 5~30 Heteroaryl, C with 1-10 heteroatoms 3~30 an optionally substituted group selected from heterocyclyl, natural nucleobase moieties, and modified nucleobase moieties; L O is the natural phosphate bond; L S is -C(R 5S )2- or L; Each R 5S and R S are independently -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -L-R', -L-OR', -L-SR', -LN(R')2, -OL-OR', -OL-SR', or -OLN(R')2; Each L is independently a covalent bond or C 1~30 C having an aliphatic group and 1 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, boron, and silicon 1~30 heteroaliphatic groups, wherein one or more methylene units are optionally and independently selected from C 1~6 Alkylene, C 1~6Alkenylene, -C≡C-, -C(R')2-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O) -, -S(O)2-, -S(O)2N(R')-, -C(O)S-, -C(O)O-, -P(O)(OR')-, -P(O)(SR')-, -P(O)(R')-, -P(O)(NR')-, -P(S)(OR')-, -P(S)(SR')-, -P( one or more carbon atoms are optionally and independently substituted by Cy(S)(R')-, -P(S)(NR')-, -P(R')-, -P(OR')-, -P(SR')-, -P(NR')-, -P(OR')[B(R')3]-, -OP(O)(OR')O-, -OP(O)(SR')O-, -OP(O)(R')O-, -OP(O)(NR')O-, -OP(OR')O-, -OP(SR')O-, -OP(NR')O-, -OP(R')O-, or -OP(OR')[B(R')3]O-; L is replaced by; Ring A S is an optionally substituted 3-20 membered monocyclic, bicyclic or polycyclic ring having 0-10 heteroatoms; s is 0 to 20; each R' is independently -R, -C(O)R, -C(O)OR, or -S(O2)R; Each R is independently -H or C 1~30 Aliphatic, C with 1-10 heteroatoms 1~30 Heteroaliphatic, C 6~30 Aryl, C 6~30 Arylaliphatic, C with 1-10 heteroatoms 6~30 an optionally substituted group selected from arylheteroaliphatic, 5-30 membered heteroaryl having 1-10 heteroatoms, and 3-30 membered heterocyclyl having 1-10 heteroatoms; or two R groups optionally and independently join to form a covalent bond; or two or more R groups on the same atom optionally and independently, together with that atom, form an optionally substituted 3-30 membered monocyclic, bicyclic or polycyclic ring having 0-10 heteroatoms in addition to that atom; or Two or more R groups on two or more atoms optionally and independently, together with their intervening atoms, form an optionally substituted 3-30 membered monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms in addition to the intervening atoms.

[0119] In some embodiments, TIFF2026016363000007.tif1073 is TIFF2026016363000008.tif1473, wherein R 1S , R 2S , R 3S and R 4S Each of the groups independently represents R 5 and as described herein. In some embodiments, TIFF2026016363000009.tif1173 is TIFF2026016363000010.tif1573, wherein R 1S , R 2S , R 3S and R 4S are each independently as described in this disclosure. TIFF2026016363000011.tif1073 is TIFF2026016363000012.tif1373, wherein R 1S , R 2S , R 3S and R 4S each independently as described in this disclosure.

[0120] In some embodiments, L S is -C(R 5S )2-. In some embodiments, one R 5S is -H, and L S is -CHR 5SIn some embodiments, each R 5S is independently R. In some embodiments, -C(R 5S )2- is -C(R)2-. In some embodiments, one R 5S is -H and -C(R 5S )2- is -CHR-. In some embodiments, R is not hydrogen. In some embodiments, R is optionally substituted C 1~6 In some embodiments, R is an optionally substituted C 1~6 In some embodiments, R is alkyl. In some embodiments, R is substituted. In some embodiments, R is unsubstituted. In some embodiments, R is methyl. Examples of other R groups are described broadly in this disclosure. In some embodiments, -C(R 5S )2-C is chiral and is R. In some embodiments, -C(R 5S )2-C is chiral and is S. In some embodiments, -C(R 5S )2- is -(R)-CHMe-. In some embodiments, -C(R 5S )2- is -(S)-CHMe-.

[0121] Specific Oligonucleotide Formats In some embodiments, the present disclosure provides oligonucleotides having an asymmetric format.

[0122] In some embodiments, oligonucleotides having an asymmetric format are capable of mediating a reduction in the level, expression, and / or activity of a target gene or its gene product. In some embodiments, oligonucleotides having an asymmetric format are capable of mediating a reduction in the level, expression, and / or activity of a target gene or its gene product in a cell in vitro.

[0123] In some embodiments, oligonucleotides having an asymmetric format can function via any mechanism, including, but not limited to, steric hindrance or RNase H-based mechanisms. In some embodiments, in the case of steric hindrance, oligonucleotides having an asymmetric format prevent or reduce translation of target mRNA. In some embodiments, in the case of RNase H-based mechanisms, oligonucleotides having an asymmetric format comprise a core containing multiple deoxyribose sugars, which can anneal to a target RNA (such as, but not limited to, a target gene mRNA), thus forming a DNA-RNA duplex strand that acts as a substrate for RNase H, which can cleave the target RNA. In some embodiments, oligonucleotides having an asymmetric format comprise a core flanked on both sides by wings, each of which also anneals to the target RNA. In some embodiments, one or both wings of an oligonucleotide having an asymmetric format can improve target specificity, target binding, stability, deliverability, efficacy, and / or other useful properties of oligonucleotides having an asymmetric format.

[0124] In some embodiments, the provided oligonucleotides comprise or consist of a wing-core-wing, core-wing, or wing-core structure. In some embodiments, one wing is chemically distinct from the core and the other wing. In some embodiments, the wing or core is a block, and the wing-core-wing structure is a blockmir comprising three blocks. In some embodiments, the core is also referred to as a gap. In some embodiments, the wing-core-wing format is also referred to as a wing-gap-wing format. In some embodiments, the core is a gap, in which case each sugar moiety of the core does not contain any of the sugar modifications of the wings. In some embodiments, oligonucleotides having a wing-core-wing structure are also referred to as oligonucleotides having a wing-gap-wing structure. In some embodiments, oligonucleotides having a wing-core-wing structure are also referred to as gapmers.

[0125] In some embodiments, the first wing, second wing, and core may differ in sugar modification or pattern thereof, and / or internucleotide linkage or pattern thereof, and / or internucleotide linkage stereochemistry or pattern thereof.

[0126] In some embodiments, the wing-core-wing motif is represented as "XYZ," where "X" represents the length of the 5' wing, "Y" represents the length of the core, and "Z" represents the length of the 3' wing. In some embodiments, the core is located adjacent to each of the 5' wing and the 3' wing. In some embodiments, X and Z are the same or different lengths and / or have the same or different modifications or modification patterns. In preferred embodiments, Y is 8-15 nucleotides. X, Y, or Z can be any of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, or more nucleotides. In some embodiments, the oligonucleotides described herein have or comprise a wing-core-wing structure, e.g., 5-10-5, 5-10-4, 4-10-4, 4-10-3, 3-10-3, 2-10-2, 5-9-5, 5-9-4, 4-9-5, 5-8-5, 5-8-4, 4-8-5, 5-7-5, 4-7-5, 5-7-4, or 4-7-4.

[0127] In some embodiments, the core comprises at least five contiguous deoxyribose sugars. In some embodiments, the core comprises at least five contiguous deoxyribose sugars, the first wing comprises a first type of sugar modification that is absent in the core, and the second wing comprises a different type of sugar modification that is absent in the core. In some embodiments, the core comprises at least 10 contiguous deoxyribose sugars, the first wing comprises a first type of sugar modification that is absent in the core, and the second wing comprises a different type of sugar modification that is absent in the core. In some embodiments, the core comprises at least 10 contiguous deoxyribose sugars, the first wing is at least five bases in length and comprises a first type of sugar modification that is absent in the core, and the second wing is at least five bases in length and comprises a different type of sugar modification that is absent in the core. In some embodiments, the core comprises at least 10 contiguous deoxyribose sugars, the first wing is at least 5 bases in length and comprises a first type of sugar modification that is absent from the core, and the second wing is at least 5 bases in length and comprises a second type of sugar modification that is absent from the core, wherein the first and second types of sugar modifications are not the same. In some embodiments, the core comprises at least 10 contiguous deoxyribose sugars, the first wing is at least 5 bases in length and comprises a first and second type of sugar modification that is absent from the core, and the second wing is at least 5 bases in length and comprises the first type of sugar modification but not the second type of sugar modification.

[0128] In some embodiments of oligonucleotides having an asymmetric format, the first wing and the second wing are different lengths. In some embodiments of oligonucleotides having an asymmetric format, the first wing and the second wing are the same length.

[0129] In some embodiments, the oligonucleotide has a wing-core-wing structure, where the length of the first wing (in bases) is represented by X, the length of the core is represented by Y, and the length of the second wing is represented by Z, where XYZ can be any of the following: 1-5-1, 1-6-1, 1-7-1, 1-8-1, 1-9-1, 1-10-1, 1-11-1, 1-12-1, 1-13-1, 1-14-1, 1-15-1, 1-16-1, 1-17-1, 1-18-1, 1-19-1, 1-20-1, 1-5-2, 1-6-2, 1-7-2, 1-8-2, 1-9-2, 1-10-2, 1-11-2, 1-12-2, 1-13-2, 1-14-2, 1-15-2, 1-16-2, 1-17-2, 1-18-2, 1-19-2, 1-20-2, 1-5-3, 1-6-3, 1-7-3, 1-8-3, 1-9-3, 1-10-3 , 1-11-3, 1-12-3, 1-13-3, 1-14-3, 1-15-3, 1-16-3, 1-17-3, 1-18-3, 1-19-3, 1-20-3, 1-5-4, 1-6-4, 1-7-4, 1-8-4, 1-9-4, 1-10-4, 1-11-4, 1-12-4, 1-1 3-4, 1-14-4, 1-15-4, 1-16-4, 1-17-4, 1-18-4, 1-19-4, 1-20-4, 1-5-5, 1-6-5, 1-7-5, 1-8-5, 1-9-5, 1-10-5, 1-11-5, 1-12-5, 1-13-5, 1-14-5, 1-15-5 , 1-16-5, 1-17-5, 1-18-5, 1-19-5, 1-20-5, 2-5-1, 2-6-1, 2-7-1, 2-8-1, 2-9-1, 2-10-1, 2-12-1, 2-12-1, 2-13-1, 2-14-1, 2-15-1, 2-16-1, 2-17-1, 2-1 8-1, 2-19-1, 2-20-1, 2-5-2, 2-6-2, 2-7-2, 2-8-2, 2-9-2, 2-10-2, 2-12-2, 2-12-2, 2-13-2, 2-14-2, 2-15-2, 2-16-2, 2-17-2, 2-18-2, 2-19-2, 2-20-2, 2-5-3, 2-6-3, 2-7-3, 2-8-3, 2-9-3, 2-10-3, 2-12-3, 2-12-3, 2-13-3, 2-14-3, 2-15-3, 2-16-3, 2-17-3, 2-18-3, 2-19-3, 2-20-3, 2-5-4, 2-6-4, 2-7-4,2-8-4、2-9-4、2-10-4、2-12-4、2-12-4、2-13-4、2-14-4、2-15-4、2-16-4、2-17-4、2-18-4、2-19-4、2-20-4、2-5-5、2-6-5、2-7-5、2-8-5、2-9-5、2-10-5、2-12-5、2-12-5、2-13-5、2-14-5、2-15-5、2-16-5、2-17-5、2-18-5、2-19-5、2-20-5、3-5-1、3-6-1、3-7-1、3-8-1、3-9-1、3-10-1、3-13-1、3-14-1、3-13-1、3-14-1、3-15-1、3-16-1、3-17-1、3-18-1、3-19-1、3-20-1、3-5-2、3-6-2、3-7-2、3-8-2、3-9-2、3-10-2、3-13-2、3-14-2、3-13-2、3-14-2、3-15-2、3-16-2、3-17-2、3-18-2、3-19-2、3-20-2、3-5-3、3-6-3、3-7-3、3-8-3、3-9-3、3-10-3、3-13-3、3-14-3、3-13-3、3-14-3、3-15-3、3-16-3、3-17-3、3-18-3、3-19-3、3-20-3、3-5-4、3-6-4、3-7-4、3-8-4、3-9-4、3-10-4、3-13-4、3-14-4、3-13-4、3-14-4、3-15-4、3-16-4、3-17-4、3-18-4、3-19-4、3-20-4、3-5-5、3-6-5、3-7-5、3-8-5、3-9-5、3-10-5、3-13-5、3-14-5、3-13-5、3-14-5、3-15-5、3-16-5、3-17-5、3-18-5、3-19-5、3-20-5、4-5-1、4-6-1、4-7-1、4-8-1、4-9-1、4-10-1、4-14-1、4-14-1、4-13-1、4-14-1、4-15-1、4-16-1、4-17-1、4-18-1、4-19-1、4-20-1、4-5-2、4-6-2、4-7-2、4-8-2、4-9-2、4-10-2、4-14-2、4-14-2、4-13-2、4-14-2、4-15-2、4-16-2、4-17-2、4-18-2、4-19-2、4-20-2、4-5-3、4-6-3、4-7-3、4-8-3、4-9-3、4-10-3、4-14-3、4-14-3、4-13-3、4-14-3、4-15-3、4-16-3、4-17-3、4-18-3、4-19-3、4-20-3、4-5-4、4-6-4、4-7-4、4-8-4、4-9-4、4-10-4、4-14-4、4-14-4、4-13-4、4-14-4、4-15-4、4-16-4、4-17-4、4-18-4、4-19-4、4-20-4、4-5-5、4-6-5、4-7-5、4-8-5、4-9-5、4-10-5、4-14-5、4-14-5、4-13-5、4-14-5、4-15-5、4-16-5、4-17-5、4-18-5、4-19-5、4-20-5、5-5-1、5-6-1、5-7-1、5-8-1、5-9-1、5-10-1、5-15-1、5-12-1、5-13-1、5-14-1、5-15-1、5-16-1、5-17-1、5-18-1、5-19-1、5-20-1、5-5-2、5-6-2、5-7-2、5-8-2、5-9-2、5-10-2、5-15-2、5-12-2、5-13-2、5-14-2、5-15-2、5-16-2、5-17-2、5-18-2、5-19-2、5-20-2、5-5-3、5-6-3、5-7-3、5-8-3、5-9-3、5-10-3、5-15-3、5-12-3、5-13-3、5-14-3、5-15-3、5-16-3、5-17-3、5-18-3、5-19-3、5-20-3、5-5-4、5-6-4、5-7-4、5-8-4、5-9-4、5-10-4、5-15-4、5-12-4、5-13-4、5-14-4、5-15-4、5-16-4、5-17-4、5-18-4、5-19-4、5-20-4、5-5-5、5-6-5、5-7-5、5-8-5、5-9-5、5-10-5、5-15-5、5-12-5、5-13-5、5-14-5、5-15-5、5-16-5、5-17-5、5-18-5、5-19-5、5-20-5、1-5-6、1-6-6、1-7-6、1-8-6、1-9-6、1-10-6、1-11-6、1-12-6、1-13-6、1-14-6、1-15-6、1-16-6、1-17-6、1-18-6、1-19-6、1-20-6、2-5-6、2-6-6、2-7-6、2-8-6、2-9-6、2-10-6、2-11-6、2-12-6、2-13-6、2-14-6、2-15-6、2-16-6、2-17-6、2-18-6、2-19-6、2-20-6、3-5-6、3-6-6、3-7-6、3-8-6、3-9-6、3-10-6、3-11-6、3-12-6、3-13-6、3-14-6、3-15-6、3-16-6、3-17-6、3-18-6、3-19-6、3-20-6、4-5-6、4-6-6、4-7-6、4-8-6、4-9-6、4-10-6、4-11-6、4-12-6、4-13-6、4-14-6、4-15-6、4-16-6、4-17-6、4-18-6、4-19-6、4-20-6、5-5-6、5-6-6、5-7-6、5-8-6、5-9-6、5-10-6、5-11-6、5-12-6、5-13-6、5-14-6、5-15-6、5-16-6、5-17-6、5-18-6、5-19-6、5-20-6、6-5-6、6-6-6、6-7-6、6-8-6、6-9-6、6-10-6、6-11-6、6-12-6、6-13-6、6-14-6、6-15-6、6-16-6、6-17-6、6-18-6、6-19-6、6-20-6、7-5-6、7-6-6、7-7-6、7-8-6、7-9-6、7-10-6、7-11-6、7-12-6、7-13-6、7-14-6、7-15-6、7-16-6、7-17-6、7-18-6、7-19-6、7-20-6、1-5-7、1-6-7、1-7-7、1-8-7、1-9-7、1-10-7、1-11-7、1-12-7、1-13-7、1-14-7、1-15-7、1-16-7、1-17-7、1-18-7、1-19-7、1-20-7、2-5-7、2-6-7、2-7-7、2-8-7、2-9-7、2-10-7、2-11-7、2-12-7、2-13-7、2-14-7、2-15-7、2-16-7、2-17-7、2-18-7、2-19-7、2-20-7、3-5-7、3-6-7、3-7-7、3-8-7、3-9-7、3-10-7、3-11-7、3-12-7、3-13-7、3-14-7、3-15-7、3-16-7、3-17-7、3-18-7、3-19-7、3-20-7、4-5-7、4-6-7、4-7-7、4-8-7、4-9-7、4-10-7、4-11-7、4-12-7、4-13-7、4-14-7、4-15-7、4-16-7、4-17-7、4-18-7、4-19-7、4-20-7、5-5-7、5-6-7、5-7-7, 5-8-7, 5-9-7, 5-10-7, 5-11-7, 5-12-7, 5-13-7, 5-14-7, 5-15-7, 5-16-7, 5-17-7, 5-18-7, 5-19-7, 5-20-7, 6-5-7, 6-6-7, 6-7-7, 6-8-7, 6-9-7, 6-10-7, 6-11-7, 6-12-7, 6-13-7, 6- 14-7, 6-15-7, 6-16-7, 6-17-7, 6-18-7, 6-19-7, 6-20-7, 7-5-7, 7-6-7, 7-7-7, 7-8-7, 7-9-7, 7-10-7, 7-11-7, 7-12-7, 7-13-7, 7-14-7, 7-15-7, 7-16-7, 7-17-7, 7-18-7, 7-19-7 or 7-20-7.

[0130] As described herein, the core and wings can have various lengths. In some embodiments, the core comprises 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more nucleobases. In some embodiments, the wings comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more nucleobases. In some embodiments, the wings comprise 2, 3, 4, 5, 6, 7, 8, 9, or 10 or fewer nucleobases. In some embodiments, in the case of a wing-core-wing structure, both wings are the same length, e.g., 5 nucleobases. In some embodiments, the two wings are different lengths. In some embodiments, the core is 40%, 45%, 50%, 60%, 70%, 80%, or 90% or more of the total oligonucleotide length, as measured by the percentage of nucleoside units within the core. In some embodiments, the core is 50% or more of the total oligonucleotide length.

[0131] In some embodiments of oligonucleotides having an asymmetric format, the wings have a length of 6 bases. A non-limiting example of such an oligonucleotide is WV-12485. In some embodiments of oligonucleotides having an asymmetric format, the wings have a length of 7 bases. A non-limiting example of such an oligonucleotide is WV-12107.

[0132] In some embodiments, oligonucleotides having an asymmetric format comprise a wing-core-wing structure, where one wing is different from the other wing. In some embodiments, the wings comprise one or more sugar modifications. In some embodiments, the two wings of the wing-core-wing structure comprise different sugar modifications. In some embodiments, the sugar modifications provide improved stability compared to the absence of the sugar modifications.

[0133] In some embodiments, the core does not include a 2'-substitution. In some embodiments, each sugar unit of the core is a natural sugar unit present in natural, unmodified DNA. In some embodiments, the core includes one or more 2'-halogen modifications. In some embodiments, the core includes one or more 2'-F modifications.

[0134] In some embodiments, some sugar modifications, such as 2'-MOE, increase stability against nucleases. In some embodiments, the wings comprise a 2'-MOE modification. In some embodiments, each nucleoside unit of the wing comprising a pyrimidine base (e.g., C, U, T, etc.) comprises a 2'-MOE modification. In some embodiments, each sugar unit of the wing comprises a 2'-MOE modification. In some embodiments, each nucleoside unit of the wing comprising a purine base (e.g., A, G, etc.) does not comprise a 2'-MOE modification (e.g., 2'-OMe, no 2'-modification, etc.). In some embodiments, each nucleoside unit of the wing comprising a purine base comprises a 2'-OMe modification. In some embodiments, each internucleotide linkage at the 3' position of a sugar unit comprising a 2'-MOE modification is a natural phosphate linkage. In some embodiments, each internucleotide linkage at the 3' position of a sugar unit containing a 2'-MOE modification is a natural phosphate linkage, except that if the wing is a 5' wing relative to the core, the first internucleotide linkage of the wing is a modified internucleotide linkage, e.g., a phosphorothioate diester linkage, and the internucleotide linkage connecting the 3'-terminal nucleoside unit of the wing to the 5'-terminal nucleoside unit of the core is a modified internucleotide linkage, e.g., a phosphorothioate diester linkage; if the wing is a 3' wing relative to the core, the last internucleotide linkage of the wing is a modified internucleotide linkage, e.g., a phosphorothioate diester linkage, and the internucleotide linkage connecting the 3'-terminal nucleoside unit of the core to the 5'-terminal nucleoside unit of the wing is a modified internucleotide linkage, e.g., a phosphorothioate diester linkage. In some embodiments, such a wing is a 5' wing. In some embodiments, such a wing is a 3' wing.

[0135] In some embodiments, the wings do not contain 2'-MOE modifications. In some embodiments, the wings contain 2'-OMe modifications. In some embodiments, each nucleoside unit of the wings independently contains a 2'-OMe modification. Among other things, the present disclosure encompasses the recognition that oligonucleotides containing 2'-OMe modifications are less stable than comparable oligonucleotides containing 2'-MOE modifications under certain conditions. In some embodiments, modified non-natural internucleotide linkages, such as phosphorothioate diester linkages, and in some cases, particularly Sp phosphorothioate diester linkages, can be used to improve the properties, e.g., stability, of the oligonucleotide. In some embodiments, the wings do not contain 2'-MOE modifications, and each internucleotide linkage between the nucleoside units of the wings is a modified internucleotide linkage. In some embodiments, the wings do not contain 2'-MOE modifications, each nucleoside unit of the wings contains a 2'-OMe modification, and each internucleotide linkage between the nucleoside units of the wings is a modified internucleotide linkage. In some embodiments, the modified internucleotide linkage is a phosphorothioate diester linkage. In some embodiments, the modified internucleotide linkage is a chiral-controlled internucleotide linkage. In some embodiments, the modified internucleotide linkage is a chiral-controlled internucleotide linkage, wherein the linking phosphorus is in the Sp conformation. In some embodiments, the modified internucleotide linkage is a chiral-controlled internucleotide linkage, wherein the linking phosphorus is in the Rp conformation. In some embodiments, the modified internucleotide linkage is an Sp phosphorothioate diester linkage. In some embodiments, the modified internucleotide linkage is an Rp phosphorothioate diester linkage. In some embodiments, such a wing is a 5' wing. In some embodiments, such a wing is a 3' wing.

[0136] In some embodiments, 2' modifications and / or modified internucleotide linkages can be used either individually or in combination to fine-tune the properties of the oligonucleotide, such as stability and / or activity.

[0137] In some embodiments, the wings comprise one or more natural phosphate linkages. In some embodiments, the wings comprise one or more consecutive natural phosphate linkages. In some embodiments, the wings comprise one or more natural phosphate linkages and one or more modified internucleotide linkages. In some embodiments, the modified internucleotide linkages are phosphorothioate diester linkages. In some embodiments, the modified internucleotide linkages are Sp phosphorothioate diester linkages.

[0138] In some embodiments, the wings do not contain natural phosphate linkages, and each internucleotide linkage of the wings is independently a modified internucleotide linkage. In some embodiments, the modified internucleotide linkage is chiral and chiral-controlled. In some embodiments, the modified internucleotide linkage is a phosphorothioate diester linkage. In some embodiments, the modified internucleotide linkage is an Sp phosphorothioate diester linkage.

[0139] In some embodiments, for oligonucleotides that comprise or are wing-core-wing structures, the two wings differ in that they comprise different levels and / or types of chemical modifications, backbone chiral center stereochemistry, and / or patterns thereof. In some embodiments, the two wings differ in that they comprise different levels and / or types of sugar modifications, and / or internucleotide linkages, and / or internucleotide linkage stereochemistry, and / or patterns thereof. For example, in some embodiments, one wing comprises a 2'OR modification (R is an optionally substituted C 1~6In addition, or alternatively, one wing may contain natural phosphate linkages while the other wing contains no natural phosphate linkages or a lower level (e.g., number and / or percentage) of natural phosphate linkages; in addition, or alternatively, one wing may contain a particular type of modified internucleotide linkage (e.g., a phosphorothioate diester internucleotide linkage), In some embodiments, one wing contains one or more natural phosphate linkages and one or more 2'-OR modifications (R is not -H or -Me), while the other wing contains no natural phosphate linkages or a lower level (e.g., in number and / or percentage) of that type of modified internucleotide linkage; Additionally or alternatively, one wing may contain chiral modified internucleotide linkages with a bound phosphorus atom in a particular conformation (e.g., Rp or Sp), while the other wing contains no chiral modified internucleotide linkages with a bound phosphorus atom in a particular conformation, or a lower level thereof; Additionally or alternatively, each wing may contain a different pattern of sugar modifications, internucleotide linkages, and / or backbone chiral centers. In some embodiments, one wing contains one or more natural phosphate linkages and one or more 2'-OR modifications (R is not -H or -Me), while the other wing contains no natural phosphate linkages and no 2'-OR modifications (R is not -H or -Me). In some embodiments, one wing comprises one or more native phosphate linkages and one or more 2'-MOE modifications, each internucleotide linkage of the other wing is a phosphorothioate linkage, and each sugar unit of the other wing comprises a 2'-OMe modification. In some embodiments, one wing comprises one or more native phosphate linkages and one or more 2'-MOE modifications, each internucleotide linkage of the other wing is a Sp phosphorothioate linkage, and each sugar unit of the other wing comprises a 2'-OMe modification.

[0140] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where one wing comprises 2'-OMe and the other wing comprises a bicyclic sugar, hi some embodiments, the oligonucleotide comprises a wing-core-wing structure, where one wing comprises 2'-OMe and the other wing comprises a bicyclic sugar, and the majority of the sugars in the core comprise 2'-deoxy.

[0141] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars of one wing comprise 2'-OMe and the majority of sugars of the other wing are bicyclic sugars, hi some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars of one wing comprise 2'-OMe and the majority of sugars of the other wing are bicyclic sugars, and the majority of sugars of the core comprise 2'-deoxy.

[0142] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-OMe, and in the other wing, at least one sugar is a bicyclic sugar and at least one sugar comprises 2'-OMe. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-OMe, and in the other wing, at least one sugar is a bicyclic sugar and at least one sugar comprises 2'-OMe, and the majority of sugars in the core comprise 2'-deoxy.

[0143] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing are bicyclic sugars and in the other wing, at least one sugar is a bicyclic sugar and at least one sugar comprises 2'-OMe. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing are bicyclic sugars and in the other wing, at least one sugar is a bicyclic sugar and at least one sugar comprises 2'-OMe, and the majority of sugars in the core comprise 2'-deoxy.

[0144] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-OMe, and at least two sugars in the other wing are bicyclic sugars, and at least two sugars comprise 2'-OMe. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-OMe, and at least two sugars in the other wing are bicyclic sugars, and at least two sugars comprise 2'-OMe, and the majority of sugars in the core comprise 2'-deoxy.

[0145] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing are bicyclic sugars and in the other wing, at least two sugars are bicyclic sugars and at least two sugars comprise 2'-OMe. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing are bicyclic sugars and in the other wing, at least two sugars are bicyclic sugars and at least two sugars comprise 2'-OMe, and the majority of sugars in the core comprise 2'-deoxy.

[0146] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where each sugar of one wing comprises a 2'-OMe and each sugar of the other wing comprises a bicyclic sugar, hi some embodiments, the oligonucleotide comprises a wing-core-wing structure, where each sugar of one wing comprises a 2'-OMe and each sugar of the other wing comprises a bicyclic sugar, and most sugars of the core comprise 2'-deoxy.

[0147] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, in which each sugar of one wing comprises a bicyclic sugar, each sugar of the other wing comprises a 2'-OMe, and each sugar of the core comprises a 2'-deoxy.

[0148] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where one wing comprises a bicyclic sugar and the other wing comprises a 2'-MOE, hi some embodiments, the oligonucleotide comprises a wing-core-wing structure, where one wing comprises a bicyclic sugar and the other wing comprises a 2'-MOE, and the majority of the sugars in the core comprise 2'-deoxy.

[0149] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars of one wing comprise bicyclic sugars and the majority of sugars of the other wing comprise 2'-MOE. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars of one wing comprise bicyclic sugars and the majority of sugars of the other wing comprise 2'-MOE, and the majority of sugars of the core comprise 2'-deoxy.

[0150] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise bicyclic sugars, and in the other wing, at least one sugar comprises a 2'-MOE and at least one sugar is a bicyclic sugar. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise bicyclic sugars, and in the other wing, at least one sugar comprises a 2'-MOE and at least one sugar is a bicyclic sugar, and the majority of sugars in the core comprise 2'-deoxy.

[0151] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-MOE, at least one sugar in the other wing comprises 2'-MOE, and at least one sugar is a bicyclic sugar. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-MOE, at least one sugar in the other wing comprises 2'-MOE, at least one sugar is a bicyclic sugar, and the majority of sugars in the core comprise 2'-deoxy.

[0152] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise bicyclic sugars and in the other wing, at least two sugars comprise 2'-MOE and at least two sugars are bicyclic sugars. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise bicyclic sugars and in the other wing, at least two sugars comprise 2'-MOE and at least two sugars are bicyclic sugars, and the majority of sugars in the core comprise 2'-deoxy.

[0153] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-MOE, and in the other wing, at least two sugars comprise 2'-MOE, and at least two sugars are bicyclic sugars. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-MOE, and in the other wing, at least two sugars comprise 2'-MOE, and at least two sugars are bicyclic sugars, and the majority of sugars in the core comprise 2'-deoxy.

[0154] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where each sugar of one wing is a bicyclic sugar and each sugar of the other wing comprises a 2'-MOE, hi some embodiments, the oligonucleotide comprises a wing-core-wing structure, where each sugar of one wing is a bicyclic sugar and each sugar of the other wing comprises a 2'-MOE, and most sugars of the core comprise 2'-deoxy.

[0155] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, in which each sugar of one wing comprises a 2'-MOE, each sugar of the other wing is a bicyclic sugar, and each sugar of the core comprises a 2'-deoxy.

[0156] In some embodiments, the bicyclic sugar is LNA, cEt, or BNA.

[0157] In some embodiments, the oligonucleotide comprises a wing-core-wing structure where one wing comprises 2'-OMe and the other wing comprises 2'-F. In some embodiments, the oligonucleotide comprises a wing-core-wing structure where one wing comprises 2'-OMe and the other wing comprises 2'-F, and the majority of the sugars in the core comprise 2'-deoxy.

[0158] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars of one wing comprise 2'-OMe and the majority of sugars of the other wing are 2'-F. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars of one wing comprise 2'-OMe and the majority of sugars of the other wing are 2'-F, and the majority of sugars of the core comprise 2'-deoxy.

[0159] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-OMe, and in the other wing, at least one sugar is 2'-F and at least one sugar comprises 2'-OMe. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-OMe, and in the other wing, at least one sugar is 2'-F and at least one sugar comprises 2'-OMe, and the majority of sugars in the core comprise 2'-deoxy.

[0160] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing are 2'-F and in the other wing, at least one sugar is 2'-F and at least one sugar comprises 2'-OMe. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing are 2'-F and in the other wing, at least one sugar is 2'-F and at least one sugar comprises 2'-OMe, and the majority of sugars in the core comprise 2'-deoxy.

[0161] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-OMe, and in the other wing, at least two sugars are 2'-F and at least two sugars comprise 2'-OMe. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-OMe, and in the other wing, at least two sugars are 2'-F and at least two sugars comprise 2'-OMe, and the majority of sugars in the core comprise 2'-deoxy.

[0162] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing are 2'-F and in the other wing, at least two sugars are 2'-F and at least two sugars comprise 2'-OMe. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing are 2'-F and in the other wing, at least two sugars are 2'-F and at least two sugars comprise 2'-OMe, and the majority of sugars in the core comprise 2'-deoxy.

[0163] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where each sugar of one wing comprises 2'-OMe and each sugar of the other wing comprises 2'-F. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where each sugar of one wing comprises 2'-OMe and each sugar of the other wing comprises 2'-F, and the majority of the sugars of the core comprise 2'-deoxy.

[0164] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, in which each sugar of one wing comprises a 2'-F, each sugar of the other wing comprises a 2'-OMe, and each sugar of the core comprises a 2'-deoxy.

[0165] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where one wing comprises a 2'-F and the other wing comprises a 2'-MOE. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where one wing comprises a 2'-F and the other wing comprises a 2'-MOE, and the core most sugar comprises a 2'-deoxy.

[0166] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars of one wing comprise 2'-F and the majority of sugars of the other wing comprise 2'-MOE. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars of one wing comprise 2'-F and the majority of sugars of the other wing comprise 2'-MOE, and the majority of sugars of the core comprise 2'-deoxy.

[0167] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-F, and in the other wing, at least one sugar comprises 2'-MOE and at least one sugar is 2'-F. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-F, and in the other wing, at least one sugar comprises 2'-MOE and at least one sugar is 2'-F, and the majority of sugars in the core comprise 2'-deoxy.

[0168] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-MOE, and in the other wing, at least one sugar comprises 2'-MOE and at least one sugar is 2'-F. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-MOE, and in the other wing, at least one sugar comprises 2'-MOE and at least one sugar is 2'-F, and the majority of sugars in the core comprise 2'-deoxy.

[0169] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-F, and in the other wing, at least two sugars comprise 2'-MOE and at least two sugars are 2'-F. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-F, and in the other wing, at least two sugars comprise 2'-MOE and at least two sugars are 2'-F, and the majority of sugars in the core comprise 2'-deoxy.

[0170] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-MOE, and in the other wing, at least two sugars comprise 2'-MOE and at least two sugars are 2'-F. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where the majority of sugars in one wing comprise 2'-MOE, and in the other wing, at least two sugars comprise 2'-MOE and at least two sugars are 2'-F, and the majority of sugars in the core comprise 2'-deoxy.

[0171] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where each sugar of one wing is 2'-F and each sugar of the other wing comprises 2'-MOE. In some embodiments, the oligonucleotide comprises a wing-core-wing structure, where each sugar of one wing is 2'-F and each sugar of the other wing comprises 2'-MOE, and the majority of the sugars of the core comprise 2'-deoxy.

[0172] In some embodiments, the oligonucleotide comprises a wing-core-wing structure, in which each sugar of one wing comprises 2'-MOE, each sugar of the other wing is 2'-F, and each sugar of the core comprises 2'-deoxy.

[0173] In some embodiments, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% or more of the sugar moieties of a provided oligonucleotide are modified. In some embodiments, each sugar moiety of a provided oligonucleotide is modified. In some embodiments, a modified sugar moiety comprises a 2'-modification. In some embodiments, a modified sugar moiety comprises a 2'-modification. In some embodiments, a 2'-modification is a 2'-OR 1 In some embodiments, the 2'-modification is 2'-OMe. In some embodiments, the 2'-modification is 2'-MOE. In some embodiments, the 2'-modification is an LNA sugar modification. In some embodiments, the 2'-modification is 2'-F. In some embodiments, each sugar modification is independently a 2'-modification. In some embodiments, each sugar modification is independently a 2'-OR 1 or 2'-F. In some embodiments, each sugar modification is independently 2'-OR 1 or 2'-F, wherein R 1 is an optionally substituted C 1~6 In some embodiments, each sugar modification is independently 2'-OR alkyl. 1 or 2'-F, where at least one is 2'-F. In some embodiments, each sugar modification is independently 2'-OR 1 or 2'-F, wherein R 1is an optionally substituted C 1~6 alkyl, in which case at least one is 2'-OR 1 In some embodiments, each sugar modification is independently 2'-OR 1 or 2'-F, in which case at least one is 2'-F and at least one is 2'-OR 1 In some embodiments, each sugar modification is independently 2'-OR 1 or 2'-F, wherein R 1 is an optionally substituted C 1~6 alkyl, where at least one is 2'-F and at least one is 2'-OR 1 is.

[0174] In some embodiments, a nucleoside comprising a 2'-modification is followed by a modified internucleotide linkage. In some embodiments, a nucleoside comprising a 2'-modification is preceded by a modified internucleotide linkage. In some embodiments, the modified internucleotide linkage is a chiral internucleotide linkage. In some embodiments, the modified internucleotide linkage is a phosphorothioate. In some embodiments, the chiral internucleotide linkage is Sp. In some embodiments, a nucleoside comprising a 2'-modification is followed by an Sp chiral internucleotide linkage. In some embodiments, a nucleoside comprising a 2'-F is followed by an Sp chiral internucleotide linkage. In some embodiments, a nucleoside comprising a 2'-modification is preceded by an Sp chiral internucleotide linkage. In some embodiments, a nucleoside comprising a 2'-F is preceded by an Sp chiral internucleotide linkage. In some embodiments, the chiral internucleotide linkage is Rp. In some embodiments, a nucleoside comprising a 2'-modification is followed by an Rp chiral internucleotide linkage. In some embodiments, a nucleoside containing a 2'-F is followed by an Rp chiral internucleotide linkage. In some embodiments, a nucleoside containing a 2'-modification is preceded by an Rp chiral internucleotide linkage. In some embodiments, a nucleoside containing a 2'-F is preceded by an Rp chiral internucleotide linkage.

[0175] In some embodiments, the provided oligonucleotides have an asymmetric format and have a wing-core-wing structure. In some embodiments of oligonucleotides having an asymmetric format, one wing differs from the other wing. In some embodiments of oligonucleotides having an asymmetric format, one wing differs from the other wing in a sugar modification, or a combination or pattern thereof, or a backbone internucleotide linkage, or a combination or pattern thereof, or a backbone chiral center, or a combination or pattern thereof. In some embodiments of oligonucleotides having an asymmetric format, the core comprises one or more 2'-deoxy sugars. In some embodiments of oligonucleotides having an asymmetric format, the core comprises five or more consecutive 2'-deoxy sugars. In some embodiments of oligonucleotides having an asymmetric format, the core comprises 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or more consecutive 2'-deoxy sugars. Some non-limiting examples of oligonucleotides having an asymmetric format are provided herein. In some embodiments of oligonucleotides having an asymmetric format, the first wing and the second wing independently have a pattern of sugar 2'-modifications that is or includes the following: F, FF, FFF, FFFF, FFFFF, FFFFF, FMMMF, FMMMF, LMMMm, m, M, mm, MM, mmm, mmMm, MMm, MMM, mmm, mmmm, mmMMm, MMMm, MMMM, mmmm, mmmmm, MMMMM, mmMMm, MMMMM, mmmmm, or any pattern of 2'-modifications of any wing of any oligonucleotide described herein, where the patterns of 2'-modification of the first and second wing are different and m=2'-OMe; M=2'-MOE; F=2'-F; and L=LNA.

[0176] In some embodiments, oligonucleotides having an asymmetric format (e.g., one wing chemically different from the other wing) have improved biological activity compared to oligonucleotides having the same base sequence but a different structure (e.g., a symmetric format in which both wings have the same pattern of chemical modifications; or a different asymmetric format). In some embodiments, the improved biological activity includes improved expression, activity, and / or level or reduction of a gene or gene product. In some embodiments, the improved biological activity is improved delivery to a cell nucleus. In some embodiments, the improved biological activity is improved delivery to a cell nucleus where one wing in the oligonucleotide having an asymmetric format comprises one 2'-F or two or more 2'-F. In some embodiments, the improved biological activity is improved delivery to a cell nucleus where one wing in the oligonucleotide having an asymmetric format comprises one 2'-MOE or two or more 2'-MOE. In some embodiments, the improved biological activity is improved delivery to a cell nucleus where one wing in the oligonucleotide having an asymmetric format comprises one 2'-OMe or two or more 2'-OMe. In some embodiments, the improved biological activity is improved delivery to the cell nucleus, and one wing in the oligonucleotide having an asymmetric format comprises one bicyclic sugar or two or more bicyclic sugars.

[0177] In some embodiments, oligonucleotides having an asymmetric format comprise a first wing having a particular sugar modification or combination or pattern thereof and a second wing having a different particular sugar modification or combination or pattern thereof.

[0178] In some embodiments, oligonucleotides having an asymmetric format comprise a first wing having a particular 2'-sugar modification or combination or pattern thereof and a second wing having a different particular 2'-sugar modification or combination or pattern thereof.

[0179] In some embodiments, the pattern of sugar modifications of oligonucleotides having an asymmetric format is as follows: S1-S1-S1-S1, S1-S1-S1-S2, S1-S1-S2-S1, S1-S1-S2-S2, S1-S2-S1-S1, S1-S2-S1-S2, S1-S2-S1-S2, S1-S2-S2-S1, S1-S2-S2-S2, S2-S1-S1-S1, S2-S1-S1-S2, S2-S1-S1-S2, S2-S1-S2-S1, S2-S1-S2-S2, S2-S2-S1-S1, S2-S2-S1-S2, S2-S2-S2-S1, S2-S2-S2-S2, S1-S1-S1-S1, S1-S1-S1-S3, S1-S1-S3-S1, S1-S1-S3-S3, S1-S3-S1-S1, S1-S3-S1-S3, S1-S3-S3-S1, S1-S3-S3-S3, S3-S1-S1-S1, S3-S1-S1-S3, S3-S1-S1-S3, S3-S1-S3-S1, S3-S1-S3-S1, S3-S1-S3-S3, S3-S1-S3-S3, S3-S3-S1-S1, S3-S3-S1-S1, S3-S3-S1-S3, S3-S3-S1-S3, S3-S3-S3-S1, S3-S3-S3-S1, S3-S3-S3-S3, S3-S3-S3-S3, S1-S1-S1-S1, S1-S1-S1-S4, S1-S1-S1-S4, S1-S1-S4-S1, S1-S1-S4-S1, S1-S1-S4-S4, S1-S1-S4-S4, S1-S4-S1-S1, S1-S4-S1-S1, S1-S4-S1-S4, S1-S4-S1-S4, S1-S4-S4-S1, S1-S4-S4-S1, S1-S4-S4-S4, S1-S4-S4-S4, S4-S1-S1-S1, S4-S1-S1-S1, S4-S1-S1-S4, S4-S1-S1-S4, S4-S1-S4-S1, S4-S1-S4-S1, S4-S1-S4-S4, S4-S1-S4-S4, S4-S4-S1-S1, S4-S4-S1-S1, S4-S4-S1-S4, S4-S4-S1-S4, S4-S4-S4-S1, S4-S4-S4-S1, S4-S4-S4-S4, S4-S4-S4-S4, S1-S1-S1-S2, S1-S1-S1-S3, S1-S1-S1-S3, S1-S1-S1-S3, S1-S1-S2-S1, S1-S1-S2-S1, S1-S1-S2-S1,S1-S1-S2-S2、S1-S1-S2-S2、S1-S1-S2-S2、S1-S1-S2-S3、S1-S1-S2-S3、S1 -S1-S2-S3、S1-S1-S3-S1、S1-S1-S3-S1、S1-S1-S3-S1、S1-S1-S3-S2、S1-S 1-S3-S2, S1-S1-S3-S2, S1-S1-S3-S3, S1-S1-S3-S3, S1-S2-S1-S1, S1-S2-S1-S1, S1-S2-S1-S1, S1-S2-S1-S1, S1-S2-S1-S2, S1-S2-S1-S2, S1-S2-S1-S2 S2, S1-S2-S1-S3, S1-S2-S1-S3, S1-S2-S1-S3, S1-S2-S2-S1, S1-S2-S2-S1, S1-S2-S2-S1, S1-S2-S2-S2, S1-S2-S2-S2, S1-S2-S2-S2, S1-S2-S2-S3, S 1-S2-S2-S3、S1-S2-S2-S3、S1-S2-S3-S1、S1-S2-S3-S1、S1-S2-S3-S1、S1-S2-S3-S2、S1-S2-S3-S2、S1-S2-S3-S3、S1-S2-S3-S3、S1-S2- S3-S3、S1-S3-S1-S1、S1-S3-S1-S1、S1-S3-S1-S1、S1-S3-S1-S2、S1-S3-S1 -S2、S1-S3-S1-S2、S1-S3-S1-S3、S1-S3-S1-S3、S1-S3-S1-S3、S1-S3-S2-S 1. S1-S3-S2-S1 S3-S3-S1, S1-S3-S3-S2, S1-S3-S3-S2, S1-S3-S3-S2, S1-S3-S3-S3, S1-S3-S3-S3, S1-S3-S3-S3, S2-S1-S1-S1, S2-S1-S1-S1, S2-S1-S1-S1, S2-S1-S 1-S2, S2-S1-S1-S2, S2-S1-S1-S2, S2-S1-S1-S3, S2-S1-S1-S3, S2-S1-S1-S3, S2-S1-S2-S1, S2-S1-S2-S1, S2-S1-S2-S1, S2-S1-S2-S2, S2-S1-S2-S2S2-S1-S2-S2, S2-S1-S2-S3, S2-S1-S2-S3, S2-S1-S2-S3, S2-S1-S3-S1, S2-S1-S3-S1, S2-S1-S3-S1, S2-S1-S3-S2, S2-S1-S3-S2, S2-S1-S3-S2, S2-S 1-S3-S3, S2-S1-S3-S3, S2-S1-S3-S3, S2-S2-S1-S1, S2-S2-S1-S1, S2-S2-S1-S1, S2-S2-S1-S2, S2-S2-S1-S2, S2-S2-S1-S2, S2-S2-S1-S3, S2-S2-S1- S3, S2-S2-S1-S3, S2-S2-S2-S1, S2-S2-S2-S1, S2-S2-S2-S1, S2-S2-S2-S2, S2-S2-S2-S2, S2-S2-S2-S2, S2-S2-S2-S3, S2-S2-S2-S3, S 2-S2-S3-S1、S2-S2-S3-S1、S2-S2-S3-S1、S2-S2-S3-S2、S2-S2-S3-S2、S2-S2-S3-S2、S2-S2-S3-S3、S2-S2-S3-S3、S2-S3-S1-S1、S2-S3- S1-S1, S2-S3-S1-S1, S2-S3-S1-S2, S2-S3-S1-S2, S2-S3-S1-S2, S2-S3-S1-S3, S2-S3-S1-S3, S2-S3-S1-S3, S2-S3-S2-S1, S2-S3-S2-S1, S2-S3-S2-S 1. S2-S3-S2-S2, S2-S3-S2-S2, S2-S3-S2-S2, S2-S3-S2-S3, S2-S3-S2-S3, S2-S3-S2-S3, S2-S3-S3-S1, S2-S3-S3-S1, S2-S3-S3-S1, S2-S3-S3-S2, S2- S3-S3-S2, S2-S3-S3-S2, S2-S3-S3-S3, S2-S3-S3-S3, S2-S3-S3-S3, S3-S1-S1-S1, S3-S1-S1-S1, S3-S1-S1-S1, S3-S1-S1-S2, S3-S1-S1-S2, S3-S1-S 1-S2, S3-S1-S1-S3, S3-S1-S1-S3, S3-S1-S1-S3, S3-S1-S2-S1, S3-S1-S2-S1, S3-S1-S2-S1, S3-S1-S2-S2, S3-S1-S2-S2, S3-S1-S2-S2, S3-S1-S2-S3S3-S1-S2-S3, S3-S1-S2-S3, S3-S1-S3-S1, S3-S1-S3-S1, S3-S1-S3-S1, S3-S1-S3-S2, S3-S1-S3-S2, S3-S1-S3-S2, S3-S1-S3-S3, S3-S1-S3-S3, S3-S 1-S3-S3、S3-S2-S1-S1、S3-S2-S1-S1、S3-S2-S1-S1、S3-S2-S1-S2、S3-S2-S1-S2、S3-S2-S1-S2、S3-S2-S1-S3、S3-S2-S1-S3、S3-S2-S2- S1, S3-S2-S2-S1, S3-S2-S2-S1, S3-S2-S2-S2, S3-S2-S2-S2, S3-S2-S2-S2, S3-S2-S2-S3, S3-S2-S2-S3, S3-S2-S2-S3, S3-S2-S3-S1, S3-S2-S3-S1, S 3-S2-S3-S1、S3-S2-S3-S2、S3-S2-S3-S2、S3-S2-S3-S2、S3-S2-S3-S3、S3-S2-S3-S3、S3-S2-S3-S3、S3-S3-S1-S1、S3-S3-S1-S1、S3-S3-S1-S1、S3-S3- S1-S2, S3-S3-S1-S2, S3-S3-S1-S2, S3-S3-S1-S3, S3-S3-S1-S3, S3-S3-S1-S3, S3-S3-S2-S1, S3-S3-S2-S1, S3-S3-S2-S1, S3-S3-S2-S2, S3-S3-S2-S 2, S3-S3-S2-S2, S3-S3-S2-S3, S3-S3-S2-S3, S3-S3-S2-S3, S3-S3-S3-S1, S3-S3-S3-S1, S3-S3-S3-S1, S3-S3-S3-S2, S3-S3-S3-S2, S3-S3-S3-S2, S3- S3-S3-S3、S3-S3-S3-S3、S3-S3-S3-S3、S3-S3-S3-S3、S1-S1-S1-S2、S1-S1 -S1-S4、S1-S1-S1-S4、S1-S1-S1-S4、S1-S1-S2-S1、S1-S1-S2-S1、S1-S1-S 2-S1、S1-S1-S2-S2、S1-S1-S2-S2、S1-S1-S2-S2、S1-S1-S2-S4、S1-S1-S2- S4、S1-S1-S2-S4、S1-S1-S4-S1、S1-S1-S4-S1、S1-S1-S4-S1、S1-S1-S4-S2、S1-S1-S4-S2、S1-S1-S4-S2、S1-S1-S4-S4、S1-S1-S4-S4、S1-S1-S4-S4、S1-S2-S1-S1、S1-S2-S1-S1、S1-S2-S1-S1、S1-S2-S1-S2、S1-S2-S1-S2、S1-S2-S1-S2、S1-S2-S1-S4、S1-S2-S1-S4、S1-S2-S1-S4、S1-S2-S2-S1、S1-S2-S2-S1、S1-S2-S2-S1、S1-S2-S2-S2、S1-S2-S2-S2、S1-S2-S2-S2、S1-S2-S2-S4、S1-S2-S2-S4、S1-S2-S2-S4、S1-S2-S4-S1、S1-S2-S4-S1、S1-S2-S4-S1、S1-S2-S4-S2、S1-S2-S4-S2、S1-S2-S4-S2、S1-S2-S4-S4、S1-S2-S4-S4、S1-S2-S4-S4、S1-S4-S1-S1、S1-S4-S1-S1、S1-S4-S1-S1、S1-S4-S1-S2、S1-S4-S1-S2、S1-S4-S1-S2、S1-S4-S1-S4、S1-S4-S1-S4、S1-S4-S1-S4、S1-S4-S2-S1、S1-S4-S2-S1、S1-S4-S2-S1、S1-S4-S2-S2、S1-S4-S2-S2、S1-S4-S2-S2、S1-S4-S2-S4、S1-S4-S2-S4、S1-S4-S2-S4、S1-S4-S4-S1、S1-S4-S4-S1、S1-S4-S4-S1、S1-S4-S4-S2、S1-S4-S4-S2、S1-S4-S4-S2、S1-S4-S4-S4、S1-S4-S4-S4、S1-S4-S4-S4、S2-S1-S1-S1、S2-S1-S1-S1、S2-S1-S1-S1、S2-S1-S1-S2、S2-S1-S1-S2、S2-S1-S1-S2、S2-S1-S1-S4、S2-S1-S1-S4、S2-S1-S1-S4、S2-S1-S2-S1、S2-S1-S2-S1、S2-S1-S2-S1、S2-S1-S2-S2、S2-S1-S2-S2、S2-S1-S2-S2、S2-S1-S2-S4、S2-S1-S2-S4、S2-S1-S2-S4、S2-S1-S4-S1、S2-S1-S4-S1、S2-S1-S4-S1、S2-S1-S4-S2、S2-S1-S4-S2、S2-S1-S4-S2、S2-S1-S4-S4, S2-S1-S4-S4, S2-S1-S4-S4, S2-S2-S1-S1, S2-S2-S1-S1, S2-S2-S1-S1, S2-S2-S1-S2, S2-S2-S1-S2, S2-S2-S1-S2, S2-S2-S1-S4, S2-S2-S1-S4, S2-S2-S1-S 4、S2-S2-S2-S1、S2-S2-S2-S1、S2-S2-S2-S1、S2-S2-S2-S2、S2-S2-S2-S2、S2-S2-S2-S2、S2-S2-S2-S4、S2-S2-S2-S4、S2-S2-S2-S4、S2-S2-S4-S1、S2-S2-S4-S1、S2-S2-S4-S1、S2-S2-S4-S2、S2-S2-S4-S2、S2-S2-S4-S2、S2-S2-S4-S4、S2-S2-S4-S4、S2-S2-S4-S4、S2-S4-S1-S1、S2-S4-S1-S1、S2-S4-S1-S1、S2-S4-S1-S2、S2-S4-S1-S2、S2-S4-S1-S2、S2-S4-S1-S4、S2-S4-S1-S4、S2-S4-S1-S4、S2-S4-S2-S1、S2-S4-S2-S1、S2-S4-S2-S1、S2-S4-S2-S2、S2-S4-S2-S2、S2-S4-S2-S2、S2-S4-S2-S4、S2-S4-S2-S4、S2-S4-S2-S4、S2-S4-S4-S1、S2-S4-S4-S1、S2-S4-S4-S1、S2-S4-S4-S2、S2-S4-S4-S2、S2-S4-S4-S2、S2-S4-S4-S4、S2-S4-S4-S4、S2-S4-S4-S4、S4-S1-S1-S1、S4-S1-S1-S1、S4-S1-S1-S1、S4-S1-S1-S2、S4-S1-S1-S2、S4-S1-S1-S2、S4-S1-S1-S4、S4-S1-S1-S4、S4-S1-S1-S4、S4-S1-S2-S1、S4-S1-S2-S1、S4-S1-S2-S1、S4-S1-S2-S2、S4-S1-S2-S2、S4-S1-S2-S2、S4-S1-S2-S4、S4-S1-S2-S4、S4-S1-S2-S4、S4-S1-S4-S1、S4-S1-S4-S1、S4-S1-S4-S1、S4-S1-S4-S2、S4-S1-S4-S2、S4-S1-S4-S2、S4-S1-S4-S4、S4-S1-S4-S4、S4-S1-S4-S4、S4-S2-S1-S1、S4-S2-S1-S1、S4-S2-S1-S1、S4-S2-S1-S2、S4-S2-S1-S2、S4-S2-S1-S2、S4-S2-S1-S4、S4-S2-S1-S4、S4-S2-S1-S4、S4-S2-S2-S1、S4-S2-S2-S1、S4-S2-S2-S1, S4-S2-S2-S2, S4-S2-S2-S2, S4-S2-S2-S2, S4-S2-S2-S4, S4-S2-S2-S4 , S4-S2-S2-S4, S4-S2-S4-S1, S4-S2-S4-S1, S4-S2-S4-S1, S4-S2-S4-S2, S4-S2-S4-S2 , S4-S2-S4-S2, S4-S2-S4-S4, S4-S2-S4-S4, S4-S2-S4-S4, S4-S4-S1-S1, S4-S4-S1-S1 , S4-S4-S1-S1, S4-S4-S1-S2, S4-S4-S1-S2, S4-S4-S1-S2, S4-S4-S1-S4, S4-S4-S1-S4 , S4-S4-S1-S4, S4-S4-S2-S1, S4-S4-S2-S1, S4-S4-S2-S1, S4-S4-S2-S2, S4-S4-S2-S 2, S4-S4-S2-S2, S4-S4-S2-S4, S4-S4-S2-S4, S4-S4-S2-S4, S4-S4-S4-S1, S4-S4-S4-S and S4-S4-S4-S1, S4-S4-S4-S2, S4-S4-S4-S2, S4-S4-S4-S2, S4-S4-S4-S4, S4-S4-S4-S4, S4-S4-S4-S4, or S4-S4-S4-S4, where S1, S2, S3, and S4 are different types of sugar modifications. In some embodiments, S1, S2, S3, and S4 are different types of 2'-sugar modifications. In some embodiments, such sugar modification patterns are present in the first wing, second wing, and / or core of an oligonucleotide having an asymmetric format.

[0180] In some embodiments, the pattern of sugar modifications of oligonucleotides having an asymmetric format is as follows: S1-S1-S1-S1-S1, S1-S1-S1-S1-S2, S1-S1-S1-S2-S1, S1-S1-S1-S2-S2, S1-S1-S2-S1-S1, S1-S1-S2-S1-S2, S1-S1-S2-S2-S1, S1-S1-S2-S2-S2, S1-S2-S1-S1-S1, S1-S2-S1-S1-S2, S1-S2-S1-S2-S1, S1-S2-S1-S2-S2, S1-S2-S2-S1-S1, S1-S2-S2-S1-S2, S1-S2-S2-S2-S1, S1-S2-S2-S2-S2, S2-S1-S1-S1-S1, S2-S1-S1-S1-S2, S2-S1-S1-S2-S1, S2-S1-S1-S2-S2, S2-S1-S2-S1-S1, S2-S1-S2-S1-S2, S2-S1-S2-S2-S1, S2-S1-S2-S2-S2, S2-S2-S1-S1-S1, S2-S2-S1-S1-S2, S2-S2-S1-S2-S1, S2-S2-S1-S2-S2, S2-S2-S2-S1-S1, S2-S2-S2-S1-S2, S2-S2-S2-S2-S1, S2-S2-S2-S2-S2, S1-S1-S1-S1-S1, S1-S1-S1-S1-S3, S1-S1-S1-S3-S1, S1-S1-S1-S3-S3, S1-S1-S3-S1-S1, S1-S1-S3-S1-S3, S1-S1-S3-S3-S1, S1-S1-S3-S3-S3, S1-S3-S1-S1-S1, S1-S3-S1-S1-S3, S1-S3-S1-S3-S1, S1-S3-S1-S3-S3, S1-S3-S3-S1-S1, S1-S3-S3-S1-S3, S1-S3-S3-S3-S1, S1-S3-S3-S3-S3, S3-S1-S1-S1-S1, S3-S1-S1-S1-S3, S3-S1-S1-S3-S1, S3-S1-S1-S3-S3, S3-S1-S3-S1-S1, S3-S1-S3-S1-S3, S3-S1-S3-S3-S1, S3-S1-S3-S3-S3, S3-S3-S1-S1-S1, S3-S3-S1-S1-S3, S3-S3-S1-S3-S1, S3-S3-S1-S3-S3, S3-S3-S3-S1-S1, S3-S3-S3-S1-S3, S3-S3-S3-S3-S1,S3-S3-S3-S3-S3-S3、S1-S1-S1-S4、S1-S1-S1-S4-S1、S1-S1-S1-S4-S4、S1-S1-S4-S1-S1、S1-S1-S4-S1-S4、S1-S1-S4-S1-S4、S1-S1-S4-S1-S4、S1-S1-S4-S4-S1、S1-S1-S4-S4-S1、S1- S4-S1-S1-S1、S1-S4-S1-S1-S4、S1-S4-S1-S4-S1、S1-S4-S1-S4-S4、S1-S4-S1-S4-S4、S1-S4-S4-S1-S1、S1-S4-S4-S1-S4、S1-S4-S4-S4-S1、S1-S4-S4-S4-S1、S1-S4-S4-S4-S4、S4-S1-S 1-S1-S1、S4-S1-S1-S1-S4、S4-S1-S1-S4-S1、S4-S1-S1-S4-S4、S4-S1-S4-S1-S1、S4-S1-S4-S1-S1、S4-S1-S4-S1-S4、S4-S1-S4-S4-S1、S4-S1-S4-S4-S1、S4-S1-S4-S4-S1 -S1、S4-S4-S1-S1-S4、S4-S4-S1-S4-S1、S4-S4-S1-S4-S4、S4-S4-S4-S1-S1、S4-S4-S4-S1-S1、S4-S4-S4-S1-S4、S4-S4-S4-S1-S4、S4-S4-S4-S1、S4-S4-S4-S4-S1、S4-S4-S4-S4-S4、S1-S1-S1-S2-S3、 S1-S1-S1-S3-S1、S1-S1-S1-S3-S2、S1-S1-S1-S3-S3、S1-S1-S2-S1-S1、S 1-S1-S2-S1-S2、S1-S1-S2-S1-S3、S1-S1-S2-S2-S1、S1-S1-S2-S2-S2、S1- S1-S2-S2-S3、S1-S1-S2-S3-S1、S1-S1-S2-S3-S2、S1-S1-S2-S3-S3、S1-S1 -S3-S1-S1、S1-S1-S3-S1-S2、S1-S1-S3-S1-S3、S1-S1-S3-S2-S1、S1-S1-S 3-S2-S2、S1-S1-S3-S2-S3、S1-S1-S3-S3-S1、S1-S1-S3-S3-S2、S1-S1-S3- S3-S3、S1-S2-S1-S1-S1、S1-S2-S1-S1-S2、S1-S2-S1-S1-S3、S1-S2-S1-S2 -S1、S1-S2-S1-S2-S2、S1-S2-S1-S2-S3、S1-S2-S1-S3-S1、S1-S2-S1-S3-S 2、S1-S2-S1-S3-S3、S1-S2-S2-S1-S1、S1-S2-S2-S1-S2、S1-S2-S2-S1-S3、S1-S2-S2-S1, S1-S2-S2-S2-S2, S1-S2-S2-S3, S1-S2-S2-S3-S1, S1-S2-S2-S3-S2, S1-S2-S2-S3-S3, S1-S2-S3-S1-S1, S1-S2-S3-S1-S2, S1- S2-S3-S1-S3, S1-S2-S3-S2-S1, S1-S2-S3-S2-S2, S1-S2-S3-S2-S3, S1-S2-S3-S3-S1, S1-S2-S3-S3-S2, S1-S2-S3-S3-S3, S1-S3-S1-S1-S1, S1-S3-S 1-S1-S2, S1-S3-S1-S1-S3, S1-S3-S1-S2-S1, S1-S3-S1-S2-S2, S1-S3-S1-S2-S3, S1-S3-S1-S3-S1, S1-S3-S1-S3-S2, S1-S3-S1-S3-S3, S1-S3-S2-S1 -S1, S1-S3-S2-S1-S2, S1-S3-S2-S1-S3, S1-S3-S2-S2-S1, S1-S3-S2-S2-S2, S1-S3-S2-S2-S3, S1-S3-S2-S3-S1, S1-S3-S2-S3-S2, S1-S3-S2-S3-S3 S1-S3-S3-S1-S1, S1-S3-S3-S1-S2, S1-S3-S3-S1-S3, S1-S3-S3-S2-S1, S1-S3-S3-S2-S2, S1-S3-S3-S2-S3, S1-S3-S3-S3-S1, S1-S3-S3-S3-S2, S1- S3-S3-S3-S3, S2-S1-S1-S1-S1, S2-S1-S1-S1-S2, S2-S1-S1-S1-S3, S2-S1-S1-S2-S1, S2-S1-S1-S2-S2, S2-S1-S1-S2-S3, S2-S1-S1-S3-S1, S2-S1-S 1-S3-S2, S2-S1-S1-S3-S3, S2-S1-S2-S1-S1, S2-S1-S2-S1-S2, S2-S1-S2-S1-S3, S2-S1-S2-S2-S1, S2-S1-S2-S2-S2, S2-S1-S2-S2-S3, S2-S1-S2-S3 -S1, S2-S1-S2-S3-S2, S2-S1-S2-S3-S3, S2-S1-S3-S1-S1, S2-S1-S3-S1-S2, S2-S1-S3-S1-S3, S2-S1-S3-S2-S1, S2-S1-S3-S2-S2, S2-S1-S3-S2-S3S2-S1-S3-S3-S1, S2-S1-S3-S3-S2, S2-S1-S3-S3-S3, S2-S2-S1-S1-S1, S2-S2-S1-S1-S2, S2-S2-S1-S1-S3, S2-S2-S1-S2-S1, S2-S2-S1-S2-S2, S2- S2-S1-S2-S3, S2-S2-S1-S3-S1, S2-S2-S1-S3-S2, S2-S2-S1-S3-S3, S2-S2-S2-S1-S1, S2-S2-S2-S1-S2, S2-S2-S2-S1-S3, S2-S2-S2-S2-S1, S2-S2-S 2-S2-S2, S2-S2-S2-S3, S2-S2-S2-S3-S1, S2-S2-S2-S3-S2, S2-S2-S2-S3-S3, S2-S2-S3-S1-S1, S2-S2-S3-S1-S2, S2-S2-S3-S1-S3, S2-S2-S3-S2 -S1, S2-S2-S3-S2-S2, S2-S2-S3-S2-S3, S2-S2-S3-S3-S1, S2-S2-S3-S3-S2, S2-S2-S3-S3-S3, S2-S3-S1-S1-S1, S2-S3-S1-S1-S2, S2-S3-S1-S1-S3 S2-S3-S1-S2-S1, S2-S3-S1-S2-S2, S2-S3-S1-S2-S3, S2-S3-S1-S3-S1, S2-S3-S1-S3-S2, S2-S3-S1-S3-S3, S2-S3-S2-S1-S1, S2-S3-S2-S1-S2, S2- S3-S2-S1-S3, S2-S3-S2-S2-S1, S2-S3-S2-S2-S2, S2-S3-S2-S2-S3, S2-S3-S2-S3-S1, S2-S3-S2-S3-S2, S2-S3-S2-S3-S3, S2-S3-S3-S1-S1, S2-S3-S 3-S1-S2、S2-S3-S3-S1-S3、S2-S3-S3-S2-S1、S2-S3-S3-S2-S2、S2-S3-S3-S2-S3、S2-S3-S3-S3-S1、S2-S3-S3-S3-S2、S2-S3-S3-S3-S3、S3-S1-S1-S1 -S1, S3-S1-S1-S1-S2, S3-S1-S1-S1-S3, S3-S1-S1-S2-S1, S3-S1-S1-S2-S2, S3-S1-S1-S2-S3, S3-S1-S1-S3-S1, S3-S1-S1-S3-S2, S3-S1-S1-S3-S3S3-S1-S2-S1-S1, S3-S1-S2-S1-S2, S3-S1-S2-S1-S3, S3-S1-S2-S2-S1, S3-S1-S2-S2-S2, S3-S1-S2-S2-S3, S3-S1-S2-S3-S1, S3-S1-S2-S3-S2, S3- S1-S2-S3-S3, S3-S1-S3-S1-S1, S3-S1-S3-S1-S2, S3-S1-S3-S1-S3, S3-S1-S3-S2-S1, S3-S1-S3-S2-S2, S3-S1-S3-S2-S3, S3-S1-S3-S3-S1, S3-S1-S 3-S3-S2, S3-S1-S3-S3-S3, S3-S2-S1-S1-S1, S3-S2-S1-S1-S2, S3-S2-S1-S1-S3, S3-S2-S1-S2-S1, S3-S2-S1-S2-S2, S3-S2-S1-S2-S3, S3-S2-S1-S3 -S1, S3-S2-S1-S3-S2, S3-S2-S1-S3-S3, S3-S2-S2-S1-S1, S3-S2-S2-S1-S2, S3-S2-S2-S1-S3, S3-S2-S2-S2-S1, S3-S2-S2-S2-S2, S3-S2-S2-S2-S3 S3-S2-S2-S3-S1, S3-S2-S2-S3-S2, S3-S2-S2-S3-S3, S3-S2-S3-S1-S1, S3-S2-S3-S1-S2, S3-S2-S3-S1-S3, S3-S2-S3-S2-S1, S3-S2-S3-S2-S2, S3- S2-S3-S2-S3, S3-S2-S3-S3-S1, S3-S2-S3-S3-S2, S3-S2-S3-S3-S3, S3-S3-S1-S1-S1, S3-S3-S1-S1-S2, S3-S3-S1-S1-S3, S3-S3-S1-S2-S1, S3-S3-S 1-S2-S2, S3-S3-S1-S2-S3, S3-S3-S1-S3-S1, S3-S3-S1-S3-S2, S3-S3-S1-S3-S3, S3-S3-S2-S1-S1, S3-S3-S2-S1-S2, S3-S3-S2-S1-S3, S3-S3-S2-S2 -S1, S3-S3-S2-S2-S2, S3-S3-S2-S2-S3, S3-S3-S2-S3-S1, S3-S3-S2-S3-S2, S3-S3-S2-S3-S3, S3-S3-S3-S1-S1, S3-S3-S3-S1-S2, S3-S3-S3-S1-S3S3-S3-S3-S2-S1, S3-S3-S3-S2-S2, S3-S3-S3-S2-S3, S3-S3-S3-S3-S1, S3-S3-S3-S3-S2, S3-S3-S3-S3-S3, S3-S3-S3-S3-S3, S1-S1-S1-S2-S4, S1-S1-S1-S4-S1, S1-S1-S1-S4-S2, S1-S1-S1-S4-S4, S1-S 1-S2-S1-S1、S1-S1-S2-S1-S2、S1-S1-S2-S1-S4、S1-S1-S2-S2-S1、S1-S1-S2-S2-S2、S1-S1-S2-S2-S4、S1-S1-S2-S4-S1、S1-S1-S2-S4-S2、S1-S1-S2-S4-S4、S1-S1-S4-S1-S1、S1-S1-S4-S1-S2、S1-S1-S4-S1-S4、S1-S1-S4-S2-S1、S1-S1-S4-S2-S2、S1-S1-S4-S2-S4、S1-S1-S4-S4-S1、S1-S1-S4-S4-S2、S1-S1-S4-S4-S4、S1-S2-S1-S1-S1、S1-S2-S1-S1-S2、S1-S2-S1-S1-S4、S1-S2-S1-S2-S1、S1-S2-S1-S2-S2、S1-S2-S1-S2-S4、S1-S2-S1-S4-S1、S1-S2-S1-S4-S2、S1-S2-S1-S4-S4、S1-S2-S2-S1-S1、S1-S2-S2-S1-S2、S1-S2-S2-S1-S4、S1-S2-S2-S2-S1、S1-S2-S2-S2-S2、S1-S2-S2-S2-S4、S1-S2-S2-S4-S1、S1-S2-S2-S4-S2、S1-S2-S2-S4-S4、S1-S2-S4-S1-S1、S1-S2-S4-S1-S2、S1-S2-S4-S1-S4、S1-S2-S4-S2-S1、S1-S2-S4-S2-S2、S1-S2-S4-S2-S4、S1-S2-S4-S4-S1、S1-S2-S4-S4-S2、S1-S2-S4-S4-S4、S1-S4-S1-S1-S1、S1-S4-S1-S1-S2、S1-S4-S1-S1-S4、S1-S4-S1-S2-S1、S1-S4-S1-S2-S2、S1-S4-S1-S2-S4、S1-S4-S1-S4-S1、S1-S4-S1-S4-S2、S1-S4-S1-S4-S4、S1-S4-S2-S1-S1、S1-S4-S2-S1-S2、S1-S4-S2-S1-S4、S1-S4-S2-S2-S1、S1-S4-S2-S2-S2、S1-S4-S2-S2-S4、S1-S4-S2-S4-S1、S1-S4-S2-S4-S2、S1-S4-S2-S4-S4、S1-S4-S4-S1-S1、S1-S4-S4-S1-S2、S1-S4-S4-S1-S4、S1-S4-S4-S2-S1、S1-S4-S4-S2-S2、S1-S4-S4-S2-S4、S1-S4-S4-S4-S1、S1-S4-S4-S4-S2、S1-S4-S4-S4-S4、S2-S1-S1-S1-S1、S2-S1-S1-S1-S2、S2-S1-S1-S1-S4、S2-S1-S1-S2-S1、S2-S1-S1-S2-S2、S2-S1-S1-S2-S4、S2-S1-S1-S4-S1、S2-S1-S1-S4-S2、S2-S1-S1-S4-S4、S2-S1-S2-S1-S1、S2-S1-S2-S1-S2、S2-S1-S2-S1-S4、S2-S1-S2-S2-S1、S2-S1-S2-S2-S2、S2-S1-S2-S2-S4、S2-S1-S2-S4-S1、S2-S1-S2-S4-S2、S2-S1-S2-S4-S4、S2-S1-S4-S1-S1、S2-S1-S4-S1-S2、S2-S1-S4-S1-S4、S2-S1-S4-S2-S1、S2-S1-S4-S2-S2、S2-S1-S4-S2-S4、S2-S1-S4-S4-S1、S2-S1-S4-S4-S2、S2-S1-S4-S4-S4、S2-S2-S1-S1-S1、S2-S2-S1-S1-S2、S2-S2-S1-S1-S4、S2-S2-S1-S2-S1、S2-S2-S1-S2-S2、S2-S2-S1-S2-S4、S2-S2-S1-S4-S1、S2-S2-S1-S4-S2、S2-S2-S1-S4-S4、S2-S2-S2-S1-S1、S2-S2-S2-S1-S2、S2-S2-S2-S1-S4、S2-S2-S2-S2-S1、S2-S2-S2-S2-S2、S2-S2-S2-S2-S4、S2-S2-S2-S4-S1、S2-S2-S2-S4-S2、S2-S2-S2-S4-S4、S2-S2-S4-S1-S1、S2-S2-S4-S1-S2、S2-S2-S4-S1-S4、S2-S2-S4-S2-S1、S2-S2-S4-S2-S2、S2-S2-S4-S2-S4、S2-S2-S4-S4-S1、S2-S2-S4-S4-S2、S2-S2-S4-S4-S4、S2-S4-S1-S1-S1、S2-S4-S1-S1-S2、S2-S4-S1-S1-S4、S2-S4-S1-S2-S1、S2-S4-S1-S2-S2、S2-S4-S1-S2-S4、S2-S4-S1-S4-S1、S2-S4-S1-S4-S2、S2-S4-S1-S4-S4、S2-S4-S2-S1-S1、S2-S4-S2-S1-S2、S2-S4-S2-S1-S4、S2-S4-S2-S2-S1、S2-S4-S2-S2-S2、S2-S4-S2-S2-S4、S2-S4-S2-S4-S1、S2-S4-S2-S4-S2、S2-S4-S2-S4-S4、S2-S4-S4-S1-S1、S2-S4-S4-S1-S2、S2-S4-S4-S1-S4、S2-S4-S4-S2-S1、S2-S4-S4-S2-S2、S2-S4-S4-S2-S4、S2-S4-S4-S4-S1、S2-S4-S4-S4-S2、S2-S4-S4-S4-S4、S4-S1-S1-S1-S1、S4-S1-S1-S1-S2、S4-S1-S1-S1-S4、S4-S1-S1-S2-S1、S4-S1-S1-S2-S2、S4-S1-S1-S2-S4、S4-S1-S1-S4-S1、S4-S1-S1-S4-S2、S4-S1-S1-S4-S4、S4-S1-S2-S1-S1、S4-S1-S2-S1-S2、S4-S1-S2-S1-S4、S4-S1-S2-S2-S1、S4-S1-S2-S2-S2、S4-S1-S2-S2-S4、S4-S1-S2-S4-S1、S4-S1-S2-S4-S2、S4-S1-S2-S4-S4、S4-S1-S4-S1-S1、S4-S1-S4-S1-S2、S4-S1-S4-S1-S4、S4-S1-S4-S2-S1、S4-S1-S4-S2-S2、S4-S1-S4-S2-S4、S4-S1-S4-S4-S1、S4-S1-S4-S4-S2、S4-S1-S4-S4-S4、S4-S2-S1-S1-S1、S4-S2-S1-S1-S2、S4-S2-S1-S1-S4、S4-S2-S1-S2-S1、S4-S2-S1-S2-S2、S4-S2-S1-S2-S4、S4-S2-S1-S4-S1、S4-S2-S1-S4-S2、S4-S2-S1-S4-S4、S4-S2-S2-S1-S1、S4-S2-S2-S1-S2、S4-S2-S2-S1-S4、S4-S2-S2-S2-S1、S4-S2-S2-S2-S2、S4-S2-S2-S2-S4、S4-S2-S2-S4-S1、S4-S2-S2-S4-S2、S4-S2-S2-S4-S4、S4-S2-S4-S1-S1、S4-S2-S4-S1-S2, S4-S2-S4-S1-S4, S4-S2-S4-S2-S1, S4-S2-S4-S2-S2, S4-S2-S4-S2-S 4, S4-S2-S4-S4-S1, S4-S2-S4-S4-S2, S4-S2-S4-S4-S4, S4-S4-S1-S1-S1, S4-S4-S1-S1 -S2, S4-S4-S1-S1-S4, S4-S4-S1-S2-S1, S4-S4-S1-S2-S2, S4-S4-S1-S2-S4, S4-S4-S1- S4-S1, S4-S4-S1-S4-S2, S4-S4-S1-S4-S4, S4-S4-S2-S1-S1, S4-S4-S2-S1-S2, S4-S4-S2 -S1-S4, S4-S4-S2-S2-S1, S4-S4-S2-S2-S2, S4-S4-S2-S2-S4, S4-S4-S2-S4-S1, S4-S4- S2-S4-S2, S4-S4-S2-S4-S4, S4-S4-S4-S1-S1, S4-S4-S4-S1-S2, S4-S4-S4-S1-S4, S4-S and S4-S4-S4-S4-S4-S2-S4, S4-S4-S4-S2-S4, S4-S4-S4-S4-S1, S4-S4-S4-S4-S2, S4-S4-S4-S4-S4, or S4-S4-S4-S4-S4-S4, where S1, S2, S3, and S4 are different types of sugar modifications. In some embodiments, S1, S2, S3, and S4 are different types of 2'-sugar modifications. In some embodiments, these sugar modification patterns are present in the first wing, second wing, and / or core of oligonucleotides having an asymmetric format.

[0181] In some embodiments, the pattern of sugar modifications of oligonucleotides having an asymmetric format is one of the following: LLLLL, LLLLD, LLLDL, LLLDD, LLDLL, LLDLD, LLDDL, LLDDD, LDDLL, LDDLD, LDDDL, LDDDD, DLLLL, DLLLD, DLLDL, DLLDD, DLDLL, DLDLD, DLDDL, DLDDD, DDLLL, DDLLD, DDLDL, DDLDD, DDDLL, DDDDL, DDDDD, LLLLL, LLLLM, LLLML, LLLMM, LLMLL, LLMLM, LLMML, LLMMM, LMLLL, LMLLM, LMLML, LMLMM, LMMLL, LMMLM, LMMML, LMMMM, MLLLL, MLLLM, MLLML, MLLMM, MLMLL, MLMLM, MLMML , MLMMM, MMLLL, MMLLM, MMLML, MMLMM, MMMLL, MMMLM, MMMML, MMMMM, LLLLm, LLLmL, LLLmm, LLmLL, LLmLm, LLmmL, LLmmm, LmLLL, LmLLm, LmLmL, LmLmm, LmL L, LmmLm, LmmmL, Lmmmm, mLLLL, mLLLm, mLLmL, mLLmm, mLmLL, mLmLm, mLmL, mLmmm, mmLLL, mmLLm, mmLmL, mmLmm, mmmLL, mmmLm, mmmL, mmmmm, LLLDM, LLL ML, LLLMD, LLLMM, LLDLL, LLDLD, LLDLM, LLDDL, LLDDD, LLDDM, LLDML, LLDMD, LLDMM, LLMLL, LLMLD, LLMLM, LLMDL, LLMDD, LLMDM, LLMML, LLMMD, LLMMM, LD LLL, LDLLD, LDLLM, LDLDL, LDLDD, LDLDM, LDLML, LDLMD, LDLMM, LDDLL, LDDLD, LDDLM, LDDDL, LDDDD, LDDDM, LDDML, LDDMD, LDDMM, LDMLL, LDMLD, LDMLM, L DMDL, LDMDD, LDMDM, LDMML, LDMMD, LDMMM, LMLLL, LMLLD, LMLLM, LMLDL, LMLDD, LMLDM, LMLML, LMLMD, LMLMM, LMDLL, LMDLD, LMDLM, LMDDL, LMDDD, LMDDM,LMDML、LMDMD、LMDMM、LMMLL、LMMLD、LMMLM、LMMDL、LMMDD、LMMDM、LMMML、LMMMD、LMMMM、DLLLL、DLLLD、DLLLM、DLLDL、DLLDD、DLLDM、DLLML、DLLMD、DLLMM、DLDLL、DLDLD、DLDLM、DLDDL、DLDDD、DLDDM、DLDML、DLDMD、DLDMM、DLMLL、DLMLD、DLMLM、DLMDL、DLMDD、DLMDM、DLMML、DLMMD、DLMMM、DDLLL、DDLLD、DDLLM、DDLDL、DDLDD、DDLDM、DDLML、DDLMD、DDLMM、DDDLL、DDDLD、DDDLM、DDDDL、DDDDD、DDDDM、DDDML、DDDMD、DDDMM、DDMLL、DDMLD、DDMLM、DDMDL、DDMDD、DDMDM、DDMML、DDMMD、DDMMM、DMLLL、DMLLD、DMLLM、DMLDL、DMLDD、DMLDM、DMLML、DMLMD、DMLMM、DMDLL、DMDLD、DMDLM、DMDDL、DMDDD、DMDDM、DMDML、DMDMD、DMDMM、DMMLL、DMMLD、DMMLM、DMMDL、DMMDD、DMMDM、DMMML、DMMMD、DMMMM、MLLLL、MLLLD、MLLLM、MLLDL、MLLDD、MLLDM、MLLML、MLLMD、MLLMM、MLDLL、MLDLD、MLDLM、MLDDL、MLDDD、MLDDM、MLDML、MLDMD、MLDMM、MLMLL、MLMLD、MLMLM、MLMDL、MLMDD、MLMDM、MLMML、MLMMD、MLMMM、MDLLL、MDLLD、MDLLM、MDLDL、MDLDD、MDLDM、MDLML、MDLMD、MDLMM、MDDLL、MDDLD、MDDLM、MDDDL、MDDDD、MDDDM、MDDML、MDDMD、MDDMM、MDMLL、MDMLD、MDMLM、MDMDL、MDMDD、MDMDM、MDMML、MDMMD、MDMMM、MMLLL、MMLLD、MMLLM、MMLDL、MMLDD、MMLDM、MMLML、MMLMD、MMLMM、MMDLL、MMDLD、MMDLM、MMDDL、MMDDD、MMDDM、MMDML、MMDMD、MMDMM、MMMLL、MMMLD、MMMLM、MMMDL、MMMDD、MMMDM、MMMMML、MMMMD、MMMMMM、MMMMMM、LLLDm、LL LmL、LLLmD、LLLmm、LLDLL、LLDLD、LLDLm、LLDDL、LLDDD、LLDDm、LLDmL、LLDm D, LLDmm, LLLmLL, LLLmLD, LLLmLm, LLLmDL, LLLmDD, LLLmDm, LLLmmL, LLLmmD, LLLmmm, LLDLLL, LLDLLD, LLDLLm, LLDLDL, LLDLDD, LLDLDm, LLDLmL, LLDLmD, LLDLmm, LLDDLL, LLDD LD、LDDLm、LDDDL、LDDDD、LDDDm、LDDmL、LDDmD、LDDmm、LDmLL、LDmLD、LDmLm 、LDmDL、LDmDD、LDmDm、LDmmL、LDmmD、LDmmm、LmLLL、LmLLD、LmLLm、LMLDL、L mLDD、LmLDm、LmLmL、LmLmD、LmLmm、LmDLL、LmDLD、LmDLm、LmDDL、LmDDD、LmD Dm、LmDmL、LmDmD、LmDmm、LmmLL、LmmLD、LmmLm、LmmDL、LmmDD、LmmDm、LmmmL、 LmmmD、Lmmmm、DLLLL、DLLLD、DLLLm、DLLDL、DLLDD、DLLDm、DLLmL、DLLmD、DL Lmm、DLDLL、DLDLD、DLDLm、DLDDL、DLDDD、DLDDm、DLDmL、DLDmD、DLDmm、DLmL L、DLmLD、DLmLm、DLmDL、DLmDD、DLmDm、DLmmL、DLmmD、DLmmm、DDLLL、DDLLD、 DDLLm、DDLDL、DDLDD、DDLDm、DDLmL、DDLmD、DDLmm、DDDLL、DDDLD、DDDLm、DDD DL、DDDDD、DDDDm、DDDmL、DDDmD、DDDmm、DDmLL、DDmLD、DDmLm、DDmDL、DDmDD 、DDmDm、DDmmL、DDmmD、DDmmm、DmLLL、DmLLD、DmLLm、DmLDL、DmLDD、DmLDm、D mLmL、DmLmD、DmLmm、DmDLL、DmDLD、DmDLm、DmDDL、DmDDD、DmDDm、DmDmL、DmD mD、DmDmm、DmmLL、DmmLD、DmmLm、DmmDL、DmmDD、DmmDm、DmmmL、DmmmD、Dmmmm、mLLLL, mLLLD, mLLLm, mLLDL, mLLDD, mLLDm, mLLmL, mLLmD, mLLmm, mLDLL, mLDLD, mLDLm, mLDDL, mLDDD, mLDDm, mLDmL, mLDmD, mLDmm, mLmLL, mLmLD, mLmLm, mLmDL, mLmD D, mLmDm, mLmmL, mLmmD, mLmmm, mDLLL, mDLLD, mDLLm, mDLDL, mDLDD, mDLDm, mDLmL, mDLmD, mDLmm, mDDLL, mDDLD, mDDLm, mDDDL, mDDDD, mDDDm, mDDmL, mDDmD, mDDmm, mD mLL, mDmLD, mDmLm, mDmDL, mDmDD, mDmDm, mDmmL, mDmmD, mDmmm, mmLLL, mmLLD, mmLLm, mmLDL, mmLDD, mmLDm, mmLmL, mmLmD, mmLmm, mmDLL, mmDLD, mmDLm, mmDDL, mmDDD, mmDDm, mmDmL, mmDmD, mmDmm, mmmLL, mmmLD, mmmLm, mmmDL, mmmDD, mmmDm, mmmmL, mmmmD, mmmmm, or mmmmm, where L=LNA, D=deoxy, M=2'-MOE, and m=2'-OMe. In some embodiments, such sugar modification patterns are present in the first wing, second wing, and / or core of oligonucleotides having an asymmetric format.

[0182] Various non-limiting examples of asymmetric formats of oligonucleotides are described herein, as well as various non-limiting examples of oligonucleotides having such formats.

[0183] In some embodiments of oligonucleotides having an asymmetric format, the core comprises any sugar or sugar modification described herein or known in the art, or any pattern or combination of two or more different sugars and / or sugar modifications.

[0184] In some embodiments of oligonucleotides having an asymmetric format, the core comprises a D. Non-limiting examples of such oligonucleotides include the following: WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, WV-8664.

[0185] In some embodiments of oligonucleotides having an asymmetric format, the core comprises DD. Non-limiting examples of such oligonucleotides include the following: WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, WV-8664.

[0186] In some embodiments of oligonucleotides having an asymmetric format, the core comprises DDD. Non-limiting examples of such oligonucleotides include the following: WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, WV-8664.

[0187] In some embodiments of oligonucleotides having an asymmetric format, the core comprises DDDD. Non-limiting examples of such oligonucleotides include the following: WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, WV-8664.

[0188] In some embodiments of oligonucleotides having an asymmetric format, the core comprises DDDDD. Non-limiting examples of such oligonucleotides include the following: WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, WV-8664.

[0189] In some embodiments of oligonucleotides having an asymmetric format, the core comprises DDDDDD. Non-limiting examples of such oligonucleotides include the following: WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, WV-8664.

[0190] In some embodiments of oligonucleotides having an asymmetric format, the core comprises DDDDDDD. Non-limiting examples of such oligonucleotides include the following: WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, WV-8664.

[0191] In some embodiments of oligonucleotides having an asymmetric format, the core comprises DDDDDDDD. Non-limiting examples of such oligonucleotides include the following: WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, WV-8664.

[0192] In some embodiments of oligonucleotides having an asymmetric format, the core comprises DDDDDDDDDD. Non-limiting examples of such oligonucleotides include the following: WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, WV-8664.

[0193] In some embodiments of oligonucleotides having an asymmetric format, the core comprises DDDDDDDDDD. Non-limiting examples of such oligonucleotides include the following: WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, WV-8664.

[0194] In some embodiments of oligonucleotides having an asymmetric format, the core comprises a 5' BrdU.

[0195] In some embodiments of oligonucleotides having an asymmetric format, the core comprises a 5' BrdU and a D.

[0196] In some embodiments of oligonucleotides having an asymmetric format, the core comprises a 5' BrdU and two or more Ds.

[0197] In some embodiments of oligonucleotides having an asymmetric format, the core comprises a 5' BrdU and two or more consecutive Ds.

[0198] In some embodiments, oligonucleotides having an asymmetric format can comprise a first wing; and a second wing, wherein the first and second wings are chemically distinct from each other and from the core.

[0199] Non-limiting examples of oligonucleotide formats having asymmetric formats in which the first and second wings are chemically distinct from each other and from the core are shown in Figures 1A and 1B, and the legends for Figures 1A and 1B are provided in Figure 1D.

[0200] In some embodiments, oligonucleotides having an asymmetric format can comprise a first wing; and a second wing, wherein the first and second wings are chemically distinct from each other and from the core, and the first and / or second wing can comprise M, m, and / or L, where M is (at least one) 2'-MOE (or optionally methyl-C2'-MOE if the base is C), m is (at least one) 2'-OMe, and L is (at least one) LNA, wherein either the first or second wing can be located at the 5'-end of the wing-core-wing format, with the other wing located at the 3'-end.

[0201] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; the second wing comprises two or more consecutive m's. Non-limiting examples of such oligonucleotides include WV-8852 and WV-8856.

[0202] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises two or more consecutive M's. Non-limiting examples of such oligonucleotides include WV-8043-8048.

[0203] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mmmmm: the second wing comprises MMMMM. Non-limiting examples of such oligonucleotides include WV-8043-8048.

[0204] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises MMMMM: the second wing comprises mmmmm. Non-limiting examples of such oligonucleotides include WV-8852 and WV-8856.

[0205] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a particular order and number of m and M; and the second wing comprises a different order and / or number of m and M. A non-limiting example of such an oligonucleotide is WV-8248.

[0206] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMmMm; and the second wing comprises mMmmm. A non-limiting example of such an oligonucleotide is WV-8248.

[0207] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises an m and two or more consecutive M's; and the second wing comprises an M and two or more consecutive m's. Non-limiting examples of such oligonucleotides include WV-9894-9896.

[0208] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMM; and the second wing comprises Mmmmm. Non-limiting examples of such oligonucleotides include WV-9894-9896 and WV-10253-10254.

[0209] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises m and M in a particular order and number; the second wing comprises two or more consecutive Ms.

[0210] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMMMM; and the second wing comprises MMMMMMM. Non-limiting examples of such oligonucleotides include WV-12099, WV-12101, WV-12103, WV-12105, WV-12107, and WV-12109.

[0211] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMM; and the second wing comprises MMMMM. Non-limiting examples of such oligonucleotides include WV-10250 and WV-9869-9870.

[0212] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises all M. Non-limiting examples of such oligonucleotides include WV-9441-9445.

[0213] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises MMMMM. Non-limiting examples of such oligonucleotides include WV-9441-9445.

[0214] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises m and M in a particular order and number; the second wing comprises two or more consecutive m's.

[0215] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; the second wing comprises m's and M's in a particular order and number.

[0216] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; the second wing comprises m's and M's in a particular order and number.

[0217] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises L, m, and M in a particular order and number; and the second wing comprises L, m, and M in a different order and / or number. A non-limiting example of such an oligonucleotide is WV-8250.

[0218] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises LMmMm; and the second wing comprises mMmmL. A non-limiting example of such an oligonucleotide is WV-8250.

[0219] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises L, m, and M; and the second wing comprises m and L. A non-limiting example of such an oligonucleotide is WV-8246.

[0220] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises LMmMm; and the second wing comprises mmmmL. A non-limiting example of such an oligonucleotide is WV-8246.

[0221] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises L and M; and the second wing comprises m and L. Non-limiting examples of such oligonucleotides include WV-11958, and WV-11960, and WV-11962.

[0222] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises an L and two or more consecutive M's; and the second wing comprises two or more consecutive m's and an L. Non-limiting examples of such oligonucleotides include WV-11958, WV-11960, and WV-11962.

[0223] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises LMMMM; and the second wing comprises mmmmL. Non-limiting examples of such oligonucleotides include WV-11958, WV-11960, and WV-11962.

[0224] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications; and the second wing comprises only one type of sugar modification. Non-limiting examples of such oligonucleotides include WV-11114, WV-11533, WV-12503, WV-12504, WV-12505, WV-8553, WV-8555, WV-8556, WV-8557, WV-8582, WV-8583, WV-8584, WV-8585, WV-8586, WV-8587, WV-8588, WV-8589, WV-8590, WV-8591, WV-8592, WV-8593, WV-9058, WV-9059, WV-9060, WV-9061, WV-9696, WV-9697, and WV-9698.

[0225] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a pattern of m and M; and the second wing comprises two or more consecutive m's. Non-limiting examples of such oligonucleotides include WV-11114, WV-11533, WV-12503, WV-12504, WV-12505, WV-8553, WV-8555, WV-8556, WV-8557, WV-8582, WV-8583, WV-8584, WV-8585, WV-8586, WV-8587, WV-8588, WV-8589, WV-8590, WV-8591, WV-8592, WV-8593, WV-9058, WV-9059, WV-9060, WV-9061, WV-9696, WV-9697, and WV-9698.

[0226] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises two or more consecutive m's. Non-limiting examples of such oligonucleotides include WV-11114, WV-11533, WV-12503, WV-12504, WV-12505, WV-8553, WV-8555, WV-8556, WV-8557, WV-8582, WV-8583, WV-8584, WV-8585, WV-8586, WV-8587, WV-8588, WV-8589, WV-8590, WV-8591, WV-8592, WV-8593, WV-9058, WV-9059, WV-9060, WV-9061, WV-9696, WV-9697, and WV-9698.

[0227] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a pattern of two different types of sugar modifications; and the second wing comprises a pattern of the same two different types of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8097, WV-8098, WV-8099, WV-8100, WV-8101, WV-8102, and WV-8109.

[0228] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a pattern of m and M; and the second wing comprises a different pattern of m and M. Non-limiting examples of such oligonucleotides include WV-8097, WV-8098, WV-8099, WV-8100, WV-8101, WV-8102, and WV-8109.

[0229] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mmMMm; and the second wing comprises mmMmm. Non-limiting examples of such oligonucleotides include WV-8097, WV-8098, WV-8099, WV-8100, WV-8101, WV-8102, and WV-8109.

[0230] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises an m and two or more consecutive M's; and the second wing comprises two or more consecutive m's. Non-limiting examples of such oligonucleotides include WV-12110, WV-12111, WV-12112, WV-12113, and WV-12114.

[0231] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMM; and the second wing comprises two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-12110, WV-12111, WV-12112, WV-12113, and WV-12114.

[0232] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises two or more consecutive m's. Non-limiting examples of such oligonucleotides include WV-13303, WV-13304, WV-13809, WV-14087, WV-14349, WV-14556, WV-14557, WV-14558, WV-14559, WV-14560, WV-14561, WV-14562, WV-14563, WV-14564, WV-14733, WV-14734, WV-14735, WV-14736, WV-14737, WV-14771, WV-15310, WV-15311, WV-15312, WV-15313, WV-15314, WV-15315, WV-15316, WV-15317, WV-15318, WV-15319, WV-15320, WV-15321 , WV-15351, WV-15352, WV-15353, WV-15354, WV-15355, WV-15356, WV-15357 , WV-15358, WV-15359, WV-15360, WV-15361, WV-15362, WV-15363, WV-1536 4, WV-15365, WV-15562, WV-15563, WV-15863, WV-15864 and WV-15887.

[0233] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises two or more consecutive m's. Non-limiting examples of such oligonucleotides include WV-8552, WV-8554, WV-8570, WV-8571, WV-8572, WV-8573, WV-8574, WV-8575, WV-8576, WV-8577, WV-8578, WV-8579, WV-8580, and WV-8581.

[0234] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises two different types of sugar modifications. Non-limiting examples of such oligonucleotides include WV-14552, WV-14553, WV-14554, and WV-14555.

[0235] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises two different types of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8456.

[0236] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises two or more consecutive m's. Non-limiting examples of such oligonucleotides include WV-8005, WV-8006, WV-8007, WV-8008, WV-8466, WV-8467, WV-8468, WV-8469, WV-8470, WV-8471, WV-8547, WV-8548, WV-8594, WV-13305, WV-13306, WV-13307, WV-13308, WV-13309, WV-13310, WV-13311, WV-13313, WV-13803, WV-13804, and WV-13805.

[0237] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises two different types of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8125.

[0238] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises two or more consecutive M's. Non-limiting examples of such oligonucleotides include WV-8314.

[0239] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modification and the second wing comprises a third type of sugar modification. Non-limiting examples of such oligonucleotides include WV-9508, WV-9509, and WV-9510.

[0240] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications; and the second wing comprises two or more consecutive F. Non-limiting examples of such oligonucleotides include WV-9508, WV-9509, and WV-9510.

[0241] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications; and the second wing comprises two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-8595, WV-8691, WV-8692, WV-8693, WV-8694, WV-8695, WV-8696, WV-9062, WV-9063, WV-9285, WV-9286, WV-9380, WV-9381, WV-9394, WV-9395, WV-9396, WV-9397, WV-9398, WV-9399, WV-9421, WV-9421, WV-9486, and WV-9487.

[0242] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications; and the second wing comprises two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-9488, WV-9489, WV-9490, WV-9491, WV-9492, WV-9494, WV-9505, WV-9506, WV-9507, WV-8452, WV-8453, WV-8009, WV-8010, WV-8011, WV-8012, WV-8454, and WV-8455.

[0243] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications; and the second wing comprises two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-8549, WV-8550, WV-8551, WV-8568, WV-8569, WV-13312, WV-14758, WV-14772, WV-15049, WV-15050, and WV-15051.

[0244] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a particular pattern of two different types of sugar modifications; and the second wing comprises another pattern of two different types of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8121, WV-8129, WV-8118, WV-8126, WV-8472, WV-8473, WV-8474, WV-8475, and WV-8476.

[0245] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a particular pattern of two different types of sugar modifications; the second wing comprises another pattern of two different types of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8119 and WV-8127.

[0246] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modification; the second wing comprises only one type of sugar modification. Non-limiting examples of such oligonucleotides include WV-8115 and WV-8123.

[0247] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a particular pattern of two different types of sugar modifications; the second wing comprises another pattern of two different types of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8120 and WV-8128.

[0248] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications; the second wing comprises only one type of sugar modification. Non-limiting examples of such oligonucleotides include WV-8116 and WV-8124.

[0249] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises an M and two or more consecutive m's. Non-limiting examples of such oligonucleotides include WV-14552, WV-14553, WV-14554, and WV-14555.

[0250] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises MmMmm. A non-limiting example of such an oligonucleotide is WV-8456.

[0251] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises two or more consecutive m's. Non-limiting examples of such oligonucleotides include WV-8005, WV-8006, WV-8007, WV-8008, WV-8466, WV-8467, WV-8468, WV-8469, WV-8470, WV-8471, WV-8547, WV-8548, WV-8594, WV-13305, WV-13306, WV-13307, WV-13308, WV-13309, WV-13310, WV-13311, WV-13313, WV-13803, WV-13804, and WV-13805.

[0252] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises an M and two or more consecutive m's. Non-limiting examples of such oligonucleotides include WV-8125.

[0253] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises two or more consecutive M's. Non-limiting examples of such oligonucleotides include WV-8314.

[0254] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modification and the second wing comprises a third type of sugar modification. Non-limiting examples of such oligonucleotides include WV-9508, WV-9509, and WV-9510.

[0255] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises two or more consecutive F. Non-limiting examples of such oligonucleotides include WV-9508, WV-9509, and WV-9510.

[0256] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-8595, WV-8691, WV-8692, WV-8693, WV-8694, WV-8695, WV-8696, WV-9062, WV-9063, WV-9285, WV-9286, WV-9380, WV-9381, WV-9394, WV-9395, WV-9396, WV-9397, WV-9398, WV-9399, WV-9421, WV-9421, WV-9486, and WV-9487.

[0257] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-9488, WV-9489, WV-9490, WV-9491, WV-9492, WV-9494, and WV-9505, WV-9506, and WV-9507.

[0258] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a particular pattern of two different types of sugar modifications; the second wing comprises another pattern of two different types of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8452 and WV-8453.

[0259] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises MmMmm. Non-limiting examples of such oligonucleotides include WV-8452 and WV-8453.

[0260] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-8009, WV-8010, WV-8011, WV-8012, WV-8454, and WV-8455.

[0261] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-8549, WV-8550, WV-8551, WV-8568, WV-8569, WV-13312, WV-14758, WV-14772, WV-15049, WV-15050, and WV-15051.

[0262] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMmm; and the second wing comprises an M and two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-8121 and WV-8129.

[0263] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications and the second wing comprises only one type of sugar modification. Non-limiting examples of such oligonucleotides include WV-8118, WV-8126, WV-8472, WV-8473, WV-8474, WV-8475, and WV-8476.

[0264] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMmm; and the second wing comprises two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-8118, WV-8126, WV-8472, WV-8473, WV-8474, WV-8475, and WV-8476.

[0265] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMmmm; and the second wing comprises an M and two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-8119 and WV-8127.

[0266] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMmmm; and the second wing comprises two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-8115 and WV-8123.

[0267] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mmMmm; and the second wing comprises an M and two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-8120 and WV-8128.

[0268] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mmMmm; and the second wing comprises two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-8116 and WV-8124.

[0269] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises DMMD. Non-limiting examples of such oligonucleotides include WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, and WV-8664.

[0270] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises an MMD. Non-limiting examples of such oligonucleotides include WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, and WV-8664.

[0271] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises DDMMD. Non-limiting examples of such oligonucleotides include WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, and WV-8664.

[0272] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises two or more consecutive m's. Non-limiting examples of such oligonucleotides include WV-8637, WV-8638, WV-8639, WV-8640, WV-8653, WV-8654, WV-8655, WV-8656, WV-8665, WV-8666, WV-8667, WV-8668, WV-8669, WV-8670, WV-8671, WV-8672, WV-12947, WV-12948, WV-12949, WV-12950, ​​WV-12951, WV-12952, WV-12953 , WV-12954, WV-12955, WV-12956, WV-12957, WV-12958, WV-12959, WV-12960, WV-12961, WV-12962, WV-12963, WV-12964, WV-1296 5, WV-12966, WV-12967, WV-12968, WV-12969, WV-12970, WV-12971, WV-12972, WV-12973, WV-12974, WV-12975 and WV-12976.

[0273] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises two or more consecutive M's. Non-limiting examples of such oligonucleotides include WV-12977, WV-12978, WV-12979, WV-12980, WV-12981, WV-12982, WV-12983, WV-12984, WV-12985, WV-12986, WV-12987, WV-12988, WV-12989, WV-12990, WV-12991, Examples include WV-12992, WV-12993, WV-12994, WV-12995, WV-12996, WV-12997, WV-12998, WV-12999, WV-13000, WV-13001, WV-13002, WV-13003, WV-13004, WV-13005, WV-13006, WV-13007 and WV-13008.

[0274] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMM; and the second wing comprises an M and two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-9887, WV-9888, WV-10245, and WV-10246.

[0275] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMM; and the second wing comprises two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-9871 and WV-9872.

[0276] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMMMM; and the second wing comprises mmmmmmm. A non-limiting example of such an oligonucleotide is WV-12101.

[0277] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMM; and the second wing comprises an M and two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-9873 and WV-9874.

[0278] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMM; and the second wing comprises two or more consecutive m. Non-limiting examples of such oligonucleotides include WV-9885, WV-9886, WV-10243, and WV-10244.

[0279] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises two or more consecutive F. Non-limiting examples of such oligonucleotides include WV-9526, WV-9527, WV-9528, WV-9529, WV-9530, WV-9531, WV-9532, WV-9533, WV-9590, WV-9591, WV-9592, and WV-9593.

[0280] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises two or more consecutive m's. Non-limiting examples of such oligonucleotides include WV-8610, WV-8611, WV-8612, WV-8613, WV-8614, WV-8615, WV-8616, WV-8617, WV-8618, WV-8619, WV-8629, WV-8632, WV-8673, WV-8674, WV-8675, WV-8676, WV-8677, WV-8678, WV-8679, WV-8680, WV-8681, WV-8682, WV-8683, WV-8684, WV-8685, WV-8686, WV-8687, and WV-8688.

[0281] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8852 and WV-8856.

[0282] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8043-8048.

[0283] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises MMMMM; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8043-8048.

[0284] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mmmmm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8852 and WV-8856.

[0285] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a different order and / or number of m and M; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-8248.

[0286] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMmmm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-8248.

[0287] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises an M and two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9894 to 9896.

[0288] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises Mmmmm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9894-9896 and WV-10253-10254.

[0289] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications.

[0290] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises MMMMMMM; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-12099, WV-12101, WV-12103, WV-12105, WV-12107, and WV-12109.

[0291] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises MMMMM; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-10250 and WV-9869-WV-9870.

[0292] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises all M; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9441 to -9445.

[0293] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises MMMMM; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9441 to -9445.

[0294] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications.

[0295] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a particular order and number of m and M; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications.

[0296] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a particular order and number of m and M; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications.

[0297] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a different order and / or number of L, m, and M; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-8250.

[0298] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMmmL; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-8250.

[0299] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises m and L; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-8246.

[0300] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mmmmL; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-8246.

[0301] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises m and L; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-11958, WV-11960, and WV-11962.

[0302] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m and L; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-11958, WV-11960, and WV-11962.

[0303] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mmmmL; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-11958, WV-11960, and WV-11962.

[0304] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises only one type of sugar modification; the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-11114, WV-11533, WV-12503, WV-12504, WV-12505, WV-8553, WV-8555, WV-8556, WV-8557, WV-8582, WV-8583, WV-8584, WV-8585, WV-8586, WV-8587, WV-8588, WV-8589, WV-8590, WV-8591, WV-8592, WV-8593, WV-9058, WV-9059, WV-9060, WV-9061, WV-9696, WV-9697, and WV-9698.

[0305] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-11114, WV-11533, WV-12503, WV-12504, WV-12505, WV-8553, WV-8555, WV-8556, WV-8557, WV-8582, WV-8583, WV-8584, WV-8585, WV-8586, WV-8587, WV-8588, WV-8589, WV-8590, WV-8591, WV-8592, WV-8593, WV-9058, WV-9059, WV-9060, WV-9061, WV-9696, WV-9697, and WV-9698.

[0306] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-11114, WV-11533, WV-12503, WV-12504, WV-12505, WV-8553, WV-8555, WV-8556, WV-8557, WV-8582, WV-8583, WV-8584, WV-8585, WV-8586, WV-8587, WV-8588, WV-8589, WV-8590, WV-8591, WV-8592, WV-8593, WV-9058, WV-9059, WV-9060, WV-9061, WV-9696, WV-9697, and WV-9698.

[0307] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a different pattern of the same two different types of sugar modifications; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8097, WV-8098, WV-8099, WV-8100, WV-8101, WV-8102, and WV-8109.

[0308] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a different pattern of m and M; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8097, WV-8098, WV-8099, WV-8100, WV-8101, WV-8102, and WV-8109.

[0309] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mmMmm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8097, WV-8098, WV-8099, WV-8100, WV-8101, WV-8102, and WV-8109.

[0310] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-12110, WV-12111, WV-12112, WV-12113, and WV-12114.

[0311] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-12110, WV-12111, WV-12112, WV-12113, and WV-12114.

[0312] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-13303, WV-13304, WV-13809, WV-14087, WV-14349, WV-14556, WV-14557, WV-14558, WV-14559, WV-14560, WV-14561, WV-14562, WV-14563, WV-14564, WV-14733, WV-14734, WV-14735, WV-14736, WV-14737, WV-14771, WV-15310, WV-15311, WV-15312, WV-15313, WV-15314, WV-15315, WV-15316, WV-15317, WV-15318, WV-15319, WV-15320, WV-15321 , WV-15351, WV-15352, WV-15353, WV-15354, WV-15355, WV-15356, WV-15357 , WV-15358, WV-15359, WV-15360, WV-15361, WV-15362, WV-15363, WV-1536 4, WV-15365, WV-15562, WV-15563, WV-15863, WV-15864 and WV-15887.

[0313] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8552, WV-8554, WV-8570, WV-8571, WV-8572, WV-8573, WV-8574, WV-8575, WV-8576, WV-8577, WV-8578, WV-8579, WV-8580, and WV-8581.

[0314] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-14552, WV-14553, WV-14554, and WV-14555.

[0315] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-8456.

[0316] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8005, WV-8006, WV-8007, WV-8008, WV-8466, WV-8467, WV-8468, WV-8469, WV-8470, WV-8471, WV-8547, WV-8548, WV-8594, WV-13305, WV-13306, WV-13307, WV-13308, WV-13309, WV-13310, WV-13311, WV-13313, WV-13803, WV-13804, and WV-13805.

[0317] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-8125.

[0318] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8314.

[0319] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a third type of sugar modification and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9508, WV-9509, and WV-9510.

[0320] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive F's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9508, WV-9509, and WV-9510.

[0321] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8595, WV-8691, WV-8692, WV-8693, WV-8694, WV-8695, WV-8696, WV-9062, WV-9063, WV-9285, WV-9286, WV-9380, WV-9381, WV-9394, WV-9395, WV-9396, WV-9397, WV-9398, WV-9399, WV-9421, WV-9421, WV-9486, and WV-9487.

[0322] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9488, WV-9489, WV-9490, WV-9491, WV-9492, WV-9494, WV-9505, WV-9506, WV-9507, WV-8452, WV-8453, WV-8009, WV-8010, WV-8011, WV-8012, WV-8454, and WV-8455.

[0323] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8549, WV-8550, WV-8551, WV-8568, WV-8569, WV-13312, WV-14758, WV-14772, WV-15049, WV-15050, and WV-15051.

[0324] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a different pattern of two different types of sugar modifications; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8121, WV-8129, WV-8118, WV-8126, WV-8472, WV-8473, WV-8474, WV-8475, and WV-8476.

[0325] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a different pattern of two different types of sugar modifications; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8119 and WV-8127.

[0326] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises only one type of sugar modification and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8115 and WV-8123.

[0327] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a different pattern of two different types of sugar modifications; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8120 and WV-8128.

[0328] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises one type of sugar modification and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8116 and WV-8124.

[0329] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises an M and two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-14552, WV-14553, WV-14554, and WV-14555.

[0330] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises MmMmm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-8456.

[0331] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8005, WV-8006, WV-8007, WV-8008, WV-8466, WV-8467, WV-8468, WV-8469, WV-8470, WV-8471, WV-8547, WV-8548, WV-8594, WV-13305, WV-13306, WV-13307, WV-13308, WV-13309, WV-13310, WV-13311, WV-13313, WV-13803, WV-13804, and WV-13805.

[0332] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises an M and two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8125.

[0333] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8314.

[0334] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a third type of sugar modification and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9508, WV-9509, and WV-9510.

[0335] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive F's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9508, WV-9509, and WV-9510.

[0336] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8595, WV-8691, WV-8692, WV-8693, WV-8694, WV-8695, WV-8696, WV-9062, WV-9063, WV-9285, WV-9286, WV-9380, WV-9381, WV-9394, WV-9395, WV-9396, WV-9397, WV-9398, WV-9399, WV-9421, WV-9421, WV-9486, and WV-9487.

[0337] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9488, WV-9489, WV-9490, WV-9491, WV-9492, WV-9494, WV-9505, WV-9506, and WV-9507.

[0338] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a different pattern of two different types of sugar modifications; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8452 and WV-8453.

[0339] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises MmMmm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8452 and WV-8453.

[0340] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8009, WV-8010, WV-8011, WV-8012, WV-8454, and WV-8455.

[0341] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8549, WV-8550, WV-8551, WV-8568, WV-8569, WV-13312, WV-14758, WV-14772, WV-15049, WV-15050, and WV-15051.

[0342] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises an M and two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8121 and WV-8129.

[0343] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises only one type of sugar modification and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8118, WV-8126, WV-8472, WV-8473, WV-8474, WV-8475, and WV-8476.

[0344] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8118, WV-8126, WV-8472, WV-8473, WV-8474, WV-8475, and WV-8476.

[0345] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises an M and two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8119 and WV-8127.

[0346] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8115 and WV-8123.

[0347] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises an M and two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8120 and WV-8128.

[0348] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8116 and WV-8124.

[0349] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises DMMD; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, and WV-8664.

[0350] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises an MMD and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, and WV-8664.

[0351] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises DDMMD; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, and WV-8664.

[0352] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8637, WV-8638, WV-8639, WV-8640, WV-8653, WV-8654, WV-8655, WV-8656, WV-8665, WV-8666, WV-8667, WV-8668, WV-8669, WV-8670, WV-8671, WV-8672, WV-12947, WV-12948, WV-12949, WV-12950, ​​WV-12951, WV-12952, and WV-12953. , WV-12954, WV-12955, WV-12956, WV-12957, WV-12958, WV-12959, WV-12960, WV-12961, WV-12962, WV-12963, WV-12964, WV-1296 5, WV-12966, WV-12967, WV-12968, WV-12969, WV-12970, WV-12971, WV-12972, WV-12973, WV-12974, WV-12975 and WV-12976.

[0353] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-12977, WV-12978, WV-12979, WV-12980, WV-12981, WV-12982, WV-12983, WV-12984, WV-12985, WV-12986, WV-12987, WV-12988, WV-12989, WV-12990, WV-12991, Examples include WV-12992, WV-12993, WV-12994, WV-12995, WV-12996, WV-12997, WV-12998, WV-12999, WV-13000, WV-13001, WV-13002, WV-13003, WV-13004, WV-13005, WV-13006, WV-13007 and WV-13008.

[0354] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises an M and two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9887, WV-9888, WV-10245, and WV-10246.

[0355] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9871 and WV-9872.

[0356] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mmmmmmm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-12101.

[0357] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises an M and two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9873 and WV-9874.

[0358] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9885, WV-9886, WV-10243, and WV-10244.

[0359] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive F's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9526, WV-9527, WV-9528, WV-9529, WV-9530, WV-9531, WV-9532, WV-9533, WV-9590, WV-9591, WV-9592, and WV-9593.

[0360] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8610, WV-8611, WV-8612, WV-8613, WV-8614, WV-8615, WV-8616, WV-8617, WV-8618, WV-8619, WV-8629, WV-8632, WV-8673, WV-8674, WV-8675, WV-8676, WV-8677, WV-8678, WV-8679, WV-8680, WV-8681, WV-8682, WV-8683, WV-8684, WV-8685, WV-8686, WV-8687, and WV-8688.

[0361] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8852 and WV-8856.

[0362] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8043-8048.

[0363] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mmmmm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8043-8048.

[0364] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises MMMMM; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8852 and WV-8856.

[0365] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a particular order and number of m and M; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-8248.

[0366] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMmMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-8248.

[0367] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises m and two or more consecutive M's; the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9894 to 9896.

[0368] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMM; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9894-9896 and WV-10253-10254.

[0369] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a particular order and number of m and M; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications.

[0370] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMMMM; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-12099, WV-12101, WV-12103, WV-12105, WV-12107, and WV-12109.

[0371] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMM; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-10250 and WV-9869-WV-9870.

[0372] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9441-9445.

[0373] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9441-9445.

[0374] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a particular order and number of m and M; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications.

[0375] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications.

[0376] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications.

[0377] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a particular order and number of L, m, and M; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-8250.

[0378] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises LMmMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-8250.

[0379] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises L, m, and M; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-8246.

[0380] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises LMmMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-8246.

[0381] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises an L and an M; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-11958, WV-11960, and WV-11962.

[0382] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises an L and two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-11958, WV-11960, and WV-11962.

[0383] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises LMMMM; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-11958, WV-11960, and WV-11962.

[0384] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-11114, WV-11533, WV-12503, WV-12504, WV-12505, WV-8553, WV-8555, WV-8556, WV-8557, WV-8582, WV-8583, WV-8584, WV-8585, WV-8586, WV-8587, WV-8588, WV-8589, WV-8590, WV-8591, WV-8592, WV-8593, WV-9058, WV-9059, WV-9060, WV-9061, WV-9696, WV-9697, and WV-9698.

[0385] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises one pattern of m and M; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-11114, WV-11533, WV-12503, WV-12504, WV-12505, WV-8553, WV-8555, WV-8556, WV-8557, WV-8582, WV-8583, WV-8584, WV-8585, WV-8586, WV-8587, WV-8588, WV-8589, WV-8590, WV-8591, WV-8592, WV-8593, WV-9058, WV-9059, WV-9060, WV-9061, WV-9696, WV-9697, and WV-9698.

[0386] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-11114, WV-11533, WV-12503, WV-12504, WV-12505, WV-8553, WV-8555, WV-8556, WV-8557, WV-8582, WV-8583, WV-8584, WV-8585, WV-8586, WV-8587, WV-8588, WV-8589, WV-8590, WV-8591, WV-8592, WV-8593, WV-9058, WV-9059, WV-9060, WV-9061, WV-9696, WV-9697, and WV-9698.

[0387] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises one pattern of two different types of sugar modifications; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8097, WV-8098, WV-8099, WV-8100, WV-8101, WV-8102, and WV-8109.

[0388] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises one pattern of m and M; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8097, WV-8098, WV-8099, WV-8100, WV-8101, WV-8102, and WV-8109.

[0389] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mmMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8097, WV-8098, WV-8099, WV-8100, WV-8101, WV-8102, and WV-8109.

[0390] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises m and two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-12110, WV-12111, WV-12112, WV-12113, and WV-12114.

[0391] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMM; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-12110, WV-12111, WV-12112, WV-12113, and WV-12114.

[0392] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-13303, WV-13304, WV-13809, WV-14087, WV-14349, WV-14556, WV-14557, WV-14558, WV-14559, WV-14560, WV-14561, WV-14562, WV-14563, WV-14564, WV-14733, WV-14734, WV-14735, WV-14736, WV-14737, WV-14771, WV-15310, WV-15311, WV-15312, WV-15313, WV-15314, WV-15315, WV-15316, WV-15317, WV-15318, WV-15319, WV-15320, WV-15321 , WV-15351, WV-15352, WV-15353, WV-15354, WV-15355, WV-15356, WV-15357 , WV-15358, WV-15359, WV-15360, WV-15361, WV-15362, WV-15363, WV-1536 4, WV-15365, WV-15562, WV-15563, WV-15863, WV-15864 and WV-15887.

[0393] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8552, WV-8554, WV-8570, WV-8571, WV-8572, WV-8573, WV-8574, WV-8575, WV-8576, WV-8577, WV-8578, WV-8579, WV-8580, and WV-8581.

[0394] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-14552, WV-14553, WV-14554, and WV-14555.

[0395] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8456.

[0396] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8005, WV-8006, WV-8007, WV-8008, WV-8466, WV-8467, WV-8468, WV-8469, WV-8470, WV-8471, WV-8547, WV-8548, WV-8594, WV-13305, WV-13306, WV-13307, WV-13308, WV-13309, WV-13310, WV-13311, WV-13313, WV-13803, WV-13804, and WV-13805.

[0397] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8125.

[0398] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8314.

[0399] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9508, WV-9509, and WV-9510.

[0400] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9508, WV-9509, and WV-9510.

[0401] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8595, WV-8691, WV-8692, WV-8693, WV-8694, WV-8695, WV-8696, WV-9062, WV-9063, WV-9285, WV-9286, WV-9380, WV-9381, WV-9394, WV-9395, WV-9396, WV-9397, WV-9398, WV-9399, WV-9421, WV-9421, WV-9486, and WV-9487.

[0402] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9488, WV-9489, WV-9490, WV-9491, WV-9492, WV-9494, WV-9505, WV-9506, WV-9507, WV-8452, WV-8453, WV-8009, WV-8010, WV-8011, WV-8012, WV-8454, and WV-8455.

[0403] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8549, WV-8550, WV-8551, WV-8568, WV-8569, WV-13312, WV-14758, WV-14772, WV-15049, WV-15050, and WV-15051.

[0404] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a particular pattern of two different types of sugar modifications; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8121, WV-8129, WV-8118, WV-8126, WV-8472, WV-8473, WV-8474, WV-8475, and WV-8476.

[0405] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a particular pattern of two different types of sugar modifications; the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8119 and WV-8127.

[0406] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8115 and WV-8123.

[0407] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a particular pattern of two different types of sugar modifications; the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8120 and WV-8128.

[0408] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8116 and WV-8124.

[0409] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-14552, WV-14553, WV-14554, and WV-14555.

[0410] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8456.

[0411] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8005, WV-8006, WV-8007, WV-8008, WV-8466, WV-8467, WV-8468, WV-8469, WV-8470, WV-8471, WV-8547, WV-8548, WV-8594, WV-13305, WV-13306, WV-13307, WV-13308, WV-13309, WV-13310, WV-13311, WV-13313, WV-13803, WV-13804, and WV-13805.

[0412] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8125.

[0413] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8314.

[0414] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9508, WV-9509, and WV-9510.

[0415] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9508, WV-9509, and WV-9510.

[0416] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8595, WV-8691, WV-8692, WV-8693, WV-8694, WV-8695, WV-8696, WV-9062, WV-9063, WV-9285, WV-9286, WV-9380, WV-9381, WV-9394, WV-9395, WV-9396, WV-9397, WV-9398, WV-9399, WV-9421, WV-9421, WV-9486, and WV-9487.

[0417] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9488, WV-9489, WV-9490, WV-9491, WV-9492, WV-9494, WV-9505, WV-9506, and WV-9507.

[0418] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises a particular pattern of two different types of sugar modifications; the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8452 and WV-8453.

[0419] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8452 and WV-8453.

[0420] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8009, WV-8010, WV-8011, WV-8012, WV-8454, and WV-8455.

[0421] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8549, WV-8550, WV-8551, WV-8568, WV-8569, WV-13312, WV-14758, WV-14772, WV-15049, WV-15050, and WV-15051.

[0422] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMmm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8121 and WV-8129.

[0423] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two different types of sugar modifications and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8118, WV-8126, WV-8472, WV-8473, WV-8474, WV-8475, and WV-8476.

[0424] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMmm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8118, WV-8126, WV-8472, WV-8473, WV-8474, WV-8475, and WV-8476.

[0425] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMmmm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8119 and WV-8127.

[0426] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMmmm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8115 and WV-8123.

[0427] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mmMmm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8120 and WV-8128.

[0428] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mmMmm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8116 and WV-8124.

[0429] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, and WV-8664.

[0430] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, and WV-8664.

[0431] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8645, WV-8646, WV-8647, WV-8648, WV-8661, WV-8662, WV-8663, and WV-8664.

[0432] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive M's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8637, WV-8638, WV-8639, WV-8640, WV-8653, WV-8654, WV-8655, WV-8656, WV-8665, WV-8666, WV-8667, WV-8668, WV-8669, WV-8670, WV-8671, WV-8672, WV-12947, WV-12948, WV-12949, WV-12950, ​​WV-12951, WV-12952, and WV-12953. , WV-12954, WV-12955, WV-12956, WV-12957, WV-12958, WV-12959, WV-12960, WV-12961, WV-12962, WV-12963, WV-12964, WV-1296 5, WV-12966, WV-12967, WV-12968, WV-12969, WV-12970, WV-12971, WV-12972, WV-12973, WV-12974, WV-12975 and WV-12976.

[0433] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises two or more consecutive m's; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-12977, WV-12978, WV-12979, WV-12980, WV-12981, WV-12982, WV-12983, WV-12984, WV-12985, WV-12986, WV-12987, WV-12988, WV-12989, WV-12990, WV-12991, Examples include WV-12992, WV-12993, WV-12994, WV-12995, WV-12996, WV-12997, WV-12998, WV-12999, WV-13000, WV-13001, WV-13002, WV-13003, WV-13004, WV-13005, WV-13006, WV-13007 and WV-13008.

[0434] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMM; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9887, WV-9888, WV-10245, and WV-10246.

[0435] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMM; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9871 and WV-9872.

[0436] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMMMM; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. A non-limiting example of such an oligonucleotide is WV-12101.

[0437] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMM; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9873 and WV-9874.

[0438] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMM; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9885, WV-9886, WV-10243, and WV-10244.

[0439] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-9526, WV-9527, WV-9528, WV-9529, WV-9530, WV-9531, WV-9532, WV-9533, WV-9590, WV-9591, WV-9592, and WV-9593.

[0440] In some embodiments of oligonucleotides having an asymmetric format, the first wing comprises mMMMm; and the second wing comprises a different sugar modification or a different combination or pattern of sugar modifications. Non-limiting examples of such oligonucleotides include WV-8610, WV-8611, WV-8612, WV-8613, WV-8614, WV-8615, WV-8616, WV-8617, WV-8618, WV-8619, WV-8629, WV-8632, WV-8673, WV-8674, WV-8675, WV-8676, WV-8677, WV-8678, WV-8679, WV-8680, WV-8681, WV-8682, WV-8683, WV-8684, WV-8685, WV-8686, WV-8687, and WV-8688.

[0441] In some embodiments, two or more consecutive m's are mm. In some embodiments, two or more consecutive m's are mmm. In some embodiments, two or more consecutive m's are mmmm. In some embodiments, two or more consecutive m's are mmmmmm. In some embodiments, two or more consecutive m's are mmmmmm. In some embodiments, two or more consecutive m's are mmmmmm. In some embodiments, two or more consecutive m's are mmmmmmm.

[0442] In some embodiments, two or more consecutive M's are MM. In some embodiments, two or more consecutive M's are MMM. In some embodiments, two or more consecutive M's are MMMM. In some embodiments, two or more consecutive M's are MMMMM. In some embodiments, two or more consecutive M's are MMMMMM. In some embodiments, two or more consecutive M's are MMMMMMM.

[0443] In some embodiments, two or more consecutive F's are FF. In some embodiments, two or more consecutive F's are FFF. In some embodiments, two or more consecutive F's are FFFF. In some embodiments, two or more consecutive F's are FFFFF. In some embodiments, two or more consecutive F's are FFFFFF. In some embodiments, two or more consecutive F's are FFFFFFF. In some embodiments, two or more consecutive F's are FFFFFFF.

[0444] In some embodiments, the oligonucleotides of the present disclosure comprise sugar modifications. In some embodiments, the oligonucleotides can comprise any sugar described herein or known in the art. In some embodiments, the first wing of an oligonucleotide having an asymmetric format can comprise any sugar described herein or known in the art, and the second wing of the oligonucleotide does not comprise a sugar. In some embodiments, the first wing of an oligonucleotide having an asymmetric format can comprise any sugar described herein or known in the art, and the second wing of the oligonucleotide comprises a different sugar.

[0445] In some embodiments, the sugar is: It has the structure TIFF2026016363000013.tif1473.

[0446] Modified sugars can be incorporated into provided oligonucleotides. In some embodiments, the modified sugar comprises one or more substituents at the 2' position including one of the following: -F; -CF, -CN, -N, -NO, -NO, -OR', -SR', or -N(R'), where each R' is independently as described in this disclosure; -O-(C1-C 10 alkyl), -S-(C1-C 10 alkyl), -NH-(C1-C 10 alkyl) or -N(C1-C 10 alkyl)2; -O-(C2-C 10 alkenyl), -S-(C2-C 10 alkenyl), -NH-(C2-C 10 alkenyl) or -N(C2-C 10 alkenyl)2; -O-(C2-C 10 alkynyl), -S-(C2-C 10 alkynyl), -NH-(C2-C 10 alkynyl) or -N(C2-C 10 alkynyl)2; or -O-(C1-C 10 alkylene)-O-(C1-C 10 alkyl), -O-(C1-C 10 Alkylene)-NH-(C1-C 10 alkyl) or -O-(C1-C 10 Alkylene)-NH-(C1-C 10 alkyl)2, -NH-(C1-C 10 alkylene)-O-(C1-C 10 alkyl) or -N(C1-C 10 Alkyl)-(C1-C 10 alkylene)-O-(C1-C 10 alkyl), where alkyl, alkylene, alkenyl, and alkynyl can be substituted or unsubstituted. Examples of substituents include, but are not limited to, —O(CH) n OCH3 and -O(CH2) nExamples of modified sugars include NH2 (where n is 1 to about 10), MOE, DMAOE, and DMAEOE. Also contemplated herein are the modified sugars described in WO 2001 / 088198; and Martin et al., Helv. Chim. Acta, 1995, 78, 486-504. In some embodiments, the modified sugar comprises one or more groups selected from the following: a substituted silyl group, an RNA cleaving group, a reporter group, a fluorescent label, an intercalating agent, a group for improving the pharmacokinetic properties of nucleic acids, a group for improving the pharmacodynamic properties of nucleic acids, or other substituents with similar properties. In some embodiments, the modification is made at one or more of the 2', 3', 4', 5', or 6' positions of the sugar or modified sugar, including the 3' position of the sugar on the 3'-terminal nucleotide or the 5' position of the sugar on the 5'-terminal nucleotide.

[0447] In some embodiments, the 2'-modification is 2'-F.

[0448] In some embodiments, the 2'-OH of the ribose is substituted with a substituent, including one of the following: -H, -F; -CF, -CN, -N, -NO, -NO, -OR', -SR', or -N(R'), where each R' is independently as described in this disclosure; -O-(C-C 10 alkyl), -S-(C1-C 10 alkyl), -NH-(C1-C 10 alkyl) or -N(C1-C 10 alkyl)2; -O-(C2-C 10 alkenyl), -S-(C2-C 10 alkenyl), -NH-(C2-C 10 alkenyl) or -N(C2-C 10 alkenyl)2; -O-(C2-C 10 alkynyl), -S-(C2-C 10 alkynyl), -NH-(C2-C 10 alkynyl) or -N(C2-C 10 alkynyl)2; or -O-(C1-C 10 alkylene)-O-(C1-C 10 alkyl), -O-(C1-C 10 Alkylene)-NH-(C1-C10 alkyl) or -O-(C1-C 10 Alkylene)-NH-(C1-C 10 alkyl)2, -NH-(C1-C 10 alkylene)-O-(C1-C 10 alkyl) or -N(C1-C 10 Alkyl)-(C1-C 10 alkylene)-O-(C1-C 10 In some embodiments, the 2'-OH is substituted with -H (deoxyribose). In some embodiments, the 2'-OH is substituted with -F. In some embodiments, the 2'-OH is substituted with -OR'. In some embodiments, the 2'-OH is substituted with -OMe. In some embodiments, the 2'-OH is substituted with -OCHCHOMe.

[0449] Modified sugars also include locked nucleic acids (LNAs). In some embodiments, two substituents on a sugar carbon atom together form a divalent moiety. In some embodiments, the two substituents are on two different sugar carbon atoms. In some embodiments, the divalent moiety formed has the structure -L-, as defined herein. In some embodiments, -L- is -O-CH2-, where -CH2- is optionally substituted. In some embodiments, -L- is -O-CH2-. In some embodiments, -L- is -O-CH(Et)-. In some embodiments, -L- is between C2 and C4 of the sugar moiety. In some embodiments, locked nucleic acids have the structure shown below. A locked nucleic acid of the following structure is shown, where B represents a nucleobase or modified nucleobase as described herein, e.g., R 2S and R 4S is R, which together with the intervening atoms forms a ring. In some embodiments, the modified nucleoside is: TIFF2026016363000014.tif1573 (wherein B is a base) It has the following structure.

[0450] In some embodiments, the modified sugar is an ENA, such as those described in Seth et al., J Am Chem Soc. 2010 October 27;132(42):14942-14950. In some embodiments, the modified sugar is one found in an XNA (xenonucleic acid), such as arabinose, anhydroxylitol, threose, 2'fluoroarabinose, or cyclohexene.

[0451] Modified sugars include cyclobutyl or cyclopentyl moieties in place of the pentofuranosyl sugar. Representative U.S. patents that teach the preparation of such modified sugars include, but are not limited to, U.S. Pat. Nos. 4,981,957; 5,118,800; 5,319,080; and 5,359,044. Some contemplated modified sugars include those in which the oxygen atom in the ribose ring has been replaced with nitrogen, sulfur, selenium, or carbon. In some embodiments, the modified sugar is a modified ribose in which the oxygen atom in the ribose ring has been replaced with nitrogen, which is optionally substituted with an alkyl group (e.g., methyl, ethyl, isopropyl, etc.).

[0452] Non-limiting examples of modified sugars include glycerol, which forms glycerol nucleic acid (GNA). An example of GNA is shown below and is described in Zhang, R et al., J. Am. Chem. Soc., 2008, 130, 5846-5847; Zhang L, et al., J. Am. Chem. Soc., 2005, 127, 4174-4175 and Tsai CH et al., PNAS, 2007, 14598-14603. In some embodiments, the nucleoside is: TIFF2026016363000015.tif1673 (wherein B is a base) It has the following structure.

[0453] Flexible nucleic acids (FNAs) based on mixed acetal aminals of formyl glycerol are described in Joyce GF et al., PNAS, 1987, 84, 4398-4402 and Heuberger BD and Switzer C, J. Am. Chem. Soc., 2008, 130, 412-413. In some embodiments, the nucleoside is as follows: TIFF2026016363000016.tif1673(where B is a base) has the structure of.

[0454] Further non-limiting examples of modified sugars and / or modified nucleosides and / or modified nucleotides include hexopyranosyl (6'-to-4'), pentopyranosyl (4'-to-2'), pentopyranosyl (4'-to-3'), 5'-deoxy-5'-C-malonyl, squaryldiamide, and tetrofuranosyl (3'-to-2') sugars. In some embodiments, the modified nucleoside comprises a hexopyranosyl (6'-to-4') sugar and has the following formula: TIFF2026016363000017.tif3573(where X S corresponds to the P-modifying group "-XLR 1 " described herein, where XLR 1 is equivalent to X-L-R 1 and X, L, and R 1 are as defined in formula I disclosed herein, and B is a base) has any one of the structures of.

[0455] In some embodiments, the modified nucleotide comprises a pentopyranosyl (4'-to-2') sugar and has the following formula: TIFF2026016363000018.tif1473(where X S corresponds to the P-modifying group "-XLR 1 " described herein, where XLR 1 is equivalent to X-L-R 1 and X, L, and R 1 are as defined in formula I disclosed herein, and B is a base) has any one of the following structures.

[0456] In some embodiments, the modified nucleotide comprises a pentopyranosyl (4’ to 3’) sugar and has the following formula: TIFF2026016363000019.tif1673 (where X S corresponds to the P-modifying group “-XLR 1 ” described herein, where XLR 1 is equivalent to X-L-R 1 and X, L, and R 1 are as defined in Formula I disclosed herein, and B is a base). is any one of the following.

[0457] In some embodiments, the modified nucleotide comprises a tetrofuranosyl (3’ to 2’) sugar and has the following formula: TIFF2026016363000020.tif1573 (where X S corresponds to the P-modifying group “-XLR 1 ” described herein, where XLR 1 is equivalent to X-L-R 1 and X, L, and R 1 are as defined in Formula I disclosed herein, and B is a base). is any of the following.

[0458] In some embodiments, the modified nucleotide comprises a modified sugar and has the following formula: TIFF2026016363000021.tif2473 (where X S corresponds to the P-modifying group “-XLR 1 ” described herein, where XLR 1 is equivalent to X-L-R 1 and X, L, and R 1 are as defined in Formula I disclosed herein, and B is a base). is any one of the following.

[0459] In some embodiments, one or more hydroxyl groups of the sugar moiety are optionally independently substituted with a halogen, R', -N(R')2, -OR', or -SR', where each R' is independently described in this disclosure.

[0460] In some embodiments, the modified nucleotide is as shown below, where X S is a P-modifying group "-XLR" as described herein. 1 ", where XLR 1 is XLR 1 X, L and R are equivalent to 1 is as defined in Formula I disclosed herein, B is a base, and X 1 -S-, -Se-, -CH2-, -NMe-, -NEt- and -NiPr- Selected from TIFF2026016363000022.tif7173.

[0461] In some embodiments, at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 12 %, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more) (inclusive) are qualified. In some embodiments, only purine residues are modified (e.g., about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 9%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more [e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more] would qualify). In some embodiments, only pyrimidine residues are modified (e.g., about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, or ...3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, or about 1%, 2%, 2%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, 3%, , 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more [e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more] are modified). In some embodiments, both purines and pyrimidines are modified.

[0462] Modified sugars can be prepared by methods known in the art, including, but not limited to, the following: A. Eschenmoser, Science (1999), 284:2118; M. Bohringer et al., Helv. Chim. Acta (1992), 75:1416-1477; M. Egli et al., J. Am. Chem. Soc. (2006), 128(33):10847-56; A. Eschenmoser in Chemical Synthesis: Gnosis to Prognosis, C. Chatgilialoglu and V. Sniekus, Ed., (Kluwer Academic, Netherlands, 1996), p. 293; K.-U. Schoning et al., Science (2000), 290:1347-1351; A. Eschenmoser et al. al, Helv. Chim. Acta (1992), 75:218; J. Hunziker et al, Helv. Chim. Acta (1993), 76:259; G. Otting et al, Helv. Chim. Acta (1993), 76:2701; K. Groebke et al, Helv. Chim. Acta (1998), 81:375; and A. Eschenmoser, Science (1999), 284:2118. Modifications for 2' modifications can be found in Verma, S. et al. Annu. Rev. Biochem. 1998, 67, 99-134 and all references therein. Specific modifications to ribose can be found in the following references: 2'-fluoro (Kawasaki et al., J. Med. Chem., 1993, 36, 831-841), 2'-MOE (Martin, P. Helv. Chim. Acta 1996, 79, 1930-1938), "LNA" (Wengel, J. Acc. Chem. Res. 1999, 32, 301-310). In some embodiments, the modified sugar is any of those described in WO 2012 / 030683, which is incorporated by reference and / or depicted herein. In some embodiments, the modified sugar is any modified sugar described in any of the following references: Gryaznov, S; Chen, J.-KJAm.Chem.Soc.1994,116,3143;Hendrix et al.1997 Chem.Eur.J.3:110;Hyrup et al.1996 Bioorg.Med.Chem.4:5;Jepsen et al.2004 Oligo.14:130-146;Jones et al.J.Org.Chem.1993,58,2983;Koizumi et al.2003 Nuc.Acids Res.12:3267-3273;Koshkin et al.1998 Tetrahedron 54:3607-3630;Kumar et al.1998 Bioo.Med.Chem.Let.8:2219-2222;Lauritsen et al.2002 Chem.Comm.5:530-531;Lauritsen et al. al.2003 Bioo.Med.Chem.Lett.13:253-256;Mesmaeker et al.Angew.Chem.,Int.Ed.Engl.1994,33,226;Morita et al.2001 Nucl.Acids Res.Supp.1:241-242;Morita et al.2002 Bioo.Med.Chem.Lett.12:73-76;Morita et al.2003 Bioo.Med.Chem.Lett.2211-2226;Nielsen et al.1997 Chem.Soc.Rev.73;Nielsen et al.1997 J.Chem.Soc.Perkins Transl.1:3423-3433;Obika et al.1997 Tetrahedron Lett.38(50):8735-8;Obika et al.1998 Tetrahedron Lett.39:5401-5404;Pallan et al.2012 Chem.Comm.48:8195-8197;Petersen et al.2003 TRENDS Biotech.21:74-81;Rajwanshi et al.1999 Chem.Commun.1395-1396;Schultz et al.1996 Nucleic Acids Res.24:2966;Seth et al.2009 J.Med.Chem.52:10-13;Seth et al.2010 J.Med.Chem.53:8309-8318; Seth et al.2010 J.Org.Chem.75:1569-1581; Seth et al.2012 Bioo.Med.Chem.Lett.22:296-299; Seth et al.2012 Mol.Ther-Nuc.Acids.1,e47; Seth,Punit P; Siwkowski,Andrew; Allerson,Charles R; Vasquez,Guillermo; Lee,Sam; Prakash,Thazha P; Kinberger,Garth; Migawa,Michael T; Gaus,Hans; Bhat,Balkrishen; et al.From Nucleic Acids Symposium Series(2008),52(1),553-554; Singh et al.1998 Chem.Comm.1247-1248; Singh et al.1998 J.Org.Chem.63:10035-39; Singh et al.1998 J.Org.Chem.63:6078-6079; Sorensen 2003 Chem.Comm.2130-2131; Ts’o et al.Ann.N.Y.Acad.Sci.1988,507,220; Van Aerschot et al.1995 Angew.Chem.Int.Ed.Engl.34:1338; Vasseur et al.J.Am.Chem.Soc.1992,114,4006; International Publication No. 20070900071 pamphlet; the same No. 20070900071 pamphlet; or the same No. 2016 / 079181 pamphlet.

[0463] In some embodiments, the modified sugar moiety is an optionally substituted pentose or hexose moiety. In some embodiments, the modified sugar moiety is an optionally substituted pentose. In some embodiments, the modified sugar moiety is an optionally substituted hexose moiety. In some embodiments, the modified sugar moiety is an optionally substituted ribose or hexitol moiety. In some embodiments, the modified sugar moiety is an optionally substituted ribose moiety. In some embodiments, the modified sugar moiety is an optionally substituted hexitol moiety.

[0464] In some embodiments, an example of a modified nucleotide is selected from the following: TIFF2026016363000023.tif7273 In some embodiments, the nucleotide is: TIFF2026016363000024.tif4173. In some embodiments, the nucleotide is: TIFF2026016363000025.tif1473 (in the formula, R 1 and R 2 are independently -H, -F, -OMe, -MOE, or substituted or unsubstituted C 1~6 alkyl); TIFF2026016363000026.tif2373(in the formula, R e is a substituted or unsubstituted C 1~6 alkyl or H), TIFF2026016363000027.tif8573. Other chemically modified sugars are described in WO 2008 / 101157, WO 2007 / 134181, WO 2016 / 167780, and U.S. Patent Application Publication No. 2005-0130923. In some embodiments, the nucleotide and flanking nucleoside have a structure selected from the following: It has the structure TIFF2026016363000028.tif2073.

[0465] In some embodiments, a locked nucleic acid, i.e., an LNA or LNA nucleoside or LNA nucleotide, is or comprises a nucleic acid monomer having a bridge connecting two carbon atoms between the 4' and 2' positions of the nucleoside sugar unit, thereby forming a bicyclic sugar. Examples of such bicyclic sugars include, but are not limited to, α-L-methyleneoxy (4'-CH2-O-2')LNA, β-D-methyleneoxy (4'-CH2-O-2')LNA, ethyleneoxy (4'-(CH2)2-O-2')LNA, aminooxy (4'-CH2-ON(R)-2')LNA, and oxyamino (4'-CH2-N(R)-O-2')LNA. In some embodiments, R is R1 or R2.

[0466] Examples of nucleosides having modified sugar moieties include, but are not limited to, 5'-vinyl, 5'-methyl (R or S), 4'-S, 2'-F, 2'-OCH, 2'-OCHCH, 2'-OCHCHF, and 2'-O(CH). 20 Nucleosides containing a CH3 substituent are also included. The substituent at the 2' position can be allyl, amino, azido, thio, O-allyl, O-C1-C 10 Alkyl, OCF3, OCH2F, O(CH2)2SCH3, O(CH2)2-ON(R m )(R n ), O-CH2-C(=O)-N(R m )(R n ) and O-CH2-C(=O)-N(R1)-(CH2)2-N(R m )(R n ) can be selected from R1, R m and R n are independently H or substituted or unsubstituted C1-C 10 It is alkyl.

[0467] In some embodiments, bicyclic nucleosides include any modified nucleoside containing a bicyclic sugar moiety. Examples of bicyclic nucleic acids (BNAs) include, but are not limited to, nucleosides containing a bridge between the 4' and 2' ribosyl ring atoms. In some embodiments, the antisense compounds provided herein comprise one or more BNA nucleosides, wherein the bridge has the formula: 4'-(CH2)-O-2' (LNA); 4'-(CH2)-S-2'; 4,-(CH2)2-O-2' (ENA); 4'-CH(CH3)-O-2' and 4'-CH(CHOCH3)-O-2' (and analogs thereof; see U.S. Pat. No. 7,399,845); 4'-C(CH3)(CH3)-O-2' (and analogs thereof; see WO 2009 / 010162). See PCT / US2008 / 068922, published as WO 06478; 4'-CH2-N(OCH3)-2' (and analogs thereof; see PCT / US2008 / 064591, published as WO 2008 / 150729); 4'-CH2-ON(CH3)-2' (see U.S. Patent Application Publication No. 2004-0171570); 4'-CH2-N(R)-O-2' (where R is H, C1-C 12 alkyl or a protecting group) (see U.S. Pat. No. 7,427,672); 4'-CH2-C(H)(CH3)-2' (see Chattopadhyaya et al, J. Org. Chem., 2009, 74, 118-134); and 4,-CH2-C(=CH2)-2' (and analogs thereof; see PCT / U.S. Patent Application Publication No. 2008 / 066154, published as WO 2008 / 154401).

[0468] Further nucleosides have been reported in the literature (see, for example, Srivastava et al, J. Am. Chem. Soc., 2007, 129(26) 8362-8379; Frieden et al, Nucleic Acids Research, 2003, 21, 6365-6372; Elayadi et al, Curr. Opinion Inverts. Drugs, 2001, 2, 558-561; Braasch et al, Chem. Biol, 2001, 8, 1-7; Oram et al, Curr. Opinion Mol Ther., 2001, 3, 239-243; Wahlestedt et al, Proc. Natl Acad. Sci. USA, 2000, 97, 5633-5638; Singh et al. al,Chem.Commun.,1998,4,455-456;Koshkin et al,Tetrahedron,1998,54,3607-3630;Kumar et al,Bioorg.Med.Chem.Lett.,1998,8,2219-2222;Singh et al. al, J. Org. Chem., 1998, 63, 10035-10039; U.S. Patent Nos. 7,399,845; 7,053,207; 7,034,133; 6,794,499; 6,770,748; 6,670,461; 6,525,191; 6,268,490; U.S. Patent Application Publication Nos. 2008-0039618; 2007-0287831; 2004-0171570; U.S. Patent Application No. 12 / 129,154; Nos. 61 / 099,844; 61 / 097,787; 61 / 086,231; 61 / 056,564; 61 / 026,998; 61 / 026,995; 60 / 989,574; WO 2007 / 134181; WO 2005 / 021570; WO 2004 / 106356; and PCT International Application Nos. PCT / US 2008 / 068922; WO 2008 / 066154; and WO 2008 / 064591).

[0469] In some embodiments, bicyclic nucleosides can be prepared with one or more stereochemical sugar configurations, including, for example, α-L-ribofuranose and β-D-ribofuranose (see PCT International Application No. PCT / Danish Patent No. 98 / 00393, published as WO 99 / 14226). In some embodiments, monocyclic nucleosides are nucleosides containing modified sugar moieties that are not bicyclic sugars. In some embodiments, the sugar moiety or sugar moiety analog of the nucleoside can be modified or substituted at any position. In some embodiments, 4'-2' bicyclic nucleosides, i.e., 4' to 2' bicyclic nucleosides, are bicyclic nucleosides containing a furanose containing a bridge connecting two carbon atoms of the furanose ring, which connects the 2' and 4' carbon atoms of the sugar ring. In some embodiments, the bicyclic sugar moiety of a BNA nucleoside includes, but is not limited to, compounds having at least one bridge between the 4' and 2' carbon atoms of the pentofuranosyl sugar moiety, including, but not limited to, the following: -[C(R a )(R b )] n -, -C(R a )=C(R b )-, -C(R a )=N-, -C(=NR a )-, -C(=O)-, -C(=S)-, -O-, -Si(R a )2-, -S(=O)x- and -N(R a )—(wherein x is 0, 1, or 2; n is 1, 2, 3, or 4; R a and R b are each independently H, a protecting group, hydroxyl, C1-C 12 Alkyl, substituted C1-C 12 Alkyl, C2-C 12 Alkenyl, substituted C2-C 12 Alkenyl, C2-C 12 Alkynyl, substituted C2-C 12 Alkynyl, C5-C 20 Aryl, substituted C5-C 20aryl, heterocyclyl radical, substituted heterocyclyl radical, heteroaryl, substituted heteroaryl, C5-C7 alicyclic radical, substituted C5-C7 alicyclic radical, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)2-J1), or sulfoxyl (S(=O)-J1); each J1 and J2 is independently H, C1-C 12 Alkyl, substituted C1-C 12 Alkyl, C2-C 12 Alkenyl, substituted C2-C 12 Alkenyl, C2-C 12 Alkynyl, substituted C2-C 12 Alkynyl, C5-C 20 Aryl, substituted C5-C 20 Aryl, acyl (C(=O)-H), substituted acyl, heterocyclic radical, substituted heterocyclic radical, C1-C 12 Aminoalkyl, substituted C1-C 12 and a moiety containing a bridge containing one or 1 to 4 linking groups independently selected from the group consisting of aryl, arylamino, arylalkyl ...

[0470] In some embodiments, the bridge of the bicyclic sugar moiety is —[C(R a )(R b )] n -, -[C(R a )(R b )] n -O-, -C(R a R b )-N(R)-O- or -C(R a R b In some embodiments, the bridges are 4'-CH2-2', 4'-(CH2)2-2', 4'-(CH2)3-2', 4'-CH2-O-2', 4'-(CH2)2-O-2', 4'-CH2-ON(R)-2', and 4'-CH2-N(R)-O-2'-, where each R is independently H, a protecting group, or a C1-C 12 It is alkyl.

[0471] In some embodiments, bicyclic nucleosides are further distinguished by their isomeric conformation. For example, nucleosides containing a 4'-(CH2)-O-2' bridge can be in the α-L- or β-D-conformation. α-L-methyleneoxy(4'-CH2-O-2') BNAs have been incorporated into antisense oligonucleotides that exhibit antisense activity (Frieden et al., Nucleic Acids Research, 2003, 21, 6365-6372).

[0472] In some embodiments, bicyclic nucleosides include those with a 4' to 2' bridge, where such bridges include, but are not limited to, aL-4'-(CH2)-O-2', PD-4'-CH2-O-2', 4'-(CH2)2-O-2', 4'-CH2-ON(R)-2', 4'-CH2-N(R)-O-2', 4'-CH(CH3)-O-2', 4'-CH2-S-2', 4'-CH2-N(R)-2', 4'-CH2-CH(CH3)-2', and 4'-(CH2)3-2', where R is H, a protecting group, or a C1-C 12 It is alkyl.

[0473] Various bicyclic nucleotide analogs with 4'-to-2' bridging groups, such as 4'-CH2-O-2' and 4'-CH2-S-2', have also been prepared (Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222). The preparation of oligodeoxyribonucleotide duplex strands containing bicyclic nucleosides for use as substrates for nucleic acid polymerases has also been described (Wengel et al., WO 99 / 14226). Furthermore, the synthesis of 2'-amino-BNAs, a novel conformationally restricted, high-affinity oligonucleotide analog, has been described in the art (Singh et al., J. Org. Chem., 1998, 63, 10035-10039). Furthermore, 2'-amino- and 2'-methylamino-BNAs have been prepared, and the thermal stability of their duplex strands with complementary RNA and DNA strands has been reported.

[0474] One carbocyclic bicyclic nucleoside with a 4'-(CH2)3-2' bridge and an alkenyl analog bridged 4'-CH=CH-CH2-2' have been described (Frier et al., Nucleic Acids Research, 1997, 25(22), 4429-4443 and Albaek et al., J. Org. Chem., 2006, 71, 7731-7740). The synthesis and preparation of carbocyclic bicyclic nucleosides have also been described, along with oligomerization and biochemical studies (Srivastava et al., J. Am. Chem. Soc. 2007, 129(26), 8362-8379).

[0475] In some embodiments, bicyclic nucleosides include, but are not limited to, α-L-methyleneoxy (4'-CH2-O-2') BNA, β-D-methyleneoxy (4'-CH2-O-2') BNA, ethyleneoxy (4'-(CH2)2-O-2') BNA, aminooxy (4'-CH2-ON(R)-2') BNA, oxyamino (4'-CH2-N(R)-O-2') BNA, methyl These include methyl(methyleneoxy) (4'-CH(CH3)-O-2') BNA (also called restricted ethyl or cEt), methylenethio (4'-(CH2)-S-2') BNA, methylene-amino (4'-CH2-N(R)-2') BNA, methyl carbocycle (4'-CH2-CH(CH3)-2') BNA, propylene carbocycle (4'-(CH2)3-2') BNA, and vinyl BNA.

[0476] In some embodiments, LNA compounds include, but are not limited to, compounds having at least one bridge between the 4' and 2' sugar positions, where each bridge is independently: -[C(R1)(R2)] n -, -C(R1)=(R2)-, -C(R1)=N-, -C(NR1)=N-, -C(=O)-, -C(=S)-, -O-, -Si(R1)2-, -S(=O) x - and -N(R1)- (wherein x is 0, 1, or 2; n is 1, 2, 3, or 4; each of R1 and R2 is independently H, a protecting group, hydroxyl, C1-C 12 Alkyl, substituted C1-C12 Alkyl, C2-C 12 Alkenyl, substituted C2-C 12 Alkenyl, C2-C 12 Alkynyl, substituted C2-C 12 Alkynyl, C5-C 20 Aryl, substituted C5-C 20 aryl, heterocyclic radical, substituted heterocyclic radical, heteroaryl, substituted heteroaryl, C5-C7 alicyclic radical, substituted C5-C7 alicyclic radical, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)2-J1), or sulfoxyl (S(=O)-J1); each J1 and J2 is independently H, C1-C 12 Alkyl, substituted C1-C 12 Alkyl, C2-C 12 Alkenyl, substituted C2-C 12 Alkenyl, C2-C 12 Alkynyl, substituted C2-C 12 Alkynyl, C5-C 20 Aryl, substituted C5-C 20 Aryl, acyl (C(=O)-H), substituted acyl, heterocyclic radical, substituted heterocyclic radical, C1-C 12 Aminoalkyl, substituted C1-C 12 A bridge containing one or two to four linking groups independently selected from the group consisting of: -C(R1)(R2), -C(R3), -C(R4), -C(R5), -C(R6), -C(R7), -C(R8), -C(R9), -C(R10), -C(R11), -C(R12), -C(R13), -C(R14), -C(R15), -C(R26), -C(R27), -C(R28), -C(R29 ... n -, -[C(R1)(R2)] n -O-, -C(R1R2)-N(R1)-O-, or -C(R1R2)-ON(R1)-. Further, other bridges encompassed within the definition of LNA are 4'-CH2-2', 4'-(CH2)2-2', 4'-(CH2)3-2', 4'-CH2-O-2', 4'-(CH2)2-O-2', 4'-CH2-ON(R1)-2', and 4'-CH2-N(R1)-O-2' bridges, where R1 and R2 are independently H, a protecting group, or a C1-C 12The term "LNA" is an alkyl. Also included within the definition of LNA are LNAs in which the 2'-hydroxyl group of the ribosyl sugar ring is linked to the 4' carbon atom of the sugar ring, thereby forming a methyleneoxy (4'-CH2-O-2') bridge, thereby forming a bicyclic sugar moiety. The bridge can be a methylene (-CH2-) group connecting the 2' oxygen atom and the 4' carbon atom, for which the term methyleneoxy (4'-CH2-O-2') LNA is used. In some embodiments, when the bicyclic sugar moiety has an ethylene bridging group at this position, the term ethyleneoxy (4'-CH2CH2-O-2') LNA is used. The isomer of methyleneoxy (4'-CH2-O-2') LNA, α-L-methyleneoxy (4'-CH2-O-2'), is also included within the definition of LNA used herein.

[0477] In some embodiments, the 2'-modification is -F. In some embodiments, the 2'-modification is FANA. In some embodiments, the 2'-modification is FRNA.

[0478] In some embodiments, the sugar modification is a 5'-modification, such as R-5'-Me, S-5'-Me, and the like.

[0479] In some embodiments, the sugar modification changes the size of the sugar ring. In some embodiments, the sugar modification is the sugar moiety of the FHNA.

[0480] In some embodiments, sugar modifications replace the sugar moiety with another cyclic or acyclic moiety, examples of which are known in the art and include, but are not limited to, those used in morpholinos (optionally with their phosphorodiamidate linkages), glycol nucleic acids, etc.

[0481] In some embodiments, modified tetrahydropyran nucleosides or modified THP nucleosides are nucleosides with a six-membered tetrahydropyran "sugar" replacing the pentofuranosyl residue in a normal nucleoside, and can be referred to as sugar surrogates. Modified THP nucleosides include, but are not limited to, those referred to in the art as hexitol nucleic acids (HNA), anitol nucleic acids (ANA), and mannitol nucleic acids (MNA) (see Leumann, Bioorg. Med. Chem., 2002, 10, 841-854), as well as fluoro-HNA (F-HNA), which has a tetrahydropyranyl ring system, as shown below:

[0482] In some embodiments, the sugar surrogate comprises a ring having six or more atoms and two or more heteroatoms. For example, their use in nucleosides and oligomeric compounds containing morpholino sugar moieties has been reported (see, e.g., Braasch et al., Biochemistry, 2002, 41, 4503-4510; and U.S. Pat. Nos. 5,698,685; 5,166,315; 5,185,444; and 5,034,506).

[0483] Combinations of modifications are also provided, including, but not limited to, 2'-F-5'-methyl substituted nucleosides (see WO 2008 / 101157 for other disclosed 5',2'-bis substituted nucleosides) and replacement of the ribosyl ring oxygen atom with S and another substitution at the 2'-position (see U.S. Patent Application Publication No. 2005-0130923) or alternatively 5'-substitution of bicyclic nucleic acids (see WO 2007 / 134181) where the 4'-CH2-O-2' bicyclic nucleoside is further substituted at the 5'-position with a 5'-methyl or 5'-vinyl group. The synthesis and preparation...

Claims

1. In the oligonucleotide, (Op)n(Sp)m (In the formula, Sp indicates the S configuration of the chiral linking phosphorus of the chiral modified internucleotide linkage; Op represents the achiral linking phosphorus of the natural phosphate bond, and each of n and m is independently 1 to 20; and each wing independently comprises one or more nucleobases An oligonucleotide characterized in that it contains a pattern of backbone chiral centers (linked phosphorus) of:

2. 2. The oligonucleotide of claim 1, wherein the pattern of backbone chiral centers comprises (Sp)t[(Op / Rp)n(Sp)m]y, where t is 1 to 20.

3. 2. The oligonucleotide of claim 1, wherein the pattern of backbone chiral centers comprises (Np)t[(Op / Rp)n(Sp)m]y, where t is 1 to 20, Np is Sp or Rp, and Rp indicates the S configuration of the chiral linkage phosphorus of the chiral modified internucleotide linkage.

4. The oligonucleotide according to any one of claims 1 to 3, wherein n is 1.

5. The oligonucleotide according to claim 4, wherein m is 2 or more.

6. The oligonucleotide according to claim 5, wherein t is 2 or more.

7. 7. The oligonucleotide according to claim 6, comprising a wing-core-wing structure.

8. A composition comprising an oligonucleotide, wherein the oligonucleotide comprises a first wing, a second wing, and a core in a format of first wing-core-second wing or second wing-core-first wing, and wherein the first wing, the second wing, and the core each comprise different sugars or sugar modifications, or combinations or patterns thereof, and / or internucleotide linkages, or combinations or patterns thereof, and / or internucleotide linkage stereochemistry, or combinations or patterns thereof.

9. 9. The composition of claim 8, wherein the oligonucleotide has the ability to reduce the level, expression and / or activity of a target gene or its gene product.

10. 10. The composition of claim 8 or 9, wherein the oligonucleotide has the ability to reduce the level, expression and / or activity of a target gene or its gene product via a mechanism involving RNase H.

11. 11. The composition of any one of claims 8 to 10, wherein the oligonucleotide has the ability to reduce the level, expression and / or activity of a target gene or its gene product via a mechanism involving steric hindrance.

12. 12. The composition of claim 8, wherein the oligonucleotide is capable of annealing to a target mRNA and reducing the translation level of the target mRNA.

13. 13. The composition of any one of claims 8 to 12, wherein the oligonucleotide has the ability to anneal to a target RNA and reduce the translation level of the target RNA via a mechanism involving steric hindrance.

14. 14. The composition of any one of claims 8 to 13, wherein the core is capable of annealing to a target nucleic acid, forming a substrate for RNase H, and enabling cleavage of the target nucleic acid by RNase H.

15. 15. The composition of any one of claims 8 to 14, wherein the core comprises one or more 2'-deoxyribose sugar moieties.

16. 16. The composition of any one of claims 8 to 15, wherein the core comprises one or more 2'-deoxyribose sugar moieties and is capable of annealing to a target nucleic acid, forming a substrate for RNase H, and enabling cleavage of the target nucleic acid by RNase H.

17. 17. The composition of any one of claims 8 to 16, wherein the core comprises five or more 2'-deoxyribose sugar moieties and is capable of annealing to a target nucleic acid, forming a substrate for RNase H, and enabling cleavage of the target nucleic acid by RNase H.

18. 18. The composition of any one of claims 8 to 17, wherein the sugar is 2'-deoxyribose or a bicyclic sugar, and the sugar modification is 2'-MOE, 2'-OMe, or 2'-F.

19. 19. The composition of any one of claims 8 to 18, wherein the internucleotide bond is a phosphodiester bond, a phosphorothioate bond, or a non-negatively charged internucleotide bond.

20. 20. The composition of any one of claims 8 to 19, wherein the oligonucleotide comprises stereorandom internucleotide linkages.

21. 21. The composition of any one of claims 8 to 20, wherein the oligonucleotide comprises a chiral controlled internucleotide bond.

22. 22. The composition of any one of claims 8 to 21, wherein the pattern of backbone chiral centers of the oligonucleotide comprises chiral internucleotide linkages in the Rp configuration and / or chiral internucleotide linkages in the Sp configuration.

23. 23. The composition of any one of claims 8 to 22, wherein the oligonucleotide comprises a chiral internucleotide linkage in the Sp configuration.

24. 24. The composition of any one of claims 8 to 23, wherein the oligonucleotide comprises phosphorothioate linkages in the Sp configuration.

25. 25. The composition of any one of claims 8 to 24, wherein the first wing and the second wing comprise different sugars or sugar modifications or combinations or patterns thereof.

26. 26. The composition of any one of claims 8 to 25, wherein the first wing and the second wing comprise different internucleotide linkages or combinations or patterns thereof.

27. 27. The composition of any one of claims 8 to 26, wherein the first wing and the second wing comprise different stereochemistries or combinations or patterns of internucleotide linkages.

28. 28. The composition of any one of claims 8 to 27, wherein the first wing comprises a sugar or sugar modification that is not present in the core, and the second wing comprises a sugar or sugar modification that is not present in the first wing or the core.

29. 29. The composition of any one of claims 8 to 28, wherein the first wing comprises a first sugar or sugar modification that is not present in the core, the second wing comprises a first sugar or sugar modification that is not present in the first wing or core, and the second wing further comprises a second sugar or sugar modification that is not present in the first wing or core.

30. 30. The composition of any one of claims 8 to 29, wherein the core does not comprise 2'-OMe, the first wing comprises 2'-OMe, and the second wing comprises a 2'-sugar modification that is not 2'-OMe and is not present in the core.

31. 31. The composition of any one of claims 8 to 30, wherein the core does not contain 2'-MOE, the first wing contains 2'-MOE, and the second wing contains a 2'-sugar modification that is not 2'-MOE and is not present in the core.

32. 32. The composition of any one of claims 8 to 31, wherein the core does not contain 2'-F, the first wing contains 2'-F, and the second wing contains a 2'-sugar modification that is not 2'-F and is not present in the core.

33. 33. The composition of any one of claims 8-32, wherein the core does not comprise a bicyclic sugar, the first wing comprises a bicyclic sugar, and the second wing is not a bicyclic sugar and comprises a 2'-sugar modification not present in the core.

34. 34. The composition of any one of claims 8 to 33, wherein the core does not contain 2'-OMe or 2'-MOE, the first wing contains 2'-OMe and 2'-MOE, and the second wing contains 2'-MOE and no 2'-OMe.

35. 35. The composition of any one of claims 8 to 34, wherein the core does not contain 2'-OMe or 2'-MOE, the first wing contains 2'-OMe and 2'-MOE, and the second wing contains 2'-OMe and no 2'-MOE.

36. 36. The composition of any one of claims 8 to 35, wherein the core does not comprise 2'-OMe, 2'-MOE, or 2'-F, the first wing comprises 2'-OMe and 2'-MOE, and the second wing comprises 2'-F and is free of both 2'-MOE and 2'-OMe.

37. 37. The composition of any one of claims 8 to 36, wherein the core does not comprise 2'-OMe, 2'-MOE, or 2'-F, the first wing comprises 2'-OMe and does not comprise 2'-MOE or 2'-F, and the second wing comprises 2'-MOE and does not comprise 2'-OMe or 2'-F.

38. 38. The composition of any one of claims 8 to 37, wherein the core does not comprise 2'-OMe, 2'-MOE, or 2'-F, the first wing comprises 2'-F and does not comprise 2'-MOE or 2'-OMe, and the second wing comprises 2'-OMe and does not comprise 2'-MOE or 2'-F.

39. 39. The composition of any one of claims 8 to 38, wherein the core does not comprise 2'-OMe, 2'-MOE, or 2'-F, the first wing comprises 2'-F and does not comprise 2'-MOE or 2'-OMe, and the second wing comprises 2'-MOE and does not comprise 2'-OMe or 2'-F.

40. a) a common base sequence; b) common backbone bonding patterns; c) Pattern of common skeletal chiral centers In a chiral controlled oligonucleotide composition comprising a plurality of oligonucleotides having the levels of said plurality of oligonucleotides in said composition are non-random; A composition, wherein each oligonucleotide of a particular oligonucleotide type is independently an oligonucleotide or a salt thereof according to any one of claims 1 to 39.

41. a) a common base sequence; b) common backbone bonding patterns; c) Pattern of common skeletal chiral centers In a composition comprising an oligonucleotide of a specific oligonucleotide type characterized by enriched for oligonucleotides of said particular oligonucleotide type compared to a substantially racemic preparation of oligonucleotides having the same common base sequence; A composition, wherein each oligonucleotide of the specific oligonucleotide type is independently an oligonucleotide or a salt thereof according to any one of claims 1 to 39.

42. 43. The composition of claim 41 or 42, wherein at least 0.1%, 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of all oligonucleotides in the composition having a common base sequence are said plurality or type of oligonucleotides.

43. 44. The composition of any one of claims 41 to 43, wherein at least 0.1%, 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of all oligonucleotides in the composition have the common base sequence.

44. 44. The composition of claim 42 or 43, wherein the percentage is at least 5%.

45. 44. The composition of claim 42 or 43, wherein said percentage is at least 10%.

46. 40. A pharmaceutical composition comprising an oligonucleotide according to any one of claims 1 to 39 or a pharmaceutically acceptable salt thereof.

47. 47. The composition of claim 46, comprising a sodium salt of an oligonucleotide of any one of claims 1 to 39.

48. 48. A composition or oligonucleotide according to any one of claims 1 to 47, characterized in that the oligonucleotide of the composition or the oligonucleotide contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more non-negatively charged internucleotide bonds.

49. 1. A method for selectively suppressing transcripts from a target nucleic acid sequence in a population where one or more similar sequences are present, each of the one or more similar sequences containing a signature sequence element of specific nucleotides that defines the target nucleic acid sequence relative to the similar sequence, the method comprising: 1) common base sequence, 2) common backbone bonding patterns; 3) Common skeletal chiral center patterns contacting a chirality-controlled oligonucleotide composition comprising a plurality of oligonucleotides having Including, The method of claim 1, wherein the common base sequence for the oligonucleotides of a particular oligonucleotide type is or includes a sequence complementary to the characteristic sequence element that defines the target nucleic acid sequence.

50. 50. The method of claim 49, wherein the characteristic sequence element is or comprises one or more nucleic acid bases that distinguish the target nucleic acid sequence from similar sequences in the genome and / or in the product encoded thereby.

51. 50. The method of claim 49, wherein a distinctive sequence element is a nucleobase that distinguishes the target nucleic acid sequence from similar sequences in the genome and / or in the product encoded thereby.

52. 52. The method of any one of claims 49-51, wherein a similar sequence shares at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% but less than 100% homology with the target nucleic acid sequence within the portion of the sequence complementary to the common base sequence.

53. 53. The method of any one of claims 49 to 52, wherein the similar sequence differs from, but is not identical to, the target nucleic acid sequence by no more than 5, 4, 3, 2, or 1 nucleic acid base within the portion of the sequence that is complementary to the common base sequence.

54. 53. The method of any one of claims 49 to 52, wherein the analogous sequence differs from the target nucleic acid sequence by only one nucleic acid base within the portion of the sequence that is complementary to the common base sequence.

55. 1. A method for allele-specific suppression of transcripts from a target nucleic acid sequence having multiple alleles present in a population, each of the multiple alleles containing a specific nucleotide signature sequence element that defines the allele relative to other alleles of the same target nucleic acid sequence, the method comprising: 1) common base sequence, 2) common backbone bonding patterns; 3) Common skeletal chiral center patterns contacting a chirality-controlled oligonucleotide composition comprising a plurality of oligonucleotides having Including, The method, wherein the common base sequence for the oligonucleotides of a particular oligonucleotide type is or includes a sequence complementary to the characteristic sequence element that defines a particular allele.

56. 56. The method of any one of claims 49 to 55, wherein the characteristic sequence element is a SNP.

57. 56. The method of any one of claims 49 to 55, wherein the characteristic sequence element is a mutation.

58. 58. The method of any one of claims 49-57, wherein the composition provides a level of repression of the transcript that is greater than in the absence of the composition.

59. 59. The method of any one of claims 49-58, wherein the composition provides repression of the transcript at a level greater than the level of repression observed for another allele or similar sequence.

60. 60. The method of any one of claims 49 to 59, wherein the chiral controlled oligonucleotide composition is a chiral controlled oligonucleotide composition of an oligonucleotide of any one of claims 1 to 39 or 48, or a composition of any one of claims 40 to 48.

61. A method for reducing the level of a transcript or the protein encoded thereby in a system, comprising administering an oligonucleotide or composition described in any one of claims 1 to 48.

62. 432. A compound, oligonucleotide composition or method according to any one of embodiments 1 to 431.

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