5'-modified monomers, oligonucleotides, and double-stranded RNA

Abstract

The technologies described herein relate to 5'-modified nucleosides, nucleotides, oligonucleotides, and double-stranded RNAs, such as siRNA, and kits containing them and methods for using them to inhibit target genes.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims the benefit under §119(e) of U.S. Provisional Application / 521,256 filed June 15, 2023, and incorporates its entire contents in whole by reference herein.

[0002] Technical field The technologies described herein generally relate to 5'-modified nucleosides, nucleotides, oligonucleotides, and double-stranded RNA, such as siRNA, compositions and kits containing them, and methods of using them to inhibit target genes. [Background technology]

[0003] background There is still a demand in the field for oligonucleotides and siRNAs with improved activity and / or pharmacodynamics. This invention addresses some of these demands. [Overview of the project]

[0004] In one aspect, the following is provided: Formula I [ka] [During the ceremony, B is a modified nucleic acid base as desired (e.g., uracil or 5-methyluracil); X is either O or S; Each R V These are independently hydrogen or a hydroxyl protecting group (e.g., ethyl or pivaloyloxymethyl ((CH3)3CC(O)OCH2-,POM); R 2 and R 3 One of them is hydrogen, halogen, or -OR 20 And here: R 20 C is a hydrogen atom, a hydroxyl protecting group, or optionally substituted C 1-6Alkyl (e.g., methyl, 2 - methoxyethyl, 1,3 - dimethoxyprop - 2 - yl, 2-(N - methylamino)-2 - oxoethyl), optionally substituted C 2-6 alkenyl, or optionally substituted C 2-6 alkynyl (e.g., propargyl); R 2 and R 3 the other is -OR 30 where: R 30 is hydrogen, a hydroxy - protecting group, a reactive phosphorus group (e.g., phosphoramidite), a bond to a nucleoside or nucleotide, or a bond to an oligonucleotide. ]] is a compound of or a salt thereof.

[0005] In certain embodiments, X is O. In certain other embodiments, X is S.

[0006] In the compounds of formula I, each R V is independently hydrogen or a hydroxyl - protecting group. In certain embodiments, each R V is hydrogen. In certain embodiments, each R V is a hydroxyl - protecting group such as pivaloyloxymethyl ((CH3)3CC(O)OCH2 -, POM) or ethyl (CH3CH2 -).

[0007] In some of the compounds described herein, R 3 is -OR 30 For example, R 3 is -OR 30 and R 30 is a reactive phosphorus group. Exemplary reactive phosphorus groups are described below and include, but are not limited to, phosphoramidite, H - phosphonate, alkyl - phosphonate, triester phosphate, and phosphorus - containing chiral auxiliaries. Thus, in certain embodiments, R 3 is -OR 30 and R 30 is a reactive phosphorus group selected from phosphoramidite, H - phosphonate, alkyl - phosphonate, and triester phosphate, and optionally R30 It is a phosphoramidite. Therefore, in some compounds, R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P3 C1-C is replaced as desired. 30 Alkyl, optionally substituted C2-C 30 Alkenyl, or optionally substituted C2-C 30 Alkinyl (for example, C1-C which is substituted as desired) 10 Alkyl, optionally substituted C2-C 10 Alkenyl, or optionally substituted C2-C 10 Alkinyl) is; Each R P1 C is independently replaced as desired. 1-6 It is alkyl; and Each R P2 C is independently replaced as desired. 1-6 Alkyl (e.g., methyl, ethyl, propyl, or isopropyl, e.g., isopropyl) or both R P2 These, together with the nitrogen atom to which they are bonded, form a 3- to 8-membered heterocycline which is optionally substituted; or R P1 and R P2 One of them combines to form a 4- to 8-membered heterocyclyl which is combined with the atoms they are attached to and optionally substituted as desired.

[0008] In certain embodiments, R 3 is -OR 30 and R 30 is -P(OR P1 )N(R P2 )2. For example, R 3 is -OR 30 and R 30 is -P(OR[[ID=​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​is C substituted with cyano or -SC(O)Ph 1-6 alkyl, and each R P2 is independently methyl, ethyl, propyl, or isopropyl. For example, R 3 is -OR 30 , and R 30 is -P(OR P1 )N(R P2 )2, where R P1 is -CH2CH2CN, and each R P2 is isopropyl.

[0011] In certain embodiments, R 3 is -OR 30 , and R 30 is a phosphoramidite group such as 3’-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite or 3’-[(β-thiobenzoylethyl)-(1-pyrrolidinyl)]-thiophosphoramidite).

[0012] In other examples, R 3 is -OR 30 , and R 30 is hydrogen or a hydroxyl protecting group (e.g., a silyl-based hydroxyl protecting group). Certain exemplary hydroxyl protecting groups for R 3 of R 30 include, but are not limited to, t-butyldimethylsilyl (TBDMS), trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethyltexylsilyl, t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), or t-butylmethoxyphenylsilyl (TBMPS). Desirably, the hydroxyl protecting group is TBDMS.

[0013] In yet other examples, R 3 is -OR 30 , and R 30This involves binding to a nucleoside or nucleotide, or to an oligonucleotide. For example, R 30 This is binding to an oligonucleotide. 30 When the binding is to a nucleoside or nucleotide, or to an oligonucleotide, R 3 It can bind to the 5'-hydroxyl of a nucleoside or nucleotide, or to the 5'-hydroxyl of the 5' end of an oligonucleotide. It should be noted that the nucleotide bond between the compound of formula I or a nucleoside and the nucleoside, nucleotide, or oligonucleotide bound to it may be an unmodified nucleotide bond (i.e., a phosphodiester) or a modified nucleotide bond (e.g., a phosphorothioate, MMI, or imidp, preferably a phosphorothioate). Exemplary modified nucleotide bonds are listed below.

[0014] One reason, R 3 is hydrogen or a halogen (e.g., F, Br, Cl, or I). For example, R 3 It is either H or F.

[0015] In some compounds of formula I, R 3 は-OR 20 And here, R 20 C is replaced as desired. 1-6 It is alkyl. For example, R 3 は-OR 20 And here, R 20 is methyl, ethyl, or propyl. In one embodiment, R 3 は-OR 20 And here, R 20 It is methyl.

[0016] In some compounds of formula I, R 3 は-OR 20 And here, R 20 C is replaced as desired. 2-6 It is an alkenyl. For example, R 3は-OR 20 And here, R 20 R is vinyl or allyl. In some compounds of formula I, R 3 は-OR 20 And here, R 20 C is replaced as desired. 2-6 It is alkinyl. For example, R 3 は-OR 20 And here, R 20 These are acetylenyl, propargyl, or 5-hexyne-1-yl.

[0017] In some compounds of formula I, R 3 は-OR 20 And here, R 20 is C 1-6 Alkoxy C 1-6 It is alkyl. For example, R 3 は-OR 20 And here, R 20 It is 2-methoxyethyl.

[0018] Preferably, R 3 は-OR 30 And R 2 is hydrogen, halogen, or -OR 20 For example, R 3 は-OR 30 and R 2 is hydrogen, halogen, or -OR 20 And here, R 30 This involves binding to a reactive phosphorus group, a hydroxyl protecting group, a nucleoside or nucleotide, or an oligonucleotide.

[0019] One reason, R 3 は-OR 30 , R 2 R is hydrogen or a halogen (e.g., F, Br, Cl, or I), where R 30 This is a bond to a reactive phosphorus group, a hydroxyl protecting group, a nucleoside or nucleotide, or an oligonucleotide. For example, R 2 It is either H or F.

[0020] In some compounds of formula I, R 2 は-OR 20 And here, R 20 C is replaced as desired. 1-6 It is alkyl. In one example, R 20 is halogen, -OR 22 , -N(R 22 )2, -SR 22 , -C(O)OR 22 ,-C(O)N(R 22 )C substituted with one, two, or three substituents independently selected from the group consisting of 2 1-6 It is alkyl, and here, R 22 is hydrogen or C 1-3 Alkyl (e.g., 2,2,2-trifluoroethyl, 1,3-dimethoxyprop-2-yl). In other examples, R 20 is halogen, -OR 22 , -N(R 22 )2, -SR 22 , -C(O)OR 22 ,-C(O)N(R 22 )C substituted with one or two substituents independently selected from the group consisting of 2 1-6 It is alkyl, and here, R 22 is hydrogen or C 1-3 These are alkyl groups (for example, 2,2,2-trifluoroethyl, 1,3-dimethoxyprop-2-yl).

[0021] In other examples, R 2 は-OR 20 And here, R 20 is methyl, ethyl, or propyl. In one embodiment, R 2 は-OR 20 And here, R 20 It is methyl.

[0022] In some compounds of formula I, R 2 は-OR 20 And here, R 20 is C 1-6 Alkoxy C 1-6It is alkyl. For example, R 2 は-OR 20 And here, R 20 It is 2-methoxyethyl.

[0023] In some compounds of formula I, R 2 は-OR 20 And here, R 20 is N-(C 1-6 Alkyl)aminocarbonyl C 1-6 It is alkyl. For example, R 2 は-OR 20 And here, R 20 It is 2-(N-methylamino)-2-oxoethyl.

[0024] In other examples, R 2 は-OR 30 And R 30 R is a hydrogen or hydroxyl protecting group (e.g., a silyl-based hydroxyl protecting group). 2 R 30 Exemplary hydroxyl protecting groups include, but are not limited to, t-butyldimethylsilyl (TBDMS), trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethyltexylsilyl, t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), or t-butylmethoxyphenylsilyl (TBMPS), and optionally the hydroxyl protecting group is TBDMS.

[0025] In some of the compounds described here, R 2 は-OR 30 For example, R 2 は-OR 30 And R 30R is a reactive phosphorus group. Exemplary reactive phosphorus groups are listed below and include, but are not limited to, phosphoramidites, H-phosphonates, alkyl-phosphonates, phosphate triesters, and phosphorus-containing chiral additives. Therefore, in one embodiment, R 2 は-OR 30 And R 30 R is a reactive phosphorus group selected from phosphoramidites, H-phosphonates, alkyl-phosphonates, and phosphate triesters, and optionally R 30 It is a phosphoramidite. Therefore, in some compounds, R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P3 C1-C is replaced as desired. 30 Alkyl, optionally substituted C2-C 30 Alkenyl, or optionally substituted C2-C 30 Alkinyl (for example, C1-C which is substituted as desired) 10 Alkyl, optionally substituted C2-C 10 Alkenyl, or optionally substituted C2-C 10 Alkinyl) is; Each R P1C is independently replaced as desired. 1-6 It is alkyl; and Each R P2 C is independently replaced as desired. 1-6 Alkyl (e.g., methyl, ethyl, propyl, or isopropyl, e.g., isopropyl) or both R P2 These, together with the nitrogen atom to which they are bonded, form a 3- to 8-membered heterocycline which is optionally substituted; or R P1 and R P2 One of them unites with the atom to which they are bonded, forming a 4- to 8-membered heterocycline which is optionally substituted.

[0026] One reason, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2. For example, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, and here, R P1 C is substituted with cyano or -SC(O)Ph. 1-6 It is alkyl. In one example, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, and here, R P1 It is -CH2CH2CN.

[0027] One reason, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, and each R P2 R is independently methyl, ethyl, propyl, or isopropyl. For example, R 2 は-OR 30 And R30 is -P(OR P1 )N(R P2 )2, where each R P2 It is isopropyl.

[0028] One preferred reason is, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, and here, R P1 C is substituted with cyano or -SC(O)Ph. 1-6 It is alkyl, and each R P2 R is independently methyl, ethyl, propyl, or isopropyl. For example, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, and here, R P1 is -CH2CH2CN, and each R P2 It is isopropyl.

[0029] One reason, R 2 は-OR 30 And R 30 This refers to a phosphoramidite group such as 2'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite or 2'-[(β-thiobenzoylethyl)-(1-pyrrolidinyl)]-thiophosphoramidite).

[0030] In yet another example, R 2 は-OR 30 And R 30 This involves binding to a nucleoside or nucleotide, or to an oligonucleotide. For example, R 30 This is binding to an oligonucleotide. 30 When the binding is to a nucleoside or nucleotide, or to an oligonucleotide, R 2It can bind to the 5'-hydroxyl of a nucleoside or nucleotide, or to the 5'-hydroxyl of the 5' end of an oligonucleotide. It should be noted that the nucleotide bond between the compound of formula I or a nucleoside and the nucleoside, nucleotide, or oligonucleotide bound to it may be an unmodified nucleotide bond (i.e., a phosphodiester) or a modified nucleotide bond (e.g., a phosphorothioate, MMI, or imidp, preferably a phosphorothioate). Exemplary modified nucleotide bonds are listed below.

[0031] In the compounds of formula I, B is a naturally occurring or unnatural nucleic acid base that is optionally modified. For example, B is uracil, adenine, cytosine, 5-methylcytosine, guanine, or thymine (i.e., 5-methyluracil). In some embodiments, B is a modified or protected nucleic acid base. For example, B is a protected nucleic acid base containing at least one amine or hydroxyl protecting group. In some embodiments, B is adenine, cytosine, 5-methylcytosine, or guanine containing at least one amine protecting group. Exemplary modified, unmodified, and unnatural nucleic acid bases are listed below.

[0032] In some compounds of formula I, X is O; R 2 is hydrogen, F, or -OR 20 And here, R 20 C is substituted with hydrogen, if desired. 1-6 Alkyl (e.g., methyl, ethyl, or propyl), e.g., C 1-6 Alkoxy C 1-6 Alkyl (e.g., methoxy, 2-methoxyethyl) or N-(C) 1-6 Alkyl)aminocarbonyl C 1-6 It is alkyl (e.g., 2-(N-methylamino)-2-oxoethyl); and R 3 は-OR 30 And here, R 30 This is hydrogen, a hydroxyl protecting group, or a reactive phosphorus group.

[0033] In some compounds of formula I, X is O; R 2 is hydrogen, F, or -OR 20 And here, R 20 C is substituted with hydrogen, if desired. 1-6 Alkyl (e.g., methyl, ethyl, or propyl), e.g., C 1-6 Alkoxy C 1-6 Alkyl (e.g., methoxy, 2-methoxyethyl) or N-(C) 1-6 Alkyl)aminocarbonyl C 1-6 It is alkyl (for example, 2-(N-methylamino)-2-oxoethyl); and R 3 は-OR 30 And here, R 30 is a reactive phosphorus group (e.g., phosphoramidite, H-phosphonate, alkyl-phosphonate, or phosphoric acid triester). In one further embodiment, the reactive phosphorus group is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 (For example, -P(OR P1 )N(R P2 )2) and here: Each R P3 C1-C is replaced as desired. 30 Alkyl, optionally substituted C2-C 30Alkenyl, or optionally substituted C2-C 30 Alkinyl (for example, C1-C which is substituted as desired) 10 Alkyl, optionally substituted C2-C 10 Alkenyl, or optionally substituted C2-C 10 Alkinyl) and each R P1 C is independently replaced as desired. 1-6 Alkyl (e.g., C substituted with cyano or -SC(O)Ph) 1-6 Alkyl (e.g., 2-cyanoethyl); and each R P2 C is independently replaced as desired. 1-6 Alkyl (e.g., methyl, ethyl, propyl, or isopropyl, preferably isopropyl), or both R P2 They form a 3- to 8-membered heterocycline which is optionally substituted with the nitrogen atom to which they are bonded; or R P1 and R P2 One of them unites with the atom to which they are bonded, forming a 4- to 8-membered heterocycline which is optionally substituted.

[0034] In some compounds of formula I, X is O; R 2 is hydrogen, F, or -OR 20 And here, R 20 C is substituted with hydrogen, if desired. 1-6 Alkyl (e.g., methyl, ethyl, or propyl, preferably methyl), for example, C 1-6 Alkoxy C 1-6 Alkyl (e.g., methoxy, 2-methoxyethyl) or N-(C) 1-6 Alkyl)aminocarbonyl C 1-6 It is alkyl (for example, 2-(N-methylamino)-2-oxoethyl); and R 3 は-OR 30 And here, R 30 is -P(OR P1 )N(R P2 )2, and here, R P1 is -CH2CH2CN, and each R P2It is isopropyl.

[0035] In one preferred embodiment, X is O; R 2 is hydrogen, F, or -OR 20 And here, R 20 R is hydrogen, methyl, 2-methoxyethyl, 1,3-dimethoxyprop-2-yl, or 2-(N-methylamino)-2-oxoethyl; and R 3 は-OR 30 And here, R 30 is -P(OR P1 )N(R P2 )2, and here, R P1 is -CH2CH2CN, and each R P2 It is isopropyl.

[0036] In one embodiment, the compound is of formula [ka] [During the ceremony, B is a modified nucleic acid base (e.g., uracil) as desired; X is either O or S; Each R V These are independently hydrogen or a hydroxyl protecting group (e.g., ethyl or pivaloyloxymethyl); R 2 and R 3 One of them is hydrogen, halogen, or -OR 20 And here: R 20 C is a hydrogen atom, a hydroxyl protecting group, or optionally substituted C 1-6 Alkyl (e.g., methyl, 2-methoxyethyl, 1,3-dimethoxyprop-2-yl, 2-(N-methylamino)-2-oxoethyl), optionally substituted C 2-6 alkenyl, or C substituted as desired. 2-6 Alkinyl (for example, propargyl); and R 2 and R 3 The other side is -OR 30 And here: R 30 This involves binding to hydrogen, a hydroxyl protecting group, a reactive phosphorus group (e.g., a phosphoramidite), a nucleoside or nucleotide, or an oligonucleotide. It is that or its salt.

[0037] In one embodiment, the compound is of formula [ka] [During the ceremony, B is a modified nucleic acid base (e.g., uracil) as desired; X is either O or S; Each R V These are independently hydrogen or a hydroxyl protecting group (e.g., ethyl or pivaloyloxymethyl); R 2 and R 3 One of them is hydrogen, halogen, or -OR 20 And here: R 20 C is a hydrogen atom, a hydroxyl protecting group, or optionally substituted C 1-6 Alkyl (e.g., methyl, 2-methoxyethyl, 1,3-dimethoxyprop-2-yl, 2-(N-methylamino)-2-oxoethyl), optionally substituted C 2-6 alkenyl, or C substituted as desired. 2-6 Alkinyl (for example, propargyl); and R 2 and R 3 The other side is -OR 30 And here: R 30 This involves binding to hydrogen, a hydroxyl protecting group, a reactive phosphorus group (e.g., a phosphoramidite), a nucleoside or nucleotide, or an oligonucleotide. It is that or its salt.

[0038] In one embodiment, the compound is formula [ka] It is that or its salt.

[0039] In one embodiment, the compound is formula [ka] It is that or its salt.

[0040] In one embodiment, the compound is formula [ka] It is that or its salt.

[0041] One reason, R 3 は-OR 30 For example, a compound is given by formula [ka] [In the formula: R 3 は-OR 30 And here: R 30 This can be a bond to hydrogen, a hydroxyl protecting group, a reactive phosphorus group (e.g., a phosphoramidite), a nucleoside or nucleotide, or an oligonucleotide; B is a modified nucleic acid base (e.g., uracil) as desired; X is either O or S; Each R P is independently a hydrogen or hydroxyl protecting group (e.g., ethyl or pivaloyloxymethyl); and R 2 is hydrogen, halogen, or -OR 20 And here: R 20 C is a hydrogen atom, a hydroxyl protecting group, or optionally substituted C 1-6 Alkyl (e.g., methyl, 2-methoxyethyl, 1,3-dimethoxyprop-2-yl, 2-(N-methylamino)-2-oxoethyl), optionally substituted C 2-6alkenyl, or C substituted as desired. 2-6 This refers to alkinyl (for example, propargyl). It is that or its salt.

[0042] In one embodiment, B is uracil or thymine. In one embodiment, R V R is hydrogen. In one way, V The protecting group is a hydroxyl protecting group (e.g., ethyl or pivaloyloxymethyl).

[0043] One reason, R 3 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, and X is O. For example, compound formula [ka] [In the formula: R 3’ は-OR 30 And here: R 30 is -P(OR P1 )N(R P2 )2, Here: Each R P1 C is replaced as desired. 1-6 It is an alkyl group (e.g., -CH2CH2CN); Each R P2 C is independently replaced as desired. 1-6 Alkyl (e.g., isopropyl); X is O; B is a modified nucleic acid base (e.g., uracil) as desired; Each R P is independently a hydrogen or hydroxyl protecting group (e.g., ethyl or pivaloyloxymethyl); and R 2’ is hydrogen, halogen, or -OR 20 And here: R 20C is a hydrogen atom, a hydroxyl protecting group, or optionally substituted C 1-6 Alkyl (e.g., methyl, 2-methoxyethyl, 1,3-dimethoxyprop-2-yl, 2-(N-methylamino)-2-oxoethyl), optionally substituted C 2-6 alkenyl, or C substituted as desired. 2-6 This refers to alkinyl (for example, propargyl). It is that or its salt.

[0044] In one embodiment, B is uracil or thymine. In one embodiment, R V R is hydrogen. In one way, V The protecting group is a hydroxyl protecting group (e.g., ethyl or pivaloyloxymethyl).

[0045] In a compound of formula (I), [ka] part [ka] It is replaced by, that is, the compound is given the formula [ka] These are, and here, B, X, R V , R 2 and R 3 The formula (I) is defined here. For example, the compound has the structure: [ka] It belongs to them.

[0046] In one embodiment, the compound is of formula [ka] [During the ceremony, B is a modified nucleic acid base (e.g., uracil) as desired; X is either O or S; Each R V is independently a hydrogen or hydroxyl protecting group (e.g., ethyl or pivaloyloxymethyl); R 2 is hydrogen, halogen, or -OR 20 And here: R 20 C is a hydrogen atom, a hydroxyl protecting group, or optionally substituted C 1-6 Alkyl (e.g., methyl, 2-methoxyethyl, 1,3-dimethoxyprop-2-yl, 2-(N-methylamino)-2-oxoethyl), optionally substituted C 2-6 alkenyl, or C substituted as desired. 2-6 Alkinyl (for example, propargyl); and [ka] This indicates the binding of the oligonucleotide to the rest of the molecule (e.g., the antisense strand of a double-stranded RNA molecule). It is that or its salt.

[0047] In the foregoing, the oxygen atom described as linking the 5' end of the oligonucleotide to the phosphorus atom is the 5'-oxygen of the 5' terminal nucleoside of the oligonucleotide.

[0048] In one embodiment, Y is S. In another embodiment, Y is O.

[0049] Here, [ka] The term is used to represent oligonucleotides; such oligonucleotides may be RNA, DNA, single-stranded RNA, e.g., antisense oligonucleotides (ASOs), antisense strands of double-stranded RNA (e.g., siRNAs), and oligonucleotide derivatives, e.g., phosphorodiamidate morpholino oligomers (PMOs).

[0050] In one embodiment, the compound is of formula [ka] [During the ceremony, R V This is a hydrogen or hydroxyl protecting group (e.g., ethyl or pivaloyloxymethyl). It is a salt of or the same. In one embodiment, B is uracil or thymine. In one embodiment, R V R is hydrogen. In one way, V is a hydroxyl protecting group (e.g., pivaloyloxymethyl). In one embodiment, Y is S. In another embodiment, Y is O.

[0051] In one embodiment, the compound is selected from the group of compounds shown in Table A: [Table 1] [Table 2]

[0052] In another aspect, the structure provided herein is: [ka] [During the ceremony: X is either O or S; and Each R V [These are independently hydrogen or hydroxyl protecting groups.] It is an oligonucleotide having a 5'-terminus modification that includes [a specific compound].

[0053] In one embodiment, the 5'-terminal modification is structural: [ka] Includes.

[0054] In one embodiment, X is O. In another embodiment, X is O, and each RV is hydrogen. In one embodiment, X is O, and each R V is ethyl. In one embodiment, X is O, and each R V It is pivaloyloxymethyl.

[0055] [ka] It should be noted that the sugar portion of a modified nucleotide, i.e., the nucleotide at the 5' end of an oligonucleotide (5' terminal nucleotide), may contain a 5-membered or 6-membered ring. For example, the sugar portion of the modified nucleotide may be a furanose (e.g., ribofuranose, arabinofuranose, lyxofuranose, xylofuranose, librofuranose, or xylulofuranose, including its alpha and beta, D and L, deoxy, and modified derivatives) or a pyranose (e.g., glucopyranose, galactopyranose, mannopyranose, allopyranose, altropyranose, globyranose, idopyranose, and taropyranose, including its alpha and beta, D and L, deoxy, and modified derivatives).

[0056] in general, [ka] The modification replaces the CH2OH group in the sugar portion of the 5' terminal nucleotide of the oligonucleotide. For example, [ka] The modification replaces the 4'-CH2OH group of the furanose ring (e.g., ribofuranose, arabinofuranose, lyxofuranose, xylofuranose, librofuranose, or xylulofuranose, preferably ribofuranose) at the 5' terminal nucleotide of the oligonucleotide, or the 5'-CH2OH group of the pyranose (e.g., glucopyranose, galactopyranose, mannopyranose, allopyranose, altropyranose, globyranose, idopyranose, or taropyranose, preferably glucopyranose, galactopyranose, or mannopyranose).

[0057] In one embodiment, the oligonucleotide contains a compound of formula (I) described herein at its 5' end. For example, the 5' terminal nucleotide of the oligonucleotide has the structure: [ka] [During the ceremony, Each R V These are independently hydrogen or a hydroxyl protecting group (e.g., ethyl or pivaloyloxymethyl ((CH3)3CC(O)OCH2-,POM); B is a modified nucleic acid base (e.g., uracil) as desired; R 2 and R 3 One of them is hydrogen, halogen, or -OR 20 And here: R 20 C is a hydrogen atom, a hydroxyl protecting group, or optionally substituted C 1-6 Alkyl (e.g., methyl, 2-methoxyethyl, 1,3-dimethoxyprop-2-yl, 2-(N-methylamino)-2-oxoethyl), optionally substituted C 2-6 alkenyl, or C substituted as desired. 2-6 Alkinyl (for example, propargyl); and R 2 and R 3 The other side is -OR 30 And here: R 30This refers to binding to an oligonucleotide (for example, an internucleotide bond that connects to a subsequent nucleotide of an oligonucleotide). It belongs to them.

[0058] In one embodiment, the 5' terminal nucleotide of the oligonucleotide is structured as follows: [ka] It has, and here R 2 and R 3 One of them is -OR 30 (For example, R 3 は-OR 30 ) and here:R 30 This refers to binding to an oligonucleotide (for example, an internucleotide bond that connects to a subsequent nucleotide of the oligonucleotide).

[0059] In another embodiment, the 5' terminal nucleotide of the oligonucleotide has the following structure: [ka] It has, and here R 2 and R 3 One of them is -OR 30 (For example, R 3 は-OR 30 ) and here:R 30 This refers to binding to an oligonucleotide (for example, an internucleotide bond that connects to a subsequent nucleotide of the oligonucleotide).

[0060] In another embodiment, the 5' terminal nucleotide of the oligonucleotide has the following structure: [ka] It has, and here R 2 and R 3 One of them is -OR 30 (For example, R 3 は-OR 30 ) and here:R 30This refers to binding to an oligonucleotide (for example, an internucleotide bond that connects to a subsequent nucleotide of the oligonucleotide).

[0061] In another embodiment, the 5' terminal nucleotide of the oligonucleotide has the following structure: [ka] It has, and here R 2 and R 3 One of them is -OR 30 (For example, R 3 は-OR 30 ) and here:R 30 This refers to binding to an oligonucleotide (for example, an internucleotide bond that connects to a subsequent nucleotide of the oligonucleotide).

[0062] In another embodiment, the 5' terminal nucleotide of the oligonucleotide has the following structure: [ka] It has, and here R 2 and R 3 One of them is -OR 30 (For example, R 3 は-OR 30 ) and here:R 30 This refers to binding to an oligonucleotide (for example, an internucleotide bond that connects to a subsequent nucleotide of the oligonucleotide).

[0063] In general, the oligonucleotides described herein contain at least three nucleotides. For example, an oligonucleotide may contain 5 to 100 nucleotides, e.g., 10 to 50 nucleotides. In one embodiment, an oligonucleotide may contain 15 to 40 nucleotides. For example, an oligonucleotide may be 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides long. In one embodiment, an oligonucleotide may be 17, 18, 19, 21, 22, 23, 24, or 25 nucleotides long. For example, an oligonucleotide may be 19, 20, 21, 22, or 23 nucleotides long. It should be noted that the compound of formula I is counted as one nucleotide.

[0064] The oligonucleotides described herein may contain at least one nucleic acid modification (e.g., one, two, three, four, five, six, seven, eight, nine, ten or more independently selected modifications). Exemplary nucleic acid modifications are listed below and include, but are not limited to, nucleic acid base modifications, sugar modifications, internucleotide bond modifications, conjugates (e.g., ligands), and combinations thereof.

[0065] In one embodiment, the oligonucleotide comprises at least one (e.g., one, two, three, four, five, six, seven, eight, nine, ten or more) 2'-OMe nucleotide.

[0066] In one embodiment, the oligonucleotide comprises at least one (e.g., one, two, three, four, five, six, seven, eight, nine, ten or more) double-stranded thermal destabilization modification. For example, the oligonucleotide comprises a thermal destabilization modification at at least one position of 4, 5, 6, 7, or 8, where the compound of formula I is at position 1 of the 5' end of the oligonucleotide; optionally, the thermal destabilization modification is located at position 6, 7, or 8, counting from the 5' end of the oligonucleotide, preferably at position 7.

[0067] In one embodiment, the oligonucleotide contains at least one (e.g., one, two, three, four, five, six, seven, eight, nine, ten or more) 2'-F nucleotides. For example, the oligonucleotide contains two, three, four, five, or six 2'-F nucleotides, and optionally, the oligonucleotide contains three, four, five, or six 2'-F nucleotides. In one embodiment, the oligonucleotide contains at least 2'-F nucleotides at positions 2, 14, and 16, where counted from the 5' end of the oligonucleotide, the compound of formula I is at position 1 of the 5' end of the oligonucleotide. For example, an oligonucleotide may contain 2'-F nucleotides at at least positions 2, 6, 14, and 16, counting from the 5' end of the oligonucleotide; preferably, an oligonucleotide may contain 2'-F nucleotides at at least positions 2, 6, 9, 14, and 16, counting from the 5' end of the oligonucleotide. It should be noted that when an oligonucleotide contains more than one 2'-F nucleotide, each 2'-F nucleotide is an independently selected nucleotide.

[0068] Oligonucleotides may also contain one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2'-deoxy(2'-H) nucleotides. For example, an oligonucleotide may contain 2, 3, 4, 5, 6, or 7 2'-deoxynucleotides, and optionally, an oligonucleotide may contain 3, 4, 5, or 6 2'-deoxynucleotides. An oligonucleotide may contain a 2'-deoxynucleotide at any one of the positions 2, 5, 7, 12, 14, and 16, where counted from the 5' end of the oligonucleotide, the compound of formula I is at position 1 of the 5' end of the oligonucleotide. For example, an oligonucleotide may contain a 2'-deoxynucleotide at at least position 5, counted from the 5' end of the oligonucleotide. In one embodiment, an oligonucleotide may contain a 2'-deoxynucleotide at at least positions 2, 5, and 9, counted from the 5' end of the oligonucleotide. For example, an oligonucleotide contains 2'-deoxynucleotides at at least positions 2, 5, 7, and 12, counting from the 5' end of the oligonucleotide. It should be noted that when an oligonucleotide contains more than one 2'-deoxynucleotide, each 2'-deoxynucleotide is an independently selected nucleotide.

[0069] One or more nucleic acid bases (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more) in an oligonucleotide may be unnatural or modified nucleic acid bases. For example, an oligonucleotide may contain one or more modified or protected nucleic acid bases (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more).

[0070] The nucleotide internucleotide bonds of an oligonucleotide can be independently unmodified (e.g., phosphodiester) or modified (e.g., phosphorothioate). Therefore, in some embodiments, an oligonucleotide contains at least one (e.g., one, two, three, four, five, six, seven, eight, nine, ten or more) modified nucleoside bonds. Generally, an oligonucleotide contains at least one (e.g., one, two, four, or five) modified nucleoside bonds (e.g., phosphorothioate) at the first 1-5 positions of one or both oligonucleotides. For example, an oligonucleotide contains a modified oligonucleotide bond (e.g., phosphorothioate) between nucleotides 1 and 2, and between nucleotides 2 and 3, counting from the 5' end of the oligonucleotide; and an oligonucleotide contains a modified oligonucleotide bond (e.g., phosphorothioate) between nucleotides 1 and 2, and between nucleotides 2 and 3, counting from the 3' end of the oligonucleotide.

[0071] In one embodiment, the oligonucleotide is covalently bonded to a support, such as a solid support.

[0072] In yet another embodiment, provided herein is a double-stranded RNA (dsRNA) comprising a sense strand and an antisense strand, wherein the sense strand is substantially complementary to the antisense strand, and where either the sense strand or the antisense strand is an oligonucleotide described herein, i.e., structure: [ka] [During the ceremony: X is either O or S; and Each R V [These are independently hydrogen or hydroxyl protecting groups.] It is an oligonucleotide having a 5'-terminus modification that includes [a specific compound].

[0073] Preferably, the antisense chain includes the above 5' terminal modification.

[0074] In yet another embodiment, a method for reducing the expression of a target gene in a subject is provided herein. The method comprises administering to the subject (i) a double-stranded RNA described herein (where the antisense strand is substantially complementary to the target gene); or (ii) an oligonucleotide described herein (where the oligonucleotide is substantially complementary to the target gene).

[0075] In other embodiments, provided herein are pharmaceutical compositions comprising the oligonucleotides or dsRNA molecules described herein, either alone or in combination with pharmaceutically acceptable carriers or additives.

[0076] In yet another embodiment, provided herein are cells containing the oligonucleotide or dsRNA molecules described herein.

[0077] In yet another embodiment, provided herein is a gene silencing kit comprising the oligonucleotide or dsRNA molecule described herein.

[0078] Also provided herein is a method for silencing a target gene in cells. The method comprises the steps of (i) introducing a dsRNA molecule as described herein (wherein one strand of the dsRNA, e.g., the antisense, contains a nucleotide sequence substantially complementary to the nucleotide sequence of the target gene); and / or (ii) introducing an oligonucleotide as described herein (wherein the oligonucleotide contains a nucleotide sequence substantially complementary to the nucleotide sequence of the target gene) into cells.

[0079] In another embodiment, provided herein are methods for inhibiting or reducing the expression of a target gene in a subject. The method comprises administering to a subject (i) a dsRNA molecule described herein (wherein one of the strands of the dsRNA, e.g., the antisense, contains a nucleotide sequence substantially complementary to the nucleotide sequence of the target gene); and / or (ii) an oligonucleotide described herein (wherein the oligonucleotide contains a nucleotide sequence substantially complementary to the nucleotide sequence of the target gene). [Brief explanation of the drawing]

[0080] [Figure 1A-1F] A synthesis scheme for the synthesis of some exemplary compounds of the present invention is described below.

[0081] [Figure 2] The present invention targets mTTR and exhibits RNAi activity of some exemplary dsRNAs, including the exemplary compounds of the present invention.

[0082] [Figure 3] This shows the SOD1 evaluation of exemplary dsRNAs, including the example, in rats.

[0083] [Figure 4-7]Molecular modeling tests are shown. Figure 4 shows that 6'-E-VP-RNA occupies a dense space, occupying the position of the water molecule in the parent structure. Figure 5 shows that the spacing is extremely narrow, and the water inserted there in the 4F3T(PDB) miR-20a complex with Ago2 is removed. Otherwise, there would be no room at all between the phosphate inserted slightly deeper into the pocket and the various basic side chains. The reference structure is superimposed on all the others and is colored gray. Figure 6 shows that the 6'-Z-VP modification is well accommodated; it appears to be better accommodated than the 6'-E-VP isomer, which has structural collisions with Gln-545 and Tyr-529; and 6'-Z-VP maintains better stacking interaction with Tyr-529 than 5'-Z-VP. Figure 7 shows that the 6'-Z-VP-U modification of AS1 fits well into the MID binding pocket—it appears better than the 6'-E-VP example, which pushes the terminal phosphate a little too close to multiple basic residues as well as Gln-545 and Tyr-529. [Modes for carrying out the invention]

[0084] It should be understood that both the general statements above and the detailed statements below are merely illustrative and descriptive and do not limit the claimed invention. Here, the singular ice field includes plural ice fields unless otherwise specified. The use of “or” here includes “and / or” unless otherwise specified. Furthermore, the use of the term “including” and other forms such as “containing” and “encompassing” is not limiting. Also, terms such as “element” or “component” include both elements and components containing one unit and elements and components containing more than one subunit unless otherwise specified.

[0085] The section titles used herein are for organizational purposes only and should not be interpreted as limitations on the means described. Patents, patent applications, articles, books, and professional texts. All documents or parts of documents cited in this application, including, are incorporated herein by reference in their entirety for any purpose.

[0086] R 3 In the various embodiments described herein, R 3 is hydrogen, halogen, -OR 20 , or -OR 30 It is possible.

[0087] One reason, R 3 は-OR 30 And here, R 30 This can be a bond to hydrogen, a hydroxyl protecting group, a reactive phosphorus group (e.g., a phosphoramidite), a nucleoside or nucleotide, or an oligonucleotide. For example, R 3 は-OR 30 And R 30 is a hydrogen or hydroxyl protecting group.

[0088] One reason, R 3 は-OR 30 And R 30 R is a reactive phosphorus group. For example, R 30 is a phosphoramidite, H-phosphonate, alkyl-phosphonate, or phosphate triester. In one embodiment, R 3 は-OR 30 and R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SRP1 )R P3 And here: Each R P1 These are independently OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, NH(C1-C4)alkyl, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8)alkyl, O(C1-C8)alkyl (i.e., C1-C8 Alkoxy), O(C1-C8)haloalkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) p C is substituted with one, two, three, four, or five substituents independently selected from -NH2 or CH2-aryl-alkoxy. 1-6 It is an alkyl group, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6; Each R P2Depending on the desired type, these are OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, NH(C1-C4)alkyl, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8)alkyl, O(C1-C8)alkyl (i.e., C1-C 8 alkoxy), O(C1-C8) haloalkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) p C is substituted with one, two, three, four, or five substituents independently selected from -NH2 or CH2-aryl-alkoxy. 1-6 It is an alkyl group, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6, and each R is optional. P2 R is independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, preferably each R P2 It is isopropyl; or both R P2 These, together with the nitrogen atom to which they are bonded, form a 3- to 8-membered heterocycline which is optionally substituted; or R P1 and R P2 One of them unites with the atom to which they are bonded, forming a 4- to 8-membered heterocycline which is optionally substituted; and Each R P3 C is independent 1-30 Alkyl, C2-C 30 Alkenyl, or C2-C 30These are alkynyl compounds, each independently of choice being OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, NH(C1-C4)alkyl, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8)alkyl, O(C1-C8)alkyl(suna (C1-C8 alkoxy), O(C1-C8) haloalkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) p Substituted with one, two, three, four, or five substituents selected from -NH2 or CH2-aryl-alkoxy, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6, and each R optionally P3 The elements are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be optionally substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy.

[0089] One reason, R 3 は-OR 30 and R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1)N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 C is replaced with CN or -SC(O)Ph as desired. 1-6 It is alkyl; each R P2 C is independently replaced as desired. 1-6 It is alkyl; and each R P3 C is independently replaced as desired. 1-6 It is alkyl.

[0090] One reason, R 3 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 C is replaced with CN or -SC(O)Ph as desired. 1-6 It is alkyl; each R P2R is independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl; and each R P3 The elements are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be optionally substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy.

[0091] One reason, R 3 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 It is 2-cyanoethyl (-CH2CH2CN); each R P2 C is independently replaced as desired. 1-6 It is alkyl; and each R P3 C is independently replaced as desired. 1-6 It is alkyl.

[0092] One reason, R 3 は-OR 30 And R 30 is -P(OR P1 )N(R P2)2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 It is 2-cyanoethyl (-CH2CH2CN); each R P2 R is independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl; and each R P3 The elements are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be optionally substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy.

[0093] One reason, R 3 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1)H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 It is 2-cyanoethyl (-CH2CH2CN); each R P2 is independently isopropyl; and each R P3 The elements are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be optionally substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy.

[0094] One reason, R 3 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, where:R P1 2-cyanoethyl (-CH2CH2CN); each R P2 It is independently isopropyl.

[0095] One reason, R 3 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(ORP1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 C is replaced with CN or -SC(O)Ph as desired. 1-6 Alkyl; both R P2 They form 3- to 8-membered heterocyclines which are optionally substituted with the nitrogen atom to which they are bonded; and each R P3 C is independently replaced as desired. 1-6 It is alkyl.

[0096] One reason, R 3 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 C is replaced with CN or -SC(O)Ph as desired. 1-6 Alkyl; both R P2 They form 3- to 8-membered heterocyclines which are optionally substituted with the nitrogen atom to which they are bonded; and each R P3The elements are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be optionally substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy.

[0097] One reason, R 3 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 It is 2-cyanoethyl (-CH2CH2CN); both R P2 They form 3- to 8-membered heterocyclines which are optionally substituted with the nitrogen atom to which they are bonded; and each R P3 C is independently replaced as desired. 1-6 It is alkyl.

[0098] One reason, R 3 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1)N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 It is 2-cyanoethyl (-CH2CH2CN); both R P2 They form 3- to 8-membered heterocyclines which are optionally substituted with the nitrogen atom to which they are bonded; and each R P3 The elements are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be optionally substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy.

[0099] One reason, R 3 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )RP3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 , where: R P1 and R P2 combine to form a 4- to 8-membered heterocyclyl optionally substituted together with the atom to which they are attached; the other of R P2 is independently optionally substituted C 1-6 alkyl; and each R P3 is independently optionally substituted C 1-6 alkyl.

[0100] In certain embodiments, R 3 is -OR 30 , R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR s P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 , where: R P1 s and R P2 combine to form a 4- to 8-membered heterocyclyl optionally substituted together with the atom to which they are attached; the other of R P2 is independently methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, pentyl or hexyl; and each RP3 C is independently replaced as desired 1-6 is alkyl

[0101] In certain embodiments, R 3 is -OR 30 and R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 where: one of R P1 and R P2 combines to form a 4- to 8-member heterocyclyl that is optionally substituted and is combined with the atom to which they are attached; the other of R P2 is independently optionally substituted C 1-6 alkyl; and each R P3 is independently methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, pentyl or hexyl, each of which may optionally be substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy

[0102] In certain embodiments, R 3 is -OR 30 and R 30 is -P(OR P1 )N(R P2 )2, -P(SRP1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here:R P1 and R P2 One of them unites with the atom to which they are bonded, forming a 4- to 8-membered heterocycline which is optionally substituted; R P2 The other is independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl; and each R P3 The elements are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be optionally substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy.

[0103] One reason, R 3 は-OR 30 And R 30 R is a binding to a nucleoside or nucleotide, or an oligonucleotide. 30 When the bond is to a nucleoside, nucleotide, or oligonucleotide, the internucleotide bond between the compound of formula I and the nucleoside, nucleotide, or oligonucleotide can be an unmodified (e.g., phosphodiester) internucleotide bond or a modified (e.g., phosphorothioate) internucleotide bond.

[0104] In one embodiment, R 3 is -OR 30 and R 30 is a nucleoside, nucleotide, or oligonucleotide linked by an unmodified (e.g., phosphodiester) internucleotide linkage (e.g., or a modified (e.g., phosphorothioate) internucleotide linkage at the 5'-position. For example, R 30 is linked to the 5'-end (e.g., 5'-OH) of an oligonucleotide. In one embodiment, R 30 is linked to the 5'-end (e.g., 5'-OH) of an oligonucleotide by an unmodified (e.g., phosphodiester) internucleotide linkage. In some other embodiments, R 30 is linked to the 5'-end (e.g., 5'-OH) of an oligonucleotide by a modified (e.g., phosphorothioate) internucleotide linkage.

[0105] In one embodiment, R 3’ is -OR 30 and R 30 is a hydroxyl protecting group. For example, R 3’ is -OR 30 and R 30This is BOC or Boc, MOM, MTM, t-butylthiomethyl, SMOM, BOM, PMBM, p-AOM, GUM, t-butoxymethyl, POM, siloxymethyl, MEM, 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, SEMOR, THP, 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, MTHP, 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl-S,S-dioxide, CTMP, 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-meth Nobenzofuran-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-trichloroethyl, 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'-dinitrobenzhydryl, 5-dibenzosberyl, 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(ruberinoyloxyphenyl)methyl, 4,4',4”-tris(benzoyloxyphenyl)methyl, 3-(imidazole-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-Benzodisulfuran-2-yl, benzisothiazolyl-S,S-dioxide, TMS, TES, TIPS, IPDMS, DEIPS, dimethyltexylsilyl, TBDMS, TBDPS, tripenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, DPMS, TBMPS, formate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate Triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (rebrinate), 4,4-(ethylenedithio)pentanoate (ruberinoyl dithioacetal), adamantoate, crotonate, 4-methoxycrotonate, bezoate, p-phenylbezoate, 2,4,6-trimethylbezoate (mesitoate), alkyl Methyl carbonate, Fmoc, alkyl ethyl carbonate, Troc, TMSEC, Psec, Peoc, alkyl isobutyl carbonate, alkyl vinyl carbonate, alkyl allyl 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-benzyl thiocarbonate, 4-ethoxy-1-naphthyl carbonate, methyl Dithiocarbonate, 2-iodobezoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)bezoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)bezoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, (E)-2-methyl-2-butanoate, o-(methoxyacyl)bezoate, α-naphthoate, nitrate, alkyl N,N,N',The hydroxyl protecting group is selected from the group consisting of N'-tetramethylphosphodiamide, alkyl N-phenylcarbamate, borate, dimethylphosphinothionel, alkyl 2,4-dinitrophenyl sulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate.

[0106] One reason, R 3’ は-OR 30 And R 30 The compounds are TBDMS, TBDPS, TMS, TES, TIPS, IPDMS, DEIPS, TBMPS, DPMS, dimethyltexylsilyl, tribenzylsilyl, tri-p-xylylsilyl, or triphenylsilyl, and optionally R 30 This is TBDMS.

[0107] One reason, R 3 is hydrogen or halogen. For example, R 3 It is either H or F.

[0108] One reason, R 3 は-OR 20 And here, R 20 C is a hydrogen atom, a hydroxyl protecting group, or optionally substituted C 1-6 Alkyl, for example, C 1-6 Alkoxy C 1-6 Alkyl (e.g., 2-methoxyethyl) or N-(C) 1-6 Alkyl)aminocarbonyl C 1-6 It is an alkyl group (for example, 2-(N-methylamino)-2-oxoethyl). For example, R 3 は-OR 20 And R 20 is a hydrogen or hydroxyl protecting group.

[0109] One reason, R 3Depending on your preference, these can be OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-4)alkyl, SO2NH(C1-4)alkyl, halogen, carbonyl, thiol, cyano, NH2, NH(C1-C4)alkyl, N[(C1-4)alkyl]2, C(O)NH2, C(O)NH(C 1-6 Alkyl), C(O)N(C 1-6 Alkyl)2, COOH, COO(C 1-6 Alkyl) (e.g., COOMe), C 2-6 Acyl (e.g., acetyl), (C1-8)alkyl, O(C1-8)alkyl (i.e., C1-8 alkoxy), O(C1-8)haloalkyl, (C2-8)alkenyl, (C2-8)alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) p - substituted with one, two, three, four, or five substituents independently selected from -NH2 or CH2-aryl-alkoxy -OR 20 And R 20 is C 1-6 It is an alkyl group, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6. For example, R 3 は-OR 20 And R 20 is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, and optionally R 20 It is methyl.

[0110] One reason, R 3 は-OR 20 And R 20 is C 1-6 Alkoxy C 1-6 It is alkyl. For example, R 3 は-OR 20And R 20 It is 2-methoxyethyl.

[0111] One reason, R 3’ は-OR 20 And R 20 R is a hydroxyl protecting group. For example, R 3’ は-OR 20 And R 20This is BOC or Boc, MOM, MTM, t-butylthiomethyl, SMOM, BOM, PMBM, p-AOM, GUM, t-butoxymethyl, POM, siloxymethyl, MEM, 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, SEMOR, THP, 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, MTHP, 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl-S,S-dioxide, CTMP, 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-meth Nobenzofuran-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-trichloroethyl, 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'-dinitrobenzhydryl, 5-dibenzosberyl, 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(ruberinoyloxyphenyl)methyl, 4,4',4”-tris(benzoyloxyphenyl)methyl, 3-(imidazole-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-Benzodisulfuran-2-yl, benzisothiazolyl-S,S-dioxide, TMS, TES, TIPS, IPDMS, DEIPS, dimethyltexylsilyl, TBDMS, TBDPS, tripenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, DPMS, TBMPS, formate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate Triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (rebrinate), 4,4-(ethylenedithio)pentanoate (ruberinoyl dithioacetal), adamantoate, crotonate, 4-methoxycrotonate, bezoate, p-phenylbezoate, 2,4,6-trimethylbezoate (mesitoate), alkyl Methyl carbonate, Fmoc, alkyl ethyl carbonate, Troc, TMSEC, Psec, Peoc, alkyl isobutyl carbonate, alkyl vinyl carbonate, alkyl allyl 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-benzyl thiocarbonate, 4-ethoxy-1-naphthyl carbonate, methyl Dithiocarbonate, 2-iodobezoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)bezoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)bezoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, (E)-2-methyl-2-butanoate, o-(methoxyacyl)bezoate, α-naphthoate, nitrate, alkyl N,N,N',The hydroxyl protecting group is selected from the group consisting of N'-tetramethylphosphodiamide, alkyl N-phenylcarbamate, borate, dimethylphosphinothionel, alkyl 2,4-dinitrophenyl sulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate.

[0112] One reason, R 3’ は-OR 20 And R 20 The compound is TBDMS (or TBS), TBDPS, TMS, TES, TIPS, IPDMS, DEIPS, TBMPS, DPMS, dimethyltexylsilyl, tribenzylsilyl, tri-p-xylylsilyl, or triphenylsilyl, and optionally R 20 This is TBDMS (or TBS).

[0113] R 2 In the various embodiments described herein, R 2 is hydrogen, halogen, -OR 20 , or -OR 30 It is possible.

[0114] One reason, R 2 is hydrogen or halogen. For example, R 2 It is either H or F.

[0115] One reason, R 2 は-OR 20 And here, R 20 C is a hydrogen atom, a hydroxyl protecting group, or optionally substituted C 1-6 Alkyl, for example, C 1-6 Alkoxy C 1-6 Alkyl (e.g., 2-methoxyethyl) or N-(C) 1-6 Alkyl)aminocarbonyl C 1-6 It is an alkyl group (for example, 2-(N-methylamino)-2-oxoethyl). For example, R 2 は-OR 20 And R 20R is a hydrogen or hydroxyl protecting group. In other examples, R 2 は-OR 20 And here, R 20 C is replaced as desired. 1-6 It is alkyl.

[0116] One reason, R 2 は-OR 20 And R 20 Depending on the desired type, these are OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, NH(C1-C4)alkyl, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8)alkyl, O(C1-C8)alkyl (i.e., C1-C 8 alkoxy), O(C1-C8) haloalkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) p C is substituted with one, two, three, four, or five substituents independently selected from -NH2 or CH2-aryl-alkoxy. 1-6 It is an alkyl group, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6. For example, R 2 は-OR 20 And R 20 is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, and optionally R 20 It is methyl.

[0117] One reason, R 2 は-OR 20 And R20 Depending on the desired type, these are OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, NH(C1-C4)alkyl, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8)alkyl, O(C1-C8)alkyl (i.e., C1-C 8 alkoxy), O(C1-C8) haloalkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) p C is substituted with one, two, three, four, or five substituents independently selected from -NH2 or CH2-aryl-alkoxy. 1-6 Alkoxy C 1-6 It is an alkyl group, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6. For example, R 2 は-OR 20 And R 20 It is 2-methoxyethyl.

[0118] One reason, R 2’ は-OR 20 And R 20 R is a hydroxyl protecting group. For example, R 2’ は-OR 20 And R 20This is BOC or Boc, MOM, MTM, t-butylthiomethyl, SMOM, BOM, PMBM, p-AOM, GUM, t-butoxymethyl, POM, siloxymethyl, MEM, 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, SEMOR, THP, 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, MTHP, 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl-S,S-dioxide, CTMP, 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-meth Nobenzofuran-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-trichloroethyl, 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'-dinitrobenzhydryl, 5-dibenzosberyl, 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(ruberinoyloxyphenyl)methyl, 4,4',4”-tris(benzoyloxyphenyl)methyl, 3-(imidazole-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-Benzodisulfuran-2-yl, benzisothiazolyl-S,S-dioxide, TMS, TES, TIPS, IPDMS, DEIPS, dimethyltexylsilyl, TBDMS, TBDPS, tripenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, DPMS, TBMPS, formate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate Triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (rebrinate), 4,4-(ethylenedithio)pentanoate (ruberinoyl dithioacetal), adamantoate, crotonate, 4-methoxycrotonate, bezoate, p-phenylbezoate, 2,4,6-trimethylbezoate (mesitoate), alkyl Methyl carbonate, Fmoc, alkyl ethyl carbonate, Troc, TMSEC, Psec, Peoc, alkyl isobutyl carbonate, alkyl vinyl carbonate, alkyl allyl 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-benzyl thiocarbonate, 4-ethoxy-1-naphthyl carbonate, methyl Dithiocarbonate, 2-iodobezoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)bezoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)bezoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, (E)-2-methyl-2-butanoate, o-(methoxyacyl)bezoate, α-naphthoate, nitrate, alkyl N,N,N',The hydroxyl protecting group is selected from the group consisting of N'-tetramethylphosphodiamide, alkyl N-phenylcarbamate, borate, dimethylphosphinothionel, alkyl 2,4-dinitrophenyl sulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate.

[0119] One reason, R 2’ は-OR 20 And R 20 The compounds are TBDMS, TBDPS, TMS, TES, TIPS, IPDMS, DEIPS, TBMPS, DPMS, dimethyltexylsilyl, tribenzylsilyl, tri-p-xylylsilyl, or triphenylsilyl, and optionally R 20 This is TBDMS.

[0120] One reason, R 2 は-OR 30 And here, R 30 This can be a bond to hydrogen, a hydroxyl protecting group, a reactive phosphorus group (e.g., a phosphoramidite), a nucleoside or nucleotide, or an oligonucleotide. For example, R 2 は-OR 30 And R 30 is a hydrogen or hydroxyl protecting group.

[0121] One reason, R 2 は-OR 30 And R 30 R is a reactive phosphorus group. For example, R 30 is a phosphoramidite, H-phosphonate, alkyl-phosphonate, or phosphate triester. In one embodiment, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(ORP1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 These are independently OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, NH(C1-C4)alkyl, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8)alkyl, O(C1-C8)alkyl (i.e., C1-C8 Alkoxy), O(C1-C8)haloalkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) p C is substituted with one, two, three, four, or five substituents independently selected from -NH2 or CH2-aryl-alkoxy. 1-6 It is an alkyl group, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6; Each R P2Depending on the desired type, these are OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, NH(C1-C4)alkyl, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8)alkyl, O(C1-C8)alkyl (i.e., C1-C 8 alkoxy), O(C1-C8) haloalkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) p C is substituted with one, two, three, four, or five substituents independently selected from -NH2 or CH2-aryl-alkoxy. 1-6 It is an alkyl group, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6, and each R is optional. P2 These are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, preferably each R P2 It is isopropyl; or both R P2 These, together with the nitrogen atom to which they are bonded, form a 3- to 8-membered heterocycline which is optionally substituted; or R P1 and R P2 One of them unites with the atom to which they are bonded, forming a 4- to 8-membered heterocycline which is optionally substituted; and Each R P3 C is independent 1-30 Alkyl, C2-C 30 Alkenyl, or C2-C 30 Alkinyl (for example, C 1-10 Alkyl, C2-C 10Alkenyl, or C2-C 10 These are alkynyl compounds, each independently of choice being OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, NH(C1-C4)alkyl, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8)alkyl, O(C1-C8)alkyl(suna (C1-C8 alkoxy), O(C1-C8) haloalkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) p Substituted with one, two, three, four, or five substituents selected from -NH2 or CH2-aryl-alkoxy, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6, and each R optionally P3 The elements are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be optionally substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy.

[0122] One reason, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(RP2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 C is replaced with CN or -SC(O)Ph as desired. 1-6 It is alkyl; each R P2 C is independently replaced as desired. 1-6 It is alkyl; and each R P3 C is independently replaced as desired. 1-6 It is alkyl.

[0123] One reason, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 C is replaced with CN or -SC(O)Ph as desired.1-6 alkyl; each R P2 R is independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl; and each R P3 The elements are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be optionally substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy.

[0124] One reason, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 2-cyanoethyl (-CH2CH2CN); each R P2 C is independently replaced as desired. 1-6 It is alkyl; and each R P3 C is independently replaced as desired. 1-6 It is alkyl.

[0125] One reason, R 2 は-OR 30 And R 30 is -P(ORP1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 It is 2-cyanoethyl (-CH2CH2CN); each R P2 R is independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl; and each R P3 The elements are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be optionally substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy.

[0126] One reason, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2)2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 It is 2-cyanoethyl (-CH2CH2CN); each R P2 is independently isopropyl; and each R P3 The elements are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be optionally substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy.

[0127] One reason, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, where:R P1 2-cyanoethyl (-CH2CH2CN); each R P2 It is independently isopropyl.

[0128] One reason, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SRP1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 C is replaced with CN or -SC(O)Ph as desired. 1-6 Alkyl; both R P2 They form 3- to 8-membered heterocyclines which are optionally substituted with the nitrogen atom to which they are bonded; and each R P3 C is independently replaced as desired. 1-6 It is alkyl.

[0129] One reason, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 C is replaced with CN or -SC(O)Ph as desired. 1-6 Alkyl; both R P2 They form 3- to 8-membered heterocyclines which are optionally substituted with the nitrogen atom to which they are bonded; and each R P3The elements are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be optionally substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy.

[0130] One reason, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 It is 2-cyanoethyl (-CH2CH2CN); both R P2 They form 3- to 8-membered heterocyclines which are optionally substituted with the nitrogen atom to which they are bonded; and each R P3 C is independently replaced as desired. 1-6 It is alkyl.

[0131] One reason, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1)N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here: Each R P1 It is 2-cyanoethyl (-CH2CH2CN); both R P2 They form 3- to 8-membered heterocyclines which are optionally substituted with the nitrogen atom to which they are bonded; and each R P3 The elements are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be optionally substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy.

[0132] One reason, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )RP3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here:R P1 and R P2 One of them unites with the atom to which they are bonded, forming a 4- to 8-membered heterocycline which is optionally substituted; R P2 The other is C, which is independently substituted as desired. 1-6 It is alkyl; and each R P3 C is independently replaced as desired. 1-6 It is alkyl.

[0133] One reason, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here:R P1 and R P2 One of them unites with the atom to which they are bonded, forming a 4- to 8-membered heterocycline which is optionally substituted; R P2 The other is independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl; and each RP3 C is independently replaced as desired. 1-6 It is alkyl.

[0134] One reason, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here:R P1 and R P2 One of them unites with the atom to which they are bonded, forming a 4- to 8-membered heterocycline which is optionally substituted; R P2 The other is C, which is independently substituted as desired. 1-6 It is alkyl; and each R P3 The elements are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be optionally substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy.

[0135] One reason, R 2 は-OR 30 And R 30 is -P(OR P1 )N(R P2 )2, -P(SRP1 )N(R P2 )2, -P(O)(OR P1 )N(R P2 )2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 And here:R P1 and R P2 One of them unites with the atom to which they are bonded, forming a 4- to 8-membered heterocycline which is optionally substituted; R P2 The other is independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl; and each R P3 The elements are independently methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be optionally substituted with NH2, OH, C(O)NH2, COOH, halogen, SH, or C1-C6 alkoxy.

[0136] One reason, R 2 は-OR 30 And R 30 This is a bond to a nucleoside or nucleotide, or an oligonucleotide. When the bond is to a nucleoside, nucleotide, or oligonucleotide, the nucleotide bond between the compound of formulas I-II and the nucleoside, nucleotide, or oligonucleotide can be an unmodified (e.g., phosphodiester) nucleotide bond or a modified (e.g., phosphorothioate) nucleotide bond.

[0137] One reason, R 2 は-OR 30 And R 30 It is attached to the 5' position of a nucleoside, nucleotide, or oligonucleotide by an unmodified (e.g., phosphodiester) internucleotide bond (e.g., or a modified (e.g., phosphorothioate) internucleotide bond). For example, R 30 It binds to the 5' end of the oligonucleotide (e.g., 5'-OH). In one embodiment, R 30 It is attached to the 5' end (e.g., 5'-OH) of the oligonucleotide by an unmodified (e.g., phosphodiester) internucleotide bond. In another embodiment, R 30 It is attached to the 5' end (e.g., 5'-OH) of an oligonucleotide by a modified (e.g., phosphorothioate) internucleotide bond.

[0138] One reason, R 2’ は-OR 30 And R 30 R is a hydroxyl protecting group. For example, R 2’ は-OR 30 And R 30This is BOC or Boc, MOM, MTM, t-butylthiomethyl, SMOM, BOM, PMBM, p-AOM, GUM, t-butoxymethyl, POM, siloxymethyl, MEM, 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, SEMOR, THP, 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, MTHP, 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl-S,S-dioxide, CTMP, 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-meth Nobenzofuran-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-trichloroethyl, 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'-dinitrobenzhydryl, 5-dibenzosberyl, 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(ruberinoyloxyphenyl)methyl, 4,4',4”-tris(benzoyloxyphenyl)methyl, 3-(imidazole-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-Benzodisulfuran-2-yl, benzisothiazolyl-S,S-dioxide, TMS, TES, TIPS, IPDMS, DEIPS, dimethyltexylsilyl, TBDMS, TBDPS, tripenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, DPMS, TBMPS, formate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate Triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (rebrinate), 4,4-(ethylenedithio)pentanoate (ruberinoyl dithioacetal), adamantoate, crotonate, 4-methoxycrotonate, bezoate, p-phenylbezoate, 2,4,6-trimethylbezoate (mesitoate), alkyl Methyl carbonate, Fmoc, alkyl ethyl carbonate, Troc, TMSEC, Psec, Peoc, alkyl isobutyl carbonate, alkyl vinyl carbonate, alkyl allyl 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-benzyl thiocarbonate, 4-ethoxy-1-naphthyl carbonate, methyl Dithiocarbonate, 2-iodobezoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)bezoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)bezoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, (E)-2-methyl-2-butanoate, o-(methoxyacyl)bezoate, α-naphthoate, nitrate, alkyl N,N,N',The hydroxyl protecting group is selected from the group consisting of N'-tetramethylphosphodiamide, alkyl N-phenylcarbamate, borate, dimethylphosphinothionel, alkyl 2,4-dinitrophenyl sulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate.

[0139] One reason, R 2’ は-OR 30 And R 30 The compounds are TBDMS, TBDPS, TMS, TES, TIPS, IPDMS, DEIPS, TBMPS, DPMS, dimethyltexylsilyl, tribenzylsilyl, tri-p-xylylsilyl, or triphenylsilyl, and optionally R 30 It is TBDMS.

[0140] R V In one embodiment, at least one R V R is an independent hydroxyl protecting group. For example, at least one R V For example, both R VThese are independently ethyl, pivaloyloxymethyl (POM), BOC or Boc, MOM, MTM, t-butylthiomethyl, SMOM, BOM, PMBM, p-AOM, GUM, t-butoxymethyl, siloxymethyl, MEM, 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, SEMOR, THP, 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, MTHP, 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl-S,S-dioxide, CTMP, 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-to Rimethyl-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-trichloroethyl, 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'-dinitrobenzhydryl, 5-dibenzosberyl, 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(ruberinoyloxyphenyl)methyl, 4,4',4”-tris(benzoyloxyphenyl)methyl, 3-(imidazole-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-benzodisulfuran-2-yl, benzisothiazolyl-S,S-dioxide, TMS, TES, TIPS, IPDMS, DEIPS, dimethyltexylsilyl, TBDMS, TBDPS, tripenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, DPMS, TBMPS, formate, acetate, chloroacetate, diclom Loroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (rebrinate), 4,4-(ethylenedithio)pentanoate (ruberinoyl dithioacetal), adamantoate, crotonate, 4-methoxycrotonate, bezoate, p-phenylbezoate, 2,4,6-trimethylbezoate (mesitoate), alkyl Methyl carbonate, Fmoc, alkyl ethyl carbonate, Troc, TMSEC, Psec, Peoc, alkyl isobutyl carbonate, alkyl vinyl carbonate, alkyl allyl 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-benzyl thiocarbonate, 4-ethoxy-1-naphthyl carbonate, methyl Dithiocarbonate, 2-iodobezoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)bezoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)bezoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,The hydroxyl protecting group is selected from the group consisting of 1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, (E)-2-methyl-2-butanoate, o-(methoxyacyl)bezoate, α-naphthoate, nitrate, alkyl N,N,N',N'-tetramethylphosphodiamide, alkyl N-phenylcarbamate, borate, dimethylphosphinothionel, alkyl 2,4-dinitrophenylsulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate, and preferably the protecting group is POM or ethyl. Therefore, in some embodiments, at least one R, V For example, both R V is independently pivaloyloxymethyl (POM). In one other embodiment, at least one R V For example, both R V They are independent.

[0141] In one embodiment, at least one R V For example, both R V It is hydrogen independently.

[0142] B (nucleotide base) In some embodiments of the various aspects described herein, B is a nucleic acid base that is optionally modified. It should be noted that the nucleic acid base may be natural or unnatural. "Unnatural nucleic acid base" means a nucleic acid base other than adenine, guanine, cytosine, uracil, or thymine. Exemplary unnatural nucleic acid bases include inosine, xanthine, hypoxanthine, nubularin, isoguanidine, tubercidine, and substituted or modified analogs of adenine, guanine, cytosine, and uracil, e.g., 2-aminoadenine and adenine and guanine, other alkyl derivatives of 2-propyl and other alkyl derivatives of adenine and guanine, 5-halouracil and cytosine, 5-propynyluracil and cytosine, 6-azouracil, cytosine, and thymine, 5-uracil (pseudolacil), 4-thiouracil, 5-halouracil, and 5-(2-aminopropyl)uracil , 5-aminoallyluracil, 8-halo, amino, thiol, thioalkyl, hydroxyl and other 8-substituted adenines and guanines, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine, 5-substituted pyrimidine, 6-azapyrimidine and 2-aminopropyladenine, N-2, N-6 and O-6 substituted purines including 5-propynyluracil and 5-propynylcytosine, dihydrouracil, 3-deaza-5-azacytosine, 2-aminopurine, 5-alkyluracil, 7-alkylguanine, 5-alkylcytosine, 7-deazaadenine, N 6 ,N 6 -Dimethyladenine, 2,6-diaminopurine, 5-amino-allyl-uracil, N 3 -Methyluracil, substituted 1,2,4-triazole, 2-pyridinone, 5-nitroindole, 3-nitropyrrole, 5-methoxyuracil, uracil-5-oxyacetic acid, 5-methoxycarbonylmethyluracil, 5-methyl-2-thiouracil, 5-methoxycarbonylmethyl-2-thiouracil, 5-methylaminomethyl-2-thiouracil, 3-(3-amino-3 carboxypropyl)uracil, 3-methylcytosine, 5-methylcytosine, N 4 -acetylcytosine, 2-thiocytosine, N 6 -Methyladenine, N6 -Isopentyl adenine, 2-methylthio-N 6 -Isopentenyl adenine, N-methylguanine, or O-alkylated bases, but not limited thereto. Further purines and pyrimidines include those disclosed in U.S. Patent 3,687,808, Concise Encyclopedia of Polymer Science and Engineering, pages 858-859, Kroschwitz, JI, ed. John Wiley & Sons, 1990, and Englisch et al., Angewandte Chemie, International Edition, 1991, 30, 613, all of which are incorporated herein by reference.

[0143] In one embodiment, non-natural nucleic acid bases include inosine, xanthine, hypoxanthine, nubularin, isoguanidine, tubercidine, 2-(halo)adenine, 2-(alkyl)adenine, 2-(propyl)adenine, 2-(amino)adenine, 2-(aminoalkyl-l)adenine, 2-(aminopropyl)adenine, and 2-(methylthio)-N 6 -(isopentenyl)adenine, 7-(deaza)adenine, 8-(alkenyl)adenine, 8-(alkyl)adenine, 8-(alkynyl)adenine, 8-(amino)adenine, 8-(halo)adenine, 8-(hydroxyl)adenine, 8-(thioalkyl)adenine, 8-(thiol)adenine, N 6 -(isopentyl)adenine, N 6 -(methyl)adenine, N 6 ,N 6-(dimethyl)adenine, 2-(alkyl)guanine, 2-(propyl)guanine, 6-(alkyl)guanine, 6-(methyl)guanine, 7-(alkyl)guanine, 7-(methyl)guanine, 7-(deaza)guanine, 8-(alkyl)guanine, 8-(alkenyl)guanine, 8-(alkynyl)guanine, 8-(amino)guanine, 8-(halo)guanine, 8-(hydroxyl)guanine, 8-(thioalkyl)guanine Nin, 8-(thiol)guanine, N-(methyl)guanine, 2-(thio)cytosine, 3-(deaza)-5-(aza)cytosine, 3-(alkyl)cytosine, 3-(methyl)cytosine, 5-(alkyl)cytosine, 5-(alkynyl)cytosine, 5-(halo)cytosine, 5-(methyl)cytosine, 5-(propynyl)cytosine, 5-(propynyl)cytosine, 5-(trifluoromethyl)cytosine, 6-(azo)cytosine, N 4 -(acetyl)cytosine, 3-(3-amino-3-carboxypropyl)uracil, 2-(thio)uracil, 5-(methyl)-2-(thio)uracil, 5-(methylaminomethyl)-2-(thio)uracil, 4-(thio)uracil, 5-(methyl)-4-(thio)uracil, 5-(methylaminomethyl)-4-(thio)uracil, 5-(methyl)-2,4-(dithio)uracil, 5-(methylaminomethyl)-2,4-(dithio)uracil, 5-(2-aminopropyl)uracil, 5-(alkyl)uracil, 5-(alkynyl)uracil, 5-(allylamino)uracil, 5-(aminoallyl)uracil , 5-(aminoalkyl)uracil, 5-(guanidinium alkyl)uracil, 5-(1,3-diazole-1-alkyl)uracil, 5-(cyanoalkyl)uracil, 5-(dialkylaminoalkyl)uracil, 5-(dimethylaminoalkyl)uracil, 5-(halo)uracil, 5-(methoxy)uracil, uracil-5-oxyacetic acid, 5-(methoxycarbonylmethyl)-2-(thio)uracil, 5-(methoxycarbonyl-methyl)uracil, 5-(propynyl)uracil, 5-(propynyl)uracil, 5-(trifluoromethyl)uracil, 6-(azo)uracil, dihydrouracil, N 3-(methyl)uracil, 5-uracil (i.e., pseudouracil), 2-(thio)pseudracil, 4-(thio)pseudracil, 2,4-(dithio)pseudracil, 5-(alkyl)pseudracil, 5-(methyl)pseudracil, 5-(alkyl)-2-(thio)pseudracil, 5-(methyl)-2-(thio)pseudracil, 5-(alkyl)-4-(thio)pseudracil, 5-(methyl)-4-(thio)pseudracil, 5-(alkyl)-2,4-(dithio)pseudracil Douracil, 5-(methyl)-2,4-(dithio)pseudracil, 1-substituted pseudouracil, 1-substituted 2(thio)pseudracil, 1-substituted 4-(thio)pseudracil, 1-substituted 2,4-(dithio)pseudracil, 1-(aminocarbonylethylenyl)pseudracil, 1-(aminocarbonylethylenyl)-2(thio)pseudracil, 1-(aminocarbonylethylenyl)-4-(thio)pseudracil, 1-(aminocarbonylethylenyl)-2,4-(dithio)pseudracil Uracil, 1-(aminoalkylaminocarbonylethylenyl)-pseudracil, 1-(aminoalkylamino-carbonylethylenyl)-2(thio)-pseudracil, 1-(aminoalkylaminocarbonylethylenyl)-4-(thio)pseudracil, 1-(aminoalkylaminocarbonylethylenyl)-2,4-(dithio)pseudracil, 1,3-(diaza)-2-(oxo)-phenoxazine-1-yl, 1-(aza)-2-(thio)-3-(aza)-phenoxazine-1-yl, 1 ,3-(diaza)-2-(oxo)-phenothiazine-1-yl,1-(aza)-2-(thio)-3-(aza)-phenothiazine-1-yl,7-substituted 1,3-(diaza)-2-(oxo)-phenoxazine-1-yl,7-substituted 1-(aza)-2-(thio)-3-(aza)-phenoxazine-1-yl,7-substituted 1,3-(diaza)-2-(oxo)-phenothiazine-1-yl,7-substituted 1-(aza)-2-(thio)-3-(aza)-phenothiazine-1-yl,7-(aminoalkylhydroxyl)-1,3-(diaza)-2-(oxo)-phenoxadin-1-yl, 7-(aminoalkylhydroxyl)-1-(aza)-2-(thio)-3-(aza)-phenoxadin-1-yl, 7-(aminoalkylhydroxyl)-1,3-(diaza)-2-(oxo)-phenothiazine-1-yl, 7-(aminoalkylhydroxyl)-1-(aza)-2-(thio)-3-(aza)-phenothiazine-1-yl, 7-(guanidinium alkylhydroxyl)-1,3-(diaza)-2-(oxo)-phenoxadin-1-yl, 7-(guanidinium (Luylhydroxyl)-1-(aza)-2-(thio)-3-(aza)-phenoxazine-1-yl, 7-(guanidinium alkyl-hydroxyl)-1,3-(diaza)-2-(oxo)-phenothiazine-1-yl, 7-(guanidinium alkylhydroxyl)-1-(aza)-2-(thio)-3-(aza)-phenothiazine-1-yl, 1,3,5-(triaza)-2,6-(dioxa)-naphthalene, inosine, xanthine, hypoxanthine, nubularin, tubercidine, isoguanidine, inosinyl, 2-aza-inosinyl, 7-deaza - Inosinyl, nitroimidazolyl, nitropyrazolyl, nitrobenzimidazolyl, nitroindazolyl, aminoindolyl, pyrrolopyrimidinyl, 3-(methyl)isocarbostyrillyl, 5-(methyl)isocarbostyrillyl, 3-(methyl)-7-(propynyl)isocarbostyrillyl, 7-(aza)indolyl, 6-(methyl)-7-(aza)indolyl, imidizopyridinyl, 9-(methyl)-imidizopyridinyl, pyrrolopyridinyl, isocarbostyrillyl, 7-(propynyl)isocarbostyrillyl, propynyl-7-(aza)in Dylyl, 2,4,5-(trimethyl)phenyl, 4-(methyl)indolyl, 4,6-(dimethyl)indolyl, phenyl, naphthalenyl, anthracenyl, phenanthracenyl, pyrenyl, stilbenylon, tetracerenyl, pentaceryl, difluorotolyl, 4-(fluoro)-6-(methyl)benzimidazole, 4-(methyl)benzimidazole, 6-(azo)thymine, 2-pyridinone, 5-nitroindole, 3-nitropyrrole, 6-(aza)pyrimidine, 2-(amino)purine, 2,6-(diamino)purine, 5-substituted pyrimidine, N, 2- Substitution purine, N 6 - Substitution purine, O 6 -The group can be selected from substituted purines, substituted 1,2,4-triazoles, and any O-alkylated or N-alkylated derivatives thereof.

[0144] In one embodiment, the non-natural nucleic acid base is a modified nucleic acid base, i.e., the nucleic acid base includes the nucleic acid base modifications described herein, for example, the nucleic acid base is any substitution or modified analog of a natural nucleic acid base. Examples of nucleic acid base modifications include: C-5 pyrimidines having alkyl or aminoalkyl and other cationic groups such as guanidinium and amidine functional groups; purines having alkyl or aminoalkyl and other cationic groups such as guanidinium and amidine functional groups. 2 - and N 6 - Includes, but is not limited to, G-clamps, guanidinium G-clamps, and pseudouridines known in the art.

[0145] In some embodiment of the features, the non-natural nucleic acid base is a universal nucleic acid base. The universal nucleic acid base as used herein is any modified or unmodified natural or non-natural nucleic acid base that can base-pair with all adenine, cytosine, guanine, and uracil without substantially affecting the fusion behavior, recognition by intracellular enzymes, or activity of oligonucleotides containing the universal nucleic acid base. Some exemplary universal nucleic acid bases include, but are not limited to, 2,4-difluorotoluene, nitropyrrolyl, nitroindolyl, 8-aza-7-deazaadenine, 4-fluoro-6-methylbenzimidazole, 4-methylbenzimidazole, 3-methylisocarbostyryl, 5-methylisocarbostyryl, 3-methyl-7-propynylisocarbostyryl, 7-azaindolyl, 6-methyl-7-azaindolyl, imidizopyridinyl, 9-methylimidizopyridinyl, pyrrolopyridinyl, isocarbostyryl, 7-propynylisocarbostyryl, propynyl-7-azaindolyl, 2,4,5-trimethylphenyl, 4-methyllinolyl, 4,6-dimethylindolyl, phenyl, naphthalenyl, anthracenyl, phenanthracenyl, pyrenyl, stilbenyl, tetraceryl, pentaceryl, and their structural derivatives.

[0146] In some embodiments, natural or non-natural nucleic acid bases are protected nucleic acid bases. As used herein, “protected nucleic acid base” means a nucleic acid base that contains a nitrogen protecting group and / or an oxygen protecting group and / or a sulfur protecting group.

[0147] For example, a nucleic acid base is a pyrimidine modified at the C4 position. In another non-limiting example, a nucleic acid base is a pyrimidine modified at the C5 position.

[0148] In one embodiment, the nucleic acid base is a purine modified at the N2 position. In one embodiment, the nucleic acid base is N 6 It is a purine modified at the C6 position. For example, a nucleic acid base is a purine modified at the C6 position. In a non-restrictive example, a nucleic acid base is an N-7 deazapurine, optionally modified at the N7 position.

[0149] In one embodiment, the nucleic acid base is a modified, protected, or substituted analog of a nucleic acid base selected from adenine, cytosine, guanine, thymine, and uracil. For example, the nucleic acid base is uracil, adenine, guanine, or cytosine, each optionally independently containing a hydroxyl or amine protecting group.

[0150] double stranded RNA Those skilled in the art are well aware that double-stranded RNAs containing 19 to 24, and especially 21, base pairs are supported as being particularly effective in inducing RNA interference (RNAi). However, others have found that shorter or longer double-stranded oligonucleotides may be equally effective. Thus, in certain embodiments, the long double-stranded oligonucleotides described herein can induce RNA interference. In other words, the long double-stranded oligonucleotides described herein can intervene in RNA interference. The term "intervene in RNAi" as used herein means the ability to inhibit or reduce the expression of a target nucleic acid, e.g., target RNA, e.g., mRNA, in a sequence-specific manner.

[0151] Accordingly, in another embodiment, provided herein comprises a sense strand and an antisense strand that is substantially or 100% (e.g., exactly) complementary to the sense strand, wherein one of the sense strand or the antisense strand has the structure: [ka] [During the ceremony: X is either O or S; and Each R V [These are independently hydrogen or hydroxyl protecting groups.] It is a double-stranded RNA (dsRNA) with a 5' terminal modification that includes [specific component].

[0152] In one embodiment, the 5'-end modification of a sense strand or antisense strand (e.g., an antisense strand) is structural: [ka] Includes.

[0153] In one embodiment, X is O. In another embodiment, X is O, and each R V is hydrogen. In one embodiment, X is O, and each R V is ethyl. In one embodiment, X is O, and each R V It is pivaloyloxymethyl.

[0154] [ka] It should be noted that the sugar portion of a modified nucleotide, i.e., the nucleotide at the 5' end of the sense or antisense strand (5' terminal nucleotide), may contain a 5-membered or 6-membered ring. For example, the sugar portion of the modified nucleotide may be a furanose (e.g., ribofuranose, arabinofuranose, lyxofuranose, xylofuranose, librofuranose, or xylulofuranose, including its alpha and beta, D and L, deoxy, and modified derivatives) or a pyranose (e.g., glucopyranose, galactopyranose, mannopyranose, allopyranose, altropyranose, globyranose, idopyranose, and taropyranose, including its alpha and beta, D and L, deoxy, and modified derivatives).

[0155] in general, [ka] The modification replaces the CH2OH group in the sugar moiety of the 5' terminal nucleotide of the sense or antisense strand. For example, [ka] The modification replaces the 4'-CH2OH group of the furanose ring (e.g., ribofuranose, arabinofuranose, lyxofuranose, xylofuranose, librofuranose, or xylulofuranose, preferably ribofuranose) or the 5'-CH2OH group of the pyranose (e.g., glucopyranose, galactopyranose, mannopyranose, allopyranose, altropyranose, globyranose, idopyranose, or taropyranose, preferably glucopyranose, galactopyranose, or mannopyranose) at the 5' terminal nucleotide of the sense or antisense strand.

[0156] In one embodiment, the sense strand or antisense strand (e.g., the antisense strand) contains a compound of formula (I) described herein at its 5' end. For example, the 5' terminal nucleotide of the sense strand or antisense strand has the structure: [ka] [During the ceremony, Each R V These are independently hydrogen or a hydroxyl protecting group (e.g., ethyl or pivaloyloxymethyl ((CH3)3CC(O)OCH2-,POM); B is a modified nucleic acid base (e.g., uracil) as desired; R 2 and R 3 One of them is hydrogen, halogen, or -OR 20 And here: R 20 C is a hydrogen atom, a hydroxyl protecting group, or optionally substituted C 1-6 Alkyl (e.g., methyl, 2-methoxyethyl, 1,3-dimethoxyprop-2-yl, 2-(N-methylamino)-2-oxoethyl), optionally substituted C 2-6 alkenyl, or C substituted as desired. 2-6 Alkinyl (for example, propargyl); and R 2 and R 3 The other side is -OR 30 And here: R30 This refers to binding to an oligonucleotide (for example, an internucleotide bond that connects to a subsequent nucleotide of an oligonucleotide). It belongs to them.

[0157] In one embodiment, the 5' terminal nucleotide of the sense strand or antisense strand (e.g., the antisense strand) has the following structure: [ka] It has, and here R 2 and R 3 One of them is -OR 30 (For example, R 3 は-OR 30 ) and here:R 30 This refers to binding to an oligonucleotide (for example, an internucleotide bond that connects to a subsequent nucleotide of the oligonucleotide).

[0158] In another embodiment, the 5' terminal nucleotide of the sense strand or antisense strand (e.g., the antisense strand) has the following structure: [ka] It has, and here R 2 and R 3 One of them is -OR 30 (For example, R 3 は-OR 30 ) and here:R 30 This refers to binding to an oligonucleotide (for example, an internucleotide bond that connects to a subsequent nucleotide of the oligonucleotide).

[0159] In another embodiment, the 5' terminal nucleotide of the sense strand or antisense strand (e.g., the antisense strand) has the following structure: [ka] It has, and here R 2 and R 3 One of them is -OR 30 (For example, R3 は-OR 30 ) and here:R 30 This refers to binding to an oligonucleotide (for example, an internucleotide bond that connects to a subsequent nucleotide of the oligonucleotide).

[0160] In another embodiment, the 5' terminal nucleotide of the sense strand or antisense strand (e.g., the antisense strand) has the following structure: [ka] It has, and here R 2 and R 3 One of them is -OR 30 (For example, R 3 は-OR 30 ) and here:R 30 This refers to binding to an oligonucleotide (for example, an internucleotide bond that connects to a subsequent nucleotide of the oligonucleotide).

[0161] Preferably, the antisense strand of the dsRNA is an oligonucleotide as described herein.

[0162] The term "antisense strand" as used herein refers to an oligonucleotide that is substantially or 100% (e.g., exactly) complementary to the target nucleic acid of interest. For example, an antisense strand may be whole or partially complementary to the target nucleic acid of interest, such as messenger RNA, non-mRNA RNA sequences (e.g., microRNA, piwiRNA, tRNA, rRNA, and hnRNA), or coding or non-coding DNA sequences.

[0163] It should be noted that each strand of dsRNA can have a nucleotide length in the range of 12–40. For example, each strand can independently be 14–40 nucleotides long, 17–37 nucleotides long, 25–37 nucleotides long, 27–35 nucleotides long, 17–23 nucleotides long, 17–21 nucleotides long, 17–19 nucleotides long, 19–25 nucleotides long, 19–23 nucleotides long, 19–21 nucleotides long, 21–25 nucleotides long, 21–23 nucleotides long, 25–35 nucleotides long, 26–35 nucleotides long, 27–34 nucleotides long, 28–32 nucleotides long, or 29–31 nucleotides long. The sense strand and antisense strand may be of equal or different lengths, but are not limited to these. In one embodiment, the antisense strand is, for example, 1, 2, 3, 4, or 5 nucleotides longer than the sense strand.

[0164] In one embodiment, each of the sense and antisense strands is independently 15, 16, 17, 28, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides long. For example, each of the sense and antisense strands is independently 18, 19, 21, 22, 23, 24, or 25 nucleotides long. In one embodiment, each strand is independently 19, 20, 21, 22, or 23 nucleotides long. In one embodiment, one strand (e.g., the sense strand) is 18, 19, 20, 21, or 22 nucleotides long, and the other strand (e.g., the antisense strand) is 21, 22, 23, 24, or 25 nucleotides long.

[0165] The sense and antisense strands of a dsRNA molecule are complementary and can hybridize to form a double-stranded or double-stranded region. Therefore, a dsRNA molecule has a double-stranded or double-stranded region. The double-stranded region can be 17 to 25 nucleotide base pairs long. For example, dsRNA may have a double-stranded region of 17 to 24 nucleotide base pairs long. In one embodiment, dsRNA has a double-stranded region of 18, 19, 20, 21, 22, 22, 23, 24, or 25 nucleotide base pairs long. In another embodiment, dsRNA has a double-stranded region of 19, 20, 21, or 22 nucleotide base pairs long.

[0166] A dsRNA molecule has one or more overhang regions (i.e., single-stranded regions) and / or capping groups at the 3' end, 5' end, or both ends of the strand. The overhang may be 1 to 3 nucleotides long, e.g., 1, 2, or 3 nucleotides. The overhang may result from one strand being longer than the other, or from two strands of equal length being misaligned. The overhang may form a mismatch with the targeted sequence, be complementary to the targeted sequence, or be another sequence. The sense and antisense strands may also be linked, for example, by additional bases forming a hairpin, or by other non-base linkers. The overhang may be present at the 3' end of only one strand or at the 3' end of both strands.

[0167] In one embodiment, the dsRNA molecule contains a single overhang. For example, the dsRNA molecule has a single overhang, and the overhang does not exceed 1, 2, or 3 nucleotides in length. Preferably, the overhang is 2 nucleotides in length. In one embodiment, the overhang is located at the 3' end of the strand (e.g., the antisense strand). In one embodiment, the dsRNA contains a 2-nucleotide overhang at the 3' end of the strand (e.g., the antisense strand). For example, the overhang is located at the 3' end of the antisense strand. For example, the antisense contains a 1 or 2-nucleotide overhang at its 3' end.

[0168] dsRNA can also have blunt ends. For example, one end of a dsRNA may be blunt and the other end may have an overhang. The blunt end may be located at the 5' end of the antisense strand (or the 3' end of the sense strand), or vice versa. Generally, the antisense strand of a dsRNA has a nucleotide overhang at the 3' end and a blunt end at the 5' end. While not bound by theory, the asymmetric blunt end at the 5' end of the antisense strand and the 3' end overhang of the antisense strand are favorable for the guide strand to enter the RISC process. In one embodiment, the dsRNA has a 2-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[0169] In one other embodiment, the dsRNA molecule has two blunt ends, i.e., two dsRNA strands of equal length. In one embodiment, the antisense strand is 18–25 nucleotides long. In one embodiment, the antisense strand is 21–25, 19–25, 19–21, or 21–23 nucleotides long. In one particular embodiment, the antisense strand is 23 nucleotides long.

[0170] Similar to the antisense strand, the sense strand can be 18–25 nucleotides long in one embodiment. In one embodiment, the sense strand is 21–25, 19–25, 19–21, or 21–23 nucleotides long. In one embodiment, the sense strand is 21 nucleotides long.

[0171] In one embodiment, the sense strand is 21 nucleotides long and the antisense strand is 23 nucleotides long.

[0172] Nucleic acid modification The long double-stranded and single-stranded oligonucleotides described herein may contain one or more nucleic acid modifications. Exemplary nucleic acid modifications include, but are not limited to, nucleic acid base modifications, sugar modifications, sugar-linking modifications, conjugates (e.g., ligands), and any combination thereof. It should be noted that nucleic acid modifications may be present at any position on the long double-stranded and single-stranded oligonucleotides. Nucleic acid modifications may be present on only one strand of the dsRNA or on both strands. In some embodiments, the antisense strand alone contains at least one, e.g., two, three, four, five or more nucleic acid modifications. In some embodiments, the sense strand alone contains at least one, e.g., two, three, four, five or more nucleic acid modifications. In some embodiments, both strands independently contain at least one, e.g., two, three, four, five or more nucleic acid modifications.

[0173] The various embodiments described herein refer to specific positions on the chain, counting from the end of the chain. When the chain is single-stranded, for example, a long chain of oligonucleotides, the position is counted from the first nucleotide at the specified end. When the chain is a double-stranded molecule, for example, part of a long double-stranded oligonucleotide, the position can be counted from the first nucleotide at the specified end of the chain or from the first base pair nucleotide of the chain at the specified end. Preferably, the position is counted from the first nucleotide at the specified end of the chain.

[0174] Thermal destabilization modification In some embodiments of the aspects described herein, the dsRNA includes thermal destabilization modifications. "Thermal destabilization modifications" mean modifications that lower the overall melting temperature (Tm) of the dsRNA, preferably by 1, 2, 3, or 4 degrees compared to the Tm of dsRNA without such modifications. Exemplary thermal destabilization modifications are described herein below and include, but are not limited to, debasing modifications; mismatches with opposite nucleotides in opposing strands; and sugar modifications such as 2'-deoxy (i.e., 2'-H) modifications, acyclic nucleotides (e.g., unlocked nucleic acids (UNA) or glycol nucleic acids (GNA)), threose nucleic acids (TNA), nucleotides linked via the 2' position (i.e., nucleotides linked via a 2'-OH group at the 5' position of subsequent nucleotides (2'-5' RNA modifications)); Hyp spacer modifications; modified nucleotide-nucleotide bonds that reduce the thermal stability of the dsRNA double helix; or nucleic acid bases in which the W-CH bond of complementary bases in opposing strands is impaired.

[0175] In one embodiment, the dsRNA contains at least one independently selected thermal destabilization modification, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more. The thermal destabilization modifications can be located at any position on the dsRNA. Furthermore, all thermal destabilization modifications can be located on one or both strands of the dsRNA. In one embodiment, the antisense strand alone contains at least one thermal destabilization modification, for example, 2, 3, 4, or more. In one embodiment, the sense strand alone contains at least one thermal destabilization modification, for example, 2, 3, 4, or more. In one embodiment, both the sense and antisense strands contain at least one thermal destabilization modification, for example, 2, 3, 4, or more.

[0176] Thermal destabilization modifications can occur on any nucleotide of the sense or antisense strand. For example, thermal destabilization modifications can occur on all nucleotides of the sense and / or antisense strand; each thermal destabilization modification can occur in an alternating pattern on the sense or antisense strand; or both the sense and antisense strands may contain an alternating pattern of thermal destabilization modifications. The alternating pattern of thermal destabilization modifications on the sense strand may be identical or different to that on the antisense strand, and the alternating pattern of thermal destabilization modifications on the sense strand may be shifted relative to the alternating pattern of thermal destabilization modifications on the antisense strand.

[0177] In one embodiment, the thermal destabilization modification is located at positions 2, 3, 4, 5, 6, 7, 8, or 9, or preferably positions 4, 5, 6, 7, or 8, counting from the 5' end of the antisense chain. In another embodiment, the thermal destabilization modification is located at positions 2, 3, 4, 5, or 9 from the 5' end of the antisense chain. In a particular embodiment, the thermal destabilization modification is located at position 6, 7, or 8 from the 5' end of the antisense chain.

[0178] In one embodiment, only the antisense chain includes a thermal destabilization modification. For example, the antisense chain includes a thermal destabilization modification, which is located at position 4, 5, 6, 7, or 8, counting from the 5' end of the antisense chain; preferably the thermal destabilization modification is located at position 5, 6, 7, or 8; more preferably the thermal destabilization modification is located at position 6, 7, or 8. In one embodiment, the antisense chain includes a thermal destabilization modification, which is located at position 7, counting from the 5' end of the antisense chain.

[0179] Similar to antisense chains, the thermal destabilization modification may be located at position 2, 3, 4, 5, 6, 7, 8, or 9, or preferably position 4, 5, 6, 7, or 8, counting from the 5' end of the long ssNA. In one embodiment, the thermal destabilization modification is located at position 2, 3, 4, 5, or 9 from the 5' end of the long ssNA. In another embodiment, the thermal destabilization modification is located at position 6, 7, or 8 from the 5' end of the long ssNA. In one particular embodiment, the thermal destabilization modification is located at position 7 from the 5' end of the long ssNA.

[0180] Heat stabilization modification In some embodiments, the dsRNA includes a thermal destabilization modification. “Thermal stabilization modification” means a modification that increases the overall melting temperature (Tm) of the dsRNA, preferably by 1, 2, 3, or 4 degrees above the Tm of dsRNA without such modification. Exemplary thermal destabilization modifications are described herein and may include, but are not limited to, 2'-fluoronucleotides (2'-F modifications), cross-linked nucleic acids (BNAs), e.g., loc nucleic acids (LNAs), and cyclohexene nucleic acids (CeNAs). In some preferred embodiments, the thermal stabilization modification is a 2'-fluoronucleotide. An example of a thermal stabilization modification is described herein. Further exemplary debasic nucleotides, acyclic nucleotide modifications (including UNAs and GNAs), and mismatch modifications are detailed in WO2011 / 133876 and WO2019222479, both of which are incorporated herein by reference in their entirety.

[0181] In some embodiments, a dsRNA may contain at least two, for example, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen or more heat-stabilizing (e.g., 2'-F) modifications. However, all heat-stabilizing (e.g., 2'-F) modifications may be present on one or both strands of the dsRNA. In some embodiments, the sense strand may contain at least one, for example, two, three, four or more heat-stabilizing (e.g., 2'-F) modifications. In some embodiments, the antisense strand may contain at least one, for example, two, three, four or more heat-stabilizing (e.g., 2'-F) modifications. In some embodiments, both the sense and antisense strands may contain at least one, for example, two, three, four or more heat-stabilizing (e.g., 2'-F) modifications. Heat-stabilizing (e.g., 2'-F) modifications may occur on any nucleotide of the sense or antisense strand. For example, thermal stabilization (e.g., 2'-F) modifications may occur on all nucleotides of the sense and / or antisense strands; each thermal stabilization (e.g., 2'-F) modification may occur in an alternating pattern on the sense or antisense strand; or both the sense and antisense strands may contain thermal stabilization (e.g., 2'-F) modifications as thermal destabilization modifications. The alternating pattern of thermal stabilization (e.g., 2'-F) modifications on the sense strand may be identical or different to that on the antisense strand, and the alternating pattern of thermal stabilization (e.g., 2'-F) modifications on the sense strand may be shifted relative to the alternating pattern of thermal stabilization (e.g., 2'-F) modifications on the antisense strand.

[0182] In one embodiment, the sense strand of dsRNA contains at least one, for example, two, three, four, five, six, seven, eight, nine, ten or more heat-stabilizing (e.g., 2'-F) modifications. In another embodiment, the sense strand contains two, three, four, or five heat-stabilizing (e.g., 2'-F) modifications. For example, the sense strand contains three or four heat-stabilizing (e.g., 2'-F) modifications. The heat-stabilizing (e.g., 2'-F) modifications on the sense strand may be located at any position, but are not limited to these. In one embodiment, the sense strand contains at least three heat-stabilizing (e.g., 2'-F) modifications. For example, the sense contains heat-stabilizing (e.g., 2'-F) modifications at at least positions 7, 10, and 11, counting from the 5' end of the sense strand. In yet another embodiment, the sense strand contains at least four heat-stabilizing (e.g., 2'-F) modifications. For example, the sense chain contains thermally stabilized (e.g., 2'-F) modifications at at least the 7th, 9th, 10th, and 11th positions, counting from the 5' end of the sense chain.

[0183] In one embodiment, the sense chain includes thermally stabilized (e.g., 2'-F) modifications at positions opposite or complementary to positions 11, 12, and 15 of the antisense chain, counting from the 5' end of the antisense chain. In another embodiment, the sense chain includes thermally stabilized (e.g., 2'-F) modifications at positions opposite or complementary to positions 11, 12, 13, and 15 of the antisense chain, counting from the 5' end of the antisense chain. In one embodiment, the sense chain includes two, three, or four blocks of thermally stabilized (e.g., 2'-F) modifications.

[0184] In one embodiment, the sense chain includes thermally stabilized (e.g., 2'-F) modifications at at least positions 7, 9, and 11 counting from the 5' end of the sense chain, and the antisense chain includes thermally stabilized (e.g., 2'-F) modifications at at least positions 2, 14, and 16 counting from the 5' end of the antisense chain. In another embodiment, the sense chain includes thermally stabilized (e.g., 2'-F) modifications at at least positions 7, 9, and 11 counting from the 5' end of the sense chain, and the antisense chain includes thermally stabilized (e.g., 2'-F) modifications at at least positions 2, 6, 9, 14, and 16 counting from the 5' end of the antisense chain. In yet another embodiment, the sense chain includes a thermally stabilized (e.g., 2'-F) modification at at least the 7th, 9th, and 11th positions counting from the 5' end of the sense chain, and the antisense chain includes a thermally stabilized (e.g., 2'-F) modification at at least the 2nd, 6th, 8th, 9th, 14th, and 16th positions counting from the 5' end of the antisense chain.

[0185] In one embodiment, the sense chain includes thermally stabilized (e.g., 2'-F) modifications at at least positions 7, 9, 10, and 11 counting from the 5' end of the sense chain, and the antisense chain includes thermally stabilized (e.g., 2'-F) modifications at at least positions 2, 14, and 16 counting from the 5' end of the antisense chain. In another embodiment, the sense chain includes thermally stabilized (e.g., 2'-F) modifications at at least positions 7, 9, 10, and 11 counting from the 5' end of the sense chain, and the antisense chain includes thermally stabilized (e.g., 2'-F) modifications at at least positions 2, 6, 9, 14, and 16 counting from the 5' end of the antisense chain. In yet another embodiment, the sense chain includes thermally stabilized (e.g., 2'-F) modifications at at least positions 7, 9, 10, and 11 counting from the 5' end of the sense chain, and the antisense chain includes thermally stabilized (e.g., 2'-F) modifications at at least positions 2, 6, 8, 9, 14, and 16 counting from the 5' end of the antisense chain.

[0186] In one embodiment, the sense chain does not contain thermal stabilization (e.g., 2'-F) modifications in a position opposite or complementary to the thermal destabilization modifications of the antisense chain's double helix.

[0187] The antisense strand molecule of dsRNA may contain at least one, for example, two, three, four, five, six, seven, eight, nine, ten, or more heat-stabilizing (e.g., 2'-F) modifications. In one embodiment, the antisense strand contains two, three, four, five, or six heat-stabilizing (e.g., 2'-F) modifications. The heat-stabilizing (e.g., 2'-F) modifications on the antisense strand may be located at any position, but are not limited. In one embodiment, the antisense strand contains at least three heat-stabilizing (e.g., 2'-F) modifications. For example, the antisense strand contains heat-stabilizing (e.g., 2'-F) modifications at at least positions 2, 14, and 16 counting from the 5' end of the antisense strand. In another embodiment, the antisense contains at least four heat-stabilizing (e.g., 2'-F) modifications. For example, the antisense chain includes thermally stabilized (e.g., 2'-F) modifications at at least positions 2, 6, 14, and 16, counting from the 5' end of the antisense chain. In a further embodiment, the antisense chain includes at least five thermally stabilized (e.g., 2'-F) modifications. For example, the antisense chain includes thermally stabilized (e.g., 2'-F) modifications at at least positions 2, 6, 9, 14, and 16, counting from the 5' end of the antisense chain. In a further embodiment, the antisense chain includes at least six thermally stabilized (e.g., 2'-F) modifications. For example, the antisense chain includes thermally stabilized (e.g., 2'-F) modifications at at least positions 2, 6, 8, 9, 14, and 16, counting from the 5' end of the antisense chain.

[0188] In one embodiment, the antisense strand includes at least one thermally stabilizing (e.g., 2'-F) modification adjacent to a stabilizing / destabilizing modification. For example, the thermally stabilizing (e.g., 2'-F) modification may be at the 5' or 3' end of the thermally destabilizing modification, i.e., at the -1 or +1 position from the position of the thermally destabilizing modification. In one embodiment, the antisense strand includes thermally stabilizing (e.g., 2'-F) modifications at the 5' and 3' ends of the thermally destabilizing modification, i.e., at the -1 and +1 positions from the position of the destabilizing modification.

[0189] In one embodiment, the antisense chain includes at least two stabilizing modifications at the 3' end of the destabilizing modification, i.e., at the +1 and +2 positions from the position of the destabilizing modification.

[0190] In one embodiment, the sense chain does not contain thermal stabilization (e.g., 2'-F) modifications in a position opposite or complementary to the thermal destabilization modifications of the antisense chain's double helix.

[0191] 2'-OMenucleotide In one embodiment, the dsRNA described herein may contain at least one, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more 2'-OMe nucleotides. Not limited to these, all 2'-OMe nucleotides may be present on one or both strands of the dsRNA. In one embodiment, both the sense strand and the antisense strand contain at least one 2'-OMe nucleotide. 2'-OMe modifications can occur on any nucleotide of the sense strand or antisense strand. For example, 2'-OMe modifications may occur on all nucleotides of the sense strand and / or antisense strand; they may occur in an alternating pattern on each 2'-OMe modified sense strand or antisense strand; or both the sense strand and antisense strand may contain 2'-OMe modifications via thermal destabilization modifications. The alternating pattern of 2'-OMe modifications on the sense chain may be the same as or different from that on the antisense chain, and the alternating pattern of 2'-OMe modifications on the sense chain may be shifted relative to the alternating pattern of 2'-OMe modifications on the antisense chain.

[0192] The antisense strand molecule of dsRNA may contain at least one, for example, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, or more 2'-OMe modifications. The 2'-OMe modifications may be located at any position on the antisense strand, but are not limited to these. In one embodiment, each nucleotide is independently a 2'-O-methylnucleotide, except for any other specific modifications on the antisense strand (e.g., thermal destabilization modifications, thermal stabilization modifications, and / or 2'-deoxy(2'-H) modifications).

[0193] Similar to the antisense strand, the sense strand molecule of dsRNA may contain at least one, for example, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, or more 2'-OMe modifications. The 2'-OMe modifications on the sense strand may be located at any position, but are not limited to these. In one embodiment, each nucleotide is independently a 2'-O-methylnucleotide, with the exception of any other specific modifications on the sense strand (e.g., thermal stabilization modifications, lipophilic modifications, inverted nucleotides, thermal destabilization modifications, and / or 2'-deoxy(2'-H) modifications).

[0194] 2'-deoxy(2'-H)nucleotide In one embodiment, the dsRNA described herein may contain 2'-deoxyribonucleotides, i.e., 2'-H nucleotides. For example, the long double-stranded and single-stranded oligonucleotides described herein may contain at least one (e.g., one, two, three, four, five or more) 2'-deoxyribonucleotides.

[0195] The 2'-deoxynucleotide may be located at any position on the sense strand or antisense strand. Furthermore, all 2'-deoxynucleotides may be present on one or both strands of the dsRNA.

[0196] In one embodiment, the sense strand contains one, two, three, four, five or more 2'-deoxynucleotides. For example, the sense strand contains a 2'-deoxynucleotide at any one of the positions 7, 9, and 11, counting from the 5' end of the strand. In another embodiment, the sense strand contains a 2'-deoxynucleotide at at least position 9, counting from the 5' end of the strand. For example, the sense strand contains a 2'-deoxynucleotide at at least positions 7 and 9, counting from the 5' end of the strand. In other non-limiting examples, the sense strand contains a 2'-deoxynucleotide at at least positions 9 and 11, counting from the 5' end of the strand.

[0197] In one embodiment, the antisense strand contains one, two, three, four, five, six, seven, eight or more 2'-deoxynucleotides. For example, the antisense strand contains a 2'-deoxynucleotide at any one of the following positions, counting from the 5' end of the strand: 2, 5, 7, 12, 14, and 16. In one embodiment, the antisense strand contains a 2'-deoxynucleotide at at least the 5th position, counting from the 5' end of the strand. For example, the antisense strand contains a 2'-deoxynucleotide at at least the 2nd, 5th, and 9th positions, counting from the 5' end of the strand. In other non-limiting examples, the antisense strand contains a 2'-deoxynucleotide at at least the 2nd, 5th, 7th, and 12th positions, counting from the 5' end of the strand. In other non-limiting examples, the antisense strand contains 2'-deoxynucleotides at positions 2, 5, 7, 12, 14, and 16, counting from the 5' end of the strand.

[0198] Lipophilic modification In one embodiment, the dsRNA described herein may include lipophilic modifications. For example, the long double-stranded and single-stranded oligonucleotides described herein may include at least one (e.g., one, two, three, four, five or more) lipophilic modifications. An exemplary lipophilic modification is a nucleotide having a modification at a lipophilic group, for example, a lipophilic group at the 2' position (e.g., C 10 -C 30 Alkyl, or C 10 -C30 Alkenyl group, for example, C 16 Alkyl, C 16 Alkenil, C 18 Alkyl, C 18 Alkenil, C 20 Alkyl, C 20 Alkenil, C 22 Alkyl, C 22 Alkenil, C 24 Alkyl, C 24 Alkenil; C 15 Alkyl, C 15 Alkenil, C 17 Alkyl, C 17 Alkenil, C 19 Alkyl, C 19 Alkenil, C 21 Alkyl, C 21 Alkenil, C 23 Alkyl, or C 23 These may include nucleotides containing alkenyls. Some exemplary lipophilic nucleotides include, but are not limited to, 2'-O-hexadecyl-modified nucleotides (Nhd), 2'-O-docosanyl-modified nucleotides (Nda), 2'-O-(omega-hydroxy-hexadecyl)-modified nucleotides (NhdOH), and 2'-O-(omega-hydroxy-docosanyl)-modified nucleotides (NdaOH).

[0199] Lipophilic modifications may be located at any position on the sense strand or antisense strand. Furthermore, all lipophilic modifications may be present on one or both strands of the dsRNA. In one embodiment, only the sense strand contains lipophilic modifications. For example, the sense strand contains a lipophilic modification at any one of the following positions, counting from the 5' end: position 1, 2, 3, 4, 5, 6, 7, 8, 13, 14, 15, 16, 17, or 18. In another embodiment, the sense strand contains a lipophilic modification at any one of the following positions, counting from the 5' end: position 4, 5, 6, 7, 8, 13, 14, 15, 16, 17, or 18.

[0200] In one embodiment, each residue in the sense and antisense strands is independently modified with 2'-O-methyl, 2'-fluoro, 2'-deoxy, LNA, HNA, CeNA, 2'-methoxyethyl, 2'-O-allyl, or 2'-C-allyl, 2'-deoxy. The strands may contain more than one modification. In one embodiment, each residue in the sense and antisense strands is independently modified with 2'-O-methyl or 2'-fluoro. It should be understood that these modifications are in addition to any other identified modifications of the dsRNA molecule (e.g., at least one double-strand thermal destabilization modification present in the antisense strand).

[0201] In one embodiment, at least two distinct modifications are typically present in the sense and antisense strands. These two modifications may be 2'-deoxy, 2'-O-methyl, or 2'-fluoro modifications, or thermal destabilization modifications. In one embodiment, the sense and antisense strands each contain two distinct modified nucleotides selected from 2'-O-methyl or 2'-deoxy. In one embodiment, each residue in the sense and antisense strands is independently modified with a 2'-O-methyl nucleotide, a 2'-deoxy nucleotide, a 2'-deoxy-2'-fluoro nucleotide, a 2'-ON-methylacetamide (2'-O-NMA) nucleotide, a 2'-O-dimethylaminoethoxyethyl (2'-O-DMAEOE) nucleotide, a 2'-O-aminopropyl (2'-O-AP) nucleotide, or a 2'-ara-F nucleotide. For example, each residue in the sense and antisense strands is independently modified with a 2'-O-methyl nucleotide, a 2'-deoxy nucleotide, or a 2'-deoxy-2'-fluoro nucleotide. Similarly, it should be understood that these modifications are in addition to any double-chain thermal destabilization modifications present in the antisense chain.

[0202] In one embodiment, the antisense chain includes at least one thermal destabilization modification, and the remaining nucleotides are independently 2'-O-methylnucleotide, 2'-deoxynucleotide, 2'-deoxy-2'-fluoronucleotide, 2'-ON-methylacetamide (2'-O-NMA) nucleotide, 2'-O-dimethylaminoethoxyethyl (2'-O-DMAEOE) nucleotide, 2'-O-aminopropyl (2'-O-AP) nucleotide, or 2'-ara-F nucleotide. For example, the antisense chain includes a thermal destabilization modification, and the remaining nucleotides are independently 2'-O-methylnucleotide, 2'-deoxynucleotide, or 2'-deoxy-2'-fluoronucleotide. In one embodiment, the antisense chain comprises (i) a thermal destabilization modification at the 5th, 6th, 7th, or 8th position counted from the 5' end of the antisense chain; (ii) at least two, e.g., three, four, five, or six 2'-fluoronucleotides; and (iii) the remaining nucleotides, independently being 2'-O-methylnucleotides or 2'-deoxynucleotides.

[0203] In one embodiment, each nucleotide in the sense strand is independently a 2'-O-methylnucleotide, a 2'-deoxynucleotide, a 2'-deoxy-2'-fluoro(2'-F)nucleotide, a 2'-ON-methylacetamide(2'-O-NMA)nucleotide, a 2'-O-dimethylaminoethoxyethyl(2'-O-DMAEOE)nucleotide, a 2'-O-aminopropyl(2'-O-AP)nucleotide, or a 2'-ara-F nucleotide. For example, each nucleotide in the sense strand is independently a 2'-O-methylnucleotide, a 2'-deoxynucleotide, or a 2'-deoxy-2'-fluoronucleotide. In one embodiment, the sense strand contains at least two, for example, three, four, five, or six 2'-fluoronucleotides, and the remaining nucleotides are independently 2'-O-methylnucleotides or 2'-deoxynucleotides.

[0204] In one embodiment, at least one of the first one, two, three, four, or five base pairs from the 5' end of the antisense strand within the double-stranded region of dsRNA may be independently selected from the group consisting of A:U, G:U, I:C, and mismatch pairs, e.g., non-canonical or non-canonical pairings or pairs containing universal bases, in order to facilitate the dissociation of the antisense strand at the 5' end of the double helix. In one embodiment, the nucleotide at position 1 from the 5' end of the antisense strand within the double-stranded region is selected from the group consisting of A, dA, dU, U, and dT. Alternatively, at least one of the first one, two, or three base pairs from the 5' end of the antisense strand within the double-stranded region is an AU base pair. For example, the first base pair from the 5' end of the antisense strand within the double-stranded region is an A:U base pair. It should be noted that the sense strand or antisense strand may contain an adenosine (A) nucleotide.

[0205] Inter-modified nucleotide bonding The dsRNAs described herein may contain at least one, for example, two, three, four, five, six, seven, eight, nine, ten, or more modified nucleoside bonds. The term "nucleoside bond" as used herein refers to a covalent bond between adjacent nucleosides.Exemplary modified nucleoside bonds include phosphodithioates, phosphorothioates (R, S, or racemic), phosphorodithioates, methylene methylimino (MMI, 3'-CH2-N(CH3)-O-5'), phosphotriesters, alkylphosphonates (e.g., methylphosphonate), phosphoramidates, methylene methylimino (-CH2-N(CH3)-O-CH2-), thiodiesters (-OC(O)-S-), thionocarbamates (-OC(O)(NH)-S-), and siloxanes (-O-Si( H)2-O- and dialkylsiloxanes), N,N'-dimethylhydrazine (-CH2-N(CH3)-N(CH3)-), amide-3 (3'-CH2-C(=O)-N(H)-5'), amide-4 (3'-CH2-N(H)-C(=O)-5'), hydroxylamino, siloxane (dialkylsiloxane), carboxamide, carbonate, carboxymethyl, carbamate, carboxylate ester, thioether, ethylene oxide linker, sulfide, sulfonate, sulfonamide, Sulfonate esters, thioformacetal (3'-S-CH2-O-5'), formacetal (3'-O-CH2-O-5'), oxime, methylene imino, methylene carbonylamino, methylene hydrazo, methylene dimethyl hydrazo, methylene oxymethyl imino, ether (C3'-O-C5'), thioether (C3'-S-C5'), thioacetamide (C3'-N(H)-C(=O)-CH2-S-C5', C3'-OP(O)-O-SS-C5'), C3'-CH2-NH-NH-C5 The modified internucleoside bonds include, but are not limited to, ', 3'-NHP(O)(OCH3)-O-5', 3'-NHP(O)(OCH3)-O-5', imidophosphoramidates ("imidp"), 2'→5' nucleoside bonds, 2'→3 nucleoside bonds, 3'→3 nucleoside bonds, and 5'→5' nucleoside bonds, and the modified internucleoside bonds may optionally be phosphorothioates, methylphosphonates, imidp or MMI, and more preferably phosphorothioates (PS).

[0206] Modified nucleotide-linking modifications can occur on any nucleotide at any position on the sense strand, the antisense strand, or both. For example, nucleotide-linking modifications can occur on all nucleotides of the sense strand and / or antisense strand; each nucleotide-linking modification can occur in an alternating pattern on the sense strand or antisense strand; or the sense strand or antisense strand contains alternating patterns of both nucleotide-linking modifications. The alternating pattern of nucleotide-linking modifications on the sense strand may be identical or different to that on the antisense strand, and the alternating pattern of nucleotide-linking modifications on the sense strand may be shifted relative to the alternating pattern of nucleotide-linking modifications on the antisense strand.

[0207] In one embodiment, the dsRNA includes modified nucleoside (e.g., phosphorothioate or methylphosphonate) bonds in the overhang region. For example, the overhang region includes two nucleotides having modified nucleoside (e.g., phosphorothioate or methylphosphonate) bonds between the two nucleotides. The nucleotide bond modification may also link the overhang nucleotide to a terminal pair nucleotide within the double-stranded region. For example, at least two, three, four, or all of the overhang nucleotides may be linked via modified nucleoside (e.g., phosphorothioate or methylphosphonate) bonds, and optionally there may be further modified nucleoside (e.g., phosphorothioate or methylphosphonate) bonds linking the overhang nucleotide to the adjacent pair nucleotide. For example, there may be at least one modified nucleoside bond (e.g., between phosphorothioate or methylphosphonate nucleotides) between the three terminal nucleotides, where two of the three nucleotides are overhang nucleotides and the third is a paired nucleotide adjacent to the overhang nucleotide. Preferably, these three terminal nucleotides may be at the 3' end of the antisense strand.

[0208] Regarding the position of internucleotide bonds, the indicated position refers to the internucleotide bond that connects the nucleotide at that position to the nucleotide of the 1-position granule from that position. In other words, the internucleotide bond at position N means that it is between nucleotides N and N+1. Therefore, counting from the 5' end, the internucleotide bond at position 1 means that the linker is between the nucleotides at positions 1 and 2, counting from the 5' end.

[0209] In one embodiment, the sense strand includes 1 to 5 modified nucleoside (e.g., phosphorothioate or methylphosphonate nucleotide) bonds within positions 1 to 5, counting from the 5' end of the sense strand, and 1 to 5 modified nucleoside (e.g., phosphorothioate or methylphosphonate nucleotide) bonds within positions 1 to 5, counting from the 5' end of the sense strand. For example, the sense strand contains modified nucleoside (e.g., phosphorothioate or methylphosphonate) bonds between the nucleotides at positions 1 and 2, and between the nucleotides at positions 2 and 3, counting from the 5' end of the sense strand, and the sense strand further contains modified nucleoside (e.g., phosphorothioate or methylphosphonate) bonds between the nucleotides at positions 1 and 2, and between the nucleotides at positions 2 and 3, counting from the 3' end of the sense strand.

[0210] In one embodiment, the antisense chain includes 1 to 5 modified nucleoside (e.g., phosphorothioate or methylphosphonate) bonds within the 1 to 5 positions, counting from the 5' end of the antisense chain, and 1 to 5 modified nucleoside (e.g., phosphorothioate or methylphosphonate) bonds within the 1 to 5 positions, counting from the 5' end of the antisense chain. For example, the antisense strand includes modified nucleoside (e.g., phosphorothioate or methylphosphonate) bonds between the nucleotides at positions 1 and 2, and between the nucleotides at positions 2 and 3, counting from the 5' end of the antisense strand, and further includes modified nucleoside (e.g., phosphorothioate or methylphosphonate) bonds between the nucleotides at positions 1 and 2, and between the nucleotides at positions 2 and 3, counting from the 3' end of the antisense strand.

[0211] In one embodiment, the sense strand includes modified nucleoside (e.g., phosphorothioate or methylphosphonate) bonds at positions 1 and 2, counting from the 5' end of the sense strand, and modified nucleoside (e.g., phosphorothioate or methylphosphonate) bonds at positions 1 and 2, counting from the 3' end of the sense strand; and the antisense strand includes modified nucleoside (e.g., phosphorothioate or methylphosphonate) bonds at positions 1 and 2, counting from the 5' end of the antisense strand, and modified nucleoside (e.g., phosphorothioate or methylphosphonate) bonds at positions 1 and 2, counting from the 3' end of the antisense strand.

[0212] Overhang modification The nucleotides in the overhang region of a dsRNA molecule can be modified or unmodified nucleotides, each independently including, but not limited to, 2'-sugar modifications such as 2'-fluoro, 2'-O-methyl, thymidine (T), 2'-O-methoxyethyl-5-methyluridine, 2'-O-methoxyethyladenosine, 2'-O-methoxyethyl-5-methylcytidine, GNA (glycol nucleic acid), SNA (serinol nucleic acid), TNA (threose nucleic acid), and any combination thereof. For example, TT (or UU) can be an overhang sequence at either end of either strand. The sense strand, antisense strand, or the 5'- or 3'-overhangs of both strands of a dsRNA molecule can be phosphorylated. In one embodiment, the overhang region comprises two nucleotides having a phosphorothioate internucleotide bond between them, where the two nucleotides in the overhang region may be identical or different.

[0213] 5'-modified [ka] The 5' end of a dsRNA strand lacking modification can also be modified. Exemplary modifications of the 5' end include, but are not limited to, 5'-morpholinonucleotides (e.g., nucleotides in which the 5'-OH group is replaced by a morpholino group), 5'-dimethylaminonucleotides (e.g., nucleotides in which the 5'-OH group is replaced by a dimethylamino group), 5'-deoxynucleotides, inverted nucleotides (i.e., nucleotides attached to the rest of the strand via a 5'→5' bond), inverted debasalized nucleotides (e.g., debasalized nucleotides attached via a 5'→5' bond), or inverted debasalized locked nucleic acid modifications (i.e., LNAs lacking a nucleic acid base and attached via a 5'→5' bond).

[0214] In one embodiment, the sense strand of dsRNA contains a 5'-morpholinonucleotide, a 5'-dimethylaminonucleotide, a 5'-deoxynucleotide, an inverted nucleotide, an inverted debasalized nucleotide, or an inverted debasalized lock nucleic acid modification at its 5' end. For example, the sense strand contains an inverted nucleotide, an inverted debasalized nucleotide, or an inverted debasalized lock nucleic acid modification at its 5' end. For example, the sense strand of dsRNA contains an inverted nucleotide, an inverted debasalized nucleotide, or an inverted debasalized lock nucleic acid modification at its 3' end.

[0215] In one embodiment, the sense chain contains a ligand at its 3' end.

[0216] Ligand In some embodiments, long double-stranded and single-stranded oligonucleotides may include ligands. While we do not wish to be constrained by theory, ligands may modify one or more properties of the bound molecule (e.g., dsRNA described herein), including, but not limited to, pharmacodynamic, pharmacokinetic, binding, absorption, cellular distribution, cellular uptake (cellular targeting), charge, and clearance.

[0217] In one embodiment, the ligand is a targeting ligand. As used herein, “targeting ligand” means any molecule that enhances affinity to a selected target, e.g., a cell, cell type, tissue, organ, region of the body, or compartment, e.g., a cell, tissue, or organ compartment. Some exemplary targeting ligands include, but are not limited to, antibodies, antigens, folates, receptor ligands, carbohydrates, aptamers, integrin receptor ligands, chemokine receptor ligands, transferrin, biotin, serotonin receptor ligands, PSMA, endothelin, GCPII, somatostatin, LDL, and HDL ligands. Carbohydrate-based targeting ligands include, but are not limited to, D-galactose, polyvalent galactose, N-acetyl-D-galactosamine (GalNAc), polyvalent GalNAc, e.g., GalNAc2 and GalNAc3; D-mannose, polyvalent mannose, polyvalent lactose, N-acetyl-glucosamine, polyvalent fucose, glycosylated polyamino acids, and lectins. The term polyvalent indicates the presence of more than one monosaccharide unit. Such monosaccharide subunits may be linked to each other via glycosidic bonds or to scaffold molecules.

[0218] In one embodiment, the ligand is an asialoglycoprotein receptor (ASGPR) ligand. An ASGPR ligand means a ligand that binds to ASGPR. In one embodiment, the ASGPR ligand comprises one or more (e.g., one, two, three, or more) GalNAc or GalNAc derivatives linked via a divalent or trivalent branched linker. An exemplary ASGPR ligand is: [ka] Alternatively, loop-forming oligonucleotides in which 3 or 4 consecutive nucleosides are modified with a GalNAc-containing ligand; for example, [ka] (In the formula, each G* and A* is at position 2'-O) [ka] These include loop-containing oligonucleotides that have been substituted with other substances.

[0219] Exemplary carbohydrate-based targeting ligands are described in U.S. Patents 5,994,517 and 6,906,182, and PCT application PCT / US2022 / 047102, the contents of which are incorporated herein by reference in their entirety. Exemplary forate and forate analog targeting ligands are described in U.S. Patents 2,816,110; 5,552,545; 6,335,434 and 7,128,893, the contents of which are incorporated herein by reference in their entirety.

[0220] It should be noted that when two or more ligands are present, the ligands may all have the same properties, all have different properties, or some ligands may have the same properties while others have different properties. In a preferred embodiment, all ligands have different properties.

[0221] It should be noted that ligands can bind to any position on either strand of dsRNA. For example, the ligand may be at the 5' end, 3' end, or internal position of the strand, such as the sense or antisense strand of the dsRNA.

[0222] In one embodiment, the sense strand contains a ligand, i.e., the ligand is conjugated to the sense strand. For example, the ligand is conjugated to the 3' end of the sense strand.

[0223] Exemplary sense chain In one embodiment, the sense strand of dsRNA has one of the following modification patterns: [Table 3] Here, n is a 2'-O-methylnucleotide; (dN) is a 2'-deoxynucleotide; Nf is a 2'-fluoro-modified nucleotide (e.g., a 2'-deoxy-2'-fluoro-modified nucleotide); and The sense chain may include the following, as desired: (a) (i) 5'-(L1)- attached to the 5' terminal nucleotide (e.g., via the 5'-O of the terminal nucleoside) via a divalent linking group such as a phosphodiester or phosphorothioate bond, if desired; (ii) -(L2)-3' attached to the 3' terminal nucleotide (e.g., via the 3'-O of the terminal nucleoside) via a divalent linking group such as a phosphodiester or phosphorothioate bond, if desired; (iii) optionally 5'-(L1)(I)- attached to the 5' terminal nucleotide (5'-5') via a divalent linking group such as a phosphodiester or phosphorothioate bond; or (iv) -(I)(L2)-3' attached to the 3' terminal nucleotide (3'-3') via a divalent linking group such as a phosphodiester or phosphorothioate bond, if desired. A 3' or 5' terminal modification selected from the following; And here, Each (I) is an inverted nucleotide (e.g., an inverted debased nucleotide, e.g., an inverted debased ribonucleotide, e.g., an inverted debased deoxyribonucleotide); (L1) and (L2) are independently groups containing hydrogen or ligand (L), where the ligand is: (i) Lipophilic group; for example, C 10 -C 30 Alkyl, or C 10 -C 30 Alkenyl group, for example, C 10 Alkyl, C 10 Alkenil, C 12 Alkyl, C 12 Alkenil, C 14 Alkyl, C 14 Alkenil, C 15 Alkyl, C 15 Alkenil, C 16 Alkyl, C 16 Alkenil, C 18Alkyl, C 18 Alkenil, C 20 Alkyl, C 20 Alkenil, C 22 Alkyl, C 22 Alkenil, C 24 Alkyl, C 24 Contains an alkenyl group; C 15 Alkyl, C 15 Alkenil, C 17 Alkyl, C 17 Alkenil, C 19 Alkyl, C 19 Alkenil, C 21 Alkyl, C 21 Alkenil, C 23 Alkyl, or C 23 Alkenyl groups; examples include, but are not limited to, hexadecyl groups, docosanyl groups, omega-hydroxy-hexadecyl groups, and omega-hydroxy-docosanyl groups; or (ii) Receptor-targeting ligands such as groups containing ASGPR ligands; or (iii) Precursor functional groups suitable for post-synthetic functionalization with ligand groups containing or conjugating complementary reactive functional groups (e.g., as described in (i) or (ii)); Examples of precursor functional groups include amino, carboxy, primary amides (-C(O)NH2), N-succinamide, azide (-N3), mercapto (-SH), active esters (e.g., N-hydroxysuccinimide esters (NHS esters) or pentafluorophenyl esters), and 1,2,4,5-tetradinyl (e.g., 3-methyl-1,2,4,5-tetradinyl, 3-(pyridin-2-yl)-1,2,4,5-tetradinyl, or 3-(pyrimidine-2-yl)-1,2,4,5-tetradinyl); cyclooctinyl (e.g., bicyclo[6.1.0]noninyl (BCN) or dibenzocyclooctinyl (DBCO)), trans-cyclooctenyl, 2-methylsulfonylpyrimidinyl, 4-vinylpyrimidinyl, and their protected forms, including but not limited to these; or (b) Nucleotides containing the ligand modification that replaces the nucleotide; Examples of lipophilic modified nucleotides include (Nhd)-2'-O-hexadecyl-modified nucleotides; (Nda)-2'-O-docosanyl-modified nucleotides; (NhdOH)-2'-O-(omega-hydroxy-hexadecyl)-modified nucleotides; or (NdaOH)-2'-O-(omega-hydroxy-docosanyl)-modified nucleotides.

[0224] In each of the aforementioned sense strands, each nucleotide is linked sequentially (i.e., in a 3'→5' configuration) via optionally modified internucleotide bonds. For example, each nucleotide is linked by a phosphodiester or phosphorothioate internucleotide bond.

[0225] In one embodiment, counting from the 5' end of the oligonucleotide, the nucleotides at positions 1 and 2 are linked by phosphorothioate nucleotide bonds; the nucleotides at positions 2 and 3 are linked by phosphorothioate nucleotide bonds; and the remaining nucleotides are linked by phosphodiester bonds.

[0226] In one embodiment, counting from the 5' end of the oligonucleotide, the nucleotides at positions 1 and 2 are linked by phosphorothioate nucleotide bonds; the nucleotides at positions 2 and 3 are linked by phosphorothioate nucleotide bonds; the nucleotides at positions 3 and 4 are linked by phosphorothioate nucleotide bonds; and the remaining nucleotides are linked by phosphodiester bonds.

[0227] In one embodiment, when the nucleotide is m-nucleotide length, the nucleotides at positions m-1 and m, counting from the 5' end of the oligonucleotide, are linked by phosphorothioate nucleotide linkages. That is, for a nucleotide of 23 nucleotide length, the nucleotides at positions 22 and 23, counting from the 5' end of the oligonucleotide, are linked by phosphorothioate nucleotide linkages; and for a nucleotide of 21 nucleotide length, the nucleotides at positions 20 and 21, counting from the 5' end of the oligonucleotide, are linked by phosphorothioate nucleotide linkages.

[0228] In one embodiment, when the nucleotide is m-nucleotide length, the nucleotides at positions m-2 and m-1, counting from the 5' end of the oligonucleotide, are joined by phosphorothioate nucleotide bonds, and the nucleotides at positions m-1 and m are joined by phosphorothioate nucleotide bonds. That is, for a nucleotide of 23 nucleotide length, the nucleotides at positions 21 and 22, counting from the 5' end of the oligonucleotide, are joined by phosphorothioate nucleotide bonds; and positions 22 and 23 are joined by phosphorothioate nucleotide bonds; and for a nucleotide of 21 nucleotide length, the nucleotides at positions 20 and 21, counting from the 5' end of the oligonucleotide, are joined by phosphorothioate nucleotide bonds.

[0229] In another embodiment, each (inv) bonded to the 5' terminal nucleotide is linked by a phosphorothioate bond (5'-5').

[0230] In another embodiment, each (inv) bonded to the 3' terminal nucleotide is linked by a phosphorothioate bond (3'-3').

[0231] In one other embodiment, each (inv) attached to the 5' terminal nucleotide is linked by a phosphorothioate bond (5'-5'), and each (inv) attached to the 3' terminal nucleotide is linked by a phosphorothioate bond (3'-3').

[0232] In one embodiment, the sense strand follows one of S1 to S35, where one nucleotide at either position 4 to 8 or position 13 to 18 from the 5' end of the strand is replaced with a nucleotide substituted with an (L) group.

[0233] For example, in one embodiment, the sense chain of a dsNA has one of the following modification patterns:

[0234] [Table 4] [Table 5] [Table 6] Here, n is a 2'-O-methylnucleotide; (dN) is a 2'-deoxynucleotide; Nf is a 2'-fluoro-modified nucleotide; (L) is, for example, a lipophilic group (for example, C 16 or C 22 It is a ligand-modified nucleotide containing a modified (or ASGPR) ligand; and s is a phosphorothioate nucleotide bond, (s) is a phosphorothioate or phosphodiester nucleotide bond (for example, in one embodiment, each (s) is a phosphorothioate); (I) is an inverted debasalized nucleotide (e.g., an inverted debasalized ribonucleotide or an inverted debasalized deoxyribonucleotide); (L1) and (L2) are independently hydrogen- or ligand-containing groups, and the sense strand optionally contains a 5'-morpholinonucleotide, 5'-dimethylaminonucleotide, 5'-deoxynucleotide, inverted nucleotide, inverted debasalized nucleotide, or inverted debasalized lock nucleic acid modification at its 5' end.

[0235] In one embodiment, the antisense chain of the dsNA has one of the following modification patterns: [Table 7] [Table 8] Here: n is a 2'-O-methyl-modified nucleotide; s is a phosphorothioate internucleotide bond (3'-5'); (dN) is a 2'-deoxynucleotide; Nf is a 2'-fluoro-modified nucleotide; Z is [ka] And here: X is either O or S; and Each R V is independently a hydrogen or hydroxyl protecting group; and The aforementioned structure replaces the 4'-CH2OH group of the furanose (e.g., ribose) at the 5' terminal nucleotide or the 5'-CH2OH group of the pyranose ring.

[0236] In each of the aforementioned antisense strands, each nucleotide is linked sequentially (i.e., in a 3'→5' manner) via phosphodiester or phosphorothioate nucleotide-nucleotide bonds.

[0237] In any embodiment of AS1 to AS39, one nucleotide at positions 5 to 8 from the 5' end of the antisense strand is thermally destabilized (G), for example: (Ngn)-glycolic acid, S-isomer; (N2p) - 2'-phosphate nucleotide (i.e., 3'-RNA linked by 3'-5' and 2'-5' nucleotide bonds in the 5' and 3' directions, respectively); (Tna) - threose nucleotides (linked via 3'-3' and 2'-5' nucleotide bonds in the 5' and 3' directions, respectively); (MM) Nucleic acid base mismatch in sense strand; or (Nul) Unlocked nucleic acid.

[0238] In any embodiment of AS1 to AS39, nucleotide position 5, counting from the 5' end of the antisense strand, is replaced with a thermal destabilization modification (G). In any embodiment of AS1 to AS39, nucleotide position 6, counting from the 5' end of the antisense strand, is replaced with a thermal destabilization modification (G). In any embodiment of AS1 to AS39, nucleotide position 7, counting from the 5' end of the antisense strand, is replaced with a thermal destabilization modification (G). In any embodiment of AS1 to AS39, nucleotide position 8, counting from the 5' end of the antisense strand, is replaced with a thermal destabilization modification (G).

[0239] For example, the antisense chain can be selected from any of AS40 to AS58 listed in Table E: [Table 9]

[0240] In one embodiment, counting from the 5' end of the oligonucleotide, the nucleotides at positions 1 and 2 are linked by phosphorothioate nucleotide bonds; the nucleotides at positions 2 and 3 are linked by phosphorothioate nucleotide bonds; and the remaining nucleotides are linked by phosphodiester bonds. In another embodiment, counting from the 5' end of the oligonucleotide, the nucleotides at positions 1 and 2 are linked by phosphorothioate nucleotide bonds; the nucleotides at positions 2 and 3 are linked by phosphorothioate nucleotide bonds; the nucleotides at positions 3 and 4 are linked by phosphorothioate nucleotide bonds; and the remaining nucleotides are linked by phosphodiester bonds.

[0241] In one embodiment, when the nucleotide is m-nucleotide length, the nucleotides at positions m-1 and m, counting from the 5' end of the oligonucleotide, are linked by phosphorothioate nucleotide linkages. That is, for a nucleotide of 23 nucleotide length, the nucleotides at positions 22 and 23, counting from the 5' end of the oligonucleotide, are linked by phosphorothioate nucleotide linkages; and for a nucleotide of 21 nucleotide length, the nucleotides at positions 20 and 21, counting from the 5' end of the oligonucleotide, are linked by phosphorothioate nucleotide linkages.

[0242] In one embodiment, when the nucleotide is m-nucleotide length, the nucleotides at positions m-2 and m-1 are linked by phosphorothioate nucleotide linkages, and the nucleotides at positions m-1 and m, counting from the 5' end of the oligonucleotide, are linked by phosphorothioate nucleotide linkages. That is, for a nucleotide with a length of 23 nucleotides, the nucleotides at positions 21 and 22, counting from the 5' end of the oligonucleotide, are linked by phosphorothioate nucleotide linkages; and positions 22 and 23 are linked by phosphorothioate nucleotide linkages; and for a nucleotide with a length of 21 nucleotides, the nucleotides at positions 20 and 21, counting from the 5' end of the oligonucleotide, are linked by phosphorothioate nucleotide linkages.

[0243] In one embodiment, when the nucleotide is of m-nucleotide length, counting from the 5' end of the oligonucleotide: (a) The nucleotides at positions 1 and 2 are joined by a phosphorothioate nucleotide bond; (b) The nucleotides at positions 2 and 3 are linked by a phosphorothioate nucleotide bond; (c) The nucleotides at positions m-2 and m-1 are linked by phosphorothioate nucleotide bonds; and (d) The nucleotides at the m-1 and m positions are linked by phosphorothioate nucleotide bonds, The remaining nucleotides are then joined by phosphodiester bonds.

[0244] Specifically, for a nucleotide with a length of 23 nucleotides, the nucleotides at positions 1 and 2; 2 and 3; 21 and 22; and 22 and 23, counting from the 5' end of the oligonucleotide, are linked by phosphorothioate nucleotide bonds, while the remaining nucleotides are linked by phosphodiester bonds. Similarly, for a nucleotide with a length of 21 nucleotides, the nucleotides at positions 1 and 2; 2 and 3; 19 and 20; and 20 and 21, counting from the 5' end of the oligonucleotide, are linked by phosphorothioate nucleotide bonds, while the remaining nucleotides are linked by phosphodiester bonds.

[0245] In one embodiment, when the nucleotide is of m-nucleotide length, counting from the 5' end of the oligonucleotide: (a) The nucleotides at positions 1 and 2 are joined by a phosphorothioate nucleotide bond; (b) The nucleotides at positions 2 and 3 are linked by a phosphorothioate nucleotide bond; (c) The nucleotides at positions 3 and 4 are linked by a phosphorothioate nucleotide bond; and (d) The nucleotides at the m-1 and m positions are linked by phosphorothioate nucleotide bonds, The remaining nucleotides are then joined by phosphodiester bonds.

[0246] Specifically, for a nucleotide with a length of 23 nucleotides, the nucleotides at positions 1 and 2; 2 and 3; 3 and 4; and 22 and 23, counting from the 5' end of the oligonucleotide, are linked by phosphorothioate nucleotide bonds, while the remaining nucleotides are linked by phosphodiester bonds. Similarly, for a nucleotide with a length of 21 nucleotides, the nucleotides at positions 1 and 2; 2 and 3; 3 and 4; and 20 and 21, counting from the 5' end of the oligonucleotide, are linked by phosphorothioate nucleotide bonds, while the remaining nucleotides are linked by phosphodiester bonds.

[0247] For example, in one embodiment, the antisense chain of the dsNA has one of the modification patterns shown in Table F below.

[0248] [Table 10] [Table 11] [Table 12]

[0249] In any embodiment of AS58~AS135, one nucleotide at positions 5-8 from the 5' end of the antisense strand is thermally destabilized (G), for example: (Ngn)-glycolic acid, S-isomer; (N2p) - 2'-phosphate nucleotide (i.e., 3'-RNA linked by 3'-5' and 2'-5' nucleotide bonds in the 5' and 3' directions, respectively); (Tna) - threose nucleotides (linked via 3'-3' and 2'-5' nucleotide bonds in the 5' and 3' directions, respectively); (MM) Nucleic acid base mismatch in sense strand; or (Nul) Unlocked Nucleic Acid It can be replaced with.

[0250] In any embodiment of AS58 to AS135, nucleotide position 5, counting from the 5' end of the antisense strand, is replaced with a thermal destabilization modification (G). In any embodiment of AS58 to AS135, nucleotide position 6, counting from the 5' end of the antisense strand, is replaced with a thermal destabilization modification (G). In any embodiment of AS58 to AS135, nucleotide position 7, counting from the 5' end of the antisense strand, is replaced with a thermal destabilization modification (G). In any embodiment of AS58 to AS135, nucleotide position 8, counting from the 5' end of the antisense strand, is replaced with a thermal destabilization modification (G).

[0251] For example, the antisense chain can be selected from any of AS136 to AS171 listed in Table G.

[0252] [Table 13] [Table 14]

[0253] In further embodiments of each of the exemplary sense and antisense strands described herein, each of the sense strands 1-S123 can hybridize with or double-helix with any of the antisense strands AS1-AS171. In other words, the long double-stranded oligonucleotide products described herein include sense strands S1-S123 that have hybridized / double-helix-formed with antisense strands AS1-AS171. In some embodiments, the sense strand is selected from embodiments having 21 nucleotides and the antisense strand is selected from embodiments having 23 nucleotides. In some embodiments, the sense strand is selected from embodiments having 21 nucleotides and the antisense strand is selected from embodiments having 21 nucleotides. In some embodiments, the sense strand is selected from embodiments having 19 nucleotides and the antisense strand is selected from embodiments having 21 nucleotides. In some embodiments, the sense strand is selected from embodiments having 19 nucleotides and the antisense strand is selected from embodiments having 19 nucleotides.

[0254] In one embodiment, the long single-stranded oligonucleotide described herein is either one of the sense strands S1 to S123 or one of the antisense strands AS1 to AS171.

[0255] composition The dsRNAs or oligonucleotides described herein can be formulated into compositions. For example, the dsRNAs or oligonucleotides described herein can be formulated into pharmaceutical compositions for therapeutic use. In other embodiments, provided herein are pharmaceutical compositions comprising one or more dsRNAs or oligonucleotides described herein in a therapeutically effective amount, either ingested alone or formulated with one or more pharmaceutically acceptable carriers (additives), additives, and / or diluents.

[0256] Pharmaceutical compositions may be formulated in particular for administration in solid or liquid form, including those suitable for: (1) oral administration, e.g., liquid drugs (aqueous or non-aqueous or suspension), tablets, e.g., buccal, sublingual, and those targeting systemic absorption, pills, powders, granules, pastes for application to the tongue; (2) non-enteral administration, e.g., as sterile solutions or suspensions, e.g., by subcutaneous, intramuscular, intravenous or epidural injection or by sustained-release formulations; (3) topical application, e.g., as creams, ointments, or controlled-release patches or sprays applied to the skin; (4) intravaginal or rectal administration, e.g., as pessaries, creams or foams; (5) sublingual; (6) ocular; (7) transdermal; or (8) nasal. Delivery using subcutaneous or intravenous methods may be particularly advantageous.

[0257] The term "therapeutic dose" as used herein means the amount of a compound, substance, or composition containing the conjugate described herein that is effective in producing a desired therapeutic effect in at least a subpopulation of animal cells, in a reasonable benefit-to-risk ratio applicable to any medical treatment.

[0258] The term "pharmaceutically acceptable" means a compound, substance, composition, and / or dosage form that is suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications, and has a reasonable benefit / risk ratio, within the bounds of sound medical judgment.

[0259] As used herein, "pharmaceutically acceptable carriers" refer to any and all solvents, dispersions, coatings, antibacterial and antifungal agents, isotonic and absorption retarders, etc., suitable for pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. Unless any conventional media or agent is incompatible with the active compound, its use in a composition is intended. Auxiliary active compounds may also be incorporated into the composition. Pharmaceutical carriers include sterile aqueous solutions or dispersions and sterile powders for the immediate preparation of sterile injectable solutions or dispersions. The use of such media and agents with pharmaceutically active substances is well known in the art.

[0260] Pharmaceutical compositions for use in the methods described herein can be formulated in a conventional manner using one or more physiologically acceptable carriers or additives. For example, the dsRNA or oligonucleotides described herein can be formulated for administration, for example, intravenously, or orally, aerosolically, or topically. The compositions can be formulated for intrafocal, intratumoral, intraperitoneal, subcutaneous, intramuscular, or intravenous injection, infusion, liposome-mediated delivery, topical, intrathecal, gingival pocket, transrectal, intrabronchial, transnasal, transmucosal, intestinal, oral, ocular, or ear delivery.

[0261] Generally, the technology and formulations can be found in Remington's Pharmaceutical Sciences, Meade Publishing Co., Easton, PA. For systemic administration, injection is preferred, including intramuscular, intrathecal, intravenous, intraperitoneal, and subcutaneous injections. For injection, the dsRNAs described herein may be formulated in liquid solutions, preferably physiologically compatible buffers such as Hanks' solution or Ringer's solution. Furthermore, dsRNAs can be formulated in solid form and redissolved or suspended immediately before use. Lyophilized forms are also included.

[0262] dsRNA or oligonucleotides may be formulated for non-enteral administration by injection, for example, by bolus injection or continuous infusion. Injectable formulations may be provided in unit dosage forms, such as ampoules or multi-dose containers with added preservatives. Compositions may be in the form of suspensions, solutions, or emulsions in oily or aqueous media and may contain formulation aids such as suspending, stabilizing, and / or dispersing agents. Alternatively, the active ingredient may be in powder form for preparation in a suitable medium, such as sterile pyrogen-free water, before use.

[0263] LNP and liposome formulations The dsRNAs or oligonucleotides described herein may be formulated with one or more lipids for delivery. For example, the dsRNAs or oligonucleotides described herein may be formulated into lipid particles. The term “lipid particle” as used herein refers to a vesicle formed by one or more lipid components. Lipid particles are typically used as carriers for nucleic acid delivery in the context of pharmaceutical development. They act by fusing with the cell membrane and rearranging their lipid structure to deliver the drug or active pharmaceutical ingredient (API). Generally, lipid particle compositions for such delivery consist of ionizable or cationic lipids, phospholipids (particularly compounds having a phosphatidylcholine group), cholesterol, and polyethylene glycol (PEG) lipids; however, these compositions may also contain other lipids. Ionizable lipids are typically used to concentrate nucleic acid cargo at low pH and to induce membrane binding and fusing properties. Phospholipids are typically used to enhance fusing properties. Cholesterol is typically used to provide membrane integrity. PEG-lipids are typically used to provide steric stabilization. The total lipid composition typically supports surface characteristics, and therefore, the protein (opsonized) contents in biological systems thus derive their biodistribution and cellular uptake properties. Lipid particles can be lipid nanoparticles (LNPs).

[0264] In one embodiment, the lipid particles described herein include ionizable lipids. The term "ionizable lipid" as used herein refers to a lipid having at least one protonable or deprotonable group such that the lipid is positively charged at a pH below the physiological pH (e.g., pH 7.4) and neutral at a second pH, or preferably above the physiological pH. Typically, ionizable lipids are lipids containing at least one amino group that are positively charged or protonated under acidic conditions, for example, at a pH below 6.5. It will be understood by those skilled in the art that the addition or removal of protons as a function of pH is an equilibrium process, and that references to charged or neutral lipids refer to the properties of the dominant species, and that not all lipids need to be in a charged or neutral form. Generally, ionizable lipids have a pK of protonable groups in the range of about 4 to about 7. a It possesses. Ionizable lipids are also referred to here as cationic lipids.

[0265] In one embodiment, the ionizable lipid is MC3(6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl-4-(dimethylamino)butanoate (DLin-MC3-DMA or MC3). In one embodiment, the ionizable lipid is lipid ATX-002. In one embodiment, the ionizable lipid is (13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine. In one embodiment, the ionizable lipid is compound 6 or compound 22 described in WO2015 / 199952, the contents of which are incorporated herein by reference in their entirety.

[0266] While not limited to ionizable lipids, ionizable lipids can constitute 20-90% (mol) of the total lipids present in lipid particles.

[0267] The term "noncationic lipid" as used herein refers to all amphiphilic lipids as well as all other neutral or anionic lipids. Therefore, noncationic lipids can be neutral, uncharged, zwitterionic, or anionic lipids. Noncationic lipids are typically used to enhance fusion properties.Exemplary noncationic lipids include distearoyl-sn-glycerol-phosphoethanolamine, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), and palmitoyloleoylphosphatidylethanolamine. POPE, dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), monomethyl-phosphatidylethanolamine (e.g., 16-O-monomethylPE), dimethyl-phosphatidylethanolamine (e.g., 16-O-dimethylPE), 18-1-trans PE, 1-Stearoyl-2-Oleoyl-Phosphatidiethanolamine (SOPE), Hydrogenated Soybean Phosphatidylcholine (HSPC), Egg Phosphatidylcholine (EPC), Dioleoyl Phosphatidylserine (DOPS), Sphingomyelin (SM), Dimyristoyl Phosphatidylcholine (DMPC), Dimyristoyl Phosphatidylglycerol (DMPG), Distearoyl Phosphatidylglycerol (DSPG), Diylcoyl Phosphatidylcholine (DEPC), Palmitoyl Oleoyl Phosphatidyl This includes, but is not limited to, lyserol (POPG), dierydoyl-phosphatidylethanolamine (DEPE), lecithin, phosphatidylethanolamine, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, egg sphingomyelin (ESM), cephalin, cardiolipin, phosphatidic acid, cerebroside, dicetylphosphate, lysophosphatidylcholine, dilinoleoylphosphatidylcholine, or mixtures thereof.It is understood that other diacylphosphatidylcholine and diacylphosphatidylethanolamine phospholipids can also be used. The acyl group of these lipids is preferably C. 10 -C 24 The acyl group is derived from a fatty acid having a carbon chain, such as lauroyl, myristoyl, palmitoyl, stearoyl, or oleoyl.

[0268] Other examples of noncationic lipids suitable for use in lipid particles include nonphospholipids such as stearylamine, dodecylamine, hexadecylamine, acetyl palmitate, glycerol lysine oleate, hexadecyl telate, isopropyl myristate, amphoteric acrylic acid polymers, triethanolamine-lauryl sulfate, alkyl-aryl sulfate polyethyloxylated fatty acid amides, dioctadecyldimethylammonium bromide, ceramide, and sphingomyelin.

[0269] In one embodiment, the noncationic lipid is a phospholipid. In one embodiment, the noncationic lipid is selected from DSPC, DPPC, DMPC, DOPC, POPC, DOPE, and SM. In one preferred embodiment, the noncationic lipid is DPSC.

[0270] Noncationic lipids can constitute 0-30% (mol) of the total lipids present in the lipid particles. For example, the noncationic lipid content is 5-20% (mol) or 10-15% (mol) of the total lipids present in the lipid particles. In various embodiments, the molar ratio of ionizable lipids to neutral lipids is in the range of approximately 2:1 to approximately 8:1.

[0271] In some embodiments, lipid particles may further contain conjugated lipid molecules. As used herein, “conjugated lipid” refers to a lipid molecule conjugated with a non-lipid molecule such as PEG, polyoxazoline, polyamide, or polymer (e.g., cationic polymer). Generally, these lipids are used to prevent aggregation of lipid particles and / or provide steric stabilization. Exemplary conjugated lipids include, but are not limited to, PEG-lipid conjugates, polyoxazoline (POZ)-lipid conjugates, polyamide-lipid conjugates (e.g., ATTA-lipid conjugates), cationic-polymer lipid (CPL) conjugates, and mixtures thereof. In some embodiments, the conjugated lipid molecule is a PEG-lipid conjugate, e.g., a (methoxypolyethylene glycol)-conjugated lipid.

[0272] Exemplary PEG-lipid conjugates include, but are not limited to, PEG-diacylglycerol (DAG) (e.g., l-(monomethoxy-polyethylene glycol)-2,3-dimyristoylglycerol (PEG-DMG)), PEG-dialkyloxypropyl (DAA), PEG-phospholipids, PEG-ceramide (Cer), pegylated phosphatidylethanolamine (PEG-PE), PEG succinate diacylglycerol (PEGS-DAG) (e.g., 4-O-(2',3'-di(tetradecanoyloxy)propyl-1-O-(w-methoxy(polyethoxy)ethyl)butanediolate (PEG-S-DMG)), PEG-dialkoxypropylcarbam, N-(carbonyl-methoxypolyethylene glycol 2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine sodium salt, or mixtures thereof.

[0273] PEG-DAA conjugates may be, for example, PEG-dilauryloxypropyl, PEG-dimyristyloxypropyl, PEG-dipalmityloxypropyl, or PEG-distearyloxypropyl. PEG-lipids include PEG-DMG, PEG-dilaurylglycerol, PEG-dipalmitoylglycerol, PEG-disterylglycerol, PEG-dilaurylglycamide, PEG-dimyristylglycamide, PEG-dipalmitoylglycamide, PEG-disterylglycamide, PEG-cholesterol (1-[8'-(cholest-5-ene-3[beta]-oxy)carboxamide-3',6'-dioxaoctanyl]carbamoyl-[omega]-methyl-poly(ethyleneglycol) The lipids may be PEG-DMB (3,4-ditetradecoxylbenzyl-[omega]-methyl-poly(ethylene glycol) ether) and 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000]. As some examples, the PEG-lipid may be one or more of PEG-DMG, 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000].

[0274] PEG or conjugated lipids can constitute 0-20% (mol) of the total lipids present in the lipid particles. In one embodiment, the PEG or conjugated lipid content is 0.5-10% or 2-5% (mol) of the total lipids present in the lipid particles.

[0275] In one embodiment, the lipid particles may further contain components such as sterols to provide membrane integrity. Exemplary sterols that can be used in lipid particles are cholesterol and its derivatives. Non-limiting examples of cholesterol derivatives include polar analogs such as 5α-cholestanol, 5β-coprostanol, cholesteryl-(2'-hydroxy)-ethyl ether, cholesteryl-(4'-hydroxy)-butyl ether, and 6-ketocholestanol; non-polar analogs such as 5α-cholestanol, cholestenone, 5α-cholestanone, 5β-cholestanone, and cholesteryl decanoate; and mixtures thereof. In one embodiment, the cholesterol derivative is a polar analog such as cholesteryl-(4'-hydroxy)-butyl ether.

[0276] Components that provide membrane integrity, such as sterols, can constitute 0-50% (mol) of the total lipids present in the lipid particles. In one embodiment, such components constitute 20-50% (mol) or 30-40% (mol) of the total lipid content of the lipid particles.

[0277] In one embodiment, the lipid particles include ionizable lipids; noncationic lipids; conjugated lipids that prevent particle aggregation; and sterols. The molar ratios of ionizable lipids, noncationic lipids, sterols, and PEG / conjugated lipids may vary as needed. For example, the lipid particles may contain 30-70% ionizable lipids by moles or total weight of the composition, 0-60% cholesterol by moles or total weight of the composition, 0-30% noncationic lipids by moles or total weight of the composition, and 1-10% conjugated lipids by moles or total weight of the composition. Preferably, the composition contains 30-40% ionizable lipids by moles or total weight of the composition, 40-50% cholesterol by moles or total weight of the composition, and 10-20% noncationic lipids by moles or total weight of the composition. In another embodiment, the composition comprises 50-75% ionizable lipids by mole or total weight, 20-40% cholesterol by mole or total weight, 5-10% noncationic lipids by mole or total weight, and 1-10% conjugated lipids by mole or total weight. The composition may also contain 60-70% ionizable lipids by mole or total weight, 25-35% cholesterol by mole or total weight, and 5-10% noncationic lipids by mole or total weight. The composition may also contain up to 90% ionizable lipids and 2-15% noncationic lipids by mole or total weight.The formulation may, for example, contain 8-30% ionizable lipids by mole or total weight of the composition, 5-30% noncationic lipids by mole or total weight of the composition, and 0-20% cholesterol by mole or total weight of the composition; 4-25% ionizable lipids by mole or total weight of the composition, 4-25% noncationic lipids by mole or total weight of the composition, 2-25% cholesterol by mole or total weight of the composition, 10-35% conjugate lipids by mole or total weight of the composition, and 5% cholesterol by mole or total weight of the composition; or the composition may contain Lipid particle formulations may also contain 2-30% ionizable lipids by mole or total weight, 2-30% noncationic lipids by mole or total weight of composition, 1-15% cholesterol by mole or total weight of composition, 2-35% conjugate lipids by mole or total weight of composition, and 1-20% cholesterol by mole or total weight of composition; or up to 90% ionizable lipids by mole or total weight of composition and 2-10% noncationic lipids by mole or total weight of composition, or 100% cationic lipids by mole or total weight of composition. In one embodiment, the lipid particle formulation contains ionizable lipids, phospholipids, cholesterol and PEG-ylated lipids in a molar ratio of 50:10:38.5:1.5. In another embodiment, the lipid particle formulation contains ionizable lipids, cholesterol and PEG-ylated lipids in a molar ratio of 60:38.5:1.5.

[0278] In one embodiment, the lipid particles contain approximately 20 mol% to 90 mol% of ionizable lipids of the total lipids present in the particles; approximately 5 mol% to 30 mol% of noncationic lipids of the total lipids present in the particles; approximately 0.5 mol% to 20 mol% of conjugated lipids that prevent particle aggregation of the total lipids present in the particles; and approximately 20 mol% to 50 mol% of sterols of the total lipids present in the particles. In one embodiment, the lipid particles contain ionizable lipids / noncationic lipids / sterols / conjugated lipids in a molar ratio of 50:10:38.5:1.5.

[0279] In one embodiment, the total lipid to diffusion (mass or weight) ratio is about 10:1 to about 30:1. The amounts of lipids and nucleic acids can be adjusted to provide a desired N / P ratio, for example, 3, 4, 5, 6, 7, 8, 9, 10 or higher.

[0280] Administration to the target The dsRNAs and / or oligonucleotides described herein, like other pharmaceuticals, can be formulated for administration in any convenient manner for use in pharmaceuticals, such as human or animal pharmaceuticals.

[0281] The dsRNA and / or oligonucleotides described herein, or pharmaceutical compositions containing them, can be administered to subjects using a variety of delivery routes. Exemplary delivery routes include, but are not limited to, intravenous, subcutaneous, topical, rectal, anal, vaginal, nasal, lung, and ocular.

[0282] The dsRNAs and / or oligonucleotides described herein can be administered in several ways, depending on whether topical or systemic treatment is desired and the treatment area. Administration may be non-enteral, topical (including ocular, vaginal, rectal, nasal, and percutaneous), or oral. Non-enteral administration includes intravenous infusion, subcutaneous, intraperitoneal, or intramuscular injection, or intrathecal or intraventricular administration.

[0283] The route and site of administration may be selected to enhance targeting. For example, intramuscular injection into the target muscle is a logical choice for targeting muscle cells. Lung cells are targeted by administration of the dsRNA and / or oligonucleotides described herein in aerosol form. Vascular endothelial cells are targeted by coating a balloon catheter with the dsRNA and / or oligonucleotides described herein and mechanically introducing the dsRNA and / or oligonucleotides described herein.

[0284] In one embodiment, the present invention provides a method for administering the dsRNA and / or oligonucleotides described herein to a subject (e.g., a human subject). In another embodiment, the present invention relates to the dsRNA and / or oligonucleotides described herein for use in inhibiting the expression of a target gene in a subject. The method or medical use involves administering a unit dose of the dsRNA and / or oligonucleotides described herein.

[0285] The defined dose may be an effective amount for treating or preventing a disease or disorder, such as a disease or disorder associated with the target gene. For example, a unit dose may be administered by injection (e.g., intravenous, subcutaneous, or intramuscular), inhalation, or topical application.

[0286] In one embodiment, the unit dose is administered at a frequency of once a day or less, for example, every two days, every four days, every eight days, or every 30 days or less. In another embodiment, the unit dose is not administered at any frequency (e.g., not regularly). For example, the unit dose may be administered as a single dose.

[0287] In one embodiment, the effective dose is administered together with other traditional therapeutic modalities.

[0288] The dsRNAs and / or oligonucleotides described herein are administered in several ways to mammals, particularly non-human primates or large mammals such as humans.

[0289] In one embodiment, administration of the dsRNA and / or oligonucleotide compositions described herein may be non-enteral, e.g., intravenous (e.g., bolus or diffuse infusion), intradermal, intraperitoneal, intramuscular, intrathecal, intraventricular, intracranial, subcutaneous, transmucosal, buccal, sublingual, endoscopic, rectal, oral, vaginal, topical, pulmonary, intranasal, urethral, ​​or ocular. Administration may be performed by the subject or by another person, e.g., a healthcare professional. The medication may be provided in a quantitative or quantitatively delivering dispenser.

[0290] In one embodiment, the dsRNA and / or oligonucleotides described herein are administered subcutaneously or intravenously.

[0291] In one embodiment, the dsRNA and / or oligonucleotides described herein are administered intrathecally.

[0292] In one embodiment, the dsRNA and / or oligonucleotides described herein are administered by intravitreous administration.

[0293] cell The present invention also provides cells comprising the compounds, dsRNA, or oligonucleotides described herein. The term “cell” as used herein refers to a single cell and a population of cells (i.e., more than one). A cell may be a prokaryotic or eukaryotic cell. Exemplary cells include, but are not limited to, bacterial cells, yeast cells, plant cells, animal (including insect) or human cells. In some embodiments, a cell is a eukaryotic cell. For example, a cell is a mammalian cell. It should be noted that a cell may be in vivo, in vitro, or ex vivo.

[0294] kit The dsRNA or oligonucleotide described herein may be provided in a kit, for example, as a component of the kit. For example, the kit includes (a) the dsRNA or oligonucleotide described herein, and optionally (b) informational material. The informational material may be descriptive, instructive, sales, or other material relating to the method described herein and / or the dsRNA or oligonucleotide described herein for the method described herein. The informational material of the kit is not limited in form. In some embodiments, the informational material may include information about the product, such as the dsRNA or oligonucleotide, its molecular weight, concentration, expiration date, batch, or production site information. In some embodiments, the informational material relates to the use of the dsRNA or oligonucleotide for the treatment, prevention, or diagnosis of disorders and conditions.

[0295] In one embodiment, the information material may include, for example, instructions for administering dsRNA or oligonucleotides in an appropriate form for carrying out the method described herein in an appropriate dose, dosage form, or method of administration (e.g., the dose, dosage form, or method of administration described herein). In another embodiment, the information material may include instructions for administering dsRNA or oligonucleotides to an appropriate subject, for example, a human being, for example, a human being who has or is at risk of having a disorder or condition requiring treatment.

[0296] The information materials included in the kit are not limited in form. While information materials, such as instructions, are often provided in writing, they may also be in other forms, such as computer-readable materials.

[0297] The components of the kit, such as dsRNA or oligonucleotides, may be supplied in any form, e.g., liquid, dry, or lyophilized. It is preferable that the dsRNA or oligonucleotides are substantially pure and / or sterile. When the dsRNA or oligonucleotides are supplied in a liquid solution, the liquid solution is preferably an aqueous solution, and a sterile aqueous solution is preferred. When the dsRNA or oligonucleotides are supplied in dry form, reconstitution is generally performed by adding a suitable solvent. A solvent, such as sterile water or a buffer, may be provided in the kit if desired.

[0298] A kit may include one or more containers for the components of the kit. In one embodiment, a kit may include separate containers, dividers, or compartments for different components of the kit. For example, dsRNA or oligonucleotides may be contained in a bottle, vial, or syringe, and informational material may accompany the container. In another embodiment, separate elements of the kit are contained in a single, undivided container. For example, dsRNA or oligonucleotides are contained in a bottle, vial, or syringe with informational material in the form of a label attached. In one embodiment, a kit may include multiple (e.g., packs) of individual containers, each containing one or more units of dsRNA or oligonucleotide dosage forms. For example, a kit may include multiple syringes, ampoules, foil packets, or blister packs, each containing a single-unit dose of dsRNA or oligonucleotide. The containers of the kit may be airtight, waterproof (e.g., impermeable to changes in moisture or evaporation), and / or light-shielded.

[0299] The kit may optionally include a device suitable for administering dsRNA or oligonucleotides, such as a syringe, inhalant, dropper (e.g., eye drops), swab (e.g., cotton swab or wooden swab), or any such delivery device. In one embodiment, the device is an implantable device for dispensing a quantitative amount of dsRNA or oligonucleotide. The present invention also relates to a method of providing a kit, for example, by combination of the components described herein.

[0300] In one embodiment, the kit may further include additional components and / or reagents for carrying out the method described herein using the dsRNA or oligonucleotide described herein.

[0301] Methods for inhibiting the expression of target genes Aspects of the present invention also relate to a method for inhibiting the expression of a target gene in a subject. The method comprises administering to a subject in an amount sufficient to inhibit the expression of the target gene, (i) a double-stranded RNA described herein (where the antisense strand is substantially complementary to the target gene); and / or (ii) an oligonucleotide described herein (where the oligonucleotide is substantially complementary to the target gene).

[0302] The present invention further relates to the use of dsRNA and / or oligonucleotides for inhibiting the expression of target genes in target cells. The present invention further relates to the use of the oligonucleotides and / or dsRNA molecules described herein for inhibiting the expression of target genes in target cells in vitro.

[0303] In other embodiments, the present invention provides a method for modulating the expression of a target gene in cells, comprising administering the cells a dsRNA and / or oligonucleotide described herein. It should be noted that administration to cells may be in vitro or in vivo. Methods for administering compounds to cells are well known and available to those skilled in the art. Administration of the compounds used herein to cells involves contacting the cells with the compound so that the compound is taken up by the cells. Generally, cells can be contacted with dsRNA / oligonucleotides in cell culture, for example, in vitro or ex vivo, or compounds can be administered to a subject, for example, in vivo. The terms “contact” or “contact” as used herein in relation to cell contact include immersing cells in a suitable culture medium containing the dsRNA and / or oligonucleotide described herein. When cells are in vivo, “contact” or “contact” includes, for example, administering the dsRNA and / or oligonucleotide described herein in a pharmaceutical composition to a subject via a suitable administration route so that the compound contacts the cells in vivo. For example, when the cells are in vitro, administration to the cells includes placing the cells in a suitable culture medium containing dsRNA / oligonucleotide. When the cells are in vivo, administration to the cells includes targeting the sRNA / oligonucleotide via a suitable administration route to ensure contact with the cells in vivo.

[0304] In one embodiment, the target genes are factor VII, Eg5, PCSK9, TPX2, apoB, SAA, TTR, RSV, PDGF beta gene, Erb-B gene, Src gene, CRK gene, GRB2 gene, RAS gene, MEKK gene, JNK gene, RAF gene, Erk1 / 2 gene, PCNA(p21) gene, MYB gene, JUN gene, FOS gene, BCL-2 gene, hepcidin, activated protein C, cyclin D gene, VEGF gene, EGFR gene, and cy The mutations are selected from the group consisting of mutations in the following genes: Clin A gene, cyclin gene, WNT-1 gene, beta-catenin gene, c-MET gene, PKC gene, NFKB gene, STAT3 gene, survivor gene, Her2 / Neu gene, topoisomerase I gene, topoisomerase II alpha gene, p73 gene, p21 (WAF1 / CIP1) gene, p27 (KIP1) gene, PPM1D gene, RAS gene, caveolin I gene, MIB I gene, MTAI gene, M68 gene, tumor suppressor gene, and p53 tumor suppressor gene.

[0305] In another embodiment, the target gene is selected from the group consisting of APP, ATXN2, C9orf72, TARDBP, MAPT(Tau), HTT, SNCA, FUS, ATXN3, ATXN1, SCA1, SCA7, SCA8, MeCP2, PRNP, SOD1, DMPK, TTR, SCN9A, LRRK2, GPR75, APOE, SCD5, ELOVL1, FLNA, ALK, CHI3L1(YKL-40), RPS25, α2-AR, and GSK3α.

[0306] In one embodiment, the target genes are myostatin (MSTN); choline receptor nicotin alpha-1 subunit (CHRNA1); choline receptor nicotin beta-1 subunit (CHRNB1); choline receptor nicotin delta subunit (CHRND); choline receptor nicotin epsilon subunit (CHRNE); choline receptor nicotin gamma subunit (CHRNG); collagen type XIII alpha-1 chain (COL13A1); docking protein 7 (DOK7); LDL receptor-related protein 4 (LRP4); muscle-related receptor tyrosine kinase (MUSK); synaptic protein-associated receptor (RAPSN); voltage-gated sodium channel alpha subunit 4 (SCN4A); and double homeobox 4 (DUX4), myotonic dystrophy protein kinase (DMPK), glycogen synthase 1 (GYS1), motor neuron survival 1 (SMN1), and alpha-glucosidase (GAA). ;Selected from the group consisting of: adrenaline receptor beta-1 (ADRB1); voltage-gated calcium channel subunit alpha-1C (CACNA1C); voltage-gated calcium channel subunit alpha-1G (CACNA1G) (T-type calcium c channel); angiotensin II receptor type 1 (AGTR1); voltage-gated sodium channel alpha subunit 2 (SCN2A); hyperpolarization-activated cyclic nucleotide-dependent potassium channel 1 (HCN1); hyperpolarization-activated cyclic nucleotide-dependent potassium channel 4 (HCN4); hyperpolarization-activated cyclic nucleotide-dependent potassium channel 3 (HCN3); potassium voltage-gated channel subfamily A member 5 (KCNA5); inward-rectifying potassium channel subfamily J member 3 (KCNJ3); inward-rectifying potassium channel subfamily J member 4 (KCNJ4); phospholamban (PLN); calcium / calmodulin-dependent protein kinase II delta (CAMK2D); or phosphodiesterase 1 (PDE1).

[0307] In one embodiment, the target gene is selected from the group consisting of myostatin (MSTN); choline receptor nicotin alpha-1 subunit (CHRNA1); choline receptor nicotin beta-1 subunit (CHRNB1); choline receptor nicotin delta subunit (CHRND); choline receptor nicotin epsilon subunit (CHRNE); choline receptor nicotin gamma subunit (CHRNG); collagen type XIII alpha-1 chain (COL13A1); docking protein 7 (DOK7); LDL receptor-related protein 4 (LRP4); muscle-related receptor tyrosine kinase (MUSK); synaptic protein-associated receptor (RAPSN); voltage-gated sodium channel alpha subunit 4 (SCN4A); and double homeobox 4 (DUX4), myotonic dystrophy protein kinase (DMPK), glycogen synthase 1 (GYS1), motor neuron survival 1 (SMN1), and alpha-glucosidase (GAA).

[0308] In one embodiment, the target gene is selected from the group consisting of adrenaline receptor beta-1 (ADRB1); voltage-gated calcium channel subunit alpha-1C (CACNA1C); voltage-gated calcium channel subunit alpha-1G (CACNA1G) (T-type calcium c channel); angiotensin II receptor type 1 (AGTR1); voltage-gated sodium channel alpha subunit 2 (SCN2A); hyperpolarization-activated cyclic nucleotide-dependent potassium channel 1 (HCN1); hyperpolarization-activated cyclic nucleotide-dependent potassium channel 4 (HCN4); hyperpolarization-activated cyclic nucleotide-dependent potassium channel 3 (HCN3); potassium voltage-gated channel subfamily A member 5 (KCNA5); inward-rectifying potassium channel subfamily J member 3 (KCNJ3); inward-rectifying potassium channel subfamily J member 4 (KCNJ4); phospholamban (PLN); calcium / calmodulin-dependent protein kinase II delta (CAMK2D); or phosphodiesterase 1 (PDE1).

[0309] In one embodiment, the target gene is selected from the group consisting of MUC5B, TSLP, IL33, ALOX15, AGER (RAGE), MUC5AC, and STAT6.

[0310] In one embodiment, the target gene is selected from the group consisting of delta-4-desaturase, sphingolipid 1 (DEGS1); leptin; horiclin (FLCN); zinc finger protein 423 (ZFP423); cyclin-dependent kinase 6 (CDK6); regulatory protein 1 of the MTOR complex (RPTOR); mechanical target of rapamycin kinase (mTOR); forkhead box P1 (FOXP1); phosphodiesterase 3B (PDE3B); and activin A receptor type 1C (ACVR1C).

[0311] In one embodiment, the target gene is selected from the group consisting of TTR (CNS, ocular, and systemic) for hATTR, myocilin (MYOC), Ras homolog family member A (RhoA), SSB (small RNA-binding exonuclease protective factor La), optinulin, carbonic anhydrase 2 (CA2), Rho-related coiled-coil protein kinase 1 (ROCK1), Rho-related coiled-coil protein kinase 2 (ROCK2), angiopoietin-like 7 (ANGPTL7), and cytochrome P450 1B1 (CYP1B1).

[0312] In other embodiments, the target genes are PPARG, ADIPOQ, CD36, LPL, ADAMTS9, RASD1, GYS2, CAT, DPYS, MLXIPL, VEGFA, HLA-DQA1, LIPA, CTSC, FCGR2A, GBE1, SH2B3, CTSK, CDKN2B, ELN, ARG1, HHEX, TCF7L2, CYP2A6, ALDH2, ACADS, GLYCTK, LDLR, HAL, ACER3, SLC7A7, PTPN22, CDKN1C, LEPR, SNAI2, PGM1, IGF2BP2, TTPA, ATP7B, ASPA, ADRB3, MAN2B1, RCAN1, PIGL, TBX1, LMNB1, FBP1, ETFA, LMNA, LAT2, PRKAG2, SELENBP1, TKT, PCSK1, PSAP, NDN, ACY1, SATB2, CYP21A2, POMC, CDC73, CTSH, CFTR, CTSA, G6PD, EXT1, EXT2, CPT1A, SEMA5A, WFS1, Kit, ACAT1, GGCX, FKBP6, PPARGC1B, DGCR6, HMGCS2, PEPD, WRN, LCAT, KLF13, SLC16A2, DHCR7, ITPR3, CLDN4, FZD9, SLC30A2, APOA, HADHA, CDKAL1, PTPN2, LIPC, CD226, PON1, MCCC1, EIF2AK3, GYG1, BCL7B, AGL, VKORC1, BAZ1B, NAGS, ASL, STAR, ACP2, POLG, GAA, ALDH3A2, MANBA, ARSA, AGA, CYP27B1, CPS1, DLAT, DCXR, EIF4H, DYRK1A, GTF2I, LAMP, CTH, EPO, FLAD1, AKT2, WAC, GLB1, RFC2, BACH2, D2HGDH, GHRL, TBL2, RRM2B, PRKACA, DLD, NEU1, ADSL, SLC22A5, ADCY10, INSR, HSD17B10, DGCR8, NPAP1, OXCT1, SDC3, HMGCL, PGAP1, MCCC2, LMF1, PIGM, UCP3, PAH, VPS33A, BCS1L, PDP1, AHCY, ALDH18A1, ENO3, MTTP, MAT1A, GNPTAB, PHGDH, BCAT2, CBS, HDAC4, LIG3, PSAT1, HGD, CTNND2, PDHB, PDHA1, NADK2, UPB1PKLR, BCKDK, MEN1, GALT, LIMK1, SLC39A4, KCNJ11, PDHX, ACAD8, GSS, CHRNA7, SLC6A9, ERBB3, GLUD1, GSR, OAT, SLC6 A8, CLIP2, STX1A, CARTPT, SLC25A15, DGCR2, LIPT2, NR5A1, DNM1L, PHEX, SLC30A9, B3GAT3, SLC34A3, SLC12A3, EPX, S The group is selected from ARS2, CAPN10, ASNS, ALDOB, AGRP, MFF, GK, APOC2, CLDN3, HPRT1, PFKM, AMACR, SNRPN, HNF1B, L2HGDH, SORD, IDH2, TPMT, CYP2C19, TERT, MC4R, TMPRSS15, SLCO1B3, FGF23, PAX4, SLC30A8, MTNR1B, SI, SLCO1B1, and NR0B2.

[0313] Reactive phosphorus group The various embodiments described herein include reactive phosphorus groups. While we do not wish to be bound by theory, reactive phosphorus groups are useful in the formation of nucleoside bonds, including, for example, between phosphodiesters and phosphorothioate nucleoside bonds. Such reactive phosphorus groups are known in the art and include, but are not limited to, phosphoramidites, H-phosphonates, alkyl-phosphonates, phosphate triesters, and phosphorus-containing chiral additives. III or P V Contains phosphorus atoms in the valence state. Reactive phosphites include phosphoramidites (P III A reactive phosphorus group in the chemical form is a preferred reactive phosphorus group for solid-phase oligonucleotide synthesis. The intermediate phosphite compound is then oxidized to the Pv state using a known method to obtain a phosphodiester or phosphorothioate nucleoside bond.

[0314] In one embodiment, the reactive phosphorus group is -P(OR P1 )N(R P2 )2, -P(SR P1 )N(R P2 )2, -P(O)(OR P1 )N(R P2)2, -P(S)(OR P1 )N(R P2 )2, -P(R P3 )N(R P2 )2, -P(O)(SR P1 )N(R P2 )2, -P(O)(OR P1 )H, -P(S)(OR P1 )H, -P(O)(SR P1 )H, -P(O)(OR P1 )R P3 , -P(S)(OR P1 )R P3 , or -P(O)(SR P1 )R P3 That is the case.

[0315] One reason, R P1 C is replaced as desired. 1-6 It is alkyl. For example, R P1 Ha is, as desired, OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, NH(C1-C4)alkyl, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8)alkyl, O(C1-C8)alkyl (i.e., C1- C8 alkoxy), O(C1-C8) haloalkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) p C is substituted with one, two, three, four, or five substituents independently selected from the group consisting of -NH2 or CH2-aryl-alkoxy. 1-6It is an alkyl group, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6. In one embodiment, R P1 C is replaced with CN or -SC(O)Ph as desired. 1-6 It is alkyl. For example, R P1 It is cyanoethyl (-CH2CH2CN).

[0316] One way of doing this is for each R P2 C is independently replaced as desired. 1-6 It is alkyl. For example, each R P2 R can be independently selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl. Two or more R P2 It should be noted that when the groups are present on the reactive phosphorus group, they may be the same or different. Therefore, in a certain non-restrictive example, two or more R groups P2 When a base exists, R P2 The basis is different. In some other non-restrictive example, two or more R P2 When a base exists, R P2 The basis is the same. In one embodiment, each R P2 It is isopropyl.

[0317] One Tuesday, both R P2These form 3- to 8-membered heterocyclines, which are optionally substituted with the nitrogen atom to which they are bonded. Exemplary heterocyclines include, but are not limited to, pyrrolidinyl, piperazinyl, dioxanyl, morpholinyl, tetrahydrofuranyl, piperidyl, 4-morpholyl, 4-piperazinyl, pyrrolidinyl, perhydropyrrolidinyl, 1,4-diazaperhydroepinyl, 1,3-dioxanyl, and 1,4-dioxanyl, and each of these may optionally contain OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, and NH(C1-C4)alkyl. L, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8)alkyl, O(C1-C8)alkyl (i.e., C1-C8 alkoxy), O(C1-C8)haloalkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) p It may be substituted with one, two, or three substituents independently selected from -NH2 or CH2-aryl-alkoxy, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6.

[0318] One reason, R P1 and R P2One of these atoms combines with the atom to which they are bonded to form a 4- to 8-membered heterocycline, which is optionally substituted. Exemplary heterocyclines include, but are not limited to, pyrrolidinyl, piperazinyl, dioxanyl, morpholinyl, tetrahydrofuranyl, piperidyl, 4-morpholyl, 4-piperazinyl, pyrrolidinyl, perhydropyrrolidinyl, 1,4-diazaperhydroepinyl, 1,3-dioxanyl, and 1,4-dioxanyl, and each of these may optionally contain OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, and NH(C1-C4)alkyl. L, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8)alkyl, O(C1-C8)alkyl (i.e., C1-C8 alkoxy), O(C1-C8)haloalkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) p It may be substituted with one, two, or three substituents independently selected from -NH2 or CH2-aryl-alkoxy, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6.

[0319] In the reactive phosphorus group, each R P3 C is independently replaced as desired. 1-30 Alkyl-substituted C1-C 30 Alkyl, optionally substituted C2-C 30 Alkenyl, or optionally substituted C2-C 30 Alkinyl (for example, C1-C which is substituted as desired) 10 Alkyl, optionally substituted C2-C10 Alkenyl, or optionally substituted C2-C 10 It is alkinyl). For example, R P3 The following are available upon request: OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, NH(C1-C4)alkyl, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8)alkyl, O(C1-C8)alkyl (i.e., C1-C 8 alkoxy), O(C1-C8) haloalkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) p C is substituted with one, two, three, four, or five substituents independently selected from the group consisting of -NH2 or CH2-aryl-alkoxy. 1-6 It may be an alkyl group, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6. For example, R P3 The compound is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, pentyl, or hexyl, each of which may be substituted as desired with NH2, OH, C(O)NH2, COOH, halo, SH, or C1-C6 alkoxy.

[0320] In one embodiment, the reactive phosphorus group is -P(OR P1 )(N(R P2 )2) For example, the reactive phosphorus group is -P(OR P1 )(N(R P2 )2) and here, R P1 It is 2-cyanoethyl (-CH2CH2CN), and each R P2 It is isopropyl.

[0321] Some exemplary aspects of the present invention are described by one or more of the following numbered embodiments:

[0322] Embodiment 1: Formula (A): [ka] (Formula A) [During the ceremony: B is a nucleic acid base that is modified as desired; X S is O, CH2, S, or NH; R 22 C is substituted with hydroxyl, protected hydroxyl, halogen, or optionally. 1-30 Alkoxy (e.g., methoxy, 2-methoxyethoxy), alkoxyalkyl (e.g., 2-methoxyethyl), hydrogen, optionally substituted C 1-30 Alkyl, optionally substituted C 2-30 Alkenyl, optionally substituted with C 2-30 Alkynyl, alkoxyalkylamine, alkoxyoxycarboxylate, amino, alkylamino, dialkylamino, 5-8 membered heterocyclyl, -OC 4-30 Alkyl-ON(CH2R 8 )(CH2R 9 ), -OC 4-30 Alkyl-ON(CH2R 8 )(CH2R 9 ), ligand, linker covalently bound to one or more ligands, or subsequent binding to internucleotide bonds of nucleosides; R 23 The subsequent bonding of nucleotides to the nucleoside, hydroxyl, protective hydroxyl, halogen, or optionally substituted C 2-30 Alkinyl, optionally substituted with C 1-30 Alkoxy (e.g., methoxy, 2-methoxyethoxy), alkoxyalkyl (e.g., 2-methoxyethyl), hydrogen, optionally substituted C 1-30 Alkyl, optionally substituted C2-30 Alkenyl, alkoxyalkylamine, alkoxyoxycarboxylate, amino, alkylamino, dialkylamino, 5-8 membered heterocyclyl, -OC 4-30 Alkyl-ON(CH2R 8 )(CH2R 9 ), -OC 4-30 Alkyl-ON(CH2R 8 )(CH2R 9 ), a phosphate group, a ligand, or a linker covalently bonded to one or more ligands; R 24 C is substituted with hydrogen, if desired. 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted with C 2-6 Alkinyl, or C as desired. 1-6 It is an alkoxy; or R 22 and R 24 They become one 4'-C(R 10 R 11 ) v -Y-2' or 4'-YC(R 10 R 11 ) v -2' is; Y is -O-, -CH2-, -CH(Me)-, -C(CH3)2-, -S-, -N(R 12 )-, -C(O)-, -C(S)-, -S(O)-, -S(O)2-, -OC(O)-, -C(O)O-, -N(R 12 )C(O)-, or -C(O)N(R 12 )-and; R 10 and R 11 These are independently H, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, or optionally substituted C2-C6 alkynyl; R 12 C is substituted with hydrogen, if desired. 1-30 Alkyl, optionally substituted C1-C 30 Alkoxy, C 1-4 Haloalkyl, optionally substituted C2-4 Alkenyl, optionally substituted with C 2-4 Alkinyl, optionally substituted with C 1-30 It is an alkyl-CO2H or nitrogen-protecting group; v is 1, 2, or 3; and R 5 ha-L 5 -CH=CH-X P And, Here, L 5 is a combination or C 1-30 It is an alkylene (for example, L 5 is a combination, that is, R 5 -CH=CH-X P (and) X P is a phosphate group (e.g., a protected phosphate group); However, R 22 and R 23 One of them is the subsequent binding to the nucleotide-nucleotide bond to the nucleoside, R 22 and R 23 Only one of these is subsequent binding to the nucleotide-nucleotide bond of the nucleoside. Oligonucleotides containing at least one nucleoside

[0323] Embodiment 2:X S An oligonucleotide of Embodiment 1, wherein is O or CH2.

[0324] Embodiment 3:X S An oligonucleotide of any of Embodiments 1 to 2, wherein is O.

[0325] Embodiment 4:R 23 An oligonucleotide of any of Embodiments 1 to 3, wherein the subsequent binding is to the internucleotide bond to the nucleoside.

[0326] Embodiment 5: R 22 C is substituted with hydroxyl, protected hydroxyl, halogen, or optionally. 1-30Alkoxy (e.g., methoxy, 2-methoxyethoxy), alkoxyalkyl (e.g., 2-methoxyethyl), hydrogen, optionally substituted C 1-30 Alkyl, optionally substituted C 2-30 Alkenyl, optionally substituted with C 2-30 Alkynyl, alkoxyalkylamine, alkoxyoxycarboxylate, amino, alkylamino, dialkylamino, 5-8 membered heterocyclyl, -OC 4-30 Alkyl-ON(CH2R 8 )(CH2R 9 ), or -OC 4-30 Alkyl-ON(CH2R 8 )(CH2R 9 ) and; or R 24 and R 25 These become one 4'-C(R 10 R 11 ) v -Y-2' or 4'-YC(R 10 R 11 ) v The oligonucleotide of embodiment 4, which is -2'.

[0327] Embodiment 6:R 22 C is substituted with hydroxyl, protected hydroxyl, halogen, or optionally. 1-30 The elements are alkoxy (e.g., methoxy, 2-methoxyethoxy), alkoxyalkyl (e.g., 2-methoxyethyl), hydrogen, amino, alkylamino, or dialkylamino; or R 22 and R 24 These become one 4'-C(R 10 R 11 ) v -Y-2' or 4'-YC(R 10 R 11 ) v An oligonucleotide of any of embodiments 4 to 5, which is -2'.

[0328] Embodiment 7:R 22 is hydrogen, hydroxyl, protected hydroxyl, fluoro, methoxy, ethoxy, or 2-methoxyethoxy; or R22 and R 24 These become one 4'-C(R 10 R 11 ) v -Y-2' or 4'-YC(R 10 R 11 ) v An oligonucleotide of any of embodiments 4 to 6, which is -2'.

[0329] Embodiment 8:R 22 An oligonucleotide of any of Embodiments 4 to 7, wherein is hydrogen, hydroxyl, protected hydroxyl, fluoro, or methoxy.

[0330] Embodiment 9:R 22 and R 24 These become one 4'-C(R 10 R 11 ) v -Y-2' or 4'-YC(R 10 R 11 ) v An oligonucleotide of any of embodiments 4 to 7, which is -2'.

[0331] Embodiment 10:R 2 and R 4 These become one 4'-C(R 10 R 11 ) v The oligonucleotide of Embodiment 9 is -Y-2', where v is 1 or 2.

[0332] Embodiment 11:R 10 and R 11 Oligonucleotides of Embodiment 9 or 10, wherein one of the atoms is H and the other is independently H or a C1-C6 alkyl group which is optionally substituted.

[0333] Embodiment 12:R 22 and R 24 The oligonucleotide of embodiment 11, in which the two components combine to form 4'-CH2-O-2'.

[0334] Embodiment 13:R 24An oligonucleotide of any of embodiments 4 to 8, wherein the element is H.

[0335] Embodiment 14:R 22 An oligonucleotide of any of Embodiments 1 to 3, wherein the subsequent binding is to the internucleotide bond to the nucleoside.

[0336] Embodiment 15:R 23 C is substituted with hydroxyl, protected hydroxyl, halogen, or optionally. 1-30 Alkoxy (e.g., methoxy, 2-methoxyethoxy), alkoxyalkyl (e.g., 2-methoxyethyl), hydrogen, optionally substituted C 1-30 Alkyl, optionally substituted C 2-30 Alkenyl, optionally substituted with C 2-30 Alkynyl, alkoxyalkylamine, alkoxyoxycarboxylate, amino, alkylamino, dialkylamino, 5-8 membered heterocyclyl, -OC 4-30 Alkyl-ON(CH2R 8 )(CH2R 9 ), or -OC 4-30 Alkyl-ON(CH2R 8 )(CH2R 9 ) an oligonucleotide of embodiment 14.

[0337] Embodiment 16:R 23 C is substituted with hydroxyl, protected hydroxyl, halogen, or optionally. 1-30 An oligonucleotide of any of Embodiments 14-15, which is an alkoxy (e.g., methoxy, 2-methoxyethoxy), an alkoxyalkyl (e.g., 2-methoxyethyl), hydrogen, amino, alkylamino, or dialkylamino.

[0338] Embodiment 17:R 23 is hydrogen, hydroxyl, protected hydroxyl, fluoro, methoxy, ethoxy, or 2-methoxyethoxy; or R 2 and R 4 These become one 4'-C(R 10 R11 ) v -Y-2' or 4'-YC(R 10 R 11 ) v An oligonucleotide of any of embodiments 14 to 16, which is -2'.

[0339] Embodiment 18:R 23 An oligonucleotide of any of embodiments 14 to 17, wherein is hydrogen, hydroxyl, protected hydroxyl, fluoro, or methoxy.

[0340] Embodiment 19:R 24 An oligonucleotide of any of embodiments 14 to 18, wherein the base molecule is H.

[0341] Embodiment 20: The nucleotide of formula (A) is of formula (A-VP) or (A-VP'): [ka] An oligonucleotide of any of embodiments 1 to 18.

[0342] Embodiment 21:R 23 The oligonucleotide of Embodiment 20 is a subsequent binding to the internucleotide bond of the nucleotide.

[0343] Embodiment 22:R 22 Oligonucleotides of Embodiment 21, wherein is hydrogen, hydroxyl, protected hydroxyl, fluoro, or methoxy.

[0344] Embodiment 23:X P One of them is -P(O)(OR V )2, where each R V An oligonucleotide of any of embodiments 20 to 22, wherein is independently H or an oxygen protecting group.

[0345] Embodiment 24: Each R V An oligonucleotide of embodiment 23, wherein H is independently present.

[0346] Embodiment 25:X P -P(O)(OR V )2, R 23 This is the subsequent binding to the internucleotide bond of the nucleotide, R 22 R is hydrogen, hydroxyl, protected hydroxyl, fluoro, or methoxy, where each R V Oligonucleotides of Embodiment 20, wherein the group is independently H or an oxygen protecting group.

[0347] Embodiment 26: Each R V An oligonucleotide of embodiment 25, wherein H is independently present.

[0348] Embodiment 27:R 22 The oligonucleotide of Embodiment 20 is a subsequent binding to the internucleotide bond of the nucleotide.

[0349] Embodiment 28:R 23 Oligonucleotides of Embodiment 27, wherein is hydrogen, hydroxyl, protected hydroxyl, fluoro, or methoxy.

[0350] Embodiment 29:X P -P(O)(OR V )2, where each R V Oligonucleotides of embodiment 27 or 28, wherein the group is independently H or an oxygen protecting group.

[0351] Embodiment 30: Each R V An oligonucleotide of embodiment 29, wherein H is independently present.

[0352] Embodiment 31:X P -P(O)(OR V )2, R 22 This is the subsequent binding to the internucleotide bond of the nucleotide, R 23 R is hydrogen, hydroxyl, protected hydroxyl, fluoro, or methoxy, where each R V Oligonucleotides of Embodiment 20, wherein the group is independently H or an oxygen protecting group.

[0353] Embodiment 32: Each R V An oligonucleotide of embodiment 31 in which is independently H.

[0354] Embodiment 33: An oligonucleotide of any of Embodiments 1 to 32, wherein the oligonucleotide contains 3 to 50 nucleotides.

[0355] Embodiment 34: An oligonucleotide of any of Embodiments 1 to 33, wherein the oligonucleotide comprises at least one ribonucleotide.

[0356] Embodiment 35: An oligonucleotide of any of Embodiments 1 to 34, wherein the oligonucleotide comprises at least one 2'-deoxyribonucleotide.

[0357] Embodiment 36: An oligonucleotide of any of Embodiments 1 to 35, wherein the oligonucleotide comprises, in addition to the nucleoside of formula (A), at least one nucleoside having a modified or non-native nucleic acid base.

[0358] Embodiment 37: An oligonucleotide of any of Embodiments 1 to 36, wherein the oligonucleotide comprises a nucleoside of formula (A) in addition to at least one nucleoside having a modified ribose sugar.

[0359] Embodiment 38: An oligonucleotide of any of Embodiments 1 to 37, wherein the oligonucleotide comprises, in addition to the nucleoside of formula (A), at least one nucleoside having a group other than H or OH at the 2' position of the ribose sugar.

[0360] Embodiment 39: An oligonucleotide of any of Embodiments 1 to 38, wherein the oligonucleotide comprises a nucleoside of formula (A) in addition to at least one nucleoside having a 2'-F ribose.

[0361] Embodiment 40: An oligonucleotide of any of Embodiments 1 to 39, wherein the oligonucleotide comprises a nucleoside of formula (A) in addition to at least one nucleoside having a 2'-OMe ribose.

[0362] Embodiment 41: An oligonucleotide of any of Embodiments 1 to 40, wherein the oligonucleotide comprises at least one nucleoside containing a portion other than the ribose sugar, in addition to the nucleoside of formula (A).

[0363] Embodiment 42: An oligonucleotide of any of Embodiments 1 to 41, wherein the oligonucleotide comprises at least one modified nucleotide interbonding.

[0364] Embodiment 43: Any oligonucleotide of Embodiments 1 to 42, wherein the subsequent internucleotide bond to the nucleoside is a modified internucleotide bond.

[0365] Embodiment 44: The oligonucleotide of Embodiment 43, wherein the bond between modified nucleotides is a phosphorothioate bond.

[0366] Embodiment 45: An oligonucleotide of any of Embodiments 1 to 44, wherein the oligonucleotide is bound to a solid support.

[0367] Embodiment 46: An oligonucleotide of any of Embodiments 1 to 45, wherein the oligonucleotide comprises at least one ligand.

[0368] Embodiment 47: An oligonucleotide of any of Embodiments 1 to 46, wherein the oligonucleotide comprises at least one hydroxyl, phosphate, or amino protecting group.

[0369] Embodiment 48: A double-stranded nucleic acid comprising a first oligonucleotide chain and a second oligonucleotide chain substantially complementary to the first chain, wherein the first or second chain is an oligonucleotide of any of Embodiments 1 to 47.

[0370] Embodiment 49: A double-stranded nucleic acid of Embodiment 48, wherein one of the first and second strands is an oligonucleotide from any of Embodiments 1 to 47.

[0371] Embodiment 50: A double-stranded nucleic acid of Embodiment 48 or 50, wherein the first and second strands are independently 15 to 25 nucleotides long.

[0372] Embodiment 51: A double-stranded nucleic acid of any of Embodiments 48-50, which can induce RNA interference.

[0373] Embodiment 52: The double-stranded nucleic acid of Embodiment 51, wherein the double-stranded nucleic acid comprises an antisense strand and a sense strand, wherein the antisense strand is an oligonucleotide of any of Embodiments 1 to 47.

[0374] Embodiment 53: Any double-stranded nucleic acid of Embodiments 48-52, wherein one or both strands have 1-5 nucleotide overhangs at their respective 5' or 3' ends.

[0375] Embodiment 54: A double-stranded nucleic acid of any of Embodiments 48-53, wherein the fruit of one strand has a 2-nucleotide overhang at the 5' or 3' end.

[0376] Embodiment 55: A double-stranded nucleic acid of any of Embodiments 48-54, wherein the fruit of one strand has a 2-nucleotide overhang at its 3' end.

[0377] Embodiment 56: A pharmaceutical composition comprising any oligonucleotide from Embodiments 1 to 47 or any dsRNA molecule from Embodiments 48 to 55, either alone or in combination with a pharmaceutically acceptable carrier or additive.

[0378] Embodiment 57: A gene silencing kit comprising any oligonucleotide from Embodiments 1 to 47 or any dsRNA molecule from Embodiments 48 to 55.

[0379] Embodiment 58: A method for silencing a target gene in cells, comprising the step of introducing into cells (i) any double-stranded RNA of Embodiments 48-55 (where the antisense strand contains a nucleotide sequence substantially complementary to the target gene); or (ii) any oligonucleotide of Embodiments 1-47 (where the oligonucleotide contains a nucleotide sequence substantially complementary to the target gene).

[0380] Embodiment 59: A method for reducing the expression of a target gene in a subject, comprising administering to the subject (i) any double-stranded RNA of Embodiments 48-55 (wherein the antisense strand comprises a nucleotide sequence substantially complementary to the target gene); or (ii) any oligonucleotide of Embodiments 1-47 (wherein the oligonucleotide comprises a nucleotide sequence substantially complementary to the target gene).

[0381] Embodiment 60: The method of Embodiment 59, wherein the administration is subcutaneous or intravenous.

[0382] Embodiment 61: Formula (B): [ka] (Formula B) [During the ceremony: B is a nucleic acid base that is modified as desired; X S is O, CH2, S, or NH; R 2 C is substituted with hydroxyl, protected hydroxyl, halogen, or optionally. 1-30 Alkoxy (e.g., methoxy, 2-methoxyethoxy), alkoxyalkyl (e.g., 2-methoxyethyl), hydrogen, optionally substituted C 1-30 Alkyl, optionally substituted C 2-30 Alkenyl, optionally substituted with C 2-30 Alkynyl, alkoxyalkylamine, alkoxyoxycarboxylate, amino, alkylamino, dialkylamino, 5-8 membered heterocyclyl, -OC4-30 Alkyl-ON(CH2R 8 )(CH2R 9 ), -OC 4-30 Alkyl-ON(CH2R 8 )(CH2R 9 ), a phosphate group, a reactive phosphorus group, a ligand, or a linker covalently bonded to one or more ligands; R 3 C is a reactive phosphorus group, hydroxyl, protected hydroxyl, halogen, or optionally substituted C 2-30 Alkinyl, optionally substituted with C 1-30 Alkoxy (e.g., methoxy, 2-methoxyethoxy), alkoxyalkyl (e.g., 2-methoxyethyl), hydrogen, optionally substituted C 1-30 Alkyl, optionally substituted C 2-30 Alkenyl, alkoxyalkylamine, alkoxyoxycarboxylate, amino, alkylamino, dialkylamino, 5-8 membered heterocyclyl, -OC 4-30 Alkyl-ON(CH2R 8 )(CH2R 9 ), -OC 4-30 Alkyl-ON(CH2R 8 )(CH2R 9 ), a phosphate group, a ligand, or a linker covalently bonded to one or more ligands; R 4 C is substituted with hydrogen, if desired. 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted with C 2-6 Alkinyl, or C as desired. 1-6 It is an alkoxy; or R 4 and R 2 They become one 4'-C(R 10 R 11 ) v -Y-2' or 4'-YC(R 10 R 11 ) v -2' is; Y is -O-, -CH2-, -CH(Me)-, -C(CH3)2-, -S-, -N(R 12 )-, -C(O)-, -C(S)-, -S(O)-, -S(O)2-, -OC(O)-, -C(O)O-, -N(R 12 )C(O)-, or -C(O)N(R 12 )-and; R 10 and R 11 These are independently H, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, or optionally substituted C2-C6 alkynyl; R 12 C is substituted with hydrogen, if desired. 1-30 Alkyl, optionally substituted C1-C 30 Alkoxy, C 1-4 Haloalkyl, optionally substituted C 2-4 Alkenyl, optionally substituted with C 2-4 Alkinyl, optionally substituted with C 1-30 It is an alkyl-CO2H or nitrogen-protecting group; v is 1, 2, or 3; and R 5 ha-L 5 -CH=CH-X P And, Here, L 5 is a combination or C 1-30 It is an alkylene (for example, L 5 It is a bond, that is, R 5 -CH=CH-X P (and) X P is a phosphate group (for example, a protected phosphate group), However, R 2 and R 3 When one of them is a reactive phosphorus group, R 2 and R 3 Only one of them is a reactive phosphorus group. A compound of [this].

[0383] Embodiment 62:X SA compound of embodiment 61, wherein is O or CH2.

[0384] Embodiment 63:X S A compound of any of embodiments 61 to 62, wherein is O.

[0385] Embodiment 64:R 3 A compound of any of embodiments 61 to 63, wherein is a reactive phosphorus group, a hydroxyl group, or a protected hydroxyl group.

[0386] Embodiment 65:R 3 The compound of embodiment 64, wherein is a reactive phosphorus group.

[0387] Embodiment 66:R 2 C is substituted with hydroxyl, protected hydroxyl, halogen, or optionally. 1-30 Alkoxy (e.g., methoxy, 2-methoxyethoxy), alkoxyalkyl (e.g., 2-methoxyethyl), hydrogen, optionally substituted C 1-30 Alkyl, optionally substituted C 2-30 Alkenyl, optionally substituted with C 2-30 Alkynyl, alkoxyalkylamine, alkoxyoxycarboxylate, amino, alkylamino, dialkylamino, 5-8 membered heterocyclyl, -OC 4-30 Alkyl-ON(CH2R 8 )(CH2R 9 ), or -OC 4-30 Alkyl-ON(CH2R 8 )(CH2R 9 ) and; or R 2 and R 4 These become one 4'-C(R 10 R 11 ) v -Y-2' or 4'-YC(R 10 R 11 ) v A compound of any of the embodiments 64 to 65, which is -2'.

[0388] Embodiment 67:R 2C is substituted with hydroxyl, protected hydroxyl, halogen, or optionally. 1-30 The elements are alkoxy (e.g., methoxy, 2-methoxyethoxy), alkoxyalkyl (e.g., 2-methoxyethyl), hydrogen, amino, alkylamino, or dialkylamino; or R 2 and R 4 These become one 4'-C(R 10 R 11 ) v -Y-2' or 4'-YC(R 10 R 11 ) v A compound of any of embodiments 64 to 66, which is -2'.

[0389] Embodiment 68:R 2 is hydrogen, hydroxyl, protected hydroxyl, fluoro, methoxy, ethoxy, or 2-methoxyethoxy; or R 2 and R 4 These become one 4'-C(R 10 R 11 ) v -Y-2' or 4'-YC(R 10 R 11 ) v A compound of any of embodiments 64 to 67, which is -2'.

[0390] Embodiment 69:R 2 A compound of any of embodiments 64 to 68, wherein is hydrogen, hydroxyl, protected hydroxyl, fluoro, or methoxy.

[0391] Embodiment 70:R 2 and R 4 These become one 4'-C(R 10 R 11 ) v -Y-2' or 4'-YC(R 10 R 11 ) v A compound of any of embodiments 64 to 69, which is -2'.

[0392] Embodiment 71:R 2 and R 4 These become one 4'-C(R10 R 11 ) v The compound of embodiment 70, which is -Y-2', where v is 1 or 2.

[0393] Embodiment 72:R 10 and R 11 A compound of Embodiment 70 or 71, wherein one of the atoms is H and the other is independently H or optionally substituted C1-C6 alkyl.

[0394] Embodiment 73:R 2 and R 4 The compound of embodiment 72, in which the two components combine to form 4'-CH2-O-2'.

[0395] Embodiment 74:R 4 A compound of any of embodiments 64 to 69, wherein is H.

[0396] Embodiment 75:R 2 A compound of any of embodiments 61 to 63, wherein is a reactive phosphorus group, a hydroxyl group, or a protected hydroxyl group.

[0397] Embodiment 76:R 2 The compound of embodiment 75, wherein is a reactive phosphorus group.

[0398] Embodiment 77:R 3 C is substituted with hydroxyl, protected hydroxyl, halogen, or optionally. 1-30 Alkoxy (e.g., methoxy, 2-methoxyethoxy), alkoxyalkyl (e.g., 2-methoxyethyl), hydrogen, optionally substituted C 1-30 Alkyl, optionally substituted C 2-30 Alkenyl, optionally substituted with C 2-30 Alkynyl, alkoxyalkylamine, alkoxyoxycarboxylate, amino, alkylamino, dialkylamino, 5-8 membered heterocyclyl, -OC 4-30 Alkyl-ON(CH2R 8 )(CH2R 9 ), or -OC 4-30Alkyl-ON(CH2R 8 )(CH2R 9 A compound of any of the embodiments 75 to 76.

[0399] Embodiment 78:R 3 C is substituted with hydroxyl, protected hydroxyl, halogen, or optionally. 1-30 A compound of any of embodiments 75 to 77, which is an alkoxy (e.g., methoxy, 2-methoxyethoxy), an alkoxyalkyl (e.g., 2-methoxyethyl), hydrogen, amino, alkylamino, or dialkylamino.

[0400] Embodiment 79:R 3 is hydrogen, hydroxyl, protected hydroxyl, fluoro, methoxy, ethoxy, or 2-methoxyethoxy; or R 2 and R 4 These become one 4'-C(R 10 R 11 ) v -Y-2' or 4'-YC(R 10 R 11 ) v A compound of any of embodiments 75 to 78, which is -2'.

[0401] Embodiment 80:R 3 A compound of any of embodiments 75 to 79, wherein is hydrogen, hydroxyl, protected hydroxyl, fluoro, or methoxy.

[0402] Embodiment 81:R 4 A compound of any of embodiments 75 to 80, wherein is H.

[0403] Embodiment 82: The compound is of formula B-VP) or (B-VP'): [ka] The compound of embodiment 61.

[0404] Embodiment 83:R 3The compound of Embodiment 82, wherein is a reactive phosphorus group (e.g., a phosphoramidite, e.g., 3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite, 3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite, or 3'-[(β-thiobenzoylethyl)-(1-pyrrolidinyl)]-thiophosphoramidite).

[0405] Embodiment 84:R 2 The compound of Embodiment 83, wherein is hydrogen, hydroxyl, protected hydroxyl, fluoro, or methoxy.

[0406] Embodiment 85:X P One of them is -P(O)(OR V )2, where each R V A compound of any of embodiments 82 to 84, wherein is independently H or an oxygen protecting group.

[0407] Embodiment 86: Each R V The compound of embodiment 85, wherein is independently H.

[0408] Embodiment 87:X P -P(O)(OR V )2; R 3 a is a reactive phosphorus group (e.g., phosphoramidite, e.g., 3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite, 3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite, or 3'-[(β-thiobenzoylethyl)-(1-pyrrolidinyl)]-thiophosphoramidite); and R 2 The compound of Embodiment 82, wherein is hydrogen, hydroxyl, protected hydroxyl, fluoro, or methoxy.

[0409] Embodiment 88: Each R V The compound of embodiment 87, wherein is independently an oxygen protecting group.

[0410] Embodiment 89:R 2The compound of Embodiment 82, wherein is a reactive phosphorus group (e.g., a phosphoramidite, e.g., 3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite, 3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite, or 3'-[(β-thiobenzoylethyl)-(1-pyrrolidinyl)]-thiophosphoramidite).

[0411] Embodiment 90:R 3 The compound of Embodiment 89, wherein is hydrogen, hydroxyl, protected hydroxyl, fluoro, or methoxy.

[0412] Embodiment 91:X P -P(O)(OR V )2, where each R V Compounds of embodiment 89 or 90, wherein is independently H or an oxygen protecting group.

[0413] Embodiment 92: Each R V A compound of embodiment 91 in which is independently H.

[0414] Embodiment 93:X P -P(O)(OR V )2; R 2 is a reactive phosphorus group (e.g., phosphoramidite, e.g., 3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite, 3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite, or 3'-[(β-thiobenzoylethyl)-(1-pyrrolidinyl)]-thiophosphoramidite); and R 3 The compound of Embodiment 82, wherein is hydrogen, hydroxyl, protected hydroxyl, fluoro, or methoxy.

[0415] Embodiment 94: Each R V The compound of embodiment 93, wherein is independently an oxygen protecting group.

[0416] Embodiment 95: The compound [ka] The compound of Embodiment 1.

[0417] In a nucleotide of formula (A) or a compound of formula (B), R 5 -CH=CH-X P That is. R 5 -CH=CH-X P It should be noted that in this case, the double bond can be in a cis or trans configuration. Therefore, in some embodiment of any of the aspects described herein, R 5 -CH=CH-X P The double bond is in a cis configuration. In any other embodiment of any of the embodiments described herein, R 5 -CH=CH-X P The double bond is in a cis configuration.

[0418] Hydroxyl protecting group Various embodiments described herein include a hydroxyl protecting group. The hydroxyl protecting group is -R OP1 , -N(R OP2 )2, -C(=O)SR OP1 -C(=O)R OP1 , -CO2R OP1 -C(=O)N(R OP2 )2, -C(=NR OP2 )R OP1 -C(=NR OP2 )OR OP1 -C(=NR OP2 )N(R OP2 )2, -CH2OC(=O)R OP1 -S(=O)R OP1 , -SO2R OP1 , -Si(R OP1 )3, -P(R OP3 )2, -P(R OP3 ) + 3X - , -P(OR OP3 )2, -P(OR OP3 )3X - -P(=O)(R OP1 )2, -P(=O)(OR OP3 )2, and -P(=O)(N(ROP2 )2)2 including, but not limited to; here, each X - each R is a counterion; OP1 C is independent 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenil, C 2-10 Alkinyl, HeteroC 1-10 Alkyl, hetero C 2-10 Alkenyl, HeteroC 2-10 Alkinyl, C 3-10 Carbocyclyl, 3-14 member heterocyclyl, C 6-14 It is an aryl, or a 5- to 14-membered heteroaryl, or two R OP1 The groups unite to form a 3-14 member heteroreactive cyclic or 5-14 member heteroaryl ring; each R OP2 These are hydrogen, -OH, and -OR OP1 , -N(R OP3 )2, -CN, -C(=O)R OP1 -C(=O)N(R OP3 )2, -CO2R OP1 , -SO2R OP1 -C(=NR OP3 )OR OP1 -C(=NR OP3 )N(R OP3 )2, -SO2N(R OP3 )2, -SO2R OP3 , -SO2OR OP3 -SOR OP1 -C(=S)N(R OP3 )2, -C(=O)SR OP3 -C(=S)SR OP3 -P(=O)(R OP1 )2, -P(=O)(OR OP3 )2, -P(=O)(N(R OP3 )2)2, C 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenil, C 2-10 Alkinyl, HeteroC 1-10 Alkyl, hetero C 2-10 Alkenyl, HeteroC 2-10 Alkinyl, C 3-10Carbocyclyl, 3-14 member heterocyclyl, C 6-14 The aryl group is either a 5-14 member heteroaryl group or two R groups. OP2 The groups unite to form a 3-14 member heterocyclyl or 5-14 member heteroaryl ring; and each R OP3 Hydrogen and C are independent of each other. 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenil, C 2-10 Alkinyl, HeteroC 1-10 Alkyl, hetero C 2-10 Alkenyl, HeteroC 2-10 Alkinyl, C 3-10 Carbocyclyl, 3-14 member heterocyclyl, C 6-14 It is an aryl and a 5-14 member heteroaryl or two R OP3 The groups unite to form a 3-14 member heterocyclyl or 5-14 member heteroaryl ring; where R OP1 , R OP2 and R OP3 Each alkyl, alkenyl, alkynyl, carbocyryl, heterocyclyl, aralkyl, aryl, and heteroaryl are optionally OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, NH(C1-C4)alkyl, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8 )alkyl, O(C1-C8)alkyl (i.e., C1-C8 alkoxy), O(C1-C8)haloalkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) pIt may be substituted with one, two, three, four, or five substituents independently selected from -NH2 or CH2-aryl-alkoxy, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6.

[0419] Hydroxyl protecting groups are well known in this field and are incorporated herein by reference: Greene's Protecting Groups in Organic Synthesis, PGM Wuts, 5 th Includes details as described in Edition, John Wiley & Sons, 2014.

[0420] Exemplary hydroxyl protecting groups include methyl, t-butyloxycarbonyl (BOC or Boc), methoxymethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, pivaloyloxymethyl (POM), and acetyloxymethyl. Chil, (AM), 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-Gio Hoxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidine-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-Trichloroethyl, 2-Trimethylsilylethyl, 2-(phenylselenyl)ethyl, t-Butyl, Allyl, p-Chlorophenyl, p-Methoxyphenyl, 2,4-Dinitrophenyl, Benzyl (Bn), 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'-Dinitrobenzhydryl, 5-dibenzosberyl, 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(ruberinoyloxyphenyl)methyl, 4,4',4 "-Tris(benzoyloxyphenyl)methyl, 3-(imidazole-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-benzodisulfuran-2-yl, benzisothiazolyl, S-dioxide, trimethylsilyl (TMS), triethylsilyl (TES), triisopropyl Lucilyl (TIPS), Dimethylisopropylsilyl (IPDMS), Diethylisopropylsilyl (DEIPS), Dimethyltexylsilyl, t-Butyldimethylsilyl (TBDMS), t-Butyldiphenylsilyl (TBDPS), Tribenzylsilyl, Tri-p-Xylylsilyl, Triphenylsilyl, Diphenylmethylsilyl (DPMS), t-Butylmethoxyphenylsilyl (TBMPS), Formyl, Acetyl, Chloroacetyl, Dichloroacetyl, Trichloro Roacetyl, trifluoroacetyl, methoxyacetyl, triphenylmethoxyacetyl, phenoxyacetyl, p-chlorophenoxyacetyl, 3-phenylpropionyl, 4-oxopentanoyl(rebrinate), 4,4-(ethylenedithio)pentanoate(ruberinoyldithioacetal), adamantoate, crotonate, 4-methoxycrotonate, bezoate, p-phenylbezoate, 2,4,6-trimethylbezoate(mesitoate), alkyl methyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl 2,22-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 allyl 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-benzyl thiocarbonate, 4-ethoxy-1-naphthyl carbonate, methyl Dithiocarbonate, 2-iodobezoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)bezoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)bezoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinoate, (E)-2-methyl-2-butanoate, o-(methoxyacyl)bezoate, α-naphthoate, nitrate, alkyl This includes, but is not limited to, N,N,N',N'-tetramethylphosphodiamide, alkyl N-phenylcarbamate, borate, dimethylphosphinothionel, alkyl 2,4-dinitrophenyl sulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts).

[0421] In one embodiment, the hydroxyl protecting group is benzyl, benzoyl, 2,6-dichlorobenzyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, mesylate, tosylate, 4,4'-dimethoxytrityl (DMT), 9-phenylxanthin-9-yl (Pixyl), and 9-(p-methoxyphenyl)xanthin-9-yl (MOX). In one embodiment, the hydroxyl protecting group is selected from acetyl, benzyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, and dimethoxytrityl, where the more preferred hydroxyl protecting group is 4,4'-dimethoxytrityl. In one embodiment, the hydroxyl protecting group is pivaloyloxymethyl.

[0422] The terms "protected hydroxyl" and "protected hydroxyl" used here refer to formula -OR Pro It means the base of, and here, R Pro This is the oxygen protecting group as defined herein.

[0423] Amine protecting group Various embodiments described herein include amine protecting groups (also referred to here as amino protecting groups). Amine protecting groups are -OH, -OR NP1 , -N(R NP2 )2, -C(=O)R NP1 -C(=O)N(R NP2 )2, -CO2R NP1 , -SO2R NP1 -C(=NR NP2 )R NP1 -C(=NR NP2 )OR NP1 -C(=NR NP2 )N(R NP2 )2, -SO2N(R NP2 )2, -SO2R NP2 , -SO2OR NP2 -SOR NP1 -C(=S)N(R NP2 )2, -C(=O)SR NP2 -C(=S)SR NP2 , C 1-10 Alkyl (e.g., aralkyl, heteroaralkyl), C 2-10Alkenil, C 2-10 Alkinyl, C 3-10 Carbocyclyl, 3-14 member heterocyclyl, C 6-14 This includes, but is not limited to, aryl and 5-14 member heteroaryl groups, where each R NP1 C is independent 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenil, C 2-10 Alkinyl, HeteroC 1-10 Alkyl, hetero C 2-10 Alkenyl, HeteroC 2-10 Alkinyl, C 3-10 Carbocyclyl, 3-14 member heterocyclyl, C 6-14 It is an aryl, or a 5- to 14-membered heteroaryl, or two R NP1 The groups unite to form a 3-14 member heterocyclyl or 5-14 member heteroaryl ring; and each R NP2 Hydrogen and C are independent of each other. 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenil, C 2-10 Alkinyl, HeteroC 1-10 Alkyl, hetero C 2-10 Alkenyl, HeteroC 2-10 Alkinyl, C 3-10 Carbocyclyl, 3-14 member heterocyclyl, C 6-14 It is an aryl and a 5-14 member heteroaryl or two R SP3 The groups unite to form a 3-14 member heterocyclyl or 5-14 member heteroaryl ring, where R NP1 and R NP2Each alkyl, alkenyl, alkynyl, carbocyryl, heterocyclyl, aralkyl, aryl, and heteroaryl are optionally OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, NH(C1-C4)alkyl, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8 )alkyl, O(C1-C8)alkyl (i.e., C1-C8 alkoxy), O(C1-C8)haloalkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) p It may be substituted with one, two, three, four, or five substituents independently selected from -NH2 or CH2-aryl-alkoxy, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6.

[0424] Amine protecting groups are well known in this art and are incorporated herein by reference: Greene's Protecting Groups in Organic Synthesis, PGM Wuts, 5 th Includes details as described in Edition, John Wiley & Sons, 2014.

[0425] Exemplary amides (e.g., -C(=O)R) NP1)-based amine protecting groups include, but are not limited to, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolineamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivatives, benzamide, p-phenylbenzamide, o-nitophenylacetamide, o-nitrophenoxyacetamide, acetamide, (N'-dithiobenzyloxyacylamino)acetamide, 3-(p-hydroxylphenyl)propanamide, 3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide, 2-methyl-2-(o-phenylazofenoxy)propanamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine derivatives, o-nitrobenzamide, and o-(benzoyloxymethyl)benzamide.

[0426] Exemplary carbamates (e.g., -C(=O)OR) NP1The amine protecting groups based on ) include methyl carbamate, ethyl carbamate, 9-fluorenyl methyl carbamate (Fmoc), 9-(2-sulfo)fluorenyl methyl carbamate, 9-(2,7-dibromo)fluoroenyl methyl carbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxantyl)] methyl carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc), 2,2,2-trichloroethyl carbamate (Troc), and 2-trimethyl Silyl ethyl carbamate (Teoc), 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-butylphenyl)-1-methylethyl carbamate (t-Bu meoc), 2-(2'- and 4'-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamide)ethyl carbamate, t-butyl carbamate (BOC or 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-hydroxyl piperidinyl carbamate , alkyldithiocarbamate, benzylcarbamate (Cbz), p-methoxybenzylcarbamate (Moz), p-nitobenzylcarbamate, p-bromobenzylcarbamate, p-chlorobenzylcarbamate, 2,4-dichlorobenzylcarbamate, 4-methylsulfinylbenzylcarbamate (Msz), 9-anthrylmethylcarbamate, diphenylmethylcarbamate, 2-methylthioethylcarbamate, 2-methylsulfonylethylcarbamate, 2-(p-toluenesulfonyl)ethylcarbamate, [2-(1,3-Dithia(nyl))methylcarbamate (Dmoc), 4-methylthiophenylcarbamate (Mtpc), 2,4-dimethylthiophenylcarbamate (Bmpc), 2-phosphonioethylcarbamate (Peoc), 2-triphenylphosphonioisopropylcarbamate (Ppoc), 1,1-dimethyl-2-cyanoethylcarbamate, m-chloro-p-acyloxybenzylcarbamate, p-(dihydroxylboryl)benzylcarbamate, 5-benzoisoxazolylmethylcarbamate, 2-(trifluoromethyl)-6- Chromonyl methylcarbamate (Tcroc), m-nitrophenylcarbamate, 3,5-dimethoxybenzylcarbamate, o-nitrobenzylcarbamate, 3,4-dimethoxy-6-nitrobenzylcarbamate, phenyl(o-nitrophenyl)methylcarbamate, t-amylcarbamate, S-benzylthiocarbamate, p-cyanobenzylcarbamate, cyclobutylcarbamate, cyclohexylcarbamate, cyclopentylcarbamate, cyclopropylmethylcarbamate, p-decyloxybenzylcarbamate, 2 ,2-Dimethoxyacylvinylcarbamate, o-(N,N-dimethylcarboxamide)benzylcarbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamide)propylcarbamate, 1,1-dimethylpropynylcarbamate, di(2-pyridyl)methylcarbamate, 2-furanylmethylcarbamate, 2-iodoethylcarbamate, isobornylcarbamate, isobutylcarbamate, isonicotinylcarbamate, p-(p'-methoxyphenylazo)benzylcarbamate, 1-methylcyclobutylcarbamate, 1 -Methylcyclohexylcarbamate, 1-methyl-1-cyclopropylmethylcarbamate, 1-methyl-1-(3,5-dimethoxyphenyl)ethylcarbamate, 1-methyl-1-(p-phenylazophenyl)ethylcarbamate, 1-methyl-1-phenylethylcarbamate, 1-methyl-1-(4-pyridyl)ethylcarbamate, phenylcarbamate, p-(phenylazo)benzylcarbamate, 2,4,6-tri-t-butylphenylcarbamate, 4-(trimethylammonium)benzylcarbamate, and 2,4,This includes, but is not limited to, 6-trimethylbenzylcarbamate.

[0427] Exemplary sulfonamides (e.g., -S(=O)2R) NP1 Examples of amine protecting groups based on ) include 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), and 2,4,6-trimethylbenzenesulfonamide (Mt This includes, but is not limited to, s), 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.

[0428] Further exemplary amine protecting groups include phenothiazinyl-(10)-acyl derivatives, N'-p-toluenesulfonylaminoacyl derivatives, N'-phenylaminothioacyl derivatives, N-benzoylphenylalanyl derivatives, N-acetylmethionine derivatives, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, and N-1,1,4,4-tetramethyldisilyl Zacyclopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexane-2-one, 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexane-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-pyrroline-3-yl) Amines, quaternary ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosberylamine, 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-dimethylthio Methyleneamine, N-benzylideneamine, Np-methoxybenzylideneamine, 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-hydroxylphenyl)phenylmethyleneamine, N-cyclohexyllideneamine, N-(5,This includes, but is not limited to, 5-dimethyl-3-oxo-1-cyclohexenyl)amine, N-borane and N-diphenylboric acid derivatives, N-[phenyl(pentoNP1silcrhromium- or tungsten)acyl]amine, N-copper chelate, N-zinc chelate, N-nitroamine, 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, and 3-nitropyridinesulfenamide (Npys).

[0429] Thiol protecting group Various embodiments described herein include a thiol protecting group. The thiol protecting group is -R SP1 , -N(R SP2 )2, -C(=O)SR SP1 -C(=O)R SP1 , -CO2R SP1 -C(=O)N(R SP2 )2, -C(=NR SP2 )R SP1 -C(=NR SP2 )OR SP1 -C(=NR SP2 )N(R SP2 )2, -S(=O)R SP1 , -SO2R SP1 , -Si(R SP1 )3, -P(R SP3 )2, -P(R SP3 ) + 3X - , -P(OR SP3 )2, -P(OR SP3 ) + 3X - -P(=O)(R SP1 )2, -P(=O)(OR SP3 )2, and -P(=O)(N(RSP2 )2)2 includes, but is not limited to, X - each R is a counterion; SP1 C is independent 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenil, C 2-10 Alkinyl, HeteroC 1-10 Alkyl, hetero C 2-10 Alkenyl, HeteroC 2-10 Alkinyl, C 3-10 Carbocyclyl, 3-14 member heterocyclyl, C 6-14 It is an aryl, or a 5- to 14-membered heteroaryl, or two R SP1 The groups unite to form a 3-14 member heterocyclyl or 5-14 member heteroaryl ring; each R SP2 These are hydrogen, -OH, and -OR SP1 , -N(R SP3 )2, -CN, -C(=O)R SP1 -C(=O)N(R SP3 )2, -CO2R SP1 , -SO2R SP1 -C(=NR SP3 )OR SP1 -C(=NR SP3 )N(R SP3 )2, -SO2N(R SP3 )2, -SO2R SP3 , -SO2OR SP3 -SOR SP1 -C(=S)N(R SP3 )2, -C(=O)SR SP3 -C(=S)SR SP3 -P(=O)(R SP1 )2, -P(=O)(OR SP3 )2, -P(=O)(N(R SP3 )2)2, C 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenil, C 2-10 Alkinyl, HeteroC 1-10 Alkyl, hetero C 2-10 Alkenyl, HeteroC 2-10 Alkinyl, C 3-10 Carbocyclyl, 3-14 member heterocyclyl, C6-14 The aryl group is either a 5-14 member heteroaryl group or two R groups. SP2 The groups unite to form a 3-14 member heterocyclyl or 5-14 member heteroaryl ring; and each R SP3 Hydrogen and C are independent of each other. 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenil, C 2-10 Alkinyl, HeteroC 1-10 Alkyl, hetero C 2-10 Alkenyl, HeteroC 2-10 Alkinyl, C 3-10 Carbocyclyl, 3-14 member heterocyclyl, C 6-14 It is an aryl and a 5-14 member heteroaryl or two R SP3 The groups unite to form a 3-14 member heterocyclyl or 5-14 member heteroaryl ring; where R SP1 , R SP2 and R SP3 Each alkyl, alkenyl, alkynyl, carbocyryl, heterocyclyl, aralkyl, aryl, and heteroaryl are optionally OH, CN, SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, NH(C1-C4)alkyl, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8 )alkyl, O(C1-C8)alkyl (i.e., C1-C8 alkoxy), O(C1-C8)haloalkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)] m -(CH2) p -OH, CH2-[CH(OH)] m -(CH2) pIt may be substituted with one, two, three, four, or five substituents independently selected from -NH2 or CH2-aryl-alkoxy, where "m" and "p" are independently 1, 2, 3, 4, 5, or 6.

[0430] Sulfur protecting groups are well known in this field and are incorporated herein by reference: Greene's Protecting Groups in Organic Synthesis, PGM Wuts, 5 th Includes details as described in Edition, John Wiley & Sons, 2014.

[0431] definition For convenience, certain terms used in the specification, examples, and accompanying claims are set forth here. Unless otherwise specified or implied by the context, the following terms and phrases include the meanings provided below. Unless otherwise explicitly indicated or evident from the context, the following terms and phrases are not intended to exclude the meanings acquired by such terms or phrases in the art to which they belong. Definitions are provided to aid in the description of specific embodiments and are not intended to limit the claimed invention, as the scope of the invention is limited only by the claims. Furthermore, unless otherwise required by the context, singular terms include plural terms and plural terms include singular terms.

[0432] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which this invention pertains. Any known methods, devices, and materials may be used in carrying out or testing the present invention, but in this regard, methods, devices, and materials are described herein. Definitions of common terms in immunology and molecular biology are found in: The Merck Manual of Diagnosis and Therapy, 20th Edition, Merck Sharp & Dohme Corp., 2018 (ISBN 0911910190, 978-0911910421); Robert S. Porter et al. (eds.), The Encyclopedia of Molecular Cell Biology and Molecular Medicine, published by Blackwell Science Ltd., 1999-2012 (ISBN 978352760431); and Robert A. Meyers (ed.), Molecular Biology and Biotechnology: a Comprehensive Desk Reference, published by VCH Publishers, Inc., 1995 (ISBN 1-56081-569-8); Immunology by Werner Luttmann, published by Elsevier, 2006; Janeway's Immunobiology, Kenneth Murphy, Allan Mowat, Casey Weaver (eds.), WW Norton & Company, 2016 (ISBN 0815345054, 978-0815345053); Lewin's Genes XI, published by Jones & Bartlett Publishers, 2014 (ISBN-1449659055); Michael Richard Green and Joseph Sambrook, Molecular Cloning: A Laboratory Manual, 4th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA (2012) (ISBN 1936113414); Davis et al., Basic Methods in Molecular Biology, Elsevier Science Publishing, Inc., New York, USA (2012) (ISBN 044460149X); Laboratory Methods in Enzymology: DNA, Jon Lorsch (ed.) Elsevier, 2013 (ISBN 0124199542); Current Protocols in Molecular Biology (CPMB), Frederick M. Ausubel (ed.), John Wiley and Sons, 2014 (ISBN 047150338X, 9780471503385), Current Protocols in Protein Science (CPPS), John E. Coligan (ed.), John Wiley and Sons, Inc., 2005; and Current Protocols in This information can be found in Immunology (CPI) (John E. Coligan, ADA M Kruisbeek, David H Margulies, Ethan M Shevach, Warren Strobe, (eds.) John Wiley and Sons, Inc., 2003 (ISBN 0471142735, 9780471142737)), and all of this content is incorporated herein by reference.

[0433] Furthermore, unless otherwise specified, the implementation of the present invention will utilize conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry, and immunology, which are within the scope of the art of those skilled in the art. Such techniques are well described in literature such as “Molecular Cloning: A Laboratory Manual”, second edition (Sambrook et al., 1989); “Oligonucleotide Synthesis” (MJ Gait, ed., 1984); “Animal Cell Culture” (RI Freshney, ed., 1987); “Methods in Enzymology” (Academic Press, Inc.); “Current Protocols in Molecular Biology” (FM Ausubel et al., eds., 1987, and periodic updates); “PCR: The Polymerase Chain Reaction”, (Mullis et al., ed., 1994); “A Practical Guide to Molecular Cloning” (Perbal Bernard V., 1988); and “Phage Display: A Laboratory Manual” (Barbas et al., 2001).

[0434] When a range of values ​​is given, values ​​between the upper and lower limits of the range, and between each limit, and any other values ​​within the stated range, are included within the scope of the invention up to 1 / 10 of the lower limit unit, unless otherwise clearly indicated by the context. These smaller range conditions and lower limits are independently included in the smaller range and within the scope of the invention, and are subject to any particular exclusion from the stated range. When the stated range includes one or both limits, the range excluding either or both of these limits is also included in the invention.

[0435] Unless otherwise indicated except in the operational examples, all numbers representing amounts of components or reaction conditions used herein should be understood in all cases as being modified by the term “approximately.” When used in combination with a percentage, the term “approximately” means ±1%. In some embodiments of the various aspects described herein, when used in combination with a percentage, the term “approximately” means ±5%. The term “approximately” is used herein to provide literal support for the exact number that precedes it, as well as for numbers that the term is close to or approximates the preceding number. In determining whether a number is close to or approximates a particular number, any close or approximate number that is not listed may be a number that, in the context in which it is listed, is substantially equivalent to the particular number.

[0436] The terms "includes" or "contains" as used herein refer to compositions, methods, and their respective elements that are essential to the present invention, but also imply that there is still room for unspecified elements, whether essential or not.

[0437] The term "consists of" refers to the composition, method, and each of its elements described herein, excluding any elements not included in the description of the embodiment.

[0438] As used herein, the term "essentially consisting of" refers to elements necessary for a particular embodiment. This term allows for the presence of further elements that do not substantially affect the basic, novel, or functional features of that embodiment of the invention.

[0439] A singular expression includes plural subjects unless the context clearly indicates otherwise. Similarly, the term “or” includes “and” unless the context clearly indicates otherwise. It should be further noted that claims may be drafted with any optional elements excluded; that is, this statement is intended to act as an antecedent, using exclusionary terms such as “simply,” “only,” or “negative” limitations in relation to the description of the claimed elements.

[0440] The abbreviation "eg" originates from the Latin phrase "exempli gratia" and is used here to indicate an unrestricted example. Therefore, the abbreviation "eg" is synonymous with the term "for example."

[0441] The terms "siRNA" and "iRNA agent" used herein are interchangeable and refer to agents capable of intervening in the silencing of target RNA, such as mRNA, or transcripts of protein-coding genes. For convenience, such mRNA is also referred to here as the mRNA to be silenced. Such genes are also referred to as the target genes. Generally, the RNA to be silenced can be endogenous, exogenous, or pathogenic genes. Furthermore, RNA other than mRNA, such as tRNA and viral RNA, can also be targeted.

[0442] The term "RNAi intervention" used here refers to the ability to silence a target gene, such as mRNA, in a sequence-specific manner. While we do not wish to be constrained by theory, silencing is thought to utilize the RNAi mechanism or process and guide RNA, such as the antisense strand of dsRNA (where the antisense strand is 21-23 nucleotides long).

[0443] To the extent used herein and unless otherwise specified, the term “complementary,” when used in the description of a first nucleotide sequence relative to a second nucleotide sequence, means the ability of an oligonucleotide or polynucleotide containing a first nucleotide sequence to hybridize with an oligonucleotide or polynucleotide containing a second nucleotide sequence under certain conditions to form a double-stranded structure, as will be understood by those skilled in the art. Such conditions may be, for example, stringent conditions, where stringent conditions include: 400 mM NaCl, 40 mM PIPES, pH 6.4, 1 mM EDTA, 12–16 hours at 50°C or 70°C, followed by washing. Other conditions, such as physiologically relevant conditions that may be encountered in living organisms, may be applied. Those skilled in the art can determine the setting of the most appropriate conditions for testing the complementarity of the two sequences, according to the final application of the hybridized nucleotides.

[0444] As used herein with respect to a nucleotide sequence, “substantially complementary” means a nucleotide sequence having at least 80% identity between the substantially complementary nucleotide sequence and the exactly complementary sequence of the reference. For example, a nucleotide sequence having a percentage of identity of at least 85%, at least 90%, at least 93%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% (i.e., exactly complementary). Preferably, identity is assessed over a length of at least 15, for example, at least 16, at least 17, at least 18, at least 19, at least 20, or at least 21 nucleotides.

[0445] The terms "off-target" and "off-target effect" refer to any instance in which an effector molecule for a target causes an unintended effect through direct or indirect interaction with other target sequences, DNA sequences, cellular proteins, or other parts. For example, an "off-target effect" may occur when there is simultaneous degradation of another transcript due to partial homology or complementarity between the sense and / or antisense strands of the siRNA and the other transcript.

[0446] The terms “reduction,” “decrease,” “decrease,” or “inhibition” are all used here to mean a statistically significant reduction. In some embodiments, “decrease,” “decrease,” or “decrease” or “inhibition” typically means a reduction of at least 10% compared to a reference level (e.g., in the absence of a certain treatment or drug), and may include reductions of, for example, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or more. As used herein, “decrease” or “inhibition” does not mean complete inhibition or reduction compared to a reference level. “Complete inhibition” is 100% inhibition compared to a reference level. The reduction may preferably be to a level that is acceptable as being within the normal range for an individual without a disorder.

[0447] The terms “increase,” “boost,” “enhance,” or “activation” are all used here to mean an increase of a statistically significant amount. In one embodiment, the terms “increase,” “boost,” or “activation” could mean an increase of at least 10% compared to a reference level, e.g., at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90%, or up to 100% increase (including this), or any increase of 10–100% compared to a reference level, or at least about 2 times, or at least about 3 times, or at least about 4 times, or at least about 5 times, or at least about 10 times increase, or 2–10 times or more increase compared to a reference level. In the context of markers or symptoms, “increase” is a statistically significant increase of such a level.

[0448] In this context, the "end" of a chain refers to the first position from the nearest end. For example, the 5' end is the first position from the 5' end of the chain. Similarly, the 3' end is the first position from the 3' end of the chain.

[0449] In this context, the "terminal region" of a chain refers to positions 1 through 4, counting from the nearest end of the chain, for example, positions 1, 2, 3, and 4. For example, the 5' terminal region refers to positions 1 through 4, counting from the 5' end of the chain, for example, positions 1, 2, 3, and 4. Similarly, the 3' terminal region refers to positions 1 through 4, counting from the 3' end of the chain, for example, positions 1, 2, 3, and 4.

[0450] For example, the 5'-terminal region of the antisense chain is positions 1, 2, 3, and 4, counting from the 5' end of the antisense chain. Preferred 5'-terminal regions of the antisense chain are positions 1, 2, and 3, counting from the 5' end of the antisense chain. The 3'-terminal region of the antisense chain may be positions 1, 2, 3, and 4, counting from the 3' end of the chain. Preferred 3'-terminal regions of the antisense chain are positions 1, 2, and 3, counting from the 3' end of the antisense chain.

[0451] Similarly, the 5'-terminal region of the sense strand can be at positions 1, 2, 3, and 4, counting from the 5' end of the sense strand. Preferred 5'-terminal regions of the sense strand can be at positions 1, 2, and 3, counting from the 5' end of the sense strand. The 3'-terminal region of the sense strand can be at positions 1, 2, 3, and 4, counting from the 3' end of the strand. Preferred 3'-terminal regions of the sense strand can be at positions 1, 2, and 3, counting from the 3' end of the sense strand.

[0452] The "central region" of the chain used here refers to positions 5 through 17, counting from the 5' end of the chain, for example, positions 6 through 16, 6 through 15, 6 through 14, 6 through 13, 6 through 12, 7 through 15, 7 through 14, 7 through 13, 7 through 12, 8 through 16, 8 through 15, 8 through 14, 8 through 13, 8 through 12, 9 through 16, 9 through 15, 9 through 14, 9 through 13, 9 through 12, 10 through 16, 10 through 15, 10 through 14, 10 through 13, or 10 through 12. For example, the central region of the chain means positions 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 of the chain. The preferred central regions of the sense strand are positions 6, 7, 8, 9, 10, 11, 12, 13, and 14, counting from the 5' end of the sense strand. The more preferred central regions of the sense strand are positions 7, 8, 9, 10, 11, 12, and 13, counting from the 5' end of the sense strand. The preferred central regions of the antisense strand are positions 9, 10, 11, 12, 13, 14, 15, 16, and 17, counting from the 5' end of the antisense strand. The more preferred central regions of the antisense strand are positions 10, 11, 12, 13, 14, 15, and 16, counting from the 5' end of the antisense strand.

[0453] The term "in vitro" as used herein refers to events occurring in an artificial environment, such as a test tube or reaction vessel, or cell culture, rather than within a living organism (e.g., an animal or plant). The term "ex vivo" as used herein refers to cells that have been removed from a living organism and cultured outside of it (e.g., in a test tube). The term "in vivo" as used herein refers to events occurring within a living organism (e.g., an animal, plant, and / or microorganism).

[0454] As used herein, the terms “subject” or “patient” refer to any organism to which the compositions disclosed herein can be administered, for example, for experimental, diagnostic, and / or therapeutic purposes. Typical subjects include animals (e.g., mammals, e.g., mice, rats, rabbits, non-human primates, and humans) and / or plants. Typically, animals are vertebrates such as primates, rodents, domesticated or sporting animals. Primates include chimpanzees, crab-eating macaques, spider monkeys, and macaques, e.g., rhesus macaques. Rodents include mice, rats, woodchucks, ferrets, rabbits, and hamsters. Domesticated and sporting animals include cattle, horses, pigs, deer, bison, buffalo, felines, e.g., domestic cats, canines, e.g., dogs, foxes, wolves, birds, e.g., chickens, emus, ostriches, and fish, e.g., trout, catfish, and salmon. The patient or subject includes all of the above except for one or more subgroups or species, such as humans, primates, or rodents. In some embodiments of the aspects described herein, the subject is a mammal, such as a primate, such as a human. The terms “patient” and “subject” are used interchangeably here. The subject may be male or female.

[0455] Preferably, the subject is a mammal. Mammals may be, but are not limited to, humans, non-human primates, mice, rats, dogs, cats, horses, or cattle. Non-human mammals can be advantageously used as subjects representing animal models of human diseases and disorders. Furthermore, the compounds, compositions, and methods described herein may be used in livestock and / or pets.

[0456] The subjects may be those who have previously been diagnosed with or identified as having a condition requiring treatment. Alternatively, the subjects may also be those who have not been previously diagnosed. A “subject in need” for testing a particular condition may be a subject who has that condition, has been diagnosed with that condition, or is at risk of developing that condition.

[0457] In one embodiment, the subject is human. In another embodiment, the subject is an experimental animal or surrogate animal used as a disease model. This term does not imply a specific age or sex. Therefore, adult and neonatal subjects, as well as fetuses, whether male or female, are covered. Examples of subjects include humans, dogs, cats, cattle, goats, and mice. The term subject is intended to further include transgenic species. In one embodiment, the subject may be of European descent. In one embodiment, the subject may be of African American descent. In one embodiment, the subject may be of Asian descent.

[0458] In jurisdictions that prohibit patents for methods implemented in the human body, the meaning of “administering” a composition to a human subject is limited to a formulation of a regulated substance that the human subject self-administers by some technique (e.g., orally, by inhalation, topically, by injection, by insertion, etc.). The broadest reasonable interpretation is intended to conform to the laws or regulations defining patentable subject matter. In jurisdictions that do not prohibit patents for methods implemented in the human body, “administering” a composition includes methods of implementation in the human body and both the methods of such activity.

[0459] As used herein, "non-enteral administration" refers to administration via injection or intravenous infusion. Non-enteral administration includes, but is not limited to, subcutaneous, intravenous, or intramuscular administration.

[0460] The terms "subcutaneous administration" used here refer to administration directly beneath the skin. "Intravenous administration" refers to administration into a vein.

[0461] The term “dose” as used herein refers to a specific amount of a drug delivered in a single dose. In some embodiments, a dose may be administered in two or more pills, tablets, or injections. For example, in some embodiments, when subcutaneous administration is desired, the desired dose requires a volume that cannot be easily applied by a single injection. In such embodiments, the desired dose may be achieved using two or more injections. In some embodiments, a dose may be administered in two or more injections to minimize the individual’s injection site reaction.

[0462] The term "dosage unit" as used herein refers to the form in which the drug is provided. In one embodiment, the dosage unit is a vial containing a lyophilized antisense oligonucleotide. In another embodiment, the dosage unit is a vial containing a reconstituted antisense oligonucleotide.

[0463] The terms “treatment,” “to treat,” or “treatment of” (and their grammatical variations) mean that the severity of the condition in question is reduced, at least partially improved or stabilized, and / or that some degree of reduction, mitigation, reduction or stabilization of at least one clinical symptom is achieved, and / or the progression of the disease or disability is delayed.

[0464] The terms “prevention,” “prevent,” and “prevention” (and their grammatical variations) refer to the prevention and / or delay of the onset of disease, disability, and / or clinical symptoms in a subject and / or a reduction in the severity of the onset of disease, disability, and / or clinical symptoms compared to those occurring in the absence of the method of the present invention. Prevention may be complete, for example, the complete absence of disease, disability, and / or clinical symptoms. Prevention may also be the onset and / or reduction in the severity of the onset of disease, disability, and / or clinical symptoms in a subject compared to those occurring in the absence of the present invention.

[0465] The term "statistically significant" or "significant" refers to statistical significance, generally meaning a difference of two standard deviations (2SD) or more.

[0466] Glycolic acid nucleic acid (GNA) [ka] Here, B is a modified or unmodified nucleic acid base, and * is R, S, or racemic.

[0467] The term “acyclic nucleotide” refers to any nucleotide having an acyclic ribose sugar, for example, lacking any bonds between ribose carbons (e.g., C1'-C2', C2'-C3', C3'-C4', C4'-O4', or C1'-O4') and / or lacking at least one ribose carbon or oxygen (e.g., C1', C2', C3', C4', or O4') independently or in combination in the nucleotide. In one embodiment, an acyclic nucleotide is: [ka] Here, B is a modified or unmodified nucleic acid base, and R 1 and R 2 R3 is independently H, halogen, OR3, or alkyl; and R3 is H, alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, or sugar.

[0468] Unlocked nucleic acid (UNA) modifications include monomers having a bond in which the C1'-C4' bond (i.e., the covalent carbon-oxygen-carbon bond between the C1' and C4' carbons) has been removed. In other examples, the C2'-C3 bond (i.e., the covalent carbon-carbon bond between the C2 and C3 carbons) of the sugar is absent (see Mikhailov et al., Tetrahedron Letters, 26 (17): 2059 (1985); and Fluiter et al., Mol. Biosyst., 10: 1039 (2009), which are incorporated herein by reference as a whole). Acyclic nucleotides may be linked via 2'-5' or 3'-5' bonds. In one embodiment, the UNA structure is: [ka] The UNA modifier is a modified or unmodified nucleic acid base; R, R', R'', R''' and R'''' are independently H, OH, CH3, CH2CH3, O-alkyl, NH2, NHMe, or NMe2; and each * is independently R, S, or racemic. In one embodiment, the UNA modification is [ka] Here, B is a modified or unmodified nucleic acid base, and R is H, OH, or O-alkyl.

[0469] Modified Unlocked Nucleic Acid (mUNA) Modifications include the following: [ka] This includes, but is not limited to, the following, where a base is a modified or unmodified nucleic acid base, and * is R, S, or racemic.

[0470] In one embodiment, the UNA modification is selected from the group consisting of the following: [ka]

[0471] The term "debasic modification" refers to a nucleotide or its analogue that lacks a nucleic acid base. Some exemplary debasic modifications include: [ka] This includes, but is not limited to, R, where R is H, Me, Et or OMe; R' is H, Me, Et or OMe; R'' is H, Me, Et or OMe; and * is R, S or racemic.

[0472] In some cases, thermal destabilization modifications are: [ka] [In the formula, B is a modified or unmodified nucleic acid base, and * is R, S, or racemic.] It is selected from the group consisting of the following.

[0473] The 2'-5' RNA used here is [ka] Here, the base is either a modified or unmodified nucleic acid base.

[0474] The TNA used here is a nucleotide containing threose sugar instead of ribose sugar, where its 3' position is attached to the 3' position of its upstream nucleotide and its 2' position is attached to the 5' position of its downstream nucleotide. [ka] It has the following: Some exemplary TNA modifications are as follows: [ka] This includes, but is not limited to, the modified or unmodified nucleic acid bases.

[0475] The Hyp spacer modification used here is a hydroxyprolinol monomer, for example, [ka] This includes, where R is H or a modified or unmodified nucleic acid base.

[0476] Further exemplary sugar modifications include: [ka] This includes, but is not limited to, the following, where B is a modified or unmodified nucleic acid base and R is H or a C1-C6 alkyl (e.g., methyl or ethyl).

[0477] For example, nucleotides in which the WCH bond with the complementary base of the opposing chain is impaired include, but are not limited to, nucleotides containing nucleic acid bases independently selected from the following: [ka]

[0478] Exemplary non-canonical bases that have impaired or completely absent ability to form hydrogen bonds with a base in the opposing chain include, but are not limited to, inosine, nebularin, 2-aminopurine, 2,4-difluorotoluene, 5-nitroindole, 3-nitropyrrole, 4-fluoro-6-methylbenimidazole, and 4-methylbenzimidazole.

[0479] Exemplary α-nucleotides are, [ka] This includes, but is not limited to, B being a modified or unmodified nucleic acid base, and R being H, OH, OCH3, F, NH2, NHMe, NMe2, or O-alkyl.

[0480] Exemplary phosphate modifications known to reduce the thermal stability of dsNA double chains compared to natural phosphodiester bonds are as follows: [ka] This includes, but is not limited to, the alkyl group of the R group, which can be C1-C6 alkyl. Specific alkyl groups of the R group include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, pentyl, and hexyl.

[0481] The term "cross-linked nucleic acid" used herein refers to a nucleotide containing a five-membered or six-membered cross-linked structure with a fixed 3'-terminal confiscation, also known as a north confiscation. The cross-linked structure bonds the 2'-carbon (e.g., 2'-oxygen) of a ribose sugar to the 4'-carbon of a ribose sugar. Various different cross-linked structures are possible, including those containing carbon, oxygen, nitrogen, and hydrogen atoms.

[0482] In one embodiment, BNA is a locked nucleic acid (LNA). The term "locked nucleic acid" (LNA) as used here generally refers to a class of BNA in which the ribose ring is "locked" by a methylene bridge that connects the 2' oxygen of the ribose sugar to the 4' carbon of the ribose sugar.

[0483] As used herein, the term "aliphatic" refers to saturated or unsaturated and linear, branched, and / or cyclic hydrocarbons having a specified number of carbon atoms. Examples include alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, cycloalkylalkenyl, and cycloalkylalkynyl hydrocarbons having a specified number of carbon atoms.

[0484] The term "alkyl" as used herein refers to an aliphatic hydrocarbon group that may be linear or branched, having 1 to about 60 carbon atoms in the chain, preferably having about 6 to about 50 carbon atoms in the chain. "Lower alkyl" refers to an alkyl group having 1 to about 8 carbon atoms. "Higher alkyl" refers to an alkyl group having about 10 to about 20 carbon atoms. The alkyl group may be substituted with one or more alkyl substituents, which may be the same or different, and where "alkyl substituent" includes halo, amino, aryl, hydroxyl, alkoxy, aryloxy, alkyloxy, alkylthio, arylthio, aralkyloxy, aralkylthio, carboxy, alkoxycarbonyl, oxo, and cycloalkyl groups. "Branched" refers to an alkyl group in which a lower alkyl group, such as methyl, ethyl, or propyl, is bonded to a linear alkyl chain. Exemplary alkyl groups include methyl, ethyl, propyl, i-propyl, n-butyl, t-butyl, n-pentyl, hexyl, heptyl, octyl, decyl, dodecyl, tridecyl, tetradecyl, pentadecyl, and hexadecyl. Useful alkyl groups include branched or linear alkyl groups of 6 to 50 carbon atoms, as well as lower alkyl groups of 1 to about 4 carbon atoms and higher alkyl groups of about 12 to about 16 carbon atoms.

[0485] A "heteroalkyl" group is one in which a heteroatom, bonded to an appropriate number of hydrogen atoms, replaces one of the carbon atoms of an alkyl group (e.g., from a CH2 group to an NH group or an O group). The term "heteroalkyl" includes optionally substituted alkyl, alkenyl, and alkynyl groups having one or more skeletal chain atoms selected from atoms other than carbon, e.g., oxygen, nitrogen, sulfur, phosphorus, silicon, or combinations thereof. In some embodiments, the heteroatom is located at any internal position of the heteroalkyl group. Examples include, but are not limited to, -CH2-O-CH3, -CH2-CH2-O-CH3, -CH2-NH-CH3, -CH2-CH2-NH-CH3, -CH2-N(CH3)-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2,-S(O)-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=N-OCH3, and -CH=CH-N(CH3)-CH3. In some embodiments, up to two heteroatoms are consecutive, for example, -CH2-NH-OCH3 and -CH2-O-Si(CH3)3.

[0486] The term "alkenyl" as used herein refers to an alkyl group containing at least one carbon-carbon double bond. The alkenyl group may optionally be substituted with one or more "alkyl substituents." Exemplary alkenyl groups include vinyl, allyl, n-pentenyl, decenyl, dodecenyl, tetradecadienyl, heptadec-8-en-1-yl, and heptadec-8,11-dien-1-yl.

[0487] The term "alkynyl" as used herein refers to an alkyl group containing a carbon-carbon triple bond. The alkynyl group may optionally be substituted with one or more alkyl substituents. Exemplary alkynyl groups include ethynyl, propargyl, n-pentynyl, desinyl, and dodecynyl. Useful alkynyl groups include lower alkynyl groups.

[0488] The term "cycloalkyl" as used herein refers to a non-aromatic monocyclic or polycyclic ring system with approximately 3 to 12 carbon atoms. Cycloalkyl groups may optionally be partially unsaturated. Cycloalkyl groups may optionally be substituted with aryl substituents, oxo, and / or alkylenes. Representative monocyclic cycloalkyl rings include cyclopentyl, cyclohexyl, and cycloheptyl. Useful polycyclic cycloalkyl rings include adamantyl, octahydronaphthyl, decalin, camphor, camphan, and noadamantyl.

[0489] A "heterocyclyl" is a non-aromatic 3-8 member monocyclic, 8-12 member bicyclic, or 11-14 member tricyclic ring system having 1-3 heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic, where the heteroatoms are selected from O, N, or S (for example, if monocyclic, bicyclic, or tricyclic, each consisting of a carbon atom and 1-3, 1-6, or 1-9 N, O, or S heteroatoms, respectively). Cx heterocyclyls and Cx-Cy heterocyclyls are typically used, where X and Y represent the number of carbon atoms in the ring system. In some embodiments, one, two, or three hydrogen atoms in each ring may be substituted with substituents. Exemplary heterocyclyl groups include, but are not limited to, piperazinyl, pyrrolidinyl, dioxanyl, morpholinyl, tetrahydrofuranyl, piperidyl, 4-morpholyl, 4-piperazinyl, pyrrolidinyl, perhydropyrrolidinyl, 1,4-diazaperhydroepinyl, 1,3-dioxanyl, and 1,4-dioxanyl.

[0490] "Aryl" refers to an aromatic carbocyclic radical containing approximately 3 to 13 carbon atoms. The aryl group may be substituted with one or more identical or different aryl substituents, where "aryl substituent" includes alkyl, alkenyl, alkynyl, aryl, aralkyl, hydroxyl, alkoxy, aryloxy, aralkoxy, carboxy, aroyl, halo, nitro, trihalomethyl, cyano, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, acyloxy, acylamino, aroylamino, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, lylthio, alkylthio, alkylene, and -NRR', where R and R' are independently hydrogen, alkyl, aryl, and aralkyl, respectively. Exemplary aryl groups include substituted or unsubstituted phenyl and substituted or unsubstituted naphthyl.

[0491] A "heteroaryl" is an aromatic 3-8 member monocyclic, 8-12 member fused bicyclic, or 11-14 member fused tricyclic ring system having 1-3 heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic, and the heteroatoms are selected from O, N, or S (for example, if monocyclic, bicyclic, or tricyclic, each consisting of a carbon atom and 1-3, 1-6, or 1-9 heteroatoms of N, O, or S).

[0492] Exemplary aryls and heteroaryls include phenyl, pyridinyl, pyrimidinyl, furanyl, thienyl, imidazolyl, thiazolyl, pyrazolyl, pyridazinyl, pyrazinyl, triazinyl, tetrazolyl, indolyl, benzyl, naphthyl, anthracenyl, azurenyl, fluorenyl, indanyl, indenyl, naphthyl, tetrahydronaphthyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benz Soxazolyl, benzisothiazolyl, benzimidazolinil, carbazolyl, 4aH-carbazolyl, carborinil, chromanil, clomenil, cinnolinil, decahydroquinolinil, 2H,6H-1,5,2-dithiadinil, dihydrofloflo[2,3b]tetrahydrofuran, furanil, flazanil, imidazolidinil, imidazolinil, imidazolyl, 1H-indazolyl, indolinil, indolinil, indolininil, indolyl, 3H-indolyl, isatinoyl, isobenzofuranil, isochromanil, isoindazolyl, isoindolinil, Isoindolyl, Isoquinolinyl, Isothiazolyl, Isoxazolyl, Methylenedioxyphenyl, Morpholinyl, Naphthilidinyl, Octahydroisoquinolinyl, Oxadiazolyl, 1,2,3-Oxadiazolyl, 1,2,4-Oxadiazolyl, 1,2,5-Oxadiazolyl, 1,3,4-Oxadiazolyl, Oxazolidinyl, Oxazolyl, Oxoindolyl, Pyrimidinyl, Phenanthrolinyl, Phenanthrolinyl, Phenadinyl, Phenothiazinyl, Phenoxazinyl, Phenoxadinyl, Phthalazinyl, Piperadinyl , piperidonil, 4-piperidonil, piperonil, pteridinil, prinil, pyranil, pyrazinil, pyrazolidinil, pyrazolinil, pyrazolyl, pyridadinil, pyridoxazole, pyridoimidazole, pyridothiazole, pyridinil, pyridyl, pyrimidinil, pyrrolidinil, pyrrolinil, 2H-pyrrolyl, pyrrrolyl, quinazolinil, quinolinil, 4H-quinolidinil, quinoxalinil, quinuclidinil, tetrahydrofuranil, tetrahydroisoquinolinil, tetrahydroquinolinil, tetrazolyl, 6H-1,2,5-thiadiadinil, 1,2,This includes, but is not limited to, 3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, and xanthenyl. In some embodiments, one, two, three, or four hydrogen atoms in each ring may be substituted with substituents.

[0493] The terms "halogen" or "halo" used herein refer to atoms selected from fluorine, chlorine, bromine, and iodine. The terms "halogen radioactive isotope" or "halo isotope" refer to radioactive nuclides of atoms selected from fluorine, chlorine, bromine, and iodine.

[0494] As an isolated group or as part of a larger group, a "halogen-substituted moiety" or "halo-substituted moiety" means an aliphatic, alicyclic, or aromatic moiety, as defined herein, that is substituted with one or more "halo" atoms (as defined herein).

[0495] The term "haloalkyl" as used herein refers to alkyl and alkoxy structures having at least one substituent of fluorine, chlorine, bromine, or iodine, or a combination thereof. In embodiments in which the group contains more than one halogen, the halogens may be the same or different. The terms "fluoroalkyl" and "fluoroalkoxy" respectively include haloalkyl and haloalkoxy groups in which the halo is fluorine. Exemplary halo-substituted alkyls include haloalkyls, dihaloalkyls, trihaloalkyls, perhaloalkyls, etc. (For example, halo-substituted (C1-C3) alkyls include chloromethyl, dichloromethyl, difluoromethyl, trifluoromethyl (CF3), perfluoroethyl, 2,2,2-trifluoroethyl, 2,2,2-trifluoro-l,l-dichloroethyl, etc.).

[0496] The term "amino" as used herein means -NH2. The term "alkylamino" means a nitrogen moiety having one linear or branched unsaturated aliphatic, cyclyl, or heterocyclyl group bonded to nitrogen, e.g., -NH(alkyl). The term "dialkylamino" means a nitrogen moiety having two linear or branched unsaturated aliphatic, cyclyl, or heterocyclyl groups bonded to nitrogen, e.g., -N(alkyl)(alkyl). The term "alkylamino" includes "alkenylamino," "alkynylamino," "cyclylamino," and "heterocyclylamino." The term "arylamino" means a nitrogen moiety having at least one aryl group bonded to nitrogen, e.g., -NHaryl and -N(aryl)2. The term "heteroarylamino" means a nitrogen moiety having at least one heteroaryl group bonded to nitrogen, e.g., -NHheteroaryl and -N(heteroaryl)2. Optionally, two substituents may also form a ring with nitrogen. Unless otherwise specified, compounds containing an amino moiety listed herein include their protected derivatives. Suitable protecting groups for the amino moiety include acetyl, tert-butoxycarbonyl, and benzyloxycarbonyl. Exemplary alkylaminos include NH(C1-C 10 Alkyl(N(C1-C))(N(C1-C)))))))))))))))), example, dialkylamino(C1-C))))))), including, but not limited to these, 10 This includes, but is not limited to, alkyl)2, such as N(CH3)2, -N(CH2CH3)2, -N(CH2CH2CH3)2, and -N(CH(CH3)2)2.

[0497] The term "aminoalkyl" refers to alkyl, alkenyl, and alkynyl compounds as defined above, except that one or more substituted or unsubstituted nitrogen atoms (-N-) are located at the carbon atoms of the alkyl, alkenyl, or alkynyl group. For example, (C2-C6)aminoalkyl refers to a chain containing 2 to 6 carbon atoms and one or more nitrogen atoms located between carbon atoms.

[0498] The terms "hydroxyl" and "hydroxyl" refer to the radical -OH.

[0499] The terms "alkoxyl" or "alkoxy" as used herein refer to an alkyl group to which an oxygen radical is bonded, as defined above, and may be represented by one of -O-alkyl, -O-alkenyl, and -O-alkynyl groups. Aloxy groups may be represented by -O-aryl or O-heteroaryl groups, where aryl and heteroaryl are as defined herein. Alkoxy and alkoxy groups may be substituted with alkyl groups as described above. Exemplary alkoxy groups include, but are not limited to, O-methyl, O-ethyl, O-propyl, O-isopropyl, O-butyl, O-isobutyl, O-sec-butyl, O-tert-butyl, O-pentyl, O-hexyl, O-cyclopropyl, O-cyclobutyl, O-cyclopentyl, and O-cyclohexyl.

[0500] The term "carbonyl" used here refers to the radical -C(O)-. It should be noted that the carbonyl group can be further substituted with various substituents to form other carbonyl groups, including acids, acid halides, amides, esters, and ketones.

[0501] The term "oxo" used here refers to oxygen with a double bond, i.e., =O.

[0502] The term "carboxy" refers to the radical -C(O)O-. It should be noted that the compounds described herein that contain a carboxyl moiety may include their protected derivatives, i.e., those in which the oxygen atom is substituted with a protecting group. Suitable protecting groups for the carboxyl moiety include benzyl and tert-butyl. The carboxyl group used herein includes the -COOH group, i.e., the carboxyl group.

[0503] The term "ester" refers to a chemical part having the formula -C(=O)OR (wherein R is selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl, and heterocycloalkyl).

[0504] The term "cyano" refers to the radical -CN.

[0505] The term "nitro" refers to the radical NO2.

[0506] The term "heteroatom" refers to an atom that is not a carbon atom. Specific examples of heteroatoms include, but are not limited to, nitrogen, oxygen, sulfur, and halogens. A "heteroatomic moiety" includes a moiety to which the atom it bonds is not carbon. Examples of heteroatomic moieties include -N=, -NRN-, -N+(O-)=, -O-, -S- or -S(O)2-, -OS(O)2-, and -SS-, where RN is H or a further substituent.

[0507] The terms "alkylthio" and "thioalkoxy" refer to the alkoxy groups defined above, in which an oxygen atom is replaced by sulfur. In preferred embodiments, the "alkylthio" portion is represented by one of the -S-alkyl, -S-alkenyl, and -S-alkynyl groups. Typical alkylthio groups include methylthio and ethylthio. The term "alkylthio" also includes cycloalkyl, alkene and cycloalkene groups, and alkyne groups. "Arylthio" refers to an aryl or heteroaryl group.

[0508] The term "sulfinyl" refers to the radical -SO-. It should be noted that the sulfinyl group can be further substituted with various substituents to form sulfinyl groups including sulfinic acids, sulfinamides, sulfinyl esters, and sulfoxides.

[0509] The term "sulfonyl" refers to the radical -SO2-. It should be noted that the sulfonyl group can be further substituted with various substituents to form a variety of sulfonyl groups, including sulfonic acids (-SO3H), sulfonamides, sulfonate esters, and sulfones.

[0510] The term "thiocarbonyl" refers to the radical -C(S)-. It should be noted that the thiocarbonyl group can be further substituted with various substituents to form a variety of thiocarbonyl groups, including thioacids, thioamides, thioesters, and thioketones.

[0511] "Acyl" refers to an alkyl-CO- group (where alkyl is defined as previously). Exemplary acyl groups include alkyl groups with 1 to approximately 30 carbon atoms. Exemplary acyl groups also include acetyl, propanoyl, 2-methylpropanoyl, butanoyl, and palmitoyl.

[0512] "Aroyl" refers to an aryl-CO- group (where aryl is defined as previously). Exemplary aroyl groups include benzoyl and 1- and 2-naphthoyl.

[0513] "Arylthio" refers to an aryl-S- group (where the aryl group is defined as previously). Exemplary arylthio groups include phenylthio and naphthylthio.

[0514] "Aralkyl" refers to an aryl-alkyl group (where aryl and alkyl are defined above). Exemplary aralkyl groups include benzyl, phenylethyl, and naphthylmethyl.

[0515] "Aralkyloxy" refers to an aralkyl-O- group (where the aralkyl group is defined as previously). An example of an aralkyloxy group is benzyloxy.

[0516] "Aralkylthio" refers to an aralkyl-S- group (where the aralkyl group is defined as previously). An example of an aralkylthio group is benzylthio.

[0517] "Alkoxycarbonyl" refers to an alkyl-O-CO- group. Exemplary alkoxycarbonyl groups include methoxycarbonyl, ethoxycarbonyl, butyloxycarbonyl, and t-butyloxycarbonyl.

[0518] "Aryloxycarbonyl" refers to an aryl-O-CO- group. Exemplary aryloxycarbonyl groups include phenoxy- and naphthoxy-carbonyl groups.

[0519] "Aralkoxycarbonyl" refers to the aralkyl-O-CO- group. An exemplary aralkoxycarbonyl group is benzyloxycarbonyl.

[0520] "Carbamoyl" refers to the H2N-CO- group.

[0521] "Alkylcarbamoyl" refers to the R'RN-CO- group (where one of R and R' is hydrogen, and the other of R and R' is the alkyl group as defined above).

[0522] "Dialkylcarbamoyl" refers to the R'RN-CO- group (where R and R' are each an alkyl group defined earlier, independently).

[0523] "Acyloxy" refers to an acyl-O- group (where acyl is defined as previously). "Acylamino" refers to an acyl-NH- group (where acyl is defined as previously). "Aroylamino" refers to an aroyl-NH- group (where aroyl is defined as previously).

[0524] The term “optionally substituted” means that the specified group or portion is either unsubstituted or substituted with one or more substituents (typically one, two, three, four, five, or six) independently selected from the group of substituents listed below or otherwise specified in the definition of “substituent.” The term “substituent” means a group that “substitutes” a substituent with any atom of the substituent. Suitable substituents include, but are not limited to, halogens, hydroxyl, carboxyl, oxo, nitro, haloalkyl, alkyl, alkenyl, alkynyl, alkaryl, heteroaryl, cyclyl, heterocyclyl, aralkyl, alkoxy, aryloxy, amino, acylamino, alkylcarbanoyl, arylcarbanoyl, aminoalkyl, alkoxycarbonyl, carboxyl, hydroxylalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamide, arenesulfonamide, aralkylsulfonamide, alkylcarbonyl, acyloxy, cyano, or ureido. In some cases, two substituents can form a ring together with the carbon atom to which they are bonded.

[0525] For example, any alkyl, alkenyl, cycloalkyl, heterocyclyl, heteroaryl or aryl may be optionally OH, CN, -SC(O)Ph, oxo(=O), SH, SO2NH2, SO2(C1-C4)alkyl, SO2NH(C1-C4)alkyl, halogen, carbonyl, thiol, cyano, NH2, NH(C1-C4)alkyl, N[(C1-C4)alkyl]2, C(O)NH2, COOH, COOMe, acetyl, (C1-C8)alkyl, O(C1-C8)alkyl, O(C1-C8)haloalkyl, (C2C8)alkenyl, (C2-C8)alkyl Substituted with one, two, three, four or five substituents independently selected from nyl, haloalkyl, thioalkyl, cyanomethylene, alkylaminyl, aryl, heteroaryl, substituted aryl, NH2-C(O)-alkylene, NH(Me)-C(O)-alkylene, CH2-C(O)-alkyl, C(O)-alkyl, alkylcarbonylaminyl, CH2-[CH(OH)]m-(CH2)p-OH, CH2-[CH(OH)]m-(CH2)p-NH2 or CH2-aryl-alkoxy; "m" and "p" are independently 1, 2, 3, 4, 5 or 6.

[0526] In one embodiment, the optionally substituted group is substituted with one substituent. In another embodiment, the optionally substituted group is substituted with two independently selected substituents (which may be the same or different). In yet another embodiment, the optionally substituted group is substituted with three independently selected substituents (which may be the same or different, or any combination of the same and different). In yet another embodiment, the optionally substituted group is substituted with four independently selected substituents (which may be the same or different, or any combination of the same and different). In yet another embodiment, the optionally substituted group is substituted with five independently selected substituents (which may be the same or different, or any combination of the same and different).

[0527] In one embodiment, any alkyl, alkenyl, cycloalkyl, heterocyclyl, heteroaryl or aryl (e.g., alkyl) is optionally a halogen, -OR 22 , -N(R 22 )2, -SR 22 , -C(O)OR 22 ,-C(O)N(R 22 ) is substituted with one, two, or three substituents independently selected from the group consisting of 2, where R 22 is hydrogen or C 1-3 Alkyl (e.g., 2,2,2-trifluoroethyl, 1,3-dimethoxyprop-2-yl). For example, any alkyl, alkenyl, cycloalkyl, heterocyclyl, heteroaryl or aryl (e.g., alkyl) can be halogen, -OR as desired. 22 , -N(R 22 )2, -SR 22 , -C(O)OR 22 ,-C(O)N(R 22 ) is substituted with one or two substituents independently selected from 2, where R 22 is hydrogen or C 1-3 These are alkyl groups (for example, 2,2,2-trifluoroethyl, 1,3-dimethoxyprop-2-yl).

[0528] The "isocyanato" group refers to the NCO group.

[0529] The "thiocyanate" group refers to the CNS group.

[0530] The "isothiocyanate" group is also known as the NCS group.

[0531] "Alcoyloxy" refers to the RC(=O)O- group.

[0532] "Alcoil" refers to the RC(=O)- group.

[0533] It should be understood that the present invention is not limited to the specific methods, protocols, and reagents provided herein, as they may vary. The terms used herein are for the purpose of des...

Claims

1. Formula (I) 【Chemistry 1】 〔Wherein: B is a nucleic acid base optionally modified (e.g., uracil or 5-methyluracil); X is O or S; Each R Ｖ The hydrogen or hydroxyl protecting group (e.g., ethyl or pivaloyloxymethyl ((CH)) is independent of the hydrogen or hydroxyl protecting group ((CH)) ３ ) ３ CC(O)OCH ２ -, POM) and; R ２ and R ３ One of them is hydrogen, halogen, or -OR ２０ And here: R ２０ is hydrogen, a hydroxyl protecting group, optionally substituted C １－６ alkyl (e.g., methyl, 2 - methoxyethyl, 1,3 - dimethoxyprop - 2 - yl, 2-(N - methylamino)-2 - oxoethyl), optionally substituted C ２－６ alkenyl, or optionally substituted C ２－６ alkynyl (e.g., propargyl); R ２ and the other of R ３ is -OR ３０ where: R ３０ This involves binding to hydrogen, a hydroxyl protecting group, a reactive phosphorus group (e.g., a phosphoramidite), a nucleoside or nucleotide, or an oligonucleotide. and salts thereof.

2. The compound has the structure: 【Chemistry 2】 The compound according to Claim 1, or a salt thereof.

3. The compound has the structure: 【Transformation 3】 The compound according to Claim 1, or a salt thereof.

4. Each R Ｖ Hydrogen is produced independently, and each R is produced as desired. Ｖ A compound according to any one of claims 1 to 3, wherein is independently hydrogen.

5. at least one R Ｖ is a hydroxyl protecting group (e.g., ethyl or POM), and optionally each R Ｖ A compound according to any one of claims 1 to 3, wherein is a hydroxyl protecting group.

6. Each R Ｖ ga-CH ２ CH ３ A compound according to any one of claims 1 to 3 or 5.

7. The compound according to any one of Claims 1 to 6, wherein X is O. 【Request Item 8】 【Chemistry 4】 *-P(O)(OH) ２ , *-P(O)(OCH ２ CH ３ ) ２ Or *-P(O)(OCH ２ OC(O)C(CH ３ ) ３ ) ２ A compound selected from the group consisting of, where * represents bonding to the remainder of the compound, according to any one of claims 1 to 6.

9. The compound according to any one of Claims 1 to 7, wherein X is S.

10. The compound has the structure: 【Transformation 5】 The compound according to Claim 1, or a salt thereof.

11. The compound has the structure: 【Transformation 6】 The compound according to Claim 10, or a salt thereof.

12. The compound has the structure: 【Transformation 7】 The compound according to Claim 10, or a salt thereof.

13. The compound according to any one of Claims 1 to 12, wherein B is a modified or protected nucleic acid base.

14. The compound according to Claim 13, wherein B is a protected nucleic acid base containing at least one amine or hydroxyl protecting group.

15. R ２ A compound according to any one of claims 1 to 14, wherein is hydrogen or a halogen (e.g., fluoro).

16. R ２ ga- OR ２０ A compound according to any one of claims 1 to 14.

17. R ２０ C is replaced as desired. １－６ The compound of claim 16, which is alkyl.

18. R ２０ is methyl, ethyl, propyl, 2-methoxyethyl, 1,3-dimethoxyprop-2-yl, or 2-(N-methylamino)-2-oxoethyl, and optionally R ２０ The compound of claim 17, wherein is methyl or 2-(N-methylamino)-2-oxoethyl.

19. R ２０ C is replaced as desired. ２－６ The compound of claim 16, which is an alkenyl.

20. R ２０ C is replaced as desired. ２－６ The compound of claim 16, wherein the compound is an alkynyl (for example, propargyl).

21. R ３ ga- OR ３０ And here, R ３０ A compound according to any one of claims 15 to 20, wherein is a hydrogen atom, a hydroxyl protecting group, or a reactive phosphorus group.

22. R ３０ The compound of claim 21, wherein is a hydrogen or hydroxyl protecting group.

23. R ３０ The compound of claim 22, wherein is hydrogen.

24. R ３０ The compound of claim 22, wherein is a hydroxyl protecting group.

25. The hydroxyl protecting group is t-butyldimethylsilyl (TBDMS), trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethyltexylsilyl, t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), or t-butylmethoxyphenylsilyl (TBMP), optionally, the hydroxyl protecting group is TBDMS. The compound according to Claim 24.

26. R ３０ The compound of claim 21, wherein is a reactive phosphorus group.

27. The reactive phosphorus group is phosphoramidite, H-phosphonate, alkyl-phosphonate, or phosphate triester. The compound according to Claim 26.

28. Reactive phosphorus group -P(OR) Ｐ )N(R Ｐ２ ) ２ , -P(SR Ｐ )N(R Ｐ２ ) ２ , -P(O)(OR Ｐ )N(R Ｐ２ ) ２ , -P(S)(OR Ｐ )N(R Ｐ２ ) ２ , -P(R Ｐ３ )N(R Ｐ２ ) ２ , -P(O)(SR Ｐ )N(R Ｐ２ ) ２ , -P(O)(OR Ｐ )H, -P(S)(OR Ｐ )H, -P(O)(SR Ｐ )H, -P(O)(OR Ｐ )R Ｐ３ , -P(S)(OR Ｐ )R Ｐ３ , or -P(O)(SR Ｐ )R Ｐ３ And here: Each R Ｐ３ C is replaced as desired. １ -C ３０ Alkyl, optionally substituted C ２ -C ３０ alkenyl, or C substituted as desired. ２ -C ３０ Alkinyl (for example, C which is substituted as desired) １ -C １０ Alkyl, optionally substituted C ２ -C １０ alkenyl, or C substituted as desired. ２ -C １０ Alkinyl) is; Each R Ｐ is independently optionally substituted C １－６ alkyl; and Each R Ｐ２ C is independently replaced as desired. １－６ Is it alkyl? or both R Ｐ２ These, together with the nitrogen atoms to which they are bonded, form 3- to 8-membered heterocyclines which are optionally substituted; or R Ｐ and R Ｐ２ One of them forms a 4- to 8-membered heterocycline, which is substituted as desired with the atom to which they are bonded. The compound according to Claim 27.

29. The reactive phosphorus group is -P(OR Ｐ )N(R Ｐ２ ) ２ and is the compound of claim 28.

30. R Ｐ C is substituted with cyano or -SC(O)Ph １－６ The compound of claim 28 or 29, which is alkyl.

31. R Ｐ ga-CH ２ CH ２ A compound according to any one of claims 28 to 30, which is a CN.

32. Each R Ｐ２ A compound according to any one of claims 28 to 31, wherein is independently methyl, ethyl, propyl, or isopropyl.

33. Each R Ｐ２ A compound according to any one of claims 28 to 32, wherein isopropyl.

34. R Ｐ３ C is replaced as desired. １ -C ６ A compound according to any one of claims 28 to 33, wherein the compound is alkyl (for example, methyl).

35. R ３ ga- OR ３０ And here, R ３０ A compound according to any one of claims 15 to 20, wherein the bond is to a nucleoside or nucleotide, or to an oligonucleotide.

36. R ３０ The compound of claim 35, wherein the bond is to an oligonucleotide.

37. R ３ The compound of claim 36, wherein the compound is bound to an oligonucleotide via a phosphodiester or modified nucleotide bond (e.g., phosphorothioate).

38. R ３ The compound of claim 36 or 37, wherein the compound is bonded to the 5' end of the oligonucleotide (for example, the 5'-hydroxyl group at the 5' end of the oligonucleotide).

39. R ３ A compound according to any one of claims 1 to 14, wherein is hydrogen or a halogen (e.g., F).

40. R ３ ga- OR ２０ A compound according to any one of claims 1 to 14.

41. R ２０ C is replaced as desired. １－６ The compound of claim 40, which is alkyl.

42. R ２０ is methyl, ethyl, propyl, 2-methoxyethyl, 1,3-dimethoxyprop-2-yl, or 2-(N-methylamino)-2-oxoethyl 2-methoxyethyl, 1,3-dimethoxyprop-2-yl, or 2-(N-methylamino)-2-oxoethyl, and optionally R ２０ The compound of claim 41, wherein is methyl or 2-(N-methylamino)-2-oxoethyl.

43. R ２０ C is replaced as desired. ２－６ The compound of claim 40, which is an alkenyl.

44. R ２０ C is replaced as desired. ２－６ The compound of claim 40, wherein the compound is an alkenyl (e.g., propargyl).

45. R ２ ga- OR ３０ And here, R ３０ A compound according to any one of claims 39 to 44, wherein is a hydrogen atom, a hydroxyl protecting group, or a reactive phosphorus group.

46. R ３０ The compound of claim 45, wherein is a hydrogen or hydroxyl protecting group.

47. R ３０ The compound of claim 46, wherein is hydrogen.

48. R ３０ The compound of claim 47, wherein is a hydroxyl protecting group.

49. The compound of claim 48, wherein the hydroxyl protecting group is t-butyldimethylsilyl (TBDMS), trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethyltexylsilyl, t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), or t-butylmethoxyphenylsilyl (TBMPS), preferably the hydroxyl protecting group is TBDMS.

50. R ３０ The compound of claim 45, wherein is a reactive phosphorus group.

51. The compound of claim 50, wherein the reactive phosphorus group is a phosphoramidite, an H-phosphonate, an alkyl-phosphonate, or a phosphoric acid triester.

52. Reactive phosphorus group -P(OR) Ｐ )N(R Ｐ２ ) ２ , -P(SR Ｐ )N(R Ｐ２ ) ２ , -P(O)(OR Ｐ )N(R Ｐ２ ) ２ , -P(S)(OR Ｐ )N(R Ｐ２ ) ２ , -P(R Ｐ３ )N(R Ｐ２ ) ２ , -P(O)(SR Ｐ )N(R Ｐ２ ) ２ , -P(O)(OR Ｐ )H, -P(S)(OR Ｐ )H, -P(O)(SR Ｐ )H, -P(O)(OR Ｐ )R Ｐ３ , -P(S)(OR Ｐ )R Ｐ３ , or -P(O)(SR Ｐ )R Ｐ３ And here: Each R Ｐ３ C is replaced as desired. １ -C ３０ Alkyl, optionally substituted C ２ -C ３０ alkenyl, or C substituted as desired. ２ -C ３０ Alkinyl (for example, C which is substituted as desired) １ -C １０ Alkyl, optionally substituted C ２ -C １０ alkenyl, or C substituted as desired. ２ -C １０ Alkinyl) is; Each R Ｐ C is independently replaced as desired. １－６ It is alkyl; and Each R Ｐ２ C is independently replaced as desired. １－６ Is it alkyl? or both R Ｐ２ These, together with the nitrogen atoms to which they are bonded, form 3- to 8-membered heterocyclines which are optionally substituted; or R Ｐ and R Ｐ２ One of them forms a 4- to 8-membered heterocycline, which is substituted as desired with the atom to which they are bonded. The compound according to claim 51.

53. Reactive phosphorus group -P(OR) Ｐ )N(R Ｐ２ ) ２ The compound according to claim 52.

54. R Ｐ C is substituted with cyano or -SC(O)Ph １－６ The compound of claim 52 or 53, which is alkyl.

55. R Ｐ ga-CH ２ CH ２ A compound according to any one of claims 52 to 54, which is a CN.

56. Each R Ｐ２ A compound according to any one of claims 52 to 55, wherein is independently methyl, ethyl, propyl, or isopropyl.

57. Each R Ｐ２ A compound according to any one of claims 52 to 56, wherein isopropyl.

58. R Ｐ３ C is replaced as desired. １ -C ６ A compound according to any one of claims 52 to 57, wherein the compound is alkyl (for example, methyl).

59. R ２ ga- OR ３０ And here, R ３０ A compound according to any one of claims 39 to 44, wherein the bond is to a nucleoside or nucleotide, or to an oligonucleotide.

60. R ３０ The compound of claim 59, wherein the bond is to an oligonucleotide.

61. R ２ The compound of claim 60, wherein the compound is bound to an oligonucleotide via a phosphodiester or modified nucleotide bond (e.g., phosphorothioate).

62. R ２ The compound of claim 60 or 61, wherein the compound is bonded to the 5' end of the oligonucleotide (for example, the 5'-hydroxyl group at the 5' end of the oligonucleotide).

63. R ２ is hydrogen, F, or -OR ２０ And, Here, R ２０ C is substituted with hydrogen, if desired. １－６ Alkyl (e.g., methyl, 2-methoxyethyl, 1,3-dimethoxyprop-2-yl, 2-(N-methylamino)-2-oxoethyl), optionally substituted C ２－６ alkenyl, or C substituted as desired. ２－６ Alkinyl (e.g., propargyl); and R ３ ga- OR ３０ And, Here, R ３０ is a hydrogen atom, a hydroxyl protecting group, or a reactive phosphorus group. A compound according to any one of claims 1 to 6.

64. R ３０ The compound of claim 63, wherein is a hydrogen or hydroxyl protecting group.

65. R ３０ The compound of claim 63, wherein is a reactive phosphorus group.

66. The compound of claim 65, wherein the reactive phosphorus group is a phosphoramidite, an H-phosphonate, an alkyl-phosphonate, or a phosphoric acid triester.

67. The reactive phosphorus group is -P(OR Ｐ１ )N(R Ｐ２ ) ２ , -P(SR Ｐ１ )N(R Ｐ２ ) ２ , -P(O)(OR Ｐ１ )N(R Ｐ２ ) ２ , -P(S)(OR Ｐ１ )N(R Ｐ２ ) ２ , -P(R Ｐ３ )N(R Ｐ２ ) ２ , -P(O)(SR Ｐ１ )N(R Ｐ２ ) ２ , -P(O)(OR Ｐ１ )H, -P(S)(OR Ｐ１ )H, -P(O)(SR Ｐ１ )H, -P(O)(OR Ｐ１ )R Ｐ３ , -P(S)(OR Ｐ１ )R Ｐ３ , or -P(O)(SR Ｐ１ )R Ｐ３ And here: Each R Ｐ３ C is replaced as desired. １ -C ３０ Alkyl, optionally substituted C ２ -C ３０ alkenyl, or C substituted as desired. ２ -C ３０ Alkinyl (for example, C which is substituted as desired) １ -C １０ Alkyl, optionally substituted C ２ -C １０ alkenyl, or C substituted as desired. ２ -C １０ Alkinyl) is; Each R Ｐ１ C is independently replaced as desired. １－６ It is alkyl; and Each R Ｐ２ C is independently replaced as desired. １－６ Is it alkyl? or both R Ｐ２ These, together with the nitrogen atoms to which they are bonded, form 3- to 8-membered heterocyclines which are optionally substituted; or R Ｐ１ and R Ｐ２ One of them forms a 4- to 8-membered heterocycline, which is substituted as desired with the atom to which they are bonded. The compound according to claim 66.

68. Reactive phosphorus group -P(OR) Ｐ１ )N(R Ｐ２ ) ２ The compound according to claim 67.

69. R Ｐ１ C is substituted with cyano or -SC(O)Ph １－６ The compound of claim 67 or 68, which is alkyl.

70. R Ｐ１ ga-CH ２ CH ２ A compound according to any one of claims 67 to 69, which is CN.

71. Each R Ｐ２ A compound according to any one of claims 67 to 70, wherein is independently methyl, ethyl, propyl, or isopropyl.

72. Each R Ｐ２ A compound according to any one of claims 67 to 71, wherein isopropyl.

73. Reactive phosphorus group -P(OR) Ｐ１ )N(R Ｐ２ ) ２ And here, R Ｐ１ ga-CH ２ CH ２ CN, and each R Ｐ２ A compound according to any one of claims 66 to 72, wherein isopropyl.

74. R ２ is hydrogen, F, or -OR ２０ And here, R ２０ A compound according to any one of claims 63 to 73, wherein is methyl or 2-methoxyethyl.

75. Each R Ｖ is independently hydrogen (for example, 【Transformation 8】 *-P(O)(OH) ２ A compound according to any one of claims 63 to 74.

76. Each R Ｖ These are independently hydroxyl protecting groups (for example, 【Chemistry 9】 *-P(O)(OCH ２ CH ３ ) ２ Or *-P(O)(OCH ２ OC(O)C(CH ３ ) ３ ) ２ A compound according to any one of claims 64 to 74.

77. The compound is: 【Chemistry 10】 A compound according to claim 1, selected from the group consisting of the following.

78. structure: 【Chemistry 11】 [In the ceremony: X is either O or S; and Each R Ｖ These are independently hydrogen or hydroxyl protecting groups. An oligonucleotide having a 5'-terminus modification including the following.

79. 5'-terminal modification is structural: 【Chemistry 12】 The oligonucleotide of claim 78, comprising

80. 5'-terminal modification is structural: 【Chemistry 13】 The oligonucleotide of claim 78, comprising

81. The 5' terminal nucleotide has structure: 【Chemistry 14】 [In the ceremony: B is a nucleic acid base that is modified as desired (e.g., uracil); Each R Ｖ The hydrogen or hydroxyl protecting group (e.g., ethyl or pivaloyloxymethyl ((CH)) is independently determined. ３ ) ３ CC(O)OCH ２ -, POM) and; R ２ and R ３ One of them is hydrogen, halogen, or -OR ２０ And here: R ２０ C is substituted with hydrogen, a hydroxyl protecting group, or optionally C １－６ Alkyl (e.g., methyl, 2-methoxyethyl, 1,3-dimethoxyprop-2-yl, 2-(N-methylamino)-2-oxoethyl), optionally substituted C ２－６ alkenyl, or C substituted as desired. ２－６ It is alkinyl (for example, propargyl); R ２ and R ３ The other is - OR ３０ And here: R ３０ This is binding to an oligonucleotide (for example, an internucleotide bond that connects to a subsequent nucleotide of the oligonucleotide). Oligonucleotides having the properties of an oligonucleotide.

82. The 5' terminal nucleotide has structure: 【Chemistry 15】 The oligonucleotide of claim 81, having the properties of the present invention.

83. The compound's structure: 【Chemistry 16】 The compound according to claim 81.

84. The 5' terminal nucleotide has structure: 【Chemistry 17】 The oligonucleotide of claim 81, having the properties of the present invention.

85. The 5' terminal nucleotide has structure: [Chemistry 18] The oligonucleotide of claim 84, having the properties of the oligonucleotide of claim 84.

86. The 5' terminal nucleotide has structure: 【Chemistry 19】 The oligonucleotide of claim 84, having the properties of the oligonucleotide of claim 84.

87. R ３ ga- OR ３０ The oligonucleotide according to any one of claims 79 to 86.

88. An oligonucleotide wherein the oligonucleotide is a compound of any of claims 35 to 38 or 59 to 62.

89. An oligonucleotide according to any one of claims 78 to 88, wherein the oligonucleotide is 10 to 50 nucleotides long (for example, 15 to 40 nucleotides), and the compound of formula (I) is one nucleotide.

90. The oligonucleotide of claim 89, wherein the oligonucleotide is 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides long, and optionally, the oligonucleotide is 17, 18, 19, 21, 22, 23, 24, or 25 nucleotides long.

91. An oligonucleotide according to any one of claims 78 to 90, wherein the oligonucleotide comprises at least one nucleic acid modification (e.g., one, two, three, four, five, six, seven, eight, nine, ten or more independently selected modifications).

92. The oligonucleotide according to claim 91, wherein the oligonucleotide comprises at least one nucleic acid modification selected from the group consisting of nucleic acid base modification, sugar modification, internucleotide bond modification, conjugate (e.g., ligand), and any combination thereof.

93. An oligonucleotide according to any one of claims 78 to 92, wherein the oligonucleotide comprises at least one (for example, one, two, three, four, five, six, seven, eight, nine, ten or more) 2'-OMe nucleotides.

94. An oligonucleotide according to any one of claims 78 to 93, wherein the oligonucleotide comprises at least one (e.g., one, two, three, four, five, six, seven, eight, nine, ten or more) double-stranded thermal destabilization modifications.

95. The oligonucleotide of claim 94, wherein the double-chain thermal destabilization modification is located at the 4th, 5th, 6th, 7th, or 8th position counting from the 5' end of the oligonucleotide, where the compound of formula (I) is at the 1st position from the 5' end of the oligonucleotide, and optionally the double-chain thermal destabilization modification is located at the 6th, 7th, or 8th position counting from the 5' end of the oligonucleotide, preferably the double-chain thermal destabilization modification is located at the 7th position counting from the 5' end of the oligonucleotide.

96. An oligonucleotide according to any one of claims 78 to 95, wherein the oligonucleotide comprises at least one (for example, one, two, three, four, five, six, seven, eight, nine, ten or more) 2'-F nucleotides.

97. The oligonucleotide of claim 96, wherein the oligonucleotide contains a 2'-F nucleotide at at least the 2nd, 14th, and 16th positions, counting from the 5' end of the oligonucleotide, where the compound of formula (I) is at position 1 from the 5' end of the oligonucleotide, and optionally the oligonucleotide contains a 2'-F nucleotide at at least the 2nd, 6th, 14th, and 16th positions, counting from the 5' end of the oligonucleotide, preferably the oligonucleotide contains a 2'-F nucleotide at at least the 2nd, 6th, 9th, 14th, and 16th positions, counting from the 5' end of the oligonucleotide, or at least the 2nd, 6th, 8th, 9th, 14th, and 16th positions.

98. A compound according to any one of claims 78 to 97, wherein the oligonucleotide comprises at least one (for example, one, two, three, four, five, six, seven, eight, nine, ten or more) 2'-deoxy(2'-H) nucleotides.

99. The oligonucleotide of claim 98, wherein the oligonucleotide comprises a 2'-deoxynucleotide at any one of the following positions, counting from the 5' end of the oligonucleotide: position 2, position 5, position 7, position 12, position 14, and position 16, and the compound of formula (I) is at position 1 from the 5' end of the oligonucleotide, and optionally the oligonucleotide comprises a 2'-deoxynucleotide at at least position 5, counting from the 5' end of the oligonucleotide, preferably the oligonucleotide comprises a 2'-deoxynucleotide at at least positions 2, 5, and 9, or at least positions 2, 5, 7, and 12, or at least positions 2, 5, 7, 12, 14, and 16, counting from the 5' end of the oligonucleotide.

100. An oligonucleotide according to any one of claims 78 to 99, wherein the oligonucleotide comprises at least one (for example, one, two, three, four, five, six, seven, eight, nine, ten or more) unnatural or modified nucleic acid bases.

101. An oligonucleotide according to any one of claims 78 to 100, wherein the oligonucleotide contains at least one (for example, one, two, three, four, five, six, seven, eight, nine, ten or more) modified nucleoside bonds.

102. The oligonucleotide according to claims 78 to 101, wherein the oligonucleotide contains phosphorothioate bonds between the nucleotides at positions 1 and 2, and between the nucleotides at positions 2 and 3, counting from the 5' end of the oligonucleotide, where the compound of formula (I) is the nucleotide at position 1 from the 5' end of the oligonucleotide; and the oligonucleotide contains phosphorothioate bonds between the nucleotides at positions 1 and 2, and between the nucleotides at positions 2 and 3, counting from the 3' end of the oligonucleotide.

103. An oligonucleotide according to any one of claims 78 to 102, wherein the oligonucleotide is covalently bonded to a support, for example, a solid support.

104. A double-stranded RNA (dsRNA) comprising a sense strand and an antisense strand, wherein the sense strand is substantially complementary to the antisense strand, and where either the sense strand or the antisense strand is an oligonucleotide of any of claims 78 to 103.

105. The dsRNA of claim 104, wherein the antisense strand is an oligonucleotide of any of claims 78 to 103.

106. The dsRNA according to claim 104 or 105, wherein the dsRNA can induce RNA interference.

107. A method for reducing the expression of a target gene in a subject, comprising administering to the subject (i) any double-stranded RNA according to claims 104 to 106 (where the antisense strand is substantially complementary to the target gene); or (ii) any oligonucleotide according to claims 78 to 103 (where the oligonucleotide is substantially complementary to the target gene).

108. A composition comprising any compound of claims 1 to 77, any oligonucleotide of claims 78 to 103, or any dsRNA of claims 104 to 106.

109. A kit comprising any compound of claims 1 to 77, any oligonucleotide of claims 78 to 103, or any dsRNA of claims 104 to 106.

110. A cell comprising any compound of claims 1 to 77, any oligonucleotide of claims 78 to 103, or any dsRNA of claims 104 to 106.

111. The cell according to claim 110, wherein the cell is in vivo.

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